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Sample records for characterize degradation pathways

  1. Cathodic degradation of antibiotics: characterization and pathway analysis.

    Science.gov (United States)

    Kong, Deyong; Liang, Bin; Yun, Hui; Cheng, Haoyi; Ma, Jincai; Cui, Minhua; Wang, Aijie; Ren, Nanqi

    2015-04-01

    Antibiotics in wastewaters must be degraded to eliminate their antibacterial activity before discharging into the environment. A cathode can provide continuous electrons for the degradation of refractory pollutants, however the cathodic degradation feasibility, efficiency and pathway for different kinds of antibiotics is poorly understood. Here, we investigated the degradation of four antibiotics, namely nitrofurazone (NFZ), metronidazole (MNZ), chloramphenicol (CAP), and florfenicol (FLO) by a poised cathode in a dual chamber electrochemical reactor. The cyclic voltammetry preliminarily proved the feasibility of the cathodic degradation of these antibiotics. The cathodic reducibility of these antibiotics followed the order of NFZ > MNZ > CAP > FLO. A decreased phosphate buffered solution (PBS) concentration as low as 2 mM or utilization of NaCl buffer solution as catholyte had significant influence on antibiotics degradation rate and efficiency for CAP and FLO but not for NFZ and MNZ. PBS could be replaced by Na2CO3-NaHCO3 buffer solution as catholyte for the degradation of these antibiotics. Reductive dechlorination of CAP proceeded only after the reduction of the nitro group to aromatic amine. The composition of the degradation products depended on the cathode potential except for MNZ. The cathodic degradation process could eliminate the antibacterial activity of these antibiotics. The current study suggests that the electrochemical reduction could serve as a potential pretreatment or advanced treatment unit for the treatment of antibiotics containing wastewaters. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Identification and characterization of process-related substances and degradation products in apremilast: Process optimization and degradation pathway elucidation.

    Science.gov (United States)

    Lu, Yuting; Shen, Xiaoyue; Hang, Taijun; Song, Min

    2017-07-15

    This study aims at investigating the separation, identification and characterization of related substances in apremilast by LC-MS hyphenated techniques, as well as the synthesis optimization and the degradation pathways elucidation. Forced degradation studies were conducted under the ICH prescribed stress conditions. The chromatographic separation was achieved on XBridge C18 column (4.6mm×150mm, 3.5μm) using a mobile phase consisting of water adjusted to pH 3.0 with formic acid as solvent A and acetonitrile as solvent B in linear gradient elution program. Twelve related substances were detected all together in apremilast and its stress samples. Their structures were identified mainly through positive ESI high-resolution TOF-MS analysis of the parent ions' accurate masses and elemental compositions, and the corresponding MS/MS spectra elucidation. There were three process-related substances and nine degradation products, seven of them were first reported. Two degradation products and one process-related substance were further verified by semi-preparation and NMR determination. Their origins and formation mechanisms were also discussed, based on which effective approaches for the synthesis optimization were conducted. Therefore, the related substances investigation are valuable for apremilast manufacturing process optimization and quality control. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Proteogenomic Characterization of Monocyclic Aromatic Hydrocarbon Degradation Pathways in the Aniline-Degrading Bacterium Burkholderia sp. K24.

    Directory of Open Access Journals (Sweden)

    Sang-Yeop Lee

    Full Text Available Burkholderia sp. K24, formerly known as Acinetobacter lwoffii K24, is a soil bacterium capable of utilizing aniline as its sole carbon and nitrogen source. Genomic sequence analysis revealed that this bacterium possesses putative gene clusters for biodegradation of various monocyclic aromatic hydrocarbons (MAHs, including benzene, toluene, and xylene (BTX, as well as aniline. We verified the proposed MAH biodegradation pathways by dioxygenase activity assays, RT-PCR, and LC/MS-based quantitative proteomic analyses. This proteogenomic approach revealed four independent degradation pathways, all converging into the citric acid cycle. Aniline and p-hydroxybenzoate degradation pathways converged into the β-ketoadipate pathway. Benzoate and toluene were degraded through the benzoyl-CoA degradation pathway. The xylene isomers, i.e., o-, m-, and p-xylene, were degraded via the extradiol cleavage pathways. Salicylate was degraded through the gentisate degradation pathway. Our results show that Burkholderia sp. K24 possesses versatile biodegradation pathways, which may be employed for efficient bioremediation of aniline and BTX.

  4. Characterization of the novel dimethyl sulfide-degrading bacterium Alcaligenes sp. SY1 and its biochemical degradation pathway

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Yiming; Qiu, Jiguo; Chen, Dongzhi; Ye, Jiexu; Chen, Jianmeng, E-mail: jchen@zjut.edu.cn

    2016-03-05

    Highlights: • A novel efficient DMS-degrading bacterium Alcaligenes sp. SY1 was identified. • A RSM was applied to optimize incubation condition of Alcaligenes sp. SY1. • SIP was applied as C{sup 13} labelled DMS to trace intermediates during DMS degradation. • Kinetics of DMS degradation via batch experiment was revealed. • Carbon and sulfur balance were analyzed during DMS degradation process. - Abstract: Recently, the biodegradation of volatile organic sulfur compounds (VOSCs) has become a burgeoning field, with a growing focus on the reduction of VOSCs. The reduction of VOSCs encompasses both organic emission control and odor control. Herein, Alcaligenes sp. SY1 was isolated from active sludge and found to utilize dimethyl sulfide (DMS) as a growth substrate in a mineral salt medium. Response surface methodology (RSM) analysis was applied to optimize the incubation conditions. The following conditions for optimal degradation were identified: temperature 27.03 °C; pH 7.80; inoculum salinity 0.84%; and initial DMS concentration 1585.39 μM. Under these conditions, approximately 99% of the DMS was degraded within 30 h of incubation. Two metabolic compounds were detected and identified by gas chromatography–mass spectrometry (GC–MS): dimethyl disulfide (DMDS) and dimethyl trisulfide (DMTS). The DMS degradation kinetics for different concentrations were evaluated using the Haldane–Andrews model and the pseudo first-order model. The maximum specific growth rate and degradation rate of Alcaligenes sp. SY1 were 0.17 h{sup −1} and 0.63 gs gx{sup −1} h{sup −1}. A possible degradation pathway is proposed, and the results suggest that Alcaligenes sp. SY1 has the potential to control odor emissions under aerobic conditions.

  5. Characterization of the Complete Uric Acid Degradation Pathway in the Fungal Pathogen Cryptococcus neoformans

    Science.gov (United States)

    Lee, I. Russel; Yang, Liting; Sebetso, Gaseene; Allen, Rebecca; Doan, Thi H. N.; Blundell, Ross; Lui, Edmund Y. L.; Morrow, Carl A.; Fraser, James A.

    2013-01-01

    Degradation of purines to uric acid is generally conserved among organisms, however, the end product of uric acid degradation varies from species to species depending on the presence of active catabolic enzymes. In humans, most higher primates and birds, the urate oxidase gene is non-functional and hence uric acid is not further broken down. Uric acid in human blood plasma serves as an antioxidant and an immune enhancer; conversely, excessive amounts cause the common affliction gout. In contrast, uric acid is completely degraded to ammonia in most fungi. Currently, relatively little is known about uric acid catabolism in the fungal pathogen Cryptococcus neoformans even though this yeast is commonly isolated from uric acid-rich pigeon guano. In addition, uric acid utilization enhances the production of the cryptococcal virulence factors capsule and urease, and may potentially modulate the host immune response during infection. Based on these important observations, we employed both Agrobacterium-mediated insertional mutagenesis and bioinformatics to predict all the uric acid catabolic enzyme-encoding genes in the H99 genome. The candidate C. neoformans uric acid catabolic genes identified were named: URO1 (urate oxidase), URO2 (HIU hydrolase), URO3 (OHCU decarboxylase), DAL1 (allantoinase), DAL2,3,3 (allantoicase-ureidoglycolate hydrolase fusion protein), and URE1 (urease). All six ORFs were then deleted via homologous recombination; assaying of the deletion mutants' ability to assimilate uric acid and its pathway intermediates as the sole nitrogen source validated their enzymatic functions. While Uro1, Uro2, Uro3, Dal1 and Dal2,3,3 were demonstrated to be dispensable for virulence, the significance of using a modified animal model system of cryptococcosis for improved mimicking of human pathogenicity is discussed. PMID:23667704

  6. Biochemical and structural characterization of Klebsiella pneumoniae oxamate amidohydrolase in the uric acid degradation pathway

    Energy Technology Data Exchange (ETDEWEB)

    Hicks, Katherine A.; Ealick, Steven E.

    2016-05-25

    HpxW from the ubiquitous pathogenKlebsiella pneumoniaeis involved in a novel uric acid degradation pathway downstream from the formation of oxalurate. Specifically, HpxW is an oxamate amidohydrolase which catalyzes the conversion of oxamate to oxalate and is a member of the Ntn-hydrolase superfamily. HpxW is autoprocessed from an inactive precursor to form a heterodimer, resulting in a 35.5 kDa α subunit and a 20 kDa β subunit. Here, the structure of HpxW is presented and the substrate complex is modeled. In addition, the steady-state kinetics of this enzyme and two active-site variants were characterized. These structural and biochemical studies provide further insight into this class of enzymes and allow a mechanism for catalysis consistent with other members of the Ntn-hydrolase superfamily to be proposed.

  7. Molecular characterization of the Akt-TOR signaling pathway in rainbow trout: potential role in muscle growth/degradation

    Science.gov (United States)

    The Akt-TOR signaling pathway plays a key role in cellular metabolism and muscle growth. Hormone, nutrition and stress factors affect the Akt-TOR pathway by regulating gene transcription, protein synthesis and degradation. In addition, we previously showed that energetic demands elevate during vit...

  8. Characterization of a novel β-cypermethrin-degrading Aspergillus niger YAT strain and the biochemical degradation pathway of β-cypermethrin.

    Science.gov (United States)

    Deng, Weiqin; Lin, Derong; Yao, Kai; Yuan, Huaiyu; Wang, Zhilong; Li, Jianlong; Zou, Likou; Han, Xinfeng; Zhou, Kang; He, Li; Hu, Xinjie; Liu, Shuliang

    2015-10-01

    Aspergillus niger YAT strain was obtained from Chinese brick tea (Collection number: CGMCC 10,568) and identified on the basis of morphological characteristics and internal transcribed spacer (ITS) sequence. The strain could degrade 54.83 % of β-cypermethrin (β-CY; 50 mg L(-1)) in 7 days and 100 % of 3-phenoxybenzoic acid (3-PBA; 100 mg L(-1)) in 22 h. The half-lives of β-CY and 3-PBA range from 3.573 to 11.748 days and from 5.635 to 12.160 h, respectively. The degradation of β-CY and 3-PBA was further described using first-order kinetic models. The pathway and mechanism of β-CY degraded by YAT were investigated by analyzing the degraded metabolites through high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). Relevant enzymatic activities and substrate utilization were also investigated. β-CY degradation products were analyzed. Results indicated that YAT strain transformed β-CY into 3-PBA. 3-PBA was then gradually transformed into permethric acid, protocatechuic acid, 3-hydroxy-5-phenoxy benzoic acid, gallic acid, and phenol gradually. The YAT strain can also effectively degrade these metabolites. The results indicated that YAT strain has potential applications in bioremediation of pyrethroid insecticide (PI)-contaminated environments and fermented food.

  9. Identification and characterization of the furfural and 5-(hydroxymethyl)furfural degradation pathways of Cupriavidus basilensis HMF14

    OpenAIRE

    Koopman, Frank; Wierckx, Nick; de Winde, Johannes H; Ruijssenaars, Harald J.

    2010-01-01

    The toxic fermentation inhibitors in lignocellulosic hydrolysates pose significant problems for the production of second-generation biofuels and biochemicals. Among these inhibitors, 5-(hydroxymethyl)furfural (HMF) and furfural are specifically notorious. In this study, we describe the complete molecular identification and characterization of the pathway by which Cupriavidus basilensis HMF14 metabolizes HMF and furfural. The identification of this pathway enabled the construction of an HMF an...

  10. Vacuole import and degradation pathway:Insights into a specialized autophagy pathway

    Institute of Scientific and Technical Information of China (English)

    Abbas; A; Alibhoy; Hui-Ling; Chiang

    2011-01-01

    Glucose deprivation induces the synthesis of pivotagluconeogenic enzymes such as fructose-1,6-bisphos-phatase, malate dehydrogenase, phosphoenolpyruvatecarboxykinase and isocitrate lyase in Saccharomycescerevisiae. However, following glucose replenishment,these gluconeogenic enzymes are inactivated and de-graded. Studies have characterized the mechanismsby which these enzymes are inactivated in response toglucose. The site of degradation of these proteins hasalso been ascertained to be dependent on the dura-tion of starvation. Glucose replenishment of short-termstarved cells results in these proteins being degradedin the proteasome. In contrast, addition of glucose tocells starved for a prolonged period results in theseproteins being degraded in the vacuole. In the vacuoledependent pathway, these proteins are sequestered inspecialized vesicles termed vacuole import and degra-dation (Vid). These vesicles converge with the endo-cytic pathway and deliver their cargo to the vacuolefor degradation. Recent studies have identified thatinternalization, as mediated by actin polymerization, isessential for delivery of cargo proteins to the vacuolefor degradation. In addition, components of the targetof rapamycin complex 1 interact with cargo proteins during glucose starvation. Furthermore, Tor1p dissoci-ates from cargo proteins following glucose replenish-ment. Future studies will be needed to elaborate on the importance of internalization at the plasma membrane and the subsequent import of cargo proteins into Vid vesicles in the vacuole dependent degradation pathway.

  11. Enzymatic description of the anhydrofructose pathway of glycogen degradation. I

    DEFF Research Database (Denmark)

    Yu, Shukun; Refdahl, Charlotte; Lundt, Inge

    2004-01-01

    The anhydrofructose pathway describes the degradation of glycogen and starch to metabolites via 1,5-anhydro-D-fructose (1,5AnFru). The enzyme catalyzing the first reaction step of this pathway, i.e., a-1,4-glucan lyase (EC 4.2.1.13), has been purified, cloned and characterized from fungi and red...... algae in our laboratory earlier. In the present study, two 1,5AnFru metabolizing enzymes were discovered in the fungus Anthracobia melaloma for the formation of ascopyrone P (APP), a fungal secondary metabolite exhibiting antibacterial and antioxidant activity. These are 1,5AnFru dehydratase (AFDH...

  12. Modeling position-specific isotope fractionation of organic micropollutants degradation via different reaction pathways

    DEFF Research Database (Denmark)

    Jin, Biao; Rolle, Massimo

    : dichlorobenzamide (BAM), isoproturon (IPU) and diclofenac (DCF). The model successfully reproduces the multi-element isotope data, and precisely captures the dual element isotope trends, characterizing the different degradation pathways. Besides illustrating the model capability of mechanistic evaluation...

  13. Microbial PAH-Degradation in Soil: Degradation Pathways and Contributing Factors

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xu-Xiang; CHENG Shu-Pei; ZHU Cheng-Jun; SUN Shi-Lei

    2006-01-01

    Adverse effects on the environment and high persistence in the microbial degradation and environmental fate of polycyclic aromatic hydrocarbons (PAHs) are motivating interest. Many soil microorganisms can degrade PAHs and use various metabolic pathways to do so. However, both the physio-chemical characteristics of compounds as well as the physical, chemical, and biological properties of soils can drastically influence the degradation capacity of naturally occurring microorganisms for field bioremediation. Modern biological techniques have been widely used to promote the efficiency of microbial PAH-degradation and make the biodegradation metabolic pathways more clear. In this review microbial degradation of PAHs in soil is discussed, with emphasis placed on the main degradation pathways and the environmental factors affecting biodegradation.

  14. Inhibitors and pathways of hepatocytic protein degradation.

    Science.gov (United States)

    Seglen, P O; Gordon, P B; Grinde, B; Solheim, A; Kovács, A L; Poli, A

    1981-01-01

    On the basis of experiments using amino acids and various inhibitors (lysosomotropic amines, leupeptin, chymostatin, vanadate, vinblastine, anoxia, methylaminopurines), five different modes of endogenous protein degradation in isolated rat hepatocytes can be distinguished. The two non-lysosomal (amine-resistant) mechanisms preferentially degrade relatively labile (short-lived) proteins: one of these mechanisms is energy-dependent and chymostatin-sensitive, the other is not. Of the three lysosomal (amine-sensitive) mechanisms, one--quantitatively minor--is amino acid-resistant and preferentially degrades labile proteins. The two amino acid-sensitive mechanisms each seen account for about one-half of the degradation of relatively stable (long-lived) proteins; one of them is suppressed by leucine and apparently corresponds to the formation of electron microscopically visible autophagosomes; the other may represent a different type of autophagy, inhibited by asparagine and glutamine. A new class of inhibitors, the purine derivatives (methylated 6-aminopurines, and 6-mercaptopurines) appear to specifically suppress autophagic/lysosomal protein degradation, and may help to further elucidate the mechanisms of autophagy.

  15. Phylogenetic and degradation characterization of Burkholderia cepacia WZ1 degrading herbicide quinclorac.

    Science.gov (United States)

    Lü, Zhenmei; Min, Hang; Wu, Shuwen; Ruan, Aidong

    2003-11-01

    Strain WZI capable of degrading quinclorac was isolated from a pesticide manufactory soil and considered to be Burkholderia cepacia, belonged to bacteria, Proteobacteria, beta-Proteobacteria, based on morphology, physio-biochemical properties, whole cell fatty acid analysis and a partial sequencing of 16S rDNA. Strain WZ1 decomposed 90% of quinclorac at original concentration of 1000 mg L(-1) within 11 days. GC/MS analysis showed that the strain degraded quinclorac to 3,7-dichloro-8-quinoline and the cracked residue 2-chloro, 1,4-benzenedicarboxylic acid, indicating that the metabolic pathway was initiated by process of decarboxylation followed by cleavage of the aromatic ring. Stain WZ1 was also able to degrade some other herbicides and aromatic compounds, including 2,4,5-T, phenol, naphthalene and hydrochinone etc. This paper describes for the first time Phylogenetic and degradation characterization of a pure bacterium which, is able to mineralize quinclorac.

  16. Characterization, Degradation, and Reaction Pathways of Indoor Toluene over Visible-light-driven S, Zn Co-doped TiO2

    Science.gov (United States)

    Chu, H.; Lin, Y. H.; Lin, C. Y.

    2017-01-01

    Sulfur and Zinc co-doped TiO2 prepared by a sol-gel method to degrade toluene under a fluorescent lamp was investigated. The results indicate that S,Zn co-doped TiO2 photocatalysts are mainly nano-size with an anatase phase structure. The degradation reactions of toluene were performed under various operation conditions. The results show that the toluene conversion increases with increasing toluene concentration and decreasing relative humidity. Based on the results of activity test, S0.05Zn0.001/TiO2 was chosen for further studies. The main oxidation products of toluene photodegradation are CO2, H2O, benzyl alcohol, acetone, butadiene and acetic acid. Two possible mechanisms have been developed for photodegradation of toluene in a dry and a humid environment.

  17. Phenanthrene-degrading pathway of Agrobacterium sp. Phx1

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Lei; YUAN; Hongli; WANG; Shuangqing; HUANG; Huaize

    2005-01-01

    The metabolic pathway of phenanthrene-degrading strain Agrobacterium sp. Phx1 was investigated. Phx1 almost was able to transform 100 υg/mL of phenanthrene completely in 1 day in broth media of beef extract-peptone (BP), Luria-Bertani (LB) and mineral salts media (MS), and LB and BP could promote the growth and degradation efficiency of Phx1. The GC-MS was employed to analyze the metabolites of the 1st, 3rd, 7th days of phenanthrene degradation in MS. As a result, the 1-Hydroxy-2-naphthoic acid (1H2N) and 1-naphthol (NOL) were detected in the metabolites of the 1st day. Only NOL was observed on the 3rd day and it disappeared on the 7th day. The accumulated NOL did not pertain to the defined pathway of phenanthrene degradation by bacteria. The further HPLC study confirmed the finding in GC-MS analysis and found the production of catechol (CAT) from o-phthalic acid (OPA) in the phenanthrene metabolizing, which has never been reported in the defined degrading pathways. This production was also evidenced by the production of CAT using OPA as substrate. All of our results showed that the Agrobacterium sp. Phx1 had a novel phenanthrene-degrading pathway.

  18. When RNA and protein degradation pathways meet

    Directory of Open Access Journals (Sweden)

    Pascal eGENSCHIK

    2014-04-01

    Full Text Available RNA silencing has become a major focus of molecular and biomedical research in the last decade. This mechanism, which is conserved in most eukaryotes, has been extensively studied and is associated to various pathways implicated in the regulation of development, in the control of transposition events, heterochromatin maintenance and also playing a role in defense against viruses. Despite of its importance, the regulation of the RNA silencing machinery itself remains still poorly explored. Recently several reports in both plants and metazoans revealed that key components of RNA silencing, such as RNA-induced silencing complex (RISC component ARGONAUTE proteins, but also the endonuclease Dicer are subjected to proteasomal and autophagic pathways. Here we will review these post-translational proteolytic regulations with a special emphasis on plant research and also discuss their functional relevance.

  19. Enrichment and characterization of sulfate reducing, naphthalene degrading microorganisms

    Science.gov (United States)

    Steffen, Kümmel; Florian-Alexander, Herbst; Márcia, Duarte; Dietmar, Pieper; Jana, Seifert; Bergen Martin, von; Hans-Hermann, Richnow; Carsten, Vogt

    2014-05-01

    Polycyclic aromatic hydrocarbons (PAH) are pollutants of great concern due to their potential toxicity, mutagenicity and carcinogenicity. PAH are widely distributed in the environment by accidental discharges during the transport, use and disposal of petroleum products, and during forest and grass fires. Caused by their hydrophobic nature, PAH basically accumulate in sediments from where they are slowly released into the groundwater. Although generally limited by the low water solubility of PAH, microbial degradation is one of the major mechanisms leading to the complete clean-up of PAH-contaminated sites. Whereas organisms and biochemical pathways responsible for the aerobic breakdown of PAH are well known, anaerobic PAH biodegradation is less understood; only a few anaerobic PAH degrading cultures have been described. We studied the anaerobic PAH degradation in a microcosm approach to enrich anaerobic PAH degraders. Anoxic groundwater and sediment samples were used as inoculum. Groundwater samples were purchased from the erstwhile gas works facility and a former wood impregnation site. In contrast, sources of sediment samples were a former coal refining area and an old fuel depot. Samples were incubated in anoxic mineral salt medium with naphthalene as sole carbon source and sulfate as terminal electron acceptor. Grown cultures were characterized by feeding with 13C-labeled naphthalene, 16S rRNA gene sequencing using an Illumina® approach, and functional proteome analyses. Finally, six enrichment cultures able to degrade naphthalene under anoxic conditions were established. First results point to a dominance of identified sequences affiliated to the freshwater sulfate-reducing strain N47, which is a known anaerobic naphthalene degrader, in four out of the six enrichments. In those enrichments, peptides related to the pathway of anoxic naphthalene degradation in N47 were abundant. Overall the data underlines the importance of Desulfobacteria for natural

  20. Epoxy Coenzyme A Thioester pathways for degradation of aromatic compounds.

    Science.gov (United States)

    Ismail, Wael; Gescher, Johannes

    2012-08-01

    Aromatic compounds (biogenic and anthropogenic) are abundant in the biosphere. Some of them are well-known environmental pollutants. Although the aromatic nucleus is relatively recalcitrant, microorganisms have developed various catabolic routes that enable complete biodegradation of aromatic compounds. The adopted degradation pathways depend on the availability of oxygen. Under oxic conditions, microorganisms utilize oxygen as a cosubstrate to activate and cleave the aromatic ring. In contrast, under anoxic conditions, the aromatic compounds are transformed to coenzyme A (CoA) thioesters followed by energy-consuming reduction of the ring. Eventually, the dearomatized ring is opened via a hydrolytic mechanism. Recently, novel catabolic pathways for the aerobic degradation of aromatic compounds were elucidated that differ significantly from the established catabolic routes. The new pathways were investigated in detail for the aerobic bacterial degradation of benzoate and phenylacetate. In both cases, the pathway is initiated by transforming the substrate to a CoA thioester and all the intermediates are bound by CoA. The subsequent reactions involve epoxidation of the aromatic ring followed by hydrolytic ring cleavage. Here we discuss the novel pathways, with a particular focus on their unique features and occurrence as well as ecological significance.

  1. Degradation of aromatic compounds and degradative pathway of 4-nitrocatechol by Ochrobactrum sp. B2.

    Science.gov (United States)

    Zhong, Qiuzan; Zhang, Haiyan; Bai, Wenqin; Li, Mei; Li, Baotong; Qiu, Xinghui

    2007-12-01

    The potential capacity of a soil methyl parathion-degrading bacterium strain, Ochrobactrum sp. B2, for degrading various aromatic compounds were investigated. The results showed B2 was capable of degrading diverse aromatic compounds, but amino-substituted benzene compounds, at a concentration up to 100 mg L(-1) in 4 days. B2 could use 4-nitrocatechol (4-NC) as a sole carbon and energy source with release of nitrite ion. The pathway for 4-NC degradation via 1,2,4-benzenetriol (BT) and hydroquinone (HQ) formation in B2 was proposed based on the identification and quantification of intermediates by gas chromatography-mass spectrometry (GC-MS), and high performance liquid chromatography (HPLC). Degradation studies carried out on a plasmid-cured derivative showed that the genes for 4-NC degradative pathway was plasmid-borne in B2, suggesting that B2 degrades both p-nitrophenol and 4-NC by enzymes encoded by genes on the same plasmid.

  2. Kynurenine pathway in psychosis: evidence of increased tryptophan degradation.

    LENUS (Irish Health Repository)

    Barry, Sandra

    2009-05-01

    The kynurenine pathway of tryptophan degradation may serve to integrate disparate abnormalities heretofore identified in research aiming to elucidate the complex aetiopathogenesis of psychotic disorders. Post-mortem brain tissue studies have reported elevated kynurenine and kynurenic acid in the frontal cortex and upregulation of the first step of the pathway in the anterior cingulate cortex of individuals with schizophrenia. In this study, we examined kynurenine pathway activity by measuring tryptophan breakdown, a number of pathway metabolites and interferon gamma (IFN-gamma), which is the preferential activator of the first-step enzyme, indoleamine dioxygenase (IDO), in the plasma of patients with major psychotic disorder. Plasma tryptophan, kynurenine pathway metabolites were measured using high-performance liquid chromatography (HPLC) in 34 patients with a diagnosis on the psychotic spectrum (schizophrenia or schizoaffective disorder) and in 36 healthy control subjects. IFN-gamma was measured using enzyme-linked immunosorbent assay (ELISA). The mean tryptophan breakdown index (kynurenine\\/tryptophan) was significantly higher in the patient group compared with controls (P < 0.05). IFN-gamma measures did not differ between groups (P = 0.23). No relationship was found between measures of psychopathology, symptom severity and activity in the first step in the pathway. A modest correlation was established between the tryptophan breakdown index and illness duration. These results provide evidence for kynurenine pathway upregulation, specifically involving the first enzymatic step, in patients with major psychotic disorder. Increased tryptophan degradation in psychoses may have potential consequences for the treatment of these disorders by informing the development of novel therapeutic compounds.

  3. Degradation of toluene-2,4-diamine by persulphate: kinetics, intermediates and degradation pathway.

    Science.gov (United States)

    Jiang, Yong-hai; Zhang, Jin-bao; Xi, Bei-dou; An, Da; Yang, Yu; Li, Ming-xiao

    2015-01-01

    In this study, the degradation of toluene-2,4-diamine (TDA) by persulphate (PS) in an aqueous solution at near-neutral pH was examined. The result showed that the degradation rate of TDA increased with increasing PS concentrations. The optimal dosage of PS in the reaction system was determined by efficiency indicator (I) coupling in the consumption of PS and decay half-life of TDA. Calculation showed that 0.74 mM of PS was the most effective dosage for TDA degradation, at that level the maximum I of 24.51 was obtained. PS can oxidize TDA for an extended reaction time period. Under neutral condition without activation, four degradation intermediates, 2,4-diamino-3-hydroxy-5-sulfonicacidtoluene, 2,4-diaminobenzaldehyde, 2,4-bis(vinylamino)benzaldehyde and 3,5-diamino-4-hydroxy-2-pentene, were identified by high-performance liquid chromatography-mass spectrometry. The tentative degradation pathway of TDA was proposed as well. It was found that hydroxyl radical played an important role in degradation of TDA with the activation of Fe2+, whereas PS anion and sulphate radicals were responsible for the degradation without activation of Fe2+.

  4. Degradation of phenazone in aqueous solution with ozone: influencing factors and degradation pathways.

    Science.gov (United States)

    Miao, Heng-Feng; Cao, Meng; Xu, Dan-Yao; Ren, Hong-Yan; Zhao, Ming-Xing; Huang, Zhen-Xing; Ruan, Wen-Quan

    2015-01-01

    Oxidation kinetics and degradation pathways of phenazone (an analgesic and antipyretic drug) upon reaction with O3 were investigated. Kinetic studies on degradation of phenazone were carried out under different operating conditions such as temperature, pH, anions and H2O2 addition. Results showed that the degradation followed the pseudo-first-order kinetic model. The reaction rate constant (kobs) of phenazone reached the maximum at 20 °C (9.653×10(-3) s(-1)). The presence of NO3(-) could enhance the degradation rate, while the addition of HCO3(-), SO4(2)(-), Cl(-) and the rise of pH showed negative effects on the ozonation of phenazone. H2O2 addition increased the phenazone degradation efficiency by 45.9% with the optimal concentration of 0.135 mM. Reaction by-products were evaluated by UPLC-Q-TOF-MS, which allowed the identification of a total of 10 by-products. The transformation pathways of phenazone ozonation consisted mainly of electrophilic addition and substitution, pyrazole ring opening, hydroxylation, dephenylization and coupling. The toxicity of these intermediate products showed that they are expected not to be more toxic than phenazone, with the exception of P7 (aniline) and P10 (1,5-dimethyl-4-((1-methyl-2-phenylhydrazinyl)methoxy)-2-phenyl-1H-pyrazol-3(2H)-one).

  5. Degradation Characterization of Thermal Interface Greases: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    DeVoto, Douglas J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Major, Joshua [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Paret, Paul P [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-08-03

    Thermal interface materials (TIMs) are used in power electronics packaging to minimize thermal resistance between the heat generating component and the heat sink. Thermal greases are one such class. The conformability and thin bond line thickness (BLT) of these TIMs can potentially provide low thermal resistance throughout the operation lifetime of a component. However, their performance degrades over time due to pump-out and dry-out during thermal and power cycling. The reliability performance of greases through operational cycling needs to be quantified to develop new materials with superior properties. NREL, in collaboration with DuPont, has performed thermal and reliability characterization of several commercially available thermal greases. Initial bulk and contact thermal resistance of grease samples were measured, and then the thermal degradation that occurred due to pump-out and dry-out during temperature cycling was monitored. The thermal resistances of five different grease materials were evaluated using NREL's steady-state thermal resistance tester based on the ASTM test method D5470. Greases were then applied, utilizing a 2.5 cm x 2.5 cm stencil, between invar and aluminum plates to compare the thermomechanical performance of the materials in a representative test fixture. Scanning Acoustic microscopy, thermal, and compositional analyses were performed periodically during thermal cycling from -40 degrees Celcius to 125 degrees Celcius. Completion of this characterization has allowed for a comprehensive evaluation of thermal greases both for their initial bulk and contact thermal performance, as well as their degradation mechanisms under accelerated thermal cycling conditions.

  6. Reversed-phase liquid chromatography with electrospray mass detection and 1H and 13C NMR characterization of new process-related impurities, including forced degradants of efavirenz: related substances correlated to the synthetic pathway.

    Science.gov (United States)

    Gadapayale, Kamalesh; Kakde, Rajendra; Sarma, V U M

    2015-01-01

    In this study, a stability-indicating reversed-phase liquid chromatographic electrospray mass spectrometric method was developed and validated for the determination of process-related impurities and forced degradants of Efavirenz in bulk drugs. Efavirenz was subjected to acid, alkaline hydrolysis, H2O2 oxidation, photolysis, and thermal stress. Significant degradation was observed during alkaline hydrolysis, and the degradants were isolated on a mass-based purification system and characterized by high-resolution mass spectrometry, positive electrospray ionization tandem mass spectrometry, and (1)H and (13)C NMR spectroscopy. Accurate mass measurement and NMR spectroscopy revealed the possible structure of process-related impurities and degradant under stress conditions. The acceptable separation was accomplished on Waters bondapak C18 column (250 mm × 4.6 mm; 5 μm), using 5 mM ammonium acetate and acetonitrile as a mobile phase in a gradient elution mode at a flow rate of 1.0 mL/min. The eluents were monitored by diode array detector at 247 nm and quantitation limits were obtained in the range of 0.1-2.5 μg/mL for Efavirenz, degradants, and process-related impurities. The liquid chromatography method was validated with respect to accuracy, precision, linearity, robustness, and limits of detection and quantification as per International Conference on Harmonization guidelines. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Characterization of thermally degraded energetic materials

    Energy Technology Data Exchange (ETDEWEB)

    Renlund, A.M.; Miller, J.C.; Trott, W.M.; Erickson, K.L.; Hobbs, M.L.; Schmitt, R.G.; Wellman, G.W.; Baer, M.R.

    1997-12-31

    Characterization of the damage state of a thermally degraded energetic material (EM) is a critical first step in understanding and predicting cookoff behavior. Unfortunately, the chemical and mechanical responses of heated EMs are closely coupled, especially if the EM is confined. The authors have examined several EMs in small-scale experiments (typically 200 mg) heated in both constant-volume and constant-load configurations. Fixtures were designed to minimize free volume and to contain gas pressures to several thousand psi. The authors measured mechanical forces or displacements that correlated to thermal expansion, phase transitions, material creep and gas pressurization as functions of temperature and soak time. In addition to these real-time measurements, samples were recovered for postmortem examination, usually with scanning electron microscopy (SEM) and chemical analysis. The authors present results on EMs (HMX and TATB), with binders (e.g., PBX 9501, PBX 9502, LX-14) and propellants (Al/AP/HTPB).

  8. Aerobic Degradation of Dinitrotoluenes and Pathway for Bacterial Degradation of 2,6-Dinitrotoluene

    OpenAIRE

    Nishino, Shirley F.; Paoli, George C.; Spain, Jim C.

    2000-01-01

    An oxidative pathway for the mineralization of 2,4-dinitrotoluene (2,4-DNT) by Burkholderia sp. strain DNT has been reported previously. We report here the isolation of additional strains with the ability to mineralize 2,4-DNT by the same pathway and the isolation and characterization of bacterial strains that mineralize 2,6-dinitrotoluene (2,6-DNT) by a different pathway. Burkholderia cepacia strain JS850 and Hydrogenophaga palleronii strain JS863 grew on 2,6-DNT as the sole source of carbon...

  9. Characterization of amygdalin-degrading Lactobacillus species.

    Science.gov (United States)

    Menon, R; Munjal, N; Sturino, J M

    2015-02-01

    Cyanogenic glycosides are phytotoxic secondary metabolites produced by some crop plants. The aim of this study was to identify lactic acid bacteria (LAB) capable of catabolizing amygdalin, a model cyanogenic glycoside, for use in the biodetoxification of amygdalin-containing foods and feeds. Amygdalin-catabolizing lactobacilli were characterized using a combination of cultivation-dependent and molecular assays. Lactobacillus paraplantarum and Lactobacillus plantarum grew robustly on amygdalin (Amg(+)), while other LAB species typically failed to catabolize amygdalin (Amg(-)). Interestingly, high concentrations of amygdalin and two of its metabolic derivatives (mandelonitrile and benzaldehyde) inhibited the growth of Lact. plantarum RENO 0093. The differential regulation of genes tentatively involved in cyanohydrin metabolism illustrated that the metabolism of amygdalin- and glucose-grown cultures also differed significantly. Amygdalin fermentation was a relatively uncommon phenotype among the LAB and generally limited to strains from the Lact. plantarum group. Phenotype microarrays (PM) enabled strain-level discrimination between closely related strains within a species and suggested that phenotypic differences might affect niche specialization. Amygdalin-degrading lactobacilli with practical application in the biodetoxification of amygdalin were characterized. These strains show potential for use as starter cultures to improve the safety of foods and feeds. © 2014 The Society for Applied Microbiology.

  10. Photocatalytic degradation of methylene blue with a nanocomposite system: synthesis, photocatalysis and degradation pathways.

    Science.gov (United States)

    Xia, Shengjie; Zhang, Lianyang; Pan, Guoxiang; Qian, Pingping; Ni, Zheming

    2015-02-21

    Three different composites, including a calcined FeOOH supported ZnAl layered double hydroxide (FeOOH-LDO), a calcined ZnAl layered double hydroxide (ZnAl-LDO) and a calcined ZnFeAl layered double hydroxide (ZnFeAl-LDO), were synthesized via a sol-gel method, and their activity for the visible light photocatalytic degradation of methylene blue (MB) was studied. The composites were characterized by PXRD, SEM, and BET techniques, confirming the formation of highly crystalline structures. The activity performance of MB degradation was in the following order: FeOOH-LDO (∼95%) > ZnFeAl-LDO (∼60%) > ZnAl-LDO (∼23%). In addition, a possible photocatalytic degradation reaction mechanism for MB was also proposed. Moreover, the frontier electron densities on the atoms of MB were calculated, which were in satisfactory agreement with the postulated mechanism.

  11. Hydrolytic and oxidative degradation of electrospun supramolecular biomaterials: In vitro degradation pathways.

    Science.gov (United States)

    Brugmans, M C P; Sӧntjens, S H M; Cox, M A J; Nandakumar, A; Bosman, A W; Mes, T; Janssen, H M; Bouten, C V C; Baaijens, F P T; Driessen-Mol, A

    2015-11-01

    The emerging field of in situ tissue engineering (TE) of load bearing tissues places high demands on the implanted scaffolds, as these scaffolds should provide mechanical stability immediately upon implantation. The new class of synthetic supramolecular biomaterial polymers, which contain non-covalent interactions between the polymer chains, thereby forming complex 3D structures by self assembly. Here, we have aimed to map the degradation characteristics of promising (supramolecular) materials, by using a combination of in vitro tests. The selected biomaterials were all polycaprolactones (PCLs), either conventional and unmodified PCL, or PCL with supramolecular hydrogen bonding moieties (either 2-ureido-[1H]-pyrimidin-4-one or bis-urea units) incorporated into the backbone. As these materials are elastomeric, they are suitable candidates for cardiovascular TE applications. Electrospun scaffold strips of these materials were incubated with solutions containing enzymes that catalyze hydrolysis, or solutions containing oxidative species. At several time points, chemical, morphological, and mechanical properties were investigated. It was demonstrated that conventional and supramolecular PCL-based polymers respond differently to enzyme-accelerated hydrolytic or oxidative degradation, depending on the morphological and chemical composition of the material. Conventional PCL is more prone to hydrolytic enzymatic degradation as compared to the investigated supramolecular materials, while, in contrast, the latter materials are more susceptible to oxidative degradation. Given the observed degradation pathways of the examined materials, we are able to tailor degradation characteristics by combining selected PCL backbones with additional supramolecular moieties. The presented combination of in vitro test methods can be employed to screen, limit, and select biomaterials for pre-clinical in vivo studies targeted to different clinical applications.

  12. Activity, biomass and composition of microbial communities and their degradation pathways in exposed propazine soil.

    Science.gov (United States)

    Jiang, Chen; Lu, Yi Chen; Xu, Jiang Yan; Song, Yang; Song, Yue; Zhang, Shu Hao; Ma, Li Ya; Lu, Feng Fan; Wang, Ya Kun; Yang, Hong

    2017-11-01

    Propazine is a s-triazine herbicide widely used for controlling weeds for crop production. Its persistence and contamination in environment nagatively affect crop growth and food safety. Elimination of propazine residues in the environment is critical for safe crop production. This study identified a microbial community able to degrade propazine in a farmland soil. About 94% of the applied propazine was degraded within 11 days of incubation when soil was treated with 10mgkg(-1) propazine as the initial concentration. The process was accompanied by increased microbial biomass and activities of soil enzymes. Denaturing gradient gel electrophoresis (DGGE) revealed multiple bacterial strains in the community as well as dynamic change of the composition of microbial community with a reduced microbial diversity (H' from 3.325 to 2.78). Tracking the transcript level of degradative genes AtzB, AtzC and TrzN showed that these genes were induced by propazine and played important roles in the degradation process. The activities of catalase, dehydrogenase and phenol oxidase were stimulated by propazine exposure. Five degradation products (hydroxyl-, methylated-, dimeric-propazine, ammeline and ammelide) were characterized by UPLC-MS(2), revealing a biodegradation of propazine in soil. Several novel methylated and dimeric products of propazine were characterized in thepropazine-exposed soil. These data help understand the pathway, detailed mechanism and efficiency of propazine biodegradation in soil under realistic field condition. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. ARTS and Siah Collaborate in a Pathway for XIAP Degradation

    Science.gov (United States)

    Garrison, Jason B.; Correa, Ricardo G.; Gerlic, Motti; Yip, Kenneth W.; Krieg, Andreas; Tamble, Craig M.; Shi, Ranxin; Welsh, Kate; Duggineni, Srinivas; Huang, Ziwei; Ren, Keqin; Du, Chunying; Reed, John C.

    2010-01-01

    SUMMARY ARTS (Apoptosis-Related protein in the TGF-β Signaling pathway) is a mitochondrial protein that binds XIAP (X-linked Inhibitor of Apoptosis Protein) upon entering the cytosol, thus promoting cell death. Expression of ARTS is lost in some malignancies. Here we show that ARTS binds to XIAP at BIR1, a domain distinct from the caspase-binding sites. Furthermore, ARTS interacts with the E3 ligase Siah-1 (seven in absentia homolog 1) to induce ubiquitination and degradation of XIAP. Cells lacking either Siah or ARTS contain higher steady-state levels of XIAP. Thus, ARTS serves as an adapter to bridge Siah-1 to XIAP, targeting it for destruction. PMID:21185211

  14. Thermally induced degradation pathways of three different antibody-based drug development candidates.

    Science.gov (United States)

    Fincke, Anja; Winter, Jonas; Bunte, Thomas; Olbrich, Carsten

    2014-10-01

    Protein-based medicinal products are prone to undergo a variety of chemical and physical degradation pathways. One of the most important exogenous stress condition to consider during manufacturing, transport and storage processes is temperature, because antibody-based therapeutics are only stable in a limited temperature range. In this study, three different formats of antibody-based molecules (IgG1, a bispecific scFv and a fab fragment) were exposed to thermal stress conditions occurring during transport and storage. For evaluation, an analytical platform was developed for the detection and characterization of relevant degradation pathways of different antibody-based therapeutics. The effect of thermal stress conditions on the stability of the three antibody-based formats was therefore investigated using visual inspection, different spectroscopic measurements, dynamic light scattering (DLS), differential scanning calorimetry (DSC), electrophoresis, asymmetric flow field-flow fractionation (AF4) and surface plasmon resonance technology (SPR). In summary, thermal stress led to heterogeneous chemical and physical degradation pathways of all three antibody-based formats used. In addition, identical exogenous stress conditions resulted in different kinds and levels of aggregates and fragmentation products. This knowledge is fundamental for a systematic and successful stabilization of protein-based therapeutics by the use of formulation additives.

  15. Biotransformation of nitrobenzene by bacteria containing toluene degradative pathways

    Energy Technology Data Exchange (ETDEWEB)

    Haigler, B.E.; Spain, J.C. (Air Force Civil Engineering Support Agency, Tyndall AFB, FL (United States))

    1991-11-01

    Nonpolar nitroaromatic compounds have been considered resistant to attack by oxygenases because of the electron withdrawing properties of the nitro group. The authors have investigate the ability of seven bacterial strains containing toluene degradative pathways to oxidize nitrobenzene. Cultures were induced with toluene vapor prior to incubation with nitrobenzene, and products were identified by high-performance liquid chromatography and gas chromatography-mass spectrometry. Pseudomonas cepacia G4 and a strain of Pseudomonas harboring the TOL plasmid (pTN2) did not transform nitrobenzene. Cells of Pseudomonas putida F1 and Pseudomonas sp. strain JS150 converted nitrobenzene to 3-nitrocatechol. Transformation of nitrobenzene in the presence of {sup 18}O{sub 2} indicated that the reaction in JS150 involved the incorporation of both atoms of oxygen in the 3-nitrocatechol, which suggests a dioxygenase mechanism. P. putida 39/D, a mutant strain of P. putida F1, converted nitrobenzene to a compound tentatively identified as cis-1, 2-dihydroxy-3-nitrocyclohexa-3, 5-diene. This compound was rapidly converted to 3-nitrocatechol by cells of strain JS150. Cultures of Pseudomonas mendocina KR-1 converted nitrobenzene to a mixture of 3- and 4-nitrophenol (10 and 63%, respectively). Pseudomonas pickettii PKO1 converted nitrobenzene to 3- and 4-nitrocatechol via 3- and 4-nitrophenol. The nitrocatechols were slowly degraded to unidentified metabolites. Nitrobenzene did not serve as an inducer for the enzymes that catalyzed its oxidation.

  16. Fenton degradation of Cartap hydrochloride: identification of the main intermediates and the degradation pathway.

    Science.gov (United States)

    Tian, Kaixun; Ming, Cuixiang; Dai, Youzhi; Honore Ake, Kouassi Marius

    2015-01-01

    The advanced oxidation of Cartap hydrochloride (Cartap) promoted by the Fenton system in an aqueous medium was investigated. Based on total organic carbon, chemical oxygen demand and high-performance liquid chromatography, the oxidation of Cartap is quite efficient by the Fenton system. Its long chain is easily destroyed, but the reaction does not proceed to complete mineralization. Ion chromatography detection indicated the formation of acetic acid, propionic acid, formic acid, nitrous acid and sulfuric acid in the reaction mixtures. Further evidence of nitrogen monoxide and sulfur dioxide formation was obtained by using a flue gas analyzer. Monitoring by gas chromatograph-mass spectrometer demonstrated the formation of oxalic acid, ethanol, carbon dioxide, and L-alanine ethylamide. Based on these experimental results, plausible degradation pathways for Cartap mineralization in an aqueous medium by the Fenton system are proposed.

  17. A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells.

    Science.gov (United States)

    Huber, S C; Akazawa, T

    1986-08-01

    Enzymes of sucrose degradation and glycolysis in cultured sycamore (Acer pseudoplatanus L.) cells were assayed and characterized in crude extracts and after partial purification, in an attempt to identify pathways for sucrose catabolism. Desalted cell extracts contained similar activities (20-40 nanomoles per milligram protein per minute) of sucrose synthase, neutral invertase, glucokinase, fructokinase, phosphofructokinase, and UDPglucose pyrophosphorylase (assayed with 2 micromolar pyrophosphate (PPi). PPi-linked phosphofructokinase activity was virtually dependent upon fructose 2,6-bisphosphate, and the maximum activity exceeded that of ATP-linked phosphofructokinase. Hexokinase activity, with glucose as substrate, was highly specific for ATP, whereas fructokinase activity was relatively nonspecific. At 1 millimolar nucleoside triphosphate, fructokinase activity decreased in the order: UTP > ATP > CTP > GTP. We propose two pathways for sucrose degradation. One involves invertase action, followed by classical glycolysis of hexose sugars, and the other is a novel pathway initiated by sucrose synthase. The K(m) for sucrose of sucrose synthase was severalfold lower than that of neutral invertase (15 versus 65 millimolar), which may determine carbon partitioning between the two pathways. The sucrose synthase pathway proposed involves cycling of uridylates and PPi. UDPglucose pyrophosphorylase, which is shown to be an effective ;PPi-scavenger,' would consume PPi and form UTP. The UTP could be then utilized in the UTP-linked fructokinase reaction, thereby forming UDP for sucrose synthase. The source of PPi is postulated to arise from the back reaction of PPi-linked phosphofructokinase. Sycamore cells contained a substantial endogenous pool of PPi (about 3 nanomoles per gram fresh weight, roughly 1/10 the amount of ATP in these cells), and sufficient fructose 2,6-bisphosphate (0.09 nanomole per gram fresh weight) to activate the PPi-linked phosphofructokinase. Possible

  18. Mechanochemical degradation of tetrabromobisphenol A: Performance, products and pathway

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Kunlun; Huang, Jun; Zhang, Wang; Yu, Yunfei; Deng, Shubo [State Key Joint Laboratory of Environment Simulation and Pollution Control (SKLESPC), School of Environment, POPs Research Center, Tsinghua University, Beijing 100084 (China); Yu, Gang, E-mail: yg-den@tsinghua.edu.cn [State Key Joint Laboratory of Environment Simulation and Pollution Control (SKLESPC), School of Environment, POPs Research Center, Tsinghua University, Beijing 100084 (China)

    2012-12-15

    Highlights: Black-Right-Pointing-Pointer Fe + SiO{sub 2} shows better performance than CaO in mechanochemical destruction of TBBPA. Black-Right-Pointing-Pointer Nonhazardous inorganic carbon and soluble bromide were the final products. Black-Right-Pointing-Pointer Raman and FTIR imply the generation of inorganic carbon and removal of bromine atom. Black-Right-Pointing-Pointer Tri-BBPA, bi-BBPA, mono-BBPA, BPA were the main intermediates during ball milling. Black-Right-Pointing-Pointer The bromine was balanced and the degradation pathway was proposed. - Abstract: Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant (BFR), which has received more and more concerns due to its high lipophilicity, persistency and endocrine disrupting property in the environment. Considering the possible need for the safe disposal of TBBPA containing wastes in the future, the potential of mechanochemical (MC) destruction as a promising non-combustion technology was investigated in this study. TBBPA was co-ground with calcium oxide (CaO) or the mixture of iron powder and quartz sand (Fe + SiO{sub 2}) in a planetary ball mill at room temperature. The method of Fe + SiO{sub 2} destructed over 98% of initial TBBPA after 3 h and acquired 95% debromination rate after 5 h, which showed a better performance than the CaO method. Raman spectra and Fourier transform infrared spectroscopy (FTIR) demonstrated the generation of inorganic carbon with the disappearance of benzene ring and C-Br bond, indicating the carbonization and debromination process during mechanochemical reaction. LC-MS-MS screening showed that the intermediates of the treatment with Fe + SiO{sub 2} were tri-, bi-, mono-brominated BPA, BPA and other fragments. Finally all the intermediates were also destroyed after 5 h grinding. The bromine balance was calculated and a possible reaction pathway was proposed.

  19. Unraveling the Specific Regulation of the Central Pathway for Anaerobic Degradation of 3-Methylbenzoate*

    Science.gov (United States)

    Juárez, Javier F.; Liu, Huixiang; Zamarro, María T.; McMahon, Stephen; Liu, Huanting; Naismith, James H.; Eberlein, Christian; Boll, Matthias; Carmona, Manuel; Díaz, Eduardo

    2015-01-01

    The mbd cluster encodes the anaerobic degradation of 3-methylbenzoate in the β-proteobacterium Azoarcus sp. CIB. The specific transcriptional regulation circuit that controls the expression of the mbd genes was investigated. The PO, PB1, and P3R promoters responsible for the expression of the mbd genes, their cognate MbdR transcriptional repressor, as well as the MbdR operator regions (ATACN10GTAT) have been characterized. The three-dimensional structure of MbdR has been solved revealing a conformation similar to that of other TetR family transcriptional regulators. The first intermediate of the catabolic pathway, i.e. 3-methylbenzoyl-CoA, was shown to act as the inducer molecule. An additional MbdR-dependent promoter, PA, which contributes to the expression of the CoA ligase that activates 3-methylbenzoate to 3-methylbenzoyl-CoA, was shown to be necessary for an efficient induction of the mbd genes. Our results suggest that the mbd cluster recruited a regulatory system based on the MbdR regulator and its target promoters to evolve a distinct central catabolic pathway that is only expressed for the anaerobic degradation of aromatic compounds that generate 3-methylbenzoyl-CoA as the central metabolite. All these results highlight the importance of the regulatory systems in the evolution and adaptation of bacteria to the anaerobic degradation of aromatic compounds. PMID:25795774

  20. Chemical modification and degradation of atrazine in Medicago sativa through multiple pathways.

    Science.gov (United States)

    Zhang, Jing Jing; Lu, Yi Chen; Yang, Hong

    2014-10-08

    Atrazine is a member of the triazine herbicide family intensively used to control weeds for crop production. In this study, atrazine residues and its degraded products in alfalfa (Medicago sativa) were characterized using UPLC-TOF-MS/MS. Most of atrazine absorbed in plants was found as chemically modified derivatives like deisopropylated atrazine (DIA), dehydrogenated atrazine (DHA), or methylated atrazine (MEA), and some atrazine derivatives were conjugated through different functional groups such as sugar, glutathione, and amino acids. Interestingly, the specific conjugates DHA+hGSH (homoglutathione) and MEA-HCl+hGSH in alfalfa were detected. These results suggest that atrazine in alfalfa can be degraded through different pathways. The increased activities of glycosyltransferase and glutathione S-transferase were determined to support the atrazine degradation models. The outcome of the work uncovered the detailed mechanism for the residual atrazine accumulation and degradation in alfalfa and will help to evaluate whether the crop is suitable to be cultivated in the atrazine-polluted soil.

  1. Unusual starch degradation pathway via cyclodextrins in the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus strain 7324.

    Science.gov (United States)

    Labes, Antje; Schönheit, Peter

    2007-12-01

    The hyperthermophilic archaeon Archaeoglobus fulgidus strain 7324 has been shown to grow on starch and sulfate and thus represents the first sulfate reducer able to degrade polymeric sugars. The enzymes involved in starch degradation to glucose 6-phosphate were studied. In extracts of starch-grown cells the activities of the classical starch degradation enzymes, alpha-amylase and amylopullulanase, could not be detected. Instead, evidence is presented here that A. fulgidus utilizes an unusual pathway of starch degradation involving cyclodextrins as intermediates. The pathway comprises the combined action of an extracellular cyclodextrin glucanotransferase (CGTase) converting starch to cyclodextrins and the intracellular conversion of cyclodextrins to glucose 6-phosphate via cyclodextrinase (CDase), maltodextrin phosphorylase (Mal-P), and phosphoglucomutase (PGM). These enzymes, which are all induced after growth on starch, were characterized. CGTase catalyzed the conversion of starch to mainly beta-cyclodextrin. The gene encoding CGTase was cloned and sequenced and showed highest similarity to a glucanotransferase from Thermococcus litoralis. After transport of the cyclodextrins into the cell by a transport system to be defined, these molecules are linearized via a CDase, catalyzing exclusively the ring opening of the cyclodextrins to the respective maltooligodextrins. These are degraded by a Mal-P to glucose 1-phosphate. Finally, PGM catalyzes the conversion of glucose 1-phosphate to glucose 6-phosphate, which is further degraded to pyruvate via the modified Embden-Meyerhof pathway.

  2. Characterization of the Oxidative Degradation Product of Darunavir by LC-MS/MS

    Science.gov (United States)

    Yamjala, Karthik; Atukuri, Jeevitha; Nagappan, Krishnaveni; Halekote Shivaraju, Nivedeetha; Subramania Nainar, Meyyanathan

    2015-01-01

    A rapid, selective, and reliable LC-MSn method has been developed and validated for the isolation and structural characterization of the degradation product of darunavir (DRV). DRV, an HIV-1 protease inhibitor, was subjected to intrinsic oxidative stress conditions using 30% hydrogen peroxide and the degradation profile was studied. The oxidative degradation of DRV resulted in one degradation product. The unknown degradation product was separated on a Hibar Purospher C18 (250 mm × 4.6 mm; 5 µm) column by using 0.01 M ammonium formate (pH 3.0) and acetonitrile as mobile phase in the ratio of 50:50, v/v. The eluents were monitored at 263 nm using a UV detector. The isolated degradation product was characterized by UPLC-Q-TOF and its fragmentation pathway was proposed. The proposed structure of the degradation product was confirmed by HRMS analysis. The developed stability-indicating LC method was validated with respect to accuracy, precision, specificity/selectivity, and linearity. No prior reports were found in the literature about the oxidative degradation behavior of DRV. PMID:26839843

  3. Cycle Inhibiting Factors (Cifs): Cyclomodulins That Usurp the Ubiquitin-Dependent Degradation Pathway of Host Cells

    Science.gov (United States)

    Taieb, Frédéric; Nougayrède, Jean-Philippe; Oswald, Eric

    2011-01-01

    Cycle inhibiting factors (Cifs) are type III secreted effectors produced by diverse pathogenic bacteria. Cifs are “cyclomodulins” that inhibit the eukaryotic host cell cycle and also hijack other key cellular processes such as those controlling the actin network and apoptosis. This review summarizes current knowledge on Cif since its first characterization in enteropathogenic Escherichia coli, the identification of several xenologues in distant pathogenic bacteria, to its structure elucidation and the recent deciphering of its mode of action. Cif impairs the host ubiquitin proteasome system through deamidation of ubiquitin or the ubiquitin-like protein NEDD8 that regulates Cullin-Ring-ubiquitin Ligase (CRL) complexes. The hijacking of the ubiquitin-dependent degradation pathway of host cells results in the modulation of various cellular functions such as epithelium renewal, apoptosis and immune response. Cif is therefore a powerful weapon in the continuous arm race that characterizes host-bacteria interactions. PMID:22069713

  4. Cycle inhibiting factors (cifs): cyclomodulins that usurp the ubiquitin-dependent degradation pathway of host cells.

    Science.gov (United States)

    Taieb, Frédéric; Nougayrède, Jean-Philippe; Oswald, Eric

    2011-04-01

    Cycle inhibiting factors (Cifs) are type III secreted effectors produced by diverse pathogenic bacteria. Cifs are "cyclomodulins" that inhibit the eukaryotic host cell cycle and also hijack other key cellular processes such as those controlling the actin network and apoptosis. This review summarizes current knowledge on Cif since its first characterization in enteropathogenic Escherichia coli, the identification of several xenologues in distant pathogenic bacteria, to its structure elucidation and the recent deciphering of its mode of action. Cif impairs the host ubiquitin proteasome system through deamidation of ubiquitin or the ubiquitin-like protein NEDD8 that regulates Cullin-Ring-ubiquitin Ligase (CRL) complexes. The hijacking of the ubiquitin-dependent degradation pathway of host cells results in the modulation of various cellular functions such as epithelium renewal, apoptosis and immune response. Cif is therefore a powerful weapon in the continuous arm race that characterizes host-bacteria interactions.

  5. Cycle Inhibiting Factors (Cifs: Cyclomodulins That Usurp the Ubiquitin-Dependent Degradation Pathway of Host Cells

    Directory of Open Access Journals (Sweden)

    Eric Oswald

    2011-03-01

    Full Text Available Cycle inhibiting factors (Cifs are type III secreted effectors produced by diverse pathogenic bacteria. Cifs are “cyclomodulins” that inhibit the eukaryotic host cell cycle and also hijack other key cellular processes such as those controlling the actin network and apoptosis. This review summarizes current knowledge on Cif since its first characterization in enteropathogenic Escherichia coli, the identification of several xenologues in distant pathogenic bacteria, to its structure elucidation and the recent deciphering of its mode of action. Cif impairs the host ubiquitin proteasome system through deamidation of ubiquitin or the ubiquitin-like protein NEDD8 that regulates Cullin-Ring-ubiquitin Ligase (CRL complexes. The hijacking of the ubiquitin-dependent degradation pathway of host cells results in the modulation of various cellular functions such as epithelium renewal, apoptosis and immune response. Cif is therefore a powerful weapon in the continuous arm race that characterizes host-bacteria interactions.

  6. Complementary roles of intracellular and pericellular collagen degradation pathways in vivo

    DEFF Research Database (Denmark)

    Wagenaar-Miller, Rebecca A; Engelholm, Lars H; Gavard, Julie

    2007-01-01

    these two pathways is unclear and even controversial. Here we show that intracellular and pericellular collagen turnover pathways have complementary roles in vivo. Individual deficits in intracellular collagen degradation (urokinase plasminogen activator receptor-associated protein/Endo180 ablation......Collagen degradation is essential for cell migration, proliferation, and differentiation. Two key turnover pathways have been described for collagen: intracellular cathepsin-mediated degradation and pericellular collagenase-mediated degradation. However, the functional relationship between......) or pericellular collagen degradation (membrane type 1-matrix metalloproteinase ablation) were compatible with development and survival. Their combined deficits, however, synergized to cause postnatal death by severely impairing bone formation. Interestingly, this was mechanistically linked to the proliferative...

  7. Degradation pathway of 2-chloroethanol in Pseudomonas stutzeri strain JJ under denitrifying conditions

    NARCIS (Netherlands)

    Dijk, J.A.; Gerritse, J.; Schraa, G.; Stams, A.J.M.

    2004-01-01

    The pathway of 2-chloroethanol degradation in the denitrifying Pseudomonas stutzeri strain JJ was investigated. In cell-free extracts, activities of a phenazine methosulfate (PMS)-dependent chloroethanol dehydrogenase, an NAD-dependent chloroacetaldehyde dehydrogenase, and a chloroacetate dehalogena

  8. [Characterization of a thermophilic Geobacillus strain DM-2 degrading hydrocarbons].

    Science.gov (United States)

    Liu, Qing-kun; Wang, Jun; Li, Guo-qiang; Ma, Ting; Liang, Feng-lai; Liu, Ru-lin

    2008-12-01

    A thermophilic Geobacillus strain DM-2 from a deep-subsurface oil reservoir was investigated on its capability of degrading crude oil under various conditions as well as its characters on degrading hydrocarbons in optimal conditions. The results showed that Geobacillus strain DM-2 was able to degrade crude oil under anoxic wide-range conditions with pH ranging from 4.0 to 10.0, high temperature in the range of 45-70 degrees C and saline concentration ranging from 0.2% to 3.0%. Furthermore, the optimal temperature and pH value for utilizing hydrocarbons by the strain were 60 degrees C and 7.0, respectively. Under such optimal conditions, the strain utilized liquid paraffine emulsified by itself as its carbon source for growth; further analysis by gas chromatography (GC) and infrared absorption spectroscopy demonstrated that it was able to degrade n-alkanes (C14-C30), branched-chain alkanes and aromatic hydrocarbons in crude oil and could also utilize long-chain n-alkanes from C16 to C36, among of which the degradation efficiency of C28 was the highest, up to 88.95%. One metabolite of the strain oxidizing alkanes is fatty acid.While utilizing C16 as carbon source for 5 d, only one fatty acid-acetic acid was detected by HPLC and MS as the product, with the amount of 0.312 g/L, which indicated that it degraded n-alkanes with pathway of inferior terminal oxidation,and then followed by a beta-oxidation pathway. Due to its characters of efficient emulsification, high-performance degradation of hydrocarbons and fatty-acid production under high temperature and anoxic condition, the strain DM-2 may be potentially applied to oil-waste treatment and microbial enhanced heavy oil recovery in extreme conditions.

  9. Biochemical and physiological characterisation of the purine degradation pathway in plants

    OpenAIRE

    Werner, Andrea

    2013-01-01

    Plant growth is often limited by nitrogen availability in the soil. Not only do plants depend on efficient nitrogen uptake, they also require effective means to internally redistribute nitrogen during every stage of development. The purine degradation pathway contributes to this nitrogen recycling in plants. In tropical legumes it is also of central importance to the plants’ nitrogen supply under nitrogen-fixing conditions. This is the first time that the complete ureide degradation pathway h...

  10. Degradation of diclofenac by UV-activated persulfate process: Kinetic studies, degradation pathways and toxicity assessments.

    Science.gov (United States)

    Lu, Xian; Shao, Yisheng; Gao, Naiyun; Chen, Juxiang; Zhang, Yansen; Xiang, Huiming; Guo, Youluo

    2017-03-21

    Diclofenac (DCF) is the frequently detected non-steroidal pharmaceuticals in the aquatic environment. In this study, the degradation of DCF was evaluated by UV-254nm activated persulfate (UV/PS). The degradation of DCF followed the pseudo first-order kinetics pattern. The degradation rate constant (kobs) was accelerated by UV/PS compared to UV alone and PS alone. Increasing the initial PS dosage or solution pH significantly enhanced the degradation efficiency. Presence of various natural water constituents had different effects on DCF degradation, with an enhancement or inhibition in the presence of inorganic anions (HCO3(-) or Cl(-)) and a significant inhibition in the presence of NOM. In addition, preliminary degradation mechanisms and major products were elucidated using LC-MS/MS. Hydroxylation, decarbonylation, ring-opening and cyclation reaction involving the attack of SO4(•)(-) or other substances, were the main degradation mechanism. TOC analyzer and Microtox bioassay were employed to evaluate the mineralization and cytotoxicity of solutions treated by UV/PS at different times, respectively. Limited elimination of TOC (32%) was observed during the mineralization of DCF. More toxic degradation products and their related intermediate species were formed, and the UV/PS process was suitable for removing the toxicity. Of note, longer degradation time may be considered for the final toxicity removal.

  11. Degradation Pathway for Eplerenone by Validated Stability Indicating UP-LC Method

    OpenAIRE

    Kondru Sudhakar Babu; Venkataramanna Madireddy; Venkata Somaraju Indukuri

    2012-01-01

    Degradation pathway for eplerenone is established as per ICH recommendations by validated and stability-indicating reverse phase liquid chromatographic method. Eplerenone is subjected to stress conditions of acid, base, oxidation, and thermal and photolysis. Significant degradation is observed in acid and base stress conditions. Four impurities are studied and the major degradant (RRT about 0.31) was identified by LC-MS and spectral analysis. The stress samples are assayed against a qualified...

  12. Characterization of degradation products of amorphous and polymorphic forms of clopidogrel bisulphate under solid state stress conditions

    DEFF Research Database (Denmark)

    Raijada, Dhara K; Prasad, Bhagwat; Paudel, Amrit

    2010-01-01

    The present study deals with the stress degradation studies on amorphous and polymorphic forms of clopidogrel bisulphate. The objective was to characterize the degradation products and postulate mechanism of decomposition of the drug under solid state stress conditions. For that, amorphous form...... degradation products were observed under various stress conditions. The structures of all of them were elucidated using LC-MS/TOF and LC-MS(n) studies. While one matched the known hydrolytic decomposition product of the drug in solution, seven others were new. The postulated degradation pathway and mechanism...

  13. Characterization of Methane Degradation and Methane-Degrading Microbes in Alaska Coastal Water

    Energy Technology Data Exchange (ETDEWEB)

    Kirchman, David L. [Univ. of Delaware, Lewes, DE (United States)

    2012-03-29

    The net flux of methane from methane hydrates and other sources to the atmosphere depends on methane degradation as well as methane production and release from geological sources. The goal of this project was to examine methane-degrading archaea and organic carbon oxidizing bacteria in methane-rich and methane-poor sediments of the Beaufort Sea, Alaska. The Beaufort Sea system was sampled as part of a multi-disciplinary expedition (Methane in the Arctic Shelf or MIDAS) in September 2009. Microbial communities were examined by quantitative PCR analyses of 16S rRNA genes and key methane degradation genes (pmoA and mcrA involved in aerobic and anaerobic methane degradation, respectively), tag pyrosequencing of 16S rRNA genes to determine the taxonomic make up of microbes in these sediments, and sequencing of all microbial genes (metagenomes ). The taxonomic and functional make-up of the microbial communities varied with methane concentrations, with some data suggesting higher abundances of potential methane-oxidizing archaea in methane-rich sediments. Sequence analysis of PCR amplicons revealed that most of the mcrA genes were from the ANME-2 group of methane oxidizers. According to metagenomic data, genes involved in methane degradation and other degradation pathways changed with sediment depth along with sulfate and methane concentrations. Most importantly, sulfate reduction genes decreased with depth while the anaerobic methane degradation gene (mcrA) increased along with methane concentrations. The number of potential methane degradation genes (mcrA) was low and inconsistent with other data indicating the large impact of methane on these sediments. The data can be reconciled if a small number of potential methane-oxidizing archaea mediates a large flux of carbon in these sediments. Our study is the first to report metagenomic data from sediments dominated by ANME-2 archaea and is one of the few to examine the entire microbial assemblage potentially involved in

  14. MAPKs are essential upstream signaling pathways in proteolytic cartilage degradation--divergence in pathways leading to aggrecanase and MMP-mediated articular cartilage degradation

    DEFF Research Database (Denmark)

    Sondergaard, B-C; Schultz, N; Madsen, S H;

    2010-01-01

    Matrix metalloproteinases (MMPs) and aggrecanases are essential players in cartilage degradation. However, the signaling pathways that results in MMP and/or aggrecanase synthesis and activation are not well understood. We investigated the molecular events leading to MMP- and aggrecanase-mediated ...

  15. Bioenergetics and pathway of acid blue 113 degradation by Staphylococcus lentus.

    Science.gov (United States)

    Sekar, Sudharshan; Mahadevan, Surianarayanan; Shanmugam, Bhuvanesh Kumar; Mandal, Asit Baran

    2012-01-01

    Bioreaction calorimetric studies of degradation of the dye acid blue 113 by Staphylococcus lentus are reported for the first time. The heat released during the dye degradation process can be successfully measured using reaction calorimeter. Power time and oxygen uptake rate (OUR) profile followed each other suggesting that heat profiles could monitor the progress of the dye degradation in biocalorimetry. The shifts observed in power-time profile indicated three distinct phases of the bioprocess indicating simultaneous utilization of glucose (primary) and dye (secondary carbon source). Secretion of azoreductase enzyme enhanced the degradation process. Optimization of aeration and agitation rates was observed to be vital to efficient dye degradation. The degradative pathway for acid blue 113 by S. lentus was delineated via high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR), and gas chromatography coupled with mass spectrometry (GC-MS) analyses. Interestingly the products of degradation were found to have low toxicity, as per cytotoxicity measurements.

  16. Degradation pathway of malachite green in a novel dual-tank photoelectrochemical catalytic reactor

    Energy Technology Data Exchange (ETDEWEB)

    Diao, Zenghui [Department of Environmental Engineering, Jinan University, Guangzhou 510630 (China); School of Environmental Science and Engineering, Sun Yat-Sen University, Guangzhou 510275 (China); Li, Mingyu, E-mail: tlimy@jnu.edu.cn [Department of Environmental Engineering, Jinan University, Guangzhou 510630 (China); Zeng, Fanyin; Song, Lin [Department of Environmental Engineering, Jinan University, Guangzhou 510630 (China); Qiu, Rongliang [School of Environmental Science and Engineering, Sun Yat-Sen University, Guangzhou 510275 (China)

    2013-09-15

    Highlights: • A novel dual-tank photoelectrochemical catalytic reactor was designed. • Malachite green degraded in bipolar double-effect mode. • Salt bridge replaced by a cation exchange membrane in the reactor. • Degradation pathways of malachite green in the cathode and anode tanks were similar. -- Abstract: A novel dual-tank photoelectrochemical catalytic reactor was designed to investigate the degradation pathway of malachite green. A thermally formed TiO{sub 2}/Ti thin film electrode was used as photoanode, graphite was used as cathode, and a saturated calomel electrode was employed as the reference electrode in the reactor. In the reactor, the anode and cathode tanks were connected by a cation exchange membrane. Results showed that the decolorization ratio of malachite green in the anode and cathode was 98.5 and 96.5% after 120 min, respectively. Malachite green in the two anode and cathode tanks was oxidized, achieving the bipolar double effect. Malachite green in both the anode and cathode tanks exhibited similar catalytic degradation pathways. The double bond of the malachite green molecule was attacked by strong oxidative hydroxyl radicals, after which the organic compound was degraded by the two pathways into 4,4-bis(dimethylamino) benzophenone, 4-(dimethylamino) benzophenone, 4-(dimethylamino) phenol, and other intermediate products. Eventually, malachite green was degraded into oxalic acid as a small molecular organic acid, which was degraded by processes such as demethylation, deamination, nitration, substitution, addition, and other reactions.

  17. Physiology of deletion mutants in the anaerobic β-myrcene degradation pathway in Castellaniella defragrans

    Directory of Open Access Journals (Sweden)

    Lüddeke Frauke

    2012-09-01

    Full Text Available Abstract Background Monoterpenes present a large and versatile group of unsaturated hydrocarbons of plant origin with widespread use in the fragrance as well as food industry. The anaerobic β-myrcene degradation pathway in Castellaniella defragrans strain 65Phen differs from well known aerobic, monooxygenase-containing pathways. The initial enzyme linalool dehydratase-isomerase ldi/LDI catalyzes the hydration of β-myrcene to (S-(+-linalool and its isomerization to geraniol. A high-affinity geraniol dehydrogenase geoA/GeDH and a geranial dehydrogenase geoB/GaDH contribute to the formation of geranic acid. A genetic system was for the first time applied for the betaproteobacterium to prove in vivo the relevance of the linalool dehydratase-isomerase and the geraniol dehydrogenase. In-frame deletion cassettes were introduced by conjugation and two homologous recombination events. Results Polar effects were absent in the in-frame deletion mutants C. defragrans Δldi and C. defragrans ΔgeoA. The physiological characterization of the strains demonstrated a requirement of the linalool dehydratase-isomerase for growth on acyclic monoterpenes, but not on cyclic monoterpenes. The deletion of geoA resulted in a phenotype with hampered growth rate on monoterpenes as sole carbon and energy source as well as reduced biomass yields. Enzyme assays revealed the presence of a second geraniol dehydrogenase. The deletion mutants were in trans complemented with the broad-host range expression vector pBBR1MCS-4ldi and pBBR1MCS-2geoA, restoring in both cases the wild type phenotype. Conclusions In-frame deletion mutants of genes in the anaerobic β-myrcene degradation revealed novel insights in the in vivo function. The deletion of a high-affinity geraniol dehydrogenase hampered, but did not preclude growth on monoterpenes. A second geraniol dehydrogenase activity was present that contributes to the β-myrcene degradation pathway. Growth on cyclic monoterpenes

  18. PHOSPHOLIPIDS OF FIVE PSEUDOMONAD ARCHETYPES FOR DIFFERENT TOLUENE DEGRADATION PATHWAYS

    Science.gov (United States)

    Liquid chromatography/electrospray ionization/mass spectrometry (LC/ESI/MS) was used to determine phospholipid profiles for five reference pseudomonad strains harboring distinct toluene catabolic pathways: Pseudomonas putida mt-2, Pseudomonas putida F1, Burkholderia cepacia G4, B...

  19. Metabolic pathways for degradation of aromatic hydrocarbons by bacteria

    NARCIS (Netherlands)

    Ladino-Orjuela, G.; Gomes, E.; da Silva, R.; Salt, C.; Parsons, J.R.; de Voogt, W.P.

    2016-01-01

    The aim of this revision was to build an updated collection of information focused on the mechanisms and elements involved in metabolic pathways of aromatic hydrocarbons by bacteria. Enzymes as an expression of the genetic load and the type of electron acceptor available, as an environmental factor,

  20. Pathways for degradation of plastic polymers floating in the marine environment.

    Science.gov (United States)

    Gewert, Berit; Plassmann, Merle M; MacLeod, Matthew

    2015-09-01

    Each year vast amounts of plastic are produced worldwide. When released to the environment, plastics accumulate, and plastic debris in the world's oceans is of particular environmental concern. More than 60% of all floating debris in the oceans is plastic and amounts are increasing each year. Plastic polymers in the marine environment are exposed to sunlight, oxidants and physical stress, and over time they weather and degrade. The degradation processes and products must be understood to detect and evaluate potential environmental hazards. Some attention has been drawn to additives and persistent organic pollutants that sorb to the plastic surface, but so far the chemicals generated by degradation of the plastic polymers themselves have not been well studied from an environmental perspective. In this paper we review available information about the degradation pathways and chemicals that are formed by degradation of the six plastic types that are most widely used in Europe. We extrapolate that information to likely pathways and possible degradation products under environmental conditions found on the oceans' surface. The potential degradation pathways and products depend on the polymer type. UV-radiation and oxygen are the most important factors that initiate degradation of polymers with a carbon-carbon backbone, leading to chain scission. Smaller polymer fragments formed by chain scission are more susceptible to biodegradation and therefore abiotic degradation is expected to precede biodegradation. When heteroatoms are present in the main chain of a polymer, degradation proceeds by photo-oxidation, hydrolysis, and biodegradation. Degradation of plastic polymers can lead to low molecular weight polymer fragments, like monomers and oligomers, and formation of new end groups, especially carboxylic acids.

  1. 3'-5' RNA degradation pathways in human cells

    DEFF Research Database (Denmark)

    Lubas, Michal Szymon

    RNA synthesis and degradation are key steps in the regulation of gene expression in all living organisms. During the course of his PhD studies, Michal Lubas centred his research on the nuclear and cytoplasmic RNA turnover of both noncoding and coding RNAs in human cells. His proteomic studies...... of the cytoplasmic 3'-5' exoribonuclease hDIS3L2. Using low throughout and high throughput techniques, both in vivo and in vitro, he characterised the nuclease and disclosed the role of hDIS3L2 in cytoplasmic mRNA metabolism....

  2. Evolutions of microbial degradation pathways for parent xenobiotic and for its metabolites follow different schemes.

    Science.gov (United States)

    Chong, Nyuk-Min; Chang, Chun-Shuo; Tsai, Shiu-Ching

    2012-09-01

    The pathways used by microorganisms for the metabolism of every xenobiotic substrate are specific. The catabolism of a xenobiotic goes through a series of intermediate steps and lower intermediates (metabolites) appear in sequence. The structure of the metabolites can be similar to the parents due to kinship. The purposes of this study were to examine if the degradation pathways that were developed for a parent xenobiotic are effective to degrade the parent's lower metabolites, and if the reverse is true. The xenobiotic substrates, 2,4-dichlorophenoxyacetic acid (2,4-D, the parent xenobiotic) and its metabolite 2,4-dichlorophenol (2,4-DCP), were independently subjected to acclimation and degradation tests by the biomasses of mixed-culture activated sludge and a pure culture of Arthrobacter sp. Activated sludge and Arthrobacter sp. that were acclimated to 2,4-D effectively degraded 2,4-D and the lower metabolites of 2,4-D, typically 2,4-DCP. During the degradation of 2,4-D, accumulations of the lower metabolites of 2,4-D were not found. The degradation pathways acquired from acclimation to 2,4-D are effective for all the metabolites of 2,4-D. However, pathways acquired from acclimation to 2,4-DCP are not effective in the degradation of the parent 2,4-D. Microorganisms acclimated to 2,4-D evolve their degradation pathways by a scheme that is different from the scheme the microorganisms employ when they are acclimated to the metabolites of 2,4-D.

  3. Degradation of streptomycin in aquatic environment: kinetics, pathway, and antibacterial activity analysis.

    Science.gov (United States)

    Shen, Yanru; Zhao, Wenyan; Zhang, Chunling; Shan, Yujie; Shi, Junxian

    2017-06-01

    Streptomycin used in human and veterinary medicine is released into the environment mainly through excretions. As such, its elimination in water should be investigated to control pollution. In this study, the degradation of streptomycin in water was studied, and the influence of variables, including light exposure, solution pH, temperature, ionic strength, dissolved organic matter (DOM), and coexisting surfactants, on degradation was investigated. Streptomycin degradation was consistent with the first-order model in aquatic environments. Its degradation rate under light exposure was 2.6-fold faster than that in the dark. Streptomycin was stable under neutral conditions, but it was easily decomposed in acidic and basic environments. Streptomycin degradation was enhanced by high temperature, and its half-life decreased from 103.4 days at 15 °C to 30.9 days at 40 °C. This process was also accelerated by the presence of Ca(2+) and slightly improved by the addition of HA. Streptomycin degradation was suppressed by high levels of the cationic surfactant cetyltri- methylammonium bromide (CTAB), but was promoted by the anionic surfactant sodium dodecyl benzene sulfonate (SDBS). The main degradation intermediates/products were identified through liquid chromatography-mass spectrometry, and the possible degradation pathway was proposed. The antibacterial activity of streptomycin solution was also determined during degradation. Results showed that STR degradation generated intermediates/products with weaker antibacterial activity than the parent compound.

  4. Connecting lignin-degradation pathway with pretreatment inhibitor sensitivity of Cupriavidus necator

    Directory of Open Access Journals (Sweden)

    Wei eWang

    2014-05-01

    Full Text Available To produce lignocellulosic biofuels economically, the complete release of monomers from the plant cell wall components, cellulose, hemicellulose and lignin, through pretreatment and hydrolysis (both enzymatic and chemical, and the efficient utilization of these monomers as carbon sources, is crucial. In addition, the identification and development of robust microbial biofuel production strains that can tolerate the toxic compounds generated during pretreatment and hydrolysis is also essential. In this work, Cupriavidus necator was selected due to its capabilities for utilizing lignin monomers and producing polyhydroxylbutyrate (PHB, a bioplastic as well as an advanced biofuel intermediate. We characterized the growth kinetics of C. necator in pretreated corn stover slurry as well as individually in the presence of 11 potentially toxic compounds in the saccharified slurry. We found that C. necator was sensitive to the saccharified slurry produced from dilute acid pretreated corn stover. Five out of 11 compounds within the slurry were characterized as toxic to C. necator, namely ammonium acetate, furfural, hydroxymethylfurfural (HMF, benzoic acid, and p-coumaric acid. Aldehydes (e.g., furfural and HMF were more toxic than the acetate and the lignin degradation products benzoic acid and p-coumaric acid; furfural was identified as the most toxic compound. Although toxic to C. necator at high concentration, ammonium acetate, benzoic acid, and p-coumaric acid could be utilized by C. necator with a stimulating effect on C. necator growth. Consequently, the lignin degradation pathway of C. necator was reconstructed based on genomic information and literature. The efficient conversion of intermediate catechol to downstream products of cis,cis-muconate or 2-hydroxymuconate-6-semialdehyde may help improve the robustness of C. necator to benzoic acid and p-coumaric acid as well as improve PHB productivity.

  5. Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator

    Science.gov (United States)

    Wang, Wei; Yang, Shihui; Hunsinger, Glendon B.; Pienkos, Philip T.; Johnson, David K.

    2014-01-01

    To produce lignocellulosic biofuels economically, the complete release of monomers from the plant cell wall components, cellulose, hemicellulose, and lignin, through pre-treatment and hydrolysis (both enzymatic and chemical), and the efficient utilization of these monomers as carbon sources, is crucial. In addition, the identification and development of robust microbial biofuel production strains that can tolerate the toxic compounds generated during pre-treatment and hydrolysis is also essential. In this work, Cupriavidus necator was selected due to its capabilities for utilizing lignin monomers and producing polyhydroxylbutyrate (PHB), a bioplastic as well as an advanced biofuel intermediate. We characterized the growth kinetics of C. necator in pre-treated corn stover slurry as well as individually in the pre-sence of 11 potentially toxic compounds in the saccharified slurry. We found that C. necator was sensitive to the saccharified slurry produced from dilute acid pre-treated corn stover. Five out of 11 compounds within the slurry were characterized as toxic to C. necator, namely ammonium acetate, furfural, hydroxymethylfurfural (HMF), benzoic acid, and p-coumaric acid. Aldehydes (e.g., furfural and HMF) were more toxic than the acetate and the lignin degradation products benzoic acid and p-coumaric acid; furfural was identified as the most toxic compound. Although toxic to C. necator at high concentration, ammonium acetate, benzoic acid, and p-coumaric acid could be utilized by C. necator with a stimulating effect on C. necator growth. Consequently, the lignin degradation pathway of C. necator was reconstructed based on genomic information and literature. The efficient conversion of intermediate catechol to downstream products of cis,cis-muconate or 2-hydroxymuconate-6-semialdehyde may help improve the robustness of C. necator to benzoic acid and p-coumaric acid as well as improve PHB productivity. PMID:24904560

  6. Connecting Lignin-Degradation Pathway with Pre-Treatment Inhibitor Sensitivity of Cupriavidus necator

    Energy Technology Data Exchange (ETDEWEB)

    Wang, W. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Yang, S. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Hunsinger, G. B. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Pienkos, P. T. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Johnson, D. K. [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2014-05-27

    In order to produce lignocellulosic biofuels economically, the complete release of monomers from the plant cell wall components, cellulose, hemicellulose, and lignin, through pre-treatment and hydrolysis (both enzymatic and chemical), and the efficient utilization of these monomers as carbon sources, is crucial. In addition, the identification and development of robust microbial biofuel production strains that can tolerate the toxic compounds generated during pre-treatment and hydrolysis is also essential. In this work, Cupriavidus necator was selected due to its capabilities for utilizing lignin monomers and producing polyhydroxylbutyrate (PHB), a bioplastic as well as an advanced biofuel intermediate. We characterized the growth kinetics of C. necator in pre-treated corn stover slurry as well as individually in the pre-sence of 11 potentially toxic compounds in the saccharified slurry. We found that C. necator was sensitive to the saccharified slurry produced from dilute acid pre-treated corn stover. Five out of 11 compounds within the slurry were characterized as toxic to C. necator, namely ammonium acetate, furfural, hydroxymethylfurfural (HMF), benzoic acid, and p-coumaric acid. Aldehydes (e.g., furfural and HMF) were more toxic than the acetate and the lignin degradation products benzoic acid and p-coumaric acid; furfural was identified as the most toxic compound. Although toxic to C. necator at high concentration, ammonium acetate, benzoic acid, and p-coumaric acid could be utilized by C. necator with a stimulating effect on C. necator growth. Consequently, the lignin degradation pathway of C. necator was reconstructed based on genomic information and literature. The efficient conversion of intermediate catechol to downstream products of cis,cis-muconate or 2-hydroxymuconate-6-semialdehyde may help improve the robustness of C. necator to benzoic acid and p-coumaric acid as well as improve PHB productivity.

  7. Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator.

    Science.gov (United States)

    Wang, Wei; Yang, Shihui; Hunsinger, Glendon B; Pienkos, Philip T; Johnson, David K

    2014-01-01

    To produce lignocellulosic biofuels economically, the complete release of monomers from the plant cell wall components, cellulose, hemicellulose, and lignin, through pre-treatment and hydrolysis (both enzymatic and chemical), and the efficient utilization of these monomers as carbon sources, is crucial. In addition, the identification and development of robust microbial biofuel production strains that can tolerate the toxic compounds generated during pre-treatment and hydrolysis is also essential. In this work, Cupriavidus necator was selected due to its capabilities for utilizing lignin monomers and producing polyhydroxylbutyrate (PHB), a bioplastic as well as an advanced biofuel intermediate. We characterized the growth kinetics of C. necator in pre-treated corn stover slurry as well as individually in the pre-sence of 11 potentially toxic compounds in the saccharified slurry. We found that C. necator was sensitive to the saccharified slurry produced from dilute acid pre-treated corn stover. Five out of 11 compounds within the slurry were characterized as toxic to C. necator, namely ammonium acetate, furfural, hydroxymethylfurfural (HMF), benzoic acid, and p-coumaric acid. Aldehydes (e.g., furfural and HMF) were more toxic than the acetate and the lignin degradation products benzoic acid and p-coumaric acid; furfural was identified as the most toxic compound. Although toxic to C. necator at high concentration, ammonium acetate, benzoic acid, and p-coumaric acid could be utilized by C. necator with a stimulating effect on C. necator growth. Consequently, the lignin degradation pathway of C. necator was reconstructed based on genomic information and literature. The efficient conversion of intermediate catechol to downstream products of cis,cis-muconate or 2-hydroxymuconate-6-semialdehyde may help improve the robustness of C. necator to benzoic acid and p-coumaric acid as well as improve PHB productivity.

  8. Unveiling New Degradation Intermediates/Pathways from the Photocatalytic Degradation of Microcystin-LR

    Science.gov (United States)

    This study focuses on the identification of reaction intermediates formed during the photocatalytic degradation of the cyanotoxin microcystin-LR with immobilized TiO2 Tphotocatalysts at neutral pH. To differentiate between impurities already existing in the MC-LR stand...

  9. Rapid reversal of translational silencing: Emerging role of microRNA degradation pathways in neuronal plasticity.

    Science.gov (United States)

    Fu, Xiuping; Shah, Aparna; Baraban, Jay M

    2016-09-01

    As microRNAs silence translation, rapid reversal of this process has emerged as an attractive mechanism for driving de novo protein synthesis mediating neuronal plasticity. Herein, we summarize recent studies identifying neuronal stimuli that trigger rapid decreases in microRNA levels and reverse translational silencing of plasticity transcripts. Although these findings indicate that neuronal stimulation elicits rapid degradation of selected microRNAs, we are only beginning to decipher the molecular pathways involved. Accordingly, we present an overview of several molecular pathways implicated in mediating microRNA degradation: Lin-28, translin/trax, and MCPIP1. As these degradation pathways target distinct subsets of microRNAs, they enable neurons to reverse silencing rapidly, yet selectively. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

    Institute of Scientific and Technical Information of China (English)

    QI Yun; ZHAO Lin; OJEKUNLE Z.Olusheyi; TAN Xin

    2007-01-01

    A study Was conducted to compare the diversity of 2-,3-,and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge.These samples contained strikingly different populations of mono-chlorobenzoate degraders.Although fewer cultures were isolated in the uncontaminated soils than contaminated one,the ability of microbial populations to mineralize chlorobenzoate was widespread.The 3-and 4-chlorobenzoate degraders were more diverse than me 2-chlorobenzoate degraders.One of the strains isolated from the sewage sludge was obtained.Based on its phenotype.chemotaxonomic properties and 16S rRNA gene,the organism S-7was classified as Rhodococcus erythropolis.The strain can grow at temperature from 4 to 37℃.C.It can utilize several(halo)aromatic compounds.Moreover,strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions.The psychrotolerant ability was significant for bioremediation in low temperature regions.Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain,but no(chloro)catechol 2,3-dioxygenase activities was detected.Spectral conversion assays with extracts from R.erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate.On the basis of these results,we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.

  11. RNAi screening for characterisation of ER-associated degradation pathways in mammalian cells

    DEFF Research Database (Denmark)

    Månsson, Mats David Joakim

    It is estimated that one third of all synthesized proteins in mammalian cells traverse the secretory pathway. Folding of proteins in the ER on their way to secretion is highly regulated. Proteins that are unable to achieve their native conformation are degraded by the ubiquitin-proteasome system...... fluorescence-based RNAi screens in mammalian cells on TCR-α-GFP and HANSκLC, for identification of ERAD pathways. By validating the obtained screening hits we concluded that UBE2J2 is involved in TCR-α-GFP degradation, possibly by ubiquitination of C-terminal serine residues in TCR-α-GFP. Additionally, we also...

  12. Elucidation of pathways of ribosomal RNA degradation: an essential role for RNase E

    Science.gov (United States)

    Sulthana, Shaheen; Basturea, Georgeta N.; Deutscher, Murray P.

    2016-01-01

    Although normally stable in growing cells, ribosomal RNAs are degraded under conditions of stress, such as starvation, and in response to misassembled or otherwise defective ribosomes in a process termed RNA quality control. Previously, our laboratory found that large fragments of 16S and 23S rRNA accumulate in strains lacking the processive exoribonucleases RNase II, RNase R, and PNPase, implicating these enzymes in the later steps of rRNA breakdown. Here, we define the pathways of rRNA degradation in the quality control process and during starvation, and show that the essential endoribonuclease, RNase E, is required to make the initial cleavages in both degradative processes. We also present evidence that explains why the exoribonuclease, RNase PH, is required to initiate the degradation of rRNA during starvation. The data presented here provide the first detailed description of rRNA degradation in bacterial cells. PMID:27298395

  13. Characterization of the transcriptome of Achromobacter sp. HZ01 with the outstanding hydrocarbon-degrading ability.

    Science.gov (United States)

    Hong, Yue-Hui; Deng, Mao-Cheng; Xu, Xiao-Ming; Wu, Chou-Fei; Xiao, Xi; Zhu, Qing; Sun, Xian-Xian; Zhou, Qian-Zhi; Peng, Juan; Yuan, Jian-Ping; Wang, Jiang-Hai

    2016-06-15

    Microbial remediation has become one of the most important strategies for eliminating petroleum pollutants. Revealing the transcript maps of microorganisms with the hydrocarbon-degrading ability contributes to enhance the degradation of hydrocarbons and further improve the effectiveness of bioremediation. In this study, we characterized the transcriptome of hydrocarbon-degrading Achromobacter sp. HZ01 after petroleum treatment for 16h. A total of 38,706,280 and 38,954,413 clean reads were obtained by RNA-seq for the petroleum-treated group and control, respectively. By an effective de novo assembly, 3597 unigenes were obtained, including 3485 annotated transcripts. Petroleum treatment had significantly influenced the transcriptional profile of strain HZ01, involving 742 differentially expressed genes. A part of genes were activated to exert specific physiological functions, whereas more genes were down-regulated including specific genes related to cell motility, genes associated with glycometabolism, and genes coding for ribosomal proteins. Identification of genes related to petroleum degradation revealed that the fatty acid metabolic pathway and a part of monooxygenases and dehydrogenases were activated, whereas the TCA cycle was inactive. Additionally, terminal oxidation might be a major aerobic pathway for the degradation of n-alkanes in strain HZ01. The newly obtained data contribute to better understand the gene expression profiles of hydrocarbon-degrading microorganisms after petroleum treatment, to further investigate the genetic characteristics of strain HZ01 and other related species and to develop cost-effective and eco-friendly strategies for remediation of crude oil-polluted environments.

  14. Terrestrial and marine perspectives on modeling organic matter degradation pathways.

    Science.gov (United States)

    Burd, Adrian B; Frey, Serita; Cabre, Anna; Ito, Takamitsu; Levine, Naomi M; Lønborg, Christian; Long, Matthew; Mauritz, Marguerite; Thomas, R Quinn; Stephens, Brandon M; Vanwalleghem, Tom; Zeng, Ning

    2016-01-01

    Organic matter (OM) plays a major role in both terrestrial and oceanic biogeochemical cycles. The amount of carbon stored in these systems is far greater than that of carbon dioxide (CO2 ) in the atmosphere, and annual fluxes of CO2 from these pools to the atmosphere exceed those from fossil fuel combustion. Understanding the processes that determine the fate of detrital material is important for predicting the effects that climate change will have on feedbacks to the global carbon cycle. However, Earth System Models (ESMs) typically utilize very simple formulations of processes affecting the mineralization and storage of detrital OM. Recent changes in our view of the nature of this material and the factors controlling its transformation have yet to find their way into models. In this review, we highlight the current understanding of the role and cycling of detrital OM in terrestrial and marine systems and examine how this pool of material is represented in ESMs. We include a discussion of the different mineralization pathways available as organic matter moves from soils, through inland waters to coastal systems and ultimately into open ocean environments. We argue that there is strong commonality between aspects of OM transformation in both terrestrial and marine systems and that our respective scientific communities would benefit from closer collaboration.

  15. Taxonomic characterization of the cellulose-degrading bacterium NCIB 10462

    Energy Technology Data Exchange (ETDEWEB)

    Dees, C.; Ringleberg, D.; Scott, T.C. [Oak Ridge National Lab., TN (United States); Phelps, T. [Univ. of Tennessee, Knoxville, TN (United States)

    1994-06-01

    The gram negative cellulase-producing bacterium NCIB 10462 has been previously named Pseudomonas fluorescens subsp. or var. cellulosa. Since there is renewed interest in cellulose-degrading bacteria for use in bioconversion of cellulose to chemical feed stocks and fuels, we re-examined the characteristics of this microorganism to determine its proper taxonomic characterization and to further define it`s true metabolic potential. Metabolic and physical characterization of NCIB 10462 revealed that this was an alkalophilic, non-fermentative, gram negative, oxidase positive, motile, cellulose-degrading bacterium. The aerobic substrate utilization profile of this bacterium was found to have few characteristics consistent with a classification of P. fluorescens with a very low probability match with the genus Sphingomonas. Total lipid analysis did not reveal that any sphingolipid bases are produced by this bacterium. NCIB 10462 was found to grow best aerobically but also grows well in complex media under reducing conditions. NCIB 10462 grew slowly under full anaerobic conditions on complex media but growth on cellulosic media was found only under aerobic conditions. Total fatty acid analysis (MIDI) of NCIB 10462 failed to group this bacterium with a known pseudomonas species. However, fatty acid analysis of the bacteria when grown at temperatures below 37{degrees}C suggest that the organism is a pseudomonad. Since a predominant characteristic of this bacterium is it`s ability to degrade cellulose, we suggest it be called Pseudomonas cellulosa.

  16. Stability-Indicating Method and LC-MS-MS Characterization of Forced Degradation Products of Sofosbuvir.

    Science.gov (United States)

    Nebsen, M; Elzanfaly, Eman S

    2016-07-19

    Sofosbuvir is a novel direct acting antiviral agent against hepatitis C virus. In the present work, a rapid, specific and reproducible isocratic reversed phase high performance liquid chromatography (RP-HPLC) method has been developed and validated for the determination of sofosbuvir in the presence of its stressed degradation products. Sobosbuvir was subjected to hydrolysis (acidic, alkaline and neutral), oxidation, photolysis and thermal stress, as per international conference on harmonization (ICH) conditions. The drug showed degradation under oxidative, photolysis, acid and base hydrolysis stress conditions. However, it was stable under thermal and neutral hydrolysis stress conditions. Chromatographic separation of the drug from its degradation products was performed on Inertsil ODS-3 C18 (250 mm × 4.6 mm i.d., 5 µm) column using a green mobile phase of methanol:water 70:30 (v/v). The degradation products were characterized by LC-MS-MS and the fragmentation pathways were proposed. The developed method was validated as per ICH guidelines. No previous method was reported regarding the degradation behavior of sofosbuvir.

  17. Degradation pathway of malachite green in a novel dual-tank photoelectrochemical catalytic reactor.

    Science.gov (United States)

    Diao, Zenghui; Li, Mingyu; Zeng, Fanyin; Song, Lin; Qiu, Rongliang

    2013-09-15

    A novel dual-tank photoelectrochemical catalytic reactor was designed to investigate the degradation pathway of malachite green. A thermally formed TiO₂/Ti thin film electrode was used as photoanode, graphite was used as cathode, and a saturated calomel electrode was employed as the reference electrode in the reactor. In the reactor, the anode and cathode tanks were connected by a cation exchange membrane. Results showed that the decolorization ratio of malachite green in the anode and cathode was 98.5 and 96.5% after 120 min, respectively. Malachite green in the two anode and cathode tanks was oxidized, achieving the bipolar double effect. Malachite green in both the anode and cathode tanks exhibited similar catalytic degradation pathways. The double bond of the malachite green molecule was attacked by strong oxidative hydroxyl radicals, after which the organic compound was degraded by the two pathways into 4,4-bis(dimethylamino) benzophenone, 4-(dimethylamino) benzophenone, 4-(dimethylamino) phenol, and other intermediate products. Eventually, malachite green was degraded into oxalic acid as a small molecular organic acid, which was degraded by processes such as demethylation, deamination, nitration, substitution, addition, and other reactions. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Degradation of oxcarbazepine by UV-activated persulfate oxidation: kinetics, mechanisms, and pathways.

    Science.gov (United States)

    Bu, Lingjun; Zhou, Shiqing; Shi, Zhou; Deng, Lin; Li, Guangchao; Yi, Qihang; Gao, Naiyun

    2016-02-01

    The degradation kinetics and mechanism of the antiepileptic drug oxcarbazepine (OXC) by UV-activated persulfate oxidation were investigated in this study. Results showed that UV/persulfate (UV/PS) process appeared to be more effective in degrading OXC than UV or PS alone. The OXC degradation exhibited a pseudo-first order kinetics pattern and the degradation rate constants (k obs) were affected by initial OXC concentration, PS dosage, initial pH, and humic acid concentration to different degrees. It was found that low initial OXC concentration, high persulfate dosage, and initial pH enhanced the OXC degradation. Additionally, the presence of humic acid in the solution could greatly inhibit the degradation of OXC. Moreover, hydroxyl radical (OH•) and sulfate radical (SO4 (-)••) were identified to be responsible for OXC degradation and SO4 (-)• made the predominant contribution in this study. Finally, major intermediate products were identified and a preliminary degradation pathway was proposed. Results demonstrated that UV/PS system is a potential technology to control the water pollution caused by emerging contaminants such as OXC.

  19. A second pathway to degrade pyrimidine nucleic acid precursors in eukaryotes

    DEFF Research Database (Denmark)

    Andersen, Gorm; Bjornberg, Olof; Polakova, Silvia;

    2008-01-01

    Pyrimidine bases are the central precursors for RNA and DNA, and their intracellular pools are determined by de novo, salvage and catabolic pathways. In eukaryotes, degradation of uracil has been believed to proceed only via the reduction to dihydrouracil. Using a yeast model, Saccharomyces kluyv...

  20. Kinetic models and pathways of ronidazole degradation by chlorination, UV irradiation and UV/chlorine processes.

    Science.gov (United States)

    Qin, Lang; Lin, Yi-Li; Xu, Bin; Hu, Chen-Yan; Tian, Fu-Xiang; Zhang, Tian-Yang; Zhu, Wen-Qian; Huang, He; Gao, Nai-Yun

    2014-11-15

    Degradation kinetics and pathways of ronidazole (RNZ) by chlorination (Cl2), UV irradiation and combined UV/chlorine processes were investigated in this paper. The degradation kinetics of RNZ chlorination followed a second-order behavior with the rate constants calculated as (2.13 ± 0.15) × 10(2) M(-2) s(-1), (0.82 ± 0.52) × 10(-2) M(-1) s(-1) and (2.06 ± 0.09) × 10(-1) M(-1) s(-1) for the acid-catalyzed reaction, as well as the reactions of RNZ with HOCl and OCl(-), respectively. Although UV irradiation degraded RNZ more effectively than chlorination did, very low quantum yield of RNZ at 254 nm was obtained as 1.02 × 10(-3) mol E(-1). RNZ could be efficiently degraded and mineralized in the UV/chlorine process due to the generation of hydroxyl radicals. The second-order rate constant between RNZ and hydroxyl radical was determined as (2.92 ± 0.05) × 10(9) M(-1) s(-1). The degradation intermediates of RNZ during the three processes were identified with Ultra Performance Liquid Chromatography - Electrospray Ionization - mass spectrometry and the degradation pathways were then proposed. Moreover, the variation of chloropicrin (TCNM) and chloroform (CF) formation after the three processes were further evaluated. Enhanced formation of CF and TCNM precursors during UV/chlorine process deserves extensive attention in drinking water treatment.

  1. Evolutionary, computational, and biochemical studies of the salicylaldehyde dehydrogenases in the naphthalene degradation pathway

    Science.gov (United States)

    Jia, Baolei; Jia, Xiaomeng; Hyun Kim, Kyung; Ji Pu, Zhong; Kang, Myung-Suk; Ok Jeon, Che

    2017-01-01

    Salicylaldehyde (SAL) dehydrogenase (SALD) is responsible for the oxidation of SAL to salicylate using nicotinamide adenine dinucleotide (NAD+) as a cofactor in the naphthalene degradation pathway. We report the use of a protein sequence similarity network to make functional inferences about SALDs. Network and phylogenetic analyses indicated that SALDs and the homologues are present in bacteria and fungi. The key residues in SALDs were analyzed by evolutionary methods and a molecular simulation analysis. The results showed that the catalytic residue is most highly conserved, followed by the residues binding NAD+ and then the residues binding SAL. A molecular simulation analysis demonstrated the binding energies of the amino acids to NAD+ and/or SAL and showed that a conformational change is induced by binding. A SALD from Alteromonas naphthalenivorans (SALDan) that undergoes trimeric oligomerization was characterized enzymatically. The results showed that SALDan could catalyze the oxidation of a variety of aromatic aldehydes. Site-directed mutagenesis of selected residues binding NAD+ and/or SAL affected the enzyme’s catalytic efficiency, but did not eliminate catalysis. Finally, the relationships among the evolution, catalytic mechanism, and functions of SALD are discussed. Taken together, this study provides an expanded understanding of the evolution, functions, and catalytic mechanism of SALD. PMID:28233868

  2. Modelling pathways to Rubisco degradation: a structural equation network modelling approach.

    Directory of Open Access Journals (Sweden)

    Catherine Tétard-Jones

    Full Text Available 'Omics analysis (transcriptomics, proteomics quantifies changes in gene/protein expression, providing a snapshot of changes in biochemical pathways over time. Although tools such as modelling that are needed to investigate the relationships between genes/proteins already exist, they are rarely utilised. We consider the potential for using Structural Equation Modelling to investigate protein-protein interactions in a proposed Rubisco protein degradation pathway using previously published data from 2D electrophoresis and mass spectrometry proteome analysis. These informed the development of a prior model that hypothesised a pathway of Rubisco Large Subunit and Small Subunit degradation, producing both primary and secondary degradation products. While some of the putative pathways were confirmed by the modelling approach, the model also demonstrated features that had not been originally hypothesised. We used Bayesian analysis based on Markov Chain Monte Carlo simulation to generate output statistics suggesting that the model had replicated the variation in the observed data due to protein-protein interactions. This study represents an early step in the development of approaches that seek to enable the full utilisation of information regarding the dynamics of biochemical pathways contained within proteomics data. As these approaches gain attention, they will guide the design and conduct of experiments that enable 'Omics modelling to become a common place practice within molecular biology.

  3. New hydrocarbon degradation pathways in the microbial metagenome from Brazilian petroleum reservoirs.

    Directory of Open Access Journals (Sweden)

    Isabel Natalia Sierra-García

    Full Text Available Current knowledge of the microbial diversity and metabolic pathways involved in hydrocarbon degradation in petroleum reservoirs is still limited, mostly due to the difficulty in recovering the complex community from such an extreme environment. Metagenomics is a valuable tool to investigate the genetic and functional diversity of previously uncultured microorganisms in natural environments. Using a function-driven metagenomic approach, we investigated the metabolic abilities of microbial communities in oil reservoirs. Here, we describe novel functional metabolic pathways involved in the biodegradation of aromatic compounds in a metagenomic library obtained from an oil reservoir. Although many of the deduced proteins shared homology with known enzymes of different well-described aerobic and anaerobic catabolic pathways, the metagenomic fragments did not contain the complete clusters known to be involved in hydrocarbon degradation. Instead, the metagenomic fragments comprised genes belonging to different pathways, showing novel gene arrangements. These results reinforce the potential of the metagenomic approach for the identification and elucidation of new genes and pathways in poorly studied environments and contribute to a broader perspective on the hydrocarbon degradation processes in petroleum reservoirs.

  4. New Hydrocarbon Degradation Pathways in the Microbial Metagenome from Brazilian Petroleum Reservoirs

    Science.gov (United States)

    Sierra-García, Isabel Natalia; Correa Alvarez, Javier; Pantaroto de Vasconcellos, Suzan; Pereira de Souza, Anete; dos Santos Neto, Eugenio Vaz; de Oliveira, Valéria Maia

    2014-01-01

    Current knowledge of the microbial diversity and metabolic pathways involved in hydrocarbon degradation in petroleum reservoirs is still limited, mostly due to the difficulty in recovering the complex community from such an extreme environment. Metagenomics is a valuable tool to investigate the genetic and functional diversity of previously uncultured microorganisms in natural environments. Using a function-driven metagenomic approach, we investigated the metabolic abilities of microbial communities in oil reservoirs. Here, we describe novel functional metabolic pathways involved in the biodegradation of aromatic compounds in a metagenomic library obtained from an oil reservoir. Although many of the deduced proteins shared homology with known enzymes of different well-described aerobic and anaerobic catabolic pathways, the metagenomic fragments did not contain the complete clusters known to be involved in hydrocarbon degradation. Instead, the metagenomic fragments comprised genes belonging to different pathways, showing novel gene arrangements. These results reinforce the potential of the metagenomic approach for the identification and elucidation of new genes and pathways in poorly studied environments and contribute to a broader perspective on the hydrocarbon degradation processes in petroleum reservoirs. PMID:24587220

  5. Tissue factor pathway inhibitor relates to fibrin degradation in patients with acute deep venous thrombosis

    DEFF Research Database (Denmark)

    Sidelmann, Johannes J; Bladbjerg, Else-Marie; Gram, Jørgen

    2008-01-01

    Reduced concentration of tissue factor pathway inhibitor is a risk factor for development of deep venous thrombosis, whereas elevated concentrations of tissue factor pathway inhibitor are observed in patients with acute myocardial infarction and disseminated intravascular coagulation. Presently, we...... studied the association between inflammation, endothelial cell perturbation, fibrin degradation and the concentration of tissue factor pathway inhibitor in patients suspected for acute deep venous thrombosis. We determined the tissue factor pathway inhibitor -33T/C polymorphism, free and total tissue...... factor pathway inhibitor, C-reactive protein, von Willebrand factor and D-Dimer in 160 consecutive patients admitted to hospital with a tentative diagnosis of acute deep venous thrombosis. Deep venous thrombosis was identified in 57 patients (18 distal and 39 proximal). The distribution of the tissue...

  6. Bacterial community structure and predicted alginate metabolic pathway in an alginate-degrading bacterial consortium.

    Science.gov (United States)

    Kita, Akihisa; Miura, Toyokazu; Kawata, Satoshi; Yamaguchi, Takeshi; Okamura, Yoshiko; Aki, Tsunehiro; Matsumura, Yukihiko; Tajima, Takahisa; Kato, Junichi; Nishio, Naomichi; Nakashimada, Yutaka

    2016-03-01

    Methane fermentation is one of the effective approaches for utilization of brown algae; however, this process is limited by the microbial capability to degrade alginate, a main polysaccharide found in these algae. Despite its potential, little is known about anaerobic microbial degradation of alginate. Here we constructed a bacterial consortium able to anaerobically degrade alginate. Taxonomic classification of 16S rRNA gene, based on high-throughput sequencing data, revealed that this consortium included two dominant strains, designated HUA-1 and HUA-2; these strains were related to Clostridiaceae bacterium SK082 (99%) and Dysgonomonas capnocytophagoides (95%), respectively. Alginate lyase activity and metagenomic analyses, based on high-throughput sequencing data, revealed that this bacterial consortium possessed putative genes related to a predicted alginate metabolic pathway. However, HUA-1 and 2 did not grow on agar medium with alginate by using roll-tube method, suggesting the existence of bacterial interactions like symbiosis for anaerobic alginate degradation.

  7. LC, MSn and LC-MS/MS studies for the characterization of degradation products of amlodipine

    Institute of Scientific and Technical Information of China (English)

    Ravi N. Tiwari; Nishit Shah; Vikas Bhalani; Anand Mahajan

    2015-01-01

    In the present study, comprehensive stress testing of amlodipine (AM) was carried out according to International Conference on Harmonization (ICH) Q1A(R2) guideline. AM was subjected to acidic, neutral and alkaline hydrolysis, oxidation, photolysis and thermal stress conditions. The drug showed instability in acidic and alkaline conditions, while it remained stable to neutral, oxidative, light and thermal stress. A total of nine degradation products (DPs) were formed from AM, which could be separated by the developed gradient LC method on a C18 column. The products formed under various stress conditions were investigated by LC–MS/MS analysis. The previously developed LC method was suitably modified for LC–MS/MS studies by replacing phosphate buffer with ammonium acetate buffer of the same concentration (pH 5.0). A complete fragmentation pathway of the drug was first established to characterize all the degradation products using LC–MS/MS and multi-stage mass (MSn) fragmentation studies. The obtained mass values were used to study elemental compositions, and the total information helped with the identification of DPs, along with its degradation pathway.

  8. LC, MSn and LC–MS/MS studies for the characterization of degradation products of amlodipine

    Directory of Open Access Journals (Sweden)

    Ravi N. Tiwari

    2015-02-01

    Full Text Available In the present study, comprehensive stress testing of amlodipine (AM was carried out according to International Conference on Harmonization (ICH Q1A(R2 guideline. AM was subjected to acidic, neutral and alkaline hydrolysis, oxidation, photolysis and thermal stress conditions. The drug showed instability in acidic and alkaline conditions, while it remained stable to neutral, oxidative, light and thermal stress. A total of nine degradation products (DPs were formed from AM, which could be separated by the developed gradient LC method on a C18 column. The products formed under various stress conditions were investigated by LC–MS/MS analysis. The previously developed LC method was suitably modified for LC–MS/MS studies by replacing phosphate buffer with ammonium acetate buffer of the same concentration (pH 5.0. A complete fragmentation pathway of the drug was first established to characterize all the degradation products using LC–MS/MS and multi-stage mass (MSn fragmentation studies. The obtained mass values were used to study elemental compositions, and the total information helped with the identification of DPs, along with its degradation pathway.

  9. Degradation kinetics and pathway of phenol by Pseudomonas and Bacillus species.

    Science.gov (United States)

    Hasan, Syed Adnan; Jabeen, Suraiya

    2015-01-02

    This article elucidates that strain Pseudomonas aeruginosa (IES-Ps-1) is a versatile toxic organic compound degrader. With the degradation of malathion and cypermethrin (studied by other researchers previously), this strain was able to degrade phenol. Two other indigenous soil flora (i.e., Pseudomonas sp. (IES-S) and Bacillus subtilis (IES-B)) were also found to be potential phenol degraders. Phenol was degraded with Monod kinetics during growth in nutrient broth and mineral salts medium. Before entering into the growth inhibition phase, strains IES-Ps-1, IES-S and IES-B could tolerate up to 400, 700 and 500 mg/L phenol, respectively, when contained in nutrient broth. However, according to the Luong-Levenspiel model, the growth of strains IES-Ps-1, IES-S and IES-B would cease at 2000, 2174 and 2190 mg/L phenol, respectively. Strain IES-Ps-1 degraded 700, 900 and 1050 mg/L phenol contained in mineral salts medium with the specific rates of 0.034, 0.075 and 0.021 h(-1), respectively. All these strains grew by making clusters when exposed to phenol in order to prevent damages due to high substrate concentration. These strains transformed phenol into catechol, which was then degraded via ortho-cleavage pathway.

  10. Nuclear mRNA degradation pathway(s are implicated in Xist regulation and X chromosome inactivation.

    Directory of Open Access Journals (Sweden)

    Constance Ciaudo

    2006-06-01

    Full Text Available A critical step in X-chromosome inactivation (XCI, which results in the dosage compensation of X-linked gene expression in mammals, is the coating of the presumptive inactive X chromosome by the large noncoding Xist RNA, which then leads to the recruitment of other factors essential for the heterochromatinisation of the inactive X and its transcriptional silencing. In an approach aimed at identifying genes implicated in the X-inactivation process by comparative transcriptional profiling of female and male mouse gastrula, we identified the Eif1 gene involved in translation initiation and RNA degradation. We show here that female embryonic stem cell lines, silenced by RNA interference for the Eif1 gene, are unable to form Xist RNA domains upon differentiation and fail to undergo X-inactivation. To probe further an effect involving RNA degradation pathways, the inhibition by RNA interference of Rent1, a factor essential for nonsense-mediated decay and Exosc10, a specific nuclear component of the exosome, was analysed and shown to similarly impair Xist upregulation and XCI. In Eif1-, Rent1-, and Exosc10-interfered clones, Xist spliced form(s are strongly downregulated, while the levels of unspliced form(s of Xist and the stability of Xist RNA remain comparable to that of the control cell lines. Our data suggests a role for mRNA nuclear degradation pathways in the critical regulation of spliced Xist mRNA levels and the onset of the X-inactivation process.

  11. Identification of the para-nitrophenol catabolic pathway, and characterization of three enzymes involved in the hydroquinone pathway, in pseudomonas sp. 1-7

    Directory of Open Access Journals (Sweden)

    Zhang Shuangyu

    2012-03-01

    Full Text Available Abstract Background para-Nitrophenol (PNP, a priority environmental pollutant, is hazardous to humans and animals. However, the information relating to the PNP degradation pathways and their enzymes remain limited. Results Pseudomonas sp.1-7 was isolated from methyl parathion (MP-polluted activated sludge and was shown to degrade PNP. Two different intermediates, hydroquinone (HQ and 4-nitrocatechol (4-NC were detected in the catabolism of PNP. This indicated that Pseudomonas sp.1-7 degraded PNP by two different pathways, namely the HQ pathway, and the hydroxyquinol (BT pathway (also referred to as the 4-NC pathway. A gene cluster (pdcEDGFCBA was identified in a 10.6 kb DNA fragment of a fosmid library, which cluster encoded the following enzymes involved in PNP degradation: PNP 4-monooxygenase (PdcA, p-benzoquinone (BQ reductase (PdcB, hydroxyquinol (BT 1,2-dioxygenase (PdcC, maleylacetate (MA reductase (PdcF, 4-hydroxymuconic semialdehyde (4-HS dehydrogenase (PdcG, and hydroquinone (HQ 1,2-dioxygenase (PdcDE. Four genes (pdcDEFG were expressed in E. coli and the purified pdcDE, pdcG and pdcF gene products were shown to convert HQ to 4-HS, 4-HS to MA and MA to β-ketoadipate respectively by in vitro activity assays. Conclusions The cloning, sequencing, and characterization of these genes along with the functional PNP degradation studies identified 4-NC, HQ, 4-HS, and MA as intermediates in the degradation pathway of PNP by Pseudomonas sp.1-7. This is the first conclusive report for both 4-NC and HQ- mediated degradation of PNP by one microorganism.

  12. Metagenomic identification of bacterioplankton taxa and pathways involved in microcystin degradation in lake erie.

    Directory of Open Access Journals (Sweden)

    Xiaozhen Mou

    Full Text Available Cyanobacterial harmful blooms (CyanoHABs that produce microcystins are appearing in an increasing number of freshwater ecosystems worldwide, damaging quality of water for use by human and aquatic life. Heterotrophic bacteria assemblages are thought to be important in transforming and detoxifying microcystins in natural environments. However, little is known about their taxonomic composition or pathways involved in the process. To address this knowledge gap, we compared the metagenomes of Lake Erie free-living bacterioplankton assemblages in laboratory microcosms amended with microcystins relative to unamended controls. A diverse array of bacterial phyla were responsive to elevated supply of microcystins, including Acidobacteria, Actinobacteria, Bacteroidetes, Planctomycetes, Proteobacteria of the alpha, beta, gamma, delta and epsilon subdivisions and Verrucomicrobia. At more detailed taxonomic levels, Methylophilales (mainly in genus Methylotenera and Burkholderiales (mainly in genera Bordetella, Burkholderia, Cupriavidus, Polaromonas, Ralstonia, Polynucleobacter and Variovorax of Betaproteobacteria were suggested to be more important in microcystin degradation than Sphingomonadales of Alphaproteobacteria. The latter taxa were previously thought to be major microcystin degraders. Homologs to known microcystin-degrading genes (mlr were not overrepresented in microcystin-amended metagenomes, indicating that Lake Erie bacterioplankton might employ alternative genes and/or pathways in microcystin degradation. Genes for xenobiotic metabolism were overrepresented in microcystin-amended microcosms, suggesting they are important in bacterial degradation of microcystin, a phenomenon that has been identified previously only in eukaryotic systems.

  13. Degradation kinetics and pathways of three calcium channel blockers under UV irradiation.

    Science.gov (United States)

    Zhu, Bing; Zonja, Bozo; Gonzalez, Oscar; Sans, Carme; Pérez, Sandra; Barceló, Damia; Esplugas, Santiago; Xu, Ke; Qiang, Zhimin

    2015-12-01

    Calcium channel blockers (CCBs) are a group of pharmaceuticals widely prescribed to lower blood pressure and treat heart diseases. They have been frequently detected in wastewater treatment plant (WWTP) effluents and downstream river waters, thus inducing a potential risk to aquatic ecosystems. However, little is known about the behavior and fate of CCBs under UV irradiation, which has been adopted as a primary disinfection method for WWTP effluents. This study investigated the degradation kinetics and pathways of three commonly-used CCBs, including amlodipine (AML), diltiazem (DIL), and verapamil (VER), under UV (254 nm) irradiation. The chemical structures of transformation byproducts (TBPs) were first identified to assess the potential ecological hazards. On that basis, a generic solid-phase extraction method, which simultaneously used four different cartridges, was adopted to extract and enrich the TBPs. Thereafter, the photo-degradation of target CCBs was performed under UV fluences typical for WWTP effluent disinfection. The degradation of all three CCBs conformed to the pseudo-first-order kinetics, with rate constants of 0.031, 0.044 and 0.011 min(-1) for AML, DIL and VER, respectively. By comparing the MS(2) fragments and the evolution (i.e., formation or decay) trends of identified TBPs, the degradation pathways were proposed. In the WWTP effluent, although the target CCBs could be degraded, several TBPs still contained the functional pharmacophores and reached peak concentrations under UV fluences of 40-100 mJ cm(-2).

  14. Metagenomic identification of bacterioplankton taxa and pathways involved in microcystin degradation in lake erie.

    Science.gov (United States)

    Mou, Xiaozhen; Lu, Xinxin; Jacob, Jisha; Sun, Shulei; Heath, Robert

    2013-01-01

    Cyanobacterial harmful blooms (CyanoHABs) that produce microcystins are appearing in an increasing number of freshwater ecosystems worldwide, damaging quality of water for use by human and aquatic life. Heterotrophic bacteria assemblages are thought to be important in transforming and detoxifying microcystins in natural environments. However, little is known about their taxonomic composition or pathways involved in the process. To address this knowledge gap, we compared the metagenomes of Lake Erie free-living bacterioplankton assemblages in laboratory microcosms amended with microcystins relative to unamended controls. A diverse array of bacterial phyla were responsive to elevated supply of microcystins, including Acidobacteria, Actinobacteria, Bacteroidetes, Planctomycetes, Proteobacteria of the alpha, beta, gamma, delta and epsilon subdivisions and Verrucomicrobia. At more detailed taxonomic levels, Methylophilales (mainly in genus Methylotenera) and Burkholderiales (mainly in genera Bordetella, Burkholderia, Cupriavidus, Polaromonas, Ralstonia, Polynucleobacter and Variovorax) of Betaproteobacteria were suggested to be more important in microcystin degradation than Sphingomonadales of Alphaproteobacteria. The latter taxa were previously thought to be major microcystin degraders. Homologs to known microcystin-degrading genes (mlr) were not overrepresented in microcystin-amended metagenomes, indicating that Lake Erie bacterioplankton might employ alternative genes and/or pathways in microcystin degradation. Genes for xenobiotic metabolism were overrepresented in microcystin-amended microcosms, suggesting they are important in bacterial degradation of microcystin, a phenomenon that has been identified previously only in eukaryotic systems.

  15. Kinetics and pathways of cyanide degradation at high temperatures and pressures.

    Science.gov (United States)

    Oulego, Paula; Laca, Adriana; Diaz, Mario

    2013-02-05

    The degradation of cyanide was performed in a 1-L semibatch reactor at temperatures between 393 and 473 K and at total pressures in the range of 2.0-8.0 MPa. The initial pH of the solution was set at 11, whereas initial concentrations ranged from 3.85 to 25 mM, which resemble the typical concentrations of cyanide-containing wastewater. The change with time of cyanide concentration, intermediates, and final products was analyzed in order to elucidate the reaction pathways. The experimental results suggest two parallel pathways of alkaline hydrolysis for the degradation of the pollutant. Formate and ammonia were identified as the final reaction products for one of the pathways, whereas carbon dioxide, nitrogen, and hydrogen were considered to be the final products for the other one. The degradation reaction results were fitted to first-order kinetic equations with respect to cyanide, giving respectively activation energies of 108.2 ± 3.3 and 77.6 ± 3.0 kJ/mol. Consequently, the formation of formate and ammonia is favored at high temperatures, whereas low temperatures favored the pathway leading to the formation of carbon dioxide and nitrogen.

  16. NIR is degraded by the anaphase-promoting complex proteasome pathway

    Directory of Open Access Journals (Sweden)

    Jeong Ho Myong

    2014-01-01

    Full Text Available Novel INHAT Repressor (NIR is a histone acetylation inhibitor that can directly bind histone complexes and the tumor suppressors p53 and p63. Because NIR is mainly localized in the nucleolus and disappears from the nucleolus upon RNase treatment, it is thought to bind RNA or ribonucleoproteins. When NIR moves to the cytoplasm, it is immediately degraded; this degradation was blocked by MG132, a proteasome inhibitor. Furthermore, the central domain of NIR specifically bound APC-CCdh1. These data show that the stability of NIR is governed by the ubiquitin/proteasome pathway.

  17. Isolation, Identification and Phenotypic Characterization of Microcystin-Degrading Bacteria from Lake Erie

    Science.gov (United States)

    Krishnan, A.; Mou, X. J.

    2015-12-01

    Lake Erie, the smallest and warmest lake among the Laurentian Great Lakes, is known for its problem of eutrophication and frequent occurrence of harmful cyanobacterial blooms (CyanoHABs). One major harmful effect of CyanoHABs is the production of cyanotoxins, especially microcystins. Microcystins (MC) are a group of hepatotoxins and the predominant variant of them is MC-LR. Field measurements and lab experiments indicate that MC degradation in Lake Erie is mainly carried out by indigenous bacteria. However, our knowledge on taxa involved in this process is very limited. This study aimed to fill this knowledge gap using a culture-dependent approach. Water and surface sediment samples were collected from Lake Erie in 2014 and 2015 and enriched with MC-LR. Cells were plated on a number of culturing media. The obtained pure bacterial cultures were screened for MC degrading abilities by MT2 BIO-LOG assays and by growing cells in liquid media containing MC-LR as the sole carbon source. In the latter experiment, MC concentrations were measured using HPLC. Isolates showing positive MC degradation activities in the screening steps were designated MC+ bacteria and characterized based on their phenotypic properties, including colony pigmentation, elevation, opacity, margin, gram nature and motility. The taxonomic identity of MC+ bacteria was determined by 16S rRNA gene full-length DNA sequencing. The presence of mlrA, a gene encoding MC cleavage pathway, was detected by PCR. Our culturing efforts obtained 520 pure cultures; 44 of them were identified as MC+. These MC+ isolates showed diversity in taxonomic identities and differed in their morphology, gram nature, colony characteristics and motility. PCR amplification of mlrA gene yield negative results for all MC+ isolates, indicating that the primers that were used may not be ubiquitous enough to cover the heterogeneity of mlrA genes or, more likely, alternative degradative genes/pathways were employed by Lake Erie bacteria

  18. [Degradation of L-phenylalanine and of aromatic carboxylic acids by chloridazon-degrading bacteria. Combination of side chain degradation and dioxygenase pathway].

    Science.gov (United States)

    Wegst, W; Lingens, F

    1981-09-01

    Strain N of Chloridazon-degrading bacteria degrades phenylalanine via cis-2,3-dihydro-2,3-dihydroxyphenylalanine,2,3-dihydroxyphenylalanine aspartate and 4-hydroxy-2-oxovalerate [Hoppe-Seyler's Z. Physiol. Chem. 360, 957--969, (1979); Biochem. J. 194, 679--684 (1981)]. cis-2,3-Dihydro-2,3-dihydroxyphenylalanine and 2,3-dihydroxyphenylalanine as well as phenylpyruvate, cis-2,3-dihydro-2,3-dihydroxyphenylpyruvate, 2,3-dihydroxyphenylpyruvate, cis-2,3-dihydro-2,3-dihydroxyphenylacetate, 2,3-dihydroxyphenylacetate and 2,3-dihydroxybenzaldehyde are detectable in the medium of strain E during growth on phenylalanine. Incubation with phenylacetate, 3-phenylpropionate or 4-phenylbutyrate leads to the accumulation of the corresponding cis-2,3-dihydro-2,3-dihydroxyphenyl derivatives. These compounds are transformed with dihydrodiol dehydrogenase to 2,3-dihydroxyphenylacetate, 3-(2,3-dihydroxyphenyl)propionate and 4-(2,3-dihydroxyphenyl)-butyrate, 3-(2,3-dihydroxyphenyl)propionate is attacked by a catechol 2,3-dioxygenase and the meta-cleavage product is again cleaved by a hydrolase yielding succinate. In a similar reaction sequence the degradation of 4-phenylbutyrate leads to the formation of glutarate. From the growth medium of strain E on phenylacetate also small amounts of 2-, 3- and 4-hydroxyphenylacetate were isolated. Resting cells were shown to metabolize 3- and 4-hydroxyphenylacetate via homogentisate and 3,4-dihydroxyphenylacetate. In the culture medium of strain K2AP benzoate could be detected. Pathways for the degradation of phenylalanine and aromatic carboxylic acids in chloridazon degrading bacteria are proposed.

  19. Degradation of 4-nitrocatechol by Burkholderia cepacia: a plasmid-encoded novel pathway.

    Science.gov (United States)

    Chauhan, A; Samanta, S K; Jain, R K

    2000-05-01

    Pseudomonas cepacia RKJ200 (now described as Burkholderia cepacia) has been shown to utilize p-nitrophenol (PNP) as sole carbon and energy source. The present work demonstrates that RKJ200 utilizes 4-nitrocatechol (NC) as the sole source of carbon, nitrogen and energy, and is degraded with concomitant release of nitrite ions. Several lines of evidence, including thin layer chromatography, gas chromatography, 1H-nuclear magnetic resonance, gas chromatography-mass spectrometry, spectral analyses and quantification of intermediates by high performance liquid chromatography, have shown that NC is degraded via 1,2, 4-benzenetriol (BT) and hydroquinone (HQ) formation. Studies carried out on a PNP- derivative and a PNP+ transconjugant also demonstrate that the genes for the NC degradative pathway reside on the plasmid present in RKJ200; the same plasmid had earlier been shown to encode genes for PNP degradation, which is also degraded via HQ formation. It is likely, therefore, that the same sets of genes encode the further metabolism of HQ in NC and PNP degradation.

  20. New metabolic pathway for degradation of 2-nitrobenzoate by Arthrobacter sp. SPG

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Arora

    2015-06-01

    Full Text Available Arthrobacter sp. SPG utilized 2-nitrobenzoate as its sole source of carbon and energy and degraded it with accumulation of stoichiometric amounts of nitrite ions. Salicylate and catechol were detected as metabolites of the 2-nitrobenzoate degradation using high performance liquid chromatography and gas chromatography-mass spectrometry. Enzyme activities for 2-nitrobenzoate-2-monooxygenase, salicylate hydroxylase, and catechol-1,2-dioxygenase were detected in the crude extracts of the 2-nitrobenzoate-induced cells of strain SPG. The 2-nitrobenzoate-monooxygenase activity resulted in formation of salicylate and nitrite from 2-nitrobenzoate whereas salicylate hydroxylase catalyzed the conversion of salicylate to catechol. The ring-cleaving enzyme, catechol-1,2-dioxygenase cleaved catechol to cis, cis-muconic acid. Cells of strain SPG were able to degrade 2-nitrobenzoate in sterile as well as non-sterile soil microcosms. The results of microcosm studies showed that strain SPG degraded more than 90% of 2-nitrobenzoate within 10-12 days. This study clearly shows that Arthrobacter sp. SPG degraded 2-nitrobenzoate via a new pathway with formation of salicylate and catechol as metabolites. Arthrobacter sp. SPG may be used for bioremediation of 2-nitrobenzoate-contaminated sites due to its ability to degrade 2-nitrobenzoate in soil.

  1. New metabolic pathway for degradation of 2-nitrobenzoate by Arthrobacter sp. SPG

    Science.gov (United States)

    Arora, Pankaj K.; Sharma, Ashutosh

    2015-01-01

    Arthrobacter sp. SPG utilized 2-nitrobenzoate as its sole source of carbon and energy and degraded it with accumulation of stoichiometric amounts of nitrite ions. Salicylate and catechol were detected as metabolites of the 2-nitrobenzoate degradation using high performance liquid chromatography and gas chromatography–mass spectrometry. Enzyme activities for 2-nitrobenzoate-2-monooxygenase, salicylate hydroxylase, and catechol-1,2-dioxygenase were detected in the crude extracts of the 2-nitrobenzoate-induced cells of strain SPG. The 2-nitrobenzoate-monooxygenase activity resulted in formation of salicylate and nitrite from 2-nitrobenzoate, whereas salicylate hydroxylase catalyzed the conversion of salicylate to catechol. The ring-cleaving enzyme, catechol-1,2-dioxygenase cleaved catechol to cis,cis-muconic acid. Cells of strain SPG were able to degrade 2-nitrobenzoate in sterile as well as non-sterile soil microcosms. The results of microcosm studies showed that strain SPG degraded more than 90% of 2-nitrobenzoate within 10–12 days. This study clearly shows that Arthrobacter sp. SPG degraded 2-nitrobenzoate via a new pathway with formation of salicylate and catechol as metabolites. Arthrobacter sp. SPG may be used for bioremediation of 2-nitrobenzoate-contaminated sites due to its ability to degrade 2-nitrobenzoate in soil. PMID:26082768

  2. Nicotine Dehydrogenase Complexed with 6-Hydroxypseudooxynicotine Oxidase Involved in the Hybrid Nicotine-Degrading Pathway in Agrobacterium tumefaciens S33.

    Science.gov (United States)

    Li, Huili; Xie, Kebo; Yu, Wenjun; Hu, Liejie; Huang, Haiyan; Xie, Huijun; Wang, Shuning

    2016-01-04

    Nicotine, a major toxic alkaloid in tobacco wastes, is degraded by bacteria, mainly via pyridine and pyrrolidine pathways. Previously, we discovered a new hybrid of the pyridine and pyrrolidine pathways in Agrobacterium tumefaciens S33 and characterized its key enzyme 6-hydroxy-3-succinoylpyridine (HSP) hydroxylase. Here, we purified the nicotine dehydrogenase initializing the nicotine degradation from the strain and found that it forms a complex with a novel 6-hydroxypseudooxynicotine oxidase. The purified complex is composed of three different subunits encoded by ndhAB and pno, where ndhA and ndhB overlap by 4 bp and are ∼26 kb away from pno. As predicted from the gene sequences and from chemical analyses, NdhA (82.4 kDa) and NdhB (17.1 kDa) harbor a molybdopterin cofactor and two [2Fe-2S] clusters, respectively, whereas Pno (73.3 kDa) harbors an flavin mononucleotide and a [4Fe-4S] cluster. Mutants with disrupted ndhA or ndhB genes did not grow on nicotine but grew well on 6-hydroxynicotine and HSP, whereas the pno mutant did not grow on nicotine or 6-hydroxynicotine but grew well on HSP, indicating that NdhA and NdhB are responsible for initialization of nicotine oxidation. We successfully expressed pno in Escherichia coli and found that the recombinant Pno presented 2,6-dichlorophenolindophenol reduction activity when it was coupled with 6-hydroxynicotine oxidation. The determination of reaction products catalyzed by the purified enzymes or mutants indicated that NdhAB catalyzed nicotine oxidation to 6-hydroxynicotine, whereas Pno oxidized 6-hydroxypseudooxynicotine to 6-hydroxy-3-succinoylsemialdehyde pyridine. These results provide new insights into this novel hybrid pathway of nicotine degradation in A. tumefaciens S33.

  3. Novel degradation pathway and kinetic analysis for buprofezin removal by newly isolated Bacillus sp.

    Science.gov (United States)

    Wang, Guangli; Xu, Dayong; Xiong, Minghua; Zhang, Hui; Li, Feng; Liu, Yuan

    2016-09-15

    Given the intensive and widespread application of the pesticide, buprofezin, its environmental residues potentially pose a problem; yet little is known about buprofezin's kinetic and metabolic behaviors. In this study, a novel gram-positive strain, designated BF-5, isolated from aerobic activated sludge, was found to be capable of metabolizing buprofezin as its sole energy, carbon, and nitrogen source. Based on its physiological and biochemical characteristics, other aspects of its phenotype, and a phylogenetic analysis, strain BF-5 was identified as Bacillus sp. This study investigated the effect of culture conditions on bacterial growth and substrate degradation, such as pH, temperature, initial concentration, different nitrogen source, and additional nitrogen sources as co-substrates. The degradation rate parameters, qmax, Ks, Ki and Sm were determined to be 0.6918 h(-1), 105.4 mg L(-1), 210.5 mg L(-1), and 148.95 mg L(-1) respectively. The capture of unpublished potential metabolites by gas chromatography-mass spectrometry (GC-MS) analysis has led to the proposal of a novel degradation pathway. Taken together, our results clarify buprofezin's biodegradation pathway(s) and highlight the promising potential of strain BF-5 in bioremediation of buprofezin-contaminated environments. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Characterization of stress degradation products of duloxetine hydrochloride employing LC-UV/PDA and LC-MS/TOF studies.

    Science.gov (United States)

    Chadha, Renu; Bali, Alka; Bansal, Gulshan

    2016-03-20

    Duloxetine HCl was subjected to forced degradation under conditions of hydrolysis (neutral, acidic and alkaline), oxidation, photolysis and thermal stress, as suggested in the ICH guideline Q1A(R2). The drug showed significant degradation under acidic, alkaline and aqueous hydrolytic as well as photolytic conditions. The drug remained stable under thermal and oxidative stress conditions. In total, seventeen degradation products (I-XVII) were formed under varied conditions, which could be separated by chromatography of respective degraded solutions on C18 (250 mm×4.6 mm; 5 μ, Nulceodur) column using isocratic elution method. Detection wavelength was selected as 290 nm. MS/TOF accurate mass studies were carried out to establish the complete fragmentation pathway of the drug and degradation products, which, in turn, was utilized in characterization of the products. The degradation pathway of the drug leading to generation of fifteen products I-X, XII-XIII, XV-XVII was postulated and this has not been reported so far.

  5. From ether to acid: A plausible degradation pathway of glycerol dialkyl glycerol tetraethers

    Science.gov (United States)

    Liu, Xiao-Lei; Birgel, Daniel; Elling, Felix J.; Sutton, Paul A.; Lipp, Julius S.; Zhu, Rong; Zhang, Chuanlun; Könneke, Martin; Peckmann, Jörn; Rowland, Steven J.; Summons, Roger E.; Hinrichs, Kai-Uwe

    2016-06-01

    Glycerol dialkyl glycerol tetraethers (GDGTs) are ubiquitous microbial lipids with extensive demonstrated and potential roles as paleoenvironmental proxies. Despite the great attention they receive, comparatively little is known regarding their diagenetic fate. Putative degradation products of GDGTs, identified as hydroxyl and carboxyl derivatives, were detected in lipid extracts of marine sediment, seep carbonate, hot spring sediment and cells of the marine thaumarchaeon Nitrosopumilus maritimus. The distribution of GDGT degradation products in environmental samples suggests that both biotic and abiotic processes act as sinks for GDGTs. More than a hundred newly recognized degradation products afford a view of the stepwise degradation of GDGT via (1) ether bond hydrolysis yielding hydroxyl isoprenoids, namely, GDGTol (glycerol dialkyl glycerol triether alcohol), GMGD (glycerol monobiphytanyl glycerol diether), GDD (glycerol dibiphytanol diether), GMM (glycerol monobiphytanol monoether) and bpdiol (biphytanic diol); (2) oxidation of isoprenoidal alcohols into corresponding carboxyl derivatives and (3) chain shortening to yield C39 and smaller isoprenoids. This plausible GDGT degradation pathway from glycerol ethers to isoprenoidal fatty acids provides the link to commonly detected head-to-head linked long chain isoprenoidal hydrocarbons in petroleum and sediment samples. The problematic C80 to C82 tetraacids that cause naphthenate deposits in some oil production facilities can be generated from H-shaped glycerol monoalkyl glycerol tetraethers (GMGTs) following the same process, as indicated by the distribution of related derivatives in hydrothermally influenced sediments.

  6. Oxidative degradation of N-Nitrosopyrrolidine by the ozone/UV process: Kinetics and pathways.

    Science.gov (United States)

    Chen, Zhi; Fang, Jingyun; Fan, Chihhao; Shang, Chii

    2016-05-01

    N-Nitrosopyrrolidine (NPYR) is an emerging contaminant in drinking water and wastewater. The degradation kinetics and mechanisms of NPYR degradation by the O3/UV process were investigated and compared with those of UV direct photolysis and ozonation. A synergistic effect of ozone and UV was observed in the degradation of NPYR due to the accelerated production of OH• by ozone photolysis. This effect was more pronounced at higher ozone dosages. The second-order rate constants of NPYR reacting with OH• and ozone was determined to be 1.38 (± 0.05) × 10(9) M(-1) s(-1) and 0.31 (± 0.02) M(-1) s(-1), respectively. The quantum yield by direct UV photolysis was 0.3 (± 0.01). An empirical model using Rct (the ratio of the exposure of OH• to that of ozone) was established for NPYR degradation in treated drinking water and showed that the contributions of direct UV photolysis and OH• oxidation on NPYR degradation were both significant. As the reaction proceeded, the contribution by OH• became less important due to the exhausting of ozone. Nitrate was the major product in the O3/UV process by two possible pathways. One is through the cleavage of nitroso group to form NO• followed by hydrolysis, and the other is the oxidation of the intermediates of amines by ozonation.

  7. Force degradation behavior of glucocorticoid deflazacort by UPLC: isolation, identification and characterization of degradant by FTIR, NMR and mass analysis

    Science.gov (United States)

    Deshmukh, Rajesh; Sharma, Lata; Tekade, Muktika; Kesharwani, Prashant; Trivedi, Piyush; Tekade, Rakesh K.

    2016-01-01

    Abstract In this investigation, sensitive and reproducible methods are described for quantitative determination of deflazacort in the presence of its degradation product. The method was based on high performance liquid chromatography of the drug from its degradation product on reverse phase using Acquity UPLC BEH C18 columns (1.7 µm, 2.1 mm × 150 mm) using acetonitrile and water (40:60 V/V) at a flow rate of 0.2 mL/minute in UPLC. UV detection was performed at 240.1 nm. Deflazacort was subjected to oxidative, acid, base, hydrolytic, thermal and photolytic degradation. The drug was found to be stable in water and thermal stress, as well as under neutral stress conditions. However, forced-degradation study performed on deflazacort showed that the drug degraded under alkaline, acid and photolytic stress. The degradation products were well resolved from the main peak, which proved the stability-indicating power of the method. The developed method was validated as per ICH guidelines with respect to accuracy, linearity, limit of detection, limit of quantification, accuracy, precision and robustness, selectivity and specificity. Apart from the aforementioned, the results of the present study also emphasize the importance of isolation characterization and identification of degradant. Hence, an attempt was made to identify the degradants in deflazacort. One of the degradation products of deflazacort was isolated and identified by the FTIR, NMR and LC-MS study. PMID:28276670

  8. PINK1-Parkin pathway activity is regulated by degradation of PINK1 in the mitochondrial matrix.

    Directory of Open Access Journals (Sweden)

    Ruth E Thomas

    Full Text Available Loss-of-function mutations in PINK1, which encodes a mitochondrially targeted serine/threonine kinase, result in an early-onset heritable form of Parkinson's disease. Previous work has shown that PINK1 is constitutively degraded in healthy cells, but selectively accumulates on the surface of depolarized mitochondria, thereby initiating their autophagic degradation. Although PINK1 is known to be a cleavage target of several mitochondrial proteases, whether these proteases account for the constitutive degradation of PINK1 in healthy mitochondria remains unclear. To explore the mechanism by which PINK1 is degraded, we performed a screen for mitochondrial proteases that influence PINK1 abundance in the fruit fly Drosophila melanogaster. We found that genetic perturbations targeting the matrix-localized protease Lon caused dramatic accumulation of processed PINK1 species in several mitochondrial compartments, including the matrix. Knockdown of Lon did not decrease mitochondrial membrane potential or trigger activation of the mitochondrial unfolded protein stress response (UPRmt, indicating that PINK1 accumulation in Lon-deficient animals is not a secondary consequence of mitochondrial depolarization or the UPRmt. Moreover, the influence of Lon on PINK1 abundance was highly specific, as Lon inactivation had little or no effect on the abundance of other mitochondrial proteins. Further studies indicated that the processed forms of PINK1 that accumulate upon Lon inactivation are capable of activating the PINK1-Parkin pathway in vivo. Our findings thus suggest that Lon plays an essential role in regulating the PINK1-Parkin pathway by promoting the degradation of PINK1 in the matrix of healthy mitochondria.

  9. Interleukin-1 Acts via the JNK-2 Signaling Pathway to Induce Aggrecan Degradation by Human Chondrocytes.

    Science.gov (United States)

    Ismail, Heba M; Yamamoto, Kazuhiro; Vincent, Tonia L; Nagase, Hideaki; Troeberg, Linda; Saklatvala, Jeremy

    2015-07-01

    Aggrecan enables articular cartilage to bear load and resist compression. Aggrecan loss occurs early in osteoarthritis and rheumatoid arthritis and can be induced by inflammatory cytokines such as interleukin-1 (IL-1). IL-1 induces cleavage of specific aggrecans characteristic of the ADAMTS proteinases. The aim of this study was to identify the intracellular signaling pathways by which IL-1 causes aggrecan degradation by human chondrocytes and to investigate how aggrecanase activity is controlled by chondrocytes. We developed a cell-based assay combining small interfering RNA (siRNA)-induced knockdown with aggrecan degradation assays. Human articular chondrocytes were overlaid with bovine aggrecan after transfection with siRNAs against molecules of the IL-1 signaling pathway. After IL-1 stimulation, released aggrecan fragments were detected with AGEG and ARGS neoepitope antibodies. Aggrecanase activity and tissue inhibitor of metalloproteinases 3 levels were measured by enzyme-linked immunosorbent assay. Low-density lipoprotein receptor-related protein 1 (LRP-1) shedding was analyzed by Western blotting. ADAMTS-5 is a major aggrecanase in human chondrocytes, regulating aggrecan degradation in response to IL-1. The tumor necrosis factor receptor-associated 6 (TRAF-6)/transforming growth factor β-activated kinase 1 (TAK-1)/MKK-4 signaling axis is essential for IL-1-induced aggrecan degradation, while NF-κB is not. Of the 3 MAPKs (ERK, p38, and JNK), only JNK-2 showed a significant role in aggrecan degradation. Chondrocytes constitutively secreted aggrecanase, which was continuously endocytosed by LRP-1, keeping the extracellular level of aggrecanase low. IL-1 induced aggrecanase activity in the medium in a JNK-2-dependent manner, possibly by reducing aggrecanase endocytosis, because IL-1 caused JNK-2-dependent shedding of LRP-1. The signaling axis TRAF-6/TAK-1/MKK-4/JNK-2 mediates IL-1-induced aggrecanolysis. The level of aggrecanase is controlled by its

  10. Isolation and characterization of diesel oil degrading indigenous ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... achieved 86.94% of diesel degradation in 2 weeks. Additional degradation ... spills, especially in soil contamination have prompted research on ... hydrocarbons are natural products, it is not surprising to find organisms that ...

  11. Involvement of two latex-clearing proteins during rubber degradation and insights into the subsequent degradation pathway revealed by the genome sequence of Gordonia polyisoprenivorans strain VH2.

    Science.gov (United States)

    Hiessl, Sebastian; Schuldes, Jörg; Thürmer, Andrea; Halbsguth, Tobias; Bröker, Daniel; Angelov, Angel; Liebl, Wolfgang; Daniel, Rolf; Steinbüchel, Alexander

    2012-04-01

    The increasing production of synthetic and natural poly(cis-1,4-isoprene) rubber leads to huge challenges in waste management. Only a few bacteria are known to degrade rubber, and little is known about the mechanism of microbial rubber degradation. The genome of Gordonia polyisoprenivorans strain VH2, which is one of the most effective rubber-degrading bacteria, was sequenced and annotated to elucidate the degradation pathway and other features of this actinomycete. The genome consists of a circular chromosome of 5,669,805 bp and a circular plasmid of 174,494 bp with average GC contents of 67.0% and 65.7%, respectively. It contains 5,110 putative protein-coding sequences, including many candidate genes responsible for rubber degradation and other biotechnically relevant pathways. Furthermore, we detected two homologues of a latex-clearing protein, which is supposed to be a key enzyme in rubber degradation. The deletion of these two genes for the first time revealed clear evidence that latex-clearing protein is essential for the microbial utilization of rubber. Based on the genome sequence, we predict a pathway for the microbial degradation of rubber which is supported by previous and current data on transposon mutagenesis, deletion mutants, applied comparative genomics, and literature search.

  12. KCTD1 suppresses canonical Wnt signaling pathway by enhancing β-catenin degradation.

    Directory of Open Access Journals (Sweden)

    Xinxin Li

    Full Text Available The canonical Wnt signaling pathway controls normal embryonic development, cellular proliferation and growth, and its aberrant activity results in human carcinogenesis. The core component in regulation of this pathway is β-catenin, but molecular regulation mechanisms of β-catenin stability are not completely known. Here, our recent studies have shown that KCTD1 strongly inhibits TCF/LEF reporter activity. Moreover, KCTD1 interacted with β-catenin both in vivo by co-immunoprecipitation as well as in vitro through GST pull-down assays. We further mapped the interaction regions to the 1-9 armadillo repeats of β-catenin and the BTB domain of KCTD1, especially Position Ala-30 and His-33. Immunofluorescence analysis indicated that KCTD1 promotes the cytoplasmic accumulation of β-catenin. Furthermore, protein stability assays revealed that KCTD1 enhances the ubiquitination/degradation of β-catenin in a concentration-dependent manner in HeLa cells. And the degradation of β-catenin mediated by KCTD1 was alleviated by the proteasome inhibitor, MG132. In addition, KCTD1-mediated β-catenin degradation was dependent on casein kinase 1 (CK1- and glycogen synthase kinase-3β (GSK-3β-mediated phosphorylation and enhanced by the E3 ubiquitin ligase β-transducin repeat-containing protein (β-TrCP. Moreover, KCTD1 suppressed the expression of endogenous Wnt downstream genes and transcription factor AP-2α. Finally, we found that Wnt pathway member APC and tumor suppressor p53 influence KCTD1-mediated downregulation of β-catenin. These results suggest that KCTD1 functions as a novel inhibitor of Wnt signaling pathway.

  13. Characterization of degraded soils in the humid Ethiopian highlands

    NARCIS (Netherlands)

    Tebebu, Tigist Y.; Bayabil, Haimanote K.; Stoof, C.R.; Giri, Shree K.; Gessess, A.A.; Tilahun, Seifu A.; Steenhuis, Tammo S.

    2016-01-01

    Hard pan is a major cause of land degradation that affects agricultural productivity in developing countries. However, relatively little is known about the interaction of land degradation and hardpans. The objective of this study was, therefore, to investigate soil degradation and the formation of h

  14. Arginine deiminase pathway genes and arginine degradation variability in Oenococcus oeni strains.

    Science.gov (United States)

    Araque, Isabel; Gil, Joana; Carreté, Ramon; Constantí, Magda; Bordons, Albert; Reguant, Cristina

    2016-03-01

    Trace amounts of the carcinogenic ethyl carbamate can appear in wine as a result of a reaction between ethanol and citrulline, which is produced from arginine degradation by some bacteria used in winemaking. In this study, arginine deiminase (ADI) pathway genes were evaluated in 44 Oenococcus oeni strains from wines originating from several locations in order to establish the relationship between the ability of a strain to degrade arginine and the presence of related genes. To detect the presence of arc genes of the ADI pathway in O. oeni, pairs of primers were designed to amplify arcA, arcB, arcC and arcD1 sequences. All strains contained these four genes. The same primers were used to confirm the organization of these genes in an arcABCD1 operon. Nevertheless, considerable variability in the ability to degrade arginine among these O. oeni strains was observed. Therefore, despite the presence of the arc genes in all strains, the expression patterns of individual genes must be strain dependent and influenced by the different wine conditions. Additionally, the presence of arc genes was also determined in the 57 sequenced strains of O. oeni available in GenBank, and the complete operon was found in 83% of strains derived from wine. The other strains were found to lack the arcB, arcC and arcD genes, but all contained sequences homologous to arcA, and some of them had also ADI activity.

  15. Degradation pathways of low-ethoxylated nonylphenols by isolated bacteria using an improved method.

    Science.gov (United States)

    Zhang, Yu; Gu, Xin; Zhang, Jing; Yang, Min

    2014-01-01

    Nonylphenol ethoxylates (NPEOs) with low ethoxylation degree (NPav₂EO; containing two ethoxy units on average) and estrogenic properties are the intermediate products of nonionic surfactant NPEOs. To better understand the environmental fate of low-ethoxylated NPEOs, phylogenetically diverse low-ethoxylated NPEO-degrading bacteria were isolated from activated sludge using gellan gum as the gelling reagent. Four isolates belonging to four genera, i.e., Pseudomonas sp. NP522b in γ-Proteobacteria, Variovorax sp. NP427b and Ralstonia sp. NP47a in β-Proteobacteria, and Sphingomonas sp. NP42a in α-Proteobacteria were acquired. Ralstonia sp. NP47a or Sphingomonas sp. NP42a, have not been reported for the degradation of low-ethoxylated NPEOs previously. The biotransformation pathways of these isolates were investigated. The first three strains (NP522b, NP427b, and NP47a) exhibited high NPav₂EO oxidation ability by oxidizing the polyethoxy (EO) chain to form low-ethoxylated nonylphenoxy carboxylates, and then further oxidizing the alkyl chain to form carboxyalkylphenol polyethoxycarboxylates. Furthermore, Sphingomonas sp. NP42a degraded NPav2EO through a nonoxidative pathway with nonylphenol monoethoxylate as the dominant product.

  16. Genetic immunization based on the ubiquitin-fusion degradation pathway against Trypanosoma cruzi

    Energy Technology Data Exchange (ETDEWEB)

    Chou, Bin [Department of Microbiology and Immunology, Faculty of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-0180 (Japan); Department of Parasitology, Graduate School of Medical Science, Kyushu University, Fukuoka 812-8582 (Japan); Hiromatsu, Kenji, E-mail: khiromatsu@fukuoka-u.ac.jp [Department of Microbiology and Immunology, Faculty of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-0180 (Japan); Hisaeda, Hajime; Duan, Xuefeng; Imai, Takashi [Department of Parasitology, Graduate School of Medical Science, Kyushu University, Fukuoka 812-8582 (Japan); Murata, Shigeo; Tanaka, Keiji [Department of Molecular Oncology, The Tokyo Metropolitan Institute of Medical Science, Tokyo 113-8613 (Japan); Himeno, Kunisuke [Department of Parasitology, Graduate School of Medical Science, Kyushu University, Fukuoka 812-8582 (Japan)

    2010-02-12

    Cytotoxic CD8{sup +} T cells are particularly important to the development of protective immunity against the intracellular protozoan parasite, Trypanosoma cruzi, the etiological agent of Chagas disease. We have developed a new effective strategy of genetic immunization by activating CD8{sup +} T cells through the ubiquitin-fusion degradation (UFD) pathway. We constructed expression plasmids encoding the amastigote surface protein-2 (ASP-2) of T. cruzi. To induce the UFD pathway, a chimeric gene encoding ubiquitin fused to ASP-2 (pUB-ASP-2) was constructed. Mice immunized with pUB-ASP-2 presented lower parasitemia and longer survival period, compared with mice immunized with pASP-2 alone. Depletion of CD8{sup +} T cells abolished protection against T. cruzi in mice immunized with pUB-ASP-2 while depletion of CD4{sup +} T cells did not influence the effective immunity. Mice deficient in LMP2 or LMP7, subunits of immunoproteasomes, were not able to develop protective immunity induced. These results suggest that ubiquitin-fused antigens expressed in antigen-presenting cells were effectively degraded via the UFD pathway, and subsequently activated CD8{sup +} T cells. Consequently, immunization with pUB-ASP-2 was able to induce potent protective immunity against infection of T. cruzi.

  17. Tyrosol degradation via the homogentisic acid pathway in a newly isolated Halomonas strain from olive processing effluents

    OpenAIRE

    Liebgott, Pierre-Pol; Labat, Marc; Amouric, Agnès; Tholozan, Jean-Luc; Lorquin, Jean

    2008-01-01

    To isolate a new Halomonas sp. strain capable of degrading tyrosol, a toxic compound present in olive mill wastewater, through the homogentisic acid (HGA) pathway. A moderately halophilic Gram-negative bacterium belonging to the Halomonas genus and designated strain TYRC17 was isolated from olive processing effluents. This strain was able to completely degrade tyrosol (2-(p-hydroxyphenyl)-ethanol), a toxic compound found in such effluent. Tyrosol degradation begins by an oxidation to 4-hydrox...

  18. Molecular characterization of a toluene-degrading methanogenic consortium.

    Science.gov (United States)

    Ficker, M; Krastel, K; Orlicky, S; Edwards, E

    1999-12-01

    A toluene-degrading methanogenic consortium enriched from creosote-contaminated aquifer material was maintained on toluene as the sole carbon and energy source for 10 years. The species in the consortium were characterized by using a molecular approach. Total genomic DNA was isolated, and 16S rRNA genes were amplified by using PCR performed with kingdom-specific primers that were specific for 16S rRNA genes from either members of the kingdom Bacteria or members of the kingdom Archaea. A total of 90 eubacterial clones and 75 archaeal clones were grouped by performing a restriction fragment length polymorphism (RFLP) analysis. Six eubacterial sequences and two archaeal sequences were found in the greatest abundance (in six or more clones) based on the RFLP analysis. The relative abundance of each putative species was estimated by using fluorescent in situ hybridization (FISH), and the presence of putative species was determined qualitatively by performing slot blot hybridization with consortium DNA. Both archaeal species and two of the six eubacterial species were detected in the DNA and FISH hybridization experiments. A phylogenetic analysis of these four dominant organisms suggested that the two archaeal species are related to the genera Methanosaeta and Methanospirillum. One of the eubacterial species is related to the genus Desulfotomaculum, while the other is not related to any previously described genus. By elimination, we propose that the last organism probably initiates the attack on toluene.

  19. Insulin-degrading enzyme is exported via an unconventional protein secretion pathway

    Directory of Open Access Journals (Sweden)

    Leissring Malcolm A

    2009-01-01

    Full Text Available Abstract Insulin-degrading enzyme (IDE is a ubiquitously expressed zinc-metalloprotease that degrades several pathophysiologically significant extracellular substrates, including insulin and the amyloid β-protein (Aβ, and accumulating evidence suggests that IDE dysfunction may be operative in both type 2 diabetes mellitus and Alzheimer disease (AD. Although IDE is well known to be secreted by a variety of cell types, the underlying trafficking pathway(s remain poorly understood. To address this topic, we investigated the effects of known inhibitors or stimulators of protein secretion on the secretion of IDE from murine hepatocytes and HeLa cells. IDE secretion was found to be unaffected by the classical secretion inhibitors brefeldin A (BFA, monensin, or nocodazole, treatments that readily inhibited the secretion of α1-antitrypsin (AAT overexpressed in the same cells. Using a novel cell-based Aβ-degradation assay, we show further that IDE secretion was similarly unaffected by multiple stimulators of protein secretion, including glyburide and 3'-O-(4-benzoylbenzoyl-ATP (Bz-ATP. The calcium ionophore, A23187, increased extracellular IDE activity, but only under conditions that also elicited cytotoxicity. Our results provide the first biochemical evidence that IDE export is not dependent upon the classical secretion pathway, thereby identifying IDE as a novel member of the select class of unconventionally secreted proteins. Further elucidation of the mechanisms underlying IDE secretion, which would be facilitated by the assays described herein, promises to uncover processes that might be defective in disease or manipulated for therapeutic benefit.

  20. Ubiquitin initiates sorting of Golgi and plasma membrane proteins into the vacuolar degradation pathway

    Directory of Open Access Journals (Sweden)

    Scheuring David

    2012-09-01

    Full Text Available Abstract Background In yeast and mammals, many plasma membrane (PM proteins destined for degradation are tagged with ubiquitin. These ubiquitinated proteins are internalized into clathrin-coated vesicles and are transported to early endosomal compartments. There, ubiquitinated proteins are sorted by the endosomal sorting complex required for transport (ESCRT machinery into the intraluminal vesicles of multivesicular endosomes. Degradation of these proteins occurs after endosomes fuse with lysosomes/lytic vacuoles to release their content into the lumen. In plants, some PM proteins, which cycle between the PM and endosomal compartments, have been found to be ubiquitinated, but it is unclear whether ubiquitin is sufficient to mediate internalization and thus acts as a primary sorting signal for the endocytic pathway. To test whether plants use ubiquitin as a signal for the degradation of membrane proteins, we have translationally fused ubiquitin to different fluorescent reporters for the plasma membrane and analyzed their transport. Results Ubiquitin-tagged PM reporters localized to endosomes and to the lumen of the lytic vacuole in tobacco mesophyll protoplasts and in tobacco epidermal cells. The internalization of these reporters was significantly reduced if clathrin-mediated endocytosis was inhibited by the coexpression of a mutant of the clathrin heavy chain, the clathrin hub. Surprisingly, a ubiquitin-tagged reporter for the Golgi was also transported into the lumen of the vacuole. Vacuolar delivery of the reporters was abolished upon inhibition of the ESCRT machinery, indicating that the vacuolar delivery of these reporters occurs via the endocytic transport route. Conclusions Ubiquitin acts as a sorting signal at different compartments in the endomembrane system to target membrane proteins into the vacuolar degradation pathway: If displayed at the PM, ubiquitin triggers internalization of PM reporters into the endocytic transport route

  1. Electrochemical treatment of trypan blue synthetic wastewater and its degradation pathway

    Directory of Open Access Journals (Sweden)

    ANANTHA N. SUBBA RAO

    2013-11-01

    Full Text Available The trypan blue (TB dye synthetic wastewater was treated in presence of chloride ions by electrochemical method. The effect of current density, pH, initial concentration of dye and supporting electrolyte on color and COD removal were investigated. The UV-Vis ab­sorption intensity, chemical oxygen demand (COD, cyclic voltammetry (CV, Fourier transform- infrared spectroscopy (FT-IR, gas chromatography – mass spectrometry (GC-MS analysis were conducted to investigate the kinetics and degradation pathway of TB dye.

  2. The Branched-Chain Dodecylbenzene Sulfonate Degradation Pathway of Pseudomonas aeruginosa W51D Involves a Novel Route for Degradation of the Surfactant Lateral Alkyl Chain

    OpenAIRE

    Campos-García, Jesús; Esteve, Abraham; Vázquez-Duhalt, Rafael; Ramos, Juán Luis; Soberón-Chávez, Gloria

    1999-01-01

    Pseudomonas aeruginosa W51D is able to grow by using branched-chain dodecylbenzene sulfonates (B-DBS) or the terpenic alcohol citronellol as a sole source of carbon. A mutant derived from this strain (W51M1) is unable to degrade citronellol but still grows on B-DBS, showing that the citronellol degradation route is not the main pathway involved in the degradation of the surfactant alkyl moiety. The structures of the main B-DBS isomers and of some intermediates were identified by gas chromatog...

  3. The Branched-Chain Dodecylbenzene Sulfonate Degradation Pathway of Pseudomonas aeruginosa W51D Involves a Novel Route for Degradation of the Surfactant Lateral Alkyl Chain

    Science.gov (United States)

    Campos-García, Jesús; Esteve, Abraham; Vázquez-Duhalt, Rafael; Ramos, Juán Luis; Soberón-Chávez, Gloria

    1999-01-01

    Pseudomonas aeruginosa W51D is able to grow by using branched-chain dodecylbenzene sulfonates (B-DBS) or the terpenic alcohol citronellol as a sole source of carbon. A mutant derived from this strain (W51M1) is unable to degrade citronellol but still grows on B-DBS, showing that the citronellol degradation route is not the main pathway involved in the degradation of the surfactant alkyl moiety. The structures of the main B-DBS isomers and of some intermediates were identified by gas chromatography-mass spectrometric analysis, and a possible catabolic route is proposed. PMID:10427075

  4. Characterization of cyanobacterial hydrocarbon composition and distribution of biosynthetic pathways.

    Directory of Open Access Journals (Sweden)

    R Cameron Coates

    Full Text Available Cyanobacteria possess the unique capacity to naturally produce hydrocarbons from fatty acids. Hydrocarbon compositions of thirty-two strains of cyanobacteria were characterized to reveal novel structural features and insights into hydrocarbon biosynthesis in cyanobacteria. This investigation revealed new double bond (2- and 3-heptadecene and methyl group positions (3-, 4- and 5-methylheptadecane for a variety of strains. Additionally, results from this study and literature reports indicate that hydrocarbon production is a universal phenomenon in cyanobacteria. All cyanobacteria possess the capacity to produce hydrocarbons from fatty acids yet not all accomplish this through the same metabolic pathway. One pathway comprises a two-step conversion of fatty acids first to fatty aldehydes and then alkanes that involves a fatty acyl ACP reductase (FAAR and aldehyde deformylating oxygenase (ADO. The second involves a polyketide synthase (PKS pathway that first elongates the acyl chain followed by decarboxylation to produce a terminal alkene (olefin synthase, OLS. Sixty-one strains possessing the FAAR/ADO pathway and twelve strains possessing the OLS pathway were newly identified through bioinformatic analyses. Strains possessing the OLS pathway formed a cohesive phylogenetic clade with the exception of three Moorea strains and Leptolyngbya sp. PCC 6406 which may have acquired the OLS pathway via horizontal gene transfer. Hydrocarbon pathways were identified in one-hundred-forty-two strains of cyanobacteria over a broad phylogenetic range and there were no instances where both the FAAR/ADO and the OLS pathways were found together in the same genome, suggesting an unknown selective pressure maintains one or the other pathway, but not both.

  5. Elimination of paternal mitochondria through the lysosomal degradation pathway in C.elegans

    Institute of Scientific and Technical Information of China (English)

    Qinghua Zhou; Haimin Li; Ding Xue

    2011-01-01

    In mammals,the inheritance of mitochondrion and its DNA (mtDNA) is strictly maternal,despite the fact that a sperm can inject up to 100 functional mitochondria into the oocyte during fertilization.The mechanisms responsible for the elimination of the paternal mitochondria remain largely unknown.We report here that this paternal mitochondrial elimination process is conserved in Caenorhabditis elegans,and that the lysosomal pathway actively participates in this process.Molecular and cell biological analyses indicate that in wild-type animals paternal mitoehondria and mtDNA are destroyed within two hours after fertilization.In animals with compromised lysosomes,paternal mitochondria persist until late embryonic stages.Therefore,the lysosomal pathway plays an important role in degrading paternal mitochondria introduced into the oocyte during fertilization.Our study indicates that C.elegans is an excellent animal model for understanding and dissecting this conserved biological process critical for animal development and reproduction.

  6. Study of the forced degradation behavior of prasugrel hydrochloride by liquid chromatography with mass spectrometry and liquid chromatography with NMR detection and prediction of the toxicity of the characterized degradation products.

    Science.gov (United States)

    Singh, Dilip Kumar; Sahu, Archana; Handa, Tarun; Narayanam, Mallikarjun; Singh, Saranjit

    2015-09-01

    Prasugrel was subjected to forced degradation studies under conditions of hydrolysis (acid, base, and neutral), photolysis, oxidation, and thermal stress. The drug showed liability in hydrolytic as well as oxidative conditions, resulting in a total of four degradation products. In order to characterize the latter, initially mass fragmentation pathway of the drug was established with the help of mass spectrometry/time-of-flight, multiple stage mass spectrometry and hydrogen/deuterium exchange data. The degradation products were then separated on a C18 column using a stability-indicating volatile buffer method, which was later extended to liquid chromatography-mass spectrometry studies. The latter highlighted that three degradation products had the same molecular mass, while one was different. To characterize all, their mass fragmentation pathways were established in the same manner as the drug. Subsequently, liquid chromatography-nuclear magnetic resonance (NMR) spectroscopy data were collected. Proton and correlation liquid chromatography with NMR spectroscopy studies highlighted existence of diastereomeric behavior in one pair of degradation products. Lastly, toxicity prediction by computer-assisted technology (TOPKAT) and deductive estimation of risk from existing knowledge (DEREK) software were employed to assess in silico toxicity of the characterized degradation products.

  7. Def defines a conserved nucleolar pathway that leads p53 to proteasome-independent degradation

    Institute of Scientific and Technical Information of China (English)

    Ting Tao; Hui Shi; Yihong Guan; Delai Huang; Ye Chen; David P Lane; Jun Chen

    2013-01-01

    p53 protein turnover through the ubiquitination pathway is a vital mechanism in the regulation of its transcriptional activity; however,little is known about p53 turnover through proteasome-independent pathway(s).The digestive organ expansion factor (Def) protein is essential for the development of digestive organs.In zebrafish,loss of function of defselectively upregulates the expression of p53 response genes,which raises a question as to what is the relationship between Def and p53.We report here that Def is a nucleolar protein and that loss of function of defleads to the upregulation of p53 protein,which surprisingly accumulates in the nucleoli.Our extensive studies have demonstrated that Def can mediate the degradation of p53 protein and that this process is independent of the proteasome pathway,but dependent on the activity of Calpain3,a cysteine protease.Our findings define a novel nucleolar pathway that regulates the turnover function of p53,which will advance our understanding of p53's role in organogenesis and tumorigenesis.

  8. Microbial Degradation of Polycyclic Aromatic Hydrocarbons and Characterization of Bacteria

    Science.gov (United States)

    Tikilili, P. V.; Chirwa, E. M. N.

    2010-01-01

    Biodegradation of polycyclic aromatic hydrocarbons was studied. Naphthalene was used as a model compound to represent these compounds. Low initial concentrations of naphthalene in a range of 30-60 mg/L were completely degraded after incubation for 15 hrs by consortia from a landfill soil while consortia from minewater took more that 29 hrs to reach complete degradation.

  9. Kinetics and pathways of ibuprofen degradation by the UV/chlorine advanced oxidation process.

    Science.gov (United States)

    Xiang, Yingying; Fang, Jingyun; Shang, Chii

    2016-03-01

    The UV/chlorine advanced oxidation process (AOP), which forms reactive species such as hydroxyl radicals (HO) and reactive chlorine species (RCS) such as chlorine atoms (Cl) and Cl2(-), is being considered as an alternative to the UV/H2O2 AOP for the degradation of emerging contaminants. This study investigated the kinetics and pathways of the degradation of a recalcitrant pharmaceutical and personal care product (PPCP)-ibuprofen (IBP)-by the UV/chlorine AOP. The degradation of IBP followed the pseudo first-order kinetics. The first-order rate constant was 3.3 times higher in the UV/chlorine AOP than in the UV/H2O2 AOP for a given chemical molar dosage at pH 6. The first-order rate constant decreased from 3.1 × 10(-3) s(-1) to 5.5 × 10(-4) s(-1) with increasing pH from 6 to 9. Both HO and RCS contributed to the degradation, and the contribution of RCS increased from 22% to 30% with increasing pH from 6 to 9. The degradation was initiated by HO-induced hydroxylation and Cl-induced chlorine substitution, and sustained through decarboxylation, demethylation, chlorination and ring cleavage to form more stable products. Significant amounts of chlorinated intermediates/byproducts were formed from the UV/chlorine AOP, and four chlorinated products were newly identified. The yield of total organic chlorine (TOCl) was 31.6 μM after 90% degradation of 50 μM IBP under the experimental conditions. The known disinfection by-products (DBPs) comprised 17.4% of the TOCl. The effects of water matrix in filtered drinking water on the degradation were not significant, demonstrating the practicality of the UV/chlorine AOP for the control of some refractory PPCPs. However, the toxicity of the chlorinated products should be further assessed.

  10. Kinetics and reaction pathways of formaldehyde degradation using the UV-fenton method.

    Science.gov (United States)

    Liu, Xiangxuan; Liang, Jiantao; Wang, Xuanjun

    2011-05-01

    This study was based on the purpose of investigating the reaction rules of formaldehyde (HCHO) as an intermediate product in the degradation of many other organic wastewaters. The process conditions of UV-Fenton method for the degradation of the low concentrations of HCHO were studied in a batch photochemical reactor. The results showed that, when the original HCHO concentration was 30 mg/L, at an operating temperature of 23 degrees C, pH = 3, an H202 dosage of 68 mg/L, and an H2O2-to-Fe2+ mole ratio (H2O2:Fe2+) of 5, 91.89% of the HCHO was removed after 30 minutes. The degradation of HCHO in the UV-Fenton system was basically in accordance with the exponential decay. The kinetic study results showed that the reaction orders of HCHO, Fe2+, and H2O2 in the system were 1.054, 0.510, and 0.728, respectively, and the activation energy (Ea) was 9.85 kJ/mol. The comparison of UV/H2O2, Fenton, and UV-Fenton systems for the degradation of HCHO, and the results of iron catalyst tests showed that the mechanism of UV-Fenton on the degradation of HCHO was through a synergistic effect of Fe2+ and UV light to catalyze the decomposition of H2O2. The introduction of UV irradiation to the Fenton system largely increased the degradation rate of HCHO, mainly as a result of the accelerating effect on the formation of the Fe2+/Fe3+ cycle. The reaction products were analyzed by gas chromatography-mass spectrometry and a chemical oxygen demand (COD) analyzer. The effluent gases also were analyzed by gas chromatography. Based on those results, the reaction pathways of HCHO in the UV-Fenton system were proposed. The qualitative and quantitative analysis of the reaction products and the COD showed that the main intermediate product of the reaction was formic acid, and the further oxidation of it was the rate-limiting step for the degradation of HCHO.

  11. Genomic organisation, activity and distribution analysis of the microbial putrescine oxidase degradation pathway.

    Science.gov (United States)

    Foster, Alexander; Barnes, Nicole; Speight, Robert; Keane, Mark A

    2013-10-01

    The catalytic action of putrescine specific amine oxidases acting in tandem with 4-aminobutyraldehyde dehydrogenase is explored as a degradative pathway in Rhodococcus opacus. By limiting the nitrogen source, increased catalytic activity was induced leading to a coordinated response in the oxidative deamination of putrescine to 4-aminobutyraldehyde and subsequent dehydrogenation to 4-aminobutyrate. Isolating the dehydrogenase by ion exchange chromatography and gel filtration revealed that the enzyme acts principally on linear aliphatic aldehydes possessing an amino moiety. Michaelis-Menten kinetic analysis delivered a Michaelis constant (K(M)=0.014 mM) and maximum rate (Vmax=11.2 μmol/min/mg) for the conversion of 4-aminobutyraldehyde to 4-aminobutyrate. The dehydrogenase identified by MALDI-TOF mass spectrometric analysis (E value=0.031, 23% coverage) belongs to a functionally related genomic cluster that includes the amine oxidase, suggesting their association in a directed cell response. Key regulatory, stress and transport encoding genes have been identified, along with candidate dehydrogenases and transaminases for the further conversion of 4-aminobutyrate to succinate. Genomic analysis has revealed highly similar metabolic gene clustering among members of Actinobacteria, providing insight into putrescine degradation notably among Micrococcaceae, Rhodococci and Corynebacterium by a pathway that was previously uncharacterised in bacteria.

  12. Pathways and Determinants of Early Spontaneous Vegetation Succession in Degraded Lowland of South China

    Institute of Scientific and Technical Information of China (English)

    Wen-Jun Duan; Hai Ran; Sheng-Lei Fu; Qin-Feng Guo; Jun Wang

    2008-01-01

    Continuous and prolonged human disturbances have caused severe degradation of a large portion of lowland in South China, and how to restore such degraded ecosystems becomes an increasing concern. The process and mechanisms of spontaneous succession, which plays an important role in vegetation restoration, have not been adequately examined. To identify the pathways of early spontaneous vegetation succession, 41 plots representing plant communities abandoned over different times were established and Investigated. The communities and indicator species of the vegetation were classified by analyzing the important values of plant species using multivariate analyses. The reaults indicated that the plant species could be classified into nine plant communities repreaenting six succession staages. The pathway and species composition alao changed in the process of succession. We also meaeurad 13 environmental variables of microtopography, soil structure and soil nutrition in each plot to examine the driving forces of auccession and the vegetation-environment relationships. Our resulta ahowed that the environmental variables changed in diverse directions, and that aoil bulk density, soil water capacity and soU acidity were the most important factors.

  13. Degradation Pathway for Eplerenone by Validated Stability Indicating UP-LC Method.

    Science.gov (United States)

    Sudhakar Babu, Kondru; Madireddy, Venkataramanna; Indukuri, Venkata Somaraju

    2012-01-01

    Degradation pathway for eplerenone is established as per ICH recommendations by validated and stability-indicating reverse phase liquid chromatographic method. Eplerenone is subjected to stress conditions of acid, base, oxidation, and thermal and photolysis. Significant degradation is observed in acid and base stress conditions. Four impurities are studied and the major degradant (RRT about 0.31) was identified by LC-MS and spectral analysis. The stress samples are assayed against a qualified reference standard and the mass balance is found close to 99.5%. Efficient chromatographic separation is achieved on a Waters symmetry C18 stationary phase with simple mobile phase combination delivered in gradient mode and quantification is carried at 240 nm at a flow rate of 1.0 mL min(-1). In the developed LC method the resolution between eplerenone and four potential impurities (imp-1, imp-2, imp-3, and imp-4) is found to be greater than 4.0. Regression analysis shows an r value (correlation coefficient) of greater than 0.999 for eplerenone and four potential impurities. This method is capable to detect the impurities of eplerenone at a level of 0.020% with respect to test concentration of 1.0 mg mL(-1) for a 20 μL injection volume. The developed UPLC method is validated with respect to specificity, linearity and range, accuracy, precision, and robustness for impurities and assay determination.

  14. Autophagy-lysosomal pathway is involved in lipid degradation in rat liver.

    Science.gov (United States)

    Skop, V; Cahová, M; Papáčková, Z; Páleníčková, E; Daňková, H; Baranowski, M; Zabielski, P; Zdychová, J; Zídková, J; Kazdová, L

    2012-01-01

    We present data supporting the hypothesis that the lysosomal-autophagy pathway is involved in the degradation of intracellular triacylglycerols in the liver. In primary hepatocytes cultivated in the absence of exogenous fatty acids (FFA), both inhibition of autophagy flux (asparagine) or lysosomal activity (chloroquine) decreased secretion of VLDL (very low density lipoproteins) and formation of FFA oxidative products while the stimulation of autophagy by rapamycine increased some of these parameters. Effect of rapamycine was completely abolished by inactivation of lysosomes. Similarly, when autophagic activity was influenced by cultivating the hepatocytes in "starving" (amino-acid poor medium) or "fed" (serum-supplemented medium) conditions, VLDL secretion and FFA oxidation mirrored the changes in autophagy being higher in starvation and lower in fed state. Autophagy inhibition as well as lysosomal inactivation depressed FFA and DAG (diacylglycerol) formation in liver slices in vitro. In vivo, intensity of lysosomal lipid degradation depends on the formation of autophagolysosomes, i.e. structures bringing the substrate for degradation and lysosomal enzymes into contact. We demonstrated that lysosomal lipase (LAL) activity in liver autophagolysosomal fraction was up-regulated in fasting and down-regulated in fed state together with the increased translocation of LAL and LAMP2 proteins from lysosomal pool to this fraction. Changes in autophagy intensity (LC3-II/LC3-I ratio) followed a similar pattern.

  15. Photocatalytic transformation of sixteen substituted phenylurea herbicides in aqueous semiconductor suspensions: intermediates and degradation pathways.

    Science.gov (United States)

    Fenoll, José; Sabater, Paula; Navarro, Gines; Pérez-Lucas, Gabriel; Navarro, Simón

    2013-01-15

    The photocatalytic degradation of sixteen substituted phenylurea herbicides (PUHs) in pure water has been studied using zinc oxide (ZnO) and titanium dioxide (TiO(2)) as photocatalyst under artificial light irradiation. Photocatalytic experiments showed that the addition of these chalcogenide oxides in tandem with the oxidant (Na(2)S(2)O(8)) strongly enhances the degradation rate of these compounds in comparison with those carried out with ZnO and TiO(2) alone and photolytic tests. Comparison of catalysts showed that ZnO is the most efficient for the removal of such herbicides in optimal conditions and at constant volumetric rate of photon absorption in the photoreactor. Thus, the complete disappearance of all the studied compounds was achieved after 20 min of illumination in the ZnO/Na(2)S(2)O(8) system. The main photocatalytic intermediates detected during the degradation of PUHs were identified. The probable photodegradation pathways were proposed and discussed. The main steps involved: N-demethylation of the N,N-dimethylurea-substituted compounds followed of N-demethylation and N-demethoxylation of the N-methoxy-N-methyl-substituted ureas and hydroxylation of aromatic rings and their aliphatic side-chains of both, parent compounds and intermediates.

  16. WNK4 enhances the degradation of NCC through a sortilin-mediated lysosomal pathway.

    Science.gov (United States)

    Zhou, Bo; Zhuang, Jieqiu; Gu, Dingying; Wang, Hua; Cebotaru, Liudmila; Guggino, William B; Cai, Hui

    2010-01-01

    WNK kinase is a serine/threonine kinase that plays an important role in electrolyte homeostasis. WNK4 significantly inhibits the surface expression of the sodium chloride co-transporter (NCC) by enhancing the degradation of NCC through a lysosomal pathway, but the mechanisms underlying this trafficking are unknown. Here, we investigated the effect of the lysosomal targeting receptor sortilin on NCC expression and degradation. In Cos-7 cells, we observed that the presence of WNK4 reduced the steady-state amount of NCC by approximately half. Co-transfection with truncated sortilin (a dominant negative mutant) prevented this WNK4-induced reduction in NCC. NCC immunoprecipitated with both wild-type sortilin and, to a lesser extent, truncated sortilin. Immunostaining revealed that WNK4 increased the co-localization of NCC with the lysosomal marker cathepsin D, and NCC co-localized with wild-type sortilin, truncated sortilin, and WNK4 in the perinuclear region. These findings suggest that WNK4 promotes NCC targeting to the lysosome for degradation via a mechanism involving sortilin.

  17. Unfolded protein response and activated degradative pathways regulation in GNE myopathy.

    Directory of Open Access Journals (Sweden)

    Honghao Li

    Full Text Available Although intracellular beta amyloid (Aβ accumulation is known as an early upstream event in the degenerative course of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE myopathy, the process by which Aβdeposits initiate various degradative pathways, and their relationship have not been fully clarified. We studied the possible secondary responses after amyloid beta precursor protein (AβPP deposition including unfolded protein response (UPR, ubiquitin proteasome system (UPS activation and its correlation with autophagy system. Eight GNE myopathy patients and five individuals with normal muscle morphology were included in this study. We performed immunofluorescence and immunoblotting to investigate the expression of AβPP, phosphorylated tau (p-tau and endoplasmic reticulum molecular chaperones. Proteasome activities were measured by cleavage of fluorogenic substrates. The expression of proteasome subunits and linkers between proteasomal and autophagy systems were also evaluated by immunoblotting and relative quantitative real-time RT-PCR. Four molecular chaperones, glucose-regulated protein 94 (GRP94, glucose-regulated protein 78 (GRP78, calreticulin and calnexin and valosin containing protein (VCP were highly expressed in GNE myopathy. 20S proteasome subunits, three main proteasome proteolytic activities, and the factors linking UPS and autophagy system were also increased. Our study suggests that AβPP deposition results in endoplasmic reticulum stress (ERS and highly expressed VCP deliver unfolded proteins from endoplasmic reticulum to proteosomal system which is activated in endoplasmic reticulum associated degradation (ERAD in GNE myopathy. Excessive ubiquitinated unfolded proteins are exported by proteins that connect UPS and autophagy to autophagy system, which is activated as an alternative pathway for degradation.

  18. Identification of genes and pathways related to phenol degradation in metagenomic libraries from petroleum refinery wastewater.

    Directory of Open Access Journals (Sweden)

    Cynthia C Silva

    Full Text Available Two fosmid libraries, totaling 13,200 clones, were obtained from bioreactor sludge of petroleum refinery wastewater treatment system. The library screening based on PCR and biological activity assays revealed more than 400 positive clones for phenol degradation. From these, 100 clones were randomly selected for pyrosequencing in order to evaluate the genetic potential of the microorganisms present in wastewater treatment plant for biodegradation, focusing mainly on novel genes and pathways of phenol and aromatic compound degradation. The sequence analysis of selected clones yielded 129,635 reads at an estimated 17-fold coverage. The phylogenetic analysis showed Burkholderiales and Rhodocyclales as the most abundant orders among the selected fosmid clones. The MG-RAST analysis revealed a broad metabolic profile with important functions for wastewater treatment, including metabolism of aromatic compounds, nitrogen, sulphur and phosphorus. The predicted 2,276 proteins included phenol hydroxylases and cathecol 2,3- dioxygenases, involved in the catabolism of aromatic compounds, such as phenol, byphenol, benzoate and phenylpropanoid. The sequencing of one fosmid insert of 33 kb unraveled the gene that permitted the host, Escherichia coli EPI300, to grow in the presence of aromatic compounds. Additionally, the comparison of the whole fosmid sequence against bacterial genomes deposited in GenBank showed that about 90% of sequence showed no identity to known sequences of Proteobacteria deposited in the NCBI database. This study surveyed the functional potential of fosmid clones for aromatic compound degradation and contributed to our knowledge of the biodegradative capacity and pathways of microbial assemblages present in refinery wastewater treatment system.

  19. Identification of genes and pathways related to phenol degradation in metagenomic libraries from petroleum refinery wastewater.

    Science.gov (United States)

    Silva, Cynthia C; Hayden, Helen; Sawbridge, Tim; Mele, Pauline; De Paula, Sérgio O; Silva, Lívia C F; Vidigal, Pedro M P; Vicentini, Renato; Sousa, Maíra P; Torres, Ana Paula R; Santiago, Vânia M J; Oliveira, Valéria M

    2013-01-01

    Two fosmid libraries, totaling 13,200 clones, were obtained from bioreactor sludge of petroleum refinery wastewater treatment system. The library screening based on PCR and biological activity assays revealed more than 400 positive clones for phenol degradation. From these, 100 clones were randomly selected for pyrosequencing in order to evaluate the genetic potential of the microorganisms present in wastewater treatment plant for biodegradation, focusing mainly on novel genes and pathways of phenol and aromatic compound degradation. The sequence analysis of selected clones yielded 129,635 reads at an estimated 17-fold coverage. The phylogenetic analysis showed Burkholderiales and Rhodocyclales as the most abundant orders among the selected fosmid clones. The MG-RAST analysis revealed a broad metabolic profile with important functions for wastewater treatment, including metabolism of aromatic compounds, nitrogen, sulphur and phosphorus. The predicted 2,276 proteins included phenol hydroxylases and cathecol 2,3- dioxygenases, involved in the catabolism of aromatic compounds, such as phenol, byphenol, benzoate and phenylpropanoid. The sequencing of one fosmid insert of 33 kb unraveled the gene that permitted the host, Escherichia coli EPI300, to grow in the presence of aromatic compounds. Additionally, the comparison of the whole fosmid sequence against bacterial genomes deposited in GenBank showed that about 90% of sequence showed no identity to known sequences of Proteobacteria deposited in the NCBI database. This study surveyed the functional potential of fosmid clones for aromatic compound degradation and contributed to our knowledge of the biodegradative capacity and pathways of microbial assemblages present in refinery wastewater treatment system.

  20. Characterization of Starch Degradation Related Genes in Postharvest Kiwifruit.

    Science.gov (United States)

    Hu, Xiong; Kuang, Sheng; Zhang, Ai-Di; Zhang, Wang-Shu; Chen, Miao-Jin; Yin, Xue-Ren; Chen, Kun-Song

    2016-12-15

    Starch is one of the most important storage carbohydrates in plants. Kiwifruit typically accumulate large amounts of starch during development. The fruit retain starch until commercial maturity, and its postharvest degradation is essential for consumer acceptance. The activity of genes related to starch degradation has, however, rarely been investigated. Based on the kiwifruit genome sequence and previously reported starch degradation-related genes, 17 novel genes were isolated and the relationship between their expression and starch degradation was examined using two sets of materials: ethylene-treated (100 µL/L, 20 °C; ETH) vs. control (20 °C; CK) and controlled atmosphere stored (CA, 5% CO₂ + 2% O₂, 0 °C) vs. normal atmosphere in cold storage (NA, 0 °C). Physiological analysis indicated that ETH accelerated starch degradation and increased soluble solids content (SSC) and soluble sugars (glucose, fructose and sucrose), while CA inhibited starch reduction compared with NA. Using these materials, expression patterns of 24 genes that may contribute to starch degradation (seven previously reported and 17 newly isolated) were analyzed. Among the 24 genes, AdAMY1, AdAGL3 and AdBAM3.1/3L/9 were significantly induced by ETH and positively correlated with starch degradation. Furthermore, these five genes were also inhibited by CA, conforming the likely involvement of these genes in starch degradation. Thus, the present study has identified the genes with potential for involvement in starch degradation in postharvest kiwifruit, which will be useful for understanding the regulation of kiwifruit starch content and metabolism.

  1. Characterization of Starch Degradation Related Genes in Postharvest Kiwifruit

    Directory of Open Access Journals (Sweden)

    Xiong Hu

    2016-12-01

    Full Text Available Starch is one of the most important storage carbohydrates in plants. Kiwifruit typically accumulate large amounts of starch during development. The fruit retain starch until commercial maturity, and its postharvest degradation is essential for consumer acceptance. The activity of genes related to starch degradation has, however, rarely been investigated. Based on the kiwifruit genome sequence and previously reported starch degradation-related genes, 17 novel genes were isolated and the relationship between their expression and starch degradation was examined using two sets of materials: ethylene-treated (100 µL/L, 20 °C; ETH vs. control (20 °C; CK and controlled atmosphere stored (CA, 5% CO2 + 2% O2, 0 °C vs. normal atmosphere in cold storage (NA, 0 °C. Physiological analysis indicated that ETH accelerated starch degradation and increased soluble solids content (SSC and soluble sugars (glucose, fructose and sucrose, while CA inhibited starch reduction compared with NA. Using these materials, expression patterns of 24 genes that may contribute to starch degradation (seven previously reported and 17 newly isolated were analyzed. Among the 24 genes, AdAMY1, AdAGL3 and AdBAM3.1/3L/9 were significantly induced by ETH and positively correlated with starch degradation. Furthermore, these five genes were also inhibited by CA, conforming the likely involvement of these genes in starch degradation. Thus, the present study has identified the genes with potential for involvement in starch degradation in postharvest kiwifruit, which will be useful for understanding the regulation of kiwifruit starch content and metabolism.

  2. Characterization of Starch Degradation Related Genes in Postharvest Kiwifruit

    Science.gov (United States)

    Hu, Xiong; Kuang, Sheng; Zhang, Ai-Di; Zhang, Wang-Shu; Chen, Miao-Jin; Yin, Xue-Ren; Chen, Kun-Song

    2016-01-01

    Starch is one of the most important storage carbohydrates in plants. Kiwifruit typically accumulate large amounts of starch during development. The fruit retain starch until commercial maturity, and its postharvest degradation is essential for consumer acceptance. The activity of genes related to starch degradation has, however, rarely been investigated. Based on the kiwifruit genome sequence and previously reported starch degradation-related genes, 17 novel genes were isolated and the relationship between their expression and starch degradation was examined using two sets of materials: ethylene-treated (100 µL/L, 20 °C; ETH) vs. control (20 °C; CK) and controlled atmosphere stored (CA, 5% CO2 + 2% O2, 0 °C) vs. normal atmosphere in cold storage (NA, 0 °C). Physiological analysis indicated that ETH accelerated starch degradation and increased soluble solids content (SSC) and soluble sugars (glucose, fructose and sucrose), while CA inhibited starch reduction compared with NA. Using these materials, expression patterns of 24 genes that may contribute to starch degradation (seven previously reported and 17 newly isolated) were analyzed. Among the 24 genes, AdAMY1, AdAGL3 and AdBAM3.1/3L/9 were significantly induced by ETH and positively correlated with starch degradation. Furthermore, these five genes were also inhibited by CA, conforming the likely involvement of these genes in starch degradation. Thus, the present study has identified the genes with potential for involvement in starch degradation in postharvest kiwifruit, which will be useful for understanding the regulation of kiwifruit starch content and metabolism. PMID:27983700

  3. Synthesis and degradation pathways, functions, and pathology of ceramides and epidermal acylceramides.

    Science.gov (United States)

    Kihara, Akio

    2016-07-01

    Ceramide (Cer) is a structural backbone of sphingolipids and is composed of a long-chain base and a fatty acid. Existence of a variety of Cer species, which differ in chain-length, hydroxylation status, and/or double bond number of either of their hydrophobic chains, has been reported. Ceramide is produced by Cer synthases. Mammals have six Cer synthases (CERS1-6), each of which exhibits characteristic substrate specificity toward acyl-CoAs with different chain-lengths. Knockout mice for each Cer synthase show corresponding, isozyme-specific phenotypes, revealing the functional differences of Cers with different chain-lengths. Cer diversity is especially prominent in epidermis. Changes in Cer levels, composition, and chain-lengths are associated with atopic dermatitis. Acylceramide (acyl-Cer) specifically exists in epidermis and plays an essential role in skin permeability barrier formation. Accordingly, defects in acyl-Cer synthesis cause the cutaneous disorder ichthyosis with accompanying severe skin barrier defects. Although the molecular mechanism by which acyl-Cer is generated was long unclear, most genes involved in its synthesis have been identified recently. In Cer degradation pathways, the long-chain base moiety of Cer is converted to acyl-CoA, which is then incorporated mainly into glycerophospholipids. This pathway generates the lipid mediator sphingosine 1-phosphate. This review will focus on recent advances in our understanding of the synthesis and degradation pathways, physiological functions, and pathology of Cers/acyl-Cers. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Photodegradation of gemfibrozil in aqueous solution under UV irradiation: kinetics, mechanism, toxicity, and degradation pathways.

    Science.gov (United States)

    Ma, Jingshuai; Lv, Wenying; Chen, Ping; Lu, Yida; Wang, Fengliang; Li, Fuhua; Yao, Kun; Liu, Guoguang

    2016-07-01

    The lipid regulator gemfibrozil (GEM) has been reported to be persistent in conventional wastewater treatment plants. This study investigated the photolytic behavior, toxicity of intermediate products, and degradation pathways of GEM in aqueous solutions under UV irradiation. The results demonstrated that the photodegradation of GEM followed pseudo-first-order kinetics, and the pseudo-first-order rate constant was decreased markedly with increasing initial concentrations of GEM and initial pH. The photodegradation of GEM included direct photolysis via (3)GEM(*) and self-sensitization via ROS, where the contribution rates of degradation were 0.52, 90.05, and 8.38 % for ·OH, (1)O2, and (3)GEM(*), respectively. Singlet oxygen ((1)O2) was evidenced by the molecular probe compound, furfuryl alcohol (FFA), and was identified as the primary reactive species in the photolytic process. The steady-state concentrations of (1)O2 increased from (0.324 ± 0.014) × 10(-12) to (1.021 ± 0.040) × 10(-12) mol L(-1), as the initial concentrations of GEM were increased from 5 to 20 mg L(-1). The second-order rate constant for the reaction of GEM with (1)O2 was calculated to be 2.55 × 10(6) M(-1) s(-1). The primary transformation products were identified using HPLC-MS/MS, and possible photodegradation pathways were proposed by hydroxylation, aldehydes reactions, as well as the cleavage of ether side chains. The toxicity of phototransformation product evaluation revealed that photolysis potentially provides a critical pathway for GEM toxicity reduction in potable water and wastewater treatment facilities.

  5. Stress-induced nuclear RNA degradation pathways regulate yeast bromodomain factor 2 to promote cell survival.

    Directory of Open Access Journals (Sweden)

    Kevin Roy

    2014-09-01

    Full Text Available Bromodomain proteins are key regulators of gene expression. How the levels of these factors are regulated in specific environmental conditions is unknown. Previous work has established that expression of yeast Bromodomain factor 2 (BDF2 is limited by spliceosome-mediated decay (SMD. Here we show that BDF2 is subject to an additional layer of post-transcriptional control through RNase III-mediated decay (RMD. We found that the yeast RNase III Rnt1p cleaves a stem-loop structure within the BDF2 mRNA to down-regulate its expression. However, these two nuclear RNA degradation pathways play distinct roles in the regulation of BDF2 expression, as we show that the RMD and SMD pathways of the BDF2 mRNA are differentially activated or repressed in specific environmental conditions. RMD is hyper-activated by salt stress and repressed by hydroxyurea-induced DNA damage while SMD is inactivated by salt stress and predominates during DNA damage. Mutations of cis-acting signals that control SMD and RMD rescue numerous growth defects of cells lacking Bdf1p, and show that SMD plays an important role in the DNA damage response. These results demonstrate that specific environmental conditions modulate nuclear RNA degradation pathways to control BDF2 expression and Bdf2p-mediated gene regulation. Moreover, these results show that precise dosage of Bromodomain factors is essential for cell survival in specific environmental conditions, emphasizing their importance for controlling chromatin structure and gene expression in response to environmental stress.

  6. Aqueous photodegradation of 4-tert-butylphenol: By-products, degradation pathway and theoretical calculation assessment

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Yanlin [State Key Laboratory of Pollution Control and Resources Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai 200092 (China); Shi, Jin; Chen, Hongche [Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention, Department of Environmental Science & Engineering, Fudan University, Shanghai 200433 (China); Zhao, Jianfu [State Key Laboratory of Pollution Control and Resources Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai 200092 (China); Dong, Wenbo, E-mail: wbdong@fudan.edu.cn [Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention, Department of Environmental Science & Engineering, Fudan University, Shanghai 200433 (China)

    2016-10-01

    4-tert-butylphenol (4-t-BP), an endocrine disrupting chemical, is widely distributed in natural bodies of water but is difficult to biodegrade. In this study, we focused on the transformation of 4-t-BP in photo-initiated degradation processes. The steady-state photolysis and laser flash photolysis (LFP) experiments were conducted in order to elucidate its degradation mechanism. Identification of products was performed using the GC–MS, LC-MS and theoretical calculation techniques. The oxidation pathway of 4-t-BP by hydroxyl radical (HO·) was also studied and H{sub 2}O{sub 2} was added to produce HO·. 4-tert-butylcatechol and 4-tert-butylphenol dimer were produced in 4-t-BP direct photolysis. 4-tert-butylcatechol and hydroquinone were produced by the oxidation of HO·. But the formation mechanism of 4-tert-butylcatechol in the two processes was different. The benzene ring was fractured in 4-t-BP oxidation process and 29% of TOC was degraded after 16 h irradiation. - Highlights: • Photodegradation of 4-t-BP, an endocrine disrupting chemical, has been investigated. • 3 stable byproducts were identified from photolysis and oxidation processes. • 5 transient by-products were concluded from LFP experiments. • The theoretical calculation was performed to confirm the byproducts. • 4-t-BP was degraded with increasing efficiency: 254 nm < H{sub 2}O{sub 2}/313 nm < H{sub 2}O{sub 2}/254 nm.

  7. Degradation Pathways for Geogenic Volatile Organic Compounds (VOCs) in Soil Gases from the Solfatara Crater (Campi Flegrei, Southern Italy).

    Science.gov (United States)

    Tassi, F.; Venturi, S.; Cabassi, J.; Capecchiacci, F.; Nisi, B., Sr.; Vaselli, O.

    2014-12-01

    The chemical composition of volatile organic compounds (VOCs) in soil gases from the Solfatara crater (Campi Flegrei, Southern Italy) was analyzed to investigate the effects of biogeochemical processes occurring within the crater soil on gases discharged from the hydrothermal reservoir and released into the atmosphere through diffuse degassing. In this system, two fumarolic vents (namely Bocca Grande and Bocca Nuova) are the preferential pathways for hydrothermal fluid uprising. For our goal, the chemistry of VOCs discharged from these sites were compared to that of soil gases. Our results highlighted that C4-C9 alkanes, alkenes, S-bearing compounds and alkylated aromatics produced at depth were the most prone to degradation processes, such as oxidation-reduction and hydration-dehydration reactions, as well as to microbial activity. Secondary products, which were enriched in sites characterized by low soil gas fluxes, mostly consisted of aldheydes, ketons, esters, ethers, organic acids and, subordinately, alcohols. Benzene, phenol and hydrofluorocarbons (HCFCs) produced at depth were able to transit through the soil almost undisturbed, independently on the emission rate of diffuse degassing. The presence of cyclics was possibly related to an independent low-temperature VOC source, likely within sedimentary formations overlying the hydrothermal reservoir. Chlorofluorocarbons (CFCs) were possibly due to air contamination. This study demonstrated the strict control of biogeochemical processes on the behaviour of hydrothermal VOCs that, at least at a local scale, may have a significant impact on air quality. Laboratory experiments conducted at specific chemical-physical conditions and in presence of different microbial populations may provide useful information for the reconstruction of the degradation pathways controlling fate and behaviour of VOCs in the soil.

  8. Isolation and characterization of engine oil degrading indigenous ...

    African Journals Online (AJOL)

    AJB SERVER

    2007-01-04

    Jan 4, 2007 ... bacterial isolates were responsible for the oil degradation. All isolates were ... and water soluble) hydrocarbons that would be more of a concern for ... Mechanical method to reduce hydrocarbon pollution is expensive and time ...

  9. The rheology, degradation, processing, and characterization of renewable resource polymers

    Science.gov (United States)

    Conrad, Jason David

    Renewable resource polymers have become an increasingly popular alternative to conventional fossil fuel based polymers over the past couple decades. The push by the government as well as both industrial and consumer markets to go "green" has provided the drive for companies to research and develop new materials that are more environmentally friendly and which are derived from renewable materials. Two polymers that are currently being produced commercially are poly-lactic acid (PLA) and polyhydroxyalkanoate (PHA) copolymers, both of which can be derived from renewable feedstocks and have shown to exhibit similar properties to conventional materials such as polypropylene, polyethylene, polystyrene, and PET. PLA and PHA are being used in many applications including food packaging, disposable cups, grocery bags, and biomedical applications. In this work, we report on the rheological properties of blends of PLA and PHA copolymers. The specific materials used in the study include Natureworks RTM 7000D grade PLA and PHA copolymers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Blends ranging from 10 to 50 percent PHA by weight are also examined. Shear and extensional experiments are performed to characterize the flow behavior of the materials in different flow fields. Transient experiments are performed to study the shear rheology over time in order to determine how the viscoelastic properties change under typical processing conditions and understand the thermal degradation behavior of the materials. For the blends, it is determined that increasing the PHA concentration in the blend results in a decrease in viscosity and increase in degradation. Models are fit to the viscosity of the blends using the pure material viscosities in order to be able to predict the behavior at a given blend composition. We also investigate the processability of these materials into films and examine the resultant properties of the cast films. The mechanical and thermal properties of the

  10. Bacterial degradation of benzoate: cross-regulation between aerobic and anaerobic pathways

    OpenAIRE

    2012-01-01

    We have studied for the first time the transcriptional regulatory circuit that controls the expression of the box genes encoding the aerobic hybrid pathway used to assimilate benzoate via coenzyme A (CoA) derivatives in bacteria. The promoters responsible for the expression of the box cluster in the β-proteobacterium Azoarcus sp., their cognate transcriptional repressor, the BoxR protein, and the inducer molecule (benzoyl-CoA) have been characterized. The BoxR protein shows a significant sequ...

  11. Degradation of the synthetic dye amaranth by the fungus Bjerkandera adusta Dec 1: inference of the degradation pathway from an analysis of decolorized products.

    Science.gov (United States)

    Gomi, Nichina; Yoshida, Shuji; Matsumoto, Kazutsugu; Okudomi, Masayuki; Konno, Hiroki; Hisabori, Toru; Sugano, Yasushi

    2011-11-01

    We examined the degradation of amaranth, a representative azo dye, by Bjerkandera adusta Dec 1. The degradation products were analyzed by high performance liquid chromatography (HPLC), visible absorbance, and electrospray ionization time-of-flight mass spectroscopy (ESI-TOF-MS). At the primary culture stage (3 days), the probable reaction intermediates were 1-aminonaphthalene-2,3,6-triol, 4-(hydroxyamino) naphthalene-1-ol, and 2-hydroxy-3-[2-(4-sulfophenyl) hydrazinyl] benzenesulfonic acid. After 10 days, the reaction products detected were 4-nitrophenol, phenol, 2-hydroxy-3-nitrobenzenesulfonic acid, 4-nitrobenzene sulfonic acid, and 3,4'-disulfonyl azo benzene, suggesting that no aromatic amines were created. Manganese-dependent peroxidase activity increased sharply after 3 days culture. Based on these results, we herein propose, for the first time, a degradation pathway for amaranth. Our results suggest that Dec 1 degrades amaranth via the combined activities of peroxidase and hydrolase and reductase action.

  12. Sulfoacetate is degraded via a novel pathway involving sulfoacetyl-CoA and sulfoacetaldehyde in Cupriavidus necator H16.

    Science.gov (United States)

    Weinitschke, Sonja; Hollemeyer, Klaus; Kusian, Bernhard; Bowien, Botho; Smits, Theo H M; Cook, Alasdair M

    2010-11-12

    Bacterial degradation of sulfoacetate, a widespread natural product, proceeds via sulfoacetaldehyde and requires a considerable initial energy input. Whereas the fate of sulfoacetaldehyde in Cupriavidus necator (Ralstonia eutropha) H16 is known, the pathway from sulfoacetate to sulfoacetaldehyde is not. The genome sequence of the organism enabled us to hypothesize that the inducible pathway, which initiates sau (sulfoacetate utilization), involved a four-gene cluster (sauRSTU; H16_A2746 to H16_A2749). The sauR gene, divergently orientated to the other three genes, probably encodes the transcriptional regulator of the presumed sauSTU operon, which is subject to inducible transcription. SauU was tentatively identified as a transporter of the major facilitator superfamily, and SauT was deduced to be a sulfoacetate-CoA ligase. SauT was a labile protein, but it could be separated and shown to generate AMP and an unknown, labile CoA-derivative from sulfoacetate, CoA, and ATP. This unknown compound, analyzed by MALDI-TOF-MS, had a relative molecular mass of 889.7, which identified it as protonated sulfoacetyl-CoA (calculated 889.6). SauS was deduced to be sulfoacetaldehyde dehydrogenase (acylating). The enzyme was purified 175-fold to homogeneity and characterized. Peptide mass fingerprinting confirmed the sauS locus (H16_A2747). SauS converted sulfoacetyl-CoA and NADPH to sulfoacetaldehyde, CoA, and NADP(+), thus confirming the hypothesis.

  13. Understanding the degradation pathway of the pesticide, chlorpyrifos by noble metal nanoparticles.

    Science.gov (United States)

    Bootharaju, M S; Pradeep, T

    2012-02-01

    Application of nanoparticles (NPs) in environmental remediation such as water purification requires a detailed understanding of the mechanistic aspects of the interaction between the species involved. Here, an attempt was made to understand the chemistry of noble metal nanoparticle-pesticide interaction, as these nanosystems are being used extensively for water purification. Our model pesticide, chlorpyrifos (CP), belonging to the organophosphorothioate group, is shown to decompose to 3,5,6-trichloro-2-pyridinol (TCP) and diethyl thiophosphate at room temperature over Ag and Au NPs, in supported and unsupported forms. The degradation products were characterized by absorption spectroscopy and electrospray ionization mass spectrometry (ESI MS). These were further confirmed by ESI tandem mass spectrometry. The interaction of CP with NP surfaces was investigated using transmission electron microscopy, energy dispersive analysis of X-rays, Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS). XPS reveals no change in the oxidation state of silver after the degradation of CP. It is proposed that the degradation of CP proceeds through the formation of AgNP-S surface complex, which is confirmed by Raman spectroscopy. In this complex, the P-O bond cleaves to yield a stable aromatic species, TCP. The rate of degradation of CP increases with increase of temperature and pH. Complete degradation of 10 mL of 2 ppm CP solution is achieved in 3 h using 100 mg of supported Ag@citrate NPs on neutral alumina at room temperature at a loading of ∼0.5 wt %. The effect of alumina and monolayer protection of NPs on the degradation of CP is also investigated. The rate of degradation of CP by Ag NPs is greater than that of Au NPs. The results have implications to the application of noble metal NPs for drinking water purification, as pesticide contamination is prevalent in many parts of the world. Study shows that supported Ag and Au NPs may be employed in sustainable

  14. Removal and Degradation Pathways of Sulfamethoxazole Present in Synthetic Municipal Wastewater via an Anaerobic Membrane Bioreactor

    KAUST Repository

    Sanchez Huerta, Claudia

    2016-05-01

    The current global water crisis in addition to continues contamination of natural water bodies with harmful organic micropollutants (OMPs) have driven the development of new water treatment technologies that allow the efficient removal of such compounds. Among a long list of OMPs, antibiotics are considered as top priority pollutants to be treated due to their great resistance to biological treatments and their potential to develop bacterial resistance. Different approaches, such as membrane-based and advance oxidation processes have been proposed to alleviate or minimize antibiotics discharge into aquatic environments. However most of these processes are costly and generate either matrices with high concentration of OMPs or intermediate products with potentially greater toxicity or persistence. Therefore, this thesis proposes the study of an anaerobic membrane bioreactor (AnMBR) for the treatment of synthetic municipal wastewater containing sulfamethoxazole (SMX), a world widely used antibiotic. Besides the general evaluation of AnMBR performance in the COD removal and biogas production, this research mainly focuses on the SMX removal and its degradation pathway. Thus 5 SMX quantification was performed through solid phase extraction-liquid chromatography/mass spectrometry and the identification of its transformation products (TPs) was assessed by gas chromatography/mass spectrometry technique. The results achieved showed that, working under optimal conditions (35°C, pH 7 and ORP around -380 to -420 mV) and after a biomass adaptation period (maintaining 0.85 VSS/TSS ratio), the AnMBR process provided over 95% COD removal and 95-98% SMX removal, while allowing stable biogas composition and methane production (≈130 mL CH4/g CODremoved). Kinetic analysis through a batch test showed that after 24 h of biological reaction, AnMBR process achieved around 94% SMX removal, indicating a first order kinetic reaction with K= 0.119, which highlights the high degradation

  15. Photocatalytic transformation of sixteen substituted phenylurea herbicides in aqueous semiconductor suspensions: Intermediates and degradation pathways

    Energy Technology Data Exchange (ETDEWEB)

    Fenoll, José, E-mail: jose.fenoll@carm.es [Departamento de Calidad y Garantía Alimentaria, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), C/Mayor s/n, La Alberca, 30150 Murcia (Spain); Sabater, Paula [Departamento de Calidad y Garantía Alimentaria, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), C/Mayor s/n, La Alberca, 30150 Murcia (Spain); Navarro, Gines; Pérez-Lucas, Gabriel; Navarro, Simón [Departamento de Química Agrícola, Geología y Edafología, Facultad de Química, Universidad de Murcia, Campus Universitario de Espinardo, 30100 Murcia (Spain)

    2013-01-15

    Highlights: ► Photocatalytic oxidation of phenylurea herbicides (PUHs) in water. ► The study was performed using ZnO and TiO{sub 2} under artificial light irradiation. ► PUHs were totally degraded using ZnO/Na{sub 2}S{sub 2}O{sub 8}, ZnO, TiO{sub 2}/Na{sub 2}S{sub 2}O{sub 8} and TiO{sub 2}. ► ZnO is the most efficient photocatalyst for the removal of these herbicides. ► 13 intermediates were identified and a mechanism of degradation has been proposed. -- Abstract: The photocatalytic degradation of sixteen substituted phenylurea herbicides (PUHs) in pure water has been studied using zinc oxide (ZnO) and titanium dioxide (TiO{sub 2}) as photocatalyst under artificial light irradiation. Photocatalytic experiments showed that the addition of these chalcogenide oxides in tandem with the oxidant (Na{sub 2}S{sub 2}O{sub 8}) strongly enhances the degradation rate of these compounds in comparison with those carried out with ZnO and TiO{sub 2} alone and photolytic tests. Comparison of catalysts showed that ZnO is the most efficient for the removal of such herbicides in optimal conditions and at constant volumetric rate of photon absorption in the photoreactor. Thus, the complete disappearance of all the studied compounds was achieved after 20 min of illumination in the ZnO/Na{sub 2}S{sub 2}O{sub 8} system. The main photocatalytic intermediates detected during the degradation of PUHs were identified. The probable photodegradation pathways were proposed and discussed. The main steps involved: N-demethylation of the N,N-dimethylurea-substituted compounds followed of N-demethylation and N-demethoxylation of the N-methoxy-N-methyl-substituted ureas and hydroxylation of aromatic rings and their aliphatic side-chains of both, parent compounds and intermediates.

  16. The Degradation Pathway of the Mitophagy Receptor Atg32 Is Re-Routed by a Posttranslational Modification.

    Science.gov (United States)

    Levchenko, Mariia; Lorenzi, Isotta; Dudek, Jan

    2016-01-01

    The outer mitochondrial membrane protein Atg32 is the central receptor for mitophagy, the mitochondria-specific form of autophagy. Atg32 is an unstable protein, and is rapidly degraded under conditions in which mitophagy is not induced. Here we show that Atg32 undergoes a posttranslational modification upon induction of mitophagy. The modification is dependent on the core autophagic machinery, including Atg8, and on the mitophagy-specific adaptor protein Atg11. The modified Atg32 is targeted to the vacuole where it becomes stabilized when vacuolar proteases are deficient. Interestingly, we find that this degradation pathway differs from the degradation pathway of non-modified Atg32, which neither involves vacuolar proteases, nor the proteasome. These analyses reveal that a posttranslational modification discriminates a form of Atg32 targeting mitochondria for mitophagy from that, which escapes mitophagy by rapid degradation.

  17. Photocatalytic degradation of metoprolol tartrate in suspensions of two TiO{sub 2}-based photocatalysts with different surface area. Identification of intermediates and proposal of degradation pathways

    Energy Technology Data Exchange (ETDEWEB)

    Abramovic, Biljana, E-mail: biljana.abramovic@dh.uns.ac.rs [Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences, University of Novi Sad, Trg D. Obradovica 3, 21000 Novi Sad (Serbia); Kler, Sanja, E-mail: sanja.kler@dh.uns.ac.rs [Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences, University of Novi Sad, Trg D. Obradovica 3, 21000 Novi Sad (Serbia); Sojic, Daniela, E-mail: daniela.sojic@dh.uns.ac.rs [Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences, University of Novi Sad, Trg D. Obradovica 3, 21000 Novi Sad (Serbia); Lausevic, Mila, E-mail: milal@tmf.bg.ac.rs [Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11120 Belgrade (Serbia); Radovic, Tanja, E-mail: tradovic@tmf.bg.ac.rs [Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11120 Belgrade (Serbia); Vione, Davide, E-mail: davide.vione@unito.it [Dipartimento di Chimica Analitica, Universita di Torino, Via Pietro Giuria 5, 10125 Torino (Italy)

    2011-12-30

    Highlights: Black-Right-Pointing-Pointer Kinetics and efficiency of photocatalytic degradation of the {beta}{sub 1}-blocker metoprolol tartrate (MET). Black-Right-Pointing-Pointer Two TiO{sub 2} specimens employed. Black-Right-Pointing-Pointer Faster degradation of MET, but slower mineralization, obtained with the TiO{sub 2} specimen having lower surface area. Black-Right-Pointing-Pointer Photocatalytic transformation pathways of MET including mineralization. - Abstract: This study investigates the efficiency of the photocatalytic degradation of metoprolol tartrate (MET), a widely used {beta}{sub 1}-blocker, in TiO{sub 2} suspensions of Wackherr's 'Oxyde de titane standard' and Degussa P25. The study encompasses transformation kinetics and efficiency, identification of intermediates and reaction pathways. In the investigated range of initial concentrations (0.01-0.1 mM), the photocatalytic degradation of MET in the first stage of the reaction followed approximately a pseudo-first order kinetics. The TiO{sub 2} Wackherr induced a significantly faster MET degradation compared to TiO{sub 2} Degussa P25 when relatively high substrate concentrations were used. By examining the effect of ethanol as a scavenger of hydroxyl radicals ({center_dot}OH), it was shown that the reaction with {center_dot}OH played the main role in the photocatalytic degradation of MET. After 240 min of irradiation the reaction intermediates were almost completely mineralized to CO{sub 2} and H{sub 2}O, while the nitrogen was predominantly present as NH{sub 4}{sup +}. Reaction intermediates were studied in detail and a number of them were identified using LC-MS/MS (ESI+), which allowed the proposal of a tentative pathway for the photocatalytic transformation of MET as a function of the TiO{sub 2} specimen.

  18. M2-like macrophages are responsible for collagen degradation through a mannose receptor-mediated pathway

    DEFF Research Database (Denmark)

    Madsen, Daniel H; Leonard, Daniel; Masedunskas, Andrius

    2013-01-01

    of the collagen receptors mannose receptor (Mrc1) and urokinase plasminogen activator receptor-associated protein (Endo180 and Mrc2) impaired this intracellular collagen degradation pathway. This study demonstrates the importance of receptor-mediated cellular uptake to collagen turnover in vivo and identifies......Tissue remodeling processes critically depend on the timely removal and remodeling of preexisting collagen scaffolds. Nevertheless, many aspects related to the turnover of this abundant extracellular matrix component in vivo are still incompletely understood. We therefore took advantage of recent...... advances in optical imaging to develop an assay to visualize collagen turnover in situ and identify cell types and molecules involved in this process. Collagen introduced into the dermis of mice underwent cellular endocytosis in a partially matrix metalloproteinase-dependent manner and was subsequently...

  19. [Isolation identification and characterization of halotolerant petroleum-degrading bacteria].

    Science.gov (United States)

    Wu, Tao; Xie, Wen-Jun; Yi, Yan-Li; Li, Xiao-Bin; Wang, Jun; Hu, Xiang-Ming

    2012-11-01

    To obtain efficient halotolerant petroleum-degrading bacteria, 39 bacteria strains were isolated from 30 petroleum contaminated saline soil samples in Yellow River Delta, an important base of petroleum production in China. One bacterium (strain BM38) was found to efficiently degrade crude oil in highly saline environments based on a series of liquid and soil incubation experiments. According to its morphology, physiochemical characteristics and 16S rDNA sequence analysis, this strain was identified as Pseudomonas putida. Moreover, a series of liquid incubation experiments were conducted to investigate its characteristics such as halotolerance, biosurfactants production and degrading efficiency for various hydrocarbons. The salt resistance test demonstrated that strain BM38 grew well at NaCl concentrations ranging from 0.5% to 6.0%. Petroleum degradation experiments showed that strain BM38 could degrade 73.5% crude oil after 7 days in a liquid culture medium containing 1.0% NaCl and remove more than 40% of total petroleum hydrocarbons after 40 days in the soil with 0.22% and 0.61% of salinity, these results proved that the strain was effective in removing petroleum hydrocarbons. Strain BM38 could produce a bioemulsifier in a liquid culture medium. The NaCl concentration had the significant effect on the EI24 of fermentation broth, which decreased sharply if the NaCl concentration was greater than 1.0%. However, the EI24 of BM38 was still quite high in the presence of 2.0% of NaCl, and the value was 61.0%. Furthermore, this strain was also able to grow in mineral liquid media amended with hexadecane, toluene, phenanthrene, isooctane and cyclohexane as the sole carbon sources. Among these hydracarbons, strain BM38 showed relatively high ability in degrading n-alkanes and aromatic hydracarbons. The results indicated that strain BM38 had potential for application in bioremediation of petroleum-contaminated saline soil.

  20. Isolation and characterization of an SDS-degrading Klebsiella oxytoca.

    Science.gov (United States)

    Shukor, M Y; Husin, W S W; Rahman, M F A; Shamaan, N A; Syed, M A

    2009-01-01

    Sodium dodecyl sulfate (SDS) is one of the main components in the detergent and cosmetic industries. Its bioremediation by suitable microorganism has begun to receive greater attention as the amount of SDS usage increases to a point where treatment plants would not be able to cope with the increasing amount of SDS in wastewater. The purpose of this work was to isolate local SDS-degrading bacteria. Screening was carried out by the conventional enrichment-culture technique. Six SDS-degrading bacteria were isolated. Of these isolates, isolate S14 showed the highest degradation of SDS with 90% degradation after three days of incubation. Isolate S14 was tentatively identified as Klebsiella oxytoca strain DRY14 based on carbon utilization profiles using Biolog GN plates and partial 16S rDNA molecular phylogeny. SDS degradation by the bacterium was optimum at 37 degrees 0. Ammonium sulphate; at 2.0 g l(-1), was found to be the best nitrogen source for the growth of strain DRY14. Maximum growth on SDS was observed at pH 7.25. The strain exhibited optimum growth at SDS concentration of 2.0 g l(-1) and was completely inhibited at 10 g l(-1) SDS. At the tolerable initial concentration of 2.0 g l(-1), almost 80% of 2.0 g l(-1) SDS was degraded after 4 days of incubation concomitant with increase in cellular growth. The K(m(app) and V(max(app)) values calculated for the alkylsulfatase from this bacterium were 0.1 mM SDS and 1.07 micromol min(-1) mg(-1) protein, respectively.

  1. Isolation of a Pseudomonas Stutzeri strain that degrades1, 2, 4-trichlorobenzene and characterization of its degradative plasmid

    Institute of Scientific and Technical Information of China (English)

    Lei SONG; Hui WANG; Hanchang SHI; Hongying HU

    2008-01-01

    The genetic information encoding metabolic pathways for xenobiotic compounds in bacteria often resides on catabolic plasmids. The aim of the present work was to know the location of the genes for degrading 1, 2, 4-trichlorobenzen. In this paper a 1, 2, 4-trichlorobenzene-degrading strain THSL-1 was isolated from the soil of Tianjin Chemical Plant using 1, 2, 4-trichlorobenzene as the sole carbon source. The strain was identified as Pseudomonas stutzeri through morphologic survey and 16S rDNA sequence determination. A plasmid was discovered from strain THSL-1 by using the alkali lysis method. When the plasmid was transformed into E. coli. JM109 by the CaCl2 method, the transformant could grow using 1, 2, 4-trichlorobenzene as the sole carbon source and had the degradation function of 1, 2, 4-trichlorobenzene. Therefore, it could be deemed that the plasmid carried the degradative genes of 1, 2, 4-trichlorobenzene. The average size of the plasmid was finally determined to be 40.2 Kb using selectively three kinds of restricted inscribed enzymes (HindIII, BamHI, and XholI) for single cutting and double cutting the plasmid pTHSL-1, respectively.

  2. Degradation pathways of lamotrigine under advanced treatment by direct UV photolysis, hydroxyl radicals, and ozone.

    Science.gov (United States)

    Keen, Olya S; Ferrer, Imma; Michael Thurman, E; Linden, Karl G

    2014-12-01

    Lamotrigine is recently recognized as a persistent pharmaceutical in the water environment and wastewater effluents. Its degradation was studied under UV and ozone advanced oxidation treatments with reaction kinetics of lamotrigine with ozone (≈4 M(-1)s(-1)), hydroxyl radical [(2.1 ± 0.3) × 10(9)M(-1)s(-1)] and by UV photolysis with low and medium pressure mercury vapor lamps [quantum yields ≈0 and (2.7 ± 0.4)× 10(-4) respectively] determined. All constants were measured at pH 6 and at temperature ≈20°C. The results indicate that lamotrigine is slow to respond to direct photolysis or oxidation by ozone and no attenuation of the contaminant is expected in UV or ozone disinfection applications. The compound reacts rapidly with hydroxyl radicals indicating that advanced oxidation processes would be effective for its treatment. Degradation products were identified under each treatment process using accurate mass time-of-flight spectrometry and pathways of decay were proposed. The main transformation pathways in each process were: dechlorination of the benzene ring during direct photolysis; hydroxyl group addition to the benzene ring during the reaction with hydroxyl radicals; and triazine ring opening after reaction with ozone. Different products that form in each process may be to a varying degree less environmentally stable than the parent lamotrigine. In addition, a novel method of ozone quenching without addition of salts is presented. The new quenching method would allow subsequent mass spectrometry analysis without a solid phase extraction clean-up step. The method involves raising the pH of the sample to approximately 10 for a few seconds and lowering it back and is therefore limited to applications for which temporary pH change is not expected to affect the outcome of the analysis.

  3. Revealing the fate of cell surface human P-glycoprotein (ABCB1): The lysosomal degradation pathway.

    Science.gov (United States)

    Katayama, Kazuhiro; Kapoor, Khyati; Ohnuma, Shinobu; Patel, Atish; Swaim, William; Ambudkar, Indu S; Ambudkar, Suresh V

    2015-10-01

    P-glycoprotein (P-gp) transports a variety of chemically dissimilar amphipathic compounds including anticancer drugs. Although mechanisms of P-gp drug transport are widely studied, the pathways involving its internalization are poorly understood. The present study is aimed at elucidating the pathways involved in degradation of cell surface P-gp. The fate of P-gp at the cell surface was determined by biotinylating cell surface proteins followed by flow cytometry and Western blotting. Our data shows that the half-life of endogenously expressed P-gp is 26.7±1.1 h in human colorectal cancer HCT-15 cells. Treatment of cells with Bafilomycin A1 (BafA1) a vacuolar H+ ATPase inhibitor increased the half-life of P-gp at the cell surface to 36.1±0.5 h. Interestingly, treatment with the proteasomal inhibitors MG132, MG115 or lactacystin alone did not alter the half-life of the protein. When cells were treated with both lysosomal and proteasomal inhibitors (BafA1 and MG132), the half-life was further prolonged to 39-50 h. Functional assays done with rhodamine 123 or calcein-AM, fluorescent substrates of P-gp, indicated that the transport function of P-gp was not affected by either biotinylation or treatment with BafA1 or proteasomal inhibitors. Immunofluorescence studies done with the antibody against lysosomal marker LAMP1 and the P-gp-specific antibody UIC2 in permeabilized cells indicated that intracellular P-gp is primarily localized in the lysosomal compartment. Our results suggest that the lysosomal degradation system could be targeted to increase the sensitivity of P-gp- expressing cancer cells towards chemotherapeutic drugs.

  4. Revealing the fate of cell surface human P-glycoprotein (ABCB1): The Lysosomal Degradation Pathway

    Science.gov (United States)

    Katayama, Kazuhiro; Kapoor, Khyati; Ohnuma, Shinobu; Patel, Atish; Swaim, William; Ambudkar, Indu S.; Ambudkar, Suresh V.

    2015-01-01

    P-glycoprotein (P-gp) transports a variety of chemically dissimilar amphipathic compounds including anticancer drugs. Although mechanisms of P-gp drug transport are widely studied, the pathways involving its internalization are poorly understood. The present study is aimed at elucidating the pathways involved in degradation of cell surface P-gp. The fate of P-gp at the cell surface was determined by biotinylating cell surface proteins followed by flow cytometry and Western blotting. Our data shows that the half-life of endogenously expressed P-gp is 26.7 ± 1.1 h in human colorectal cancer HCT-15 cells. Treatment of cells with Bafilomycin A1 (BafA1) a vacuolar H+ ATPase inhibitor increased the half-life of P-gp at the cell surface to 36.1± 0.5 h. Interestingly, treatment with the proteasomal inhibitors MG132, MG115 or lactacystin alone did not alter the half-life of the protein. When cells were treated with both lysosomal and proteasomal inhibitors (BafA1 and MG132), the half-life was further prolonged to 39-50 h. Functional assays done with rhodamine 123 or calcein-AM, fluorescent substrates of P-gp, indicated that the transport function of P-gp was not affected by either biotinylation or treatment with BafA1 or proteasomal inhibitors. Immunofluorescence studies done with the antibody against lysosomal marker LAMP1 and the P-gp-specific antibody UIC2 in permeabilized cells indicated that intracellular P-gp is primarily localized in the lysosomal compartment. Our results suggest that the lysosomal degradation system could be targeted to increase the sensitivity of P-gp expressing cancer cells towards chemotherapeutic drugs. PMID:26057472

  5. Involvement of PML nuclear bodies in CBP degradation through the ubiquitin-proteasome pathway.

    Science.gov (United States)

    St-Germain, Jonathan R; Chen, Jihong; Li, Qiao

    2008-11-01

    Transcriptional coactivator CBP is involved in the regulation of an array of biological processes including cellular differentiation, proliferation and survival. The function of CBP is critical for proper embryonic development and is relevant in cancer biology. Although much is known about the functional roles of CBP in these cellular processes, fewer studies have assessed what in turn regulates CBP activity per se. It has been reported that CBP colocalizes with PML bodies which are nuclear structures disrupted in acute promyelocytic leukemia. However, the biological relevance of CBP localization to PML nuclear bodies is still unclear. In this study, we demonstrate that histone deacetylase inhibitors such as valproic acid, a therapeutically relevant compound used for the treatment of epilepsy, modulates CBP activity. Valproic acid reduces the steady-state level of CBP by inducing CBP degradation through the ubiquitin-proteasome pathway, while increasing the colocalization of CBP with ubiquitin nuclear speckles and with PML nuclear bodies. Our results suggest that PML nuclear bodies are nuclear sites involved in the ubiquitin-dependent degradation of CBP, providing novel insights in the regulation of CBP function and highlighting the relevance of its localization to PML nuclear bodies.

  6. Pathways for degrading TNT by Thu-Z: a Pantoea sp. strain.

    Science.gov (United States)

    Zou, Liangdong; Lu, Diannan; Liu, Zheng

    2012-12-01

    2,4,6-Trinitrotoluene (TNT), an extensively used and versatile explosive, is harmful in soil and water. In the present study, four bacterial strains capable of degrading TNT have been isolated from contaminated sites and named as Thu-A, Thu-B, Thu-C, and Thu-Z. Thu-Z, which gave the highest degradation efficiency compared to the others, was assigned to the genus Pantoea according to its 16S rRNA gene. Similarities in both biochemical properties and morphology suggested that Thu-Z was a Pantoea sp. strain. Thu-Z was proved to be capable of using TNT as a sole nitrogen source by cleaving NO(2) from the nitroaromatic ring by direct aromatic ring reduction. Under nitrogen-limited conditions, 96.6 % N of TNT was consumed by Thu-Z for growth, which was determined in terms of NaNO(2). Trace nitro reduction metabolites such as 2,4-diamino-6-nitrotoluene (24Dam) and 2,6-diamino-4-nitrotoluene (26Dam) were identified in the presence of (NH(4))(2)SO(4). On the other hand, 4,4',6,6'-tetranitro-2,2'-azoxytoluene (22Azo) and 2,2',6,6'-tetranitro-4,4'-azoxytoluene (44Azo) were detected in the absence of (NH(4))(2)SO(4). These indicated the existence of a dual pathway for Thu-Z, while the direct aromatic ring reduction was predominant. Addition of a nitrogen source ((NH(4))(2)SO(4)) after inoculation stimulated the growth of Thu-Z and accelerated TNT degradation.

  7. Characterization of aerobic polycyclic aromatic hydrocarbon‐degrading bacteria from Bizerte lagoon sediments, Tunisia

    National Research Council Canada - National Science Library

    Ben Said, O; Goñi‐Urriza, M.S; El Bour, M; Dellali, M; Aissa, P; Duran, R

    2008-01-01

    Aims:  To characterize polycyclic aromatic hydrocarbon (PAH)‐degrading bacteria from sediments of the Bizerte lagoon, and to determine their ability to resist other pollutants such as antibiotics and heavy metals...

  8. Molecular Characterization of 2-Chlorobiphenyl Degrading Stenotrophomonas maltophilia GS-103

    Digital Repository Service at National Institute of Oceanography (India)

    Somaraja, P.K.; Gayathri, D.; Ramaiah, N.

    and sediments. Appl Environ Microbiol 64:3014–3022 Williams WA, Lobos JH, Cheetham WE (1997) A phylogenetic analysis of aerobic polychlorinated biphenyl-degrading bacteria. Int J Syst Bacteriol 47:207–210 Xu L, Xu JJ, Jia Y, Liu WB, Jian X (2010) Congener...

  9. Characterization of nitrite degradation by Lactobacillus casei subsp. rhamnosus LCR 6013.

    Directory of Open Access Journals (Sweden)

    Dong-mei Liu

    Full Text Available Nitrites are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. The nitrite degradation capacity of Lactobacillus casei subsp. rhamnosus LCR 6013 was investigated in pickle fermentation. After LCR 6013 fermentation for 120 h at 37°C, the nitrite concentration in the fermentation system was significantly lower than that in the control sample without the LCR 6013 strain. The effects of NaCl and Vc on nitrite degradation by LCR 6013 in the De Man, Rogosa and Sharpe (MRS medium were also investigated. The highest nitrite degradations, 9.29 mg/L and 9.89 mg/L, were observed when NaCl and Vc concentrations were 0.75% and 0.02%, respectively in the MRS medium, which was significantly higher than the control group (p ≤ 0.01. Electron capture/gas chromatography and indophenol blue staining were used to study the nitrite degradation pathway of LCR 6013. The nitrite degradation products contained N2O, but no NH4(+. The LCR 6013 strain completely degraded all NaNO2 (50.00 mg/L after 16 h of fermentation. The enzyme activity of NiR in the periplasmic space was 2.5 times of that in the cytoplasm. Our results demonstrated that L. casei subsp. rhamnosus LCR 6013 can effectively degrade nitrites in both the pickle fermentation system and in MRS medium by NiR. Nitrites are degraded by the LCR 6013 strain, likely via the nitrate respiration pathway (NO2(->NO->N2O->N2, rather than the aammonium formation pathway (dissimilatory nitrate reduction to ammonium, DNRA, because the degradation products contain N2O, but not NH4(+.

  10. Characterization of purine catabolic pathway genes in coelacanths.

    Science.gov (United States)

    Forconi, Mariko; Biscotti, Maria Assunta; Barucca, Marco; Buonocore, Francesco; De Moro, Gianluca; Fausto, Anna Maria; Gerdol, Marco; Pallavicini, Alberto; Scapigliati, Giuseppe; Schartl, Manfred; Olmo, Ettore; Canapa, Adriana

    2014-09-01

    Coelacanths are a critically valuable species to explore the gene changes that took place in the transition from aquatic to terrestrial life. One interesting and biologically relevant feature of the genus Latimeria is ureotelism. However not all urea is excreted from the body; in fact high concentrations are retained in plasma and seem to be involved in osmoregulation. The purine catabolic pathway, which leads to urea production in Latimeria, has progressively lost some steps, reflecting an enzyme loss during diversification of terrestrial species. We report the results of analyses of the liver and testis transcriptomes of the Indonesian coelacanth Latimeria menadoensis and of the genome of Latimeria chalumnae, which has recently been fully sequenced in the framework of the coelacanth genome project. We describe five genes, uricase, 5-hydroxyisourate hydrolase, parahox neighbor B, allantoinase, and allantoicase, each coding for one of the five enzymes involved in urate degradation to urea, and report the identification of a putative second form of 5-hydroxyisourate hydrolase that is characteristic of the genus Latimeria. The present data also highlight the activity of the complete purine pathway in the coelacanth liver and suggest its involvement in the maintenance of high plasma urea concentrations.

  11. Isolation and characterization of a profenofos degrading bacterium

    Institute of Scientific and Technical Information of China (English)

    Saadatullah Malghani; Nivedita Chatterjee; Hu Xue Yu; Lou Zejiao

    2009-01-01

    Profenofos,a well known organophosphate pesticide,has been in agricultural use over the last two decades for controlling Lepidopteron pests of cotton and tobacco crops.In this study,a bacterial strain,OW,was isolated from a long term profenofos exposed soil by an enrichment technique and its ability to degrade profenofos was determined using gas chromatography.The isolated strain OW was identified as Pseudomonas aeruginosa according to its physiological and biochemical properties,and the analysis of its 16S rRNA gene sequence.The strain grew well at pH 5.5-7.2 with a broad temperature profile.Bioremediation of profenofos-contaminated soil was examined using soil treated with 200 μg/g profenofos,which resulted in a higher degradation rate than control soils without inoculation.In a mineral salt medium (FTW),removal in the level of profenofos of 86.81% was obtained within 48 h of incubation.The intermediates of profenofos metabolism indicated that the degradation occurred through a hydrolysis mechanism,and one of the metabolites was found to be 4 bromo-2-cholorophenol (BCP) which in turn was also mineralized by the strain.The results of this study highlighted the potentiality of P.aeruginosa as a biodegrader which could be used for the bioremediation of profenofos contaminated soil.

  12. Isolation and characterization of endosulfan-degrading bacteria from contaminated agriculture soils

    Directory of Open Access Journals (Sweden)

    Mehdi Hassanshahian

    2016-04-01

    Full Text Available Objective: To isolate and characterize endosulfan-degrading bacteria from Kerman pistachio orchards. Methods: Endosulfan-degrading bacteria were enriched in Bushnell-Hass medium. Identification and sequencing of prevalent degrading strains was performed by using PCR based on amplifying 16S rDNA. Results: The results showed that the soils of pistachio orchards have some degrading bacteria that are suitable for elimination of endosulfan from soils and the environment. Four endosulfandegrading bacteria strains belong to Achromobacter xylosoxidans (strain EN3, Pseudomonas azotoformans (strain EN4, Pseudomonas brassicacearum (strain EN7 and Pseudomonas thivervalensis (strain EN8, respectively. The best degrading strain (EN7, up to 100 mg/L, illustrated a good growth, whereas the growth was reduced in concentration higher than 100 mg/L. The results of gas chromatography confirmed the decomposition of organic pesticide by degrading-bacteria. Conclusions: By using these strains and other biological reclamation methods we can eliminate bio-environmental problems.

  13. The Whole Genome Sequence of Sphingobium chlorophenolicum L-1: Insights into the Evolution of the Pentachlorophenol Degradation Pathway

    Energy Technology Data Exchange (ETDEWEB)

    Copley, Shelley D. [University of Colorado; Rokicki, Joseph [University of Colorado; Turner, Pernilla [University of Colorado; Daligault, Hajnalka E. [Los Alamos National Laboratory (LANL); Nolan, Matt [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL

    2012-01-01

    Sphingobium chlorophenolicum Strain L-1 can mineralize the toxic pesticide pentachlorophenol (PCP). We have sequenced the genome of S. chlorophenolicum Strain L-1. The genome consists of a primary chromosome that encodes most of the genes for core processes, a secondary chromosome that encodes primarily genes that appear to be involved in environmental adaptation, and a small plasmid. The genes responsible for degradation of PCP are found on chromosome 2. We have compared the genomes of S. chlorophenolicum Strain L-1 and Sphingobium japonicum, a closely related Sphingomonad that degrades lindane. Our analysis suggests that the genes encoding the first three enzymes in the PCP degradation pathway were acquired via two different horizontal gene transfer events, and the genes encoding the final two enzymes in the pathway were acquired from the most recent common ancestor of these two bacteria.

  14. EGF Uptake and Degradation Assay to Determine the Effect of HTLV Regulatory Proteins on the ESCRT-Dependent MVB Pathway.

    Science.gov (United States)

    Murphy, Colin; Sheehy, Noreen

    2017-01-01

    The endosomal sorting complex required for transport (ESCRT) pathway plays key roles in multivesicular bodies (MVBs) formation and lysosomal degradation of membrane receptors, viral budding, and midbody abscission during cytokinesis. The epidermal growth factor receptor (EGFR) is regarded as a prototypical cargo of the MVB/ESCRT pathway and following stimulation by epidermal growth factor (EGF) EGFR/EGF complexes are internalized, sorted into MVBs, and degraded by lysosomes or recycled back to the cell membrane. Here, we describe an assay to analyze the effect of human T-cell leukemia (HTLV) regulatory proteins on the functionality of ESCRT-dependent MVB/lysosomal trafficking of EGFR/EGF complexes. This is performed by direct visualization and quantification of the rate of EGF-Alexa595/EGFR internalization and degradation in HeLa cells expressing HTLV regulatory proteins by immunofluorescence and western blot.

  15. Ubiquitin proteasome-dependent degradation of the transcriptional coactivator PGC-1{alpha} via the N-terminal pathway.

    Science.gov (United States)

    Trausch-Azar, Julie; Leone, Teresa C; Kelly, Daniel P; Schwartz, Alan L

    2010-12-17

    PGC-1α is a potent, inducible transcriptional coactivator that exerts control on mitochondrial biogenesis and multiple cellular energy metabolic pathways. PGC-1α levels are controlled in a highly dynamic manner reflecting regulation at both transcriptional and post-transcriptional levels. Here, we demonstrate that PGC-1α is rapidly degraded in the nucleus (t(½ 0.3 h) via the ubiquitin proteasome system. An N-terminal deletion mutant of 182 residues, PGC182, as well as a lysine-less mutant form, are nuclear and rapidly degraded (t(½) 0.5 h), consistent with degradation via the N terminus-dependent ubiquitin subpathway. Both PGC-1α and PGC182 degradation rates are increased in cells under low serum conditions. However, a naturally occurring N-terminal splice variant of 270 residues, NT-PGC-1α is cytoplasmic and stable (t(½>7 h), providing additional evidence that PGC-1α is degraded in the nucleus. These results strongly suggest that the nuclear N terminus-dependent ubiquitin proteasome pathway governs PGC-1α cellular degradation. In contrast, the cellular localization of NT-PCG-1α results in a longer-half-life and possible distinct temporal and potentially biological actions.

  16. Characterization of the molecular degradation mechanism of diphenyl ethers by Cupriavidus sp. WS.

    Science.gov (United States)

    Wang, Sheng; Bai, Naling; Wang, Bing; Feng, Zhuo; Hutchins, William C; Yang, Ching-Hong; Zhao, Yuhua

    2015-11-01

    Commonly used flame retardants, such as polybrominated diphenyl ethers, are extremely persistent in the environment, causing serious environmental risks. Certain strains of bacteria are able to degrade several low brominated congeners of PBDEs aerobically. However, the aerobic degradation pathway is not yet well understood, particularly at the genetic level. In this study, we isolated Cupriavidus sp. WS from the environment that could degrade diphenyl ether (DE), 4-bromodiphenyl ether, and 4,4'-bromodiphenyl ether. DE was completely degraded in 6 days without any detectable end-product. Using transposon mutagenesis, several DE degradation-deficient mutants were obtained. Knocking out bphA1, bphA2, and bphA3 eliminated the ability of the Cupriavidus sp. WS bacterium to degrade DE, indicating that the bph genes play a crucial role in DE degradation by this strain. The specific roles of bphA, bphB, and bphC were identified by systematically expressing these genes in Escherichia coli. The dihydrodiol product of BphA was dehydrogenated into 2,3-dihydroxydiphenyl ether by BphB. 2,3-Dihydroxydiphenyl ether was then decomposed into phenol and 2-pyrone-6-carboxylic acid by BphC. Thus, BphA, BphB, and BphC act sequentially in the aerobic degradation of DE, 4-bromodiphenyl ether, and 4,4'-dibromodiphenyl ether by the Cupriavidus sp. WS bacterium.

  17. Insights from 14C into C loss pathways in degraded peatlands

    Science.gov (United States)

    Evans, Martin; Evans, Chris; Allott, Tim; Stimson, Andrew; Goulsbra, Claire

    2016-04-01

    Peatlands are important global stores of terrestrial carbon. Lowered water tables due to changing climate and direct or indirect human intervention produce a deeper aerobic zone and have the potential to enhance loss of stored carbon from the peat profile. The quasi continuous accumulation of organic matter in active peatlands means that the age of fluvial dissolved organic carbon exported from peatland systems is related to the source depth in the peat profile. Consequently 14C analysis of DOC in waters draining peatlands has the potential not only to tell us about the source of fluvial carbon and the stability of the peatland but also about the dominant hydrological pathways in the peatland system. This paper will present new radiocarbon determinations from peatland streams draining the heavily eroded peatlands of the southern Pennine uplands in the UK. These blanket peatland systems are highly degraded, with extensive bare peat and gully erosion resulting from air pollution during the industrial revolution, overgrazing, wildfire and climatic changes. Deep and extensive gullying has significantly modified the hydrology of these systems leading to local and more widespread drawdown of water table. 14C data from DOC in drainage waters are presented from two catchments; one with extensive gully erosion and the other with a combination of gully erosion and sheet erosion of the peat. At the gully eroded site DOC in drainage waters is as old as 160 BP but at the site with extensive sheet erosion dates of up to 1069 BP are amongst the oldest recorded from blanket peatland globally These data indicate significant degradation of stored carbon from the eroding peatlands. Initial comparisons of the 14C data with modelled water table for the catchments and depth-age curves for catchment peats suggests that erosion of the peat surface, allowing decomposition of exposed older organic material is a potential mechanism producing aged carbon from the eroded catchment. This

  18. Preparation and characterization of a degradable magnesium phosphate bone cement.

    Science.gov (United States)

    Yu, Ying; Xu, Chao; Dai, Honglian

    2016-12-01

    A kind of degradable magnesium phosphate bone cement (MPBC) was fabricated by using the mixed powders of magnesium oxide (MgO), potassium dihydrogen phosphate (KH2PO4) and calcium dihydrogen phosphate (Ca(H2PO4)2.H2O). As MgKPO4, the main product of MgO and KH2PO4 was alkaline, the Ca(H2PO4)2.H2O was added to neutralize the alkali of the system. And the effects of Ca(H2PO4)2.H2O on the performance of MPBC were discussed. The results showed that the adding of Ca(H2PO4)2.H2O extended the setting time, which was about 6 min to 18 min. The compressive strength increased first and then decreased, and maximum value reached 31.2 MPa after setting for 24 h without any additional pressure. The MPBC was degradable in Tris-HCl solution, and the extracts of the cytotoxicity assay showed that the MPBC had good biocompatibility, indicating that the MPBC had good biodegradable and biocompatible properties.

  19. ISOLATION AND CHARACTERIZATION OF CRUDE OIL DEGRADING BACILLUS SPP.

    Directory of Open Access Journals (Sweden)

    A. Akhavan Sepahi, I. Dejban Golpasha, M. Emami, A. M. Nakhoda

    2008-07-01

    Full Text Available Today, application of microorganisms for removing crude oil pollution from contaminated sites as bioremediation studies, was considered by scientists because other methods such as surfactant washing and incineration lead to production of more toxic compounds and they are non-economic. Fifteen crude oil degrading bacillus spp. were isolated from contaminated sites. Two isolated showed best growth in liquid media with 1-3% (v/v crude oil and mineral salt medium, then studied for enzymatic activities on tested media. The results showed maximal increase in optical densities and total viable count concomitant with decrease in pH on fifth day of experimental period for bacillus S6. Typical generation time on mineral salt with 1% crude oil is varying between 18-20h, 25-26h respectively for bacillus S6 and S35. Total protein was monitored at determined time intervals as biodegradation indices. Increasing of protein concentration during the incubation period reveals that isolated bacillus can degrade crude oil and increase microbial biomass. These bacillus spp. reduced surface tension from 60 (mN/m to 31 and 38 (mN/m, It means that these bacillus spp. can produce sufficient surfactant and have good potential of emulsification capacity. The results demonstrated that these bacillus spp. can utilize crude oil as a carbon and energy source.

  20. Dysregulation of protein degradation pathways may mediate the liver injury and phospholipidosis associated with a cationic amphiphilic antibiotic drug

    Energy Technology Data Exchange (ETDEWEB)

    Mosedale, Merrie [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); Wu, Hong [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Kurtz, C. Lisa [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); Schmidt, Stephen P. [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Adkins, Karissa, E-mail: Karissa.Adkins@pfizer.com [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Harrill, Alison H. [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); University of Arkansas for Medical Sciences, Little Rock, AR72205 (United States)

    2014-10-01

    A large number of antibiotics are known to cause drug-induced liver injury in the clinic; however, interpreting clinical risk is not straightforward owing to a lack of predictivity of the toxicity by standard preclinical species and a poor understanding of the mechanisms of toxicity. An example is PF-04287881, a novel ketolide antibiotic that caused elevations in liver function tests in Phase I clinical studies. In this study, a mouse diversity panel (MDP), comprised of 34 genetically diverse, inbred mouse strains, was utilized to model the toxicity observed with PF-04287881 treatment and investigate potential mechanisms that may mediate the liver response. Significant elevations in serum alanine aminotransferase (ALT) levels in PF-04287881-treated animals relative to vehicle-treated controls were observed in the majority (88%) of strains tested following a seven day exposure. The average fold elevation in ALT varied by genetic background and correlated with microscopic findings of hepatocellular hypertrophy, hepatocellular single cell necrosis, and Kupffer cell vacuolation (confirmed as phospholipidosis) in the liver. Global liver mRNA expression was evaluated in a subset of four strains to identify transcript and pathway differences that distinguish susceptible mice from resistant mice in the context of PF-04287881 treatment. The protein ubiquitination pathway was highly enriched among genes associated with PF-04287881-induced hepatocellular necrosis. Expression changes associated with PF-04287881-induced phospholipidosis included genes involved in drug transport, phospholipid metabolism, and lysosomal function. The findings suggest that perturbations in genes involved in protein degradation leading to accumulation of oxidized proteins may mediate the liver injury induced by this drug. - Highlights: • Identified susceptible and resistant mouse strains to liver injury induced by a CAD • Liver injury characterized by single cell necrosis, and phospholipidosis

  1. Cytolethal distending toxins require components of the ER-associated degradation pathway for host cell entry.

    Directory of Open Access Journals (Sweden)

    Aria Eshraghi

    2014-07-01

    Full Text Available Intracellular acting protein exotoxins produced by bacteria and plants are important molecular determinants that drive numerous human diseases. A subset of these toxins, the cytolethal distending toxins (CDTs, are encoded by several Gram-negative pathogens and have been proposed to enhance virulence by allowing evasion of the immune system. CDTs are trafficked in a retrograde manner from the cell surface through the Golgi apparatus and into the endoplasmic reticulum (ER before ultimately reaching the host cell nucleus. However, the mechanism by which CDTs exit the ER is not known. Here we show that three central components of the host ER associated degradation (ERAD machinery, Derlin-2 (Derl2, the E3 ubiquitin-protein ligase Hrd1, and the AAA ATPase p97, are required for intoxication by some CDTs. Complementation of Derl2-deficient cells with Derl2:Derl1 chimeras identified two previously uncharacterized functional domains in Derl2, the N-terminal 88 amino acids and the second ER-luminal loop, as required for intoxication by the CDT encoded by Haemophilus ducreyi (Hd-CDT. In contrast, two motifs required for Derlin-dependent retrotranslocation of ERAD substrates, a conserved WR motif and an SHP box that mediates interaction with the AAA ATPase p97, were found to be dispensable for Hd-CDT intoxication. Interestingly, this previously undescribed mechanism is shared with the plant toxin ricin. These data reveal a requirement for multiple components of the ERAD pathway for CDT intoxication and provide insight into a Derl2-dependent pathway exploited by retrograde trafficking toxins.

  2. Cytolethal distending toxins require components of the ER-associated degradation pathway for host cell entry.

    Science.gov (United States)

    Eshraghi, Aria; Dixon, Shandee D; Tamilselvam, Batcha; Kim, Emily Jin-Kyung; Gargi, Amandeep; Kulik, Julia C; Damoiseaux, Robert; Blanke, Steven R; Bradley, Kenneth A

    2014-07-01

    Intracellular acting protein exotoxins produced by bacteria and plants are important molecular determinants that drive numerous human diseases. A subset of these toxins, the cytolethal distending toxins (CDTs), are encoded by several Gram-negative pathogens and have been proposed to enhance virulence by allowing evasion of the immune system. CDTs are trafficked in a retrograde manner from the cell surface through the Golgi apparatus and into the endoplasmic reticulum (ER) before ultimately reaching the host cell nucleus. However, the mechanism by which CDTs exit the ER is not known. Here we show that three central components of the host ER associated degradation (ERAD) machinery, Derlin-2 (Derl2), the E3 ubiquitin-protein ligase Hrd1, and the AAA ATPase p97, are required for intoxication by some CDTs. Complementation of Derl2-deficient cells with Derl2:Derl1 chimeras identified two previously uncharacterized functional domains in Derl2, the N-terminal 88 amino acids and the second ER-luminal loop, as required for intoxication by the CDT encoded by Haemophilus ducreyi (Hd-CDT). In contrast, two motifs required for Derlin-dependent retrotranslocation of ERAD substrates, a conserved WR motif and an SHP box that mediates interaction with the AAA ATPase p97, were found to be dispensable for Hd-CDT intoxication. Interestingly, this previously undescribed mechanism is shared with the plant toxin ricin. These data reveal a requirement for multiple components of the ERAD pathway for CDT intoxication and provide insight into a Derl2-dependent pathway exploited by retrograde trafficking toxins.

  3. Ribosomal Protein Mutations Result in Constitutive p53 Protein Degradation through Impairment of the AKT Pathway.

    Directory of Open Access Journals (Sweden)

    Ana T Antunes

    2015-07-01

    Full Text Available Mutations in ribosomal protein (RP genes can result in the loss of erythrocyte progenitor cells and cause severe anemia. This is seen in patients with Diamond-Blackfan anemia (DBA, a pure red cell aplasia and bone marrow failure syndrome that is almost exclusively linked to RP gene haploinsufficiency. While the mechanisms underlying the cytopenia phenotype of patients with these mutations are not completely understood, it is believed that stabilization of the p53 tumor suppressor protein may induce apoptosis in the progenitor cells. In stark contrast, tumor cells from zebrafish with RP gene haploinsufficiency are unable to stabilize p53 even when exposed to acute DNA damage despite transcribing wild type p53 normally. In this work we demonstrate that p53 has a limited role in eliciting the anemia phenotype of zebrafish models of DBA. In fact, we find that RP-deficient embryos exhibit the same normal p53 transcription, absence of p53 protein, and impaired p53 response to DNA damage as RP haploinsufficient tumor cells. Recently we reported that RP mutations suppress activity of the AKT pathway, and we show here that this suppression results in proteasomal degradation of p53. By re-activating the AKT pathway or by inhibiting GSK-3, a downstream modifier that normally represses AKT signaling, we are able to restore the stabilization of p53. Our work indicates that the anemia phenotype of zebrafish models of DBA is dependent on factors other than p53, and may hold clinical significance for both DBA and the increasing number of cancers revealing spontaneous mutations in RP genes.

  4. Ozonation degradation of microcystin-LR in aqueous solution: intermediates, byproducts and pathways.

    Science.gov (United States)

    Chang, Jing; Chen, Zhong-lin; Wang, Zhe; Shen, Ji-min; Chen, Qian; Kang, Jing; Yang, Lei; Liu, Xiao-wei; Nie, Chang-xin

    2014-10-15

    The intermediates and byproducts formed during the ozonation of microcystin-LR (MC-LR, m/z = 995.5) and the probable degradation pathway were investigated at different initial molar ratios of ozone to MC-LR ([O3]0/[MC-LR]0). Seven reaction intermediates with m/z ≥ 795.4 were observed by LC/MS, and four of them (m/z = 815.4, 827.3, 853.3 and 855.3) have not been previously reported. Meanwhile, six aldehyde-based byproducts with molecular weights of 30-160 were detected for the first time. Intermediates structures demonstrated that ozone reacted with two sites of MC-LR: the diene bonds in the Adda side chain and the Mdha amino acid in the cyclic structure. The fragment from the Adda side chain oxidative cleavage could be further oxidized to an aldehyde with a molecular weight of 160 at low [O3]0/[MC-LR]0. Meanwhile, the polypeptide structure of MC-LR was difficult to be further oxidized, unless [O3]0/[MC-LR]0 > 10. After further oxidation of the intermediates, five other aldehyde-based byproducts were detected by GC/MS: formaldehyde, acetaldehyde, isovaleraldehyde, glyoxal and methylglyoxal. Formaldehyde, isovaleraldehyde and methylglyoxal were the dominant species. The yields of the aldehydes varied greatly, depending on the value of [O3]0/[MC-LR]0.

  5. REGγ regulates ERα degradation via ubiquitin–proteasome pathway in breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Chai, Fan; Liang, Yan [Breast Disease Center, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China); Bi, Jiong [Laboratory of General Surgery, First Affiliated Hospital, Sun Yet-sen University, Guangzhou 510080 (China); Chen, Li; Zhang, Fan [Breast Disease Center, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China); Cui, Youhong [Institute of Pathology and Southwest Cancer Center, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China); Jiang, Jun, E-mail: jcbd@medmail.com.cn [Breast Disease Center, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China)

    2015-01-02

    Highlights: • High expression of REGγ is correlated with ERα status and poor clinical features. • Cell growth, mobility and invasion are significantly impaired by REGγ knockdown. • REGγ indirectly regulates ERα protein expression. - Abstract: REGγ is a proteasome coactivator which regulates proteolytic activity in eukaryotic cells. Abundant lines of evidence have showed that REGγ is over expressed in a number of human carcinomas. However, its precise role in the pathogenesis of cancer is still unclear. In this study, by examining 200 human breast cancer specimens, we demonstrated that REGγ was highly expressed in breast cancers, and the expression of REGγ was positively correlated with breast cancer patient estrogen receptor alpha (ERα) status. Moreover, the expression of REGγ was found positively associated with poor clinical features and low survival rates in ERα positive breast cancer patients. Further cell culture studies using MCF7 and BT474 breast cancer cell lines showed that cell proliferation, motility, and invasion capacities were decreased significantly by REGγ knockdown. Lastly, we demonstrated that REGγ indirectly regulates the degradation of ERα protein via ubiquitin–proteasome pathway. In conclusion, our findings provide the evidence that REGγ expression was positively correlated with ERα status and poor clinical prognosis in ERα positive breast cancer patients. As well, we disclose a new connection between the two molecules that are both highly expressed in most breast cancer cases.

  6. Functional characterization of Sporothrix schenckii glycosidases involved in the N-linked glycosylation pathway.

    Science.gov (United States)

    Lopes-Bezerra, Leila M; Lozoya-Pérez, Nancy E; López-Ramírez, Luz A; Martínez-Álvarez, José A; Teixeira, Marcus M; Felipe, Maria S S; Flores-Carreón, Arturo; Mora-Montes, Héctor M

    2015-01-01

    Protein glycosylation pathways are conserved metabolic processes in eukaryotic organisms and are required for cell fitness. In fungal pathogens, the N-linked glycosylation pathway is indispensable for proper cell wall composition and virulence. In Sporothrix schenckii sensu stricto, the causative agent of sporotrichosis, little is known about this glycosylation pathway. Here, using a genome-wide screening for putative members of the glycosyl hydrolase (CAZy - GH) families 47 and 63, which group enzymes involved in the processing step during N-linked glycan maturation, we found seven homologue genes belonging to family 47 and one to family 63. The eight genes were individually expressed in C. albicans null mutants lacking either MNS1 (for members of family 47) or CWH41 (for the member of family 63). Our results indicate that SsCWH41 is the functional ortholog of CaCWH41, whereas SsMNS1 is the functional ortholog of CaMNS1. The remaining genes of family 47 encode Golgi mannosidases and endoplasmic reticulum degradation-enhancing alpha-mannosidase-like proteins (EDEMs). Since these GH families gather proteins used as target for drugs to control cell growth, identification of these genes could help in the design of antifungals that could be used to treat sporotrichosis and other fungal diseases. In addition, to our knowledge, we are the first to report that Golgi mannosidases and EDEMs are expressed and characterized in yeast cells.

  7. Characterization of atrial natriuretic peptide degradation by cell-surface peptidase activity on endothelial cells

    Science.gov (United States)

    Frost, S. J.; Whitson, P. A.

    1993-01-01

    Atrial natriuretic peptide (ANP) is a fluid-regulating peptide hormone that promotes vasorelaxation, natriuresis, and diuresis. The mechanisms for the release of ANP and for its clearance from the circulation play important roles in modulating its biological effects. Recently, we have reported that the cell surface of an endothelial cell line, CPA47, could degrade 125I-ANP in the presence of EDTA. In this study, we have characterized this degradation of 125I-ANP. The kinetics of ANP degradation by the surface of CPA47 cells were first order, with a Km of 320 +/- 60 nM and Vmax of 35 +/- 14 pmol of ANP degraded/10 min/10(5) cells at pH 7.4. ANP is degraded by the surface of CPA47 cells over a broad pH range from 7.0-8.5. Potato carboxypeptidase inhibitor and bestatin inhibited 125I-ANP degradation, suggesting that this degradative activity on the surface of CPA47 cells has exopeptidase characteristics. The selectivity of CPA47 cell-surface degradation of ANP was demonstrated when 125I-ANP degradation was inhibited in the presence of neuropeptide Y and angiotensin I and II but not bradykinin, bombesin, endothelin-1, or substance P. The C-terminal amino acids phe26 and tyr28 were deduced to be important for ANP interaction with the cell-surface peptidase(s) based on comparison of the IC50 of various ANP analogues and other natriuretic peptides for the inhibition of ANP degradation. These data suggest that a newly characterized divalent cation-independent exopeptidase(s) that selectively recognizes ANP and some other vasoactive peptides exists on the surface of endothelial cells.

  8. Blu-Ray-based micromechanical characterization platform for biopolymer degradation assessment

    DEFF Research Database (Denmark)

    Casci Ceccacci, Andrea; Chen, Ching-Hsiu; Hwu, En-Te

    2017-01-01

    Degradable biopolymers are used as carrier materials in drug delivery devices. A complete understanding of their degradation behaviour is thus crucial in the design of new delivery systems. Here we combine a reliable method, based on spray coated micromechanical resonators and a disposable...... microfluidic chip, to characterize biopolymer degradation under the action of enzymes in controlled flow condition. The sensing platform is based on the mechanics and optics from a Blu-Ray player, which automatically localize individual sensors within the array, and sequentially measure and record...

  9. Isolation and characterization of organic-sulfur degradation bacterial strain

    Institute of Scientific and Technical Information of China (English)

    YANG Yu; DIAO Meng-xue; SHI Wu-yang; LI Li; DAI Qin-yun; QIU Guan-zhou

    2007-01-01

    A bacterial strain that was capable of degrading organic sulfur (dibenzothiophene) was isolated by enrichment techniques from the petroleum-contaminated soil collected from Zhongyuan Oil Field. The strain is named ZYX and is gram-positive.This strain undergoes bacilus-coccus morphological change, and forms yellow-pigment glossy circular colonies with 1.5 mm in diameter on average after 2 d incubation on Luria-Bertani(LB) plates. The full-length of 16S rDNA sequence of strain ZYX was determined and analyzed. Strain ZYX is found most relative with the genus of Arthrobacter. The similarity values between ZYX and Arthrobacter sp. P2 is 99.53%. The main morphological, biochemical and physiological features of strain ZYX accord with those of Arthrobacter. It is found that the optimal initial pH for growth is about 7.0, and the optimal concentration of dibenzothiophene(DBT)for growth is 0.10 g/L. Additionally, the results show that the best carbon source and nitrogen source are glycerol and glutamine,respectively.

  10. An approach of habitat degradation assessment for characterization on coastal habitat conservation tendency.

    Science.gov (United States)

    Zhou, Xi-Yin; Lei, Kun; Meng, Wei

    2017-09-01

    Coastal zones are population and economy highly intensity regions all over the world, and coastal habitat supports the sustainable development of human society. The accurate assessment of coastal habitat degradation is the essential prerequisite for coastal zone protection. In this study, an integrated framework of coastal habitat degradation assessment including landuse classification, habitat classifying and zoning, evaluation criterion of coastal habitat degradation and coastal habitat degradation index has been established for better regional coastal habitat assessment. Through establishment of detailed three-class landuse classification, the fine landscape change is revealed, the evaluation criterion of coastal habitat degradation through internal comparison based on the results of habitat classifying and zoning could indicate the levels of habitat degradation and distinguish the intensity of human disturbances in different habitat subareas under the same habitat classification. Finally, the results of coastal habitat degradation assessment could be achieved through coastal habitat degradation index (CHI). A case study of the framework is carried out in the Circum-Bohai-Sea-Coast, China, and the main results show the following: (1) The accuracy of all land use classes are above 90%, which indicates a satisfactory accuracy for the classification map. (2) The Circum-Bohai-Sea-Coast is divided into 3 kinds of habitats and 5 subareas. (3) In the five subareas of the Circum-Bohai-Sea-Coast, the levels of coastal habitat degradation own significant difference. The whole Circum-Bohai-Sea-Coast generally is in a worse state according to area weighting of each habitat subarea. This assessment framework of coastal habitat degradation would characterize the landuse change trend, realize better coastal habitat degradation assessment, reveal the habitat conservation tendency and distinguish intensity of human disturbances. Furthermore, it would support for accurate coastal

  11. Characterization of return flow pathways during flood irrigation

    Science.gov (United States)

    Claes, N.; Paige, G. B.; Parsekian, A.; Gordon, B. L.; Miller, S. N.

    2015-12-01

    With a decline in water resources available for private consumption and irrigation, the importance of sustainable water management practices is increasing. Local management decisions, based on models may affect the availability of water both locally and downstream, causing a ripple effect. It is therefore important that the models that these local management decisions are based on, accurately quantify local hydrological processes and the timescales at which they happen. We are focusing on return flow from flood irrigation, which can occur via different pathways back to the streams: overland flow, near-surface return flow and return flow via pathways below the vadose zone. The question addressed is how these different pathways each contribute to the total amount of return flow and the dynamics behind them. We used time-lapse ERT measurements in combination with an ensemble of ERT and seismic lines to answer this question via (1) capturing the process of gradual fragmentation of aqueous environments in the vadose zone during drying stages at field scale; (2) characterization of the formation of preferential flow paths from infiltrating wetting fronts during wetting cycles at field scale. The time-lapse ERT provides the possibility to capture the dynamic processes involved during the occurrence of finger flow or macro-pores when an intensive wetting period during flood irrigation occurs. It elucidates the dynamics of retention in the vadose zone during drying and wetting periods at field scale. This method provides thereby a link to upscale from laboratory experiments to field scale and watershed scale for finger flow and preferential flow paths and illustrates the hysteresis behavior at field scale.

  12. Multi-parametric MRI characterization of enzymatically degraded articular cartilage.

    Science.gov (United States)

    Nissi, Mikko J; Salo, Elli-Noora; Tiitu, Virpi; Liimatainen, Timo; Michaeli, Shalom; Mangia, Silvia; Ellermann, Jutta; Nieminen, Miika T

    2016-07-01

    Several laboratory and rotating frame quantitative MRI parameters were evaluated and compared for detection of changes in articular cartilage following selective enzymatic digestion. Bovine osteochondral specimens were subjected to 44 h incubation in control medium or in collagenase or chondroitinase ABC to induce superficial collagen or proteoglycan (glycosaminoglycan) alterations. The samples were scanned at 9.4 T for T1 , T1 Gd (dGEMRIC), T2 , adiabatic T1 ρ , adiabatic T2 ρ , continuous-wave T1 ρ , TRAFF2 , and T1 sat relaxation times and for magnetization transfer ratio (MTR). For reference, glycosaminoglycan content, collagen fibril orientation and biomechanical properties were determined. Changes primarily in the superficial cartilage were noted after enzymatic degradation. Most of the studied parameters were sensitive to the destruction of collagen network, whereas glycosaminoglycan depletion was detected only by native T1 and T1 Gd relaxation time constants throughout the tissue and by MTR superficially. T1 , adiabatic T1 ρ , adiabatic T2 ρ , continuous-wave T1 ρ , and T1 sat correlated significantly with the biomechanical properties while T1 Gd correlated with glycosaminoglycan staining. The findings indicated that most of the studied MRI parameters were sensitive to both glycosaminoglycan content and collagen network integrity, with changes due to enzymatic treatment detected primarily in the superficial tissue. Strong correlation of T1 , adiabatic T1ρ , adiabatic T2 ρ , continuous-wave T1 ρ , and T1 sat with the altered biomechanical properties, reflects that these parameters were sensitive to critical functional properties of cartilage. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1111-1120, 2016.

  13. Isolation and characterization of RDX-degrading Rhodococcus species from a contaminated aquifer.

    Science.gov (United States)

    Bernstein, Anat; Adar, Eilon; Nejidat, Ali; Ronen, Zeev

    2011-09-01

    Groundwater contamination by the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a global problem. Israel's coastal aquifer was contaminated with RDX. This aquifer is mostly aerobic and we therefore sought aerobic bacteria that might be involved in natural attenuation of the compound in the aquifer. RDX-degrading bacteria were captured by passively sampling the indigenous bacteria onto sterile sediments placed within sampling boreholes. Aerobic RDX biodegradation potential was detected in the sediments sampled from different locations along the plume. RDX degradation with the native sampled consortium was accompanied by 4-nitro-2,4-diazabutanal formation. Two bacterial strains of the genus Rhodococcus were isolated from the sediments and identified as aerobic RDX degraders. The xplA gene encoding the cytochrome P450 enzyme was partially (~500 bp) sequenced from both isolates. The obtained DNA sequences had 99% identity with corresponding gene fragments of previously isolated RDX-degrading Rhodococcus strains. RDX degradation by both strains was prevented by 200 μM of the cytochrome P450 inhibitor metyrapone, suggesting that cytochrome P450 indeed mediates the initial step in RDX degradation. RDX biodegradation activity by the T7 isolate was inhibited in the presence of nitrate or ammonium concentrations above 1.6 and 5.5 mM, respectively (100 mg l(-1)) while the T9N isolate's activity was retarded only by ammonium concentrations above 5.5 mM. This study shows that bacteria from the genus Rhodococcus, potentially degrade RDX in the saturated zone as well, following the same aerobic degradation pathway defined for other Rhodococcus species. RDX-degrading activity by the Rhodococcus species isolate T9N may have important implications for the bioremediation of nitrate-rich RDX-contaminated aquifers.

  14. A vacuolar carboxypeptidase mutant of Arabidopsis thaliana is degraded by the ERAD pathway independently of its N-glycan

    Energy Technology Data Exchange (ETDEWEB)

    Yamamoto, Masaya; Kawanabe, Mitsuyoshi; Hayashi, Yoko; Endo, Toshiya [Department of Chemistry, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602 (Japan); Nishikawa, Shuh-ichi, E-mail: shuh@biochem.chem.nagoya-u.ac.jp [Department of Chemistry, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602 (Japan)

    2010-03-12

    Misfolded proteins produced in the endoplasmic reticulum (ER) are degraded by a mechanism, the ER-associated degradation (ERAD). Here we report establishment of the experimental system to analyze the ERAD in plant cells. Carboxypeptidase Y (CPY) is a vacuolar enzyme and its mutant CPY* is degraded by the ERAD in yeast. Since Arabidopsis thaliana has AtCPY, an ortholog of yeast CPY, we constructed and expressed fusion proteins consisting of AtCPY and GFP and of AtCPY*, which carries a mutation homologous to yeast CPY*, and GFP in A. thaliana cells. While AtCPY-GFP was efficiently transported to the vacuole, AtCPY*-GFP was retained in the ER to be degraded in proteasome- and Cdc48-dependent manners. We also found that AtCPY*-GFP was degraded by the ERAD in yeast cells, but that its single N-glycan did not function as a degradation signal in yeast or plant cells. Therefore, AtCPY*-GFP can be used as a marker protein to analyze the ERAD pathway, likely for nonglycosylated substrates, in plant cells.

  15. Electrochemical treatment of iopromide under conditions of reverse osmosis concentrates--elucidation of the degradation pathway.

    Science.gov (United States)

    Lütke Eversloh, C; Henning, N; Schulz, M; Ternes, T A

    2014-01-01

    Application of reverse osmosis for the reuse of treated wastewater on the one hand offers a way to provide high quality effluent waters. On the other hand reverse osmosis concentrates exhibiting highly concentrated contaminants are produced simultaneously. Electrochemical treatment of those concentrates is regarded as one possible answer to the problem of their disposal into surface waters. Nevertheless, due to the diversity of direct and indirect degradation processes during electrolysis, special care has to be taken about the formation of toxic transformation products (TPs). In this study the electrochemical transformation of the X-ray contrast medium iopromide was investigated as a representative of biologically persistent compounds. For this purpose, anodic oxidation at boron doped diamond as well as cathodic reduction using a platinum electrode were considered. Kinetic analyses revealed a transformation of 100 μM iopromide with first order kinetic constants between 0.6 and 1.6 × 10(-4) s(-1) at the beginning and a subsequent increase of the reaction order due to the influence of secondary oxidants formed during electrolysis. Mineralization up to 96% was achieved after about 7.5 h. At shorter treatment times several oxidatively and reductively formed transformation products were detected, whereas deiodinated iopromide represented the major fraction. Nevertheless, the latter exhibited negligible toxicological relevance according to tests on vibrio fisheri. Additional experiments utilizing a divided cell setup enabled the elucidation of the transformation pathway, whereas emerging TPs could be identified by means of high resolution mass spectrometry and MS(n)-fragmentations. During electrolysis the iodine released from Iopromide was found to 90% as iodide and to 10% as iodate even in the open cell experiments, limiting the potential formation of toxic iodo-disinfection by-products. Chlorinated TPs were not found.

  16. Assessing degradation and recovery pathways in lakes impacted by eutrophication using the sediment record

    Directory of Open Access Journals (Sweden)

    Helen eBennion

    2015-08-01

    Full Text Available Efforts to restore enriched lakes have increased yet there remains uncertainty about whether restoration targets can be achieved and over what timescale. Paleoecological techniques, principally diatom analyses, were used to examine the degree of impact and recovery in 12 European lakes subject to eutrophication and subsequent reduction in nutrient loading. Dissimilarity scores showed that all sites experienced progressive deviation from the reference sample (core bottom prior to nutrient reduction, and principal curves indicated gradual compositional change with enrichment. When additive models were applied to the latter, the changes were statistically significant in 9 of the 12 sites. Shifts in diatom composition following reduction in nutrient loading were more equivocal, with a reversal towards the reference flora seen only in four of the deep lakes and one of the shallow lakes. Of these, only two were significant (Lake Bled and Mjøsa. Alternative nutrient sources seem to explain the lack of apparent recovery in the other deep lakes. In three shallow lakes diatom assemblages were replaced by a community associated with lower productivity but not the one seen prior to enrichment. Internal loading and top down control may influence recovery in shallow lakes and climate change may have confounded recovery in several of the study sites. Hence, ecosystem recovery is not simply a reversal of the degradation pathway and may take several decades to complete or, for some lakes, may not take place at all. By assessing ecological change over a decadal to centennial timescale, the study highlights the important role that paleolimnology can play in establishing a benchmark against which managers can evaluate the degree to which their restoration efforts are successful.

  17. MIR125B1 represses the degradation of the PML-RARA oncoprotein by an autophagy-lysosomal pathway in acute promyelocytic leukemia.

    Science.gov (United States)

    Zeng, Cheng-Wu; Chen, Zhen-Hua; Zhang, Xing-Ju; Han, Bo-Wei; Lin, Kang-Yu; Li, Xiao-Juan; Wei, Pan-Pan; Zhang, Hua; Li, Yangqiu; Chen, Yue-Qin

    2014-10-01

    Acute promyelocytic leukemia (APL) is characterized by the t(15;17)-associated PML-RARA fusion gene. We have previously found that MIR125B1 is highly expressed in patients with APL and may be associated with disease pathogenesis; however, the mechanism by which MIR125B1 exerts its oncogenic potential has not been fully elucidated. Here, we demonstrated that MIR125B1 abundance correlates with the PML-RARA status. MIR125B1 overexpression enhanced PML-RARA expression and inhibited the ATRA-induced degradation of the PML-RARA oncoprotein. RNA-seq analysis revealed a direct link between the PML-RARA degradation pathway and MIR125B1-arrested differentiation. We further demonstrated that the MIR125B1-mediated blockade of PML-RARA proteolysis was regulated via an autophagy-lysosomal pathway, contributing to the inhibition of APL differentiation. Furthermore, we identified DRAM2 (DNA-damage regulated autophagy modulator 2), a critical regulator of autophagy, as a novel target that was at least partly responsible for the function of MIR125B1 involved in autophagy. Importantly, the knockdown phenotypes for DRAM2 are similar to the effects of overexpressing MIR125B1 as impairment of PML-RARA degradation, inhibition of autophagy, and myeloid cell differentiation arrest. These effects of MIR125B1 and its target DRAM2 were further confirmed in an APL mouse model. Thus, MIR125B1 dysregulation may interfere with the effectiveness of ATRA-mediated differentiation through an autophagy-dependent pathway, representing a novel potential APL therapeutic target.

  18. Synchrotron-Based in Situ Characterization of the Scaffold Mass Loss from Erosion Degradation

    Directory of Open Access Journals (Sweden)

    Nahshon K. Bawolin

    2016-07-01

    Full Text Available The mass loss behavior of degradable tissue scaffolds is critical to their lifespan and other degradation-related properties including mechanical strength and mass transport characteristics. This paper presents a novel method based on synchrotron imaging to characterize the scaffold mass loss from erosion degradation in situ, or without the need of extracting scaffolds once implanted. Specifically, the surface-eroding degradation of scaffolds in a degrading medium was monitored in situ by synchrotron-based imaging; and the time-dependent geometry of scaffolds captured by images was then employed to estimate their mass loss with time, based on the mathematical model that was adopted from the literature of surface erosion with the experimentally-identified model parameters. Acceptable agreement between experimental results and model predictions was observed for scaffolds in a cylindrical shape, made from poly(lactic-co-glycolic acid (PLGA and polycaprolactone (PCL. This study illustrates that geometry evaluation by synchrotron-based imaging is an effective means to in situ characterize the scaffold mass loss as well as possibly other degradation-related properties.

  19. Electro-catalytic oxidation of phenacetin with a three-dimensional reactor: Degradation pathway and removal mechanism.

    Science.gov (United States)

    Xiao, Mengshi; Zhang, Yonggang

    2016-06-01

    Phenacetin is a common analgesic, anti-arthritic and anti-rheumatic drug. This study dealt with the degradation of phenacetin in alkaline media using a three-dimensional reactor with particle electrodes. Particular attention was paid to the degradation pathway and the reaction mechanism in the system. Liquid chromatography coupled with time-of-flight mass spectrometry was used to identify the intermediates. The phenacetin was observed to be firstly cut off the branch chains main by direct oxidation, and then the intermediates further degraded to ring opening products by hydroxyl radical resulting from indirect oxidation and finally mineralized to CO2, H2O. A possible removal mechanism was proposed that direct and indirect oxidation together did effect on the pollutants with oxygen.

  20. Isolation and characterization of endosulfan-degrading bacteria from contaminated agriculture soils

    Institute of Scientific and Technical Information of China (English)

    Mehdi Hassanshahian; Zahra Shahi

    2016-01-01

    Objective: To isolate and characterize endosulfan-degrading bacteria from Kerman pistachio orchards. Methods: Endosulfan-degrading bacteria were enriched in Bushnell-Hass medium. Identification and sequencing of prevalent degrading strains was performed by usingPCR based on amplifying16S rDNA. Results: The results showed that the soils of pistachio orchards have some degrading bacteria that are suitable for elimination of endosulfan from soils and the environment. Four endosulfan-degrading bacteria strains belong toAchromobacter xylosoxidans (strain EN3),Pseudomonas azotoformans (strain EN4),Pseudomonas brassicacearum (strain EN7) andPseudomonas thivervalensis (strain EN8), respectively. The best degrading strain (EN7), up to 100 mg/L, illustrated a good growth, whereas the growth was reduced in concentration higher than 100 mg/L. The results of gas chromatography confirmed the decomposition of organic pesticide by degrading-bacteria. Conclusions: By using these strains and other biological reclamation methods we can eliminate bio-environmental problems.

  1. Characterizing pesticide sorption and degradation in microscale biopurification systems using column displacement experiments.

    Science.gov (United States)

    De Wilde, Tineke; Mertens, Jan; Simunek, Jirka; Sniegowksi, Kristel; Ryckeboer, Jaak; Jaeken, Peter; Springael, Dirk; Spanoghe, Pieter

    2009-02-01

    Biopurification systems treating pesticide contaminated water are very efficient, however they operate as a black box. Processes inside the system are not yet characterized. To optimize the performance, knowledge of degradation and retention processes needs to be generated. Therefore, displacement experiments were carried out for four pesticides (isoproturon, bentazone, metalaxyl, linuron) in columns containing different organic mixtures. Bromide, isoproturon and bentazone breakthrough curves (BTCs) were well described using the convection-dispersion equation (CDE) and a first-order degradation kinetic approach. Metalaxyl and linuron BTCs were well described using the CDE model expanded with Monod-type kinetics. Freundlich sorption, first-order degradation and Monod kinetics coefficients were fitted to the BTCs. Fitted values of the distribution coefficient K(f,column) were much lower than those determined from batch experiments. Based on mobility, pesticides were ranked as: bentazone>metalaxyl-isoproturon>linuron. Based on degradability, pesticides were ranked as: linuron>metalaxyl-isoproturon>bentazone.

  2. Characterization and Degradation of Masonry Mortar in Historic Brick Structures

    Directory of Open Access Journals (Sweden)

    Denis A. Brosnan

    2014-01-01

    Full Text Available This study characterized mortars from a masonry fortification in Charleston, South Carolina (USA, harbor where construction was during the period 1839–1860. This location for analysis was interesting because of the sea water impingement on the structure. The study was included as part of an overall structural assessment with restoration as an objective. The mortars were found to be cement, lime, and sand mixtures in proportions similar to ones expected from the historic literature, that is, one part binder to two parts of sand. The binder was found to be American natural cement, a substance analogous to the European Roman cement. The results suggest that the thermal history of the cement during manufacturing affected setting rate explaining why the cements were considered as variable during the mid-to-late 1800s. Fine pores were found in mortars exposed to sea water resulting from corrosion. Contemporary natural cement was shown to release calcium in aqueous solution. While this release of calcium is necessary for setting in natural and Portland cements, excessive calcium solution, as exacerbated by sea water contact and repointing with Portland cement mortars, was shown to result in brick scaling or decay through cryptoflorescence.

  3. Characterization and degradation of pectin derived from Budimka apple

    Directory of Open Access Journals (Sweden)

    MILOS V. NIKOLIC

    2008-02-01

    Full Text Available The characterization of apple pectin and its oligogalacturonic fractions derived from the autochthones apple variety Budimka, characteristic for central Serbia, is described in this paper. After extraction, the apple pectin was subjected to controlled enzymatic hydrolysis by polygalacturonase (PG and pectin lyase (PL from Aspergillus niger and then fractionated by ion-exchange column chromatography on Dowex 1X-8 (200–400 mesh. Saturated oligogalacturonic acids, obtained by controlled hydrolysis with PG, were efficiently separated by elution with a gradient of Na acetate buffer (pH 6.0, while unsaturated oligogalacturonic acids, obtained by controlled hydrolysis with PL, were separated on the same resin, using a gradient of Na formate buffer (pH 4.7 as the eluent. The yields of the fractions with the particular degree of polymerization (DP were also determined. The total content of neutral saccharides in the original Budimka apple pectin was detected by HPLC analysis of the 4-nitrobenzoyl derivatives of the sugar, and amounted to 5.31 %. Among the neutral saccharides, contents of galactose, glucose, rhamnose, arabinose, xylose and mannose were detected.

  4. Genetic associations of type 2 diabetes with islet amyloid polypeptide processing and degrading pathways in asian populations.

    Directory of Open Access Journals (Sweden)

    Vincent Kwok Lim Lam

    Full Text Available Type 2 diabetes (T2D is a complex disease characterized by beta cell dysfunctions. Islet amyloid polypeptide (IAPP is highly conserved and co-secreted with insulin with over 40% of autopsy cases of T2D showing islet amyloid formation due to IAPP aggregation. Dysregulation in IAPP processing, stabilization and degradation can cause excessive oligomerization with beta cell toxicity. Previous studies examining genetic associations of pathways implicated in IAPP metabolism have yielded conflicting results due to small sample size, insufficient interrogation of gene structure and gene-gene interactions. In this multi-staged study, we screened 89 tag single nucleotide polymorphisms (SNPs in 6 candidate genes implicated in IAPP metabolism and tested for independent and joint associations with T2D and beta cell dysfunctions. Positive signals in the stage-1 were confirmed by de novo and in silico analysis in a multi-centre unrelated case-control cohort. We examined the association of significant SNPs with quantitative traits in a subset of controls and performed bioinformatics and relevant functional analyses. Amongst the tag SNPs, rs1583645 in carboxypeptidase E (CPE and rs6583813 in insulin degrading enzyme (IDE were associated with 1.09 to 1.28 fold increased risk of T2D (P Meta = 9.4×10(-3 and 0.02 respectively in a meta-analysis of East Asians. Using genetic risk scores (GRS with each risk variant scoring 1, subjects with GRS≥3 (8.2% of the cohort had 56% higher risk of T2D than those with GRS = 0 (P = 0.01. In a subcohort of control subjects, plasma IAPP increased and beta cell function index declined with GRS (P = 0.008 and 0.03 respectively. Bioinformatics and functional analyses of CPE rs1583645 predicted regulatory elements for chromatin modification and transcription factors, suggesting differential DNA-protein interactions and gene expression. Taken together, these results support the importance of dysregulation of IAPP

  5. Genetic Associations of Type 2 Diabetes with Islet Amyloid Polypeptide Processing and Degrading Pathways in Asian Populations

    Science.gov (United States)

    Lam, Vincent Kwok Lim; Ma, Ronald Ching Wan; Lee, Heung Man; Hu, Cheng; Park, Kyong Soo; Furuta, Hiroto; Wang, Ying; Tam, Claudia Ha Ting; Sim, Xueling; Ng, Daniel Peng-Keat; Liu, Jianjun; Wong, Tien-Yin; Tai, E. Shyong; Morris, Andrew P.; Tang, Nelson Leung Sang; Woo, Jean; Leung, Ping Chung; Kong, Alice Pik Shan; Ozaki, Risa; Jia, Wei Ping; Lee, Hong Kyu; Nanjo, Kishio; Xu, Gang; Ng, Maggie Chor Yin; So, Wing-Yee; Chan, Juliana Chung Ngor

    2013-01-01

    Type 2 diabetes (T2D) is a complex disease characterized by beta cell dysfunctions. Islet amyloid polypeptide (IAPP) is highly conserved and co-secreted with insulin with over 40% of autopsy cases of T2D showing islet amyloid formation due to IAPP aggregation. Dysregulation in IAPP processing, stabilization and degradation can cause excessive oligomerization with beta cell toxicity. Previous studies examining genetic associations of pathways implicated in IAPP metabolism have yielded conflicting results due to small sample size, insufficient interrogation of gene structure and gene-gene interactions. In this multi-staged study, we screened 89 tag single nucleotide polymorphisms (SNPs) in 6 candidate genes implicated in IAPP metabolism and tested for independent and joint associations with T2D and beta cell dysfunctions. Positive signals in the stage-1 were confirmed by de novo and in silico analysis in a multi-centre unrelated case-control cohort. We examined the association of significant SNPs with quantitative traits in a subset of controls and performed bioinformatics and relevant functional analyses. Amongst the tag SNPs, rs1583645 in carboxypeptidase E (CPE) and rs6583813 in insulin degrading enzyme (IDE) were associated with 1.09 to 1.28 fold increased risk of T2D (PMeta = 9.4×10−3 and 0.02 respectively) in a meta-analysis of East Asians. Using genetic risk scores (GRS) with each risk variant scoring 1, subjects with GRS≥3 (8.2% of the cohort) had 56% higher risk of T2D than those with GRS = 0 (P = 0.01). In a subcohort of control subjects, plasma IAPP increased and beta cell function index declined with GRS (P = 0.008 and 0.03 respectively). Bioinformatics and functional analyses of CPE rs1583645 predicted regulatory elements for chromatin modification and transcription factors, suggesting differential DNA-protein interactions and gene expression. Taken together, these results support the importance of dysregulation of IAPP metabolism in

  6. New Method to Characterize Degradation of First Surface Aluminum Reflectors: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Sutter, F.; Heller, P.; Meyen, S.; Pitz-Paal, R.; Kennedy, C.; Fernandez-Garcia, A.; Schmucker, M.

    2010-10-01

    This paper reports the development of a new optical instrument capable of characterizing the aging process of enhanced first surface aluminum reflectors for concentrating solar power (CSP) application. Samples were exposed outdoors at different sites and in accelerated exposure tests. All samples exposed outdoors showed localized corrosion spots. Degradation originated from points of damage in the protective coating, but propagated underneath the protective coating. The degraded samples were analyzed with a microscope and with a newly designed space-resolved specular reflectometer (SR)2 that is capable of optically detecting and characterizing the corrosion spots. The device measures the specular reflectance at three acceptance angles and the wavelengths with spatial resolution using a digital camera's CMOS sensor. It can be used to measure the corrosion growth rate during outdoor and accelerated exposure tests. These results will allow a correlation between the degraded mirror surface and its specular reflectance.

  7. Microbial Degradation of Indole and Its Derivatives

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Arora

    2015-01-01

    Full Text Available Indole and its derivatives, including 3-methylindole and 4-chloroindole, are environmental pollutants that are present worldwide. Microbial degradation of indole and its derivatives can occur in several aerobic and anaerobic pathways; these pathways involve different known and characterized genes. In this minireview, we summarize and explain the microbial degradation of indole, indole-3-acetic acid, 4-chloroindole, and methylindole.

  8. Effect of surface chemistry of Fe-Ni nanoparticles on mechanistic pathways of azo dye degradation.

    Science.gov (United States)

    Bokare, Alok D; Chikate, Rajeev C; Rode, Chandrashekhar V; Paknikar, Kishore M

    2007-11-01

    The degradation of Orange G, a monoazo dye, in aqueous solutions was investigated using as-synthesized and stored Fe-Ni bimetallic nanoparticles. Batch experiments with a nanocatalyst loading of 3 g/L showed complete dye degradation (150 mg/L) after 10 min of reaction time. HPLC-MS analysis of the degradation products showed that as-synthesized nanoparticles reductively cleaved the azo linkage to produce aniline as the major degradation product. However, 1-year-stored nanoparticles showed an oxidative degradation of Orange G through a hydroxyl-radical induced coupling of parent and/or product molecules. XPS analysis in corroboration with HPLC-MS data showed that the surface chemistry between Fe and Ni in as-synthesized and stored nanoparticles play a crucial role in directing the mode of degradation. Reductive dye degradation using as-synthesized nanoparticles proceeded through hydride transfer from nickel, whereas formation of a Fe2+ -Ni(0) galvanic cell in stored nanoparticles generated hydroxyl radicals from water in a nonFenton type reaction. The latter were responsible for the generation of radical centers on the dye molecule, which led to a coupling-mediated oxidative degradation of Orange G. The generation of hydroxyl radicals is further substantiated with radical quenching experiments using ascorbic acid indicating that stored nanoparticles degrade Orange G through a predominantly oxidative mechanism. HPLC-MS and XPS analysis of dye degradation using as-synthesized nanoparticles exposed to air and water confirmed that the reductive or oxidative degradation capability of Fe-Ni nanoparticles is decided by the time and type of catalyst aging process.

  9. Genetic characterization of caffeine degradation by bacteria and its potential applications.

    Science.gov (United States)

    Summers, Ryan M; Mohanty, Sujit K; Gopishetty, Sridhar; Subramanian, Mani

    2015-05-01

    The ability of bacteria to grow on caffeine as sole carbon and nitrogen source has been known for over 40 years. Extensive research into this subject has revealed two distinct pathways, N-demethylation and C-8 oxidation, for bacterial caffeine degradation. However, the enzymological and genetic basis for bacterial caffeine degradation has only recently been discovered. This review article discusses the recent discoveries of the genes responsible for both N-demethylation and C-8 oxidation. All of the genes for the N-demethylation pathway, encoding enzymes in the Rieske oxygenase family, reside on 13.2-kb genomic DNA fragment found in Pseudomonas putida CBB5. A nearly identical DNA fragment, with homologous genes in similar orientation, is found in Pseudomonas sp. CES. Similarly, genes for C-8 oxidation of caffeine have been located on a 25.2-kb genomic DNA fragment of Pseudomonas sp. CBB1. The C-8 oxidation genes encode enzymes similar to those found in the uric acid metabolic pathway of Klebsiella pneumoniae. Various biotechnological applications of these genes responsible for bacterial caffeine degradation, including bio-decaffeination, remediation of caffeine-contaminated environments, production of chemical and fuels and development of diagnostic tests have also been demonstrated. © 2015 The Author. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  10. TRIM22 Inhibits the TRAF6-stimulated NF-κB Pathway by Targeting TAB2 for Degradation

    Institute of Scientific and Technical Information of China (English)

    Hui Qiu; Fang Huang; Han Xiao; Binlian Sun; Rongge Yang

    2013-01-01

    Tripartite motif containing 22 (TRIM22),a member of the TRIM/RBCC family,has been reported to activate the nuclear factor-kappa B (NF-κB) pathway in unstimulated macrophage cell lines,but the detailed mechanisms governing this activation remains unclear.We investigated this mechanism in HEK293T cells.We found that overexpression of TRIM22 could activate the NF-κB pathway and conversely,could inhibit the tumor necrosis factor receptor-associated factor 6 (TRAF6)-stimulated NF-κB pathway in HEK293T cells.Further experiments showed that TRIM22 could decrease the self-ubiquitination of TRAF6,and interact with and degrade transforming growth factor-β activated kinase 1 binding protein 2 (TAB2),and that these effects could be partially rescued by a TRIM22 RING domain deletion mutant.Collectively,our data indicate that overexpression of TRIM22 may negatively regulate the TRAF6-stimulated NF-κB pathway by interacting with and degrading TAB2.

  11. Genetic Characterization of the Galactitol Utilization Pathway of Salmonella enterica Serovar Typhimurium.

    Science.gov (United States)

    Nolle, Nicoletta; Felsl, Angela; Heermann, Ralf; Fuchs, Thilo M

    2017-02-15

    Galactitol degradation by salmonellae remains underinvestigated, although this metabolic capability contributes to growth in animals (R. R. Chaudhuri et al., PLoS Genet 9:e1003456, 2013, https://doi.org/10.1371/journal.pgen.1003456). The genes responsible for this metabolic capability are part of a 9.6-kb gene cluster that spans from gatY to gatR (STM3253 to STM3262) and encodes a phosphotransferase system, four enzymes, and a transporter of the major facilitator superfamily. Genome comparison revealed the presence of this genetic determinant in nearly all Salmonella strains. The generation time of Salmonella enterica serovar Typhimurium strain ST4/74 was higher in minimal medium with galactitol than with glucose. Knockout of STM3254 and gatC resulted in a growth-deficient phenotype of S Typhimurium, with galactitol as the sole carbon source. Partial deletion of gatR strongly reduced the lag phase of growth with galactitol, whereas strains overproducing GatR exhibited a near-zero growth phenotype. Luciferase reporter assays demonstrated strong induction of the gatY and gatZ promoters, which control all genes of this cluster except gatR, in the presence of galactitol but not glucose. Purified GatR bound to these two main gat gene cluster promoters as well as to its own promoter, demonstrating that this autoregulated repressor controls galactitol degradation. Surface plasmon resonance spectroscopy revealed distinct binding properties of GatR toward the three promoters, resulting in a model of differential gat gene expression. The cyclic AMP receptor protein (CRP) bound these promoters with similarly high affinities, and a mutant lacking crp showed severe growth attenuation, demonstrating that galactitol utilization is subject to catabolite repression. Here, we provide the first genetic characterization of galactitol degradation in Salmonella, revealing novel insights into the regulation of this dissimilatory pathway.

  12. Water deficit induces chlorophyll degradation via the 'PAO/phyllobilin' pathway in leaves of homoio- (Craterostigma pumilum) and poikilochlorophyllous (Xerophyta viscosa) resurrection plants.

    Science.gov (United States)

    Christ, Bastien; Egert, Aurélie; Süssenbacher, Iris; Kräutler, Bernhard; Bartels, Dorothea; Peters, Shaun; Hörtensteiner, Stefan

    2014-11-01

    Angiosperm resurrection plants exhibit poikilo- or homoiochlorophylly as a response to water deficit. Both strategies are generally considered as effective mechanisms to reduce oxidative stress associated with photosynthetic activity under water deficiency. The mechanism of water deficit-induced chlorophyll (Chl) degradation in resurrection plants is unknown but has previously been suggested to occur as a result of non-enzymatic photooxidation. We investigated Chl degradation during dehydration in both poikilochlorophyllous (Xerophyta viscosa) and homoiochlorophyllous (Craterostigma pumilum) species. We demonstrate an increase in the abundance of PHEOPHORBIDE a OXYGENASE (PAO), a key enzyme of Chl breakdown, together with an accumulation of phyllobilins, that is, products of PAO-dependent Chl breakdown, in both species. Phyllobilins and PAO levels diminished again in leaves from rehydrated plants. We conclude that water deficit-induced poikilochlorophylly occurs via the well-characterized PAO/phyllobilin pathway of Chl breakdown and that this mechanism also appears conserved in a resurrection species displaying homoiochlorophylly. The roles of the PAO/phyllobilin pathway during different plant developmental processes that involve Chl breakdown, such as leaf senescence and desiccation, fruit ripening and seed maturation, are discussed. © 2014 John Wiley & Sons Ltd.

  13. A calmodulin-like protein suppresses RNA silencing and promotes geminivirus infection by degrading SGS3 via the autophagy pathway in Nicotiana benthamiana

    Science.gov (United States)

    Li, Fangfang; Zhao, Nan; Xu, Xiongbiao; Wang, Yaqin; Yang, Xiuling; Liu, Shu-Sheng; Wang, Aiming; Zhou, Xueping

    2017-01-01

    A recently characterized calmodulin-like protein is an endogenous RNA silencing suppressor that suppresses sense-RNA induced post-transcriptional gene silencing (S-PTGS) and enhances virus infection, but the mechanism underlying calmodulin-like protein-mediated S-PTGS suppression is obscure. Here, we show that a calmodulin-like protein from Nicotiana benthamiana (NbCaM) interacts with Suppressor of Gene Silencing 3 (NbSGS3). Deletion analyses showed that domains essential for the interaction between NbSGS3 and NbCaM are also required for the subcellular localization of NbSGS3 and NbCaM suppressor activity. Overexpression of NbCaM reduced the number of NbSGS3-associated granules by degrading NbSGS3 protein accumulation in the cytoplasm. This NbCaM-mediated NbSGS3 degradation was sensitive to the autophagy inhibitors 3-methyladenine and E64d, and was compromised when key autophagy genes of the phosphatidylinositol 3-kinase (PI3K) complex were knocked down. Meanwhile, silencing of key autophagy genes within the PI3K complex inhibited geminivirus infection. Taken together these data suggest that NbCaM acts as a suppressor of RNA silencing by degrading NbSGS3 through the autophagy pathway. PMID:28212430

  14. Hydrocarbon Degradation Pathways used by Coastal Sediment Microbial Communities exposed to Crude Oil

    Science.gov (United States)

    Spaulding-Astudillo, F.; Sharrar, A.; Orcutt, B.

    2016-02-01

    The site-specific microbial community response to crude oil exposure in marine environments is not well described. Moreover, the abundance of genes implicated in long-chain alkane (LCA) and polycyclic aromatic hydrocarbon (PAH) degradation are not well understood. Coastal sediments from the Beaufort Sea, Gulf of Alaska, and Portland Harbor were treated with crude oil and incubated aerobically. Deep-sea sediments from the Gulf of Mexico were treated with the same crude oil and anaerobically incubated in situ for five months before recovery. Cycloclasticus, a known hydrocarbon-degrader, was abundant in all oiled, aerobic samples regardless of temperature, demonstrating a generalist oil-response strategy. Other hydrocarbon degrading bacteria showed differential response to either site or temperature. Primers for alkB, assA, bssA, and ncr, catabolic gene markers for aerobic LCA degradation, anaerobic LCA degradation, anaerobic LCA & PAH degradation, and anaerobic PAH degradation, respectively, were found in literature and tested on DNA extracts in a QPCR-based assay. Gene abundance was site and condition variable.

  15. Metabolic analysis of the soil microbe Dechloromonas aromatica str. RCB: indications of a surprisingly complex life-style and cryptic anaerobic pathways for aromatic degradation

    Directory of Open Access Journals (Sweden)

    Feil Helene

    2009-08-01

    Full Text Available Abstract Background Initial interest in Dechloromonas aromatica strain RCB arose from its ability to anaerobically degrade benzene. It is also able to reduce perchlorate and oxidize chlorobenzoate, toluene, and xylene, creating interest in using this organism for bioremediation. Little physiological data has been published for this microbe. It is considered to be a free-living organism. Results The a priori prediction that the D. aromatica genome would contain previously characterized "central" enzymes to support anaerobic aromatic degradation of benzene proved to be false, suggesting the presence of novel anaerobic aromatic degradation pathways in this species. These missing pathways include the benzylsuccinate synthase (bssABC genes (responsible for fumarate addition to toluene and the central benzoyl-CoA pathway for monoaromatics. In depth analyses using existing TIGRfam, COG, and InterPro models, and the creation of de novo HMM models, indicate a highly complex lifestyle with a large number of environmental sensors and signaling pathways, including a relatively large number of GGDEF domain signal receptors and multiple quorum sensors. A number of proteins indicate interactions with an as yet unknown host, as indicated by the presence of predicted cell host remodeling enzymes, effector enzymes, hemolysin-like proteins, adhesins, NO reductase, and both type III and type VI secretory complexes. Evidence of biofilm formation including a proposed exopolysaccharide complex and exosortase (epsH are also present. Annotation described in this paper also reveals evidence for several metabolic pathways that have yet to be observed experimentally, including a sulphur oxidation (soxFCDYZAXB gene cluster, Calvin cycle enzymes, and proteins involved in nitrogen fixation in other species (including RubisCo, ribulose-phosphate 3-epimerase, and nif gene families, respectively. Conclusion Analysis of the D. aromatica genome indicates there is much to be

  16. Metabolic analysis of the soil microbe Dechloromonas aromatica str. RCB: indications of a surprisingly complex life-style and cryptic anaerobic pathways for aromatic degradation

    Energy Technology Data Exchange (ETDEWEB)

    Salinero, Kennan Kellaris; Keller, Keith; Feil, William S.; Feil, Helene; Trong, Stephan; Di Bartolo, Genevieve; Lapidus, Alla

    2008-11-17

    Initial interest in Dechloromonas aromatica strain RCB arose from its ability to anaerobically degrade benzene. It is also able to reduce perchlorate and oxidize chlorobenzoate, toluene, and xylene, creating interest in using this organism for bioremediation. Little physiological data has been published for this microbe. It is considered to be a free-living organism. The a priori prediction that the D. aromatica genome would contain previously characterized 'central' enzymes involved in anaerobic aromatic degradation proved to be false, suggesting the presence of novel anaerobic aromatic degradation pathways in this species. These missing pathways include the benzyl succinyl synthase (bssABC) genes (responsible for formate addition to toluene) and the central benzoylCoA pathway for monoaromatics. In depth analyses using existing TIGRfam, COG, and InterPro models, and the creation of de novo HMM models, indicate a highly complex lifestyle with a large number of environmental sensors and signaling pathways, including a relatively large number of GGDEF domain signal receptors and multiple quorum sensors. A number of proteins indicate interactions with an as yet unknown host, as indicated by the presence of predicted cell host remodeling enzymes, effector enzymes, hemolysin-like proteins, adhesins, NO reductase, and both type III and type VI secretory complexes. Evidence of biofilm formation including a proposed exopolysaccharide complex with the somewhat rare exosortase (epsH), is also present. Annotation described in this paper also reveals evidence for several metabolic pathways that have yet to be observed experimentally, including a sulphur oxidation (soxFCDYZAXB) gene cluster, Calvin cycle enzymes, and nitrogen fixation (including RubisCo, ribulose-phosphate 3-epimerase, and nif gene families, respectively). Analysis of the D. aromatica genome indicates there is much to be learned regarding the metabolic capabilities, and life-style, for this microbial

  17. Multi-technique approach for qualitative and quantitative characterization of furazidin degradation kinetics under alkaline conditions.

    Science.gov (United States)

    Bērziņš, Kārlis; Kons, Artis; Grante, Ilze; Dzabijeva, Diana; Nakurte, Ilva; Actiņš, Andris

    2016-09-10

    Degradation of drug furazidin was studied under different conditions of environmental pH (11-13) and temperature (30-60°C). The novel approach of hybrid hard- and soft-multivariate curve resolution-alternating least squares (HS-MCR-ALS) method was applied to UV-vis spectral data to determine a valid kinetic model and kinetic parameters of the degradation process. The system was found to be comprised of three main species and best characterized by two consecutive first-order reactions. Furazidin degradation rate was found to be highly dependent on the applied environmental conditions, showing more prominent differences between both degradation steps towards higher pH and temperature. Complimentary qualitative analysis of the degradation process was carried out using HPLC-DAD-TOF-MS. Based on the obtained chromatographic and mass spectrometric results, as well as additional computational analysis of the species (theoretical UV-vis spectra calculations utilizing TD-DFT methodology), the operating degradation mechanism was proposed to include formation of a 5-hydroxyfuran derivative, followed by complete hydrolysis of furazidin hydantoin ring.

  18. Characterizing the Degradation of Alginate Hydrogel for Use in Multilumen Scaffolds for Spinal Cord Repair.

    Science.gov (United States)

    Shahriari, Dena; Koffler, Jacob; Lynam, Daniel A; Tuszynski, Mark H; Sakamoto, Jeffrey S

    2015-10-21

    Alginate was studied as a degradable nerve guidance scaffold material in vitro and in vivo. In vitro degradation rates were determined using rheology to measure the change in shear modulus vs time. The shear modulus decreased from 155 kPa to 5 kPa within 2 days; however, alginate samples maintained their superficial geometry for over 28 days. The degradation behavior was supported by materials characterization data showing alginate consisted of high internal surface area (400 m(2) /g), which likely facilitated the release of cross-linking cations resulting in the rapid decrease in shear modulus. To assess the degradation rate in vivo, multilumen scaffolds were fabricated using a fiber templating technique. The scaffolds were implanted in a 2 mm-long T3 full transection rodent spinal cord lesion model for 14 days. Although there was some evidence of axon guidance, in general, alginate scaffolds degraded before axons could grow over the 2 mm-long lesion. Enabling alginate-based scaffolds for nerve repair will likely require approaches to slow its degradation. This article is protected by copyright. All rights reserved.

  19. Glutamine supplementation stimulates protein-synthetic and inhibits protein-degradative signaling pathways in skeletal muscle of diabetic rats.

    Directory of Open Access Journals (Sweden)

    Adriana C Lambertucci

    Full Text Available In this study, we investigated the effect of glutamine (Gln supplementation on the signaling pathways regulating protein synthesis and protein degradation in the skeletal muscle of rats with streptozotocin (STZ-induced diabetes. The expression levels of key regulatory proteins in the synthetic pathways (Akt, mTOR, GSK3 and 4E-BP1 and the degradation pathways (MuRF-1 and MAFbx were determined using real-time PCR and Western blotting in four groups of male Wistar rats; 1 control, non-supplemented with glutamine; 2 control, supplemented with glutamine; 3 diabetic, non-supplemented with glutamine; and 4 diabetic, supplemented with glutamine. Diabetes was induced by the intravenous injection of 65 mg/kg bw STZ in citrate buffer (pH 4.2; the non-diabetic controls received only citrate buffer. After 48 hours, diabetes was confirmed in the STZ-treated animals by the determination of blood glucose levels above 200 mg/dL. Starting on that day, a solution of 1 g/kg bw Gln in phosphate buffered saline (PBS was administered daily via gavage for 15 days to groups 2 and 4. Groups 1 and 3 received only PBS for the same duration. The rats were euthanized, and the soleus muscles were removed and homogenized in extraction buffer for the subsequent measurement of protein and mRNA levels. The results demonstrated a significant decrease in the muscle Gln content in the diabetic rats, and this level increased toward the control value in the diabetic rats receiving Gln. In addition, the diabetic rats exhibited a reduced mRNA expression of regulatory proteins in the protein synthesis pathway and increased expression of those associated with protein degradation. A reduction in the skeletal muscle mass in the diabetic rats was observed and was alleviated partially with Gln supplementation. The data suggest that glutamine supplementation is potentially useful for slowing the progression of muscle atrophy in patients with diabetes.

  20. Vitreous State Characterization of Pharmaceutical Compounds Degrading upon Melting by Using Fast Scanning Calorimetry.

    Science.gov (United States)

    Corvis, Yohann; Wurm, Andreas; Schick, Christoph; Espeau, Philippe

    2015-06-04

    Fast scanning calorimetry, a technique mainly devoted to polymer characterization, is applied here for the first time to low molecular mass organic compounds that degrade upon melting, such as ascorbic acid and prednisolone. Due to the fast scan rates upon heating and cooling, the substances can be obtained in the molten state without degradation and then quenched into the glassy state. The hydrated form and the polymorphic Form 1 of prednisolone were investigated. It is shown that once the sesquihydrate dehydrates, a molten product is obtained. Depending on the heating rate, this molten phase may recrystallize or not into Form 1.

  1. Identification of dimethylamine monooxygenase in marine bacteria reveals a metabolic bottleneck in the methylated amine degradation pathway.

    Science.gov (United States)

    Lidbury, Ian; Mausz, Michaela A; Scanlan, David J; Chen, Yin

    2017-07-01

    Methylated amines (MAs) are ubiquitous in the marine environment and their subsequent flux into the atmosphere can result in the formation of aerosols and ultimately cloud condensation nuclei. Therefore, these compounds have a potentially important role in climate regulation. Using Ruegeria pomeroyi as a model, we identified the genes encoding dimethylamine (DMA) monooxygenase (dmmABC) and demonstrate that this enzyme degrades DMA to monomethylamine (MMA). Although only dmmABC are required for enzyme activity in recombinant Escherichia coli, we found that an additional gene, dmmD, was required for the growth of R. pomeroyi on MAs. The dmmDABC genes are absent from the genomes of multiple marine bacteria, including all representatives of the cosmopolitan SAR11 clade. Consequently, the abundance of dmmDABC in marine metagenomes was substantially lower than the genes required for other metabolic steps of the MA degradation pathway. Thus, there is a genetic and potential metabolic bottleneck in the marine MA degradation pathway. Our data provide an explanation for the observation that DMA-derived secondary organic aerosols (SOAs) are among the most abundant SOAs detected in fine marine particles over the North and Tropical Atlantic Ocean.

  2. A novel role for ATM in regulating proteasome-mediated protein degradation through suppression of the ISG15 conjugation pathway.

    Directory of Open Access Journals (Sweden)

    Laurence M Wood

    Full Text Available Ataxia Telangiectasia (A-T is an inherited immunodeficiency disorder wherein mutation of the ATM kinase is responsible for the A-T pathogenesis. Although the precise role of ATM in A-T pathogenesis is still unclear, its function in responding to DNA damage has been well established. Here we demonstrate that in addition to its role in DNA repair, ATM also regulates proteasome-mediated protein turnover through suppression of the ISG15 pathway. This conclusion is based on three major pieces of evidence: First, we demonstrate that proteasome-mediated protein degradation is impaired in A-T cells. Second, we show that the reduced protein turnover is causally linked to the elevated expression of the ubiquitin-like protein ISG15 in A-T cells. Third, we show that expression of the ISG15 is elevated in A-T cells derived from various A-T patients, as well as in brain tissues derived from the ATM knockout mice and A-T patients, suggesting that ATM negatively regulates the ISG15 pathway. Our current findings suggest for the first time that proteasome-mediated protein degradation is impaired in A-T cells due to elevated expression of the ISG15 conjugation pathway, which could contribute to progressive neurodegeneration in A-T patients.

  3. Mechanism and Reaction Pathways for Microcystin-LR Degradation through UV/H2O2 Treatment.

    Science.gov (United States)

    Liu, Yafeng; Ren, Jing; Wang, Xiangrong; Fan, Zhengqiu

    2016-01-01

    Microcystin-LR (MCLR) is the most common cyanotoxin in contaminated aquatic systems. MCLR inhibits protein phosphatases 1 and 2A, leading to liver damage and tumor formation. MCLR is relatively stable owing to its cyclic structures. The combined UV/H2O2 technology can degrade MCLR efficiently. The second-order rate constant of the reaction between MCLR and hydroxyl radical (·OH) is 2.79(±0.23)×1010 M-1 s-1 based on the competition kinetics model using nitrobenzene as reference compound. The probable degradation pathway was analyzed through liquid chromatography mass spectrometry. Results suggested that the major destruction pathways of MCLR were initiated by ·OH attack on the benzene ring and diene of the Adda side chain. The corresponding aldehyde or ketone peptide residues were formed through further oxidation. Another minor destruction pathway involved ·OH attack on the methoxy group of the Adda side chain, followed by complete removal of the methoxy group. The combined UV/H2O2 system is a promising technology for MCLR removal in contaminated aquatic systems.

  4. Isolation and characterization of diesel degrading bacteria, Sphingomonas sp. and Acinetobacter junii from petroleum contaminated soil

    Science.gov (United States)

    Zhang, Qiuzhuo; Wang, Duanchao; Li, Mengmeng; Xiang, Wei-Ning; Achal, Varenyam

    2014-03-01

    Two indigenous bacteria of petroleum contaminated soil were characterized to utilize diesel fuel as the sole carbon and energy sources in this work. 16S rRNA gene sequence analysis identified these bacteria as Sphingomonas sp. and Acinetobacter junii. The ability to degrade diesel fuel has been demonstrated for the first time by these isolates. The results of IR analyses showed that Sphingomonas sp. VA1 and A. junii VA2 degraded up to 82.6% and 75.8% of applied diesel over 15 days, respectively. In addition, Sphingomonas sp. VA1 possessed the higher cellular hydrophobicities of 94% for diesel compared to 81% by A. junii VA2. The isolates Sphingomonas sp. VA1 and A. junii VA2 exhibited 24% and 18%, respectively emulsification activity. This study reports two new diesel degrading bacterial species, which can be effectively used for bioremediation of petroleum contaminated sites.

  5. Impedance Characterization of the Degradation of Insulating Layer Patterned on Interdigitated Microelectrode.

    Science.gov (United States)

    Lee, Gihyun; Kim, Sohee; Cho, Sungbo

    2015-10-01

    Life-time and functionality of planar microelectrode-based devices are determined by not only the corrosion-resistance of the electrode, but also the durability of the insulation layer coated on the transmission lines. Degradation of the insulating layer exposed to a humid environment or solution may cause leakage current or signal loss, and a decrease in measurement sensitivity. In this study, degradation of SU-8, an epoxy-based negative photoresist and insulating material, patterned on Au interdigitated microelectrode (IDE) for long-term (>30 days) immersion in an electrolyte at 37 °C was investigated by electrical impedance spectroscopy and theoretical equivalent circuit modeling. From the experiment and simulation results, it was found that the degradation level of the insulating layer of the IDE electrode can be characterized by monitoring the resistance of the insulating layer among the circuit parameters of the designed equivalent circuit modeling.

  6. Photochemical degradation of atrazine in UV and UV/H2O2 process: pathways and toxic effects of products.

    Science.gov (United States)

    Choi, Hyun-Jin; Kim, Daekeun; Lee, Tae-Jin

    2013-01-01

    The degradation of atrazine in aqueous solution by UV or UV/H2O2 processes, and the toxic effects of the degradation products were explored. The mineralization of atrazine was not observed in the UV irradiation process, resulting in the production of hydroxyatrazine (OIET) as the final product. In the UV/H2O2 process, the final product was ammeline (OAAT), which was obtained by two different pathways of reaction: dechlorination followed by hydroxylation, and the de-alkylation of atrazine. The by-products of the reaction of dechlorination followed by hydroxylation were OIET and hydroxydeethyl atrazine (OIAT), and those of de-alkylation were deisopropyl atrazine (CEAT), deethyl atrazine (CIAT), and deethyldeisopropyl atrazine (CAAT). OIAT and OAAT appeared to be quite stable in the degradation of atrazine by the UV/H2O2 process. In a toxicity test using Daphnia magna, the acute toxic unit (TUa) was less than 1 of TUa (100/EC50, %) in the UV/H2O2 process after 30 min of reaction time, while 1.2 to 1.3 of TUa was observed in the UV process. The TUa values of atrazine and the degradation products have the following decreasing order: OIET> Atrazine> CEAT≈CIAT> CAAT. OIAT and OAAT did not show any toxic effects.

  7. Photolytic and photocatalytic degradation of quinclorac in ultrapure and paddy field water: identification of transformation products and pathways.

    Science.gov (United States)

    Pareja, Lucía; Pérez-Parada, Andrés; Agüera, Ana; Cesio, Verónica; Heinzen, Horacio; Fernández-Alba, Amadeo R

    2012-05-01

    Quinclorac (QNC) is an effective but rather persistent herbicide commonly used in rice production. This herbicide presents a mean persistence in the environment so its residues are considered of environmental relevance. However, few studies have been conducted to investigate its environmental behavior and degradation. In the present work, direct photolysis and TiO(2) photocatalysis of the target compound in ultrapure and paddy field water were investigated. After 10h photolysis in ultrapure water, the concentration of QNC declined 26% and 54% at 250 and 700 W m(-2), respectively. However, the amount of quinclorac in paddy field water remained almost constant under the same irradiation conditions. QNC dissipated completely after 40 min of TiO(2) photocatalysis in ultrapure water, whereas 130 min were necessary to degrade 98% of the initial concentration in paddy field water. Possible QNC photolytic and photocatalytic degradation pathways are proposed after structure elucidation of the main transformation products, through liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry and exact mass measurements. Pyridine ring hydroxylation at C-9 followed by ring opening and/or oxidative dechlorination were the key steps of QNC degradation. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. Cometabolic degradation of chloramphenicol via a meta-cleavage pathway in a microbial fuel cell and its microbial community.

    Science.gov (United States)

    Zhang, Qinghua; Zhang, Yanyan; Li, Daping

    2017-04-01

    The performance of a microbial fuel cell (MFC) in terms of degradation of chloramphenicol (CAP) was investigated. Approximately 84% of 50mg/L CAP was degraded within 12h in the MFC. A significant interaction of pH, temperature, and initial CAP concentration was found on removal of CAP, and a maximum degradation rate of 96.53% could theoretically be achieved at 31.48°C, a pH of 7.12, and an initial CAP concentration of 106.37mg/L. Moreover, CAP was further degraded through a ring-cleavage pathway. The antibacterial activity of CAP towards Escherichia coli ATCC 25922 and Shewanella oneidensis MR-1 was largely eliminated by MFC treatment. High-throughput sequencing analysis indicated that Azonexus, Comamonas, Nitrososphaera, Chryseobacterium, Azoarcus, Rhodococcus, and Dysgonomonas were the predominant genera in the MFC anode biofilm. In conclusion, the MFC shows potential for the treatment of antibiotic residue-containing wastewater due to its high rates of CAP removal and energy recovery.

  9. Gamma radiolytic eradication of methoxychlor in aqueous media. The degradation pathways using HPLC and SPME-GC-MS

    Energy Technology Data Exchange (ETDEWEB)

    Butt, S.B.; Zafar, A. [PINSTECH, Nilore, Islamabad (Pakistan). Central Analytical Facility Div.; Riaz, M. [PINSTECH, Nilore, Islamabad (Pakistan). Chemistry Div.

    2013-08-01

    The gamma radiation-induced degradation of environmental pollutant methoxychlor in water was investigated. A {sup 60}Co gamma radiation source with a dose rate of 372 Gy h{sup -1} was used for gamma irradiation of 1 mg L{sup -1} and 10 mg L{sup -1} methoxychlor in water with a varied absorbed dose of 1-5 kGy. A single step clean up and pre-concentration procedure based on solid phase micro-extraction was optimized. The extent of radiolytic degradation was monitored by reversed phase HPLC-UV and GC-ECD. The trace and ultra trace level degradation products were identified using GC-MS-SPME by comparing their mass spectra with the NIST 98 m mass spectral library. Most of the generated products for 4 kGy dose are substituted chlorophenols. The reaction pathways of these substituted chlorophenols and benzophenone formation are also proposed. However, generated chlorophenols disappeared along with methoxychlor for an absorbed dose of 5 kGy. The attained degradation of methoxychlor is {proportional_to} 95% that reflects the potential use of ionization radiation for wastewater treatment. (orig.)

  10. Identification and characterization of stress degradants of lacosamide by LC-MS and ESI-Q-TOF-MS/MS: development and validation of a stability indicating RP-HPLC method.

    Science.gov (United States)

    Ramisetti, Nageswara Rao; Kuntamukkala, Ramakrishna; Lakshetti, Sridhar; Sripadi, Prabhakar

    2014-07-01

    The current study dealt with the degradation behavior of lacosamide (LAC) under ICH prescribed stress conditions. LAC was found to be labile under acid and base hydrolytic stress conditions, while it was stable to neutral hydrolytic, oxidative, photolytic and thermal stress. In total, seven degradation products (DPs) were formed, which were separated on a C18 column using a stability-indicating method. LC-MS analyses indicated that one of the DPs had the same molecular mass as that of the drug. Structural characterization of DPs was carried out using ESI-Q-TOF-MS/MS technique. The degradation pathways and mechanisms of degradation of the drug were delineated by carrying out the degradation in different co-solvents viz. methanol, deuterated methanol, ethanol, 1-propanol and acetonitrile. The developed LC method was validated for the determination of related substances and assay of LAC as per ICH guidelines. This study demonstrates a comprehensive approach of LAC degradation studies during its development phase.

  11. Sector-wise midpoint characterization factors for impact assessment of regional consumptive and degradative water use.

    Science.gov (United States)

    Lin, Chia-Chun; Lin, Jia-Yu; Lee, Mengshan; Chiueh, Pei-Te

    2017-12-31

    Water availability, resulting from either a lack of water or poor water quality is a key factor contributing to regional water stress. This study proposes a set of sector-wise characterization factors (CFs), namely consumptive and degradative water stresses, to assess the impact of water withdrawals with a life cycle assessment approach. These CFs consider water availability, water quality, and competition for water between domestic, agricultural and industrial sectors and ecosystem at the watershed level. CFs were applied to a case study of regional water management of industrial water withdrawals in Taiwan to show that both regional or seasonal decrease in water availability contributes to a high consumptive water stress, whereas water scarcity due to degraded water quality not meeting sector standards has little influence on increased degradative water stress. Degradative water stress was observed more in the agricultural sector than in the industrial sector, which implies that the agriculture sector may have water quality concerns. Reducing water intensity and alleviating regional scale water stresses of watersheds are suggested as approaches to decrease the impact of both consumptive and degradative water use. The results from this study may enable a more detailed sector-wise analysis of water stress and influence water resource management policies. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Isolation and characterization of novel atrazine-degrading microorganisms from an agricultural soil.

    Science.gov (United States)

    Vibber, Laurel L; Pressler, Michael J; Colores, Gregory M

    2007-06-01

    Six previously undescribed microorganisms capable of atrazine degradation were isolated from an agricultural soil that received repeated exposures of the commonly used herbicides atrazine and acetochlor. These isolates are all Gram-positive and group with microorganisms in the genera Nocardioides and Arthrobacter, both of which contain previously described atrazine degraders. All six isolates were capable of utilizing atrazine as a sole nitrogen source when provided with glucose as a separate carbon source. Under the culture conditions used, none of the isolates could utilize atrazine as the sole carbon and nitrogen source. We used several polymerase-chain-reaction-based assays to screen for the presence of a number of atrazine-degrading genes and verified their identity through sequencing. All six isolates contain trzN and atzC, two well-characterized genes involved in the conversion of atrazine to cyanuric acid. An additional atrazine-degrading gene, atzB, was detected in one of the isolates as well, yet none appeared to contain atzA, a commonly encountered gene in atrazine impacted soils and atrazine-degrading isolates. Interestingly, the deoxyribonucleic acid sequences of trzN and atzC were all identical, implying that their presence may be the result of horizontal gene transfer among these isolates.

  13. Identification and characterization of humic substances-degrading bacterial isolates from an estuarine environment.

    Science.gov (United States)

    Esham; Ye; Moran

    2000-12-01

    Bacterial isolates were obtained from enrichment cultures containing humic substances extracted from estuarine water using an XAD-8 resin. Eighteen isolates were chosen for phylogenetic and physiological characterization based on numerical importance in serial dilutions of the enrichment culture and unique colony morphology. Partial sequences of the 16S rRNA genes indicated that six of the isolates were associated with the alpha subclass of Proteobacteria, three with the gamma-Proteobacteria, and nine with the Gram-positive bacteria. Ten isolates degraded at least one (and up to six) selected aromatic single-ring compounds. Six isolates showed ability to degrade [(14)C]humic substances derived from the dominant salt marsh grass in the estuary from which they were isolated (Spartina alterniflora), mineralizing 0.4-1.1% of the humic substances over 4 weeks. A mixture of all 18 isolates did not degrade humic substances significantly faster than any of the individual strains, however, and no isolate degraded humic substances to the same extent as the natural marine bacterial community (3.0%). Similar studies with a radiolabeled synthetic lignin ([beta-(14)C]dehydropolymerisate) showed measurable levels of degradation by all 18 bacteria (3.0-8.8% in 4 weeks), but mineralization levels were again lower than that observed for the natural marine bacterial community (28.2%). Metabolic capabilities of the 18 isolates were highly variable and generally did not map to phylogenetic affiliation.

  14. Novel Chromatic Technique Based on Optical Absorbance in Characterizing Mineral Hydraulic Oil Degradation

    Directory of Open Access Journals (Sweden)

    C. V. Ossia

    2012-01-01

    Full Text Available A low cost, compact, real-time, and quick measurement optical device based on the absorbance of white light, which comprised of photodiodes in a 3-element color-sensor, feedback diodes, water and temperature sensing element, and so on, was developed and tested in low absorption mineral oil. The device, a deviation from conventional electrical, mechanical, and electrochemical techniques, uses color ratio (CR and total contamination index (TCI parameters based on transmitted light intensity in RGB wavelengths for oil condition monitoring. Test results showed that CR corroborated CIE chromaticity (- Coordinates and increased with oil degradation unlike Saturation and Hue . CR was found to be independent of the particulate contaminants of oil, but dependent on chemical degradation. TCI depended on both chemical degradation and particulate contaminants in oil, being most sensitive in the blue wavelength range and least in the green. Furthermore, results agreed with those of viscometry, total acid number (TAN, and UV-VIS photospectrometry. CR and TCI gave clearer indication of oil degradation than key monitoring parameters like TAN and were found to be effective criteria for characterizing the degradation of hydraulic mineral oils.

  15. Identification, Characterization, and Quantification of Impurities of Safinamide Mesilate: Process-Related Impurities and Degradation Products.

    Science.gov (United States)

    Zou, Liang; Sun, Lili; Zhang, Hui; Hui, Wenkai; Zou, Qiaogen; Zhu, Zheying

    2017-07-01

    The characterization of process-related impurities and degradation products of safinamide mesilate (SAFM) in bulk drug and a stability-indicating HPLC method for the separation and quantification of all the impurities were investigated. Four process-related impurities (Imp-B, Imp-C, Imp-D, and Imp-E) were found in the SAFM bulk drug. Five degradation products (Imp-A, Imp-C, Imp-D, Imp-E, and Imp-F) were observed in SAFM under oxidative conditions. Imp-C, Imp-D, and Imp-E were also degradation products and process-related impurities. Remarkably, one new compound, identified as (S)-2-[4-(3-fluoro-benzyloxy) benzamido] propanamide (i.e., Imp-D), is being reported here as an impurity for the first time. Furthermore, the structures of the aforementioned impurities were characterized and confirmed via IR, NMR, and MS techniques, and the most probable formation mechanisms of all impurities proposed according to the synthesis route. Optimum separation was achieved on an Inertsil ODS-3 column (250 × 4.6 mm, 5 μm), using 0.1% formic acid in water (pH adjusted to 5.0) and acetonitrile as the mobile phase in gradient mode. The proposed method was found to be stability-indicating, precise, linear, accurate, sensitive, and robust for the quantitation of SAFM and its process-related substances, including its degradation products.

  16. Characterization of Chlorinated Ethene Degradation in a Vertical Flow Constructed Wetland

    Science.gov (United States)

    2007-03-01

    pathway for chlorinated volatiles in phytoremediation applications. Although transpiration of chlorinated solvents has been confirmed in studies ... case study publications and conference presentations providing support for the use of constructed wetlands for the treatment of chlorinated solvent...groundwater. This study characterized and evaluated the concentration of chlorinated ethenes within a vertical flow constructed wetland, fed with PCE

  17. Selective separation, detection of zotepine and mass spectral characterization of degradants by LC-MS/MS/QTOF

    Institute of Scientific and Technical Information of China (English)

    M.V.N. Kumar Talluri; Naveen Reddy Kandimalla; Raju Bandu; Divya Chundi; Ramesh Marupaka; Ragampeta Srinivas

    2014-01-01

    A simple, precise, accurate stability-indicating gradient reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed for the quantitative determination of zotepine (ZTP) in bulk and pharmaceutical dosage forms in the presence of its degradation products (DPs). The method was developed using Phenomenex C18 column (250 mm ~ 4.6 mm i.d., 5 mm) with a mobile phase containing a gradient mixture of solvents, A (0.05%trifluoroacetic acid (TFA), pH ¼ 3.0) and B (acetonitrile). The eluted compounds were monitored at 254 nm;the run time was within 20.0 min, in which ZTP and its DPs were well separated, with a resolution of 41.5. The stress testing of ZTP was carried out under acidic, alkaline, neutral hydrolysis, oxidative, photolytic and thermal stress conditions. ZTP was found to degrade significantly in acidic, photolytic, thermal and oxidative stress conditions and remain stable in basic and neutral conditions. The developed method was validated with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness as per ICH guidelines. This method was also suitable for the assay determination of ZTP in pharmaceutical dosage forms. The DPs were characterized by LC-MS/MS and their fragmentation pathways were proposed.

  18. Selective separation, detection of zotepine and mass spectral characterization of degradants by LC–MS/MS/QTOF

    Directory of Open Access Journals (Sweden)

    M.V.N. Kumar Talluri

    2014-04-01

    Full Text Available A simple, precise, accurate stability-indicating gradient reversed-phase high-performance liquid chromatographic (RP–HPLC method was developed for the quantitative determination of zotepine (ZTP in bulk and pharmaceutical dosage forms in the presence of its degradation products (DPs. The method was developed using Phenomenex C18 column (250 mm×4.6 mm i.d., 5 µm with a mobile phase containing a gradient mixture of solvents, A (0.05% trifluoroacetic acid (TFA, pH=3.0 and B (acetonitrile. The eluted compounds were monitored at 254 nm; the run time was within 20.0 min, in which ZTP and its DPs were well separated, with a resolution of >1.5. The stress testing of ZTP was carried out under acidic, alkaline, neutral hydrolysis, oxidative, photolytic and thermal stress conditions. ZTP was found to degrade significantly in acidic, photolytic, thermal and oxidative stress conditions and remain stable in basic and neutral conditions. The developed method was validated with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness as per ICH guidelines. This method was also suitable for the assay determination of ZTP in pharmaceutical dosage forms. The DPs were characterized by LC–MS/MS and their fragmentation pathways were proposed.

  19. Cloning, characterization and analysis of cat and ben genes from the phenol degrading halophilic bacterium Halomonas organivorans.

    Directory of Open Access Journals (Sweden)

    Maria de Lourdes Moreno

    Full Text Available BACKGROUND: Extensive use of phenolic compounds in industry has resulted in the generation of saline wastewaters that produce significant environmental contamination; however, little information is available on the degradation of phenolic compounds in saline conditions. Halomonas organivorans G-16.1 (CECT 5995(T is a moderately halophilic bacterium that we isolated in a previous work from saline environments of South Spain by enrichment for growth in different pollutants, including phenolic compounds. PCR amplification with degenerate primers revealed the presence of genes encoding ring-cleaving enzymes of the β-ketoadipate pathway for aromatic catabolism in H. organivorans. FINDINGS: The gene cluster catRBCA, involved in catechol degradation, was isolated from H. organivorans. The genes catA, catB, catC and the divergently transcribed catR code for catechol 1,2-dioxygenase (1,2-CTD, cis,cis-muconate cycloisomerase, muconolactone delta-isomerase and a LysR-type transcriptional regulator, respectively. The benzoate catabolic genes (benA and benB are located flanking the cat genes. The expression of cat and ben genes by phenol and benzoic acid was shown by RT-PCR analysis. The induction of catA gene by phenol and benzoic acid was also probed by the measurement of 1,2-CTD activity in H. organivorans growth in presence of these inducers. 16S rRNA and catA gene-based phylogenies were established among different degrading bacteria showing no phylogenetic correlation between both genes. CONCLUSIONS/SIGNIFICANCE: In this work, we isolated and determined the sequence of a gene cluster from a moderately halophilic bacterium encoding ortho-pathway genes involved in the catabolic metabolism of phenol and analyzed the gene organization, constituting the first report characterizing catabolic genes involved in the degradation of phenol in moderate halophiles, providing an ideal model system to investigate the potential use of this group of extremophiles in

  20. Identification, characterization and in silico ADMET prediction of Roflumilast degradation products.

    Science.gov (United States)

    Pinheiro, Mariana S; Viana, Gil M; Vieira, Bárbara de A Abrahim; de Souza, Alessandra Mendonça Teles; Rodrigues, Carlos Rangel; Marins, Rita de Cássia E E; Cabral, Lúcio M; de Sousa, Valéria P

    2017-05-10

    The present study reports the degradation behavior of roflumilast (RFL), a new drug developed for the treatment of chronic obstructive pulmonary disease. The degradation of RFL was tested under various stress conditions as per the guidelines of the International Conference on Harmonization. The degradation products (DPs) of RFL were identified, characterized and in silico predictions were made of their pharmacokinetic properties, absorption, distribution, metabolism, excretion and toxicity (ADMET). RFL was subjected to various stress conditions including photodegradation, alkaline and acidic hydrolysis, oxidative and metallic degradation. After analysis by HPLC-DAD, the DPs were isolated by preparative TLC and characterized by high resolution mass spectrometry (HRMS), (1)H NMR, (13)C NMR and infrared (IR) spectroscopy. RFL tablets were prepared by the addition of solid stressing substances such as excipients and storage in an accelerated stability chamber (40°C; 75% r.h.) for sixteen months. Resulting DPs from the tablets were analyzed by UFLC-QTOF. The most drastic degradation conditions for RFL were 5M NaOH(aq), 6M HCl(aq), 7.5% v/v peracetic acid, which resulted in the isolation of four DPs. However, milder degradation conditions (1M NaOH(aq) and photolysis) generated six DPs (DP-1, 2, 3, 5, 7 and 8), and are more similar to the actual conditions the drug will be exposed. For tablets containing RFL exposed to an alkaline reagent, two DPs were formed: DP-1 and DP-11. Whereas RFL-containing tablets exposed to acid and oxidizing agents, formed one product DP-11. Forced degradation of RFL led to the formation of eleven DPs, seven of which have never been previously reported. RFL is stable under metallic stress and it is relatively stable during photodegradation testing. The UFLC-QTOF methodology detected a greater number of DPs that formed during the stress conditions tested when compared to the HPLC-DAD methodology. In silico prediction of the ADMET properties of

  1. UV photolysis of diclofenac in water; kinetics, degradation pathway and environmental aspects.

    Science.gov (United States)

    Kovacic, Marin; Juretic Perisic, Daria; Biosic, Martina; Kusic, Hrvoje; Babic, Sandra; Loncaric Bozic, Ana

    2016-08-01

    In this study, the photolysis behavior of commonly used anti-inflammatory drug diclofenac (DCF) was investigated using UV-C and UV-A irradiation. In that purpose, DCF conversion kinetics, mineralization of organic content, biodegradability, and toxicity were monitored and compared. The results showed different kinetics of DCF conversion regarding the type of UV source applied. However, in both cases, the mineralization extent reached upon complete DCF conversion is rather low (≤10 %), suggesting that the majority of DCF was transformed into by-products. Formation/degradation of main degradation by-products was monitored using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS), whereas different profiles were obtained by UV-C and UV-A photolysis. The results of bioassays revealed that biodegradability of DCF solutions remained low through the applied treatments. The toxicity of irradiated DCF solutions was evaluated using Vibrio fischeri. A significant reduction of toxicity, especially in the case of UV-A radiation, was observed upon complete degradation of DCF. In addition to toxicity reduction, calculated Log K OW values of DCF degradation by-products indicate their low potential for bioaccumulation (Log K OW ≤ 3) in comparison to the parent substance.

  2. Peroxisome degradation in Saccharomyces cerevisiae is dependent on machinery of macroautophagy and the Cvt pathway

    NARCIS (Netherlands)

    Hutchins, Maria U.; Veenhuis, Marten; Klionsky, Daniel J.

    1999-01-01

    Organelle biogenesis and turnover are necessary to maintain biochemical processes that are appropriate to the needs of the eukaryotic cell. Specific degradation of organelles in response to changing environmental cues is one aspect of achieving proper metabolic function. For example, the yeast Sacch

  3. Peroxisome degradation in Saccharomyces cerevisiae is dependent on machinery of macroautophagy and the Cvt pathway

    NARCIS (Netherlands)

    Hutchins, Maria U.; Veenhuis, Marten; Klionsky, Daniel J.

    1999-01-01

    Organelle biogenesis and turnover are necessary to maintain biochemical processes that are appropriate to the needs of the eukaryotic cell. Specific degradation of organelles in response to changing environmental cues is one aspect of achieving proper metabolic function. For example, the yeast Sacch

  4. Stability of 6:2 fluorotelomer sulfonate in advanced oxidation processes: degradation kinetics and pathway.

    Science.gov (United States)

    Yang, Xiaoling; Huang, Jun; Zhang, Kunlun; Yu, Gang; Deng, Shubo; Wang, Bin

    2014-03-01

    Perfluorooctane sulfonate (PFOS), a widely used mist suppressant in hard chrome electroplating industry, has been listed in the Stockholm Convention for global ban. 6:2 Fluorotelomer sulfonate (6:2 FTS) acid and salts have been adopted as alternative products in the market, but no data about their abiotic degradation has been reported. In the present study, the degradability of 6:2 FTS potassium salt (6:2 FTS-K) was evaluated under various advanced oxidation processes, including ultraviolet (UV) irradiation, UV with hydrogen peroxide (H2O2), alkaline ozonation (O3, pH = 11), peroxone (O3/H2O2), and Fenton reagent oxidation (Fe(2+)/H2O2). UV/H2O2 was found to be the most effective approach, where the degradation of 6:2 FTS-K followed the pseudo-first-order kinetics. The intermediates were mainly shorter chain perfluoroalkyl carboxylic acid (C7 to C2), while sulfate (SO4 (2-)) and fluoride (F(-)) were found to be the final products. The high yields of SO4 (2-) and F(-) indicate that 6:2 FTS-K can be nearly completely desulfonated and defluorinated under UV/H2O2 condition. The degradation should firstly begin with the substitution of hydrogen atom by hydroxyl radicals, followed by desulfonation, carboxylation, and sequential "flake off" of CF2 unit. Compared with PFOS which is inert in most advanced oxidation processes, 6:2 FTS-K is more degradable as the alternative.

  5. Characterization of Biosurfactant Produced during Degradation of Hydrocarbons Using Crude Oil As Sole Source of Carbon

    Science.gov (United States)

    Patowary, Kaustuvmani; Patowary, Rupshikha; Kalita, Mohan C.; Deka, Suresh

    2017-01-01

    Production and spillage of petroleum hydrocarbons which is the most versatile energy resource causes disastrous environmental pollution. Elevated oil degrading performance from microorganisms is demanded for successful microbial remediation of those toxic pollutants. The employment of biosurfactant-producing and hydrocarbon-utilizing microbes enhances the effectiveness of bioremediation as biosurfactant plays a key role by making hydrocarbons bio-available for degradation. The present study aimed the isolation of a potent biosurfactant producing indigenous bacteria which can be employed for crude oil remediation, along with the characterization of the biosurfactant produced during crude oil biodegradation. A potent bacterial strain Pseudomonas aeruginosa PG1 (identified by 16s rDNA sequencing) was isolated from hydrocarbon contaminated soil that could efficiently produce biosurfactant by utilizing crude oil components as the carbon source, thereby leading to the enhanced degradation of the petroleum hydrocarbons. Strain PG1 could degrade 81.8% of total petroleum hydrocarbons (TPH) after 5 weeks of culture when grown in mineral salt media (MSM) supplemented with 2% (v/v) crude oil as the sole carbon source. GCMS analysis of the treated crude oil samples revealed that P. aeruginosa PG1 could potentially degrade various hydrocarbon contents including various PAHs present in the crude oil. Biosurfactant produced by strain PG1 in the course of crude oil degradation, promotes the reduction of surface tension (ST) of the culture medium from 51.8 to 29.6 mN m−1, with the critical micelle concentration (CMC) of 56 mg L−1. FTIR, LC-MS, and SEM-EDS studies revealed that the biosurfactant is a rhamnolipid comprising of both mono and di rhamnolipid congeners. The biosurfactant did not exhibit any cytotoxic effect to mouse L292 fibroblastic cell line, however, strong antibiotic activity against some pathogenic bacteria and fungus was observed. PMID:28275373

  6. 13C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H

    Directory of Open Access Journals (Sweden)

    Steffen Ostermann

    2015-09-01

    Full Text Available Gluconobacter oxydans 621H is used as an industrial production organism due to its exceptional ability to incompletely oxidize a great variety of carbohydrates in the periplasm. With glucose as the carbon source, up to 90% of the initial concentration is oxidized periplasmatically to gluconate and ketogluconates. Growth on glucose is biphasic and intracellular sugar catabolism proceeds via the Entner–Doudoroff pathway (EDP and the pentose phosphate pathway (PPP. Here we studied the in vivo contributions of the two pathways to glucose catabolism on a microtiter scale. In our approach we applied specifically 13C labeled glucose, whereby a labeling pattern in alanine was generated intracellularly. This method revealed a dynamic growth phase-dependent pathway activity with increased activity of EDP in the first and PPP in the second growth phase, respectively. Evidence for a growth phase-independent decarboxylation-carboxylation cycle around the pyruvate node was obtained from 13C fragmentation patterns of alanine. For the first time, down-scaled microtiter plate cultivation together with 13C-labeled substrate was applied for G. oxydans to elucidate pathway operation, exhibiting reasonable labeling costs and allowing for sufficient replicate experiments.

  7. Characterization of Degradation Progressive in Composite Laminates Subjected to Thermal Fatigue and Moisture Diffusion by Lamb Waves

    Directory of Open Access Journals (Sweden)

    Weibin Li

    2016-02-01

    Full Text Available Laminate composites which are widely used in the aeronautical industry, are usually subjected to frequency variation of environmental temperature and excessive humidity in the in-service environment. The thermal fatigue and moisture absorption in composites may induce material degradation. There is a demand to investigate the coupling damages mechanism and characterize the degradation evolution of composite laminates for the particular application. In this paper, the degradation evolution in unidirectional carbon/epoxy composite laminates subjected to thermal fatigue and moisture absorption is characterized by Lamb waves. The decrease rate of Lamb wave velocity is used to track the degradation evolution in the specimens. The results show that there are two stages for the progressive degradation of composites under the coupling effect of thermal cyclic loading and moisture diffusion. The present work provides an alternative to monitoring the degradation evolution of in-service aircraft composite Laminates.

  8. Characterization of Degradation Progressive in Composite Laminates Subjected to Thermal Fatigue and Moisture Diffusion by Lamb Waves.

    Science.gov (United States)

    Li, Weibin; Xu, Chunguang; Cho, Younho

    2016-01-01

    Laminate composites which are widely used in the aeronautical industry, are usually subjected to frequency variation of environmental temperature and excessive humidity in the in-service environment. The thermal fatigue and moisture absorption in composites may induce material degradation. There is a demand to investigate the coupling damages mechanism and characterize the degradation evolution of composite laminates for the particular application. In this paper, the degradation evolution in unidirectional carbon/epoxy composite laminates subjected to thermal fatigue and moisture absorption is characterized by Lamb waves. The decrease rate of Lamb wave velocity is used to track the degradation evolution in the specimens. The results show that there are two stages for the progressive degradation of composites under the coupling effect of thermal cyclic loading and moisture diffusion. The present work provides an alternative to monitoring the degradation evolution of in-service aircraft composite Laminates.

  9. Pathways of nitrobenzene degradation in horizontal subsurface flow constructed wetlands: Effect of intermittent aeration and glucose addition.

    Science.gov (United States)

    Kirui, Wesley K; Wu, Shubiao; Kizito, Simon; Carvalho, Pedro N; Dong, Renjie

    2016-01-15

    Intermittent aeration and addition of glucose were applied to horizontal subsurface flow constructed wetlands in order to investigate the effect on pathways of nitrobenzene (NB) degradation and interactions with microbial nitrogen and sulphur transformations. The experiment was carried out in three phases A, B and C consisting of different NB loading and glucose dosing. For each phase, the effect of aeration was assessed by intermittently aerating one wetland and leaving one unaerated. Regardless of whether or not the wetland was aerated, at an influent NB concentration of 140 mg/L, both wetlands significantly reduced NB to less than 2 mg/L, a reduction efficiency of 98%. However, once the influent NB concentration was increased to 280 mg/L, the aerated wetland had a higher removal performance 82% compared to that of the unaerated wetland 71%. Addition of glucose further intensified the NB removal to 95% in the aerated wetlands and 92% in the unaerated. Aeration of wetlands enhanced NB degradation, but also resulted in higher NB volatilization of 6 mg m(-2) d(-1). The detected high concentration of sulphide 20-60 mg/L in the unaerated wetland gave a strong indication that NB may act as an electron donor to sulphate-reducing bacteria, but this should be further investigated. Aeration positively improved NB removal in constructed wetlands, but resulted in higher NB volatilization. Glucose addition induced co-metabolism to enhance NB degradation.

  10. Electrochemical degradation of sulfonamides at BDD electrode: Kinetics, reaction pathway and eco-toxicity evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Fabiańska, Aleksandra; Białk-Bielińska, Anna; Stepnowski, Piotr [Faculty of Chemistry, University of Gdansk, ul. Wita Stwosza 63, 80-952 Gdansk (Poland); Stolte, Stefan [Faculty of Chemistry, University of Gdansk, ul. Wita Stwosza 63, 80-952 Gdansk (Poland); UFT-Centre of Environmental Research and Sustainable Technology, University of Bremen, Leobener Straße UFT, D-28359 Bremen (Germany); Siedlecka, Ewa Maria, E-mail: ewa.siedlecka@ug.edu.pl [Faculty of Chemistry, University of Gdansk, ul. Wita Stwosza 63, 80-952 Gdansk (Poland)

    2014-09-15

    Highlights: • SNs were electrochemically oxidized at BDD in one compartment reactor. • The efficiency of SN degradation was the highest in effluents from municipal WWTP. • The electro-degradation SNs based on oxidation but reduction was also possible. • Electrochemical oxidation of SNs led in some cases to mixtures toxic to L. minor. - Abstract: The investigation dealt with electrochemical oxidation of five sulfonamides (SNs): sulfadiazine (SDZ), sulfathiazole (STZ), sulfamerazine (SMR), sulfamethazine (SMN) and sulfadimethoxine (SDM) in aqueous solution at boron-doped diamond (BDD) electrode. All studied sulfonamides were degraded according to a pseudo first order kinetics. The structure of SNs had no significant effect on the values of pseudo first order rate constants. Increased degradation efficiency was observed in higher temperature and in acidic pH. Due to the presence of chlorine and nitrate SNs were more effectively oxidized from municipal wastewater treatment plant (WWTP) effluents than from pure supporting electrolyte Na{sub 2}SO{sub 4}. The intermediates identified by LC–MS and GC–MS analysis suggested that the hydroxyl radicals attack mainly the S-N bond, but also the aromatic ring systems (aniline, pyrimidine or triazole) of SNs. Finally, the toxicity of the SNs solutions and effluents after electrochemical treatment was assessed through the measurement of growth inhibition of green algae (Scenedesmus vacualatus) and duckweed (Lemna minor). Toxicity of SMR, STZ, SMN solutions before and after electrochemical oxidation and SDM solution after the process in L. minor test was observed. No significant toxicity of studied SNs was observed in algae test.

  11. Coupled Metabolic and Photolytic Pathway for Degradation of Pyridinedicarboxylic Acids, Especially Dipicolinic Acid

    OpenAIRE

    Amador, José A.; Taylor, Barrie F.

    1990-01-01

    Three isomers of pyridinedicarboxylic acid (PDCA) (2,3-, 2,5-, and 2,6-PDCA) were partially oxidized by marine bacteria when grown aerobically on the corresponding phthalate analogs. The metabolites, unlike the parent PDCAs, absorbed light in the solar actinic range (wavelengths greater than 300 nm) and were readily degraded in sunlight. The principal product from 2,6-PDCA (dipicolinic acid) metabolism was extracted from a culture fluid, purified by column chromatography, and analyzed by UV-v...

  12. Synthesis, Characterization, and Use of Novel Bimetal Oxide Catalyst for Photoassisted Degradation of Malachite Green Dye

    Directory of Open Access Journals (Sweden)

    K. L. Ameta

    2014-01-01

    Full Text Available This work reports a simple, novel, and cost effective synthesis of nanobimetal oxide catalyst using cerium and cadmium nitrates as metal precursors. The cerium-cadmium oxide nanophotocatalyst was synthesized by coprecipitation method and characterized by X-ray powder diffraction method to analyze the particle size. XRD study reveals a high degree of crystallinity and 28.43 nm particle size. The photocatalytic efficiency of the synthesized nanobimetal catalyst was examined by using it for the photocatalytic degradation of malachite green dye. Experiments were conducted to study the effect of various parameters, such as the pH of the dye solution, concentration of dye, amount of catalyst, and light intensity on the rate of dye degradation. The progress of the dye degradation was monitored spectrophotometrically by taking the optical density of the dye solution at regular intervals. Experimental results indicate that the dye degrades best at pH 8.0 with light intensity 600 Wm−2 and catalyst loading 0.03 g/50 mL of dye solution. The rate constant for the reaction was 7.67 × 10−4 s−1.

  13. Isolation, characterization and phylogenetic analysis of a bacterial strain capable of degrading acetamiprid

    Institute of Scientific and Technical Information of China (English)

    YAO Xiao-hua; MIN Hang

    2006-01-01

    An aerobic bacterium, capable of degrading the new chloronicotine pesticide acetamiprid, was isolated from the sludge of pesticide factory after successive enrichment cultures and named strain FH2 which is a Gram-negative, rod-shaped, obligate aerobic organism with ((0.5-0.7) ×(1.5-3.0))μm of cell size and with monotrichous flagellum. It was identified as a member of Pseudomonas sp. based on morphology, physio-biochemical properties, Biolog GN2, 16S rDNA sequence and phylogenetic characteristic analysis. The isolate could grow optimally at pH 7.0 and 30℃ in acetamiprid-mineral medium with 800 mg/L concentration. About 53.3% acetamiprid was degraded by strain FH2 after incubation for 14 d in acetamiprid-mineral medium and nearly 96.7% degraded when incubated in acetamiprid-yeast mineral medium at 30℃ for 14 d. This paper describes phylogenetic and degradation characterization of a pure bacterium being able to mineralize acetamiprid for the first time.

  14. Characterizing pesticide sorption and degradation in microscale biopurification systems using column displacement experiments

    Energy Technology Data Exchange (ETDEWEB)

    Wilde, Tineke de [Laboratory of Crop Protection Chemistry, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium)], E-mail: tineke.dewilde@UGent.be; Mertens, Jan [Division Soil and Water Management, Faculty of Bioscience Engineering, K.U. Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee (Belgium); Simunek, Jirka [Department of Environmental Sciences, University of California, Riverside, CA (United States); Sniegowksi, Kristel; Ryckeboer, Jaak [Division Soil and Water Management, Faculty of Bioscience Engineering, K.U. Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee (Belgium); Jaeken, Peter [PCF-Royal Research Station of Gorsem, De Brede Akker 13, 3800 Sint-Truiden (Belgium); Springael, Dirk [Division Soil and Water Management, Faculty of Bioscience Engineering, K.U. Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee (Belgium); Spanoghe, Pieter [Laboratory of Crop Protection Chemistry, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium)

    2009-02-15

    Biopurification systems treating pesticide contaminated water are very efficient, however they operate as a black box. Processes inside the system are not yet characterized. To optimize the performance, knowledge of degradation and retention processes needs to be generated. Therefore, displacement experiments were carried out for four pesticides (isoproturon, bentazone, metalaxyl, linuron) in columns containing different organic mixtures. Bromide, isoproturon and bentazone breakthrough curves (BTCs) were well described using the convection-dispersion equation (CDE) and a first-order degradation kinetic approach. Metalaxyl and linuron BTCs were well described using the CDE model expanded with Monod-type kinetics. Freundlich sorption, first-order degradation and Monod kinetics coefficients were fitted to the BTCs. Fitted values of the distribution coefficient K{sub f,column} were much lower than those determined from batch experiments. Based on mobility, pesticides were ranked as: bentazone > metalaxyl - isoproturon > linuron. Based on degradability, pesticides were ranked as: linuron > metalaxyl - isoproturon > bentazone. - Transport of pesticides in column experiments containing organic substrates.

  15. Isolation, Characterization, and Degradation Performance of the 17β-Estradiol-Degrading Bacterium Novosphingobium sp. E2S

    Directory of Open Access Journals (Sweden)

    Shunyao Li

    2017-01-01

    Full Text Available A 17β-estradiol (E2-degrading bacterium E2S was isolated from the activated sludge in a sewage treatment plant (STP. The morphology, biological characteristics, and 16S ribosomal RNA (rRNA gene sequence of strain E2S indicated that it belonged to the genus Novosphingobium. The optimal degrading conditions were 30 °C and pH 7.0. The ideal inoculum volume was 5% (v/v, and a 20-mL degradation system was sufficient to support the removal ability of strain E2S. The addition of extra NaCl to the system did not benefit the E2 degradation in batch culture by this strain. Strain E2S exhibited high degradation efficiency with initial substrate concentrations of 10–50 mg·L−1. For example, in mineral salt medium containing 50 mg·L−1 of E2, the degradation efficiency was 63.29% after seven days. In cow manure samples supplemented with 50 mg·L−1 of E2, strain E2S exhibited 66.40% degradation efficiency after seven days. The finding of the E2-degrading strain E2S provided a promising method for removing E2 from livestock manure in order to reduce the potential environmental risks of E2.

  16. Enhanced catalytic degradation of ciprofloxacin with FeS2/SiO2 microspheres as heterogeneous Fenton catalyst: Kinetics, reaction pathways and mechanism.

    Science.gov (United States)

    Diao, Zeng-Hui; Xu, Xiang-Rong; Jiang, Dan; Li, Gang; Liu, Jin-Jun; Kong, Ling-Jun; Zuo, Lin-Zi

    2017-04-05

    In this study, the application of FeS2/SiO2 microspheres as a catalyst to activate H2O2 for the degradation of ciprofloxacin (CIP) was systematically investigated. Results demonstrated that the presence of SiO2 microspheres on the surface of FeS2 could effectively make the reaction of aqueous Fe(2+) and H2O2 smoothly continuous by controlling the release of aqueous Fe(2+) from FeS2. Nearly 100% of CIP was degraded after 60min under the optimum conditions. A superior performance on the CIP degradation and high reusability of the catalyst was obtained in FeS2/SiO2 microspheres activated H2O2 system. A low concentration of ethylene diamine tetraacetie acid (EDTA) did positively affect the degradation rate of CIP. A synergetic effect between adsorption and oxidation processes contributed to the significant enhancement of CIP degradation. Seven oxidation intermediates were identified during the CIP degradation process, and the direct HO oxidation proved to be a main CIP degradation pathway. For degradation pathway of CIP, oxidation of piperazine ring would be its first step, followed by cleavage of the heterocyclic ring. Subsequently, the substitution, hydroxylation and decarboxylation processes occurred. This is the first report on the feasibility of FeS2/SiO2 microspheres activated H2O2 system for the enhanced degradation of CIP. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Molecular characterization of Lactobacillus plantarum DMDL 9010, a strain with efficient nitrite degradation capacity.

    Science.gov (United States)

    Fei, Yong-tao; Liu, Dong-mei; Luo, Tong-hui; Chen, Gu; Wu, Hui; Li, Li; Yu, Yi-gang

    2014-01-01

    Nitrites commonly found in food, especially in fermented vegetables, are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. A Lactobacillus strain (Lactobacillus sp. DMDL 9010) was previously isolated from fermented vegetables by our group, and is not yet fully characterized. A number of phenotypical and genotypical approaches were employed to characterize Lactobacillus sp. DMDL 9010. Its nitrite degradation capacity was compared with four other Lactobacillus strains, including Lactobacillus casei subsp. rhamnosus 719, Lactobacillus delbrueckii subsp. bulgaricu 1.83, Streptococcus thermophilus 1.204, and lactobacillus plantarum 8140, on MRS medium. Compared to these four Lactobacillus strains, Lactobacillus sp. DMDL 9010 had a significantly higher nitrite degradation capacity (PLactobacillus sp. DMDL 9010 was identified as either Lactobacillus plantarum or Lactobacillus pentosus. To further identify this strain, the flanking regions (922 bp and 806 bp upstream and downstream, respectively) of the L-lactate dehydrogenase 1 (L-ldh1) gene were amplified and sequenced. Lactobacillus sp. DMDL 9010 had 98.92 and 76.98% sequence identity in the upstream region with L. plantarum WCFS1 and L. pentosus IG1, respectively, suggesting that Lactobacillu sp. DMDL 9010 is an L. plantarum strain. It was therefore named L. plantarum DMDL 9010. Our study provides a platform for genetic engineering of L. plantarum DMDL 9010, in order to further improve its nitrite degradation capacity.

  18. Decomposition of acetaminophen in water by a gas phase dielectric barrier discharge plasma combined with TiO2-rGO nanocomposite: Mechanism and degradation pathway

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Guyu; Sun, Yabing, E-mail: sybnju@163.com; Zhang, Chunxiao; Yu, Zhongqing

    2017-02-05

    Highlights: • Graphene Oxide-based catalyst was first applied with dielectric barrier discharge plasma. • The TiO{sub 2}-rGO showed efficient synergistic effect with gas phase dielectric barrier discharge plasma. • The property changes of TiO{sub 2}-rGO nanocomposite after plasma treatment were characterized. • The mechanism and possible pathways of APAP degradation in plasma/TiO{sub 2}-rGO system were proposed. - Abstract: Acetaminophen (APAP) served as the model pollutant to evaluate the feasibility of pollutant removal by gas phase dielectric barrier discharge plasma combined with the titanium dioxide-reduced Graphene Oxide (TiO{sub 2}-rGO) nanocomposite. TiO{sub 2}-rGO nanocomposite was prepared using the modified hydrothermal method and characterized by TEM and XPS before and after plasma process. The results indicated that the APAP degradation efficiency was significantly improved to 92% after 18 min of discharge plasma treatment coupling 0.25 g L{sup −1} TiO{sub 2}-rGO 5% wt at 18 kV, compared with the plasma alone and plasma combined with P25 TiO{sub 2}. The degradation mechanism for APAP in this system was studied by investigating the effects of the operational variables (e.g. discharge voltage and pH value) and the amount of the generated active species; and the results showed that O{sub 3} and H{sub 2}O{sub 2} yields were influenced notably by adding TiO{sub 2}-rGO. Also, it was observed that, compared with unused TiO{sub 2}-rGO, the photocatalytic performance of used TiO{sub 2}-rGO declined after several recirculation times due to the further reduction of Graphene Oxide in plasma system. Finally, intermediate products were analyzed by UV–vis spectrometry and HPLC/MS, and possible transformation pathways were identified with the support of theoretically calculating the frontier electron density of APAP.

  19. Degradation and drug release in calcium polyphosphate bioceramics: an MRI-based characterization.

    Science.gov (United States)

    Bray, J M; Filiaggi, M J; Bowen, C V; Beyea, S D

    2012-10-01

    Degradable, bioceramic bone implants made of calcium polyphosphate (CPP) hold potential for controlled release of therapeutic agents in the treatment of localized bone disease. Magnetic resonance imaging techniques for non-invasively mapping fluid distribution, T(1) and T(2) relaxation times and the apparent diffusion coefficient were performed in conjunction with a drug elution protocol to resolve free and bound water components within the material microstructure in two CPP formulations (G1 and G2). The T(2) maps provided the most accurate estimates of free and bound water, and showed that G1 disks contained a detectable free water component at all times, with drug release dominated by a Fickian diffusion mechanism. Drug release from G2 disks was characterized by a combined diffusional/structural relaxation mechanism, which may be related to the gradual infiltration of a free water component associated with swelling and/or chemical degradation.

  20. Kinetic study of olive oil degradation monitored by fourier transform infrared spectrometry. Application to oil characterization.

    Science.gov (United States)

    Román Falcó, Iván P; Grané Teruel, Nuria; Prats Moya, Soledad; Martín Carratalá, M Luisa

    2012-11-28

    A new approach for the determination of kinetic parameters of the cis/trans isomerization during the oxidation process of 24 virgin olive oils belonging to 8 different varieties is presented. The accelerated process of degradation at 100 °C was monitored by recording the Fourier transform infrared spectra. The parameters obtained confirm pseudo-first-order kinetics for the degradation of cis and the appearance of trans double bonds. The kinetic approach affords the induction time and the rate coefficient; these parameters are related to the fatty acid profile of the fresh olive oils. The data obtained were used to compare the oil stability of the samples with the help of multivariate statistical techniques. Fatty acid allowed a classification of the samples in five groups, one of them constituted by the cultivars with higher stability. Meanwhile, the kinetic parameters showed greater ability for the characterization of olive oils, allowing the classification in seven groups.

  1. Characterization of Rhodococcus wratislaviensis strain J3 that degrades 4-nitrocatechol and other nitroaromatic compounds.

    Science.gov (United States)

    Navrátilová, Jitka; Tvrzová, Ludmila; Durnová, Eva; Spröer, Cathrin; Sedlácek, Ivo; Neca, Jirí; Nemec, Miroslav

    2005-02-01

    The bacterial strain J3 was isolated from soil by selective enrichment on mineral medium containing 4-nitrocatechol as the sole carbon and energy source. This strain was identified as Rhodococcus wratislaviensis on the basis of morphology, biochemical, physiological and chemotaxonomic characterization and complete sequencing of the 16S rDNA gene. The isolated bacterium could utilize 4-nitrocatechol, 3-nitrophenol and 5-nitroguaiacol as sole carbon and energy sources. Stoichiometric release of nitrites was measured during degradation of 4-nitrocatechol both in growing cultures and for stationary phase cells. The J3 strain was unable to degrade 4-nitroguaiacol, 2-nitrophenol, 4-nitrophenol, 2,4-dinitrobenzoic acid, 4,5-dimethoxy-2-nitrobenzoic acid and 2,3-difluoro-6-nitrophenol. The J3 strain is deposited in the Czech Collection of Microorganisms as CCM 4930.

  2. Direct Synthesis, Characterization and Catalytic Performance of Iron-Containing SBA-15 for Phenol Degradation

    Institute of Scientific and Technical Information of China (English)

    XIE Huan-ling; XU Wen-guo

    2008-01-01

    An iron-containing SBA-15(Fe-SBA-15) has been synthesized via one-pot hydrothermal method under weak acidic conditions. A series of characterizations show nanocomposite materials of iron particles supported over mesostructured materials. The catalytic activity of these iron-containing SBA-15 materials has been tested for the heterogeneous Fenton degradation of phenolic aqueous solutions. The catalytic performance has been monitored in terms of phenol conversion, whereas the catalytic stability was evaluated by catalyst recycle. The influence of concentration of hydrogen peroxide, catalyst loading, catalyst prepared with different Fe/Si molar ratios in the gel and pH values of the solution on phenol conversion has been studied. Achieving a good catalytic performance accompanied with a noteworthy stability, Fe-SBA-15 materials prepared by this method are shown as the successful catalyst for degradation of phenolic aqueous solutions by Fenton process.

  3. Population sinks resulting from degraded habitats of an obligate life-history pathway.

    Science.gov (United States)

    Hickford, Michael J H; Schiel, David R

    2011-05-01

    Many species traverse multiple habitats across ecosystems to complete their life histories. Degradation of critical, life stage-specific habitats can therefore lead to population bottlenecks and demographic deficits in sub-populations. The riparian zone of waterways is one of the most impacted areas of the coastal zone because of urbanisation, deforestation, farming and livestock grazing. We hypothesised that sink populations can result from alterations of habitats critical to the early life stages of diadromous fish that use this zone, and tested this with field-based sampling and experiments. We found that for Galaxias maculatus, one of the most widely distributed fishes of the southern hemisphere, obligate riparian spawning habitat was very limited and highly vulnerable to disturbance across 14 rivers in New Zealand. Eggs were laid only during spring tides, in the highest tidally influenced vegetation of waterways. Egg survival increased to >90% when laid in three riparian plant species and where stem densities were great enough to prevent desiccation, compared to no survival where vegetation was comprised of other species or was less dense. Experimental exclusion of livestock, one of the major sources of riparian degradation in rural waterways, resulted in quick regeneration, a tenfold increase in egg laying by fish and a threefold increase in survival, compared to adjacent controls. Overall, there was an inverse relationship between river size and egg production. Some of the largest rivers had little or no spawning habitat and very little egg production, effectively becoming sink populations despite supporting large adult populations, whereas some of the smallest pristine streams produced millions of eggs. We demonstrate that even a wide-ranging species with many robust adult populations can be compromised if a stage-specific habitat required to complete a life history is degraded by localised or more diffuse impacts.

  4. Pathways and substrate-specific regulation of amino acid degradation in Phaeobacter inhibens DSM 17395 (archetype of the marine Roseobacter clade).

    Science.gov (United States)

    Drüppel, Katharina; Hensler, Michael; Trautwein, Kathleen; Koßmehl, Sebastian; Wöhlbrand, Lars; Schmidt-Hohagen, Kerstin; Ulbrich, Marcus; Bergen, Nils; Meier-Kolthoff, Jan P; Göker, Markus; Klenk, Hans-Peter; Schomburg, Dietmar; Rabus, Ralf

    2014-01-01

    Combining omics and enzymatic approaches, catabolic routes of nine selected amino acids (tryptophan, phenylalanine, methionine, leucine, isoleucine, valine, histidine, lysine and threonine) were elucidated in substrate-adapted cells of Phaeobacter inhibens DSM 17395 (displaying conspicuous morphotypes). The catabolic network [excluding tricarboxylic acid (TCA) cycle] was reconstructed from 71 genes (scattered across the chromosome; one-third newly assigned), with 69 encoded proteins and 20 specific metabolites identified, and activities of 10 different enzymes determined. For example, Ph. inhibens DSM 17395 does not degrade lysine via the widespread saccharopine pathway but might rather employ two parallel pathways via 5-aminopentanoate or 2-aminoadipate. Tryptophan degradation proceeds via kynurenine and 2-aminobenzoate; the latter is metabolized as known from Azoarcus evansii. Histidine degradation is analogous to the Pseudomonas-type Hut pathway via N-formyl-l-glutamate. For threonine, only one of the three genome-predicted degradation pathways (employing threonine 3-dehydrogenase) is used. Proteins of the individual peripheral degradation sequences in Ph. inhibens DSM 17395 were apparently substrate-specifically formed contrasting the non-modulated TCA cycle enzymes. Comparison of genes for the reconstructed amino acid degradation network in Ph. inhibens DSM 17395 across 27 other complete genomes of Roseobacter clade members revealed most of them to be widespread among roseobacters.

  5. Insulin-degrading enzyme secretion from astrocytes is mediated by an autophagy-based unconventional secretory pathway in Alzheimer disease.

    Science.gov (United States)

    Son, Sung Min; Cha, Moon-Yong; Choi, Heesun; Kang, Seokjo; Choi, Hyunjung; Lee, Myung-Shik; Park, Sun Ah; Mook-Jung, Inhee

    2016-05-01

    The secretion of proteins that lack a signal sequence to the extracellular milieu is regulated by their transition through the unconventional secretory pathway. IDE (insulin-degrading enzyme) is one of the major proteases of amyloid beta peptide (Aβ), a presumed causative molecule in Alzheimer disease (AD) pathogenesis. IDE acts in the extracellular space despite having no signal sequence, but the underlying mechanism of IDE secretion extracellularly is still unknown. In this study, we found that IDE levels were reduced in the cerebrospinal fluid (CSF) of patients with AD and in pathology-bearing AD-model mice. Since astrocytes are the main cell types for IDE secretion, astrocytes were treated with Aβ. Aβ increased the IDE levels in a time- and concentration-dependent manner. Moreover, IDE secretion was associated with an autophagy-based unconventional secretory pathway, and depended on the activity of RAB8A and GORASP (Golgi reassembly stacking protein). Finally, mice with global haploinsufficiency of an essential autophagy gene, showed decreased IDE levels in the CSF in response to an intracerebroventricular (i.c.v.) injection of Aβ. These results indicate that IDE is secreted from astrocytes through an autophagy-based unconventional secretory pathway in AD conditions, and that the regulation of autophagy is a potential therapeutic target in addressing Aβ pathology.

  6. In-Situ Characterization of Potential-Induced Degradation in Crystalline Silicon Photovoltaic Modules Through Dark I–V Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Luo, Wei; Hacke, Peter; Singh, Jai Prakash; Chai, Jing; Wang, Yan; Ramakrishna, Seeram; Aberle, Armin G.; Khoo, Yong Sheng

    2017-01-01

    A temperature correction methodology for in-situ dark I-V(DIV) characterization of conventional p-type crystalline silicon photovoltaic (PV) modules undergoing potential-induced degradation (PID) is proposed.

  7. Regulation of protein degradation pathways by amino acids and insulin in skeletal muscle of neonatal pigs

    Institute of Scientific and Technical Information of China (English)

    Agus Suryawan; Teresa ADavis

    2014-01-01

    Background:The rapid gain in lean mass in neonates requires greater rates of protein synthesis than degradation. We previously delineated the molecular mechanisms by which insulin and amino acids, especially leucine, modulate skeletal muscle protein synthesis and how this changes with development. In the current study, we identified mechanisms involved in protein degradation regulation. In experiment 1, 6-and 26-d-old pigs were studied during 1) euinsulinemic-euglycemic-euaminoacidemic, 2) euinsulinemic-euglycemic-hyperaminoacidemic, and 3) hyperinsulinemic-euglycemic-euaminoacidemic clamps for 2 h. In experiment 2, 5-d-old pigs were studied during 1) euinsulinemic-euglycemic-euaminoacidemic-euleucinemic, 2) euinsulinemic-euglycemic-hypoaminoacidemic-hyperleucinemic, and 3) euinsulinemic-euglycemic-euaminoacidemic-hyperleucinemic clamps for 24 h. We determined in muscle indices of ubiquitin-proteasome, i.e., atrogin-1 (MAFbx) and muscle RING-finger protein-1 (MuRF1) and autophagy-lysosome systems, i.e., unc51-like kinase 1 (UKL1), microtubule-associated protein light chain 3 (LC3), and lysosomal-associated membrane protein 2 (Lamp-2). For comparison, we measured ribosomal protein S6 (rpS6) and eukaryotic initiation factor 4E (eIF4E) activation, components of translation initiation. Results:Abundance of atrogin-1, but not MuRF1, was greater in 26-than 6-d-old pigs and was not affected by insulin, amino acids, or leucine. Abundance of ULK1 and LC3 was higher in younger pigs and not affected by treatment. The LC3-II/LC3-I ratio was reduced and ULK1 phosphorylation increased by insulin, amino acids, and leucine. These responses were more profound in younger pigs. Abundance of Lamp-2 was not affected by treatment or development. Abundance of eIF4E, but not rpS6, was higher in 6-than 26-d-old-pigs but unaffected by treatment. Phosphorylation of eIF4E was not affected by treatment, however, insulin, amino acids, and leucine stimulated rpS6 phosphorylation, and the

  8. Biodegradation of Leonardite by an alkali-producing bacterial community and characterization of the degraded products.

    Science.gov (United States)

    Gao, Tong-Guo; Jiang, Feng; Yang, Jin-Shui; Li, Bao-Zhen; Yuan, Hong-Li

    2012-03-01

    In this study, three bacterial communities were obtained from 12 Leonardite samples with the aim of identifying a clean, effective, and economic technique for the dissolution of Leonardite, a type of low-grade coal, in the production of humic acid (HA). The biodegradation ability and characteristics of the degraded products of the most effective bacterial community (MCSL-2), which degraded 50% of the Leonardite within 21 days, were further investigated. Analyses of elemental composition, (13)C NMR, and Fourier transform infrared revealed that the contents of C, O, and aliphatic carbon were similar in biodegraded humic acid (bHA) and chemically (alkali) extracted humic acid (cHA). However, the N and carboxyl carbon contents of bHA was higher than that of cHA. Furthermore, a positive correlation was identified between the degradation efficiency and the increasing pH of the culture medium, while increases of manganese peroxidase and esterase activities were also observed. These data demonstrated that both alkali production and enzyme reactions were involved in Leonardite solubilization by MCSL-2, although the former mechanism predominated. No fungus was observed by microscopy. Only four bacterial phylotypes were recognized, and Bacillus licheniformis-related bacteria were identified as the main group in MCSL-2 by analysis of amplified 16S rRNA genes, thus demonstrating that Leonardite degradation ability has a limited distribution in bacteria. Hormone-like bioactivities of bHA were also detected. In this study, a bacterial community capable of Leonardite degradation was identified and the products characterized. These data implicate the use of such bacteria for the exploitation of Leonardite as a biofertilizer.

  9. Impact of the NGF maturation and degradation pathway on the cortical cholinergic system phenotype.

    Science.gov (United States)

    Allard, Simon; Leon, Wanda C; Pakavathkumar, Prateep; Bruno, Martin A; Ribeiro-da-Silva, Alfredo; Cuello, A Claudio

    2012-02-08

    Cortical cholinergic atrophy plays a significant role in the cognitive loss seen with aging and in Alzheimer's disease (AD), but the mechanisms leading to it remain unresolved. Nerve growth factor (NGF) is the neurotrophin responsible for the phenotypic maintenance of basal forebrain cholinergic neurons in the mature and fully differentiated CNS. In consequence, its implication in cholinergic atrophy has been suspected; however, no mechanistic explanation has been provided. We have previously shown that the precursor of NGF (proNGF) is cleaved extracellularly by plasmin to form mature NGF (mNGF) and that mNGF is degraded by matrix metalloproteinase 9. Using cognitive-behavioral tests, Western blotting, and confocal and electron microscopy, this study demonstrates that a pharmacologically induced chronic failure in extracellular NGF maturation leads to a reduction in mNGF levels, proNGF accumulation, cholinergic degeneration, and cognitive impairment in rats. It also shows that inhibiting NGF degradation increases endogenous levels of the mature neurotrophin and increases the density of cortical cholinergic boutons. Together, the data point to a mechanism explaining cholinergic loss in neurodegenerative conditions such as AD and provide a potential therapeutic target for the protection or restoration of this CNS transmitter system in aging and AD.

  10. Bioremediation of soil polluted with crude oil and its derivatives: Microorganisms, degradation pathways, technologies

    Directory of Open Access Journals (Sweden)

    Beškoski Vladimir P.

    2012-01-01

    Full Text Available The contamination of soil and water with petroleum and its products occurs due to accidental spills during exploitation, transport, processing, storing and use. In order to control the environmental risks caused by petroleum products a variety of techniques based on physical, chemical and biological methods have been used. Biological methods are considered to have a comparative advantage as cost effective and environmentally friendly technologies. Bioremediation, defined as the use of biological systems to destroy and reduce the concentrations of hazardous waste from contaminated sites, is an evolving technology for the removal and degradation of petroleum hydrocarbons as well as industrial solvents, phenols and pesticides. Microorganisms are the main bioremediation agents due to their diverse metabolic capacities. In order to enhance the rate of pollutant degradation the technology optimizes the conditions for the growth of microorganisms present in soil by aeration, nutrient addition and, if necessary, by adding separately prepared microorganisms cultures. The other factors that influence the efficiency of process are temperature, humidity, presence of surfactants, soil pH, mineral composition, content of organic substance of soil as well as type and concentration of contaminant. This paper presents a review of our ex situ bioremediation procedures successfully implemented on the industrial level. This technology was used for treatment of soils contaminated by crude oil and its derivatives originated from refinery as well as soils polluted with oil fuel and transformer oil.

  11. Electrochemical degradation of sulfonamides at BDD electrode: kinetics, reaction pathway and eco-toxicity evaluation.

    Science.gov (United States)

    Fabiańska, Aleksandra; Białk-Bielińska, Anna; Stepnowski, Piotr; Stolte, Stefan; Siedlecka, Ewa Maria

    2014-09-15

    The investigation dealt with electrochemical oxidation of five sulfonamides (SNs): sulfadiazine (SDZ), sulfathiazole (STZ), sulfamerazine (SMR), sulfamethazine (SMN) and sulfadimethoxine (SDM) in aqueous solution at boron-doped diamond (BDD) electrode. All studied sulfonamides were degraded according to a pseudo first order kinetics. The structure of SNs had no significant effect on the values of pseudo first order rate constants. Increased degradation efficiency was observed in higher temperature and in acidic pH. Due to the presence of chlorine and nitrate SNs were more effectively oxidized from municipal wastewater treatment plant (WWTP) effluents than from pure supporting electrolyte Na2SO4. The intermediates identified by LC-MS and GC-MS analysis suggested that the hydroxyl radicals attack mainly the SN bond, but also the aromatic ring systems (aniline, pyrimidine or triazole) of SNs. Finally, the toxicity of the SNs solutions and effluents after electrochemical treatment was assessed through the measurement of growth inhibition of green algae (Scenedesmus vacualatus) and duckweed (Lemna minor). Toxicity of SMR, STZ, SMN solutions before and after electrochemical oxidation and SDM solution after the process in L. minor test was observed. No significant toxicity of studied SNs was observed in algae test. Copyright © 2014. Published by Elsevier B.V.

  12. Forced degradation of fingolimod: effect of co-solvent and characterization of degradation products by UHPLC-Q-TOF-MS/MS and 1H NMR.

    Science.gov (United States)

    Patel, Prinesh N; Kalariya, Pradipbhai D; Gananadhamu, S; Srinivas, R

    2015-11-10

    Fingolimod (FGL), an immunomodulator drug for treating multiple sclerosis, was subjected to hydrolysis (acidic, alkaline and neutral), oxidation, photolysis and thermal stress, as per International Conference on Harmonization specified conditions. The drug showed extensive degradation under base hydrolysis, however, it was stable under all other conditions. A total of three degradation products (DPs) were observed. The chromatographic separation of the drug and its degradation products was achieved on a Fortis C18 (100×2.1mm, 1.7μm) column with a mobile phase composed of 0.1% formic acid (Solvent A) and acetonitrile (Solvent B) in gradient mode. All the DPs were identified and characterized by liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) in combination with accurate mass measurements. The major DP was isolated and characterized by Nuclear Magnetic resonance spectroscopy. This is a typical case of degradation where acetonitrile used as co-solvent in stress studies, reacts with FGL in base hydrolytic conditions to produce acetylated DPs. Hence, it can be suggested that acetonitrile is not preferable as a co-solvent for stress degradation of FGL. The developed UHPLC method was validated as per ICH guidelines.

  13. The potential of photoacoustic microscopy as a tool to characterize the in vivo degradation of surgical sutures.

    Science.gov (United States)

    Aguirre, Juan; Morales-Dalmau, Jordi; Funk, Lutz; Jara, Francesc; Turon, Pau; Durduran, Turgut

    2014-08-01

    The ex vivo and in vivo imaging, and quantitative characterization of the degradation of surgical sutures (∼500 μm diameter) up to ∼1cm depth is demonstrated using a custom dark-field photo-acoustic microscope (PAM). A practical algorithm is developed to accurately measure the suture diameter during the degradation process. The results from tissue simulating phantoms and mice are compared to ex vivo measurements with an optical microscope demonstrating that PAM has a great deal of potential to characterize the degradation process of surgical sutures. The implications of this work for industrial applications are discussed.

  14. Time evolution and competing pathways in photodegradation of trifluralin and three of its major degradation products.

    Science.gov (United States)

    Tagle, Martín G Sarmiento; Laura Salum, María; Buján, Elba I; Argüello, Gustavo A

    2005-11-01

    The herbicide trifluralin (I)(N,N-di-n-propyl-2,6-dinitro-4-trifluoromethylaniline) decomposes, by the action of UV-Vis light (lambda > 300 nm), to several products, the most important (because they give subsequent photochemical reactions) being N-n-propyl-2,6-dinitro-4-trifluoromethylaniline (VI), 2-ethyl-7-nitro-5-trifluoromethyl-1H-benzimidazole 3-oxide (VII) and 2,6-dinitro-4-trifluoromethylaniline (XII). The time evolution of degradation of trifluralin (I) and the aforementioned three main photoproducts was studied in water and acetonitrile as solvents. The pseudo-first order rate constants allow one to calculate the branching ratios for some of the reactions involved. The preference for either N-dealkylation or cyclization depends on the solvent employed. Dissolved oxygen accelerates the photodegradation, especially the dealkylation.

  15. Characterization of degradation products of idarubicin through LC-UV, MS(n) and LC-MS-TOF studies.

    Science.gov (United States)

    Kaushik, Dheeraj; Bansal, Gulshan

    2013-11-01

    Idarubicin was subjected to forced degradation under the ICH recommended conditions of hydrolysis, oxidation, dry heat and photolysis to characterize its possible impurities and/or degradation products. The drug was found unstable to acid hydrolysis at 85°C and to alkaline hydrolysis, and oxidation at room temperature. The hydrolytic and oxidative degradation products were resolved with gradient and isocratic elution, respectively on an Inertsil RP18 (250 mm × 4.6mm; 5 μ) column with HCOONH4 (20mM, pH 3.0) and acetonitrile. The drug degraded to two products (O-I and O-II) in oxidative condition, two products (K-I and K-II) in alkaline hydrolytic, and one product (A-I) in acidic hydrolytic conditions. The purity of each in the LC-UV chromatogram was ascertained through LC-PDA analysis. The products were characterized through +ESI-MS(n) studies on the drug and LC-MS-TOF studies on the degraded drug solutions. Based on the multistage mass fragmentation pattern of idarubicin and accurate mass analysis of the degradation products, the O-I, O-II and A-I were characterized as desacetylidarubicin hydroperoxide, desacetylidarubicin and deglucosaminylidarubicin, respectively. The products K-I and K-II were not characterized due to their low concentrations and/or extremely weak ionization. The mechanisms of degradation of idarubicin to these products were proposed and discussed.

  16. Trafficking and degradation pathways in pathogenic conversion of prions and prion-like proteins in neurodegenerative diseases.

    Science.gov (United States)

    Victoria, Guiliana Soraya; Zurzolo, Chiara

    2015-09-02

    Several neurodegenerative diseases such as transmissible spongiform encephalopathies, Alzheimer's and Parkinson's diseases are caused by the conversion of cellular proteins to a pathogenic conformer. Despite differences in the primary structure and subcellular localization of these proteins, which include the prion protein, α-synuclein and amyloid precursor protein (APP), striking similarity has been observed in their ability to seed and convert naïve protein molecules as well as transfer between cells. This review aims to cover what is known about the intracellular trafficking of these proteins as well as their degradation mechanisms and highlight similarities in their movement through the endocytic pathway that could contribute to the pathogenic conversion and seeding of these proteins which underlies the basis of these diseases.

  17. Lipid rafts participate in aberrant degradative autophagic-lysosomal pathway of amyloid-beta peptide in Alzheimer’s disease

    Institute of Scientific and Technical Information of China (English)

    Xin Zhou; Chun Yang; Yufeng Liu; Peng Li; Huiying Yang; Jingxing Dai; Rongmei Qu; Lin Yuan

    2014-01-01

    Amyloid-beta peptide is the main component of amyloid plaques, which are found in Alzhei-mer’s disease. The generation and deposition of amyloid-beta is one of the crucial factors for the onset and progression of Alzheimer’s disease. Lipid rafts are glycolipid-rich liquid domains of the plasma membrane, where certain types of protein tend to aggregate and intercalate. Lipid rafts are involved in the generation of amyloid-beta oligomers and the formation of amyloid-beta peptides. In this paper, we review the mechanism by which lipid rafts disturb the aberrant deg-radative autophagic-lysosomal pathway of amyloid-beta, which plays an important role in the pathological process of Alzheimer’s disease. Moreover, we describe this mechanism from the view of the Two-system Theory of fasciology and thus, suggest that lipid rafts may be a new target of Alzheimer’s disease treatment.

  18. Synthesis, Characterization and in vitro Degradation Properties of Poly (Propylene Fumarate-co-Propylene Sebacate) Networks

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Poly(propylene fumarate-co-propylene sebacate) (P(PF-co-PS)) was crosslinked with N-vinyl pyrrolidone (N-VP) to form networks. It was investigated as biodegradable bone cement. In this paper, P(PF-co-PS) was synthesized and characterized by 1 H-NMR, FTIR and GPC. The effects of the amount of sebacate segments in P(PF-co-PS) main chains and the quantity of N-vinyl pyrrolidone on the in vitro degradation of the polymer networks were examined. Cylindrical specimens were submerged in phosphate buffered saline (PBS) at 37 ℃ and the pH value of PBS is 7.4 for 10 weeks. The gravimetry and compressive mechanical properties were tested over the degradation period. Networks formed by P(PF-co-PS)8020/N-VP exhibited higher weight loss and better mechanical properties when compared with poly(propylene fumarate)/N-VP networks. The mechanical properties of P(PF-co-PS)/N-VP can be maintained for a very long time, even for 70 days, the yield strength, fracture strength and compressive modulus are (51.78 ± 2.01) MPa, (52.331 ± 1.84)MPa and (957.78 ± 24.40) MPa, respectively. The results demonstrate that the compressive mechanical properties and degradation velocity can be modulated by the amount of crosslinking agents and sebacate segments along the main chains of copolymers.

  19. Biochemical Characterization of 3-Methyl-4-nitrophenol degradation in Burkholderia sp. Strain SJ98

    Directory of Open Access Journals (Sweden)

    Jun eMin

    2016-05-01

    Full Text Available Several strains have been reported to grow on 3-methyl-4-nitrophenol (3M4NP, the primary breakdown product of the excessively used insecticide fenitrothion. However, the microbial degradation of 3M4NP at molecular and biochemical levels remains unknown. Here, methyl-1,4-benzoquinone (MBQ and methylhydroquinone (MHQ,rather than catechol proposed previously, were identified as the intermediates before ring cleavage during 3M4NP degradation by Burkholderia sp. strain SJ98. Real-time quantitative PCR analysis indicated that the pnpABA1CDEF cluster involved in para-nitrophenol (PNP and 2-chloro-4-nitrophenol (2C4NP catabolism was also likely responsible for 3M4NP degradation in this strain. Purified PNP 4-monooxygenase (PnpA is able to catalyze the monooxygenation of 3M4NP to MBQ and exhibited an apparent Km value of 20.3±2.54 μM for 3M4NP, and pnpA is absolutely necessary for the catabolism of 3M4NP by gene knock-out and complementation. PnpB, a 1,4-benzoquinone reductase catalyzes the reduction of MBQ to MHQ, and also found to enhance PnpA activity in vitro in the conversion of 3M4NP to MBQ. By sequential catalysis assays, PnpCD, PnpE and PnpF were likely involved in the lower pathway of 3M4NP catabolism. Although NpcCD, NpcE and NpcF are able to catalyze the sequential conversion of MHQ in vitro, these enzymes are unlikely involved in 3M4NP catabolism because their coding genes were not upregulated by 3M4NP induction in vivo. These results revealed that the enzymes involved in PNP and 2C4NP catabolism were also responsible for 3M4NP degradation in strain SJ98. This fills a gap in our understanding of the microbial degradation of 3M4NP at molecular and biochemical levels and also provides another example to illustrate the adaptive flexibility in microbial catabolism for structurally similar compounds.

  20. Key enzymes of the protocatechuate branch of the β-ketoadipate pathway for aromatic degradation in Corynebacterium glutamicum

    Institute of Scientific and Technical Information of China (English)

    SHEN; Xihui; LIU; Shuangjiang

    2005-01-01

    Although the protocatechuate branch of the β-ketoadipate pathway in Gram bacteria has been well studied, this branch is less understood in Gram+ bacteria. In this study,Corynebacterium glutamicum was cultivated with protocatechuate, p-cresol, vanillate and 4-hydroxybenzoate as sole carbon and energy sources for growth. Enzymatic assays indicated that growing cells on these aromatic compounds exhibited protocatechuate 3,4-dioxygenase activities. Data-mining of the genome of this bacterium revealed that the genetic locus ncg12314-ncg12315 encoded a putative protocatechuate 3,4-dioxygenase. The genes,ncg12314 and ncg12315, were amplified by PCR technique and were cloned into plasmid (pET21aP34D). Recombinant Escherichia coli strain harboring this plasmid actively expressed protocatechuate 3,4-dioxygenase activity. Further, when this locus was disrupted in C. glutamicum, the ability to degrade and assimilate protocatechuate, p-cresol, vanillate or 4-hydroxybenzoate was lost and protocatechuate 3,4-dioxygenase activity was disappeared. The ability to grow with these aromatic compounds and protocatechuate 3,4-dioxygenase activity of C.glutamicum mutant could be restored by gene complementation. Thus, it is clear that the key enzyme for ring-cleavage, protocatechuate 3,4-dioxygenase, was encoded by ncg12314 and ncg12315. The additional genes involved in the protocatechuate branch of the β-ketoadipate pathway were identified by mining the genome data publically available in the GenBank. The functional identification of genes and their unique organization in C. glutamicum provided new insight into the genetic diversity of aromatic compound degradation.

  1. Degradation of pyrene and characterization of Saccharothrix sp. PYX-6 from the oligotrophic Tianchi Lake in Xinjiang Uygur Autonomous Region, China

    Institute of Scientific and Technical Information of China (English)

    HU Yuting; REN Fenghua; ZHOU Peijin; XIA Min; LIU Shuangjiang

    2003-01-01

    Bacterial strain PYX-6 that utilizes anthracene, phenanthrene, or pyrene for carbon and energy sources for growth was isolated from a non-polluted lake (Tianchi Lake) in Xinjiang Uygur Autonomons Region of China. Its morphology, physiological and biochemical properties, cell wall pattern and G+C mol% content of DNA molecules were characterized. The 16S rRNA gene of strain PYX-6 was sequenced and analyzed for similarities to related bacterial species. Results indicated that strain PYX-6 is a member of the Genus Saccharothrix, and the strain was named Saccharothrix sp. PYX-6. When pyrene was the sole carbon source in cultural medium, the strain PYX-6 assimilated pyrene for growth and 0.005% of yeast extract stimulated pyrene degradation and assimilation. The optimal pH of cultural medium and the optimal shaking frequency during cultivation were 6-8 and 200 r/m, respectively. It was found that the disappearance of pyrene in medium occurred before significant growth of strain PYX-6 took place. Phthalic acid, benzylacetic acid, and benzylpropenoic acid were detected as catabolic intermediates during pyrene degradation with mass spectroscopy and this result indicated that Saccharothrix sp. PYX-6 adopted a pathway that is different from the pathway of the previously reported pyrene-degrading Mycobacterium sp. PYR-1.

  2. Characterization of tobacco expressing functional oat phytochrome. Domains responsible for the rapid degradation of Pfr are conserved between monocots and dicots

    Energy Technology Data Exchange (ETDEWEB)

    Cherry, J.R.; Vierstra, R.D. (Univ. of Wisconsin, Madison (United States)); Hershey, H.P. (E.I.du Pont de Nemours and Co., Wilmington, DE (United States))

    1991-07-01

    Constitutive expression of a chimeric oat phytochrome gene in tobacco (Nicotiana tabacum) results in the accumulation of a functional 124-kilodalton photoreceptor that markedly alters the phenotype of light-grown tobacco. Here, we provide a detailed phenotypic and biochemical characterization of homozygous tobacco expressing high levels of oat phytochrome. Phenotypic changes include a substantial inhibition of stem elongation, decreased apical dominance, increased leaf chlorophyll content, and delayed leaf senescence. Oat phytochrome synthesized in tobacco is indistinguishable from that present in etiolated oats, having photoreversible difference spectrum maxima at 665 and 730 nanometers, exhibiting negligible dark reversion of phytochrome - far red-absorbing from (Pfr) to phytochrome - red-absorbing form (Pr), and existing as a dimer with an apparent size of approximately 300 kilodaltons. Heterodimers between the oat and tobacco chromoproteins were detected. Endogenous tobacco phytochrome and transgenically expressed oat phytochrome are rapidly degraded in vivo upon photoconversion of Pr to Pfr. Breakdown of both oat and tobacco Pfr is associated with the accumulation of ubiquitin-phytochrome conjugates, suggesting that degradation occurs via the ubiquitin-dependent proteolytic pathway. This result indicates that the factors responsible for selective recognition of Pfr by the ubiquitin pathway are conserved between monocot and dicot phytochromes. More broadly, it demonstrates that the domains(s) within a plant protein responsible for its selective breakdown can be recognized by the degradation machinery of heterologous species.

  3. Oncogenic activation of the Met receptor tyrosine kinase fusion protein, Tpr-Met, involves exclusion from the endocytic degradative pathway.

    Science.gov (United States)

    Mak, H H L; Peschard, P; Lin, T; Naujokas, M A; Zuo, D; Park, M

    2007-11-01

    Multiple mechanisms of dysregulation of receptor tyrosine kinases (RTKs) are observed in human cancers. In addition to gain-of-function, loss of negative regulation also contributes to oncogenic activation of RTKs. Negative regulation of many RTKs involves their internalization and degradation in the lysosome, a process regulated through ubiquitination. RTK oncoproteins activated following chromosomal translocation, are no longer transmembrane proteins, and are predicted to escape lysosomal degradation. To test this, we used the Tpr-Met oncogene, generated following chromosomal translocation of the hepatocyte growth factor receptor (Met). Unlike Met, Tpr-Met is localized in the cytoplasm and also lacks the binding site for Cbl ubiquitin ligases. We determined whether subcellular localization of Tpr-Met, and/or loss of its Cbl-binding site, is important for oncogenic activity. Presence of a Cbl-binding site and ubiquitination of cytosolic Tpr-Met oncoproteins does not alter their transforming activity. In contrast, plasma membrane targeting allows Tpr-Met to enter the endocytic pathway, and Tpr-Met transforming activity as well as protein stability are decreased in a Cbl-dependent manner. We show that transformation by Tpr-Met is in part dependent on its ability to escape normal downregulatory mechanisms. This provides a paradigm for many RTK oncoproteins activated following chromosomal translocation.

  4. Biochemical, transcriptional and translational evidences of the phenol-meta-degradation pathway by the hyperthermophilic Sulfolobus solfataricus 98/2.

    Directory of Open Access Journals (Sweden)

    Alexia Comte

    Full Text Available Phenol is a widespread pollutant and a model molecule to study the biodegradation of monoaromatic compounds. After a first oxidation step leading to catechol in mesophilic and thermophilic microorganisms, two main routes have been identified depending on the cleavage of the aromatic ring: ortho involving a catechol 1,2 dioxygenase (C12D and meta involving a catechol 2,3 dioxygenase (C23D. Our work aimed at elucidating the phenol-degradation pathway in the hyperthermophilic archaea Sulfolobus solfataricus 98/2. For this purpose, the strain was cultivated in a fermentor under different substrate and oxygenation conditions. Indeed, reducing dissolved-oxygen concentration allowed slowing down phenol catabolism (specific growth and phenol-consumption rates dropped 55% and 39%, respectively and thus, evidencing intermediate accumulations in the broth. HPLC/Diode Array Detector and LC-MS analyses on culture samples at low dissolved-oxygen concentration (DOC  =  0.06 mg x L(-1 suggested, apart for catechol, the presence of 2-hydroxymuconic acid, 4-oxalocrotonate and 4-hydroxy-2-oxovalerate, three intermediates of the meta route. RT-PCR analysis on oxygenase-coding genes of S. solfataricus 98/2 showed that the gene coding for the C23D was expressed only on phenol. In 2D-DIGE/MALDI-TOF analysis, the C23D was found and identified only on phenol. This set of results allowed us concluding that S. solfataricus 98/2 degrade phenol through the meta route.

  5. Relative Contribution of Prolyl Hydroxylase-Dependent and -Independent Degradation of HIF-1alpha by Proteasomal Pathways in Cerebral Ischemia

    Directory of Open Access Journals (Sweden)

    Yomna Badawi

    2017-05-01

    Full Text Available Hypoxia inducible factor-1 (HIF-1 is a key regulator in hypoxia and can determine the fate of brain cells during ischemia. However, the mechanism of HIF-1 regulation is still not fully understood in ischemic brains. We tested a hypothesis that both the 26S and the 20S proteasomal pathways were involved in HIF-1α degradation under ischemic conditions. Using in vitro ischemic model (oxygen and glucose deprivation and a mouse model of middle cerebral artery occlusion, we tested effects of inhibitors of proteasomes and prolyl hydroxylase (PHD on HIF-1α stability and brain injury in cerebral ischemia. We observed that 30 and 60 min of oxygen-glucose deprivation significantly increased the 20S proteasomal activity. We demonstrated that proteasome inhibitors increased HIF-1α stabilization and cell viability and were more effective than PHD inhibitors in primary cultured cortical neurons exposed to oxygen and glucose deprivation. Furthermore, the administration of the proteasome inhibitor, epoxomicin, to mice resulted in smaller infarct size and brain edema than a PHD inhibitor. Our results indicate that 20S proteasomes are involved in HIF-1α degradation in ischemic neurons and that proteasomal inhibition provides more HIF-1α stabilization and neuroprotection than PHD inhibition in cerebral ischemia.

  6. Sources and Input Pathways of Glyphosate and its Degradation Product AMPA

    Science.gov (United States)

    Bischofberger, S.; Hanke, I.; Wittmer, I.; Singer, H.; Stamm, C.

    2009-04-01

    Despite being the pesticide used in the largest quantities worldwide, the environmental relevance of glyphosate has been considered low for many years. Reasons for this assessment were the observations that glyphosate degrades quickly into its degradation product AMPA and that it sorbs strongly to soil particles. Hence, little losses to water bodies had been expected. Research during the last few years however contradicts this expectation. Although glyphosate is a dominant pesticide used in agriculture, recent studies on other pesticides revealed that urban sources may play a significant role for water quality. Therefore this study compares glyphosate input into streams from agricultural and urban sources. For that purpose, a catchment of an area of 25 km2 was selected. It has by about 12'000 inhabitants and about 15 % of the area is used as arable land. Four sampling sites were selected in the river system in order to reflect different urban and agricultural sources. Additionally, we sampled a combined sewer overflow, a rain sewer and the outflow of a waste water treatment plant. At each site discharge was measured continuously from March to November 2007. During 16 rain events samples were taken by automatic devices at a high temporal resolution. To analyze the concentration of glyphosate and its degradation product AMPA, the samples were derivatized with FMOC-Cl at low pH conditions and then filtrated. The solid phase extraction was conducted with Strata-X sorbent cartridge. Glyphosate and AMPA were detected with API 4000 after the chromatography with X bridge column C18. To assure the data quality, interne standards of Glyphosate and AMPA were added to every sample. The limit of detection and quantification for glyphosate and AMPA are bellow 1ng/l. We analyzed two rain events at a high resolution for all stations and several events at the outlet of the catchment. We measured high glyphosate concentration in urban and agriculture dominated catchments with up to

  7. Characterization of Organic Solar Cell Devices and their Interfaces under Degradation: Imaging, Electrical and Mechanical Methods

    DEFF Research Database (Denmark)

    Corazza, Michael

    techniques were also employed in order to study the effect of degradation on the device structure and its interfaces. This was done by exploiting different techniques that measured different properties of the device: mechanical, imaging, and electrical. Mechanical characterization of roll-to-roll processed....... Finally, imaging of cross sections of an ITO-free roll-to-roll processed device was performed successfully using transmission electron microscopy. The cross sections were prepared both with focused-ion-beam and ultramicrotomy, which gave the possibility for effectively comparing these two techniques...... energy is one of the answers for renewable energy. In this thesis, the research has been conducted on polymer solar cells. In particular, the thesis deals with the extensive study of device lifetime, characterized with several methods: from bare benchmarking of the lifetimes, to more advanced...

  8. Preparation, characterization, and microbial degradation of specifically radiolabeled [C]lignocelluloses from marine and freshwater macrophytes.

    Science.gov (United States)

    Benner, R; Maccubbin, A E; Hodson, R E

    1984-02-01

    Specifically radiolabeled [C-lignin]lignocelluloses were prepared from the aquatic macrophytes Spartina alterniflora, Juncus roemerianus, Rhizophora mangle, and Carex walteriana by using [C]phenylalanine, [C]tyrosine, and [C]cinnamic acid as precursors. Specifically radiolabeled [C-polysaccharide]lignocelluloses were prepared by using [C]glucose as precursor. The rates of microbial degradation varied among [C-lignin]lignocelluloses labeled with different lignin precursors within the same plant species. To determine the causes of these differential rates, [C-lignin]lignocelluloses were thoroughly characterized for the distribution of radioactivity in nonlignin contaminants and within the lignin macromolecule. In herbaceous plants, significant amounts (8 to 24%) of radioactivity from [C]phenylalanine and [C]tyrosine were found associated with protein, although very little (3%) radioactivity from [C]cinnamic acid was associated with protein. Microbial degradation of radiolabeled protein resulted in overestimation of lignin degradation rates in lignocelluloses derived from herbaceous aquatic plants. Other differences in degradation rates among [C-lignin]lignocelluloses from the same plant species were attributable to differences in the amount of label being associated with ester-linked subunits of peripheral lignin. After acid hydrolysis of [C-polysaccharide]lignocelluloses, radioactivity was detected in several sugars, although most of the radioactivity was distributed between glucose and xylose. After 576 h of incubation with salt marsh sediments, 38% of the polysaccharide component and between 6 and 16% of the lignin component (depending on the precursor) of J. roemerianus lignocellulose was mineralized to CO(2); during the same incubation period, 30% of the polysaccharide component and between 12 and 18% of the lignin component of S. alterniflora lignocellulose was mineralized.

  9. Genetic Characterization of 2,4,6-Trichlorophenol Degradation in Cupriavidus necator JMP134▿

    Science.gov (United States)

    Sánchez, M. A.; González, B.

    2007-01-01

    The degradation pathway of 2,4,6-trichlorophenol (2,4,6-TCP), a hazardous pollutant, in the aerobic bacterium Cupriavidus necator JMP134(pJP4) (formerly Ralstonia eutropha JMP134) is encoded by the tcp genes. These genes are located in a genetic context, tcpRXABCYD, which resembles a putative catabolic operon. In this work, these gene sequences were individually disrupted and mutant strains were evaluated for their ability to grow on or degrade 2,4,6-TCP. The tcpX and tcpA mutants completely failed to degrade this compound. Although the tcpC mutant was also unable to grow on 2,4,6-TCP, it still transformed this chlorophenol to 6-chlorohydroquinol. In contrast, the tcpD mutant grew on 2,4,6-TCP, suggesting the presence of additional maleylacetate reductase-encoding genes. Five other open reading frames encoding maleylacetate reductases, in addition to the tcpD gene, were found in the genome of C. necator, and two of them provide this function in the tcpD mutant. The tcpR gene, encoding a putative LysR-type transcriptional regulator, was disrupted, and this mutant strain completely failed to grow on 2,4,6-TCP. Transcriptional fusion studies demonstrated that TcpR activates the expression of the tcp genes, responding specifically to 2,4,6-TCP. The transcriptional start of the tcp operon was mapped, and a putative σ70-type promoter was identified. PMID:17322325

  10. Degradation studies of pentoxifylline: Isolation and characterization of a novel gem-dihydroperoxide derivative as major oxidative degradation product.

    Science.gov (United States)

    Mone, Mahesh Kumar; Chandrasekhar, K B

    2010-11-02

    Pentoxifylline was subjected to various stress conditions and degradation profile was studied with conventional LCMS. Interestingly, under oxidative stress conditions the drug substance underwent distinct transformation to give rise to a single major degradation product. The structure of this product was elucidated using 1D, 2D NMR spectroscopy, high resolution mass spectrometry (Q-TOF LC/MS) and found to be a novel gem-dihydroperoxide, namely 1-(5,5-Bis-hydroperoxy-hexyl)-3,7-dimethyl-3,7-dihydro-purine-2,6-dione. An efficient stability indicating liquid chromatographic separation method was developed for pentoxifylline and its three degradation products (including two from base hydrolysis) using 1.8 microm, C18 reverse phase column and UHPLC. Baseline separation was achieved with a run time of 4 min. The analytical assay method was validated with respect to system suitability, specificity, linearity, range, precision, accuracy and robustness.

  11. Synthesis, characterization and application of ZnO-Ag as a nanophotocatalyst for organic compounds degradation, mechanism and economic study.

    Science.gov (United States)

    Mohammadzadeh, S; Olya, M E; Arabi, A M; Shariati, A; Khosravi Nikou, M R

    2015-09-01

    The current work deals with ZnO-Ag nanocomposites (in the wide range of x in the Zn1-xO-Agx chemical composition) synthesized using microwave assisted solution combustion method. The structural, morphological and optical properties of the samples were characterized by XRD (X-ray diffraction), FTIR (Fourier transform infrared spectrometry), SEM (scanning electron microscopy technique), EDX (energy dispersive X-ray spectrum), ICP (inductively coupled plasma technique), TEM (transmission electron microscopy), BET (Brunauer-Emmett-Teller method), UV-Vis (ultraviolet-visible spectrophotometer) and photoluminescence spectrophotometer. The photocatalytic activity of the ZnO-Ag was investigated by photo-degradation of Acid Blue 113 (AB 113) under UV illumination in a semi-batch reactor. This experiment showed that ZnO-Ag has much more excellent photocatalytic properties than ZnO synthesized by the same method. The enhanced photocatalytic activity was due to the decrease in recombination of photogenerated electron-holes. The results showed the improvement of ZnO photocatalytic activity and there is an optimum amount of Ag (3.5mol%) that needs to be doped with ZnO. The effect of operating parameters such as pH, catalyst dose and dye concentration were investigated. The reaction byproducts were identified by LC/MS (liquid chromatography/mass spectrometry) analysis and a pathway was proposed as well. Kinetic studies indicated that the decolorization process follows the first order kinetics. Also, the degradation percentage of AB 113 was determined using a total organic carbon (TOC) analyzer. Additionally, cost analysis of the process, the mechanism and the role of Ag were discussed.

  12. Abatement and degradation pathways of toluene in indoor air by positive corona discharge.

    Science.gov (United States)

    Van Durme, J; Dewulf, J; Sysmans, W; Leys, C; Van Langenhove, H

    2007-08-01

    Indoor air concentrations of volatile organic compounds often exceed outdoor levels by a factor of 5. There is much interest in developing new technologies in order to improve indoor air quality. In this work non-thermal plasma (DC positive corona discharge) is explored as an innovative technology for indoor air purification. An inlet gas stream of 10 l min(-1) containing 0.50+/-0.02 ppm toluene was treated by the plasma reactor in atmospheric conditions. Toluene removal proved to be achievable with a characteristic energy density epsilon(0) of 50 J l(-1). Removal efficiencies were higher for 26% relative humidity (epsilon(0)=35 J l(-1)), compared with those at increased humidities (50% relative humidity, epsilon(0)=49 J l(-1)). Reaction products such as formic acid, benzaldehyde, benzyl alcohol, 3-methyl-4-nitrophenol, 4-methyl-2-nitrophenol, 4-methyl-2-propyl furan, 5-methyl-2-nitrophenol, 4-nitrophenol, 2-methyl-4,6-dinitrophenol are identified by means of mass spectrometry. Based on these by-products a toluene degradation mechanism is proposed.

  13. Microbial oil-degradation under mild hydrostatic pressure (10 MPa): which pathways are impacted in piezosensitive hydrocarbonoclastic bacteria?

    Science.gov (United States)

    Scoma, Alberto; Barbato, Marta; Hernandez-Sanabria, Emma; Mapelli, Francesca; Daffonchio, Daniele; Borin, Sara; Boon, Nico

    2016-03-01

    Oil spills represent an overwhelming carbon input to the marine environment that immediately impacts the sea surface ecosystem. Microbial communities degrading the oil fraction that eventually sinks to the seafloor must also deal with hydrostatic pressure, which linearly increases with depth. Piezosensitive hydrocarbonoclastic bacteria are ideal candidates to elucidate impaired pathways following oil spills at low depth. In the present paper, we tested two strains of the ubiquitous Alcanivorax genus, namely A. jadensis KS_339 and A. dieselolei KS_293, which is known to rapidly grow after oil spills. Strains were subjected to atmospheric and mild pressure (0.1, 5 and 10 MPa, corresponding to a depth of 0, 500 and 1000 m, respectively) providing n-dodecane as sole carbon source. Pressures equal to 5 and 10 MPa significantly lowered growth yields of both strains. However, in strain KS_293 grown at 10 MPa CO2 production per cell was not affected, cell integrity was preserved and PO43- uptake increased. Analysis of its transcriptome revealed that 95% of its genes were downregulated. Increased transcription involved protein synthesis, energy generation and respiration pathways. Interplay between these factors may play a key role in shaping the structure of microbial communities developed after oil spills at low depth and limit their bioremediation potential.

  14. Microbial oil-degradation under mild hydrostatic pressure (10 MPa): which pathways are impacted in piezosensitive hydrocarbonoclastic bacteria?

    KAUST Repository

    Scoma, Alberto

    2016-03-29

    Oil spills represent an overwhelming carbon input to the marine environment that immediately impacts the sea surface ecosystem. Microbial communities degrading the oil fraction that eventually sinks to the seafloor must also deal with hydrostatic pressure, which linearly increases with depth. Piezosensitive hydrocarbonoclastic bacteria are ideal candidates to elucidate impaired pathways following oil spills at low depth. In the present paper, we tested two strains of the ubiquitous Alcanivorax genus, namely A. jadensis KS_339 and A. dieselolei KS_293, which is known to rapidly grow after oil spills. Strains were subjected to atmospheric and mild pressure (0.1, 5 and 10 MPa, corresponding to a depth of 0, 500 and 1000 m, respectively) providing n-dodecane as sole carbon source. Pressures equal to 5 and 10 MPa significantly lowered growth yields of both strains. However, in strain KS_293 grown at 10 MPa CO2 production per cell was not affected, cell integrity was preserved and PO43− uptake increased. Analysis of its transcriptome revealed that 95% of its genes were downregulated. Increased transcription involved protein synthesis, energy generation and respiration pathways. Interplay between these factors may play a key role in shaping the structure of microbial communities developed after oil spills at low depth and limit their bioremediation potential.

  15. Copper-promoted circumneutral activation of H2O2 by magnetic CuFe2O4 spinel nanoparticles: Mechanism, stoichiometric efficiency, and pathway of degrading sulfanilamide.

    Science.gov (United States)

    Feng, Yong; Liao, Changzhong; Shih, Kaimin

    2016-07-01

    To evaluate the heterogeneous degradation of sulfanilamide by external energy-free Fenton-like reactions, magnetic CuFe2O4 spinel nanoparticles (NPs) were synthesized and used as catalysts for activation of hydrogen peroxide (H2O2). The physicochemical properties of the CuFe2O4 NPs were characterized with several techniques, including X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, and magnetometry. In the catalytic experiments, CuFe2O4 NPs/H2O2 oxidation showed the best degradation performance in the circumneutral conditions that resulted from the presence of Cu(II) on the surface of the CuFe2O4 NPs. The surface area-normalized pseudo-first-order rate constants were calculated as 2.60 × 10(-2) L m(-1) min(-1), 2.58 × 10(-3) L m(-1) min(-1), 1.92 × 10(-3) L m(-1) min(-1), and 7.30 × 10(-4) L m(-1) min(-1) for CuO, CuFe2O4 NPs, Fe3O4, and α-Fe2O3 catalysts, respectively. Thus, solid state Cu(II) was more reactive and efficient than Fe(III) in the circumneutral activation of H2O2; this finding was further supported by the results regarding the stoichiometric efficiency of H2O2. The effects of experimental parameters such as the oxidant dosage and catalyst loading were investigated. The mechanism for H2O2 activation on the spinel surface was explored and could be explained by the solid redox cycles of Fe(II)/Fe(III) and Cu(II)/Cu(I). Based on the products detected, a degradation pathway via the CS bond cleavage is proposed for the degradation of sulfanilamide. The findings of this study suggest that copper can be used as a doping metal to improve the reactivity and expand the effective pH range of iron oxides. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Genome-wide investigation and functional characterization of the β-ketoadipate pathway in the nitrogen-fixing and root-associated bacterium Pseudomonas stutzeri A1501

    Directory of Open Access Journals (Sweden)

    Geng Lizhao

    2010-02-01

    Full Text Available Abstract Background Soil microorganisms are mainly responsible for the complete mineralization of aromatic compounds that usually originate from plant products or environmental pollutants. In many cases, structurally diverse aromatic compounds can be converted to a small number of structurally simpler intermediates, which are metabolized to tricarboxylic acid intermediates via the β-ketoadipate pathway. This strategy provides great metabolic flexibility and contributes to increased adaptation of bacteria to their environment. However, little is known about the evolution and regulation of the β-ketoadipate pathway in root-associated diazotrophs. Results In this report, we performed a genome-wide analysis of the benzoate and 4-hydroxybenzoate catabolic pathways of Pseudomonas stutzeri A1501, with a focus on the functional characterization of the β-ketoadipate pathway. The P. stutzeri A1501 genome contains sets of catabolic genes involved in the peripheral pathways for catabolism of benzoate (ben and 4-hydroxybenzoate (pob, and in the catechol (cat and protocatechuate (pca branches of the β-ketoadipate pathway. A particular feature of the catabolic gene organization in A1501 is the absence of the catR and pcaK genes encoding a LysR family regulator and 4-hydroxybenzoate permease, respectively. Furthermore, the BenR protein functions as a transcriptional activator of the ben operon, while transcription from the catBC promoter can be activated in response to benzoate. Benzoate degradation is subject to carbon catabolite repression induced by glucose and acetate in A1501. The HPLC analysis of intracellular metabolites indicated that low concentrations of 4-hydroxybenzoate significantly enhance the ability of A1501 to degrade benzoate. Conclusions The expression of genes encoding proteins involved in the β-ketoadipate pathway is tightly modulated by both pathway-specific and catabolite repression controls in A1501. This strain provides an ideal

  17. Urothelial endocytic vesicle recycling and lysosomal degradative pathway regulated by lipid membrane composition.

    Science.gov (United States)

    Grasso, E J; Calderón, R O

    2013-02-01

    The urothelium, a specialized epithelium that covers the mucosa cell surface of the urinary bladder, undergoes dramatic morphological changes during the micturition cycle that involve a membrane apical traffic. This traffic was first described as a lysosomal pathway, in addition to the known endocytosis/exocytosis membrane recycling. In an attempt to understand the role of membrane lipid composition in those effects, we previously described the lipid-dependent leakage of the endocytosed vesicle content. In this work, we demonstrated clear differences in the traffic of both the fluid probe and the membrane-bound probe in urothelial umbrella cells by using spectrofluorometry and/or confocal and epifluorescence microscopy. Different membrane lipid compositions were established by using three diet formulae enriched in oleic acid, linoleic acid and a commercial formula. Between three and five animals for each dietary treatment were used for each analysis. The decreased endocytosis of both fluid and membrane-bound probes (approximately 32 and 49 % lower, respectively) in oleic acid-derived umbrella cells was concomitant with an increased recycling (approximately 4.0 and 3.7 times, respectively) and diminished sorting to the lysosome (approximately 23 and 37 %, respectively) when compared with the control umbrella cells. The higher intravesicular pH and the impairment of the lysosomal pathway of oleic acid diet-derived vesicles compared to linoleic acid diet-derived vesicles and control diet-derived vesicles correlate with our findings of a lower V-ATPase activity previously reported. We integrated the results obtained in the present and previous work to determine the sorting of endocytosed material (fluid and membrane-bound probes) into the different cell compartments. Finally, the weighted average effect of the individual alterations on the intracellular distribution was evaluated. The results shown in this work add evidences for the modulatory role of the membrane

  18. Potentiality of yeast Candida sp. SMN04 for degradation of cefdinir, a cephalosporin antibiotic: kinetics, enzyme analysis and biodegradation pathway.

    Science.gov (United States)

    Selvi, A; Das, Devlina; Das, Nilanjana

    2015-01-01

    A new yeast strain isolated from the pharmaceutical wastewater was capable of utilizing cefdinir as a sole carbon source for their growth in mineral medium. The yeast was identified and named as Candida sp. SMN04 based on morphology and 18S-ITS-D1/D2/D3 rRNA sequence analysis. The interaction between factors pH (3.0-9.0), inoculum dosage (1-7%), time (1-11 day) and cefdinir concentration (50-450 mg/L) was studied using a Box-Behnken design. The factors were studied as a result of their effect on cell dry weight (R1; g/L), extended spectrum β-lactamase (ESBL) assay (R2; mm), P450 activity (R3; U/mL) and degradation (R4; %). Maximum values of R1, R2, R3 and R4 were obtained at central values of all the parameters. The isolated yeast strain efficiently degraded 84% of 250 mg L⁻¹ of cefdinir within 6 days with a half-life of 2.97 days and degradation rate constant of 0.2335 per day. Pseudo-first-order model efficiently described the process. Among the various enzymes tested, the order of activity at the end of Day 4 was noted to be: cytochrome P450 (1.76 ± 0.03) > NADPH reductase (1.51 ± 0.20) > manganese peroxidase and amylase (0.66 ± 0.15; 0.66 ± 0.70). Intermediates were successfully characterized by liquid chromatography-mass spectrometry. The opening of the β-lactam ring involving ESBL activity was considered as one of the major steps in the cefdinir degradation process. Fourier transform-infrared spectroscopy analysis showed the absence of spectral vibrations between 1766 and 1519 cm⁻¹ confirming the complete removal of lactam ring during cefdinir degradation. The results of the present study are promising for the use of isolated yeast Candida sp. SMN04 as a potential bioremediation agent.

  19. Purification and characterization of the enzymes involved in nicotinamide adenine dinucleotide degradation by Penicillium brevicompactum NRC 829.

    Science.gov (United States)

    Ali, Thanaa Hamed; El-Ghonemy, Dina Helmy

    2016-06-01

    The present study was conducted to investigate a new pathway for the degradation of nicotinamide adenine dinucleotide (NAD) by Penicillium brevicompactum NRC 829 extracts. Enzymes involved in the hydrolysis of NAD, i.e. alkaline phosphatase, aminohydrolase and glycohydrolase were determined. Alkaline phosphatase was found to catalyse the sequential hydrolysis of two phosphate moieties of NAD molecule to nicotinamide riboside plus adenosine. Adenosine was then deaminated by aminohydrolase to inosine and ammonia. While glycohydrolase catalyzed the hydrolysis of the nicotinamide-ribosidic bond of NAD+ to produce nicotinamide and ADP-ribose in equimolar amounts, enzyme purification through a 3-step purification procedure revealed the existence of two peaks of alkaline phosphatases, and one peak contained deaminase and glycohydrolase activities. NAD deaminase was purified to homogeneity as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis with an apparent molecular mass of 91 kDa. Characterization and determination of some of NAD aminohydrolase kinetic properties were conducted due to its biological role in the regulation of cellular NAD level. The results also revealed that NAD did not exert its feedback control on nicotinamide amidase produced by P. brevicompactum.

  20. Cx31 is assembled and trafficked to cell surface by ER-Golgi pathway and degraded by proteasomal or lysosomal pathways

    Institute of Scientific and Technical Information of China (English)

    Li Qiang HE; Zhi Gao LONG; He Ping DAI; Kun XIA; Jia Hui XIA; Zhuo Hua ZHANG; Fang CAI; Yu LIU; Mu Jun LIU; Zhi Ping TAN; Qian PAN; Fai Yan FANG; De Sheng LIANG; Ling Qian WU

    2005-01-01

    Gap junctions, consisting of connexins, allow the exchange of small molecules (<1 kD) between adjacent cells, thus providing a mechanism for synchronizing the responses of groups of cells to environmental stimuli. Connexin 31 is a member of the connexin family. Mutations on connexin 31 are associated with erythrokeratodermia variabilis, hearing impairment and peripheral neuropathy. However, the pathological mechanism for connexin 31 mutants in these diseases are still unknown. In this study, we analyzed the assembly, trafficking and metabolism of connexin 31 in HeLa cells stably expressing connexin 31. Calcein transfer assay showed that calcein transfer was inhibited when cells were treated with Brefeldin A or cytochalasin D, but not when treated with nocodazole or α-glycyrrhetinic acid, suggesting that Golgi apparatus and actin filaments, but not microtubules, are crucial to the trafficking and assembly of connexin 31, as well as the formation of gap junction intercellular communication by connexin 31. Additionally, α-glycyrrhetinic acid did not effectively inhibit gap junctional intercellular communication formed by connexin 31. Pulse-chase assay revealed that connexin 31 had a half-life of about 6 h. Moreover, Western blotting and fluorescent staining demonstrated that in HeLa cells stably expressing connexin 31, the amount of connexin 31 was significantly increased after these cells were treated with proteasomal or lysosomal inhibitors. These findings indicate that connexin 31 was rapidly renewed,and possibly degraded by both proteasomal and lysosomal pathways.

  1. Characterization of an antennal carboxylesterase from the pest moth Spodoptera littoralis degrading a host plant odorant.

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    Nicolas Durand

    Full Text Available BACKGROUND: Carboxyl/cholinesterases (CCEs are highly diversified in insects. These enzymes have a broad range of proposed functions, in neuro/developmental processes, dietary detoxification, insecticide resistance or hormone/pheromone degradation. As few functional data are available on purified or recombinant CCEs, the physiological role of most of these enzymes is unknown. Concerning their role in olfaction, only two CCEs able to metabolize sex pheromones have been functionally characterized in insects. These enzymes are only expressed in the male antennae, and secreted into the lumen of the pheromone-sensitive sensilla. CCEs able to hydrolyze other odorants than sex pheromones, such as plant volatiles, have not been identified. METHODOLOGY: In Spodoptera littoralis, a major crop pest, a diversity of antennal CCEs has been previously identified. We have employed here a combination of molecular biology, biochemistry and electrophysiology approaches to functionally characterize an intracellular CCE, SlCXE10, whose predominant expression in the olfactory sensilla suggested a role in olfaction. A recombinant protein was produced using the baculovirus system and we tested its catabolic properties towards a plant volatile and the sex pheromone components. CONCLUSION: We showed that SlCXE10 could efficiently hydrolyze a green leaf volatile and to a lesser extent the sex pheromone components. The transcript level in male antennae was also strongly induced by exposure to this plant odorant. In antennae, SlCXE10 expression was associated with sensilla responding to the sex pheromones and to plant odours. These results suggest that a CCE-based intracellular metabolism of odorants could occur in insect antennae, in addition to the extracellular metabolism occurring within the sensillar lumen. This is the first functional characterization of an Odorant-Degrading Enzyme active towards a host plant volatile.

  2. Characterizing Autism Spectrum Disorders by Key Biochemical Pathways

    Directory of Open Access Journals (Sweden)

    Megha eSubramanian

    2015-09-01

    Full Text Available The genetic and phenotypic heterogeneity of autism spectrum disorders (ASD presents a substantial challenge for diagnosis, classification, research, and treatment. Investigations into the underlying molecular etiology of ASD have often yielded mixed and at times opposing findings. Defining the molecular and biochemical underpinnings of heterogeneity in ASD is crucial to our understanding of the pathophysiological development of the disorder, and has the potential to assist in diagnosis and the rational design of clinical trials. In this review, we propose that genetically diverse forms of ASD may be usefully parsed into entities resulting from converse patterns of growth regulation at the molecular level, which lead to the correlates of general synaptic and neural overgrowth or undergrowth. Abnormal brain growth during development is a characteristic feature that has been observed both in children with autism and in mouse models of autism. We review evidence from syndromic and non-syndromic ASD to suggest that entities currently classified as autism may fundamentally differ by underlying pro- or anti-growth abnormalities in key biochemical pathways, giving rise to either excessive or reduced synaptic connectivity in affected brain regions. We posit that this classification strategy has the potential not only to aid research efforts, but also to ultimately facilitate early diagnosis and direct appropriate therapeutic interventions.

  3. Characterizing autism spectrum disorders by key biochemical pathways.

    Science.gov (United States)

    Subramanian, Megha; Timmerman, Christina K; Schwartz, Joshua L; Pham, Daniel L; Meffert, Mollie K

    2015-01-01

    The genetic and phenotypic heterogeneity of autism spectrum disorders (ASD) presents a substantial challenge for diagnosis, classification, research, and treatment. Investigations into the underlying molecular etiology of ASD have often yielded mixed and at times opposing findings. Defining the molecular and biochemical underpinnings of heterogeneity in ASD is crucial to our understanding of the pathophysiological development of the disorder, and has the potential to assist in diagnosis and the rational design of clinical trials. In this review, we propose that genetically diverse forms of ASD may be usefully parsed into entities resulting from converse patterns of growth regulation at the molecular level, which lead to the correlates of general synaptic and neural overgrowth or undergrowth. Abnormal brain growth during development is a characteristic feature that has been observed both in children with autism and in mouse models of autism. We review evidence from syndromic and non-syndromic ASD to suggest that entities currently classified as autism may fundamentally differ by underlying pro- or anti-growth abnormalities in key biochemical pathways, giving rise to either excessive or reduced synaptic connectivity in affected brain regions. We posit that this classification strategy has the potential not only to aid research efforts, but also to ultimately facilitate early diagnosis and direct appropriate therapeutic interventions.

  4. Streptococcus pyogenes malate degradation pathway links pH regulation and virulence.

    Science.gov (United States)

    Paluscio, Elyse; Caparon, Michael G

    2015-03-01

    The ability of Streptococcus pyogenes to infect different niches within its human host most likely relies on its ability to utilize alternative carbon sources. In examining this question, we discovered that all sequenced S. pyogenes strains possess the genes for the malic enzyme (ME) pathway, which allows malate to be used as a supplemental carbon source for growth. ME is comprised of four genes in two adjacent operons, with the regulatory two-component MaeKR required for expression of genes encoding a malate permease (maeP) and malic enzyme (maeE). Analysis of transcription indicated that expression of maeP and maeE is induced by both malate and low pH, and induction in response to both cues is dependent on the MaeK sensor kinase. Furthermore, both maePE and maeKR are repressed by glucose, which occurs via a CcpA-independent mechanism. Additionally, malate utilization requires the PTS transporter EI enzyme (PtsI), as a PtsI(-) mutant fails to express the ME genes and is unable to utilize malate. Virulence of selected ME mutants was assessed in a murine model of soft tissue infection. MaeP(-), MaeK(-), and MaeR(-) mutants were attenuated for virulence, whereas a MaeE(-) mutant showed enhanced virulence compared to that of the wild type. Taken together, these data show that ME contributes to S. pyogenes' carbon source repertory, that malate utilization is a highly regulated process, and that a single regulator controls ME expression in response to diverse signals. Furthermore, malate uptake and utilization contribute to the adaptive pH response, and ME can influence the outcome of infection.

  5. Degradation of {gamma}-HCH spiked soil using stabilized Pd/Fe{sup 0} bimetallic nanoparticles: Pathways, kinetics and effect of reaction conditions

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Ritu [Ecotoxicology Division, CSIR-Indian Institute of Toxicology Research, Post Box 80, Mahatma Gandhi Marg, Lucknow 226 001, UP (India); Department of Environmental Science, Babasaheb Bhimrao Ambedkar University, Raebareli Road, Lucknow 226 025, UP (India); Misra, Virendra, E-mail: virendra_misra2001@yahoo.co.in [Ecotoxicology Division, CSIR-Indian Institute of Toxicology Research, Post Box 80, Mahatma Gandhi Marg, Lucknow 226 001, UP (India); Mudiam, Mohana Krishna Reddy [Analytical Chemistry Division, CSIR-Indian Institute of Toxicology Research, Post Box 80, Mahatma Gandhi Marg, Lucknow 226 001, UP (India); Chauhan, Lalit Kumar Singh [Petroleum Toxicology Division, CSIR-Indian Institute of Toxicology Research, Post Box 80, Mahatma Gandhi Marg, Lucknow 226 001, UP (India); Singh, Rana Pratap [Department of Environmental Science, Babasaheb Bhimrao Ambedkar University, Raebareli Road, Lucknow 226 025, UP (India)

    2012-10-30

    Highlights: Black-Right-Pointing-Pointer This study explores the potential of CMC-Pd/nFe{sup 0} to degrade {gamma}-HCH in spiked soil. Black-Right-Pointing-Pointer Sorption-desorption characteristics and partitioning of {gamma}-HCH is investigated. Black-Right-Pointing-Pointer Three degradation pathways has been proposed and discussed. Black-Right-Pointing-Pointer {gamma}-HCH degradation mechanism and kinetics is elucidated. Black-Right-Pointing-Pointer Activation energy reveals that {gamma}-HCH degradation is a surface mediated reaction. - Abstract: This study investigates the degradation pathway of gamma-hexachlorocyclohexane ({gamma}-HCH) in spiked soil using carboxymethyl cellulose stabilized Pd/Fe{sup 0} bimetallic nanoparticles (CMC-Pd/nFe{sup 0}). GC-MS analysis of {gamma}-HCH degradation products showed the formation of pentachlorocyclohexene, tri- and di-chlorobenzene as intermediate products while benzene was formed as the most stable end product. On the basis of identified intermediates and final products, degradation pathway of {gamma}-HCH has been proposed. Batch studies showed complete {gamma}-HCH degradation at a loading of 0.20 g/L CMC-Pd/nFe{sup 0} within 6 h of incubation. The surface area normalized rate constant (k{sub SA}) was found to be 7.6 Multiplication-Sign 10{sup -2} L min{sup -1} m{sup -2}. CMC-Pd/nFe{sup 0} displayed {approx}7-fold greater efficiency for {gamma}-HCH degradation in comparison to Fe{sup 0} nanoparticles (nFe{sup 0}), synthesized without CMC and Pd. Further studies showed that increase in CMC-Pd/nFe{sup 0} loading and reaction temperature facilitates {gamma}-HCH degradation, whereas a declining trend in degradation was noticed with the increase in pH, initial {gamma}-HCH concentration and in the presence of cations. The data on activation energy (33.7 kJ/mol) suggests that {gamma}-HCH degradation is a surface mediated reaction. The significance of the study with respect to remediation of {gamma}-HCH contaminated soil using

  6. Development and validation of stability-indicating RP-HPLC method for determination of Olmesartan medoxomile in pharmaceutical dosage form and identification, characterization of alkaline degradation impurity of Olmesartan medoxomile drug substance as well as drug product

    Directory of Open Access Journals (Sweden)

    Jain P.S.

    2012-01-01

    Full Text Available Olmesartan Medoxomil (OLME belongs to a group of angiotensin II receptor blockers used as an antihypertensive agent and is currently being used for prevention of Hypertension. This paper describes the Validation and development of stability indicating RP-HPLC method for the determination of OLME in the presence of its degradation products generated from forced degradation study and characterization of degradation product (impurity. The assay involved gradient elution of OLME on An LC GC BDS C18 column (250 × 4.5mm, 5-μm particle size was employed for loading the sample. The mobile phase A consists of 7 ml Triethylamine in 1000 ml water (pH adjusted to 3.0 with orthophosphoric acid and B contains acetonitrile. Quantification was achieved with photodiode array detection at 257 nm. The chromatographic separation was obtained with a retention time of 6.72 min, and the method was linear in the range 50-150 µg/ml. The method was validated according to the ICH guidelines with respect to linearity, precision, accuracy, limit of detection (LOD, limit of quantification (LOQ, specificity and robustness. Impurity found in stressed and stability studies of Olmesartan Medoxomil in both drug substance and drug product are described. This degradation product is identified as 1-(biphenyl-4-ylmethyl-1H-imidazole-5-carboxylic acid. An Alkaline degradation pathway of Olmesartan medoxomil, for the formation of this degradation product, has been proposed and degradation product was characterized.

  7. Metagenomic Analysis of Hot Springs in Central India Reveals Hydrocarbon Degrading Thermophiles and Pathways Essential for Survival in Extreme Environments

    Science.gov (United States)

    Saxena, Rituja; Dhakan, Darshan B.; Mittal, Parul; Waiker, Prashant; Chowdhury, Anirban; Ghatak, Arundhuti; Sharma, Vineet K.

    2017-01-01

    Extreme ecosystems such as hot springs are of great interest as a source of novel extremophilic species, enzymes, metabolic functions for survival and biotechnological products. India harbors hundreds of hot springs, the majority of which are not yet explored and require comprehensive studies to unravel their unknown and untapped phylogenetic and functional diversity. The aim of this study was to perform a large-scale metagenomic analysis of three major hot springs located in central India namely, Badi Anhoni, Chhoti Anhoni, and Tattapani at two geographically distinct regions (Anhoni and Tattapani), to uncover the resident microbial community and their metabolic traits. Samples were collected from seven distinct sites of the three hot spring locations with temperature ranging from 43.5 to 98°C. The 16S rRNA gene amplicon sequencing of V3 hypervariable region and shotgun metagenome sequencing uncovered a unique taxonomic and metabolic diversity of the resident thermophilic microbial community in these hot springs. Genes associated with hydrocarbon degradation pathways, such as benzoate, xylene, toluene, and benzene were observed to be abundant in the Anhoni hot springs (43.5–55°C), dominated by Pseudomonas stutzeri and Acidovorax sp., suggesting the presence of chemoorganotrophic thermophilic community with the ability to utilize complex hydrocarbons as a source of energy. A high abundance of genes belonging to methane metabolism pathway was observed at Chhoti Anhoni hot spring, where methane is reported to constitute >80% of all the emitted gases, which was marked by the high abundance of Methylococcus capsulatus. The Tattapani hot spring, with a high-temperature range (61.5–98°C), displayed a lower microbial diversity and was primarily dominated by a nitrate-reducing archaeal species Pyrobaculum aerophilum. A higher abundance of cell metabolism pathways essential for the microbial survival in extreme conditions was observed at Tattapani. Taken together

  8. Aqueous photochemical degradation of hydroxylated PAHs: Kinetics, pathways, and multivariate effects of main water constituents

    Energy Technology Data Exchange (ETDEWEB)

    Ge, Linke; Na, Guangshui [Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023 (China); Chen, Chang-Er [Lancaster Environment Centre, Lancaster University, Lancaster LA1 4YQ (United Kingdom); Li, Jun [Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023 (China); College of Marine Science, Shanghai Ocean University, Shanghai 201306 (China); Ju, Maowei; Wang, Ying; Li, Kai [Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023 (China); Zhang, Peng, E-mail: pzhang@nmemc.org.cn [Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023 (China); Yao, Ziwei [Key Laboratory for Ecological Environment in Coastal Areas (SOA), National Marine Environmental Monitoring Center, Dalian 116023 (China)

    2016-03-15

    PAHs. • Hydroxylated PAHs intrinsically photodegrade fast in sunlit surface waters. • Reaction types and transformation pathways of 9-Hydroxyfluorene were clarified. • Photolysis kinetics was affected by multivariate effects of main water constituents. • The photomodified toxicity of 9-Hydroxyfluorene was examined using Vibrio fischeri.

  9. Isolation of Alcaligenes sp strain L6 at low oxygen concentrations and degradation of 3-chlorobenzoate via a pathway not involving (chloro)catechols

    NARCIS (Netherlands)

    Krooneman, J; Wieringa, EBA; Moore, ERB; Gerritse, J; Prins, RA; Gottschal, JC

    1996-01-01

    Isolations of 3-chlorobenzoate (3CBA)-degrading aerobic bacteria under reduced O-2, partial pressures yielded organisms which metabolized 3CBA via the gentisate or the protocatechuate pathway rather than via the catechol route. The 3CBA metabolism of one of these isolates, L6, which,vas identified a

  10. Characterization of Trapped Lignin-Degrading Microbes in Tropical Forest Soil

    Energy Technology Data Exchange (ETDEWEB)

    DeAngelis, Kristen; Allgaier, Martin; Chavarria, Yaucin; Fortney, Julian; Hugenholtz, Phillip; Simmons, Blake; Sublette, Kerry; Silver, Whendee; Hazen, Terry

    2011-07-14

    Lignin is often the most difficult portion of plant biomass to degrade, with fungi generally thought to dominate during late stage decomposition. Lignin in feedstock plant material represents a barrier to more efficient plant biomass conversion and can also hinder enzymatic access to cellulose, which is critical for biofuels production. Tropical rain forest soils in Puerto Rico are characterized by frequent anoxic conditions and fluctuating redox, suggesting the presence of lignin-degrading organisms and mechanisms that are different from known fungal decomposers and oxygen-dependent enzyme activities. We explored microbial lignin-degraders by burying bio-traps containing lignin-amended and unamended biosep beads in the soil for 1, 4, 13 and 30 weeks. At each time point, phenol oxidase and peroxidase enzyme activity was found to be elevated in the lignin-amended versus the unamended beads, while cellulolytic enzyme activities were significantly depressed in lignin-amended beads. Quantitative PCR of bacterial communities showed more bacterial colonization in the lignin-amended compared to the unamended beads after one and four weeks, suggesting that the lignin supported increased bacterial abundance. The microbial community was analyzed by small subunit 16S ribosomal RNA genes using microarray (PhyloChip) and by high-throughput amplicon pyrosequencing based on universal primers targeting bacterial, archaeal, and eukaryotic communities. Community trends were significantly affected by time and the presence of lignin on the beads. Lignin-amended beads have higher relative abundances of representatives from the phyla Actinobacteria, Firmicutes, Acidobacteria and Proteobacteria compared to unamended beads. This study suggests that in low and fluctuating redox soils, bacteria could play a role in anaerobic lignin decomposition.

  11. Characterization of trapped lignin-degrading microbes in tropical forest soil

    Energy Technology Data Exchange (ETDEWEB)

    DeAngelis, K.M.; Allgaier, M.; Chavarria, Y.; Fortney, J.L.; Hugenholz, P.; Simmons, B.; Sublette, K.; Silver, W.L.; Hazen, T.C.

    2011-03-01

    Lignin is often the most difficult portion of plant biomass to degrade, with fungi generally thought to dominate during late stage decomposition. Lignin in feedstock plant material represents a barrier to more efficient plant biomass conversion and can also hinder enzymatic access to cellulose, which is critical for biofuels production. Tropical rain forest soils in Puerto Rico are characterized by frequent anoxic conditions and fluctuating redox, suggesting the presence of lignin-degrading organisms and mechanisms that are different from known fungal decomposers and oxygen-dependent enzyme activities. We explored microbial lignin-degraders by burying bio-traps containing lignin-amended and unamended biosep beads in the soil for 1, 4, 13 and 30 weeks. At each time point, phenol oxidase and peroxidase enzyme activity was found to be elevated in the lignin-amended versus the unamended beads, while cellulolytic enzyme activities were significantly depressed in lignin-amended beads. Quantitative PCR of bacterial communities showed more bacterial colonization in the lignin-amended compared to the unamended beads after one and four weeks, suggesting that the lignin supported increased bacterial abundance. The microbial community was analyzed by small subunit 16S ribosomal RNA genes using microarray (PhyloChip) and by high-throughput amplicon pyrosequencing based on universal primers targeting bacterial, archaeal, and eukaryotic communities. Community trends were significantly affected by time and the presence of lignin on the beads. Lignin-amended beads have higher relative abundances of representatives from the phyla Actinobacteria, Firmicutes, Acidobacteria and Proteobacteria compared to unamended beads. This study suggests that in low and fluctuating redox soils, bacteria could play a role in anaerobic lignin decomposition.

  12. Modern mass spectrometry in the characterization and degradation of biodegradable polymers

    Energy Technology Data Exchange (ETDEWEB)

    Rizzarelli, Paola, E-mail: paola.rizzarelli@cnr.it; Carroccio, Sabrina

    2014-01-15

    Graphical abstract: -- Highlights: •Recent trends in the structural characterization of biodegradable polymers by MALDI and ESI MS are discussed. •MALDI MS as a noteworthy tool to follow the synthetic polymerization route of biodegradable materials is evidenced. •Elucidation of degradation mechanisms by modern MS techniques is examined. •ESI MS and HPLC–ESI MS are highlighted as highly suitable methods for structural and quantitative analysis of water-soluble biodegradation products. •Novel MS methods developed ad hoc and new MALDI matrices for biodegradable polymers are reviewed. -- Abstract: In the last decades, the solid-waste management related to the extensively growing production of plastic materials, in concert with their durability, have stimulated increasing interest in biodegradable polymers. At present, a variety of biodegradable polymers has already been introduced onto the market and can now be competitive with non biodegradable thermoplastics in different fields (packaging, biomedical, textile, etc.). However, a significant economical effort is still directed in tailoring structural properties in order to further broaden the range of applications without impairing biodegradation. Improving the performance of biodegradable materials requires a good characterization of both physico-chemical and mechanical parameters. Polymer analysis can involve many different features including detailed characterization of chemical structures and compositions as well as average molecular mass determination. It is of outstanding importance in troubleshooting of a polymer manufacturing process and for quality control, especially in biomedical applications. This review describes recent trends in the structural characterization of biodegradable materials by modern mass spectrometry (MS). It provides an overview of the analytical tools used to evaluate their degradation. Several successful applications of MALDI-TOF MS (matrix assisted laser desorption ionization

  13. Physiological and phylogenetic characterization of a stable chlorate-reducing benzene-degrading microbial community

    NARCIS (Netherlands)

    Weelink, S.A.B.; Tan, N.C.G.; Broeke, ten H.; Doesburg, van W.C.J.; Langenhoff, A.A.M.; Gerritse, J.; Stams, A.J.M.

    2007-01-01

    stable anoxic enrichment culture was obtained that degraded benzene with chlorate as an electron acceptor. The benzene degradation rate was 1.65 mM benzene per day, which is similar to reported aerobic benzene degradation rates but 20¿1650 times higher than reported for anaerobic benzene degradation

  14. Characterization of the juvenile hormone pathway in the viviparous cockroach, Diploptera punctata.

    Directory of Open Access Journals (Sweden)

    Juan Huang

    Full Text Available Juvenile hormones (JHs are key regulators of insect development and reproduction. The JH biosynthetic pathway is known to involve 13 discrete enzymatic steps. In the present study, we have characterized the JH biosynthetic pathway in the cockroach Diploptera punctata. The effect of exogenous JH precursors on JH biosynthesis was also determined. Based on sequence similarity, orthologs for the genes directly involved in the pathway were cloned, and their spatial and temporal transcript profiles were determined. The effect of shutting down the JH pathway in adult female cockroaches was studied by knocking down genes encoding HMG-CoA reductase (HMGR and Juvenile hormone acid methyltransferase (JHAMT. As a result, oocyte development slowed as a consequence of reduction in JH biosynthesis. Oocyte length, fat body transcription of Vg and ovarian vitellin content significantly decreased. In addition, silencing HMGR and JHAMT resulted in a decrease in the transcript levels of other genes in the pathway.

  15. Synthesis and thermal degradation characterization of novel poly(phosphazene-aryl amides

    Directory of Open Access Journals (Sweden)

    Z. P. Zhao

    2012-04-01

    Full Text Available New fully aromatic poly(phosphazene-aryl amides were prepared by polycondensation reaction of our synthesized aromatic diamine: 1,1,3,5-tetraphenoxy-4,6-bis(4-aminophenoxyoligocyclotriphosphazene (monomer 1 with terephthaloyl dichloride. Their chemical structure and composition were characterized by elemental analysis, 1H and 31P NMR (Nuclear Magnetic Resonance, and FT-IR (Fourier transform infrared spectroscopy, whereas their thermal degradation properties were determined by DSC (Differential Scanning Calorimetry and TGA (Thermal Gravimertic Analysis techniques. The solid residues of all samples were analysed by FT-IR and SEM (Scanning Electron Microscopy. Compared to conventional PPTA (poly(p-phenylene terephthamide, PPAA (poly(phosphazene-aryl amide shows excellent thermal stability and solubility in polar protic solvents. All poly(phosphazene-aryl amides show two thermal degradation in the temperature range 150–600°C. The monomer 1, due to its structure, shows the first maximum rate of thermal decomposition temperature around 150–350°C, which may be due to the decomposition of the P–O–C bone. Morphology of the solid residues by Scanning Electron Microscope exhibit that the granular of the solid residues gradual disappearance with the increase of monomer 1 content. The surface layer of PPAA solid residues has been grumous, for the syneresis of P–O–P took place.

  16. Preparation and characterization of tungsten-loaded titanium dioxide photocatalyst for enhanced dye degradation.

    Science.gov (United States)

    Saepurahman; Abdullah, M A; Chong, F K

    2010-04-15

    Tungsten-loaded TiO(2) photocatalyst has been successfully prepared and characterized. TEM analysis showed that the photocatalysts were nanosize with the tungsten species forming layers of coverage on the surface of TiO(2), but not in clustered form. This was confirmed by XRD and FT-Raman analyses where tungsten species were well dispersed at lower loading (12 mol%). In addition, loading with tungsten could stabilize the anatase phase from transforming into inactive rutile phase and did not shift the optical absorption to the visible region as shown by DRUV-vis analysis. PZC value of TiO(2) was found at 6.4, but the presence of tungsten at 6.5 mol% WO(3), decreased the PZC value to 3. Tungsten-loaded TiO(2) was superior to unmodified TiO(2) with 2-fold increase in degradation rate of methylene blue, and equally effective for the degradation of different class of dyes such as methyl violet and methyl orange at 1 mol% WO(3) loading.

  17. Synthesis and Characterization of Cerium Doped Titanium Catalyst for the Degradation of Nitrobenzene Using Visible Light

    Directory of Open Access Journals (Sweden)

    Padmini Ellappan

    2014-01-01

    Full Text Available Cerium doped catalyst was synthesized using Titanium isopropoxide as the Titanium source. The metal doped nanoparticles semiconductor catalyst was prepared by sol-sol method with the sol of Cerium. The synthesized catalyst samples were characterized by powder X-ray diffraction, BET surface area, thermogravimetric analysis (TGA, scanning electron microscopy (SEM, and UV-vis diffuse reflectance measurements (DRS and compared with undoped TiO2 catalyst. The photocatalytic activity of the sample was investigated for the decomposition of nitrobenzene (NB using visible light as the artificial light source. Cerium doped catalyst was found to have better degradation of nitrobenzene owing to its shift in the band gap from UV to visible region as compared to undoped TiO2 catalyst. The operational parameters were optimized with catalyst dosage of 0.1 g L−1, pH of 9, and light intensity of 500 W. The degradation mechanism followed the Langmuir Hinshelwood kinetic model with the rate constant depending nonlinearly on the operational parameters as given by the relationship Kapp (theoretical = 2.29 * 10−4 * Intensity0.584 * Concentration−0.230 * Dosage0.425 * pH0.336.

  18. Transition Metal Complexes of Naproxen: Synthesis, Characterization, Forced Degradation Studies, and Analytical Method Verification

    Directory of Open Access Journals (Sweden)

    Md. Sharif Hasan

    2016-01-01

    Full Text Available The aim of our current research was to synthesize some transition metal complexes of Naproxen, determine their physical properties, and examine their relative stability under various conditions. Characterizations of these complexes were done by 1H-NMR, Differential Scanning Calorimetry (DSC, FT-IR, HPLC, and scanning electron microscope (SEM. Complexes were subjected to acidic, basic, and aqueous hydrolysis as well as oxidation, reduction, and thermal degradation. Also the reversed phase high-performance liquid chromatography (RP-HPLC method of Naproxen outlined in USP was verified for the Naproxen-metal complexes, with respect to accuracy, precision, solution stability, robustness, and system suitability. The melting points of the complexes were higher than that of the parent drug molecule suggesting their thermal stability. In forced degradation study, complexes were found more stable than the Naproxen itself in all conditions: acidic, basic, oxidation, and reduction media. All the HPLC verification parameters were found within the acceptable value. Therefore, it can be concluded from the study that the metal complexes of Naproxen can be more stable drug entity and offer better efficacy and longer shelf life than the parent Naproxen.

  19. Synthesis and Characterization of Highly Efficient Nickel Nanocatalysts and Their Use in Degradation of Organic Dyes

    Directory of Open Access Journals (Sweden)

    Nazar Hussain Kalwar

    2014-01-01

    Full Text Available The present study describes the synthesis of highly active and ordered structures of nickel nanocatalysts by a facile, green, and economically viable approach. The study reveals efficient catalytic activity for the degradation of a number of toxic organic dyes, such as eosin-B (EB, rose bengal (RB, eriochrome black-T (ECBT, and methylene blue (MB. The stable ordered nickel nanostructure (Ni NSs arrays were prepared via a modified hydrazine reduction route with unique and controlled morphologies in a lyotropic liquid crystalline medium using a nonionic surfactant (Triton X-100. Characterization and optimization studies for the fabricated Ni NSs involving their surface binding interactions, size, and morphologies were carried out using UV-Vis spectroscopy, Fourier transform infrared (FTIR spectroscopy, X-ray diffraction (XRD, and scanning electron microscopy (SEM.

  20. Synthesis, spectral characterization and eukaryotic DNA degradation of thiosemicarbazones and their platinum(IV) complexes

    Science.gov (United States)

    Al-Hazmi, G. A.; El-Metwally, N. M.; El-Gammal, O. A.; El-Asmy, A. A.

    2008-01-01

    The condensation products of acetophenone (or its derivatives), salicylaldehyde and o-hydroxy- p-methoxybenzophenone with thiosemicarbazide and ethyl- or phenyl-thiosemicarbazide are the investigated thiosemicarbazones. Their reactions with H 2PtCl 6 produced Pt(IV) complexes characterized by elemental, thermal, mass, IR and electronic spectral studies. The coordination modes were found mononegative bidentate in the acetophenone derivatives and binegative tridentate in the salicylaldehyde derivatives. The complexes were analyzed thermogravimetrically and found highly stable. Some ligands and their complexes were screened against Sarcina sp. and E. coli using the cup-diffusion technique. [Pt( oHAT)(OH)Cl] shows higher activity against E. coli than the other compounds. The degradation power of the tested compounds on the calf thymus DNA supports their selectivity against bacteria and not against the human or related eukaryotic organisms.

  1. Hydrothermal synthesis of Ni(12)P(5) hollow microspheres, characterization and photocatalytic degradation property.

    Science.gov (United States)

    Li, Jun; Ni, Yonghong; Liao, Kaiming; Hong, Jianming

    2009-04-01

    In this paper, we report the successful synthesis of Ni(12)P(5) hollow spheres via a facile hydrothermal route, employing white phosphorus (WP) and nickel nitrate as the reactants in the presence of hexamethylenetetramine (HMT) and polyethylene glycol 10000 (PEG-10000). The phase and morphology of the product were characterized by means of powder X-ray diffraction (XRD), energy dispersive spectrometry (EDS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). HMT and surfactant (PEG-10000) played important roles in the formation of Ni(12)P(5) hollow microspheres. Furthermore, research also showed that the as-prepared Ni(12)P(5) hollow spheres could photocatalytically degrade some organic dyes such as Safranine T and Pyronine B under irradiation of 365 nm UV light.

  2. A putrescine-inducible pathway comprising PuuE-YneI in which gamma-aminobutyrate is degraded into succinate in Escherichia coli K-12.

    Science.gov (United States)

    Kurihara, Shin; Kato, Kenji; Asada, Kei; Kumagai, Hidehiko; Suzuki, Hideyuki

    2010-09-01

    Gamma-aminobutyrate (GABA) is metabolized to succinic semialdehyde by GABA aminotransferase (GABA-AT), and the succinic semialdehyde is subsequently oxidized to succinate by succinic semialdehyde dehydrogenase (SSADH). In Escherichia coli, there are duplicate GABA-ATs (GabT and PuuE) and duplicate SSADHs (GabD and YneI). While GabT and GabD have been well studied previously, the characterization and expression analysis of PuuE and YneI are yet to be investigated. By analyzing the amino acid profiles in cells of DeltapuuE and/or DeltagabT mutants, this study demonstrated that PuuE plays an important role in GABA metabolism in E. coli cells. The similarity of the amino acid sequences of PuuE and GabT is 67.4%, and it was biochemically demonstrated that the catalytic center of GabT is conserved as an amino acid residue important for the enzymatic activity in PuuE as Lys-247. However, the regulation of expression of PuuE is significantly different from that of GabT. PuuE is induced by the addition of putrescine to the medium and is repressed by succinate and low aeration conditions; in contrast, GabT is almost constitutive. Similarly, YneI is induced by putrescine, while GabD is not. For E. coli, PuuE is important for utilization of putrescine as a sole nitrogen source and both PuuE and YneI are important for utilization of putrescine as a sole carbon source. The results demonstrate that the PuuE-YneI pathway was a putrescine-inducible GABA degradation pathway for utilizing putrescine as a nutrient source.

  3. Identification of hydrologic and geochemical pathways using high frequency sampling, REE aqueous sampling and soil characterization at Koiliaris Critical Zone Observatory, Crete

    Energy Technology Data Exchange (ETDEWEB)

    Moraetis, Daniel, E-mail: moraetis@mred.tuc.gr [Department of Environmental Engineering, Technical University of Crete, 73100 Chania (Greece); Stamati, Fotini; Kotronakis, Manolis; Fragia, Tasoula; Paranychnianakis, Nikolaos; Nikolaidis, Nikolaos P. [Department of Environmental Engineering, Technical University of Crete, 73100 Chania (Greece)

    2011-06-15

    Highlights: > Identification of hydrological and geochemical pathways within a complex watershed. > Water increased N-NO{sub 3} concentration and E.C. values during flash flood events. > Soil degradation and impact on water infiltration within the Koiliaris watershed. > Analysis of Rare Earth Elements in water bodies for identification of karstic water. - Abstract: Koiliaris River watershed is a Critical Zone Observatory that represents severely degraded soils due to intensive agricultural activities and biophysical factors. It has typical Mediterranean soils under the imminent threat of desertification which is expected to intensify due to projected climate change. High frequency hydro-chemical monitoring with targeted sampling for Rare Earth Elements (REE) analysis of different water bodies and geochemical characterization of soils were used for the identification of hydrologic and geochemical pathways. The high frequency monitoring of water chemical data highlighted the chemical alterations of water in Koiliaris River during flash flood events. Soil physical and chemical characterization surveys were used to identify erodibility patterns within the watershed and the influence of soils on surface and ground water chemistry. The methodology presented can be used to identify the impacts of degraded soils to surface and ground water quality as well as in the design of methods to minimize the impacts of land use practices.

  4. Modern mass spectrometry in the characterization and degradation of biodegradable polymers.

    Science.gov (United States)

    Rizzarelli, Paola; Carroccio, Sabrina

    2014-01-15

    In the last decades, the solid-waste management related to the extensively growing production of plastic materials, in concert with their durability, have stimulated increasing interest in biodegradable polymers. At present, a variety of biodegradable polymers has already been introduced onto the market and can now be competitive with non biodegradable thermoplastics in different fields (packaging, biomedical, textile, etc.). However, a significant economical effort is still directed in tailoring structural properties in order to further broaden the range of applications without impairing biodegradation. Improving the performance of biodegradable materials requires a good characterization of both physico-chemical and mechanical parameters. Polymer analysis can involve many different features including detailed characterization of chemical structures and compositions as well as average molecular mass determination. It is of outstanding importance in troubleshooting of a polymer manufacturing process and for quality control, especially in biomedical applications. This review describes recent trends in the structural characterization of biodegradable materials by modern mass spectrometry (MS). It provides an overview of the analytical tools used to evaluate their degradation. Several successful applications of MALDI-TOF MS (matrix assisted laser desorption ionization time of flight) and ESI MS (electrospray mass spectrometry) for the determination of the structural architecture of biodegradable macromolecules, including their topology, composition, chemical structure of the end groups have been reported. However, MS methodologies have been recently applied to evaluate the biodegradation of polymeric materials. ESI MS represents the most useful technique for characterizing water-soluble polymers possessing different end group structures, with the advantage of being easily interfaced with solution-based separation techniques such as high-performance liquid

  5. Experimental Priapism is Associated with Increased Oxidative Stress and Activation of Protein Degradation Pathways in Corporal Tissue

    Science.gov (United States)

    Kanika, Nirmala D.; Melman, Arnold; Davies, Kelvin P.

    2010-01-01

    Priapism is a debilitating disease for which there is at present no clinically accepted pharmacologic intervention. It has been estimated that priapism lasting more than 24 hours in patients is associated with a 44–90% rate of erectile dysfunction (ED). In this investigation we determined in two animal models of priapism (opiorpin-induced priapism in the rat and priapism in a mouse model of sickle cell disease) if there is evidence for an increase in markers of oxidative stress in corporal tissue. In both animal models we demonstrate that priapism results in increased levels of lipid peroxidation, glutathione S-transferase activity, and oxidatively damaged proteins in corporal tissue. Using Western blot analysis we demonstrated there is up regulation of the ubiquitination ligase proteins, Nedd-4 and Mdm-2, and the lysososomal autophage protein, LC3. The anti-apoptotic protein, Bcl-2, was also up regulated. Overall, we demonstrate that priapism is associated with increased oxidative stress in corporal tissue and the activation of protein degradation pathways. Since oxidative stress is known to mediate the development of ED resulting from several etiologies (for example ED resulting from diabetes and aging) we suggest that damage to erectile tissue resulting from priapism might be prevented by treatments targeting oxidative stress. PMID:21085184

  6. Effects of rapid or slow body weight reduction on intramuscular protein degradation pathways during equivalent weight loss on rats.

    Science.gov (United States)

    Nonaka, Y; Urashima, S; Inai, M; Nishimura, S; Higashida, K; Terada, S

    2017-07-18

    The purpose of this study was to compare the effects of short-term fasting-induced rapid weight loss with those of slower but equivalent body weight loss induced by daily calorie restriction on muscle protein degradation pathways and muscle protein content. Male Fischer rats were subjected to either 30 % calorie restriction for 2 weeks to slowly decrease body weight (Slow) or 3-day fasting to rapidly decrease body weight by a comparable level of that of the Slow group (Rapid). The final body weights were about 15 % lower in both the Slow and Rapid groups than in the Con group (pweight of fast-twitch plantaris muscle, but not slow-twitch soleus muscle, were significantly lower in the Rapid group compared with the control rats fed ad libitum. Substantial increases in the expression ratio of autophagosomal membrane proteins (LC3-II/-I ratio) and polyubiquitinated protein concentration, used as biomarkers of autophagy-lysosome and ubiquitin-proteasome activities, respectively, were observed in the plantaris muscle of the Rapid group. Moreover, the LC3-II/-I ratio and polyubiquitinated protein concentration were negatively correlated with the total protein content and wet weight of plantaris muscle. These results suggest that short-term fasting-induced rapid body weight loss activates autophagy-lysosome and ubiquitin-proteasome systems more strongly than calorie restriction-induced slower weight reduction, resulting in muscular atrophy in fast-twitch muscle.

  7. RINL, guanine nucleotide exchange factor Rab5-subfamily, is involved in the EphA8-degradation pathway with odin.

    Directory of Open Access Journals (Sweden)

    Hiroaki Kajiho

    Full Text Available The Rab family of small guanosine triphosphatases (GTPases plays a vital role in membrane trafficking. Its active GTP-bound state is driven by guanine nucleotide-exchange factors (GEFs. Ras and Rab interactor (or Ras interaction/interference-like (RINL, which contains a conserved VPS9 domain critical for GEF action, was recently identified as a new Rab5 subfamily GEF in vitro. However, its detailed function and interacting molecules have not yet been fully elucidated. Here we found that RINL has GEF activity for the Rab5 subfamily proteins by measuring their GTP-bound forms in cultured cells. We also found that RINL interacts with odin, a member of the ankyrin-repeat and sterile-alpha motif (SAM domain-containing (Anks protein family. In addition, the Eph tyrosine kinase receptor EphA8 formed a ternary complex with both RINL and odin. Interestingly, RINL expression in cultured cells reduced EphA8 levels in a manner dependent on both its GEF activity and interaction with odin. In addition, knockdown of RINL increased EphA8 level in HeLa cells. Our findings suggest that RINL, as a GEF for Rab5 subfamily, is implicated in the EphA8-degradation pathway via its interaction with odin.

  8. Degradation of ethyl paraben by heat-activated persulfate oxidation: statistical evaluation of operating factors and transformation pathways.

    Science.gov (United States)

    Frontistis, Zacharias; Antonopoulou, Maria; Konstantinou, Ioannis; Mantzavinos, Dionissios

    2017-01-01

    A factorial design methodology was implemented to evaluate the importance of ethyl paraben (EP) concentration (500-1500 μg/L), sodium persulfate concentration (400-500 mg/L), temperature (40-60 °C), reaction time (2-30 min), water matrix (pure water or secondary treated wastewater), and initial solution pH (3-9) on EP removal by heat-activated persulfate oxidation. All individual effects, except the solution pH, were statistically significant and so were the second-order interactions of ethyl paraben concentration with temperature or the reaction time. The influence of the water matrix was crucial, and the efficiency of the process was lower in secondary treated wastewater due to the presence of natural organic matter and inorganic salts that compete with ethyl paraben for the reactive oxygen species. Liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) was employed to identify major transformation by-products (TBPs); 13 compounds were detected as TBPs of EP. Degradation occurred through (i) hydroxylation, (ii) dealkylation, and (iii) oligomerization reactions leading to TBPs with ether and biphenyl structures. Oligomerization reactions were found to be the dominant pathway during the first steps of the reaction. The toxicity of 500 μg/L EP in secondary treated wastewater was tested against marine bacteria Vibrio fischeri; toxicity increased during the first minutes due to the production of several TBPs, but it consistently decreased thereafter.

  9. Laser induced breakdown spectroscopy for analysis and characterization of degradation pathologies of Roman glasses

    Energy Technology Data Exchange (ETDEWEB)

    Palomar, T. [Instituto de Historia, Centro de Ciencias Humanas y Sociales, CSIC, C/Albasanz 26-28, 28037 Madrid (Spain); Oujja, M., E-mail: m.oujja@iqfr.csic.es [Instituto de Química Física Rocasolano, CSIC, C/Serrano 119, 28006 Madrid (Spain); García-Heras, M.; Villegas, M.A. [Instituto de Historia, Centro de Ciencias Humanas y Sociales, CSIC, C/Albasanz 26-28, 28037 Madrid (Spain); Castillejo, M. [Instituto de Química Física Rocasolano, CSIC, C/Serrano 119, 28006 Madrid (Spain)

    2013-09-01

    The feasibility and possibilities of laser induced breakdown spectroscopy (LIBS) in the full study of non-destructible historic glasses have been explored in the present work. Thirteen Roman glass samples, including seven entire glass beads, from the ancient town of Augusta Emerita (SW Spain) were characterized by LIBS in combination with other conventional techniques, such as scanning electron microscopy/energy dispersive X-ray spectrometry, X-ray fluorescence and ultraviolet–visible spectrophotometry. LIBS stratigraphic analysis, carried out by the application of successive laser pulses on the same spot, has been mainly targeted at characterizing particular features of non-destructible historic glasses, such as bulk chemical composition, surface degradation pathologies (dealkalinization layers and deposits), chromophores, and opacifying elements. The obtained data demonstrate that LIBS can be a useful and alternative technique for spectroscopic studies of historical glasses, especially for those conserved under burial conditions and when it deals with studying non-destructible samples. - Highlights: • Determination of chromophores and opacifiers in non-destructible glass by LIBS • Manganese is determined as principal component of dark deposits. • Antimony appears in all decorations while lead is only present in yellow ones. • Stratigraphic analysis enables the identification of dealkalinization layers.

  10. Characterizing Degradation Gradients through Land Cover Change Analysis in Rural Eastern Cape, South Africa

    Directory of Open Access Journals (Sweden)

    Zahn Münch

    2017-02-01

    Full Text Available Land cover change analysis was performed for three catchments in the rural Eastern Cape, South Africa, for two time steps (2000 and 2014, to characterize landscape conversion trajectories for sustained landscape health. Land cover maps were derived: (1 from existing data (2000; and (2 through object-based image analysis (2014 of Landsat 8 imagery. Land cover change analysis was facilitated using land cover labels developed to identify landscape change trajectories. Land cover labels assigned to each intersection of the land cover maps at the two time steps provide a thematic representation of the spatial distribution of change. While land use patterns are characterized by high persistence (77%, the expansion of urban areas and agriculture has occurred predominantly at the expense of grassland. The persistence and intensification of natural or invaded wooded areas were identified as a degradation gradient within the landscape, which amounted to almost 10% of the study area. The challenge remains to determine significant signals in the landscape that are not artefacts of error in the underlying input data or scale of analysis. Systematic change analysis and accurate uncertainty reporting can potentially address these issues to produce authentic output for further modelling.

  11. Biodegradability of HCH in agricultural soils from Guadeloupe (French West Indies): identification of the lin genes involved in the HCH degradation pathway.

    Science.gov (United States)

    Laquitaine, L; Durimel, A; de Alencastro, L F; Jean-Marius, C; Gros, O; Gaspard, S

    2016-01-01

    Banana has been a main agricultural product in the French West Indies (Guadeloupe and Martinique) since the 1960s. This crop requires the intensive use of pesticides to prevent attacks by insect pests. Chlorinated pesticides, such as hexachlorocyclohexane (HCH), chlordecone and dieldrin, were used until the beginning of the 1990s, resulting in a generalized diffuse contamination of the soil and water in the areas of banana production, hence the need to develop solutions for cleanup of the polluted sites. The aims of this work were (i) to assess lindane degradation in soil slurry microcosms treated with lindane at 10 mg/L and (ii) to detect the catabolic genes involved in the HCH degradation pathway. The soil slurry microcosm system showed a 40% lindane degradation efficiency at the end of a 30-day experiment. Lower lindane removal was also detected in the abiotic controls, probably caused by pesticide adsorption to soil particles. Indeed, the lindane concentration decreased from 6000 to 1330 ng/mL and from 800 to 340 ng/mL for the biotic and abiotic soils, respectively. Nevertheless, some of the genes involved in the HCH degradation pathway were amplified by polymerase chain reaction (PCR) from crude deoxyribonucleic acid (DNA) extracted from the Guadeloupe agricultural soil, suggesting that HCH degradation is probably mediated by bacteria closely related to the family Sphingomonadaceae.

  12. Development of a stability-indicating HPLC method of etifoxine with characterization of degradation products by LC-MS/TOF, 1H and 13C NMR.

    Science.gov (United States)

    Djabrouhou, Nadia; Guermouche, Moulay-Hassane

    2014-11-01

    This paper describes a new LC-MS/TOF method for the degradation products determination when Etifoxine (ETI) is submitted to different stress conditions. Chromatography is performed by using Kromasil C18 column (250mm×4.6mm, 5μm particle size). The selected mobile phase consists of formate buffer 0.02M, pH 3 and methanol (70/30, v/v). ETI is submitted to oxidative, acidic, basic, hydrolytic, thermal and UV light degradations. Detection is made at 254nm by photodiode array detector and mass spectrometry. A number of degradation products (DPs) called DPA, DPB, DPC and DPD are found depending on the stress; DPA with heat, DPA and DPB in acidic media or under UV-light; DPA, DPB and DPC under basic stress; DPA, DPB, DPC and DPD with oxidation. LC-MS/TOF is used to characterize the four DPs of ETI resulting from different stress conditions. (1)H and (13)C NMR are used to confirm the DP structures. The ETI fragmentation pathway is proposed. The method is validated with reference to International Conference on Harmonization guidelines and ETI are selectively determined in presence of its DPs, demonstrating its stability-indicating nature. Finally, for the validation step, specificity, linearity, accuracy and precision are determined for ETI and its DPs.

  13. A novel UV degradation product of Ebastine: isolation and characterization using Q-TOF, NMR, IR and computational chemistry.

    Science.gov (United States)

    Rapolu, Ravi; Pandey, Avadhesh Kumar; Raju, Ch Krishnam; Ghosh, Kaushik; Srinivas, Kolupula; Awasthi, Atul; Navalgund, Sameer G; Surendranath, Koduru V

    2015-03-25

    Forced degradation of Ebastine (1-(4-(1,1-dimethylethyl)phenyl)-4-(4-(diphenylmethoxy) piperidin-1-yl)butan-1-one) drug substance in ultraviolet light condition resulted into an unknown significant degradation product. This degradation product was analyzed using a newly developed reverse-phase HPLC, where it was eluted at 2.73 relative retention time to Ebastine peak. UV degradation product was isolated from reaction mass using preparative HPLC and its structure was elucidated using high resolution MS, multidimensional NMR and FTIR spectroscopic techniques. UV degradation product has been characterized as 2-(4-(benzhydryloxy)piperidin-1-yl)-1-(4-(tert-butyl)phenyl)-2-methylcyclopropanol. (1)H and (13)C NMR chemical shift values were generated using computational chemistry for possible two diastereomers (7R10S and 7R10R) and later 7R10R was confirmed (and its enantiomer) as final structure given it showed close agreement with experimental NMR data. Formation of UV degradation product as a recemic mixture was further verified by computational chemistry evaluation, chiral HPLC and polarimetery. To best of our knowledge, this is a novel degradation product which is not discussed at any form of publication yet.

  14. Aerobic degradation of polychlorinated biphenyls

    Energy Technology Data Exchange (ETDEWEB)

    Pieper, D.H. [Dept. of Environmental Microbiology, German Research Center for Biotechnology, Braunschweig (Germany)

    2005-04-01

    The microbial degradation of polychlorinated biphenyls (PCBs) has been extensively studied in recent years. The genetic organization of biphenyl catabolic genes has been elucidated in various groups of microorganisms, their structures have been analyzed with respect to their evolutionary relationships, and new information on mobile elements has become available. Key enzymes, specifically biphenyl 2,3-dioxygenases, have been intensively characterized, structure/sequence relationships have been determined and enzymes optimized for PCB transformation. However, due to the complex metabolic network responsible for PCB degradation, optimizing degradation by single bacterial species is necessarily limited. As PCBs are usually not mineralized by biphenyl-degrading organisms, and cometabolism can result in the formation of toxic metabolites, the degradation of chlorobenzoates has received special attention. A broad set of bacterial strategies to degrade chlorobenzoates has recently been elucidated, including new pathways for the degradation of chlorocatechols as central intermediates of various chloroaromatic catabolic pathways. To optimize PCB degradation in the environment beyond these metabolic limitations, enhancing degradation in the rhizosphere has been suggested, in addition to the application of surfactants to overcome bioavailability barriers. However, further research is necessary to understand the complex interactions between soil/sediment, pollutant, surfactant and microorganisms in different environments. (orig.)

  15. Small heat shock proteins target mutant cystic fibrosis transmembrane conductance regulator for degradation via a small ubiquitin-like modifier-dependent pathway.

    Science.gov (United States)

    Ahner, Annette; Gong, Xiaoyan; Schmidt, Bela Z; Peters, Kathryn W; Rabeh, Wael M; Thibodeau, Patrick H; Lukacs, Gergely L; Frizzell, Raymond A

    2013-01-01

    Small heat shock proteins (sHsps) bind destabilized proteins during cell stress and disease, but their physiological functions are less clear. We evaluated the impact of Hsp27, an sHsp expressed in airway epithelial cells, on the common protein misfolding mutant that is responsible for most cystic fibrosis. F508del cystic fibrosis transmembrane conductance regulator (CFTR), a well-studied protein that is subject to cytosolic quality control, selectively associated with Hsp27, whose overexpression preferentially targeted mutant CFTR to proteasomal degradation. Hsp27 interacted physically with Ubc9, the small ubiquitin-like modifier (SUMO) E2 conjugating enzyme, implying that F508del SUMOylation leads to its sHsp-mediated degradation. Enhancing or disabling the SUMO pathway increased or blocked Hsp27's ability to degrade mutant CFTR. Hsp27 promoted selective SUMOylation of F508del NBD1 in vitro and of full-length F508del CFTR in vivo, which preferred endogenous SUMO-2/3 paralogues that form poly-chains. The SUMO-targeted ubiquitin ligase (STUbL) RNF4 recognizes poly-SUMO chains to facilitate nuclear protein degradation. RNF4 overexpression elicited F508del degradation, whereas Hsp27 knockdown blocked RNF4's impact on mutant CFTR. Similarly, the ability of Hsp27 to degrade F508del CFTR was lost during overexpression of dominant-negative RNF4. These findings link sHsp-mediated F508del CFTR degradation to its SUMOylation and to STUbL-mediated targeting to the ubiquitin-proteasome system and thereby implicate this pathway in the disposal of an integral membrane protein.

  16. Activation of the cAMP/PKA pathway induces UT-A1 urea transporter monoubiquitination and targets it for lysosomal degradation.

    Science.gov (United States)

    Su, Hua; Chen, Minguang; Sands, Jeff M; Chen, Guangping

    2013-12-15

    Regulation of urea transporter UT-A1 in the kidney is important for the urinary concentrating mechanism. We previously reported that activation of the cAMP/PKA pathway by forskolin (FSK) leads to UT-A1 ubiquitination, endocytosis, and degradation. In this study, we discovered that FSK-induced UT-A1 ubiquitination is monoubiquitination as judged by immunoblotting with specific ubiquitin antibodies to the different linkages of the ubiquitin chain. UT-A1 monoubiquitination induced by FSK was processed mainly on the cell plasma membrane. Monoubiquitination facilitates UT-A1 endocytosis, and internalized UT-A1 is accumulated in the early endosome. Inhibition of ubiquitination by E1 ubiquitin-activating enzyme inhibitor PYR-41 significantly reduced FSK-induced UT-A1 endocytosis and degradation. Interestingly, FSK-stimulated UT-A1 degradation occurs through a lysosomal protein degradation system. We further found that the PKA phosphorylation sites of UT-A1 at Ser486 and Ser499 are required for FSK-induced UT-A1 monoubiquitination. The physiological significance was confirmed using rat kidney inner medullary collecting duct suspensions, which showed that vasopressin treatment promotes UT-A1 ubiquitination. We conclude that unlike under basal conditions in which UT-A1 is subject to polyubiquitination and proteasome-mediated protein degradation, activation of UT-A1 by FSK induces UT-A1 monoubiquitination and protein lysosomal degradation.

  17. Comparative metabolomics and structural characterizations illuminate colibactin pathway-dependent small molecules.

    Science.gov (United States)

    Vizcaino, Maria I; Engel, Philipp; Trautman, Eric; Crawford, Jason M

    2014-07-02

    The gene cluster responsible for synthesis of the unknown molecule "colibactin" has been identified in mutualistic and pathogenic Escherichia coli. The pathway endows its producer with a long-term persistence phenotype in the human bowel, a probiotic activity used in the treatment of ulcerative colitis, and a carcinogenic activity under host inflammatory conditions. To date, functional small molecules from this pathway have not been reported. Here we implemented a comparative metabolomics and targeted structural network analyses approach to identify a catalog of small molecules dependent on the colibactin pathway from the meningitis isolate E. coli IHE3034 and the probiotic E. coli Nissle 1917. The structures of 10 pathway-dependent small molecules are proposed based on structural characterizations and network relationships. The network will provide a roadmap for the structural and functional elucidation of a variety of other small molecules encoded by the pathway. From the characterized small molecule set, in vitro bacterial growth inhibitory and mammalian CNS receptor antagonist activities are presented.

  18. The ubiquitin+proteasome protein degradation pathway as a therapeutic strategy in the treatment of solid tumor malignancies.

    Science.gov (United States)

    Driscoll, James J; Minter, Alex; Driscoll, Daniel A; Burris, Jason K

    2011-02-01

    A concept that currently steers the development of cancer therapies has been that agents directed against specific proteins that facilitate tumorigenesis or maintain a malignant phenotype will have greater efficacy, less toxicity and a more sustained response relative to traditional cytotoxic chemotherapeutic agents. The clinical success of the targeted agent Imatinib mesylate as an inhibitor of the tyrosine kinase associated with the breakpoint cluster region-Abelson oncogene locus (BCR-ABL) in the treatment of Philadelphia-positive chronic myelogenous leukemia (CML) has served as a paradigm. While intellectually gratifying, the selective targeting of a single driver event by a small molecule, e.g., kinase inhibitor, to dampen a tumor-promoting pathway in the treatment of solid tumors is limited by many factors. Focus can alternatively be placed on targeting fundamental cellular processes that regulate multiple events, e.g., protein degradation, through the Ubiquitin (Ub)+Proteasome System (UPS). The UPS plays a critical role in modulating numerous cellular proteins to regulate cellular processes such as signal transduction, growth, proliferation, differentiation and apoptosis. Clinical success with the proteasome inhibitor bortezomib revolutionized treatment of B-cell lineage malignancies such as Multiple Myeloma (MM). However, many patients harbor primary resistance and do not respond to bortezomib and those that do respond inevitably develop resistance (secondary resistance). The lack of clinical efficacy of proteasome inhibitors in the treatment of solid tumors may be linked mechanistically to the resistance detected during treatment of hematologic malignancies. Potential mechanisms of resistance and means to improve the response to proteasome inhibitors in solid tumors are discussed.

  19. Photolysis of model emerging contaminants in ultra-pure water: kinetics, by-products formation and degradation pathways.

    Science.gov (United States)

    Benitez, F Javier; Acero, Juan L; Real, Francisco J; Roldan, Gloria; Rodriguez, Elena

    2013-02-01

    The photolysis of five frequent emerging contaminants (Benzotriazole, Chlorophene, N,N-diethyl-m-toluamide or DEET, Methylindole, and Nortriptyline HCl) was investigated in ultrapure water under monochromatic ultraviolet radiation at 254 nm and by a combination of UV and hydrogen peroxide. The results revealed that the photolysis rates followed first-order kinetics, with rate constant values depending on the nature of the specific compound, the pH, and the presence or absence of the scavenger tert-butanol. Quantum yields were also determined and values in the range of 53.8 × 10⁻³ - 9.4 × 10⁻³ mol E⁻¹ for Benzotriazole, 525 × 10⁻³ - 469 × 10⁻³ mol E⁻¹ for Chlorophene, 2.8 × 10⁻³ - 0.9 × 10⁻³ mol E⁻¹ for DEET, 108 × 10⁻³ - 165 × 10⁻³ mol E⁻¹ for Methylindole, and 13.8 × 10⁻³ - 15.0 × 10⁻³ mol E⁻¹ for Nortriptyline were obtained. The study also found that the UV/H₂O₂ process enhanced the oxidation rate in comparison to direct photolysis. High-performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS) technique was applied to the concentrations evaluation and further identification of the parent compounds and their by-products, which allowed the proposal of the degradation pathways for each compound. Finally, in order to assess the aquatic toxicity in the photodegradation of these compounds, the Vibrio fischeri acute toxicity test was used, and the results indicated an initial increase of this parameter in all cases, followed by a decrease in the specific case of Benzotriazole, DEET, Methylindole, and Chlorophene.

  20. GABA shunt and polyamine degradation pathway on γ-aminobutyric acid accumulation in germinating fava bean (Vicia faba L.) under hypoxia.

    Science.gov (United States)

    Yang, Runqiang; Guo, Qianghui; Gu, Zhenxin

    2013-01-01

    GABA shunt and polyamine degradation pathway on γ-aminobutyric acid (GABA) accumulation in germinating fava bean under hypoxia was investigated. GABA content, GAD and DAO activity were significantly increased under hypoxia treatment. Glu and polyamine contents enhanced largely and thus supplied as sufficient substrates for GABA formation. In contrast, GABA content decreased, mainly in the embryo, after removing the hypoxia stress. DAO activity, Glu and polyamines contents decreased, while an increment of GAD activity was observed. This indicated that GAD activity can be not only regulated by hypoxia, but by the rapid growth of embryo after the recovery from hypoxia stress. When treated with AG, DAO activity was almost inhibited completely, and the GABA content decreased by 32.96% and 32.07% after treated for 3 and 5 days, respectively. Hence, it can be inferred that about 30% of GABA formed in germinating fava bean under hypoxia was supplied by polyamine degradation pathway.

  1. Characterization of Clostridium thermocellum (B8) secretome and purified cellulosomes for lignocellulosic biomass degradation.

    Science.gov (United States)

    Osiro, Karen O; de Camargo, Brenda R; Satomi, Rachel; Hamann, Pedro Ricardo V; Silva, Jéssica Pinheiro; de Sousa, Marcelo Valle; Quirino, Betania F; Aquino, Elaine N; Felix, Carlos R; Murad, André Melro; Noronha, Eliane F

    2017-02-01

    The main goal of the present study was a complete proteomic characterization of total proteins eluted from residual substrate-bound proteins (RSBP), and cellulosomes secreted by Clostridium thermocellum B8 during growth in the presence of microcrystalline cellulose as a carbon source. The second goal was to evaluate their potential use as enzymatic blends for hydrolyzing agro-industrial residues to produce fermentable sugars. Protein identification through LC-MS/MS mass spectrometry showed that the RSBP sample, in addition to cellulosomal proteins, contains a wide variety of proteins, including those without a well-characterized role in plant cell wall degradation. The RSBP subsample defined as purified cellulosomes (PC) consists mainly of glycoside hydrolases grouped in families 5, 8, 9, 10 and 48. Dynamic light scattering, DLS, analysis of PC resulted in two protein peaks (pi1 and pi2) presenting molecular masses in agreement with those previously described for cellulosomes and polycellulosomes. These peaks weren't detected after PC treatment with 1.0% Tween. PC and RSBP presented maximal activities at temperatures ranging from 60° to 70°C and at pH 5.0. RSBP retained almost all of its activity after incubation at 50, 60 and 70°C and PC showed remarkable thermostability at 50 and 60°C. RSBP holocellullolytic activities were inhibited by phenolic compounds, while PC showed either increasing activity or a lesser degree of inhibition. RSBP and PC hydrolyze sugar cane straw, cotton waste and microcrystalline cellulose, liberating a diversity of saccharides; however, the highest concentration of released sugar was obtained for assays carried out using PC as an enzymatic blend and after ten days at 50°C. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Automated characterization of varnishes photo-degradation using portable T-controlled Raman spectroscopy

    Science.gov (United States)

    Osticioli, I.; Ciofini, D.; Mencaglia, A. A.; Siano, S.

    2017-02-01

    In this work, a portable-Raman device (excitation wavelength 1064 nm) was employed for the first time for continuously monitoring the complex molecular dynamics of terpenoid resins (dammar, mastic, colophony, sandarac and shellac), which occur during their ageing under artificial light exposure. The instrumentation was equipped with a pyroelectric sensor allowing for temperature control of the sample's irradiated surface while the acquisition of spectra occurs by setting fixed maximum temperature and total radiant exposure. Resins were dropped into special pits over a dedicated rotating wheel moved by a USB motor. The rotation allowed samples sliding between the positions designated for the acquisition of the Raman spectra and that for artificial ageing. Samples were exposed to artificial light for 45-days and almost 400 spectra for each resin sample were collected. The exposure to artificial light led to significant changes allowing the characterization of the alteration process. The automated acquisition of a large number of spectra overtime during light-exposure has given the possibility to distinguish fast dynamics, mainly associated to solvent evaporation, from those slower due to resins photo-degradation processes.

  3. Conjugates of degraded and oxidized hydroxyethyl starch and sulfonylureas: synthesis, characterization, and in vivo antidiabetic activity.

    Science.gov (United States)

    Abbas, Muhammad Azhar; Hameed, Shahid; Farman, Muhammad; Kressler, Jörg; Mahmood, Nasir

    2015-01-21

    Orally administered drugs usually face the problem of low water solubility, low permeability, and less retention in bloodstream leading to unsatisfactory pharmacokinetic profile of drugs. Polymer conjugation has attracted increasing interest in the pharmaceutical industry for delivering such low molecular weight (Mw) drugs as well as some complex compounds. In the present work, degraded and oxidized hydroxyethyl starch (HES), a highly biocompatible semisynthetic biopolymer, was used as a drug carrier to overcome the solubility and permeability problems. The HES was coupled with synthesized N-arylsulfonylbenzimidazolones, a class of sulfonylurea derivatives, by creating an amide linkage between the two species. The coupled products were characterized using GPC, FT-IR, (1)H NMR, and (13)C NMR spectroscopy. The experiments established the viability of covalent coupling between the biopolymer and N-arylsulfonylbenzimidazolones. The coupled products were screened for their in vivo antidiabetic potential on male albino rats. The coupling of sulfonylurea derivatives with HES resulted in a marked increase of the hypoglycemic activity of all the compounds. 2,3-Dihydro-3-(4-nitrobenzensulfonyl)-2-oxo-1H-benzimidazole coupled to HES10100 was found most potent with a 67% reduction in blood glucose level of the rats as compared to 41% reduction produced by tolbutamide and 38% by metformin.

  4. Isolation and characterization of a diverse group of phenylacetic acid degrading microorganisms from pristine soil.

    Science.gov (United States)

    O'Connor, Kevin E; O'Leary, Niall P; Marchesi, Julian R; Dobson, Alan D W; Duetz, Wouter

    2005-11-01

    A diverse range of microorganisms capable of growth on phenylacetic acid as the sole source of carbon and energy were isolated from soil. Sixty six different isolates were identified and grouped according to 16S rRNA gene RFLP analysis. Subsequent sequencing of 16S rDNA from selected strains allowed further characterization of the phenylacetic acid degrading population isolated from soil. Nearly half (30) of the isolates are Bacillus species while the rest of the isolates are strains from a variety of genera namely, Arthrobacter, Pseudomonas, Rhodococcus, Acinetobacter, Enterobacter, Flavobacterium, and Paenibacillus. Sixty-one of the sixty-six strains reproducibly grew in defined minimal liquid culture medium (E2). All strains isolated grew when at least one hydroxylated derivative of phenylacetic acid was supplied as the carbon source, while 59 out of the 61 strains tested, accumulated ortho-hydroxyphenylacetic acid in the assay buffer; when pulsed with phenylacetic acid. Oxygen consumption experiments failed to indicate a clear link between phenylacetic acid and hydroxy-substituted phenylacetic acid in isolates from a broad range of genera.

  5. Bismuth Modified Porous Silica Preparation, Characterization and Photocatalytic Activity Evaluation for Degradation of Isoproturon

    Institute of Scientific and Technical Information of China (English)

    Anil Kumar Reddy Police; Srinivas Basavaraju; Durga Kumari Valluri; Subrahmanyam Machiraju

    2013-01-01

    Porous silica prepared by using an acrylic emulsion has been impregnated with bismuth ion resulting in Bi2Sio5 species containing surface.The as-prepared materials have been characterized by X-ray diffraction spectroscopy (XRD),X-ray photoelectron spectroscopy (XPS),UV-Vis diffuse reflectance spectroscopy (UV-Vis DRS),scanning electron microscopy (SEM),energy dispersive analysis of X-ray (EDAX),transmission electron microscopy (TEM),Fourier transform infrared spectroscopy (FTIR) and N2 adsorption/desorption techniques.EDAX analysis confirms the penetration of bismuth ions into the framework of silica to form Bi2Sio5,which is substantiated by XRD.The UV-Vis DRS shows that the catalysts are optically active and XPS confirms the inclusion of bismuth into the framework of silica.FTIR spectra illustrate the formation of Bi-O-Si linkages in the porous silica framework.SEM and TEM show the spherical morphology,whereas N2 adsorption/desorption study confirms the porosity of the prepared materials.The photocatalytic activity of the material is evaluated for the degradation of isoproturon herbicide and it is found that the material is active as compared to the commercial P-25 Degussa Tio2.

  6. Synthesis and characterization of CNT/Ce-TiO2 nanocomposite for phenol degradation

    Institute of Scientific and Technical Information of China (English)

    Shaari N; Tan S H; Mohamed A R

    2012-01-01

    An innovative photocatalyst,Carbon nanotube (CNT) supported Ce-TiO2 nanocomposite was successfully synthesized via modified sol gel method and investigated in a batch reactor for abolition of phenol under UV light spectrum.Characterization of catalyst microstructure and internal properties were done by means of X-ray diffraction (XRD),Brunauer-Emmett-Teller (BET),scanning electron microscopy (SEM),transmission electron microscopy (TEM) and UV-vis diffuse reflectance spectra (DRS).Ce doping can inhibit phase transformation from anatase to rutile and eliminate the recombination of electron-hole pairs in the catalyst.The presence of CNT in TiO2 composite can both increase the photoactivity under UV and change surface properties to achieve sensitivity to visible light.The optimum mass ratio of CNT support and cerium (Ce) dopant in TiO2 was the prominent factor to harvest CNT/Ce-TiO2 photocatalyst nanocomposite.The results demonstrated that optimum mass ratio of CNT:TiO2:Ce was 0.02:1.0:0.06,which resulted in the great performance of the photocatalyst to degrade about 94% of phenol in a 50 mg/L solution in only 3 h.In this paper,dissimilar role of CNT support and Ce dopant in the TiO2 photocatalysis of phenol was also discussed.

  7. Characterization of enzymatically induced degradation of articular cartilage using high frequency ultrasound

    Science.gov (United States)

    Töyräs, J.; Rieppo, J.; Nieminen, M. T.; Helminen, H. J.; Jurvelin, J. S.

    1999-11-01

    Ultrasound may provide a quantitative technique for the characterization of cartilage changes typical of early osteoarthrosis. In this study, specific changes in bovine articular cartilage were induced using collagenase and chondroitinase ABC, enzymes that selectively degrade collagen fibril network and digest proteoglycans, respectively. Changes in cartilage structure and properties were quantified using high frequency ultrasound, microscopic analyses and mechanical indentation tests. The ultrasound reflection coefficient of the physiological saline-cartilage interface (R1) decreased significantly (-96.4%, p<0.01) in the collagenase digested cartilage compared to controls. Also a significantly lower ultrasound velocity (-6.2%, p<0.01) was revealed after collagenase digestion. After chondroitinase ABC digestion, a new acoustic interface at the depth of the enzyme penetration front was detected. Cartilage thickness, as determined with ultrasound, showed a high, linear correlation (R = 0.943, n = 60, average difference 0.073 mm (4.0%)) with the thickness measured by the needle-probe method. Both enzymes induced a significant decrease in the Young's modulus of cartilage (p<0.01). Our results indicate that high frequency ultrasound provides a sensitive technique for the analysis of cartilage structure and properties. Possibly ultrasound may be utilized in vivo as a quantitative probe during arthroscopy.

  8. Characterization of a trehalose-degrading enzyme from the hyperthermophilic archaeon Sulfolobus acidocaldarius.

    Science.gov (United States)

    Moon, Jeong Hyun; Lee, Whiso; Park, Jihee; Choi, Kyoung-Hwa; Cha, Jaeho

    2016-07-01

    We purified a cytosolic trehalase (TreH) from a thermoacidophilic archaeon Sulfolobus acidocaldarius. Enzyme activity in cell-free extracts indicated that trehalose degradation in the cell occurred via the hydrolytic activity of TreH, and not via TreP (phosphorolytic activity) or TreT (transfer activity). TreH was purified to near-homogeneity by DEAE anion-exchange chromatography, followed by size exclusion and HiTrap Q anion-exchange chromatography, and its molecular mass was estimated as 40 kDa. Maximum activity was observed at 85°C and pH 4.5. The half-life of TreH was 53 and 41 min at 90°C and 95°C, respectively. TreH was highly specific for trehalose and was inhibited by glucose with a Ki of 0.05 mM. Compared with TreH from other trehalases, TreH from S. acidocaldarius is the most thermostable trehalase reported so far. Furthermore, this is the first trehalase characterized in the Archaea domain.

  9. Isolation and characterization of agar-degrading endophytic bacteria from plants.

    Science.gov (United States)

    Song, Tao; Zhang, Weijia; Wei, Congchong; Jiang, Tengfei; Xu, Hui; Cao, Yi; Cao, Yu; Qiao, Dairong

    2015-02-01

    Agar is a polysaccharide extracted from the cell walls of some macro-algaes. Among the reported agarases, most of them come from marine environment. In order to better understand different sources of agarases, it is important to search new non-marine native ones. In this study, seven agar-degrading bacteria were first isolated from the tissues of plants, belonging to three genera, i.e., Paenibacillus sp., Pseudomonas sp., and Klebsiella sp. Among them, the genus Klebsiella was first reported to have agarolytic ability and the genus Pseudomonas was first isolated from non-marine environment with agarase activity. Besides, seven strains were characterized by investigating the growth and agarase production in the presence of various polysaccharides. The results showed that they could grow on several polysaccharides such as araban, carrageenan, chitin, starch, and xylan. Besides, they could also produce agarase in the presence of different polysaccharides other than agar. Extracellular agarases from seven strains were further analyzed by SDS-PAGE combined with activity staining and estimated to be 75 kDa which has great difference from most reported agarases.

  10. Boron-doped diamond electrode: Preparation, characterization and application for electrocatalytic degradation of m-dinitrobenzene.

    Science.gov (United States)

    Bai, Hongmei; He, Ping; Pan, Jing; Chen, Jingchao; Chen, Yang; Dong, Faqing; Li, Hong

    2017-07-01

    Boron-doped diamond (BDD) electrode was successfully prepared via microwave plasma chemical vapor deposition method and it was used in electrocatalytic degradation of m-dinitrobenzene (m-DNB). The electrocatalytic degradation efficiency of m-DNB was evaluated under different experimental parameters including current density, temperature, pH, Na2SO4 concentration and initial m-DNB concentration. Under optimal parameters, degradation efficiency of m-DNB reached up to 82.7% after 150min. The degradation process of m-DNB was fitted well with pseudo first-order kinetics. Moreover, UV and HPLC analyses implied that m-DNB was totally destroyed and mineralized after 240min degradation, and the proposed mechanism during the electrocatalytic degradation process was analyzed. All these results demonstrated that BDD electrode possessed excellent electrocatalytic property and showed a great potential application in wastewater treatment.

  11. Study of acaricide stability in honey. Characterization of amitraz degradation products in honey and beeswax.

    Science.gov (United States)

    Korta, E; Bakkali, A; Berrueta, L A; Gallo, B; Vicente, F; Kilchenmann, V; Bogdanov, S

    2001-12-01

    A study on the possible degradation of amitraz, bromopropylate, coumaphos, chlordimeform, cymiazole, flumethrin, and tau-fluvalinate during the storage of honey was carried out by HPLC. Except amitraz, the other acaricides are stable in this medium for at least 9 months. Degradation studies of amitraz in honey and beeswax were carried out; the degradation products detected in both matrices were 2,4-dimethylphenylformamide (DMF) and N-(2,4-dimethylphenyl)-N'-methylformamidine (DPMF). The reaction rate constants and the half-lives of the amitraz degradation in honey and wax were calculated. Amitraz was nearly completely degraded within 1 day in beeswax and within 10 days in honey. When amitraz-spiked combs are recycled into new beeswax, DMF was found to be the principal degradation product left in pure wax.

  12. Characterization of changes in gene expression and biochemical pathways at low levels of benzene exposure.

    Directory of Open Access Journals (Sweden)

    Reuben Thomas

    Full Text Available Benzene, a ubiquitous environmental pollutant, causes acute myeloid leukemia (AML. Recently, through transcriptome profiling of peripheral blood mononuclear cells (PBMC, we reported dose-dependent effects of benzene exposure on gene expression and biochemical pathways in 83 workers exposed across four airborne concentration ranges (from 10 ppm compared with 42 subjects with non-workplace ambient exposure levels. Here, we further characterize these dose-dependent effects with continuous benzene exposure in all 125 study subjects. We estimated air benzene exposure levels in the 42 environmentally-exposed subjects from their unmetabolized urinary benzene levels. We used a novel non-parametric, data-adaptive model selection method to estimate the change with dose in the expression of each gene. We describe non-parametric approaches to model pathway responses and used these to estimate the dose responses of the AML pathway and 4 other pathways of interest. The response patterns of majority of genes as captured by mean estimates of the first and second principal components of the dose-response for the five pathways and the profiles of 6 AML pathway response-representative genes (identified by clustering exhibited similar apparent supra-linear responses. Responses at or below 0.1 ppm benzene were observed for altered expression of AML pathway genes and CYP2E1. Together, these data show that benzene alters disease-relevant pathways and genes in a dose-dependent manner, with effects apparent at doses as low as 100 ppb in air. Studies with extensive exposure assessment of subjects exposed in the low-dose range between 10 ppb and 1 ppm are needed to confirm these findings.

  13. Structural Characterization of the Degradation Products of a Minor Natural Sweet Diterpene Glycoside Rebaudioside M under Acidic Conditions

    OpenAIRE

    Prakash, Indra; Chaturvedula, Venkata; Markosyan, Avetik

    2014-01-01

    Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0) and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C-) &

  14. Structural characterization of the degradation products of a minor natural sweet diterpene glycoside Rebaudioside M under acidic conditions.

    Science.gov (United States)

    Prakash, Indra; Chaturvedula, Venkata Sai Prakash; Markosyan, Avetik

    2014-01-14

    Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0) and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C-) & 2D (COSY, HSQC, HMBC) NMR, HRMS, MS/MS spectral data as well as enzymatic and acid hydrolysis studies.

  15. Structural Characterization of the Degradation Products of a Minor Natural Sweet Diterpene Glycoside Rebaudioside M under Acidic Conditions

    OpenAIRE

    Indra Prakash; Venkata Sai Prakash Chaturvedula; Avetik Markosyan

    2014-01-01

    Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0) and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C-) &...

  16. Structural Characterization of the Degradation Products of a Minor Natural Sweet Diterpene Glycoside Rebaudioside M under Acidic Conditions

    OpenAIRE

    Prakash, Indra; Chaturvedula, Venkata Sai Prakash; Markosyan, Avetik

    2014-01-01

    Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0) and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C-) & 2D ...

  17. Structural Characterization of the Degradation Products of a Minor Natural Sweet Diterpene Glycoside Rebaudioside M under Acidic Conditions

    Directory of Open Access Journals (Sweden)

    Indra Prakash

    2014-01-01

    Full Text Available Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0 and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C- & 2D (COSY, HSQC, HMBC NMR, HRMS, MS/MS spectral data as well as enzymatic and acid hydrolysis studies.

  18. Structural Characterization of the Degradation Products of a Minor Natural Sweet Diterpene Glycoside Rebaudioside M under Acidic Conditions

    Science.gov (United States)

    Prakash, Indra; Chaturvedula, Venkata Sai Prakash; Markosyan, Avetik

    2014-01-01

    Degradation of rebaudioside M, a minor sweet component of Stevia rebaudiana Bertoni, under conditions that simulated extreme pH and temperature conditions has been studied. Thus, rebaudioside M was treated with 0.1 M phosphoric acid solution (pH 2.0) and 80 °C temperature for 24 h. Experimental results indicated that rebaudioside M under low pH and higher temperature yielded three minor degradation compounds, whose structural characterization was performed on the basis of 1D (1H-, 13C-) & 2D (COSY, HSQC, HMBC) NMR, HRMS, MS/MS spectral data as well as enzymatic and acid hydrolysis studies. PMID:24424316

  19. [Research on the degradation of BaP with potassium ferrate characterized by fluorescence].

    Science.gov (United States)

    Chen, Yu-Zhe; He, Qiang; Yu, Dan-Ni; Li, Si; Tan, Xue-Mei

    2012-07-01

    The degradation of Benzo(a)pyrene (BaP) by potassium ferrate was researched by means of multiple fluorescence spectroscopic methods such as synchronous, time-scan, excitation emission matrix (EEM) and photometry, under the optimal condition. Within the degradation process, the characteristics of the BaP's concentration at different time-intervals, and the kinetics of the degradation of BaP by potassium ferrate were discussed. From the experimental data, both synchronous and EEM spectra's results showed that the concentration of BaP was reduced 90% by potassium ferrate within 20 s after degradation, and the reaction process was very slow after 60 s. The degradation kinetic equation, ln(F0/Ft) = 0.563 2t + 0.171 2, (R2 = 0.994 2), was obtained through a convenient and fast way combining the time-scan fluorescence data and photometry data, and the photometry included the synchronous photometry and emission photometry. According to the kinetic equation, the degradation of BaP by potassium ferrate was in accord with the order of the first order reaction. So this article could provide a very useful conference for the research on the pollutant degradation by potassium ferrate, especially for the degradation process and the degradation mechanisms.

  20. A stability indicating HPLC method for the determination of clobazam and its basic degradation product characterization

    OpenAIRE

    2014-01-01

    Background Clobazam is used for the treatment of different types of seizure and epilepsy. The present research is undertaken to study the systematic forced degradation of clobazam and to identify its main degradation product under basic conditions. Methods The degradation of clobazam was studied under different conditions. Clobazam and its degradation products were separated using a Nova-Pak C18 column and a mixture of KH2PO4 50 mM (pH 8.5) and acetonitrile (50:50, v/v) as the mobile phase wi...

  1. Analysis of neem oils by LC-MS and degradation kinetics of azadirachtin-A in a controlled environment. Characterization of degradation products by HPLC-MS-MS.

    Science.gov (United States)

    Barrek, Sami; Paisse, Olivier; Grenier-Loustalot, Marie-Florence

    2004-02-01

    Since it was first isolated, the oil extracted from seeds of neem (Azadirachtin indica A juss) has been extensively studied in terms of its efficacy as an insecticide. Several industrial formulations are produced as emulsifiable solutions containing a stated titer of the active ingredient azadirachtin-A (AZ-A). The work reported here is the characterization of a formulation of this insecticide marketed under the name of Neem-azal T/S and kinetic studies of the major active ingredient of this formulation. We initially performed liquid-liquid extraction to isolate the neem oil from other ingredients in the commercial mixture. This was followed by a purification using flash chromatography and semi-preparative chromatography, leading to (13)C NMR identification of structures such as azadirachtin-A, azadirachtin-B, and azadirachtin-H. The neem extract was also characterized by HPLC-MS using two ionization sources, APCI (atmospheric pressure chemical ionization) and ESI (electrospray ionization) in positive and negative ion modes of detection. This led to the identification of other compounds present in the extract-azadirachtin-D, azadirachtin-I, deacetylnimbin, deacetylsalannin, nimbin, and salannin. The comparative study of data gathered by use of the two ionization sources is discussed and shows that the ESI source enables the largest number of structures to be identified. In a second part, kinetic changes in the main product (AZ-A) were studied under precise conditions of pH (2, 4, 6, and 8), temperature (40 to 70 degrees C), and light (UV, dark room and in daylight). This enabled us to determine the degradation kinetics of the product (AZ-A) over time. The activation energy of the molecule (75+/-9 kJ mol(-1)) was determined by examining thermal stability in the range 40 to 70 degrees C. The degradation products of this compound were identified by use of HPLC-MS and HPLC-MS-MS. The results enabled proposal of a chemical degradation reaction route for AZ-A under

  2. Synthesis, Characterization, and Evaluation of Boron-Doped Iron Oxides for the Photocatalytic Degradation of Atrazine under Visible Light

    Directory of Open Access Journals (Sweden)

    Shan Hu

    2012-01-01

    Full Text Available Photocatalytic degradation of atrazine by boron-doped iron oxides under visible light irradiation was investigated. In this work, boron-doped goethite and hematite were successfully prepared by sol-gel method with trimethylborate as boron precursor. The powders were characterized by XRD, UV-vis diffuse reflectance spectra, and porosimetry analysis. The results showed that boron doping could influence the crystal structure, enlarge the BET surface area, improve light absorption ability, and narrow their band-gap energy. The photocatalytic activity of B-doped iron oxides was evaluated in the degradation of atrazine under the visible light irradiation, and B-doped iron oxides showed higher atrazine degradation rate than that of pristine iron oxides. Particularly, B-doped goethite exhibited better photocatalytic activity than B-doped hematite.

  3. LC-MS characterization of valsartan degradation products and comparison with LC-PDA

    Directory of Open Access Journals (Sweden)

    Sumaia Araújo Pires

    2015-12-01

    Full Text Available abstract Valsartan was submitted to forced degradation under acid hydrolysis condition as prescribed by the ICH. Degraded sample aliquots were separated via HPLC using a Hypersil ODS (C18 column (250 x 4.6 mm i.d., 5 µm. Either photodiode array (PDA detection or mass spectrometry (MS full scan monitoring of HPLC runs were used. HPLC-PDA failed to indicate Valsartan degradation under forced acid degradation, showing an insignificant peak area variation and that Valsartan apparently remained pure. HPLC-MS using electrospray ionization (ESI and total ionic current (TIC monitoring did not reveal any peak variation either, but inspection of the ESI mass spectra showed the appearance of m/z 306 and m/z 352 ions for the same retention time as that of Valsartan (m/z 436. These ions were identified as being protonated molecules of two co-eluting degradation products formed by hydrolysis. These assignments were confirmed by ESI-MS/MS with direct infusion of the degraded samples. The results showed that the use of selective HPLC-MS is essential for monitoring Valsartan degradation. Efficient HPLC separation coupled to selective and structural diagnostic MS monitoring seems therefore mandatory for comprehensive drug degradation studies, particularly for new drugs and formulations, and for method development.

  4. Transport and degradation of dissolved organic matter and associated freshwater pathways in the Laptev Sea (Siberian Arctic)

    Science.gov (United States)

    Hoelemann, Jens; Janout, Markus; Koch, Boris; Bauch, Dorothea; Hellmann, Sebastian; Eulenburg, Antje; Heim, Birgit; Kassens, Heidemarie; Timokhov, leonid

    2016-04-01

    The Siberian shelves are seasonally ice-covered and characterized by large freshwater runoff rates from some of the largest rivers on earth. These rivers also provide a considerable amount of dissolved organic carbon (DOC) to the Arctic Ocean. With an annual load of about 6 Tg DOC a-1 the Lena River contributes nearly 20 percent of the annual DOC discharge to the Arctic Ocean. We present a comprehensive dataset collected during multiple Laptev Sea expeditions carried out in spring, summer and fall (2010-15) in order to explore the processes controlling the dispersal and degradation of DOM during the river water's passage across the shelf. Our investigations are focused on CDOM (Colored Dissolved Organic Matter), which resembles the DOC concentration, interacts with solar radiation and forms a major fraction of the organic matter pool. Our results show an inverse correlation between salinity and CDOM, which emphasizes its terrigenous source. Further, the spectral slope of CDOM absorption indicates that photochemical bleaching is the main process that reduces the CDOM absorption (~ 20%) in freshwater along its transport across the shelf. The distribution of the Lena river water is primarily controlled by winds in summer. During summers with easterly or southerly winds, the plume remains on the central and northern Laptev shelf, and is available for export into the Arctic Basin. The CDOM-rich river water increases the absorption of solar radiation and enhances warming of a shallow surface layer. This emphasizes the importance of CDOM for sea surface temperatures and lateral ice melt on the shelf and adjacent basin. DOC concentrations in freshwater vary seasonally and become larger with increasing discharge. Our data indicate that the CDOM concentrations are highest during the freshet when landfast ice is still present. Subsequent mixing with local sea ice meltwater lowers CDOM to values that are characteristic for the Lena freshwater during the rest of the year.

  5. Cloning and expression of meta-cleavage enzyme (CarB of carbazole degradation pathway from Pseudomonas stutzeri

    Directory of Open Access Journals (Sweden)

    Ariane Leites Larentis

    2005-06-01

    Full Text Available In this work, the 1082bp PCR product corresponding to carBaBb genes that encode the heterotetrameric enzyme 2'-aminobiphenyl-2,3-diol 1,2-dioxygenase (CarB, involved in the Pseudomonas stutzeri ATCC 31258 carbazole degradation pathway, was cloned using the site-specific recombination system. Recombinant clones were confirmed by PCR, restriction enzyme digestion and sequencing. CarB dioxygenase was expressed in high levels and in active form in Escherichia coli BL21-SI using the His-tagged expression vector pDEST TM17 and salt induction for 4h.Carbazol e seus derivados são compostos nitrogenados aromáticos, presentes comumente em petróleo e potencialmente poluentes. A rota de biodegradação de carbazol a ácido antranílico em Pseudomonas sp. é composta por três enzimas responsáveis, respectivamente, pelas reações de dioxigenação angular, meta-clivagem e hidrólise. A segunda enzima da rota, 2'-aminobifenil-2,3-diol 1,2-dioxigenase (CarB, codificada por dois genes (carBa e carBb, é um heterotetrâmero com atividade catalítica na quebra do anel catecol do susbtrato na posição meta. Neste trabalho, foi clonado o produto de PCR de 1082pb correspondente aos genes carBaBb da bactéria degradadora de carbazol Pseudomonas stutzeri ATCC 31258. A estratégia de clonagem empregada foi a de recombinação sítio-específica e a construção dos plasmídeos foi confirmada por PCR, digestão com enzima de restrição e seqüenciamento. A enzima ativa foi expressa em altas concentrações em vetor pDEST TM17 com cauda de histidina e promotor T7 em Escherichia coli BL21-SI com indução por NaCl durante 4h.

  6. Isolation and characterization of Wnt pathway-related genes from Porifera.

    Science.gov (United States)

    Adell, Teresa; Thakur, Archana N; Müller, Werner E G

    2007-09-01

    The Wnt signal acts by binding to Frizzled receptors, with the subsequent activation of two different signal transduction cascades, the canonical and the non-canonical Wnt pathways, involved in cell growth, differentiation, migration and fate. The canonical pathway functions through the translocation of beta-catenin to the nucleus and the activation of TCF/LEF transcription factors; it plays an important role in developmental patterning and cell fate decisions during embryogenesis. The non-canonical Wnt pathway is responsible for the planar cell polarity process in invertebrates, and for the convergent-extension movements during vertebrate gastrulation. The final effect of the non-canonical Wnt pathway is the rearrangement of the cell cytoskeleton, through the activation of the subfamily of Ras-like small GTPases. In a recent report we described for the first time the isolation of a Wnt-related gene, Sd-Frizzled, from the most basal animal phylum, the Porifera. In the present study we report the isolation and phylogenetic characterization of several Wnt pathway-related genes from the sponge Suberites domuncula: Sd-TCF/LEF, Sd-GSK3, a recently discovered molecule with a putative function as a Wnt regulator (Sd-LZIC), the small Rho GTPases Sd-RhoA, Sd-Cdc42, and their effector Sd-mrlc. Also the isolation of a secreted frizzled related protein sFRP from another sponge species (Lubomirskia baicalensis) is reported.

  7. Isolation and Characterization of a Fucoidan-Degrading Bacterium from Laminaria japonica

    Institute of Scientific and Technical Information of China (English)

    WANG Ying; LI Bafang; ZHAO Xue; PIAO Meizi

    2014-01-01

    Fucoidan, a polysaccharide containing abundant fucose and sulfate ester group, was prepared from Laminaria japonica. In order to obtain fucoidan-degrading enzyme, bacteria capable of degrading fucoidan were screened from kelp. A bacterial strain named RC2-3 was obtained, which degraded fucoidan by the maximum extent of 54%± 1.3%, the highest among all bacterial isolates. High-performance size exclusion chromatography (HPSEC) showed that the molecular weight of fucoidan was gradually reduced by RC2-3 with culturing time, suggesting the production of fucoidan-degrading enzyme by RC2-3. Phylogenetic analysis of partial 16S ribosomal RNA gene (16S rDNA) sequence showed that RC2-3 belonged to the family Flavobacteriaceae. However, it showed dif-ferent physiological and biochemical characteristics from the known Flavobacteriaceae members producing fucoidan-degrading enzyme, thus RC2-3 was proposed to be a new member of this family.

  8. Isolation and characterization of a fucoidan-degrading bacterium from Laminaria japonica

    Science.gov (United States)

    Wang, Ying; Li, Bafang; Zhao, Xue; Piao, Meizi

    2013-11-01

    Fucoidan, a polysaccharide containing abundant fucose and sulfate ester group, was prepared from Laminaria japonica. In order to obtain fucoidan-degrading enzyme, bacteria capable of degrading fucoidan were screened from kelp. A bacterial strain named RC2-3 was obtained, which degraded fucoidan by the maximum extent of 54% ± 1.3%, the highest among all bacterial isolates. High-performance size exclusion chromatography (HPSEC) showed that the molecular weight of fucoidan was gradually reduced by RC2-3 with culturing time, suggesting the production of fucoidan-degrading enzyme by RC2-3. Phylogenetic analysis of partial 16S ribosomal RNA gene (16S rDNA) sequence showed that RC2-3 belonged to the family Flavobacteriaceae. However, it showed different physiological and biochemical characteristics from the known Flavobacteriaceae members producing fucoidan-degrading enzyme, thus RC2-3 was proposed to be a new member of this family.

  9. Aadh2p: an Arxula adeninivorans alcohol dehydrogenase involved in the first step of the 1-butanol degradation pathway.

    Science.gov (United States)

    Rauter, Marion; Kasprzak, Jakub; Becker, Karin; Riechen, Jan; Worch, Sebastian; Hartmann, Anja; Mascher, Martin; Scholz, Uwe; Baronian, Kim; Bode, Rüdiger; Schauer, Frieder; Matthias Vorbrodt, H; Kunze, Gotthard

    2016-10-12

    The non-conventional yeast Arxula adeninivorans uses 1-butanol as a carbon source and has recently attracted attention as a promising organism for 1-butanol production. Alcohol dehydrogenases (adhp) are important catalysts in 1-butanol metabolism, but only Aadh1p from Arxula has been characterized. This enzyme is involved in ethanol synthesis but has a low impact on 1-butanol degradation. In this study, we identified and characterized a second adhp from A. adeninivorans (Aadh2p). Compared to Saccharomyces cerevisiae ADHs' (ScAdh) protein sequences it originates from the same ancestral node as ScAdh6p, 7p and 4p. It is also localized in the cytoplasm and uses NAD(H) as cofactor. The enzyme has its highest activity with medium chain-length alcohols and maximum activity with 1-butanol with the catalytic efficiency of the purified enzyme being 42 and 43,000 times higher than with ethanol and acetaldehyde, respectively. Arxula adeninivorans strain G1212/YRC102-AADH2, which expresses the AADH2 gene under the control of the strong constitutive TEF1 promoter was constructed. It achieved an ADH activity of up to 8000 U/L and 500 U/g dry cell weight (dcw) which is in contrast to the control strain G1212/YRC102 which had an ADH activity of up to 1400 U/L and 200 U/g dcw. Gene expression analysis showed that AADH2 derepression or induction using non-fermentable carbon-sources such as ethanol, pyruvate, glycerol or 1-butanol did occur. Compared to G1212/YRC102 AADH2 knock-out strain had a slower growth rate and lower 1-butanol consumption if 1-butanol was used as sole carbon source and AADH2-transformants did not grow at all in the same conditions. However, addition of the branched-chain amino acids leucine, isoleucine and valine allowed the transformants to use 1-butanol as carbon source. The addition of these amino acids to the control strain and Δaadh2 mutant cultures had the effect of accelerating 1-butanol consumption. Our results confirm that Aadh2p plays a major

  10. GRAPE: a pathway template method to characterize tissue-specific functionality from gene expression profiles.

    Science.gov (United States)

    Klein, Michael I; Stern, David F; Zhao, Hongyu

    2017-06-26

    Personalizing treatment regimes based on gene expression profiles of individual tumors will facilitate management of cancer. Although many methods have been developed to identify pathways perturbed in tumors, the results are often not generalizable across independent datasets due to the presence of platform/batch effects. There is a need to develop methods that are robust to platform/batch effects and able to identify perturbed pathways in individual samples. We present Gene-Ranking Analysis of Pathway Expression (GRAPE) as a novel method to identify abnormal pathways in individual samples that is robust to platform/batch effects in gene expression profiles generated by multiple platforms. GRAPE first defines a template consisting of an ordered set of pathway genes to characterize the normative state of a pathway based on the relative rankings of gene expression levels across a set of reference samples. This template can be used to assess whether a sample conforms to or deviates from the typical behavior of the reference samples for this pathway. We demonstrate that GRAPE performs well versus existing methods in classifying tissue types within a single dataset, and that GRAPE achieves superior robustness and generalizability across different datasets. A powerful feature of GRAPE is the ability to represent individual gene expression profiles as a vector of pathways scores. We present applications to the analyses of breast cancer subtypes and different colonic diseases. We perform survival analysis of several TCGA subtypes and find that GRAPE pathway scores perform well in comparison to other methods. GRAPE templates offer a novel approach for summarizing the behavior of gene-sets across a collection of gene expression profiles. These templates offer superior robustness across distinct experimental batches compared to existing methods. GRAPE pathway scores enable identification of abnormal gene-set behavior in individual samples using a non-competitive approach that

  11. HUWE1 and TRIP12 Collaborate in Degradation of Ubiquitin-Fusion Proteins and Misframed Ubiquitin

    DEFF Research Database (Denmark)

    Poulsen, Esben G; Steinhauer, Cornelia; Lees, Michael

    2012-01-01

    In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD) pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized. Rec...

  12. Synthesis and characterization of TiO2 pillared montmorillonites: application for methylene blue degradation.

    Science.gov (United States)

    Chen, Daimei; Du, Gaoxiang; Zhu, Qian; Zhou, Fengsan

    2013-11-01

    TiO2 pillared clay composites were prepared by modifying of montmorillonite (Mt) with cetyl-trimethyammoniumbromide (CTAB) and then using an acidic solution of hydrolyzed Ti alkoxide to intercalate into the interlayer space of the organic modified Mt. The as-prepared materials were characterized by XRD, FTIR, TEM, SEM TG-DTA, specific surface area and porosity measurements. The composites had a porous delaminated structure with pillared fragments and well dispersed TiO2 nanoparticles. Introduction of CTAB into the synthetic system accelerated the hydrolysis and condensation of the Ti source, which promoted TiO2 formation. In addition, the CTAB also significantly increased the porosity and surface area of the composites. A number of anatase particles, with crystal sizes of 5-10 nm, were homogenously distributed on the surface of the Mt as the result of the templating role of CTAB. The resultant TiO2 pillared Mt exhibited good thermal stability as indicated by its surface area after calcination at 800°C. No phase transformations from anatase to rutile were observed even under calcination at 900°C. The grain size of the anatase in prepared sample increased from 2.67 nm to 13.42 nm as the calcination temperature increased from 300°C to 900°C. The photocatalytic performance of these new porous materials was evaluated by using methylene blue degradation. The composite exhibited better photocatalytic property than P 25. The maximum removal efficiency of this composite was up to 99% within 60 min.

  13. Characterization and Genomic Analysis of a Highly Efficient Dibutyl Phthalate-Degrading Bacterium Gordonia sp. Strain QH-12

    Science.gov (United States)

    Jin, Decai; Kong, Xiao; Liu, Huijun; Wang, Xinxin; Deng, Ye; Jia, Minghong; Yu, Xiangyang

    2016-01-01

    A bacterial strain QH-12 isolated from activated sludge was identified as Gordonia sp. based on analysis of 16S rRNA gene sequence and was found to be capable of utilizing dibutyl phthalate (DBP) and other common phthalate esters (PAEs) as the sole carbon and energy source. The degradation kinetics of DBP under different concentrations by the strain QH-12 fit well with the modified Gompertz model (R2 > 0.98). However, strain QH-12 could not utilize the major intermediate product phthalate (phthalic acid; PA) as the sole carbon and energy source, and only a little amount of PA was detected. The QH-12 genome analysis revealed the presence of putative hydrolase/esterase genes involved in PAEs-degradation but no phthalic acid catabolic gene cluster was found, suggesting that a novel degradation pathway of PAEs was present in Gordonia sp. QH-12. This information will be valuable for obtaining a more holistic understanding on diverse genetic mechanisms of PAEs-degrading Gordonia sp. strains. PMID:27347943

  14. Characterization and Genomic Analysis of a Highly Efficient Dibutyl Phthalate-Degrading Bacterium Gordonia sp. Strain QH-12

    Directory of Open Access Journals (Sweden)

    Decai Jin

    2016-06-01

    Full Text Available A bacterial strain QH-12 isolated from activated sludge was identified as Gordonia sp. based on analysis of 16S rRNA gene sequence and was found to be capable of utilizing dibutyl phthalate (DBP and other common phthalate esters (PAEs as the sole carbon and energy source. The degradation kinetics of DBP under different concentrations by the strain QH-12 fit well with the modified Gompertz model (R2 > 0.98. However, strain QH-12 could not utilize the major intermediate product phthalate (phthalic acid; PA as the sole carbon and energy source, and only a little amount of PA was detected. The QH-12 genome analysis revealed the presence of putative hydrolase/esterase genes involved in PAEs-degradation but no phthalic acid catabolic gene cluster was found, suggesting that a novel degradation pathway of PAEs was present in Gordonia sp. QH-12. This information will be valuable for obtaining a more holistic understanding on diverse genetic mechanisms of PAEs-degrading Gordonia sp. strains.

  15. Isolation of a diphenylamine-degrading bacterium and characterization of its metabolic capacities, bioremediation and bioaugmentation potential.

    Science.gov (United States)

    Perruchon, Chiara; Batianis, Christos; Zouborlis, Stelios; Papadopoulou, Evangelia S; Ntougias, Spyridon; Vasileiadis, Sotirios; Karpouzas, Dimitrios G

    2015-12-01

    The antioxidant diphenylamine (DPA) is used in fruit-packaging plants for the control of the physiological disorder apple scald. Its use results in the production of DPA-contaminated wastewater which should be treated before finally discharged. Biological treatment systems using tailored-made microbial inocula with specific catabolic activities comprise an appealing and sustainable solution. This study aimed to isolate DPA-degrading bacteria, identify the metabolic pathway of DPA and evaluate their potential for future implementation in bioremediation and biodepuration applications. A Pseudomonas putida strain named DPA1 able to rapidly degrade and utilize DPA as the sole C and N source was enriched from a DPA-contaminated soil. The isolated strain degraded spillage-level concentrations of DPA in liquid culture (2000 mg L(-1)) and in contaminated soil (1000 mg kg(-1)) and metabolized DPA via the transient formation of aniline and catechol. Further evidence for the bioremediation and biodepuration potential of the P. putida strain DPA1 was provided by its capacity to degrade the post-harvest fungicide ortho-phenylphenol (OPP), concurrently used by the fruit-packaging plants, although at slower rates and DPA in a wide range of pH (4.5-9) and temperatures (15-37 °C). These findings revealed the high potential of the P. putida strain DPA1 for use in future soil bioremediation strategies and/or as start-up inocula in wastewater biodepuration systems.

  16. α-Amino acid containing degradable polymers as functional biomaterials: rational design, synthetic pathway, and biomedical applications.

    Science.gov (United States)

    Sun, Huanli; Meng, Fenghua; Dias, Aylvin A; Hendriks, Marc; Feijen, Jan; Zhong, Zhiyuan

    2011-06-13

    Currently, biomedical engineering is rapidly expanding, especially in the areas of drug delivery, gene transfer, tissue engineering, and regenerative medicine. A prerequisite for further development is the design and synthesis of novel multifunctional biomaterials that are biocompatible and biologically active, are biodegradable with a controlled degradation rate, and have tunable mechanical properties. In the past decades, different types of α-amino acid-containing degradable polymers have been actively developed with the aim to obtain biomimicking functional biomaterials. The use of α-amino acids as building units for degradable polymers may offer several advantages: (i) imparting chemical functionality, such as hydroxyl, amine, carboxyl, and thiol groups, which not only results in improved hydrophilicity and possible interactions with proteins and genes, but also facilitates further modification with bioactive molecules (e.g., drugs or biological cues); (ii) possibly improving materials biological properties, including cell-materials interactions (e.g., cell adhesion, migration) and degradability; (iii) enhancing thermal and mechanical properties; and (iv) providing metabolizable building units/blocks. In this paper, recent developments in the field of α-amino acid-containing degradable polymers are reviewed. First, synthetic approaches to prepare α-amino acid-containing degradable polymers will be discussed. Subsequently, the biomedical applications of these polymers in areas such as drug delivery, gene delivery and tissue engineering will be reviewed. Finally, the future perspectives of α-amino acid-containing degradable polymers will be evaluated.

  17. Degradation of ciprofloxacin by 280 nm ultraviolet-activated persulfate: Degradation pathway and intermediate impact on proteome of Escherichia coli.

    Science.gov (United States)

    Ye, Jin-Shao; Liu, Juan; Ou, Hua-Se; Wang, Lin-Lin

    2016-12-01

    In this study, the degradation of ciprofloxacin (CIP) was explored using ultraviolet activated persulfate (UV/PS) with 280 nm ultraviolet light-emitting diodes (UV-LEDs), and the toxicological assessment of degrading intermediates was performed using iTRAQ labeling quantitative proteomic technology. The quantitative mass spectrum results showed that 280 nm UV/PS treatment had a high transformation efficiency of CIP ([CIP] = 3 μM, [S2O8(2-)] = 210 μM, apparent rate constants 0.2413 min(-1)). The high resolution mass spectrum analyses demonstrated that the primary intermediates included C15H16FN3O3 (m/z 306.1248) and C17H18FN3O4 (m/z 348.1354). The former one was formed by the cleavage of piperazine ring, while the later one was generated by the addition of a hydroxyl on the quinolone backbone. The toxicological assessment demonstrated that 56 and 110 proteins had significant up regulations and down regulations, respectively, in the Escherichia coli exposed to degraded CIP compared to untreated CIP. The majority of up-regulated proteins, such as GapA, SodC, were associated with primary metabolic process rather than responses to stress and toxic substance, inferring that the moderate UV/PS treatment can reduce the antibacterial activity of CIP by incomplete mineralization. Consequently, these results provided a novel insight into the application of UV-LED/PS treatment as a promising removal methodology for quinolones. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Biodegradation of 4-nitrotoluene with biosurfactant production by Rhodococcus pyridinivorans NT2: metabolic pathway, cell surface properties and toxicological characterization.

    Science.gov (United States)

    Kundu, Debasree; Hazra, Chinmay; Dandi, Navin; Chaudhari, Ambalal

    2013-11-01

    A novel 4-nitrotoluene-degrading bacterial strain was isolated from pesticides contaminated effluent-sediment and identified as Rhodococcus pyridinivorans NT2 based on morphological and biochemical properties and 16S rDNA sequencing. The strain NT2 degraded 4-NT (400 mg l(-1)) with rapid growth at the end of 120 h, reduced surface tension of the media from 71 to 29 mN m(-1) and produced glycolipidic biosurfactants (45 mg l(-1)). The biosurfactant was purified and characterized as trehalose lipids. The biosurfactant was stable in high salinity (10 % w/v NaCl), elevated temperatures (120 °C for 15 min) and a wide pH range (2.0-10.0). The noticeable changes during biodegradation were decreased hydrophobicity; an increase in degree of fatty acid saturation, saturated/unsaturated ratio and cyclopropane fatty acid. Biodegradation of 4-NT was accompanied by the accumulation of ammonium (NH4 (+)) and negligible amount of nitrite ion (NO2 (-)). Product stoichiometry showed a carbon (C) and nitrogen (N) mass balance of 37 and 35 %, respectively. Biodegradation of 4-NT proceeded by oxidation at the methyl group to form 4-nitrobenzoate, followed by reduction and hydrolytic deamination yielding protocatechuate, which was metabolized through β-ketoadipate pathway. In vitro and in vivo acute toxicity assays in adult rat (Rattus norvegicus) showed sequential detoxification and the order of toxicity was 4-NT >4-nitrobenzyl alcohol >4-nitrobenzaldehyde >4-nitrobenzoate > protocatechuate. Taken together, the strain NT2 could be used as a potential bioaugmentation candidate for the bioremediation of contaminated sites.

  19. Separating and characterizing functional alkane degraders from crude-oil-contaminated sites via magnetic nanoparticle-mediated isolation.

    Science.gov (United States)

    Wang, Xinzi; Zhao, Xiaohui; Li, Hanbing; Jia, Jianli; Liu, Yueqiao; Ejenavi, Odafe; Ding, Aizhong; Sun, Yujiao; Zhang, Dayi

    Uncultivable microorganisms account for over 99% of all species on the planet, but their functions are yet not well characterized. Though many cultivable degraders for n-alkanes have been intensively investigated, the roles of functional n-alkane degraders remain hidden in the natural environment. This study introduces the novel magnetic nanoparticle-mediated isolation (MMI) technology in Nigerian soils and successfully separates functional microbes belonging to the families Oxalobacteraceae and Moraxellaceae, which are dominant and responsible for alkane metabolism in situ. The alkR-type n-alkane monooxygenase genes, instead of alkA- or alkP-type, were the key functional genes involved in the n-alkane degradation process. Further physiological investigation via a BIOLOG PM plate revealed some carbon (Tween 20, Tween 40 and Tween 80) and nitrogen (tyramine, l-glutamine and d-aspartic acid) sources promoting microbial respiration and n-alkane degradation. With further addition of promoter carbon or nitrogen sources, the separated functional alkane degraders significantly improved n-alkane biodegradation rates. This suggests that MMI is a promising technology for separating functional microbes from complex microbiota, with deeper insight into their ecological functions and influencing factors. The technique also broadens the application of the BIOLOG PM plate for physiological research on functional yet uncultivable microorganisms.

  20. Isolation and characterization of a Sphingomonas sp. strain F-7 degrading fenvalerate and its use in bioremediation of contaminated soil.

    Science.gov (United States)

    Yu, Fang B; Shan, Sheng D; Luo, Lin P; Guan, Li B; Qin, Hua

    2013-01-01

    A fenvalerate-degrading bacterial strain F-7 was isolated from long-term contaminated sludge. Based on morphological, physiological and biochemical characterization, and phylogenetic analysis of 16S rRNA gene sequence, strain F-7 was identified as Sphingomonas sp. The bacterium could utilize fenvalerate as the sole source of carbon. An amount measuring 100 mg L(-1) fenvalerate was completely degraded within 72 h and 3-phenoxybenzoic acid (3-PBA) was detected as a major metabolite. The result indicates that S. sp. F-7 might metabolize fenvalerate by hydrolysis of carboxylester linkage. It was capable of degrading permethrin, fenpropathrin, beta-cypermethrin, cyhalothrin, deltamethrin, bifenthrin and 3-PBA. Further studies demonstrated that the strain was multi-resistant to heavy metals and antibiotics. In addition, degradative enzymes involved were confirmed as intracellular distributed and constitutively expressed. Furthermore, application of the strain was found to accelerate the removal of fenvalerate in soil. This is the first report of fenvalerate degrading strain isolated from S. sp. These results might help with future research in better understanding of pyrethroid biodegradation and highlight S. sp. F-7 might have potential for practical application in bioremediation of fenvalerate-contaminated sites.

  1. Isolation and characterization of resin acid degrading bacteria found in effluent from a bleached kraft pulp mill.

    Science.gov (United States)

    Morgan, C A; Wyndham, R C

    1996-05-01

    Thirteen resin acid degrading bacteria enriched on abietic or dehydroabietic acids were isolated from waste water from the aerated stabilization basin of a bleached kraft pulp mill. Standard biochemical tests were used to characterize each isolate. Each isolate was tested for its ability to degrade six abietane- and pimarane-type resin acids. Resin acid concentrations were determined by high pressure liquid chromatography and UV absorbance. Cluster analysis based on phenotypic characteristics identified two distinct clusters of degraders that differed in their ability to utilize carbohydrates as carbon sources. Fatty acid methyl ester analysis of representative isolates from each cluster identified A19-6a and D11-13 as Comamonas and Alcaligenes species, respectively. To determine genotypic relatedness, enterobacterial repetitive intergenic consensus sequences were used to amplify genomic DNA fragments from 10 isolates. These results supported the phenotypic analysis for all isolates tested except A19-5 and A19-6b. These two organisms were clustered closely together based on phenotype but had distinctly different banding patterns, suggesting that they are not related genotypically. All isolates degraded a subset of the six resin acid congeners. Isolates A19-3, A19-6a, A19-6b, and D11-37 were the most effective at degrading all six congeners.

  2. Fucoidan inhibition of lung cancer in vivo and in vitro : role of the Smurf2-dependent ubiquitin proteasome pathway in TGFβ receptor degradation.

    Science.gov (United States)

    Hsu, Hsien-Yeh; Lin, Tung-Yi; Wu, Yu-Chung; Tsao, Shu-Ming; Hwang, Pai-An; Shih, Yu-Wei; Hsu, Jason

    2014-09-15

    Fucoidan, a polysaccharide extracted from brown seaweeds, reduces tumor cell proliferation. In this study, we demonstrate that fucoidan reduces tumor size in LLC1-xenograft male C57BL/6 mice. Moreover, we found that LLC1-bearing mice continuously fed fucoidan showed greater antitumor activity than mice with discontinuous feeding. Fucoidan inhibited the in vitro growth of lung cancer cells. Transforming growth factor β (TGFβ) receptors (TGFRs) play important roles in the regulation of proliferation and progression, and high TGFRI expression in lung cancer specimens is associated with a worse prognosis. Herein, using lung cancer cells, we found that fucoidan effectively reduces TGFRI and TGFRII protein levels in vivo and in vitro. Moreover, fucoidan reduces TGFR downstream signaling events, including those in Smad2/3 and non-Smad pathways: Akt, Erk1/2, and FAK phosphorylation. Furthermore, fucoidan suppresses lung cancer cell mobility upon TGFβ stimulation. To elucidate how fucoidan decreases TGFR proteins in lung cancer cells, we found that fucoidan enhances the ubiquitination proteasome pathway (UPP)-mediated degradation of TGFRs in A549 and CL1-5 cells. Mechanistically, fucoidan promotes Smurf2 and Smad7 to conjugate TGFRs, resulting in TGF degradation; however, Smurf2-shRNA abolishes fucoidan-enhanced UPP-mediated TGFR degradation. Our study is the first to identify a novel mechanism for the antitumor activity of fucoidan, namely decreasing tumor growth by modulating the TGFR/Smad7/Smurf2-dependent axis, leading to TGFR protein degradation and inhibition of lung cancer cell progression in vitro and in vivo. Our current findings indicate that fucoidan is a potential therapeutic agent or dietary supplementation for lung cancer, acting via the Smurf2-dependent ubiquitin degradation of TGFβ receptors.

  3. Reaction pathways of dimethyl phthalate degradation in TiO2-UV-O2 and TiO2-UV-Fe(VI) systems.

    Science.gov (United States)

    Yuan, Bao-ling; Li, Xiang-zhong; Graham, Nigel

    2008-05-01

    The photocatalytic degradation of dimethyl phthalate (DMP) in aqueous TiO2 suspension under UV illumination has been investigated using oxygen (O2) and ferrate (Fe(VI)) as electron acceptors. The experiments demonstrated that Fe(VI) was a more effective electron acceptor than O2 for scavenging the conduction band electrons from the surface of the catalyst. Some major intermediate products from DMP degradation were identified by HPLC and GC/MS analyses. The analytical results identified dimethyl 3-hydroxyphthalate and dimethyl 2-hydroxyphthalate as the two main intermediate products from the DMP degradation in the TiO2-UV-O2 system, while in contrast phthalic acid was found to be the main intermediate product in the TiO2-UV-Fe(VI) system. These findings indicate that DMP degradation in the TiO2-UV-O2 and TiO2-UV-Fe(VI) systems followed different reaction pathways. An electron spin resonance analysis confirmed that hydroxyl radicals existed in the TiO2-UV-O2 reaction system and an unknown radical species (most likely an iron-oxo species) is suspected to exist in the TiO2-UV-Fe(VI) reaction system. Two pathway schemes of DMP degradation in the TiO2-UV-O2 and TiO2-UV-Fe(VI) reaction systems are proposed. It is believed that the radicals formed in the TiO2-UV-O2 reaction system preferably attack the aromatic ring of the DMP, while in contrast the radicals formed in the TiO2-UV-Fe(VI) reaction systems attack the alkyl chain of DMP.

  4. Microwave-assisted polyol synthesis and characterization of pvp-capped cds nanoparticles for the photocatalytic degradation of tartrazine

    Energy Technology Data Exchange (ETDEWEB)

    Darwish, Maher, E-mail: m-darwish@razi.tums.ac.ir [Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Mohammadi, Ali, E-mail: alimohammadi@tums.ac.ir [Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Assi, Navid, E-mail: navid_a30@yahoo.com [Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2016-02-15

    Highlights: • PVP-stabilized CdS nanoparticles have been fabricated by a polyol-microwave method. • CdS nanoparticles were characterized and the size was approximately 48 ± 10 nm. • Catalytic activity of our nanoparticles was examined for tartrazine degradation. • Remarkable results were obtained under both UV and visible light irradiations. - Abstract: Polyvinylpyrrolidone capped cadmium sulfide nanoparticles have been successfully synthesized by a facile polyol method with ethylene glycol. Microwave irradiation and calcination were used to control the size and shape of nanoparticles. Characterization with scanning electron microscopy revealed a restricted nanoparticles growth comparing with the uncapped product, hexagonal phase and 48 nm average particle size were confirmed by X-ray diffraction, and finally mechanism of passivation was suggested depending on Fourier transform infrared spectra. The efficiency of nanoparticles was evaluated by the photocatalytic degradation of tartrazine in aqueous solution under UVC and visible light irradiation. Complete degradation of the dye was observed after 90 min of UVC irradiation under optimized conditions. Kinetic of reaction fitted well to the pseudo-first-order kinetic and Langmuir–Hinshelwood models. Furthermore, 85% degradation of the dye in 9 h under visible light suggests that cadmium sulfide is a promising tool to work under visible light for environmental remediation.

  5. Characterization and prevention of humidity related degradation of atomic layer deposited Al2O3

    Science.gov (United States)

    Rückerl, Andreas; Zeisel, Roland; Mandl, Martin; Costina, Ioan; Schroeder, Thomas; Zoellner, Marvin H.

    2017-01-01

    Atomic layer deposited aluminum oxide (ALD-Al2O3) is a dielectric material, which is widely used in organic light emitting diodes in order to prevent their organic layers from humidity related degradation. Unfortunately, there are strong hints that in some cases, ALD-Al2O3 itself is suffering from humidity related degradation. Especially, high temperature and high humidity seem to enhance ALD-Al2O3 degradation strongly. For this reason, the degradation behavior of ALD-Al2O3 films at high temperature and high humidity was investigated in detail and a way to prevent it from degradation was searched. The degradation behavior is analyzed in the first part of this paper. Using infrared absorbance measurements and X-ray diffraction, boehmite (γ-AlOOH) was identified as a degradation product. In the second part of the paper, it is shown that ALD-Al2O3 films can be effectively protected from degradation using a silicon oxide capping. The deposition of very small amounts of silicon in a molecular beam epitaxy system and an X-ray photoelectron spectroscopy investigation of the chemical bonding between the silicon and the ALD-Al2O3 surface led to the conclusion that a silicon termination of the ALD-Al2O3 surface (Al*-O-SiOx) is able to stop humidity related degradation of the underlying ALD-Al2O3 films. The third part of the paper shows that the protection mechanism of the silicon termination is probably due to the strong tendency of silicic acid to resilificate exposed ALD-Al2O3 surfaces. The protective effect of a simple silicon source on an ALD-Al2O3 surface is shown exemplary and the related chemical reactions are presented.

  6. Enrichment and Characterization of PCB-Degrading Bacteria as Potential Seed Cultures for Bioremediation of Contaminated Soil

    Directory of Open Access Journals (Sweden)

    Dubravka Hršak

    2007-01-01

    Full Text Available The main objective of our study was to obtain seed cultures for enhancing the transformation of polychlorinated biphenyls (PCBs in contaminated soil of the transformer station in Zadar, Croatia, damaged during warfare activities in 1991. For enrichment, six soil samples were collected from different polluted areas and microcosm approach, stimulating the growth of biphenyl-degrading bacteria, was employed. Enrichment experiments resulted in the selection of two fast growing mixed cultures TSZ7 and AIR1, originating from the soil of the transformer station and the airport area, respectively. Both cultures showed significant PCB-degrading activity (56 to 60 % of PCB50 mixture was reduced after a two-week cultivation. Furthermore, the cultures displayed similar PCB-degrading competence and reduced di-to tetrachlorobiphenyls more effectively than penta- to hepta-chlorobiphenyls. Strain Z6, identified as Rhodococcus erythropolis, was found to be the only culture member showing PCB-transformation potential similar to that of the mixed culture TSZ7, from which it was isolated. Based on the metabolites identified in the assay with the single congener 2,4,4’-chlorobiphenyl, we proposed that the strain Z6 was able to use both the 2,3-and 3,4-dioxygenase pathways. Furthermore, the identified metabolites suggested that beside these pathways another unidentified pathway might also be active in strain Z6. Based on the obtained results, the culture TSZ7 and the strain Z6 were designated as potential seed cultures for bioremediation of the contaminated soil.

  7. Ti/ZnO-MxOy composites (M = Al, Cr, Fe, Ce): synthesis, characterization and application as highly efficient photocatalysts for hexachlorobenzene degradation.

    Science.gov (United States)

    Xia, Shengjie; Shao, Mengmeng; Zhou, Xiaobo; Pan, Guoxiang; Ni, Zheming

    2015-10-28

    A series of novel organic-inorganic nanoscale layered materials were synthesized by intercalating the Ti-containing Schiff base complex into the interlayer of the ZnM layered double hydroxides (LDHs, M = Al, Cr, Fe, Ce). The hybrid material was further calcined to make metal oxide composites with highly dispersed Ti elements (Ti/ZnO-MxOy). The structural characterization and photocatalytic results showed that, after intercalation and calcination, the metal oxide composites with a unique flower-like crystal morphology not only had high specific surface area, uniform pore size distribution and narrow band gap, but also showed extremely high photocatalytic performance for hexachlorobenzene (HCB) degradation. The Ti/ZnO-Cr2O3 composite with the narrowest band gap (2.40 eV) and the highest surface area (227 m(2)) showed the highest photocatalytic performance for HCB (95.5% within 240 min) among the four metal oxide composites. Particularly, it was found that composites derived from layered materials with different supramolecular structure of the host and guest showed different photocatalytic properties. In addition, based on the results from ESR, GC-MS and HPLC-MS, the type and amount of hydroxyl radicals, the decomposition intermediates and the pathway of HCB degradation photocatalyzed by Ti/ZnO-MxOy composites are also discussed in detail.

  8. Isolation and characterization of a bacterium that degrades various polyester-based biodegradable plastics.

    Science.gov (United States)

    Teeraphatpornchai, T; Nakajima-Kambe, T; Shigeno-Akutsu, Y; Nakayama, M; Nomura, N; Nakahara, T; Uchiyama, H

    2003-01-01

    Microorganisms isolated from soil samples were screened for their ability to degrade various biodegradable polyester-based plastics. The most active strain, designated as strain TB-13, was selected as the best strain for degrading these plastics. From its phenotypic and genetic characteristics, strain TB-13 was closely related to Paenibacillus amyloyticus. It could degrade poly(lactic acid), poly(butylene succinate), poly(butylene succinate-co-adipate), poly(caprolactone) and poly(ethylene succinate) but not poly(hydroxybutylate-co-valerate). However, it could not utilize these plastics as sole carbon sources. Both protease and esterase activities, which may be involved in the degradation of plastic, were constitutively detected in the culture broth.

  9. Isolation and characterization of gasoline-degrading bacteria from gas station leaking-contaminated soils

    Institute of Scientific and Technical Information of China (English)

    LU Si-jin; WANG Hong-qi; YAO Zhi-hua

    2006-01-01

    The effects of culture conditions in vitro and biosurfactant detection were studied on bacterial strains capable of degrading gasoline from contaminated soils near gas station. The main results were summarized as follows. Three bacteria (strains Q10, Q14 and Q18) that were considered as efficiently degrading strains were isolated and identified as Pseudomonas sp., Flavobacterium sp. and Rhodococcus sp., respectively. The optimal growth conditions of three bacteria including pH, temperature and the concentration of gasoline were similar. The reduction in surface tension was observed with all the three bacteria, indicating the production of toluene, ethylbenzene and xylene (BTEX) could easily be degraded by the three isolates. The consortium was more effective than the individual cultures in degrading added gasoline, diesel oil, and BTEX. These results indicate that these strains have great potential for in situ remediation of soils contaminated by gas station leaking.

  10. Application of {sup 13}C and {sup 15}N stable isotope probing to characterize RDX degrading microbial communities under different electron-accepting conditions

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Kun-Ching; Lee, Do Gyun [Zachry Department of Civil Engineering, Texas A& M University, College Station, TX 77843-3136 (United States); Fuller, Mark E.; Hatzinger, Paul B.; Condee, Charles W. [CB& I Federal Services, Lawrenceville, NJ (United States); Chu, Kung-Hui, E-mail: kchu@civil.tamu.edu [Zachry Department of Civil Engineering, Texas A& M University, College Station, TX 77843-3136 (United States)

    2015-10-30

    Highlights: • SIP characterized RDX-degrading communities under different e-accepting conditions. • Dominant RDX degradation pathways differed under different e-accepting conditions. • More complete detoxification of RDX occurred under methanogenic and sulfate-reducing conditions than under manganese(IV) and iron(III)-reducing conditions. - Abstract: This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or its metabolites as carbon and/or nitrogen sources under different electron-accepting conditions using {sup 13}C and {sup 15}N stable isotope probing (SIP). Mesocosms were constructed using groundwater and aquifer solids from an RDX-contaminated aquifer. The mesocosms received succinate as a carbon source and one of four electron acceptors (nitrate, manganese(IV), iron(III), or sulfate) or no additional electron acceptor (to stimulate methanogenesis). When RDX degradation was observed, subsamples from each mesocosm were removed and amended with {sup 13}C{sub 3}- or ring-{sup 15}N{sub 3}-, nitro-{sup 15}N{sub 3}-, or fully-labeled {sup 15}N{sub 6}-RDX, followed by additional incubation and isolation of labeled nucleic acids. A total of fifteen 16S rRNA sequences, clustering in α- and γ-Proteobacteria, Clostridia, and Actinobacteria, were detected in the {sup 13}C-DNA fractions. A total of twenty seven sequences were derived from different {sup 15}N-DNA fractions, with the sequences clustered in α- and γ-Proteobacteria, and Clostridia. Interestingly, sequences identified as Desulfosporosinus sp. (in the Clostridia) were not only observed to incorporate the labeled {sup 13}C or {sup 15}N from labeled RDX, but also were detected under each of the different electron-accepting conditions. The data suggest that {sup 13}C- and {sup 15}N-SIP can be used to characterize microbial communities involved in RDX biodegradation, and that the dominant pathway of RDX biodegradation may differ under different

  11. Characterizing peat palm forest degradation in the Peruvian Amazon from space and on the ground

    Science.gov (United States)

    Hergoualc'h, Kristell; Gutierrez-Velez, Victor Hugo; van Lent, Jeffrey; Verchot, Louis Vincent

    2016-04-01

    Peru has the second largest area of peatlands in the Tropics however little is known on how the biogeochemical cycle of its peat forests can be affected through anthropogenic intervention. The most representative land cover on peat is a Mauritia flexuosa-dominated palm swamp forest which has been under human pressure over decades due the high demand for the M. flexuosa fruit often collected by cutting down the entire palm. Degradation of these carbon-dense forests can severely affect emissions of greenhouse gases and contribute to climate change. The objectives of this research were to assess the impacts on soil trace gas fluxes and biomass carbon stocks of peat palm swamp forest degradation and to explore the potential of remote sensing methods combined with field measurements to map the distribution of peat palm swamp forest according degradation levels. Results suggest a shift in forest composition from palm- to woody-tree dominated forest following degradation. We also found that human intervention in peat palm swamp forest can translate into substantial reductions in tree carbon stocks with a decrease in initial biomass (above and below-ground) stocks (118.3 ± 1.1 Mg C ha-1) by 26 and 44% following medium and high degradation. Preliminary results suggest high and low soil CH4 and CO2 emission rates on average, as compared to Southeast Asian peat swamp forests whereas N2O emissions are of the same magnitude. Degradation seems to disrupt soil respiration mainly through micro-climatic changes induced by reduced canopy cover. The analysis indicates a good potential to discriminate areas of peat palm swamp forest with different levels of degradation from other land covers, suggesting the feasibility of monitoring peat palm swamp forest degradation using remote sensing analyses.

  12. Isolation and characterization of heavy polycyclic aromatic hydrocarbon-degrading bacteria adapted to electrokinetic conditions.

    Science.gov (United States)

    Li, Fengmei; Guo, Shuhai; Hartog, Niels; Yuan, Ye; Yang, Xuelian

    2016-02-01

    Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria capable of growing under electrokinetic conditions were isolated using an adjusted acclimation and enrichment procedure based on soil contaminated with heavy PAHs in the presence of an electric field. Their ability to degrade heavy PAHs under an electric field was individually investigated in artificially contaminated soils. The results showed that strains PB4 (Pseudomonas fluorescens) and FB6 (Kocuria sp.) were the most efficient heavy PAH degraders under electrokinetic conditions. They were re-inoculated into a polluted soil from an industrial site with a PAH concentration of 184.95 mg kg(-1). Compared to the experiments without an electric field, the degradation capability of Pseudomonas fluorescens and Kocuria sp. was enhanced in the industrially polluted soil under electrokinetic conditions. The degradation extents of total PAHs were increased by 15.4 and 14.0% in the electrokinetic PB4 and FB6 experiments (PB4 + EK and FB6 + EK) relative to the PB4 and FB6 experiments without electrokinetic conditions (PB4 and FB6), respectively. These results indicated that P. fluorescens and Kocuria sp. could efficiently degrade heavy PAHs under electrokinetic conditions and have the potential to be used for the electro-bioremediation of PAH-contaminated soil, especially if the soil is contaminated with heavy PAHs.

  13. Comparative functional characterization of the CSR-1 22G-RNA pathway in Caenorhabditis nematodes.

    Science.gov (United States)

    Tu, Shikui; Wu, Monica Z; Wang, Jie; Cutter, Asher D; Weng, Zhiping; Claycomb, Julie M

    2015-01-01

    As a champion of small RNA research for two decades, Caenorhabditis elegans has revealed the essential Argonaute CSR-1 to play key nuclear roles in modulating chromatin, chromosome segregation and germline gene expression via 22G-small RNAs. Despite CSR-1 being preserved among diverse nematodes, the conservation and divergence in function of the targets of small RNA pathways remains poorly resolved. Here we apply comparative functional genomic analysis between C. elegans and Caenorhabditis briggsae to characterize the CSR-1 pathway, its targets and their evolution. C. briggsae CSR-1-associated small RNAs that we identified by immunoprecipitation-small RNA sequencing overlap with 22G-RNAs depleted in cbr-csr-1 RNAi-treated worms. By comparing 22G-RNAs and target genes between species, we defined a set of CSR-1 target genes with conserved germline expression, enrichment in operons and more slowly evolving coding sequences than other genes, along with a small group of evolutionarily labile targets. We demonstrate that the association of CSR-1 with chromatin is preserved, and show that depletion of cbr-csr-1 leads to chromosome segregation defects and embryonic lethality. This first comparative characterization of a small RNA pathway in Caenorhabditis establishes a conserved nuclear role for CSR-1 and highlights its key role in germline gene regulation across multiple animal species.

  14. Phosphate Shifted Oxygen Reduction Pathway on Fe@Fe2O3 Core-Shell Nanowires for Enhanced Reactive Oxygen Species Generation and Aerobic 4-Chlorophenol Degradation.

    Science.gov (United States)

    Mu, Yi; Ai, Zhihui; Zhang, Lizhi

    2017-07-18

    Phosphate ions widely exist in the environment. Previous studies revealed that the adsorption of phosphate ions on nanoscale zerovalent iron would generate a passivating oxide shell to block reactive sites and thus decrease the direct pollutant reduction reactivity of zerovalent iron. Given that molecular oxygen activation process is different from direct pollutant reduction with nanoscale zerovalent iron, it is still unclear how phosphate ions will affect molecular oxygen activation and reactive oxygen species generation with nanoscale zerovalent iron. In this study, we systematically studied the effect of phosphate ions on molecular oxygen activation with Fe@Fe2O3 nanowires, a special nanoscale zerovalent iron, taking advantages of rotating ring disk electrochemical analysis. It was interesting to find that the oxygen reduction pathway on Fe@Fe2O3 nanowires was gradually shifted from a four-electron reduction pathway to a sequential one-electron reduction one, along with increasing the phosphate ions concentration from 0 to 10 mmol·L(-1). This oxygen reduction pathway change greatly enhanced the molecular oxygen activation and reactive oxygen species generation performances of Fe@Fe2O3 nanowires, and thus increased their aerobic 4-chlorophenol degradation rate by 10 times. These findings shed insight into the possible roles of widely existed phosphate ions in molecular oxygen activation and organic pollutants degradation with nanoscale zerovalent iron.

  15. Non-degradative ubiquitination of the Notch1 receptor by the E3 ligase MDM2 activates the Notch signalling pathway.

    Science.gov (United States)

    Pettersson, Susanne; Sczaniecka, Matylda; McLaren, Lorna; Russell, Fiona; Gladstone, Karen; Hupp, Ted; Wallace, Maura

    2013-03-15

    The Notch receptor is necessary for modulating cell fate decisions throughout development, and aberrant activation of Notch signalling has been associated with many diseases, including tumorigenesis. The E3 ligase MDM2 (murine double minute 2) plays a role in regulating the Notch signalling pathway through its interaction with NUMB. In the present study we report that MDM2 can also exert its oncogenic effects on the Notch signalling pathway by directly interacting with the Notch 1 receptor through dual-site binding. This involves both the N-terminal and acidic domains of MDM2 and the RAM [RBP-Jκ (recombination signal-binding protein 1 for Jκ)-associated molecule] and ANK (ankyrin) domains of Notch 1. Although the interaction between Notch1 and MDM2 results in ubiquitination of Notch1, this does not result in degradation of Notch1, but instead leads to activation of the intracellular domain of Notch1. Furthermore, MDM2 can synergize with Notch1 to inhibit apoptosis and promote proliferation. This highlights yet another target for MDM2-mediated ubiquitination that results in activation of the protein rather than degradation and makes MDM2 an attractive target for drug discovery for both the p53 and Notch signalling pathways.

  16. Use of gene expression and pathway signatures to characterize the complexity of human melanoma.

    Science.gov (United States)

    Freedman, Jennifer A; Tyler, Douglas S; Nevins, Joseph R; Augustine, Christina K

    2011-06-01

    A defining characteristic of most human cancers is heterogeneity, resulting from the somatic acquisition of a complex array of genetic and genomic alterations. Dissecting this heterogeneity is critical to developing an understanding of the underlying mechanisms of disease and to paving the way toward personalized treatments of the disease. We used gene expression data sets from the analysis of primary and metastatic melanomas to develop a molecular description of the heterogeneity that characterizes this disease. Unsupervised hierarchical clustering, gene set enrichment analyses, and pathway activity analyses were used to describe the genetic heterogeneity of melanomas. Patterns of gene expression that revealed two distinct classes of primary melanoma, two distinct classes of in-transit melanoma, and at least three subgroups of metastatic melanoma were identified. Expression signatures developed to predict the status of oncogenic signaling pathways were used to explore the biological basis underlying these differential patterns of expression. This analysis of activities revealed unique pathways that distinguished the primary and metastatic subgroups of melanoma. Distinct patterns of gene expression across primary, in-transit, and metastatic melanomas underline the genetic heterogeneity of this disease. This heterogeneity can be described in terms of deregulation of signaling pathways, thus increasing the knowledge of the biological features underlying individual melanomas and potentially directing therapeutic opportunities to individual patients with melanoma.

  17. Removal of 2,4-dichlorophenoxyacetic acid in aqueous solution by pulsed corona discharge treatment: Effect of different water constituents, degradation pathway and toxicity assay.

    Science.gov (United States)

    Singh, Raj Kamal; Philip, Ligy; Ramanujam, Sarathi

    2017-10-01

    A multiple pin-plane corona discharge reactor was used to generate plasma for the degradation of 2,4 dichlorophenoxyacetic acid (2,4-D) from the aqueous solution. The 2,4-D of concentration 1 mg/L was completely removed within 6 min of plasma treatment. Almost complete mineralization was achieved after the treatment time of 14 min for a 2,4-D concentration of 10 mg/L. Effects of different water constituents such as carbonates, nitrate, sulphate, chloride ions, natural organic matter (humic acids) and pH on 2,4-D degradation was studied. A significant antagonistic effect of carbonate and humic acid was observed, whereas, the effects of other ions were insignificant. A higher first order rate constant of 1.73 min(-1) was observed, which was significantly decreased in the presence of carbonate ions and humic acids. Also, a higher degradation of 2,4-D was observed in acidic pH conditions. Different 2,4-D intermediates were detected and the degradation pathway of 2,4-D in plasma treatment process was suggested. The toxicity of 10 mg/L 2,4-D was completely eradicated after 10 min of plasma treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Matrix Metalloproteinase-9 Leads to Claudin-5 Degradation via the NF-κB Pathway in BALB/c Mice with Eosinophilic Meningoencephalitis Caused by Angiostrongylus cantonensis

    Science.gov (United States)

    Chiu, Ping-Sung; Lai, Shih-Chan

    2013-01-01

    The epithelial barrier regulates the movement of ions, macromolecules, immune cells and pathogens. The objective of this study was to investigate the role of the matrix metalloproteinase (MMP)-9 in the degradation of tight junction protein during infection with rat nematode lungworm Angiostrongylus cantonensis. The results showed that phosphorylation of IκB and NF-κB was increased in mice with eosinophilic meningoencephalitis. Treatment with MG132 reduced the phosphorylation of NF-κB and the activity of MMP-9, indicating upregulation of MMP-9 through the NF-κB signaling pathway. Claudin-5 was reduced in the brain but elevated in the cerebrospinal fluid (CSF), implying that A. cantonensis infection caused tight junction breakdown and led to claudin-5 release into the CSF. Degradation of claudin-5 coincided with alteration of the blood-CSF barrier permeability and treatment with the MMP inhibitor GM6001 attenuated the degradation of claudin-5. These results suggested that degradation of claudin-5 was caused by MMP-9 in angiostrongyliasis meningoencephalitis. Claudin-5 could be used for the pathophysiologic evaluation of the blood-CSF barrier breakdown and tight junction disruption after infection with A. cantonensis. PMID:23505411

  19. Common Degradative Pathways of Morpholine, Thiomorpholine, and Piperidine by Mycobacterium aurum MO1: Evidence from 1H-Nuclear Magnetic Resonance and Ionspray Mass Spectrometry Performed Directly on the Incubation Medium

    Science.gov (United States)

    Combourieu, Bruno; Besse, Pascale; Sancelme, Martine; Godin, Jean-Philippe; Monteil, André; Veschambre, Henri; Delort, Anne-Marie

    2000-01-01

    In order to see if the biodegradative pathways for morpholine and thiomorpholine during degradation by Mycobacterium aurum MO1 could be generalized to other heterocyclic compounds, the degradation of piperidine by this strain was investigated by performing 1H-nuclear magnetic resonance directly with the incubation medium. Ionspray mass spectrometry, performed without purification of the samples, was also used to confirm the structure of some metabolites during morpholine and thiomorpholine degradation. The results obtained with these two techniques suggested a general pathway for degradation of nitrogen heterocyclic compounds by M. aurum MO1. The first step of the degradative pathway is cleavage of the C—N bond; this leads formation of an intermediary amino acid, which is followed by deamination and oxidation of this amino acid into a diacid. Except in the case of thiodiglycolate obtained from thiomorpholine degradation, the dicarboxylates are completely mineralized by the bacterial cells. A comparison with previously published data showed that this pathway could be a general pathway for degradation by other strains of members of the genus Mycobacterium. PMID:10919768

  20. GBF1, a transcription factor of blue light signaling in Arabidopsis, is degraded in the dark by a proteasome-mediated pathway independent of COP1 and SPA1.

    Science.gov (United States)

    Mallappa, Chandrashekara; Singh, Aparna; Ram, Hathi; Chattopadhyay, Sudip

    2008-12-19

    Arabidopsis GBF1/ZBF2 is a bZIP transcription factor that plays dual but opposite regulatory roles in cryptochrome-mediated blue light signaling. Here, we show the genetic and molecular interrelation of GBF1 with two well characterized negative regulators of light signaling, COP1 and SPA1, in photomorphogenic growth and light-regulated gene expression. Our results further reveal that GBF1 protein is less abundant in the dark-grown seedlings and is degraded by a proteasome-mediated pathway independent of COP1 and SPA1. Furthermore, COP1 physically interacts with GBF1 and is required for the optimum accumulation of GBF1 protein in light-grown seedlings. Taken together, this study provides a mechanistic view of concerted function of three important regulators in Arabidopsis seedling development.

  1. Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing

    Directory of Open Access Journals (Sweden)

    Vasudeo Zambare, Archana Zambare, Kasiviswanath Muthukumarappan, Lew P. Christopher

    2011-01-01

    Escherichia coli. This could have important implications in the enzymatic breakdown of lignocellulosic biomass for the establishment of a robust and cost-efficient process for production of cellulosic ethanol. To the best of our knowledge, this work represents the first report in literature on biochemical characterization of lignocellulose-degrading enzymes from a thermophilic microbial consortium.

  2. Preparation, characterization and application of nanosized copper ferrite photocatalysts for dye degradation under UV irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Zaharieva, Katerina, E-mail: zaharieva@ic.bas.bg [Institute of Catalysis, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 11, 1113 Sofia (Bulgaria); Rives, Vicente, E-mail: vrives@usal.es [GIR-QUESCAT, Dpto. Química Inorgánica, Universidad de Salamanca, 37008 Salamanca (Spain); Tsvetkov, Martin, E-mail: mptsvetkov@gmail.com [Faculty of Chemistry and Pharmacy, St. Kliment Ohridski University of Sofia, 1 J. Bourchier Blvd., 1164 Sofia (Bulgaria); Cherkezova-Zheleva, Zara, E-mail: zzhel@ic.bas.bg [Institute of Catalysis, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 11, 1113 Sofia (Bulgaria); Kunev, Boris, E-mail: bkunev@ic.bas.bg [Institute of Catalysis, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 11, 1113 Sofia (Bulgaria); Trujillano, Raquel, E-mail: rakel@usal.es [GIR-QUESCAT, Dpto. Química Inorgánica, Universidad de Salamanca, 37008 Salamanca (Spain); Mitov, Ivan, E-mail: mitov@ic.bas.bg [Institute of Catalysis, Bulgarian Academy of Sciences, Acad. G. Bonchev St., Block 11, 1113 Sofia (Bulgaria); Milanova, Maria, E-mail: nhmm@wmail.chem.uni-sofia.bg [Faculty of Chemistry and Pharmacy, St. Kliment Ohridski University of Sofia, 1 J. Bourchier Blvd., 1164 Sofia (Bulgaria)

    2015-06-15

    Nanosized copper ferrite-type materials (Cu{sub x}Fe{sub 3–x}O{sub 4}, 0 ≤ x ≤ 1) have been prepared by combination of co-precipitation and mechanochemical activation and/or thermal treatment. The crystalline structure and morphology of the obtained ferrite nanopowders have been characterized by different instrumental methods, such as Powder X-ray diffraction (PXRD), Mössbauer and FT-IR spectroscopies, specific surface area and porosity measurements, thermal analyses (Differential Thermal Analysis and Thermogravimetric Analysis) and Temperature-Programmed Reduction. The average crystallite size of copper ferrites ranged between 7.8 and 14.7 nm and show a superparamagnetic and collective magnetic excitations nature. The photocatalytic decolorization of Malachite green oxalate under different UV illumination intervals was examined using these copper ferrites as photocatalysts. The results indicate that the prepared nanostructured copper ferrites showed enhanced photocatalytic activity and amount adsorbed Malachite Green dye. The co-precipitated nanosized copper ferrite powder with a low content of copper metal ions in a magnetite host structure (Cu{sub 0.25}Fe{sub 2.75}O{sub 4}) showed an apparent pseudo-first-order rate constant 15.4 × 10{sup −3} min{sup −1} and an amount adsorbed Malachite Green as model organic dye pollutant per 1 g catalyst of 33.4 ppm/g after the dark period. The results confirm that the copper ferrites can be suitable for photocatalytic treatment of wastewaters containing organic dyes. The new aspect of presented investigations is to study the influence of different degree of incorporation of copper ions into the magnetite host structure and preparation methods on the photocatalytic properties of nanosized copper ferrite materials and obtaining of potential photocatalyst (Cu{sub 0.25}Fe{sub 2.75}O{sub 4}) with higher photocatalytic activity (15.4 × 10{sup −3} min{sup −1}) than that of the standard referent Degussa P25 (12 × 10

  3. Functional characterization of p53 pathway components in the ancient metazoan Trichoplax adhaerens

    Science.gov (United States)

    Siau, Jia Wei; Coffill, Cynthia R.; Zhang, Weiyun Villien; Tan, Yaw Sing; Hundt, Juliane; Lane, David; Verma, Chandra; Ghadessy, Farid

    2016-09-01

    The identification of genes encoding a p53 family member and an Mdm2 ortholog in the ancient placozoan Trichoplax adhaerens advocates for the evolutionary conservation of a pivotal stress-response pathway observed in all higher eukaryotes. Here, we recapitulate several key functionalities ascribed to this known interacting protein pair by analysis of the placozoan proteins (Tap53 and TaMdm2) using both in vitro and cellular assays. In addition to interacting with each other, the Tap53 and TaMdm2 proteins are also able to respectively bind human Mdm2 and p53, providing strong evidence for functional conservation. The key p53-degrading function of Mdm2 is also conserved in TaMdm2. Tap53 retained DNA binding associated with p53 transcription activation function. However, it lacked transactivation function in reporter genes assays using a heterologous cell line, suggesting a cofactor incompatibility. Overall, the data supports functional roles for TaMdm2 and Tap53, and further defines the p53 pathway as an evolutionary conserved fulcrum mediating cellular response to stress.

  4. Isolation and Characterization of a Thermophilic Oil-Degrading Bacterial Consortium

    Institute of Scientific and Technical Information of China (English)

    Gu Guizhou; Li Zheng; Zhao Dongfeng; Zhao Chaocheng

    2013-01-01

    In this study, a thermophilic oil-degrading bacterial consortium KO8-2 growing within the temperature range of 45-65℃(with 55℃being the optimum temperature) was isolated from oil-contaminated soil of Karamay in Xinjiang, China. Denaturing gradient gel electrophoresis (DGGE) showed that there were nine strains included in KO8-2, which originated from the genera of Bacillus, Geobacillus and Clostridium. They all belonged to thermophilic bacteria, and had been previously proved as degraders of at least one petroleum fraction. The crude oil degraded by KO8-2 was analyzed by infrared spectrophotometry, hydrocarbon group type analysis and gas chromatography. The results indicated that the bacterial consortium KO8-2 was able to utilize 64.33%of saturates, 27.06%of aromatics, 13.24%of resins and the oil removal efifciency reached up to 58.73%at 55℃when the oil concentration was 10 g/L. Detailed analysis showed that KO8-2 was able to utilize the hydrocarbon components before C19, and the n-alkanes ranging from C20-C33 were signiif-cantly degraded. The ratios of nC17/Pr and nC18/Ph were 3.12 and 3.87, respectively, before degradation, whereas after degradation the ratios reduced to 0.21 and 0.38, respectively. Compared with the control sample, the oil removal efifciency in KO8-2 composting reactor reached 50.12%after a degradation duration of 60 days.

  5. NEDD4 E3 ligase inhibits the activity of the Hippo pathway by targeting LATS1 for degradation.

    Science.gov (United States)

    Salah, Zaidoun; Cohen, Sherri; Itzhaki, Ella; Aqeilan, Rami I

    2013-12-15

    Proper regulation of cell proliferation, cell apoptosis, and cell death are vital for the development and survival of living organisms. Failure or dysfunction of any of these processes can have devastating effects, including cancer. The Hippo pathway, first discovered in Drosophila, has been found to be a major growth-regulatory signaling pathway that controls these crucial processes and has been implicated in cell-progress regulation and organ size determination. Abnormal regulation of this pathway has been found in several cancer types. However, the mechanisms that regulate the pathway and its core members yet have to be elucidated. One of the main core components of this pathway is LATS1, a serine/threonine kinase. Therefore, understanding how LATS1 activity is regulated is expected to shed light on new mechanisms that regulate the Hippo pathway. In the current work, we identified several potential LATS1 regulators and proved that NEDD4 E3 ubiquitin ligase controls LATS1 stability. We demonstrate that NEDD4 directly interacts with LATS1, leading to ubiquitination and decreased levels of LATS1 and, thus, increased YAP localization in the nucleus, which subsequently increases the transcriptional activity of YAP. As such, we show that NEDD4 acts as an additional regulator of the Hippo pathway on the protein level via interactions between WW domain-containing and PPxY motif-containing proteins. These findings might be applied in the development of new therapeutic approaches through the activation of LATS1.

  6. Arctigenin promotes degradation of inducible nitric oxide synthase through CHIP-associated proteasome pathway and suppresses its enzyme activity.

    Science.gov (United States)

    Yao, Xiangyang; Li, Guilan; Lü, Chaotian; Xu, Hui; Yin, Zhimin

    2012-10-01

    Arctigenin, a natural dibenzylbutyrolactone lignan compound, has been reported to possess anti-inflammatory properties. Previous works showed that arctigenin decreased lipopolysaccharide (LPS)-induced iNOS at transcription level. However, whether arctigenin could regulate iNOS at the post-translational level is still unclear. In the present study, we demonstrated that arctigenin promoted the degradation of iNOS which is expressed under LPS stimulation in murine macrophage-like RAW 264.7 cells. Such degradation of iNOS protein is due to CHIP-associated ubiquitination and proteasome-dependency. Furthermore, arctigenin decreased iNOS phosphorylation through inhibiting ERK and Src activation, subsequently suppressed iNOS enzyme activity. In conclusion, our research displays a new finding that arctigenin can promote the ubiqitination and degradation of iNOS after LPS stimulation. iNOS activity regulated by arctigenin is likely to involve a multitude of crosstalking mechanisms.

  7. Isolation and characterization of Fenitrothion-degrading bacteria from pestachio gardens in Kerman Provinance

    Directory of Open Access Journals (Sweden)

    Mehrnosh Ghafari

    2014-07-01

    Full Text Available   Introduction : Pesticides with complex structure have high persistence in ecosystem and biosphere. Pesticides have harmful effects on farmlands, human and natural resources.   Materials and methods: In this study for isolation of pesticide-degrading bacteria (Fenitrothion soil samples were collected from pistachio gardens in Kerman province. Collected soil samples were enriched in Bushnell Hass medium with this pesticide as only carbon and energy source. Isolated bacteria were identified by amplification of 16S rDNA gene by PCR and sequencing .   Results : In this study three Fenitrothion -degrading bacterial strains were isolated. These isolated bacteria were identified as: Pseudomonas fluorescens strain F1 ، Bacillus cereus strain F3 and pseudomonas aeruginosa strain F4 . The effects of pesticides concentration on each dominant bacterial strain were investigated. For Fenitrothion degrading bacterium (F4 strain growth continue until 100 ppm and then decreased. The result of Gas Chromatography (GC analysis confirmed the biodegradation ability of selected bacterial strains .   Discussion and conclusion : The results of this study demonstrated that there is a diversity of pesticide-degrading bacteria (Fenitrothion in soil ecosystem farmlands of Kerman province. It is seemed by application of these pesticide-degrading bacteria in farmlands and using bioremediation technique the ecosystem contamination of pesticide can be decreased.

  8. [Isolation and characterization of a halotolerant p-nitroaniline degrading strain S8].

    Science.gov (United States)

    Song, Cai-Xia; Deng, Xin-Ping; Li, Tian; Xiao, Wei

    2014-03-01

    A bacterial strain S8 was isolated from pesticide-contaminated sludge, which showed ability of utilizing p-nitroaniline as the sole source of carbon and nitrogen for growth. Based on the morphology, the physiological and biochemical characteristics, and the 16S rDNA sequence analysis, the strain was identified as Bacillus subtilis. Strain S8 showed high efficiency in p-nitroaniline degradation. 65.6% and 55.8% of p-nitroaniline (60 mg x L(-1) and 120 mg x L(-1)) were degraded by this strain within 72 hours under the optimal conditions of 31degrees C and pH 6.0. Besides, strain S8 degraded 49.5% p-nitroaniline in 7% NaCl and 27.4% p-nitroaniline in 10% NaCl (72 h), which showed that the strain S8 had a high salinity tolerance. When the LC-MS method was used for identification of the biodegradation products, six kinds of degradation products were found, two of which were identified as phenol and hydroquinone. To date, this is the first report on the degradation of p-nitroaniline by Bacillus subtilis. These results suggest that S8 could be a potential candidate for treating p-nitroaniline-contaminated saline wastewater.

  9. Screening, selection and characterization of phytic acid degrading lactic acid bacteria from chicken intestine.

    Science.gov (United States)

    Raghavendra, Ponnala; Halami, Prakash M

    2009-07-31

    This study was undertaken to screen and select potent phytate degrading lactic acid bacteria and to evaluate their additional characteristic features. Forty lactic acid bacterial strains were isolated from different sources and screened for their ability to degrade myo-inositol hexaphosphate or IP(6) by cobalt chloride staining (plate assay) method, using calcium or sodium salt of phytic acid as substrate. All the forty isolates were able to degrade calcium phytate. However, only two Pediococcus pentosaceus strains (CFR R38 and CFR R35) were found to degrade sodium phytate. These strains showed phytase activity of 213 and 89 U at 50 degrees C, respectively and poor acid phosphatase activity. These strains were further evaluated for additional characteristic features. At pH 2, P. pentosaceus strains CFR R38 and CFR R35 showed 50.7 and 48.5 percentage survivability after 2 h of incubation respectively and they could also withstand 0.3% ox-bile. These cultures exhibited 54.6 and 44.8% of hydrophobicity to xylene, antibacterial activity against food borne pathogens and possessed beta-galactosidase activity. The resistance pattern to several antibiotics was also analyzed. The present study indicates that these strains, having phytate degrading ability and other characteristic features can be exploited as starter cultures in fermented foods to improve the mineral bioavailability.

  10. Isolation and characterization of phenol degrading yeasts from wastewater in the coking plant of Zarand, Kerman.

    Science.gov (United States)

    Karimi, Maryam; Hassanshahian, Mehdi

    2016-01-01

    Phenol and phenolic compounds are environmental pollutants present in industrial wastewaters such as coal tar, oil refineries and petrochemical plants. Phenol removal from industrial effluents is extremely important for the protection of environment. Usually, phenol degradation is carried out by physicochemical methods that are costly and produce hazardous metabolites. Recently, phenol biodegradation has been considered. Yeasts are the most important phenol biodegraders. In this study, the phenol-degrading yeast from environmental samples (soil and wastewater) was isolated from the coking plant of Zarand, Kerman. Then total heterotrophic yeasts were counted. The soil samples had higher rates of yeast degrader, in comparison to wastewater samples. After three passages, four yeasts (K1, K2, K7 and K11) that had the highest growth rate were selected for further study. Also, these yeasts were able to remove phenol measured by Gibbs reagent. The effect of four different concentrations of phenol (50, 125, 200 and 275) mgL(-1) was measured and three degradation patterns in these yeasts were observed. The hydrophobicity and emulsification activity were measured in all eleven yeasts. Finally, strong yeasts in phenol degrading yeasts were identified by molecular method using amplification of 18S rRNA gene region. The sequencing results showed that these isolated yeasts belonged to Candida tropicalis strain K1, Pichia guilliermondii strain K2, Meyerozyma guilliermondii strain K7 and C. tropicalis strain K11.

  11. Isolation and characterization of phenol degrading yeasts from wastewater in the coking plant of Zarand, Kerman

    Directory of Open Access Journals (Sweden)

    Maryam Karimi

    2016-03-01

    Full Text Available Abstract Phenol and phenolic compounds are environmental pollutants present in industrial wastewaters such as coal tar, oil refineries and petrochemical plants. Phenol removal from industrial effluents is extremely important for the protection of environment. Usually, phenol degradation is carried out by physicochemical methods that are costly and produce hazardous metabolites. Recently, phenol biodegradation has been considered. Yeasts are the most important phenol biodegraders. In this study, the phenol-degrading yeast from environmental samples (soil and wastewater was isolated from the coking plant of Zarand, Kerman. Then total heterotrophic yeasts were counted. The soil samples had higher rates of yeast degrader, in comparison to wastewater samples. After three passages, four yeasts (K1, K2, K7 and K11 that had the highest growth rate were selected for further study. Also, these yeasts were able to remove phenol measured by Gibbs reagent. The effect of four different concentrations of phenol (50, 125, 200 and 275 mg L−1 was measured and three degradation patterns in these yeasts were observed. The hydrophobicity and emulsification activity were measured in all eleven yeasts. Finally, strong yeasts in phenol degrading yeasts were identified by molecular method using amplification of 18S rRNA gene region. The sequencing results showed that these isolated yeasts belonged to Candida tropicalis strain K1, Pichia guilliermondii strain K2, Meyerozyma guilliermondii strain K7 and C. tropicalis strain K11.

  12. Isolation and characterization of aniline degradation slightly halophilic bacterium, Erwinia sp. Strain HSA 6.

    Science.gov (United States)

    Li, Junmin; Jin, Zexin; Yu, Binbin

    2010-07-20

    The isolated strain HSA6 is classified as Erwinia amylovora based on 16S rDNA sequence and the morphological and physiological properties. Strain HSA6 is the first reported E. amylovora in pure culture growing with aniline as sole electron donor and carbon source. The suitable pH for strain HSA6 is wide (from 5 to 11). Strain HSA6 is slightly halophilic with growth occurring at 0-10% (v/v) NaCl, and the suitable NaCl concentration for strain HSA6 is from 0% to 6%. The number of bacteria appeared to decrease with an increase in aniline concentration. The number of bacteria appeared to be constant as the wastewater concentration increased from 0% to 20%. However, the number of cells decreased with an increase in wastewater concentration from 30% to 50% and grew very slowly at 50%. The degradation rate of aniline was 100% at 0.5% aniline concentration after 24 h culture. The degradation rate of aniline was found to descend as the concentration of aniline increased from 0.5% to 3% and rose as the culture time increased. Strain HSA6 contains a plasmid with molecular weight higher than 42 kDA. Plasmid curing test and quantitative degradation test showed that strain requires the plasmid for aniline degradation. The gene cluster degrading aniline was determined in the plasmid by PCR amplification.

  13. Magnetic α-Fe2O3/MCM-41 nanocomposites: preparation, characterization, and catalytic activity for methylene blue degradation.

    Science.gov (United States)

    Ursachi, Irina; Stancu, Alexandru; Vasile, Aurelia

    2012-07-01

    Catalysts based on nanosized magnetic iron oxide stabilized inside the pore system of ordered mesoporous silica MCM-41 have been prepared. The obtained materials were characterized by powder X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), vibrating sample magnetometer (VSM), and N(2) adsorption-desorption isotherm. XRD analysis showed that the obtained materials consist from the pure hematite crystalline phase (α-Fe(2)O(3)) dispersed within ordered mesoporous silica MCM-41. Magnetic measurements show that the obtained nanocomposites exhibit at room temperature weak ferromagnetic behavior with slender hysteresis. The catalytic activity of the magnetic α-Fe(2)O(3)/MCM-41 nanocomposites was evaluated by the degradation of methylene blue (MB) aqueous solution. For this purpose, an ultrasound-assisted Fenton-like process was used. The effect of solution pH on degradation of MB was investigated. The results indicated that US-H(2)O(2)-α-Fe(2)O(3)/MCM-41 nanocomposite system is effective for the degradation of MB, suggesting its great potential in removal of dyes from wastewater. It was found that the degradation rate of MB increases with decrease in the pH value of the solution.

  14. Functional and pharmacological characterization of the natriuretic peptide-dependent lipolytic pathway in human fat cells.

    Science.gov (United States)

    Moro, Cedric; Galitzky, Jean; Sengenes, Coralie; Crampes, François; Lafontan, Max; Berlan, Michel

    2004-03-01

    A lipolytic pathway involving natriuretic peptides has recently been discovered in human fat cells. Its functional characteristics and the interactions of the atrial natriuretic peptide (ANP)-induced effects with adrenergic and insulin pathways were studied. Characterization of the action of ANP antagonists, i.e., A71915, anantin, S-28-Y (Ser-28-Tyr, a synthesized peptide), and HS-142-1 (a microbial polysaccharide), was performed. Lipolytic assays and intracellular cGMP and cAMP determinations were performed on isolated fat cells. Cell membranes were used for binding studies. At low concentrations ANP and isoproterenol [beta-adrenergic receptor (beta-AR) agonist] exerted additive lipolytic effects. The alpha(2)-AR pathway did not interfere with that of ANP. Lipolytic effects of ANP were unaltered by a 2-h pretreatment of fat cells with insulin, whereas beta-AR-induced lipolysis was reduced. Homologous desensitization occurred for ANP-dependent lipolytic pathways. Dendroapsis natriuretic peptide exhibited a similar maximal effect but a 10-fold higher lipolytic potency than ANP and mini-ANP (the shortest form of ANP). The antagonist A71915 exhibited competitive antagonistic properties with a pA(2) value of 7.51. Anantin displayed noncompetitive antagonism and exerted an inhibitory action on basal and beta-adrenergic receptor-induced lipolytic response. S-28-Y exhibited antagonist potencies toward ANP-induced lipolysis and behaved as a partial lipolytic agonist with a lower pD(2) value (7.4 +/- 0.2) than ANP (9.4 +/- 0.3). HS-142-1 exerted the weakest antagonistic effects. The results demonstrate that ANP-dependent effects do not interfere with beta- and alpha(2)-adrenergic pathways in human fat cells. They are unaffected by insulin pretreatments of fat cells but undergo desensitization. In the search of potent and specific natriuretic peptide receptor-A antagonist, in the human fat cell, A71915 was the only reliable one found.

  15. Heterologous expression and characterization of bacterial 2-C-methyl-d-erythritol-4-phosphate pathway in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Carlsen, Simon; Ajikumar, Parayil Kumaran; Formenti, Luca Riccardo;

    2013-01-01

    the engineering of Escherichia coli genes encoding the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway into the genome of Saccharomyces cerevisiae and the characterization of intermediate metabolites synthesized by the MEP pathway in yeast. Our UPLC-MS analysis of the MEP pathway metabolites from engineered...... yeast showed that the pathway is active until the synthesis of 2-C-methyl-d-erythritol-2,4-cyclodiphosphate, but appears to lack functionality of the last two steps of the MEP pathway, catalyzed by the [4Fe–4S] iron sulfur cluster proteins encoded by ispG and ispH. In order to functionalize the last two...... steps of the MEP pathway, we co-expressed the genes for the E. coli iron sulfur cluster (ISC) assembly machinery. By deleting ERG13, thereby incapacitating the mevalonate pathway, in conjunction with labeling experiments with U–13C6 glucose and growth experiments, we found that the ISC assembly...

  16. Isolation and characterization of a bacterial strain that efficiently degrades sex steroid hormones

    Institute of Scientific and Technical Information of China (English)

    JI Shulan; LIU Zhipei; LIU Zhipeng; REN Haiyan

    2007-01-01

    A bacterial strain,ZY3,growing on sex steroid hormones as the sole source of carbon and energy was isolated from the sewage treatment plant of a prophylactic steroids factory.ZY3 degrades the 3-methoxy-17β-hyclroxy-1,3,5(10),8(9)-δ-4-estren (MHE).This strain was preliminarily identified as Raoultella sp.ZY3 according to its morphology and its 16S rRNA gene sequence.During the experimental period (72 h),the optimum temperature,pH and 3-MHE concentration for the degradation of hydride by the strain ZY3 were 35℃,10 and 10 mg/L,respectively.The degradation rate of the sex steroid hormones increased to 87% and 85% after the addition of maltose and peptone,respectively.

  17. Characterizing pesticide sorption and degradation in macro scale biopurification systems using column displacement experiments.

    Science.gov (United States)

    De Wilde, Tineke; Spanoghe, Pieter; Mertens, Jan; Sniegowksi, Kristel; Ryckeboer, Jaak; Jaeken, Peter; Springael, Dirk

    2009-04-01

    The efficiency of biopurification systems to treat pesticide-contaminated water was previously studied in microcosms. To validate the obtained results, macrocosm systems were set-up. Four pesticides (linuron, isoproturon, bentazone, and metalaxyl) were continuously applied to ten different organic substrate mixes. Retention of the pesticides was similar and in some cases slightly lower in the macrocosms compared to the microcosms. Differences in retention between the different mixes were however minimal. Moreover, the classification of the retention strength of the pesticides was identical to that observed in microcosms: linuron>isoproturon>metalaxyl>bentazone. Monod kinetics were used to describe delayed degradation, which occurred for isoproturon, metalaxyl and bentazone. No breakthrough of linuron was observed, thus, this pesticide was appointed as the most retained and/or degraded pesticide, followed by isoproturon, metalaxyl and bentazone. Finally, most of the matrix mixes efficient in degrading or retaining pesticides were mixes containing dried cow manure.

  18. Characterizing pesticide sorption and degradation in macro scale biopurification systems using column displacement experiments

    Energy Technology Data Exchange (ETDEWEB)

    Wilde, Tineke de [Laboratory of Crop Protection Chemistry, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium)], E-mail: Tineke.DeWilde@UGent.be; Spanoghe, Pieter [Laboratory of Crop Protection Chemistry, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent (Belgium); Mertens, Jan; Sniegowksi, Kristel; Ryckeboer, Jaak [Division of Soil and Water Management, Faculty of Bioscience Engineering, K.U. Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee (Belgium); Jaeken, Peter [PCF-Royal Research Station of Gorsem, De Brede Akker 13, 3800 Sint-Truiden (Belgium); Springael, Dirk [Division of Soil and Water Management, Faculty of Bioscience Engineering, K.U. Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee (Belgium)

    2009-04-15

    The efficiency of biopurification systems to treat pesticide-contaminated water was previously studied in microcosms. To validate the obtained results, macrocosm systems were set-up. Four pesticides (linuron, isoproturon, bentazone, and metalaxyl) were continuously applied to ten different organic substrate mixes. Retention of the pesticides was similar and in some cases slightly lower in the macrocosms compared to the microcosms. Differences in retention between the different mixes were however minimal. Moreover, the classification of the retention strength of the pesticides was identical to that observed in microcosms: linuron > isoproturon > metalaxyl > bentazone. Monod kinetics were used to describe delayed degradation, which occurred for isoproturon, metalaxyl and bentazone. No breakthrough of linuron was observed, thus, this pesticide was appointed as the most retained and/or degraded pesticide, followed by isoproturon, metalaxyl and bentazone. Finally, most of the matrix mixes efficient in degrading or retaining pesticides were mixes containing dried cow manure. - Transport of pesticides in macrocosm containing organic substrates.

  19. Isolation and characterization of bacteria degrading polychlorinated biphenyls from transformer oil.

    Science.gov (United States)

    Rojas-Avelizapa, N G; Rodríguez-Vázquez, R; Martínez-Cruz, J; Esparza-García, F; Montes de Oca-García, A; Ríos-Leal, E; Fernández-Villagómez, G

    1999-01-01

    Polychlorinated biphenyls from transformer oil were degraded in liquid culture under aerobic conditions using a mixed bacterial culture isolated from a transformer oil sample with a high content of polychlorinated biphenyls and other hydrocarbons. Four strains were identified, three of them corresponded to genus Bacillus, the other one to Erwinia. Bacteria in the transformer oil could remove as much as 65% of polychlorinated biphenyls (88% W/V in the transformer oil). Additional data showed that the two isolated strains of B. lentus were able to grow on transformer oil and degrade polychlorinated biphenyls by 80 and 83%. Our results provide evidence that microorganisms occurring in transformer oil have the potential to degrade polychlorinated biphenyls.

  20. Synthesized TiO{sub 2}/ZSM-5 composites used for the photocatalytic degradation of azo dye: Intermediates, reaction pathway, mechanism and bio-toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Kefu; Hu, Xin-Yan [College of the Environment and Ecology, Xiamen University, Xiamen (China); Chen, Bor-Yann; Hsueh, Chung-Chuan [Department of Chemical and Materials Engineering, National I-Lan University, I-Lan, Taiwan (China); Zhang, Qian [Department of Environmental Engineering, National Taiwan University, Taipei, Taiwan (China); Wang, Jiajie; Lin, Yu-Jung [College of the Environment and Ecology, Xiamen University, Xiamen (China); Chang, Chang-Tang, E-mail: ctchang73222@gmail.com [Department of Environmental Engineering, National I-Lan University, I-Lan, Taiwan (China)

    2016-10-15

    Highlights: • The major photo-catalytic degradation pathway of azo-dye was elaborated according to the identification of by-products from GC–MS and IC analysis. • Comparative assessment on characteristics of abiotic and biotic dye decolorization was analyzed. • EDTA (hole scavengers) and t-BuOH (radical scavengers) were used to determine the main active oxidative species in the system. • The toxicity effects of degradation intermediates of Reactive Black 5 (RB5) on the cellular respiratory activity were assessed. - Abstract: In this study, a one-step solid dispersion method was used to synthesize titanium dioxide (TiO{sub 2})/Zeolite Socony Mobil-5 (ZSM-5) composites with substantially reduced time and energy consumption. A degradation efficiency of more than 95% was achieved within 10 min using 50% PTZ (synthesized TiO{sub 2}/ZSM-5 composites with TiO{sub 2} contents of 50 wt% loaded on ZSM-5) at pH 7 and 25 °C. The possible degradation pathway of azo-dye Reactive Black 5 (RB5) was investigated using gas chromatography–mass spectrometry and ion chromatography (IC). The bonds between the N atoms and naphthalene groups are likely attacked first and cleaved by hydroxyl radicals, ultimately resulting in the decolorization and mineralization of the azo dye. A comparative assessment of the characteristics of abiotic and biotic dye decolorization was completed. In addition, the toxicity effects of the degradation intermediates of azo-dye RB5 on cellular respiratory activity were analyzed. The bio-toxicity results showed that the decay rate constants of CO{sub 2} production from the azo-dye RB5 samples at different degradation times increased initially and subsequently decreased, indicating that intermediates of higher toxicity could adhere to the catalyst surface and gradually destroyed by further photocatalytic oxidation. Additionally, EDTA (hole scavengers) and t-BuOH (radical scavengers) were used to detect the main active oxidative species in the system

  1. Characterization and Comparison of Conducting Polyaniline Synthesized by Three Different Pathways

    Institute of Scientific and Technical Information of China (English)

    DONG Chaofang; XIAO Kui; CHEN Ting; LI Huiyan; LI Xiaogang

    2011-01-01

    The conducting polyaniline (PANI) prepared by three chemical oxidative polymerization pathways including microemulsion,emulsion and aqueous solution methods were studied and compared.Their structures,morphologies and properties were characterized using FT-IR,XRD,TEM and TGA.PANI particles formed in aqueous solution have a smallest size and doping HC(l) enables to increase their conductivity.In contrast,PANI particles prepared by emulsion method have the highest thermal stability and conductivity and more ordered morphology.Rather different from these two methods,microemulsion approach allows forming nanocomposite PANI with tube-like nanostructure.

  2. Oxidative degradation of decabromodiphenyl ether (BDE 209) by potassium permanganate: reaction pathways, kinetics, and mechanisms assisted by density functional theory calculations.

    Science.gov (United States)

    Shi, Jiaqi; Qu, Ruijuan; Feng, Mingbao; Wang, Xinghao; Wang, Liansheng; Yang, Shaogui; Wang, Zunyao

    2015-04-07

    This study found that decabromodiphenyl ether (BDE 209) could be oxidized effectively by potassium permanganate (KMnO4) in sulfuric acid medium. A total of 15 intermediate oxidative products were detected. The reaction pathways were proposed, which primarily included cleavage of the ether bond to form pentabromophenol. Direct oxidation on the benzene ring also played an important role because hydroxylated polybrominated diphenyl ethers (PBDEs) were produced during the oxidation process. The degradation occurred dramatically in the first few minutes and fitted pseudo-first-order kinetics. Increasing the water content decelerated the reaction rate, whereas increasing the temperature facilitated the reaction. In addition, density functional theory (DFT) was employed to determine the frontier molecular orbital (FMO) and frontier electron density (FED) of BDE 209 and the oxidative products. The theoretical calculation results confirmed the proposed reaction pathways.

  3. Isolation and Characterization of Phenanthrene Degrading Bacteria from Diesel Fuel-Contaminated Antarctic Soils

    Directory of Open Access Journals (Sweden)

    Alejandro Gran-Scheuch

    2017-08-01

    Full Text Available Antarctica is an attractive target for human exploration and scientific investigation, however the negative effects of human activity on this continent are long lasting and can have serious consequences on the native ecosystem. Various areas of Antarctica have been contaminated with diesel fuel, which contains harmful compounds such as heavy metals and polycyclic aromatic hydrocarbons (PAH. Bioremediation of PAHs by the activity of microorganisms is an ecological, economical, and safe decontamination approach. Since the introduction of foreign organisms into the Antarctica is prohibited, it is key to discover native bacteria that can be used for diesel bioremediation. By following the degradation of the PAH phenanthrene, we isolated 53 PAH metabolizing bacteria from diesel contaminated Antarctic soil samples, with three of these isolates exhibiting a high phenanthrene degrading capacity. In particular, the Sphingobium xenophagum D43FB isolate showed the highest phenanthrene degradation ability, generating up to 95% degradation of initial phenanthrene. D43FB can also degrade phenanthrene in the presence of its usual co-pollutant, the heavy metal cadmium, and showed the ability to grow using diesel-fuel as a sole carbon source. Microtiter plate assays and SEM analysis revealed that S. xenophagum D43FB exhibits the ability to form biofilms and can directly adhere to phenanthrene crystals. Genome sequencing analysis also revealed the presence of several genes involved in PAH degradation and heavy metal resistance in the D43FB genome. Altogether, these results demonstrate that S. xenophagum D43FB shows promising potential for its application in the bioremediation of diesel fuel contaminated-Antarctic ecosystems.

  4. Characterization of insulin-degrading enzyme-mediated cleavage of Aβ in distinct aggregation states.

    Science.gov (United States)

    Hubin, Ellen; Cioffi, Federica; Rozenski, Jef; van Nuland, Nico A J; Broersen, Kerensa

    2016-06-01

    To enhance our understanding of the potential therapeutic utility of insulin-degrading enzyme (IDE) in Alzheimer's disease (AD), we studied in vitro IDE-mediated degradation of different amyloid-beta (Aβ) peptide aggregation states. Our findings show that IDE activity is driven by the dynamic equilibrium between Aβ monomers and higher ordered aggregates. We identify Met(35)-Val(36) as a novel IDE cleavage site in the Aβ sequence and show that Aβ fragments resulting from IDE cleavage form non-toxic amorphous aggregates. These findings need to be taken into account in therapeutic strategies designed to increase Aβ clearance in AD patients by modulating IDE activity.

  5. Production and characterization of esterase in Lantinus tigrinus for degradation of polystyrene.

    Science.gov (United States)

    Tahir, Lubna; Ishtiaq Ali, Muhammad; Zia, Muhammad; Atiq, Naima; Hasan, Fariha; Ahmed, Safia

    2013-01-01

    Polystyrene is considered stable to biological degradation. Lantinus tigrinus isolated from wood sample produced esterase in growth medium under normal conditions. However, acidic medium, 37 degrees C temperature, presence of tween 80; and urea and yeast extract in mineral salt medium enhance the production of esterase and specific activity. Purified esterase was active at broad pH range and 45 degrees C. FTIR analysis confirmed that esterase produced by Lantinus tigrinus effectively degraded polystyrene film and broke macromolecules down to non-toxic molecules. This study concludes that the presence of Lantinus tigrinus at dumping sites can be exploited for waste management containing high molecular weight synthetic polymers.

  6. Biochemical characterization and bioinformatic analysis of two large multi-domain enzymes from Microbacterium aurum B8.A involved in native starch degradation

    NARCIS (Netherlands)

    Valk, Vincent

    2017-01-01

    Microbacterium aurum B8.A is a unique bacterium with the ability to degrade starch granules through pore formation. In this study two enzymes (MaAmyA and MaAmyB) which are involved in granular starch degradation and were specific for the M. aurum B8.A strain, have been characterized in detail. Both

  7. Roles of reactive chlorine species in trimethoprim degradation in the UV/chlorine process: Kinetics and transformation pathways.

    Science.gov (United States)

    Wu, Zihao; Fang, Jingyun; Xiang, Yingying; Shang, Chii; Li, Xuchun; Meng, Fangang; Yang, Xin

    2016-11-01

    The UV/chlorine process, which forms several reactive species including hydroxyl radicals (HO) and reactive chlorine species (RCS) to degrade contaminants, is being considered to be an advanced oxidation process. This study investigated the kinetics and mechanism of the degradation of trimethoprim (TMP) by the UV/chlorine process. The degradation of TMP was much faster by UV/chlorine compared to UV/H2O2. The degradation followed pseudo first-order kinetics, and the rate constant (k') increased linearly as the chlorine dosage increased from 20 μM to 200 μM and decreased as pH rose from 6.1 to 8.8. k' was not affected by chloride and bicarbonate but decreased by 50% in the presence of 1-mg/L NOM. The contribution of RCS, including Cl, Cl2(-) and ClO, to the degradation removal rate was much higher than that of HO and increased from 67% to 87% with increasing pH from 6.1 to 8.8 under the experimental condition. The increasing contribution of RCS to the degradation with increasing pH was attributable to the increase in the ClO concentration. Kinetic modeling and radical scavenging tests verified that ClO mainly attacked the trimethoxybenzyl moiety of TMP. RCS reacted with TMP much faster than HOCl/OCl(-) to form chlorinated products (i.e., m/z 325) and chlorinated disinfection byproducts such as chloroform, chloral hydrate, dichloroacetonitrile and trichloronitromethane. The hydroxylation and demethylation of m/z 325 driven by HO generated m/z 327 and m/z 341. Meanwhile, reactions of m/z 325 with HO and RCS/HOCl/OCl(-) generated dichlorinated and hydroxylated products (i.e., m/z 377). All the chlorinated products could be further depleted to produce products with less degree of halogenation in the UV/chlorine process, compared to dark chlorination. The acute toxicity to Vibrio fischeri by UV/chlorine was lower than chlorination at the same removal rate of TMP. This study demonstrated the importance of RCS, in particular, ClO, in the degradation of micropollutants

  8. Carbon/nitrogen-doped TiO2: New synthesis route, characterization and application for phenol degradation

    Directory of Open Access Journals (Sweden)

    Aboubakr M. Abdullah

    2016-03-01

    Full Text Available Porous nanocrystalline carbon and nitrogen (CN-doped TiO2 photocatalyst was prepared using carbon tetrachloride and polyaniline as precursors. The obtained powders were characterized by X-ray diffraction (XRD, X-ray photoelectron spectroscopy (XPS, scanning electron microscopy (SEM, Raman spectroscopy, Fourier transform infrared (FT-IR spectroscopy, and gravimetric analysis. The purpose of this work was to explore the state and location of nitrogen and carbon atoms introduced inside the TiO2 lattice and to study the exploitation of the photocatalytic activity of the CN-doped TiO2 for application in phenol degradation under UV illumination. After 30 min from the illumination onset, 64% and 57% of the phenol were degraded when the CN-doped TiO2 and TiO2 catalysts were used respectively.

  9. Synthesis, Characterization and Application of Cobalt Incorporated Mesoporous KIT-6 for the Visible Light Assisted Degradation of Methylene Blue.

    Science.gov (United States)

    Sanjini, N S; Winston, B; Velmathi, S

    2015-09-01

    Highly ordered mesoporous three dimensional la3d cobalt doped KIT-6 (Co-KIT-6) material was synthesized by the simple impregnation method and characterized by various physicochemical techniques such as X-ray diffraction, N2 adsorption-desorption isotherm, diffuse reflectance spectroscopy, Fourier transform infra-red spectroscopy and scanning electron microscopy. Larger pore diameter tuned with a narrow size distribution was observed for prepared Co-KIT-6. A series of Co-KIT-6 with different weight percentages solutions were prepared and the photocatalytic efficacy of the samples for degradation of methylene blue was studied under UV and visible light using UV-visible spectrophotometer. It was found that the catalytic activity depends on the amount of cobalt incorporated into the material. The complete degradation of dye was observed within 50 min. Different amount of catalyst and reaction time was considered for the optimization studies.

  10. Distinct Dual C-Cl Isotope Fractionation Patterns during Anaerobic Biodegradation of 1,2-Dichloroethane: Potential To Characterize Microbial Degradation in the Field.

    Science.gov (United States)

    Palau, J; Yu, R; Hatijah Mortan, S; Shouakar-Stash, O; Rosell, M; Freedman, D L; Sbarbati, C; Fiorenza, S; Aravena, R; Marco-Urrea, E; Elsner, M; Soler, A; Hunkeler, D

    2017-03-07

    This study investigates, for the first time, dual C-Cl isotope fractionation during anaerobic biodegradation of 1,2-dichloroethane (1,2-DCA) via dihaloelimination by Dehalococcoides and Dehalogenimonas-containing enrichment cultures. Isotopic fractionation of 1,2-DCA (εbulk(C) and εbulk(Cl)) for Dehalococcoides (-33.0 ± 0.4‰ and -5.1 ± 0.1‰) and Dehalogenimonas-containing microcosms (-23 ± 2‰ and -12.0 ± 0.8‰) resulted in distinctly different dual element C-Cl isotope correlations (Λ = Δδ(13)C/Δδ(37)Cl ≈ εbulk(C)/εbulk(Cl)), 6.8 ± 0.2 and 1.89 ± 0.02, respectively. Determined isotope effects and detected products suggest that the difference on the obtained Λ values for biodihaloelimination could be associated with a different mode of concerted bond cleavage rather than two different reaction pathways (i.e., stepwise vs concerted). Λ values of 1,2-DCA were, for the first time, determined in two field sites under reducing conditions (2.1 ± 0.1 and 2.2 ± 2.9). They were similar to the one obtained for the Dehalogenimonas-containing microcosms (1.89 ± 0.02) and very different from those reported for aerobic degradation pathways in a previous laboratory study (7.6 ± 0.1 and 0.78 ± 0.03). Thus, this study illustrates the potential of a dual isotope analysis to differentiate between aerobic and anaerobic biodegradation pathways of 1,2-DCA in the field and suggests that this approach might also be used to characterize dihaloelimination of 1,2-DCA by different bacteria, which needs to be confirmed in future studies.

  11. Characterization of degradation process of sucrase-isomaltase in rat jejunum with monoclonal-antibody-based enzyme-linked immunosorbent assay

    National Research Council Canada - National Science Library

    Goda, T; Quaroni, A; Koldovský, O

    1988-01-01

    ...]. To characterize further the process of sucrase-isomaltase degradation in jejunum, we determined the amounts of immunoreactive sucrase-isomaltase in rat jejunum by using a monoclonal-antibody-based...

  12. Purification and Characterization of a Nonylphenol (NP)-degrading Enzyme from Bacillus cereus.Frankland

    Institute of Scientific and Technical Information of China (English)

    YANG Ge; ZHANG Ying; BAI Yanfen

    2011-01-01

    An extracellular NP-degrading enzyme secreted by Bacillus cereus.Frankland was purified to homogeneity by a combination of ammonium sulfate precipitation,Phenyl-Sepharose hydrophobic-interaction chromatography and DEAE anion-exchange chromatography.On SDS(sodium dodecyl sulfate)-polyacrylamide gel electrophoresis analysis,the purified enzyme showed a relative molecular mass of 58.3 kDa.The depolymerzation of subunits was accompanied with the loss of NP-degrading enzyme activity,and removing denaturing factors by dialysis could restore the dimer structure and enzymatic activity.The enzyme had an isoelectric point of 5.5 and an optimal temperature of 60℃,and was the most active at pH 6.0.The enzymatic activity was stable at pH 4-8 and inhibited by Cu2+.TenN-terminal amino acids were determined to be ASVNSIKIGY,demonstrating that the purified enzyme was a novel one.The hydrolysis pattern of the purified enzyme indicated that the NP-degrading enzyme was an endo NP-degrading enzyme.The extraordinary thermo-stability provided the enzyme with a good prospect to be used as a new tool for clean-production process for textile industry.

  13. Nondestructive Examination (NDE) Detection and Characterization of Degradation Precursors, Technical Progress Report for FY 2012

    Energy Technology Data Exchange (ETDEWEB)

    Ramuhalli, P.; Meyer, R.M.; Fricke, J.M.; Prowant, M.S.; Coble, J.B.; Griffin, J.W.; Pitman, S.G.; Dahl, M.E.; Kafentzis, T.A.; Roosendaal, T.J.

    2012-09-01

    The overall objective of this project was to investigate the effectiveness of nondestructive examination (NDE) technology in detecting material degradation precursors by initiating and growing cracks in selected materials and using NDE methods to measure crack precursors prior to the onset of cracking. Nuclear reactor components are subject to stresses over time that are not precisely known and that make the life expectancy of components difficult to determine. To prevent future issues with the operation of these plants because of unforeseen failure of components, NDE technology is needed that can be used to identify and quantify precursors to macroscopic degradation of materials. Some of the NDE methods being researched as possible solutions to the precursor detection problem are magnetic Barkhausen noise, nonlinear ultrasonics, acoustic emission, eddy current measurements, and guided wave technology. In FY12, the objective was to complete preliminary assessment of advanced NDE techniques for sensitivity to degradation precursors, using prototypical degradation mechanisms in laboratory-scale measurements. This present document reports on the deliverable that meets the following milestone: M3LW-12OR0402143 – Report detailing an initial demonstration on samples from the crack-initiation tests will be provided (demonstrating acceleration of the work).

  14. Degradation of structurally characterized proteins injected into HeLa cells. Basic measurements

    Energy Technology Data Exchange (ETDEWEB)

    Rogers, S.W.; Rechsteiner, M.

    1988-12-25

    Thirty-five proteins of known x-ray structure were labeled by chloramine-T radioiodination or by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using red cell-mediated microinjection. Degradation rates of the injected proteins were then determined over the next 50 h by measuring the release of soluble isotope to the culture medium. Control experiments demonstrated that the measured rates were not compromised by proteolysis within RBCs, the presence of unfused RBCs, or degradation of protein released from RBCs to the medium. Degradation of some injected proteins was faster during the first 12 h after fusion than at later times, apparently a response of HeLa cells to trypsinization. However, all proteins exhibited first-order degradation rates between 24 and 48 h post injection. Except for seven proteins, stabilities measured during this interval were unaffected by the labeling procedure. Reductive methylation was used to choose among the seven discordant values, and half-lives for the 35 proteins ranged from 16 h for lysozyme to 214 h for yeast alcohol dehydrogenase. Since half-lives for six of the injected proteins closely match values obtained by in vivo measurements, we consider our estimates of the metabolic stabilities of the injected proteins to be generally accurate. Therefore, the half-lives obtained by microinjection should prove useful in the search for relationships between protein structure and intracellular stability.

  15. Characterization of three plant biomass-degrading microbial consortia by metagenomics- and metasecretomics-based approaches

    NARCIS (Netherlands)

    Jiménez, Diego Javier; Brossi, Maria Julia de Lima; Schuckel, Julia; Kracun, Stjepan Kresimir; Willats, William George Tycho; van Elsas, Jan Dirk

    2016-01-01

    The selection of microbes by enrichment on plant biomass has been proposed as an efficient way to develop new strategies for lignocellulose saccharification. Here, we report an in-depth analysis of soil-derived microbial consortia that were trained to degrade once-used wheat straw (WS1-M),

  16. Characterization of radiolytically generated degradation products in the strip section of a TRUEX flowsheet

    Energy Technology Data Exchange (ETDEWEB)

    Dean R. Peterman; Lonnie G. Olson; Gary S. Groenewold; Rocklan G. McDowell; Richard D. Tillotson; Jack D. Law

    2013-08-01

    This report presents a summary of the work performed to meet the FCRD level 2 milestone M3FT-13IN0302053, “Identification of TRUEX Strip Degradation.” The INL radiolysis test loop has been used to identify radiolytically generated degradation products in the strip section of the TRUEX flowsheet. These data were used to evaluate impact of the formation of radiolytic degradation products in the strip section upon the efficacy of the TRUEX flowsheet for the recovery of trivalent actinides and lanthanides from acidic solution. The nominal composition of the TRUEX solvent used in this study is 0.2 M CMPO and 1.4 M TBP dissolved in n-dodecane and the nominal composition of the TRUEX strip solution is 1.5 M lactic acid and 0.050 M diethylenetriaminepentaacetic acid. Gamma irradiation of a mixture of TRUEX process solvent and stripping solution in the test loop does not adversely impact flowsheet performance as measured by stripping americium ratios. The observed increase in americium stripping distribution ratios with increasing absorbed dose indicates the radiolytic production of organic soluble degradation compounds.

  17. Characterization of hydrolytic degradation of polylactic acid/rice hulls composites in water at different temperatures

    Directory of Open Access Journals (Sweden)

    2011-02-01

    Full Text Available Hydrolytic degradations of polylactic acid/rice hulls (PLA/RH composites with various rice hulls contents due to water absorptions at 23, 51 and 69°C were investigated by studying the thermal properties, chemical composition, molecular weight, and morphology of the degraded products. The results have attested that the stability of PLA/RH composites in water depends slightly on rice hulls contents but it is significantly influenced by water temperature. Water absorption in 30 days at 23°C was between 0.87 and 9.25% depending on rice hull contents. However, at thermophilic temperatures, the water absorption and degradation of these products were increased significantly. Saturations were achieved in less than 25 and 9 days at 51°C and 69°C, respectively, while hydrolytic degradation was demonstrated by an increase in fragility and development of crystallinity. At 69°C, there were significant reductions of the decomposition and glass transition temperatures of the polymer by 13°C. These changes were associated with the reduction of the molecular weight of PLA from 153.1 kDa to ~10.7 kDa due to hydrolysis of its ester group.

  18. Barley aleurone cell death is not apoptotic: characterization of nuclease activities and DNA degradation.

    Science.gov (United States)

    Fath, A; Bethke, P C; Jones, R L

    1999-11-01

    Barley aleurone cells undergo programmed cell death (PCD) when exposed to gibberellic acid (GA), but incubation in abscisic acid (ABA) prevent PCD. We tested the hypothesis that PCD in aleurone cells occurs by apoptosis, and show that the hallmark of apoptosis, namely DNA cleavage into 180 bp fragments, plasma membrane blebbing, and the formation of apoptotic bodies do not occur when aleurone cells die. We show that endogenous barley aleurone nucleases and nucleases present in enzymes used for protoplast preparation degrade aleurone DNA and that DNA degradation by these nucleases is rapid and can result in the formation of 180 bp DNA ladders. Methods are described that prevent DNA degradation during isolation from aleurone layers or protoplasts. Barley aleurone cells contain three nucleases whose activities are regulated by GA and ABA. CA induction and ABA repression of nuclease activities correlate with PCD in aleurone cells. Cells incubated in ABA remain alive and do not degrade their DNA, but living aleurone cells treated with GA accumulate nucleases and hydrolyze their nuclear DNA. We propose that barley nucleases play a role in DNA cleavage during aleurone PCD.

  19. The Use of a Combination of alkB Primers to Better Characterize the Distribution of Alkane-Degrading Bacteria.

    Directory of Open Access Journals (Sweden)

    Diogo Jurelevicius

    Full Text Available The alkane monooxygenase AlkB, which is encoded by the alkB gene, is a key enzyme involved in bacterial alkane degradation. To study the alkB gene within bacterial communities, researchers need to be aware of the variations in alkB nucleotide sequences; a failure to consider the sequence variations results in the low representation of the diversity and richness of alkane-degrading bacteria. To minimize this shortcoming, the use of a combination of three alkB-targeting primers to enhance the detection of the alkB gene in previously isolated alkane-degrading bacteria was proposed. Using this approach, alkB-related PCR products were detected in 79% of the strains tested. Furthermore, the chosen set of primers was used to study alkB richness and diversity in different soils sampled in Carmópolis, Brazil and King George Island, Antarctica. The DNA extracted from the different soils was PCR amplified with each set of alkB-targeting primers, and clone libraries were constructed, sequenced and analyzed. A total of 255 alkB phylotypes were detected. Venn diagram analyses revealed that only low numbers of alkB phylotypes were shared among the different libraries derived from each primer pair. Therefore, the combination of three alkB-targeting primers enhanced the richness of alkB phylotypes detected in the different soils by 45% to 139%, when compared to the use of a single alkB-targeting primer. In addition, a dendrogram analysis and beta diversity comparison of the alkB composition showed that each of the sampling sites studied had a particular set of alkane-degrading bacteria. The use of a combination of alkB primers was an efficient strategy for enhancing the detection of the alkB gene in cultivable bacteria and for better characterizing the distribution of alkane-degrading bacteria in different soil environments.

  20. Characterization of protein degradation in serum-based lubricants during simulation wear testing of metal-on-metal hip prostheses.

    Science.gov (United States)

    Maskiewicz, Victoria K; Williams, Paul A; Prates, Sarah J; Bowsher, John G; Clarke, Ian C

    2010-08-01

    A size exclusion high performance liquid chromatography (SEC-HPLC) method has been developed which is capable of separation and quantitation of bovine serum albumin (BSA) and bovine serum globulin (BSG) components of serum-based lubricant (SBL) solutions. This allowed characterization of the stability profiles of these proteins when acting as lubricants during hip wear simulation, and identification of wear-specific mechanisms of degradation. Using cobalt-chromium metal-on-metal (MOM) hip joints, it was observed that BSA remained stable for up to 3 days (215K cycles) of wear testing after which the protein degraded in a fairly linear fashion. BSG on the other hand, began to degrade immediately and in a linear fashion with a rate constant of 5% per day. Loss of both proteins occurred via the formation of high molecular weight aggregates which precipitated out of solution. No fragmentation of the polypeptide backbone of either protein was observed. Data obtained suggest that protein degradation was not due to microbial contamination, denaturation at the air-water interface, or frictional heating of articulating joint surfaces in these studies. We conclude that the primary source of protein degradation during MOM simulation testing occurs via high shear rates experienced by SBL solutions at articulating surfaces, possibly coupled with metal-protein interactions occurring as new and reactive metal surfaces are generated during wear testing. The development of this analytical methodology will allow new studies to clarify the role of SBL solutions in wear simulation studies and the interactions and lubricating properties of serum proteins with prosthetic surfaces other than MOM.

  1. Synthesis and characterization of zinc oxide nanorods and its photocatalytic activities towards degradation of 2,4-D.

    Science.gov (United States)

    Meenakshi, G; Sivasamy, A

    2017-01-01

    Semiconductor zinc oxide nanorods (NRZnO) were prepared by sol-gel technique using zinc acetate as the precursor and ammonia as the precipitating agent. The prepared photocatalyst were characterized by X-ray diffraction (XRD), Fourier Transform Infrared (FTIR), UV-Visible diffuse reflectance spectroscopy (UV-Vis-DRS), X-ray photoelectron spectroscopy (XPS), Field Emission-Scanning Electron Microscopy (FE-SEM), Atomic Force Microscopy (AFM), High Resolution-Tunneling Microscope (HR-TEM), Brunauer, Emmett and Teller (BET) and electron paramagnetic resonance spectroscopy (EPR) analysis. Particle size of the prepared photocatalyst was established by XRD and FE-SEM analysis. The morphology and the formation of uniform NRZnO was controlled by the temperature, entire concentration of precursors, duration of aging process. High surface roughness and porosity confirmed by AFM analysis. Band gap energy of the synthesized photocatalyst (3.2eV) was determined by using diffuse reflectance spectroscopy. The in-situ production of OH radicals by the prepared photocatalyst was confirmed by electron paramagnetic resonance spectroscopy (EPR) spin trapping technique. The photocatalytic activity of prepared NRZnO was evaluated by photo degradation of 2, 4-dichlorophenoxyacetic acid (2, 4-D) under UV and visible light irradiations. Experimental parameter such as effect of pH, catalyst dosage, initial 2, 4- D concentrations and addition of different electrolytes on the degradation of 2, 4-D was also studied in detail. Neutral pH was found to be the optimum and catalyst dosage of 30mg/10ml resulted in higher percentage of degradation. The photo degraded samples were analyzed by chemical oxygen demand (COD) analysis, UV-Visible spectroscopy. Reusability of the prepared photocatalyst was tested upto three cycles without affecting its performance. The experimental shown the rate of degradation follows pseudo-first order rate kinetics with respect to 2, 4 D. Copyright © 2016 Elsevier Inc

  2. The Use of a Combination of alkB Primers to Better Characterize the Distribution of Alkane-Degrading Bacteria

    Science.gov (United States)

    Jurelevicius, Diogo; Alvarez, Vanessa Marques; Peixoto, Raquel; Rosado, Alexandre S.; Seldin, Lucy

    2013-01-01

    The alkane monooxygenase AlkB, which is encoded by the alkB gene, is a key enzyme involved in bacterial alkane degradation. To study the alkB gene within bacterial communities, researchers need to be aware of the variations in alkB nucleotide sequences; a failure to consider the sequence variations results in the low representation of the diversity and richness of alkane-degrading bacteria. To minimize this shortcoming, the use of a combination of three alkB-targeting primers to enhance the detection of the alkB gene in previously isolated alkane-degrading bacteria was proposed. Using this approach, alkB-related PCR products were detected in 79% of the strains tested. Furthermore, the chosen set of primers was used to study alkB richness and diversity in different soils sampled in Carmópolis, Brazil and King George Island, Antarctica. The DNA extracted from the different soils was PCR amplified with each set of alkB-targeting primers, and clone libraries were constructed, sequenced and analyzed. A total of 255 alkB phylotypes were detected. Venn diagram analyses revealed that only low numbers of alkB phylotypes were shared among the different libraries derived from each primer pair. Therefore, the combination of three alkB-targeting primers enhanced the richness of alkB phylotypes detected in the different soils by 45% to 139%, when compared to the use of a single alkB-targeting primer. In addition, a dendrogram analysis and beta diversity comparison of the alkB composition showed that each of the sampling sites studied had a particular set of alkane-degrading bacteria. The use of a combination of alkB primers was an efficient strategy for enhancing the detection of the alkB gene in cultivable bacteria and for better characterizing the distribution of alkane-degrading bacteria in different soil environments. PMID:23825163

  3. Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum.

    Science.gov (United States)

    Du, Lei; Ma, Li; Qi, Feifei; Zheng, Xianliang; Jiang, Chengying; Li, Ailei; Wan, Xiaobo; Liu, Shuang-Jiang; Li, Shengying

    2016-03-18

    4-Cresol is not only a significant synthetic intermediate for production of many aromatic chemicals, but also a priority environmental pollutant because of its toxicity to higher organisms. In our previous studies, a gene cluster implicated to be involved in 4-cresol catabolism, creCDEFGHIR, was identified in Corynebacterium glutamicum and partially characterized in vivo. In this work, we report on the discovery of a novel 4-cresol biodegradation pathway that employs phosphorylated intermediates. This unique pathway initiates with the phosphorylation of the hydroxyl group of 4-cresol, which is catalyzed by a novel 4-methylbenzyl phosphate synthase, CreHI. Next, a unique class I P450 system, CreJEF, specifically recognizes phosphorylated intermediates and successively oxidizes the aromatic methyl group into carboxylic acid functionality via alcohol and aldehyde intermediates. Moreover, CreD (phosphohydrolase), CreC (alcohol dehydrogenase), and CreG (aldehyde dehydrogenase) were also found to be required for efficient oxidative transformations in this pathway. Steady-state kinetic parameters (Km and kcat) for each catabolic step were determined, and these results suggest that kinetic controls serve a key role in directing the metabolic flux to the most energy effective route.

  4. Characterization of a strain capable of degrading a herbicide mixture of quinclorac and bensulfuronmethyl

    Institute of Scientific and Technical Information of China (English)

    LU Zhen-Mei; LI Zi-Mu; SANG Li-Ya; MIN Hang

    2008-01-01

    A bacterial strain,designated as LS,was isolated from a contaminated soil and was found to be capable of utilizing quinelorac,bensulfuromnethyl,and a mixture of the two as carbon and energy sources for growth.Strain LS was identified as Ochrobactrum sp.based on its physiological-biochemical properties,16S rDNA sequences,and phylogenetic analysis.The extent of degradation of quinclorac and bensulfuronmethyl at initial concentrations of 1.5 and 0.1 g L-1 was 90% and 67%,respectively,as measured by high performance liquid chromatography (HPLC).When a herbicide mixture of 0.34 g L-1 quinelorac and 0.02 g L-1 bensulfuronmethyl was applied as carbon sources,quinclorac and bensulfuronmethyl were degraded at 95.7% and 67.5%,respectively.It appears that quinclorac is utilized more easily in a mixture than in a single state.The optimal temperature for growth of strain LS was 37℃.Strain LS grew well at pH 6 to 9 and had the highest degradation level for quinclorac and bensulfuronmethyl at an initial pH of 7 and 8,respectively.Addition of 0.25 g L-1 yeast extract could promote the growth and extent of degradation of quinclorac and bensulfuronmethyl by strain LS.Strain LS also showed the capability to degrade other aromatic compounds such as catechol,propisochlor,4-chloro-2-methylphenoxyacetic acid sodium (MCPA-Na) and imazethapy.The isolate LS shows a huge potential to be used in bioremediation for treating complex herbicide residues.

  5. Isolation and characterization of Microbulbifer species 6532A degrading seaweed thalli to single cell detritus particles.

    Science.gov (United States)

    Wakabayashi, Masayuki; Sakatoku, Akihiro; Noda, Fumio; Noda, Minoru; Tanaka, Daisuke; Nakamura, Shogo

    2012-02-01

    To reduce the volume of seaweed wastes and extract polysaccharides, seaweed-degrading bacteria were isolated from drifting macroalgae harvested along the coast of Toyama Bay, Japan. Sixty-four bacterial isolates were capable of degrading "Wakame" (Undaria pinnatifida) thallus fragments into single cell detritus (SCD) particles. Amongst these, strain 6532A was the most active degrader of thallus fragments, and was capable of degrading thallus fragments to SCD particles within a day. Although the sequence similarity of the 16S rRNA gene of strain 6532A was 100% similar to that of Microbulbifer elongatus JAMB-A7, several distinct differences were observed between strains, including motility, morphology, and utilization of D: -arabinose and gelatin. Consequently, strain 6532A was classified as a new Microbulbifer strain, and was designated Microbulbifer sp. 6532A. Strain 6532A was capable of degrading both alginate and cellulose in the culture medium, zymogram analysis of which revealed the presence of multiple alginate lyases and cellulases. To the best of our knowledge, this is the first study to directly demonstrate the existence of these enzymes in Microbulbifer species. Shotgun cloning and sequencing of the alginate lyase gene in 6532A revealed a 1,074-bp open reading frame, which was designated algMsp. The reading frame encoded a PL family seven enzyme composed of 358 amino acids (38,181 Da). With a similarity of 74.2%, the deduced amino acid sequence was most similar to a Saccharophagus enzyme (alg 7C). These findings suggest that algMsp in strain 6532A is a novel alginate lyase gene.

  6. Isolation and Characterization of Methyl Parathion-degrading Bacteria Based on Microbial Sensor Construction

    Directory of Open Access Journals (Sweden)

    GENG Fang-fang

    2014-12-01

    Full Text Available Methyl parathion (MP, a kind of typical organophosphates pesticides (OPs, is widely used as agricultural insecticides. However, due to their neurotoxic effects on humans, the elimination of OPs has become increasingly important. Microbial sensors are consisted of biological components and transducers. Owing to their attractive advantages including low cost, easy of miniaturization and excellent selectivity, they have been widely used for environmental analysis. In this paper, four novel bacterial strains capable of utilizing methyl parathion as the sole carbon source were isolated from pesticide contaminated soils. These four isolates were identified based on morphological characteristics and 16S rRNA gene sequences analysis, and their capability of degrading methyl parathion were investigated by high performance liquid chromatography. The highest degrading efficiency strain was selected for further study of degrading mechanism. The results indicated that degradation rate of these four strains were all over 78% after incubation at 30 ℃, pH 7.0 for 7 d with the original concentration of methyl parathion 50 mg·L-1. The highest degradation rate was up to 100%. 16S rRNA gene sequences indicated that strain MP-6 was affiliated into the genus klebsiella. The LC-MS results indicated that methyl parathion was hydrolyzed to dimethyl thiophosphoric acid and p-nitrophenol by MP-6. A little of p-nitrophenol molecules could be further metabolized to 4-nitrocatechol and 1, 2, 4-benzenetrio. The results indicated that based on detecting the potential signal of intermediate product p-nitrophenol, the strain MP-6 could be used to construct microbial sensors for determination of organophosphorus pesticides in environment.

  7. Characterization of the myometrial transcriptome and biological pathways of spontaneous human labor at term

    Science.gov (United States)

    Mittal, Pooja; Romero, Roberto; Tarca, Adi L.; Gonzalez, Juan; Draghici, Sorin; Xu, Yi; Dong, Zhong; Nhan-Chang, Chia-Ling; Chaiworapongsa, Tinnakorn; Lye, Stephen; Kusanovic, Juan Pedro; Lipovich, Leonard; Mazaki-Tovi, Shali; Hassan, Sonia S.; Mesiano, Sam; Kim, Chong Jai

    2011-01-01

    Aims To characterize the transcriptome of human myometrium during spontaneous labor at term. Methods Myometrium was obtained from women with (n=19) and without labor (n=20). Illumina® HumanHT-12 microarrays were utilized. Moderated t-tests and False Discovery Rate adjustment of p-values were applied. qRT-PCR was performed for a select set of differentially expressed genes in a separate set of samples. ELISA and Western Blot were utilized to confirm differential protein production in a third sample set. Results 1) 471 genes were differentially expressed; 2) Gene Ontology analysis indicated enrichment of 103 biological processes and 18 molecular functions including: a) inflammatory response; b) cytokine activity; and c) chemokine activity; 3) systems biology pathway analysis using Signaling Pathway Impact Analysis indicated 6 significant pathways: a) cytokine-cytokine receptor interaction; b) Jak-Stat signaling; and c) complement and coagulation cascades; d) NOD-like receptor signaling pathway; e) Systemic Lupus Erythematosus; and f) Chemokine signaling pathway; 3) qRT-PCR confirmed over-expression of prostaglandin-endoperoxide synthase-2 (PTGS2/COX2), heparin binding EGF-like growth factor (HBEGF), chemokine C-C motif ligand 2 (CCL2/MCP1), leukocyte immunoglobulin-like receptor, subfamily A member 5 (LILRA5/LIR9), IL-8, IL-6, chemokine C-X-C motif ligand 6 (CXCL6/GCP2), nuclear factor of kappa light chain gene enhancer in B-cells inhibitor zeta (NFKBIZ), suppressor of cytokine signaling 3 (SOCS3) and decreased expression of FK506 binding-protein 5 (FKBP5) and aldehyde dehydrogenase (ALDH2) in labor; 4) IL-6, CXCL6, CCL2 and SOCS3 protein expression was significantly higher in the term labor group compared to the term not in labor group. Conclusions Myometrium of women in spontaneous labor at term is characterized by a stereotypic gene expression pattern consistent with over-expression of the inflammatory response and leukocyte chemotaxis. Differential gene

  8. The renaissance of bacillosamine and its derivatives: pathway characterization and implications in pathogenicity.

    Science.gov (United States)

    Morrison, Michael J; Imperiali, Barbara

    2014-02-01

    Prokaryote-specific sugars, including N,N'-diacetylbacillosamine (diNAcBac) and pseudaminic acid, have experienced a renaissance in the past decade because of their discovery in glycans related to microbial pathogenicity. DiNAcBac is found at the reducing end of oligosaccharides of N- and O-linked bacterial protein glycosylation pathways of Gram-negative pathogens, including Campylobacter jejuni and Neisseria gonorrhoeae. Further derivatization of diNAcBac results in the nonulosonic acid known as legionaminic acid, which was first characterized in the O-antigen of the lipopolysaccharide (LPS) in Legionella pneumophila. Pseudaminic acid, an isomer of legionaminic acid, is also important in pathogenic bacteria such as Helicobacter pylori because of its occurrence in O-linked glycosylation of flagellin proteins, which plays an important role in flagellar assembly and motility. Here, we present recent advances in the characterization of the biosynthetic pathways leading to these highly modified sugars and investigation of the roles that each plays in bacterial fitness and pathogenicity.

  9. Unique nonstructural proteins of Pneumonia Virus of Mice (PVM) promote degradation of interferon (IFN) pathway components and IFN-stimulated gene proteins.

    Science.gov (United States)

    Dhar, Jayeeta; Barik, Sailen

    2016-12-01

    Pneumonia Virus of Mice (PVM) is the only virus that shares the Pneumovirus genus of the Paramyxoviridae family with Respiratory Syncytial Virus (RSV). A deadly mouse pathogen, PVM has the potential to serve as a robust animal model of RSV infection, since human RSV does not fully replicate the human pathology in mice. Like RSV, PVM also encodes two nonstructural proteins that have been implicated to suppress the IFN pathway, but surprisingly, they exhibit no sequence similarity with their RSV equivalents. The molecular mechanism of PVM NS function, therefore, remains unknown. Here, we show that recombinant PVM NS proteins degrade the mouse counterparts of the IFN pathway components. Proteasomal degradation appears to be mediated by ubiquitination promoted by PVM NS proteins. Interestingly, NS proteins of PVM lowered the levels of several ISG (IFN-stimulated gene) proteins as well. These results provide a molecular foundation for the mechanisms by which PVM efficiently subverts the IFN response of the murine cell. They also reveal that in spite of their high sequence dissimilarity, the two pneumoviral NS proteins are functionally and mechanistically similar.

  10. Degradation pathways of aniline in aqueous solutions during electro-oxidation with BDD electrodes and UV/H2O2 treatment.

    Science.gov (United States)

    Benito, Aleix; Penadés, Aida; Lliberia, Josep Lluis; Gonzalez-Olmos, Rafael

    2017-01-01

    In this work, it has been studied the mineralization of aniline, a toxic substance of low biodegradability typically found in many industrial wastewaters, through electro-oxidation using boron doped diamond (BDD) electrodes and photo-oxidation (UV photolysis and UV/H2O2 treatments). It was observed that in electro-oxidation and UV/H2O2, it was feasible to reach aniline mineralizations higher than 85%. Two different degradation routes have been observed during the aniline oxidation in these two treatments. The first route was the mineralization pathway, in which aniline was oxidized to CO2, water and nitrate. The second route was the polyaniline pathway in which polyanilines of high molecular weight are formed. The intermediate compounds involved in both degradation routes are different depending on the treatment used. In the electro-oxidation, denitrification processes were also observed. From an economical point of view, electro-oxidation of aniline using BDD electrodes is more interesting than UV/H2O2 due it has an 87% lower operational cost. So, electro-oxidation using BDD electrodes seems to be a more suitable technique for the mineralization of wastewater containing aniline than UV or H2O2 based technologies.

  11. Degradation of 2,4-dihydroxibenzoic acid by vacuum UV process in aqueous solution: Kinetic, identification of intermediates and reaction pathway

    Energy Technology Data Exchange (ETDEWEB)

    Azrague, Kamal [Laboratoire IMRCP, CNRS UMR 5623, University of Toulouse, 118 route de Narbonne, 31062 Toulouse (France); Department for Water and Environment, SINTEF, Klaebuveien 153, Trondheim 7465 (Norway); Pradines, Vincent; Bonnefille, Eric [Laboratoire IMRCP, CNRS UMR 5623, University of Toulouse, 118 route de Narbonne, 31062 Toulouse (France); Laboratoire LCC, CNRS, 205 route de Narbonne, F31077 Toulouse Cedex 4 (France); Claparols, Catherine [Laboratoire LCC, CNRS, 205 route de Narbonne, F31077 Toulouse Cedex 4 (France); Universite de Toulouse, UPS, Service Commun de Spectrometrie de Masse, 118 route de Narbonne, F31062 Toulouse Cedex 9 (France); Maurette, Marie-Therese [Laboratoire IMRCP, CNRS UMR 5623, University of Toulouse, 118 route de Narbonne, 31062 Toulouse (France); Benoit-Marquie, Florence, E-mail: florence@chimie.ups-tlse.fr [Laboratoire IMRCP, CNRS UMR 5623, University of Toulouse, 118 route de Narbonne, 31062 Toulouse (France)

    2012-10-30

    Highlights: Black-Right-Pointing-Pointer Degradation of 2,4-dihydroxybenzoic acid (DHBA) by vacuum UV photolysis of water. Black-Right-Pointing-Pointer V-UV Xe-excimer lamps produced essentially hydroxyl radicals (HO Degree-Sign ). Black-Right-Pointing-Pointer Identification of all intermediates formed allowed us to propose a reaction pathway. Black-Right-Pointing-Pointer This reaction pathway showed that DHBA reacts differently with HO Degree-Sign and h+. Black-Right-Pointing-Pointer DHBA would be used as a probe to determine which of these entities were involved. - Abstract: 2,4-Dihydroxybenzoic acid (2,4-DHBA) is found frequently as a pollutant in natural waters and represents a threat to water quality because it is a precursor to the formation of quinones which are highly toxic. The degradation of 2,4-DHBA using the vacuum UV photolysis of water has been investigated. Irradiation was carried out in an annular photoreactor equipped with a Xe-excimer lamp situated in the centre and emitting at 172 nm. The degradation kinetic followed a pseudo first order and the reaction has been found to be very heterogeneous, especially at low concentration. Impacts of oxygen or temperature have also been investigated but no effect has been shown. LC-MS and HPLC-UV combined with other analytical techniques allowed the identification of the formation of trihydroxybenzoiec acids and trihydroxybenzenes which underwent a ring opening, conducting to the formation of aliphatic products named {alpha}, {beta}, {delta} and {gamma}. These products were in turn degraded successively into maleiec acid, malic and succinic acid, malonic acid, glyoxalic acid and oxalic acid before reaching the complete mineralization in about 180 min. The proposed reaction pathway has shown to be very different from the one observed for the TiO{sub 2} photocatalysis which involves only holes (h{sup +}) without any formation of aromatic intermediates. The different behaviours of 2,4-DHBA towards the h

  12. Structural characterization of the enzymes composing the arginine deiminase pathway in Mycoplasma penetrans.

    Science.gov (United States)

    Gallego, Pablo; Planell, Raquel; Benach, Jordi; Querol, Enrique; Perez-Pons, Josep A; Reverter, David

    2012-01-01

    The metabolism of arginine towards ATP synthesis has been considered a major source of energy for microorganisms such as Mycoplasma penetrans in anaerobic conditions. Additionally, this pathway has also been implicated in pathogenic and virulence mechanism of certain microorganisms, i.e. protection from acidic stress during infection. In this work we present the crystal structures of the three enzymes composing the gene cluster of the arginine deiminase pathway from M. penetrans: arginine deiminase (ADI), ornithine carbamoyltransferase (OTC) and carbamate kinase (CK). The arginine deiminase (ADI) structure has been refined to 2.3 Å resolution in its apo-form, displaying an "open" conformation of the active site of the enzyme in comparison to previous complex structures with substrate intermediates. The active site pocket of ADI is empty, with some of the catalytic and binding residues far from their active positions, suggesting major conformational changes upon substrate binding. Ornithine carbamoyltransferase (OTC) has been refined in two crystal forms at 2.5 Å and 2.6 Å resolution, respectively, both displaying an identical dodecameric structure with a 23-point symmetry. The dodecameric structure of OTC represents the highest level of organization in this protein family and in M.penetrans it is constituted by a novel interface between the four catalytic homotrimers. Carbamate kinase (CK) has been refined to 2.5 Å resolution and its structure is characterized by the presence of two ion sulfates in the active site, one in the carbamoyl phosphate binding site and the other in the β-phosphate ADP binding pocket of the enzyme. The CK structure also shows variations in some of the elements that regulate the catalytic activity of the enzyme. The relatively low number of metabolic pathways and the relevance in human pathogenesis of Mycoplasma penetrans places the arginine deiminase pathway enzymes as potential targets to design specific inhibitors against this human

  13. Structural characterization of the enzymes composing the arginine deiminase pathway