WorldWideScience

Sample records for b6-dependent enzymatic activities

  1. Localized cranial hyperostosis of meningiomas: a result of neoplastic enzymatic activity?

    DEFF Research Database (Denmark)

    Heick, A.; Mosdal, C.; Klinken, Leif

    1993-01-01

    Neuropathology, alkaline phosphatase, cranial hyperostosis, meningioma, ossifying enzymatic activity......Neuropathology, alkaline phosphatase, cranial hyperostosis, meningioma, ossifying enzymatic activity...

  2. Vitamin B6-Dependent Enzymes in the Human Malaria Parasite Plasmodium falciparum: A Druggable Target?

    OpenAIRE

    Thales Kronenberger; Jasmin Lindner; Meissner, Kamila A.; Zimbres, Flávia M.; Coronado, Monika A.; Sauer, Frank M.; Isolmar Schettert; Carsten Wrenger

    2014-01-01

    Malaria is a deadly infectious disease which affects millions of people each year in tropical regions. There is no effective vaccine available and the treatment is based on drugs which are currently facing an emergence of drug resistance and in this sense the search for new drug targets is indispensable. It is well established that vitamin biosynthetic pathways, such as the vitamin B6 de novo synthesis present in Plasmodium, are excellent drug targets. The active form of vitamin B6, pyridoxal...

  3. Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

  4. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

  5. Artificial cytoskeletal structures within enzymatically active bio-inorganic protocells.

    Science.gov (United States)

    Kumar, Ravinash Krishna; Li, Mei; Olof, Sam N; Patil, Avinash J; Mann, Stephen

    2013-02-11

    The fabrication of enzymatically active, semi-permeable bio-inorganic protocells capable of self-assembling a cytoskeletal-like interior and undergoing small-molecule dephosphorylation reactions is described. Reversible disassembly of an amino acid-derived supramolecular hydrogel within the internalized reaction space is used to tune the enzymatic activity of the nanoparticle-bounded inorganic compartments. PMID:23027575

  6. Production of lipase extrated from aqueous waste: enzymatic activity kinetics

    Directory of Open Access Journals (Sweden)

    Tatianne Ferreira de Oliveira

    2014-12-01

    Full Text Available Lipases are an important group of enzymes with various applications in the food, chemical and pharmaceutical industry, besides having great interest for the treatment of effluents with high lipid content. The objective of this study was to isolate, characterize and select lipolytic bacteria that produce lipase from aqueous waste effluents and to study the enzymatic activity kinetics of the extract obtained via submerged fermentation. The results obtained are promising, being possible to isolate and characterize 23 lipase-producing microorganisms, mostly gram-positive bacteria, but after the fermentation step in a liquid medium, gram negative bacteria showed the highest enzymatic activity (56.72 U.L-1 for STP 2A` bacterium and 81.99 U.L-1 for R2B. In the enzymatic activity kinetic study with the selected bacterium (R2B, among the six variables (temperature, pH, minimal mineral medium, soybean oil, glucose and sodium nitrate, temperature was the one that most positively influenced the enzymatic activity, and the best results were obtained at 40°C. It was concluded that the enzyme extract obtained from environmental waste may be used to treat the effluent and contribute to reduce environmental impacts.

  7. Controlling the enzymatic activity of a restriction enzyme by light

    OpenAIRE

    Schierling, Benno; Noël, Ann-Josée; Wende, Wolfgang; Hien, Le Thi; Volkov, Eugeny; Kubareva, Elena; Oretskaya, Tatiana; Kokkinidis, Michael; Römpp, Andreas; Spengler, Bernhard; Pingoud, Alfred

    2009-01-01

    For many applications it would be desirable to be able to control the activity of proteins by using an external signal. In the present study, we have explored the possibility of modulating the activity of a restriction enzyme with light. By cross-linking two suitably located cysteine residues with a bifunctional azobenzene derivative, which can adopt a cis- or trans-configuration when illuminated by UV or blue light, respectively, enzymatic activity can be controlled in a reversible manner. T...

  8. Lipid protrusions membrane softness, and enzymatic activity

    DEFF Research Database (Denmark)

    Jensen, Morten Østergaard; Høyrup, P.; Callisen, T.H.;

    2004-01-01

    protrusion modes and mechanical softness of phospholipid bilayers and on the other side the activity of enzymes acting on lipid bilayers composed of different unsaturated lipids. Specifically, our experiments show a correlation between the bilayer bending rigidity and the apparent Arrhenius activation energy...

  9. Silica–enzyme–ionic liquid composites for improved enzymatic activity

    OpenAIRE

    Katsuya Kato; Yuki Kawachi; Hitomi Nakamura

    2014-01-01

    Trypsin and pepsin enzyme-catalyzed precipitation of silica, synthesized by sol–gel chemistry in an ionic liquid, produces a composite material that demonstrates high enzymatic activity. This study investigates the structural properties of this silica–enzyme–ionic liquid composite material that allows for the retention of enzyme hydrolysis and condensation activity. The composite was prepared from a mixture of organo-functionalized triethoxysilane and tetraethoxysilane in an ionic liquid via ...

  10. ENZYMATIC ACTIVITY OF FRESH WATER ACTINOMYCETES

    OpenAIRE

    Gunda Madan Mohan; Singara Charya, M. A.

    2012-01-01

    Fresh water systems represent a largely untapped source for isolation of novel microorganisms. Gram-positive actinomycetes are of special interest, since they are known to produce chemically diverse compounds with wide range of biological activities. Twenty four actinomycetes with distinct characteristics were isolated from three freshwater systems of Karimnagar, Andhra Pradesh, India viz: Lower Manair Dam, Manakondur Pond and Kothapally Pond. These isolates were screened for their antagonist...

  11. ASPECTS CONCERNING THE ENZYMATIC ACTIVITY IN SEVERAL THERMOACTINOMYCETE STRAINS

    Directory of Open Access Journals (Sweden)

    Simona Dunca

    2003-08-01

    Full Text Available In the thermoactinomycete strains subjected to examination the values of their recorded enzymatic activities (i.e. α-amy lase, protease, exo-β-1,4 – glucanase, endo -β-1,4 – glucanase and β-glucosidase were lower in the stationary cultures as compared to the stirred ones. The strain Thermomonospora fusca BB255 was found to be highly cellulase- producing and at the same time able to synthesize α-amy lases and proteases.

  12. Enzymatic and inhibiting activity in boar epididymal fluid

    Czech Academy of Sciences Publication Activity Database

    Davidová, Nina; Ren, Š.; Liberda, J.; Jonáková, Věra; Maňásková-Postlerová, Pavla

    Praha: Biotechnologický ústav AVČR, v.v.i, 2014 - (Pěknicová, J.), s. 1-82 [XXth Symposium of Biology and Immunology of Reproduction with international participation. Třešť (CZ), 22.05.2014-24.05.2014] R&D Projects: GA ČR(CZ) P503/12/1834; GA ČR(CZ) P502/14/05547S; GA MŠk(CZ) CZ1.05/1.1.00/02.0109 Institutional research plan: CEZ:AV0Z50520701 Institutional support: RVO:86652036 Keywords : enzymatic activity * inhibiting activity * epididymal fluid * proteinase Subject RIV: CE - Biochemistry

  13. Enzymatic activity of lipase in post-metamorphic phase bullfrogs

    Directory of Open Access Journals (Sweden)

    Braga Luís Gustavo Tavares

    2006-01-01

    Full Text Available The knowledge of the digestive system of bullfrogs is an important step for the determination of their nutritional requirements throughout growth phases. With the objective of evaluating the enzymatic activity of lipase in the intestinal content of bullfrogs (Rana catesbeiana Shaw, 1802, 100 animals with median weight of 3.6 g were distributed in stalls under controlled temperature and photoperiod. The frogs, selected at the post-metamorphic phase, received commercial extruded diet ad libitum throughout the 87-day experiment. The collections of the intestinal content were performed by the desensitization of the frogs in ice and water at 0ºC and subsequent isolation of the small intestine. Determination of lipase activity was performed with a commercial enzymatic kit (Lipase-Bioclin, MG, Brazil, first measured in samples taken at day three (3.46 UI. During the initial phase the bullfrog possesses low lipase hydrolysis capacity was found, having a specific activity of 217 UI mg-1. In the subsequent period both lipase activity and specific lipase activity continuously increased. Lipase activity as a function of bullfrog weight fell after day twenty and reached 0.33 UI g-1, for frogs of medium weight (179 g. Feed for bullfrogs at the post-metamorphic phase weighing more than 10 g can have larger amounts of ingredients containg lipids, due to the increased digestive capacity of these frogs.

  14. The biosynthesis of GDP-L-colitose: C-3 deoxygenation is catalyzed by a unique coenzyme B6-dependent enzyme.

    Science.gov (United States)

    Beyer, Noelle; Alam, Jenefer; Hallis, Tina M; Guo, Zhihong; Liu, Hung-wen

    2003-05-14

    l-Colitose (1) is a 3,6-dideoxyhexose found in the O-antigen of gram-negative lipopoly-saccharides. While the biosynthesis of many deoxysugars have previously been investigated, l-colitose is distinct in that it originates from GDP-d-mannose. In contrast, other 3,6-dideoxyhexoses arise from CDP-d-glucose. Therefore, the enzymes involved in the l-colitose biosynthetic pathway must be specifically tailored to utilize such a modified substrate. The mode for deoxygenation at C-3 of colitose is of particular interest because this conversion in other naturally occurring 3,6-dideoxyhexoses requires a pair of enzymes, E1 and E3, acting in concert. Interestingly, no E3 equivalent was identified in the five open reading frames of the col biosynthetic gene cluster from Yersinia pseudotuberculosis IVA. However, the gene product of colD showed moderate similarity with the E1 gene (ddhC/ascC) of the ascarylose pathway (27% identity and 42% similarity). Because E1 is a pyridoxamine 5'-phosphate (PMP)-dependent enzyme, it was thought that ColD might also utilize PMP. Indeed, turnover was observed during incubation of ColD with substrate in the presence of excess PMP, but not with pyridoxal 5'-phosphate (PLP). However, the rate of product formation increased by more than 40-fold when l-glutamate was included in the PLP incubation. The formation of alpha-ketoglutarate as a byproduct under these conditions clearly indicated that ColD functions as a transaminase, recognizing both PMP and PLP. In this paper, we propose a novel biosynthetic route for colitose, including the unprecedented C-3 deoxygenation performed solely by ColD. The utilization of PMP in a dehydration reaction is rare, but the combined deoxygenation-transamination activity makes ColD a unique enzyme. PMID:12733868

  15. PARP1 Val762Ala polymorphism reduces enzymatic activity

    International Nuclear Information System (INIS)

    Poly(ADP-ribose) polymerase 1 (PARP1) modifies a variety of nuclear proteins by poly(ADP-ribosyl)ation, and plays diverse roles in molecular and cellular processes. A common PARP1 single nucleotide polymorphism (SNP) at codon 762, resulting in the substitution of alanine (Ala) for valine (Val) in the catalytic domain has been implicated in susceptibility to cancer. To characterize the functional effect of this polymorphism on PARP1, we performed in vitro enzymatic analysis on PARP1-Ala762 and PARP1-Val762. We found that PARP1-Ala762 displayed 57.2% of the activity of PARP1-Val762 for auto-poly(ADP-ribosyl)ation and 61.9% of the activity of PARP1-Val762 for trans-poly(ADP-ribosyl)ation of histone H1. The kinetic characterization revealed that the K m of PARP1-Ala762 was increased to a 1.2-fold of the K m of PARP1-Val762 for trans-poly(ADP-ribosyl)ation. Thus, the PARP1 Val762Ala polymorphism reduces the enzymatic activity of PARP1 by increasing K m. This finding suggests that different levels of poly(ADP-ribosyl)ation by PARP1 might aid in understanding Cancer risk of carriers of the PARP1 Val762Ala polymorphism

  16. Dramatic enhancement of enzymatic activity in organic solvents by lyoprotectants

    Energy Technology Data Exchange (ETDEWEB)

    Dabulis, K.; Klibanov, A.M. (Massachusetts Inst. of Tech., Cambridge (United States))

    1993-03-05

    When seven different hydrolytic enzymes (four proteases and three lipases) were lyophilized from aqueous solution containing a ligand, N-Ac-L-Phe-NH[sub 2], their catalytic activity in anhydrous solvents was far greater (one to two orders of magnitude) than that of the enzymes lyophilized without the ligand. This ligand-induced activation was expressed regardless of whether the substrate employed in organic solvents structurally resembled the ligand. Furthermore, nonligand lyoprotectants [sorbitol, other sugars, and poly(ethylene glycol)] also dramatically enhanced enzymatic activity in anhydrous solvents when present in enzyme aqueous solution prior to lyophilization. The effects of the ligand and of the lyoprotectants were nonadditive, suggesting the same mechanism of action. Excipient-activated and nonactivated enzymes exhibited identical activities in water. Also, addition of the excipients directly to suspensions of nonactivated enzymes in organic solvents had no appreciable effect on catalytic activity. These observations indicate that the mechanism of the excipient-induced activation is based on the ability of the excipients to alleviate reversible denaturation of enzymes upon lyophilization. Activity enhancement induced by the excipients is displayed even after their removal by washing enzymes with anhydrous solvents. Subtilisin Carlsberg, lyophilized with sorbitol, was found to be a much more efficient practical catalyst than its regular' counterpart.

  17. ALTERED ENZYMATIC ACTIVITY OF LYSOZYMES BOUND TO VARIOUSLY SULFATED CHITOSANS

    Institute of Scientific and Technical Information of China (English)

    Hong-wei Wang; Lin Yuan; Tie-liang Zhao; He Huang; Hong Chen; Di Wu

    2012-01-01

    The purpose of this research is to investigate the effects of the variously sulfated chitosans on lysozyme activity and structure.It was shown that the specific enzymatic activity of lysozyme remained almost similar to the native protein after being bound to 6-O-sulfated chitosan (6S-chitosan) and 3,6-O-sulfated chitosan (3,6S-chitosan),but decreased greatly after being bound to 2-N-6-O-sulfated chitosan (2,6S-chitosan).Meanwhile,among these sulfated chitosans,2,6S-chitosan induced the greatest conformational change in lysozyme as indicated by the fluorescence spectra.These findings demonstrated that when sulfated chitosans of different structures bind to lysozyme,lysozyme undergoes conformational change of different magnitudes,which results in corresponding levels of lysozyme activity.Further study on the interaction of sulfated chitosans with lysozyme by surface plasmon resonance (SPR) suggested that their affinities might be determined by their molecular structures.

  18. Plasmin enzymatic activity in the presence of actin

    Directory of Open Access Journals (Sweden)

    Yusova E. I.

    2015-10-01

    Full Text Available Aim. To study the changes in the plasmin activity towards substrates with high and low molecular mass in the presence of actin. Methods. The proteins used for this investigation were obtained by affinity chromatography and gel-filtration. The plasmin enzymatic activity was determined by a turbidimetric assay and a chromogenic substrate-based assay. The enzyme linked immunosorbent assay and biotin-avidin-phosphatase system were used to study the interaction of plasminogen and its fragments with actin. Results. It was shown that G-actin causes 1.5-fold decrease in the rate of polymeric fibrin hydrolysis by plasmin and Glu-plasminogen activated by the tissue plasminogen activator. However, actin did not impede plasmin autolysis and had no influence on its amidase activity. We have studied an interaction of biotinylated Glu-plasminogen and its fragments (kringle 1-3, kringle 4 and mini-plasminogen with immobilized G-actin. Glu-plasminogen and kringle 4 had a high affinity towards actin (C50 is 113 and 117 nM correspondingly. Mini-plasminogen and kringe 4 did not bind to actin. A similar affinity of Glu-plasminogen and kringle 1-3 towards actin proves the involvement of the kringle 1-3 lysine-binding sites of the native plasminogen form in the actin interaction. Conclusions. Actin can modulate plasmin specificity towards high molecular mass substrates through its interaction with lysine-binding sites of the enzyme kringle domains. Actin inhibition of the fibrinolytic activity of plasmin is due to its competition with fibrin for thelysine binding sites of plasminogen/plasmin.

  19. Effect of Gypsum Application on Enzymatic Browning Activity in Lettuce

    Directory of Open Access Journals (Sweden)

    Prasit Chutichudet

    2009-01-01

    Full Text Available A comprehensive study to evaluate calcium, in terms of gypsum (CaSO4.2H2O by soil dressing application, on enzymatic browning activity of Polyphenol oxidase (PPO and internal qualities was tested on lettuce var. Grand Rapids under field conditions. A factorial in completely randomized design was arranged with four replications. The results showed that plants-treated with 50 mg kg-1 gypsum applied at 40 DAP had the maximal fresh weight of 25.83 g plant-1. The internal qualities of the lettuce at harvest showed that plants treated with 50 mg kg-1 gypsum had the maximal chlorophyll content (26.80 mg m-2, while all gypsum concentrations applied in this study, had less content of ascorbic acid than the control plants. Plants-treated with 100 mg kg-1 gypsum affected to the lowest level of PPO activity at week 3 after transplanting. Furthermore, gypsum application had no effect to biomass, leaf colour, the contents of phenolic and quinone in lettuce at harvesting stage.

  20. Testing the applicability of rapid on-site enzymatic activity detection for surface water monitoring

    Science.gov (United States)

    Stadler, Philipp; Vogl, Wolfgang; Juri, Koschelnik; Markus, Epp; Maximilian, Lackner; Markus, Oismüller; Monika, Kumpan; Peter, Strauss; Regina, Sommer; Gabriela, Ryzinska-Paier; Farnleitner Andreas, H.; Matthias, Zessner

    2015-04-01

    On-site detection of enzymatic activities has been suggested as a rapid surrogate for microbiological pollution monitoring of water resources (e.g. using glucuronidases, galactosidases, esterases). Due to the possible short measuring intervals enzymatic methods have high potential as near-real time water quality monitoring tools. This presentation describes results from a long termed field test. For twelve months, two ColiMinder devices (Vienna Water Monitoring, Austria) for on-site determination of enzymatic activity were tested for stream water monitoring at the experimental catchment HOAL (Hydrological Open Air Laboratory, Center for Water Resource Systems, Vienna University of Technology). The devices were overall able to follow and reflect the diverse hydrological and microbiological conditions of the monitored stream during the test period. Continuous data in high temporal resolution captured the course of enzymatic activity in stream water during diverse rainfall events. The method also proofed sensitive enough to determine diurnal fluctuations of enzymatic activity in stream water during dry periods. The method was able to capture a seasonal trend of enzymatic activity in stream water that matches the results gained from Colilert18 analysis for E. coli and coliform bacteria of monthly grab samples. Furthermore the comparison of ColiMinder data with measurements gained at the same test site with devices using the same method but having different construction design (BACTcontrol, microLAN) showed consistent measuring results. Comparative analysis showed significant differences between measured enzymatic activity (modified fishman units and pmol/min/100ml) and cultivation based analyses (most probable number, colony forming unit). Methods of enzymatic activity measures are capable to detect ideally the enzymatic activity caused by all active target bacteria members, including VBNC (viable but nonculturable) while cultivation based methods cannot detect VBNC

  1. First results on enzymatic activities in two salt marsh soils under different hydromorphic level and vegetation

    Directory of Open Access Journals (Sweden)

    Carmen Trasar-Cepeda

    2015-12-01

    Full Text Available Salt-marsh soils are soils characterized by non-permanent hydric saturation that, depending on factors like duration of submersion periods, are dominated by different salt-tolerant plant species. The composition of microbial communities is an essential component in trophic dynamics and biogeochemical processes in salt marshes, and determines the level of enzymatic activities, which catalyze the conversion of complex molecules into simpler ones. Despite of this, the enzymatic activities in marsh-soils has not yet been investigated. The aim of this study was to analyze the enzymatic activities in two soil profiles of marsh-soils under different water saturation level and dominated by different plant species [Juncus maritimus Lam and Spartina maritima (Curtis Fernald (Sp]. In both soils, the enzymatic activities were much lower than the levels typically found in terrestrial ecosystems. The enzymatic activities were measured both in air-dried and in re-moistened and incubated soil samples. In air-dried samples, the enzymatic activities were higher in Juncus than in Spartina soil and tended to decrease with depth, being sharper the decrease in Juncus than in Spartina soil. Re-moistened and pre-incubated soils showed a general increase in all the enzymatic activities and throughout the whole soil profile, especially in Spartina soils. Hydrolase activities showed a strong and positive relationship with organic matter content both in air-dried and in re-moistened soil samples, higher in these latter. In general, oxidoreductase activities only showed this relationship in re-moistened soil samples. More studies, preferably using freshly collected soil samples, are needed to understand the relationship between enzymatic activities and these environmental conditions.

  2. A novel Access to Nucleotide-activated Oligosaccharides by Enzymatic Synthesis

    Czech Academy of Sciences Publication Activity Database

    Nieder, V.; Gallego, R.; Kutzer, M.; Kamerling, J.; Vliegenthart, J.; Marx, S.; Křen, Vladimír; Elling, L.

    Hamburg, 2000. s. 152. [International Carbohydrate Symposium /20./. 27.08.2000-01.09.2000, Hamburg] Institutional research plan: CEZ:AV0Z5020903 Keywords : nucleotide * activated * enzymatic Subject RIV: EE - Microbiology, Virology

  3. First results on enzymatic activities in two salt marsh soils under different hydromorphic level and vegetation

    OpenAIRE

    Carmen Trasar-Cepeda; Diana Bello; Chiara Ferronato; Livia Vittori Antisari

    2015-01-01

    Salt-marsh soils are soils characterized by non-permanent hydric saturation that, depending on factors like duration of submersion periods, are dominated by different salt-tolerant plant species. The composition of microbial communities is an essential component in trophic dynamics and biogeochemical processes in salt marshes, and determines the level of enzymatic activities, which catalyze the conversion of complex molecules into simpler ones. Despite of this, the enzymatic activities in mar...

  4. Molecular dynamics study of enhanced Man5B enzymatic activity

    OpenAIRE

    Bernardi, Rafael C; Cann, Isaac; Schulten, Klaus

    2014-01-01

    Background Biofuels are a well-known alternative to the largely used fossil-derived fuels, however the competition with food production is an ethical dilemma. Fortunately a solution is offered by second-generation biofuels which can be produced from agricultural waste or, more specifically, from plant cell wall polysaccharides. The conversion process involves typically enzymatic hydrolysis of lignocellulosic biomass and then separation of its constituent sugars that are further fermented to p...

  5. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M.; Negro, M. J.; Saez, R.; Martin, C.

    1986-07-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  6. Enzymatic Synthesis and Anti-Allergic Activities of Curcumin Oligosaccharides

    OpenAIRE

    Hiroki Hamada; Kei Shimoda

    2010-01-01

    Curcumin 4'-O-glucooligosaccharides were synthesized by a two step-enzymatic method using almond β-glucosidase and cyclodextrin glucanotransferase (CGTase). Curcumin was glucosylated to curcumin 4'-O-β-D-glucopyranoside by almond β-glucosidase in 19% yield. Curcumin 4'-O-β-D-glucopyranoside was converted into curcumin 4'-O-β-glucooligosaccharides, i.e. 4'-O-β-maltoside (51%) and 4'-O-β-maltotrioside (25%), by further CGTase-catalyzed glycosylation. Curcumin 4'-O-β-glycosides showed suppressiv...

  7. Enzymatic Activity in the Gastrointestinal Tract of Pimelodus maculatus (Teleostei, Siluriformes) in Two Neotropical Reservoirs with Different Trophic Conditions

    OpenAIRE

    Silvana Duarte; Marcelo Bemquerer; Francisco Gerson Araújo

    2015-01-01

    Enzymatic activities for digestion of proteins and carbohydrates were compared among three organs of the digestive system of Pimelodus maculatus in two reservoirs with different trophic conditions during the winter of 2006. The aim was to test the hypothesis that enzymatic activity for the digestion of proteins and carbohydrates differed among organs and that such activities differ between the trophic state of the environment. Enzymatic activities were determined through the assays of specifi...

  8. Enzymatic activity measured by microcalorimetry in soil amended with organic residues

    Directory of Open Access Journals (Sweden)

    Karina Cenciani

    2011-08-01

    Full Text Available Enzymatic activity is an important property for soil quality evaluation. Two sequences of experiments were carried out in order to evaluate the enzymatic activity in a soil (Rhodic Eutrudox amended with cattle manure, earthworm casts, or sewage sludges from the municipalities of Barueri and Franca. The activity of commercial enzymes was measured by microcalorimetry in the same soil samples after sterilization. In the first experiment, the enzyme activities of cellulase, protease, and urease were determined in the soil samples during a three month period. In the second sequence of experiments, the thermal effect of the commercial enzymes cellulase, protease, and urease on sterilized soil samples under the same tretaments was monitored for a period of 46 days. The experimental design was randomized and arranged as factorial scheme in five treatments x seven samplings with five replications. The treatment effects were statistically evaluated by one-way analysis of variance. Tukey´s test was used to compare means at p < 0.05. The presence of different sources of organic residues increased the enzymatic activity in the sampling period. Cattle manure induced the highest enzymatic activity, followed by municipal sewage sludge, whereas earthworm casts induced the lowest activity, but differed from control treatment. The thermal effect on the enzyme activity of commercial cellulase, protease, and urease showed a variety of time peaks. These values probably oscillated due to soil physical-chemical factors affecting the enzyme activity on the residues.

  9. Differential enzymatic activity of common haplotypic versions of the human acidic mammalian chitinase protein

    NARCIS (Netherlands)

    M.A. Seibold; T.A. Reese; S. Choudhry; M.T. Salam; K. Beckman; C. Eng; A. Atakilit; K. Meade; M. Lenoir; H.G. Watson; S. Thyne; R. Kumar; K.B. Weiss; L.C. Grammer; P. Avila; R.P. Schleimer; J.V. Fahy; J. Rodriguez-Santana; W. Rodriguez-Cintron; R.G. Boot; D. Sheppard; F.D. Gilliland; R.M. Locksley; E.G. Burchard

    2009-01-01

    Mouse models have shown the importance of acidic mammalian chitinase activity in settings of Th2 inflammation. AMCase enzymatic activity was necessary for most of the inflammation present in an asthma mouse model. However, in settings of chitin exposure, AMCase-mediated cleavage of the inflammatory

  10. Thrombolytic efficacy and enzymatic activity of rt-PA-loaded echogenic liposomes.

    Science.gov (United States)

    Bader, Kenneth B; Bouchoux, Guillaume; Peng, Tao; Klegerman, Melvin E; McPherson, David D; Holland, Christy K

    2015-08-01

    Echogenic liposomes (ELIP), that can encapsulate both recombinant tissue-type plasminogen activator (rt-PA) and microbubbles, are under development to improve the treatment of thrombo-occlusive disease. However, the enzymatic activity, thrombolytic efficacy, and stable cavitation activity generated by this agent has yet to be evaluated and compared to another established ultrasound-enhanced thrombolytic scheme. A spectrophotometric method was used to compare the enzymatic activity of the rt-PA incorporated into ELIP (t-ELIP) to that of rt-PA. An in vitro flow model was employed to measure the thrombolytic efficacy and dose of ultraharmonic emissions from stable cavitation for 120-kHz ultrasound exposure of three treatment schemes: rt-PA, rt-PA and the perfluorocarbon-filled microbubble Definity(®), and t-ELIP. The enzymatic activity of rt-PA incorporated into t-ELIP was 28 % that of rt-PA. Thrombolytic efficacy of t-ELIP or rt-PA and Definity(®) was equivalent when the dose of t-ELIP was adjusted to produce comparable enzymatic activity. Sustained bubble activity was nucleated from Definity but not from t-ELIP exposed to 120-kHz ultrasound. These results emphasize the advantages of encapsulating a thrombolytic and the importance of incorporating an insoluble gas required to promote sustained, stable cavitation activity. PMID:25829338

  11. PARTIAL CHARACTERIZATION OF ENZYMATIC ACTIVITIES PRODUCED BY A WILD STRAIN OF A. NIGER

    Directory of Open Access Journals (Sweden)

    María Martos

    2012-12-01

    Full Text Available Aspergillus niger, isolated from decay citrus peels in the province of Misiones, was able to produce pectinases by submerged fermentation. The enzymatic extract exhibited polygalacturonase, pectinesterase and lyase activities. Others enzymes capable of degrading cell wall polymers were also detected in the enzymatic extract such as cellulases and xylanases. Polygalacturonase was an endo-polygalacturonase. The enzyme exhibited a maximal activity at pH range between 4.5 to 5.0, was stable in the pH range from 2.5 to 5.5 and remained unchanged when was incubated at temperatures lower than 50 ºC. The fungi produced three PG isoenzymes. The enzymatic extract was able to clarify apple juice. The results observed make the pectinolytic enzymes produced by A. niger appropriate for future application in fruit juice processing industries.

  12. Plant oligoadenylates: enzymatic synthesis, isolation, and biological activities

    International Nuclear Information System (INIS)

    An enzyme that converts [3H, 32P]ATP, with a 3H:32P ratio of 1:1, to oligoadenylates with the same 3H:32P ratio was increased in plants following treatment with human leukocyte interferon or plant antiviral factor or inoculation with tobacco mosaic virus. The enzyme was extracted from tobacco leaves, callus tissue cultures, or cell suspension cultures. The enzyme, a putative plant oligoadenylate synthetase, was immobilized on poly(rI) . poly(rC)-agarose columns and converted ATP into plant oligoadenylates. These oligoadenylates were displaced from DEAE-cellulose columns with 350 mM KCl buffer, dialyzed, and further purified by high-performance liquid chromatography (HPLC) and DEAE-cellulose gradient chromatography. In all steps of purification, the ratio of 3H:32P in the oligoadenylates remained 1:1. The plant oligoadenylates isolated by displacement with 350 mM KCl had a molecular weight greater than 1000. The plant oligoadenylates had charges of 5- and 6-. HPLC resolved five peaks, three of which inhibited protein synthesis in reticulocyte and wheat germ systems. Partial structural elucidation of the plant oligoadenylates has been determined by enzymatic and chemical treatments. An adenylate with a 3',5'-phosphodiester and/or a pyrophosphoryl linkage with either 3'- or 5'-terminal phosphates is postulated on the basis of treatment of the oligoadenylates with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase and acid and alkaline hydrolyses. The plant oligoadenylates at 8 X 10(-7) M inhibit protein synthesis by 75% in lysates from rabbit reticulocytes and 45% in wheat germ cell-free systems

  13. Enzymatic hydrolysis of rice protein with papain and antioxidation activity of hydrolysate

    Science.gov (United States)

    The enzymatic hydrolysis technology of rice protein and the antioxidant activity of the hydrolysate were studied. Substrate concentration,enzyme dose,pH value and temperature were selected as factors to optimize the hydrolysis parameters with single—factor and orthogonal tests. Results show the opti...

  14. A comparison of protein kinases inhibitor screening methods using both enzymatic activity and binding affinity determination

    DEFF Research Database (Denmark)

    Rudolf, Amalie Frederikke; Skovgaard, Tine; Knapp, Stefan; Jensen, Lars Juhl; Berthelsen, Jens

    2014-01-01

    Binding assays are increasingly used as a screening method for protein kinase inhibitors; however, as yet only a weak correlation with enzymatic activity-based assays has been demonstrated. We show that the correlation between the two types of assays can be improved using more precise screening...

  15. The variations of enzymatic activity of pepsin preparation by γ-irradiation

    International Nuclear Information System (INIS)

    Effect of γ-irradiation on the enzymatic activity of raw pepsin and some saccharated pepsin preparations were studied in the dose range from 0 to 300 kGy. As a result, the apparent reduction rate of saccharated pepsin preparations is less than of raw pepsin. K values of raw and saccharated pepsins were 0.014 and 0.0040-0.0061, and G values of raw and saccharated pepsins were 3.98 and 1.13-1.73, respectively. The lower K and G values of saccharated pepsin than those of raw pepsin seem to be due to radiolytic products of lactose in the preparations as an excipient. Retention rates of enzymatic activity of irradiated preparations at the dose of 25 kGy, which is a complete sterilization dose of pharmaceutical materials, were estimated to be 83% for raw pepsin, and 86% and 93% for saccharated pepsin preparations. At the dose of 10 kGy suggested for food irradiation the retention rates were more than 93% for all pepsins. Therefore, this method is applicable considering the stability of the enzymatic activity after irradiation in the proper range of dose. However, it is necessary to consider the fact that radiolytic products of lactose affect the measurement of enzymatic activity. (author)

  16. The variations of enzymatic activity of pepsin preparation by [gamma]-irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kimura, Syojiro; Taimatsu, Meiko (Osaka Univ. of Pharmaceutical Science, Matsubara (Japan)); Kanbashi, Toshitaka; Okamoto, Shinichi; Ohnishi, Tokuhiro

    1993-02-01

    Effect of [gamma]-irradiation on the enzymatic activity of raw pepsin and some saccharated pepsin preparations were studied in the dose range from 0 to 300 kGy. As a result, the apparent reduction rate of saccharated pepsin preparations is less than of raw pepsin. K values of raw and saccharated pepsins were 0.014 and 0.0040-0.0061, and G values of raw and saccharated pepsins were 3.98 and 1.13-1.73, respectively. The lower K and G values of saccharated pepsin than those of raw pepsin seem to be due to radiolytic products of lactose in the preparations as an excipient. Retention rates of enzymatic activity of irradiated preparations at the dose of 25 kGy, which is a complete sterilization dose of pharmaceutical materials, were estimated to be 83% for raw pepsin, and 86% and 93% for saccharated pepsin preparations. At the dose of 10 kGy suggested for food irradiation the retention rates were more than 93% for all pepsins. Therefore, this method is applicable considering the stability of the enzymatic activity after irradiation in the proper range of dose. However, it is necessary to consider the fact that radiolytic products of lactose affect the measurement of enzymatic activity. (author).

  17. Enzymatic Hydrolysis of Oleuropein from Olea europea (Olive Leaf Extract and Antioxidant Activities

    Directory of Open Access Journals (Sweden)

    Jiao-Jiao Yuan

    2015-02-01

    Full Text Available Oleuropein (OE, the main polyphenol in olive leaf extract, is likely to decompose into hydroxytyrosol (HT and elenolic acid under the action of light, acid, base, high temperature. In the enzymatic process, the content of OE in olive leaf extract and enzyme are key factors that affect the yield of HT. A selective enzyme was screened from among 10 enzymes with a high OE degradation rate. A single factor (pH, temperature, time, enzyme quantity optimization process and a Box-Behnken design were studied for the enzymatic hydrolysis of 81.04% OE olive leaf extract. Additionally, enzymatic hydrolysis results with different substrates (38.6% and 81.04% OE were compared and the DPPH antioxidant properties were also evaluated. The result showed that the performance of hydrolysis treatments was best using hemicellulase as a bio-catalyst, and the high purity of OE in olive extract was beneficial to biotransform OE into HT. The optimal enzymatic conditions for achieving a maximal yield of HT content obtained by the regression were as follows: pH 5, temperature 55 °C and enzyme quantity 55 mg. The experimental result was 11.31% ± 0.15%, and the degradation rate of OE was 98.54%. From the present investigation of the antioxidant activity determined by the DPPH method, the phenol content and radical scavenging effect were both decreased after enzymatic hydrolysis by hemicellulase. However, a high antioxidant activity of the ethyl acetate extract enzymatic hydrolysate (IC50 = 41.82 μg/mL was demonstated. The results presented in this work suggested that hemicellulase has promising and attractive properties for industrial production of HT, and indicated that HT might be a valuable biological component for use in pharmaceutical products and functional foods.

  18. A specific method for measurement of nitric oxide synthase enzymatic activity in peritoneal biopsies.

    OpenAIRE

    Combet, S.; Balligand, Jean-Luc; Lameire, N.; Goffin, Eric; Devuyst, Olivier

    2000-01-01

    A specific method for measurement of nitric oxide synthase enzymatic activity in peritoneal biopsies. BACKGROUND: Nitric oxide (NO) is synthesized by NO synthase (NOS) isoforms that are expressed in the peritoneum. Thus far, NOS activity in the peritoneum has been assessed by nonspecific methods. We describe the application of a specific method for determination of NOS activity in rat and human peritoneal biopsies. METHODS: The L-citrulline assay is based on the stoechiometric production of N...

  19. Enzymatic Regulation of Steroidogenesis and Nuclear Receptor Activation : Special Focus on Vitamin D and Sex Hormones

    OpenAIRE

    LUNDQVIST, JOHAN

    2011-01-01

    Enzyme-catalyzed reactions are important to regulate steroidogenesis and nuclear receptor activation. The present investigation examines the role of steroid metabolism catalyzed by CYP7B1 for regulation of hormone receptor activation and the effects of vitamin D on enzymatic regulation of steroidogenesis. The study reports data indicating that CYP7B1 can regulate estrogenic signaling by converting estrogens into inactive or less active metabolites. Similar results were obtained for CYP7B1-med...

  20. Effect of restricted motion in high temperature on enzymatic activity of the pancreas

    Science.gov (United States)

    Abdusattarov, A.; Smirnova, G. I.

    1980-01-01

    Effects of 30 day hypodynamia coupled with high temperature (35-36 C) on enzymatic activity of the pancreas of male adult rats were studied. The test animals were divided into four groups. Group one served as controls (freedom of movement and a temperature of 25-26 C, considered optimal). The remaining animals were divided into three additional groups: Group two freedom of movement but high temperature (35-36 C); group three hypodynamia but an optimal temperature; group four hypodynamia and 35-36 C. Considerable change in the enzymatic activity in the pancreas of the four groups is observed in three experimental groups (two, three, and four) as compared to the control (group one). The results indicate that adaption of the organism to the thermal factor and restricted movement is accompanied by a change in the enzymatic spectrum of the pancreas. With the combined effect of these two stresses under conditions of the adaption of the organism especially sharp shifts occur in the enzymatic activity.

  1. Surfactant-activated lipase hybrid nanoflowers with enhanced enzymatic performance

    Science.gov (United States)

    Cui, Jiandong; Zhao, Yamin; Liu, Ronglin; Zhong, Cheng; Jia, Shiru

    2016-01-01

    Increasing numbers of materials have been extensively used as platforms for enzyme immobilization to improve catalytic performance. However, activity of the most of the enzymes was declined after immobilization. Here, we develop a surfactant-activated lipase-inorganic flowerlike hybrid nanomaterials with rational design based on interfacial activation and self-assembly. The resulting surfactant-activated lipase-inorganic hybird nanoflower (activated hNF-lipase) exhibited 460% and 200% higher activity than native lipase and conventional lipase-inorganic hybird nanoflower (hNF-lipase). Furthermore, the activated hNF-lipase displayed good reusability due to its monodispersity and mechanical properties, and had excellent long-time stability. The superior catalytic performances were attributed to both the conformational modulation of surfactants and hierarchical structure of nanoflowers, which not only anchored lipases in an active form, but also decreased the enzyme-support negative interaction and mass-transfer limitations. This new biocatalytic system is promising to find widespread use in applications related to biomedicine, biosensor, and biodiesel. PMID:27297609

  2. WATER STRESS RESPONSE ON THE ENZYMATIC ACTIVITY IN COWPEA NODULES

    Directory of Open Access Journals (Sweden)

    Figueiredo Márcia do Vale B.

    2001-01-01

    Full Text Available A greenhouse experiment was carried out aiming to study the effect of water stress on metabolic activity of cowpea nodules at different plant development stages. Cowpea plants were grown in pots with yellow latosol soil under three different matric potentials treatments: -7.0 (control-S1, -70.0 (S2 and <-85.0 KPa (S3. The experimental design was randomized blocks with sub-divided plots, each plot containing a different degree of water stress, divided in sub-plots for the four different developmental stages: E1 (0-15, E2 (15-30, E3 (20-35 and E4 (30-45 days after emmergence. Water stress treatments were applied by monitoring soil water potential using a set of porous cups. The effect of water stress was most harmful to cowpea when it was applied at E2 than at other symbiotic process stages. Shoot/root ratio decreased from 2.61 to 2.14 when matric potential treatment was <-85.0 and -70.0 KPa respectively. There was a reduction in the glutamine synthetase activity and phosphoenolpyruvate carboxilase activity with increased stress, while glutamine synthase activity was the enzyme most sensitive to water stress. Glutamate dehydrogenase activity increased in more negative matric potential, indicating that this enzyme is sufficiently activitye under water stress.

  3. Modulation of Network Activity in Dissociated Hippocampal Cultures by Enzymatic Digestion of Extracellular Matrix

    OpenAIRE

    Mukhina I.V.; Vedunova М.V.; Sakharnova Т.А.; Dityatev А.E.

    2012-01-01

    To investigate the role of extracellular matrix in spontaneous neuronal network activity, we used microelectrode array technology and enzymatic treatment of hippocampal culture with hyaluronidase, which digests the major component of extracellular matrix, hyaluronic acid. Studies were performed using hippocampal cells that were dissociated from embryonic С57ВL6 mice (E18) and plated on microelectrode arrays (MEAs). Our findings revealed that hyaluronidase promoted seizure-like activity during...

  4. Enzymatic assay for calmodulins based on plant NAD kinase activity

    Energy Technology Data Exchange (ETDEWEB)

    Harmon, A.C.; Jarrett, H.W.; Cormier, M.J.

    1984-01-01

    NAD kinase with increased sensitivity to calmodulin was purified from pea seedlings (Pisum sativum L., Willet Wonder). Assays for calmodulin based on the activities of NAD kinase, bovine brain cyclic nucleotide phosphodiesterase, and human erythrocyte Ca/sup 2 -/-ATPase were compared for their sensitivities to calmodulin and for their abilities to discriminate between calmodulins from different sources. The activities of the three enzymes were determined in the presence of various concentrations of calmodulins from human erythrocyte, bovine brain, sea pansy (Renilla reniformis), mung bean seed (Vigna radiata L. Wilczek), mushroom (Agaricus bisporus), and Tetrahymena pyriformis. The concentrations of calmodulin required for 50% activation of the NAD kinase (K/sub 0.5/) ranged from 0.520 ng/ml for Tetrahymena to 2.20 ng/ml for bovine brain. The A/sub 0.5/ s ranged from 19.6 ng/ml for bovine brain calmodulin to 73.5 ng/ml for mushroom calmodulin for phosphodiesterase activation. The K/sub 0.5/'s for the activation of Ca/sup 2 +/-ATPase ranged from 36.3 ng/mol for erythrocyte calmodulin to 61.7 ng/ml for mushroom calmodulin. NAD kinase was not stimulated by phosphatidylcholine, phosphatidylserine, cardiolipin, or palmitoleic acid in the absence or presence of Ca/sup 2 +/. Palmitic acid had a slightly stimulatory effect in the presence of Ca/sup 2 +/ (10% of maximum), but no effect in the absence of Ca/sup 2 +/. Palmitoleic acid inhibited the calmodulin-stimulated activity by 50%. Both the NAD kinase assay and radioimmunoassay were able to detect calmodulin in extracts containing low concentrations of calmodulin. Estimates of calmodulin contents of crude homogenates determined by the NAD kinase assay were consistent with amounts obtained by various purification procedures. 30 references, 1 figure, 4 tables.

  5. WATER STRESS RESPONSE ON THE ENZYMATIC ACTIVITY IN COWPEA NODULES

    OpenAIRE

    Figueiredo Márcia do Vale B.; Bezerra-Neto Egídio; Burity Hélio A.

    2001-01-01

    A greenhouse experiment was carried out aiming to study the effect of water stress on metabolic activity of cowpea nodules at different plant development stages. Cowpea plants were grown in pots with yellow latosol soil under three different matric potentials treatments: -7.0 (control-S1), -70.0 (S2) and

  6. Measuring In Vitro ATPase Activity for Enzymatic Characterization.

    Science.gov (United States)

    Rule, Chelsea S; Patrick, Marcella; Sandkvist, Maria

    2016-01-01

    Adenosine triphosphate-hydrolyzing enzymes, or ATPases, play a critical role in a diverse array of cellular functions. These dynamic proteins can generate energy for mechanical work, such as protein trafficking and degradation, solute transport, and cellular movements. The protocol described here is a basic assay for measuring the in vitro activity of purified ATPases for functional characterization. Proteins hydrolyze ATP in a reaction that results in inorganic phosphate release, and the amount of phosphate liberated is then quantitated using a colorimetric assay. This highly adaptable protocol can be adjusted to measure ATPase activity in kinetic or endpoint assays. A representative protocol is provided here based on the activity and requirements of EpsE, the AAA+ ATPase involved in Type II Secretion in the bacterium Vibrio cholerae. The amount of purified protein needed to measure activity, length of the assay and the timing and number of sampling intervals, buffer and salt composition, temperature, co-factors, stimulants (if any), etc. may vary from those described here, and thus some optimization may be necessary. This protocol provides a basic framework for characterizing ATPases and can be performed quickly and easily adjusted as necessary. PMID:27584824

  7. Enzymatic activation of oxygen in the biosynthesis of polyketide antibiotics

    OpenAIRE

    Koskiniemi, Hanna

    2009-01-01

    The electron structure of molecular oxygen (O2) is such that it cannot spontaneously react with organic molecules, a feature that protects organisms from oxidative stress. However, oxygenases can insert O2 in organic substrates. The enzymes do this after activating O2 for the reaction, most often with the aid of an organic or inorganic cofactor. A number of different cofactors have been discovered in nature thus far. In this thesis, five oxygenases with three different cata...

  8. Modelling the Effects of Ageing Time of Starch on the Enzymatic Activity of Three Amylolytic Enzymes

    Directory of Open Access Journals (Sweden)

    Nelson P. Guerra

    2012-01-01

    Full Text Available The effect of increasing ageing time (t of starch on the activity of three amylolytic enzymes (Termamyl, San Super, and BAN was investigated. Although all the enzymatic reactions follow michaelian kinetics, vmax decreased significantly (P<0.05 and KM increased (although not always significantly with the increase in t. The conformational changes produced in the starch chains as a consequence of the ageing seemed to affect negatively the diffusivity of the starch to the active site of the enzymes and the release of the reaction products to the medium. A similar effect was observed when the enzymatic reactions were carried out with unaged starches supplemented with different concentrations of gelatine [G]. The inhibition in the amylolytic activities was best mathematically described by using three modified forms of the Michaelis-Menten model, which included a term to consider, respectively, the linear, exponential, and hyperbolic inhibitory effects of t and [G].

  9. Enzymatic activity in the presence of surfactants commonly used in dissolution media, Part 1: Pepsin

    Science.gov (United States)

    Guzman, Maria L; Marques, Margareth R; Olivera ME, Maria E; Stippler, Erika S

    2016-01-01

    The United States Pharmacopeia (USP) General Chapters Dissolution 〈711〉 and Disintegration and Dissolution of Dietary Supplements 〈2040〉 allows the use of enzymes in dissolution media when gelatin capsules do not conform to dissolution specifications due to cross linking. Possible interactions between enzymes and surfactants when used together in dissolution media could result in loss of the enzymatic activity. Pepsin is an enzyme commonly used in dissolution media, and in this work, the activity of pepsin was determined in the presence of different surfactants as usually found in case of dissolution tests of certain gelatin capsule formulations. Pepsin enzymatic activity was determined according to the Ninth Edition of the Food Chemicals Codex (FCC) 9 method, in dissolution conditions: simulated gastric fluid, 37 °C and 50 rpm. Sodium dodecyl sulfate (SDS), cetyltrimethyl ammonium bromide (CTAB), polysorbate 80 (Tween 80) and octoxynol 9 (Triton X100) in concentrations above and below their critical micellar concentrations were selected. Results showed a significant reduction in the activity of pepsin at all the concentrations of SDS assayed. On the contrary, CTAB, Tween 80, and Triton X100 did not alter the enzymatic activity at of pepsin any of the concentration assayed. This data demonstrates a rational selection of the surfactant to be used when pepsin is required in dissolution test. PMID:27047734

  10. Enzymatic activity in the presence of surfactants commonly used in dissolution media, Part 1: Pepsin.

    Science.gov (United States)

    Guzman, Maria L; Marques, Margareth R; Olivera Me, Maria E; Stippler, Erika S

    2016-01-01

    The United States Pharmacopeia (USP) General Chapters Dissolution 〈711〉 and Disintegration and Dissolution of Dietary Supplements 〈2040〉 allows the use of enzymes in dissolution media when gelatin capsules do not conform to dissolution specifications due to cross linking. Possible interactions between enzymes and surfactants when used together in dissolution media could result in loss of the enzymatic activity. Pepsin is an enzyme commonly used in dissolution media, and in this work, the activity of pepsin was determined in the presence of different surfactants as usually found in case of dissolution tests of certain gelatin capsule formulations. Pepsin enzymatic activity was determined according to the Ninth Edition of the Food Chemicals Codex (FCC) 9 method, in dissolution conditions: simulated gastric fluid, 37 °C and 50 rpm. Sodium dodecyl sulfate (SDS), cetyltrimethyl ammonium bromide (CTAB), polysorbate 80 (Tween 80) and octoxynol 9 (Triton X100) in concentrations above and below their critical micellar concentrations were selected. Results showed a significant reduction in the activity of pepsin at all the concentrations of SDS assayed. On the contrary, CTAB, Tween 80, and Triton X100 did not alter the enzymatic activity at of pepsin any of the concentration assayed. This data demonstrates a rational selection of the surfactant to be used when pepsin is required in dissolution test. PMID:27047734

  11. Digestive enzymatic activity during ontogenetic development in zebrafish (Danio rerio).

    Science.gov (United States)

    Guerrera, Maria Cristina; De Pasquale, Francesca; Muglia, Ugo; Caruso, Gabriella

    2015-12-01

    Despite the growing importance of zebrafish (Danio rerio) as an experimental model in biomedical research, some aspect of physiological and related morphological age dependent changes in digestive system during larval development are still unknown. In this paper, a biochemical and morphological study of the digestive tract of zebrafish was undertaken to record the functional changes occurring in this species during its ontogenetic development, particularly from 24 hr to 47 days post fertilization (dpf). Endo- and exo-proteases, as well as α-amylase enzymes, were quantified in zebrafish larvae before first feeding (7 dpf). The most morphologically significant events during the ontogenesis of the gut occurred between 3 dpf (mouth opening) and 7 dpf (end of exocrine pancreas differentiation). The presence of a wide range of digestive enzymes, already active at earlier zebrafish larval stages, closely related with the omnivorous diet of this species. Increasing enzyme activities were found with increasing age, probably in relation with intestinal mucosa folding and consequent absorption surface increase. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 699-706, 2015. © 2015 Wiley Periodicals, Inc. PMID:26477613

  12. Effect of irradiation on APN/CD13 enzymatic activity on AGM stromal cells

    International Nuclear Information System (INIS)

    Objective: To investigate the expression of APN/CD13 in mice embryo AGM stromal cells and the change of its enzymatic activity before and after irradiation. Methods: The expression of APN/CD13 in AGM stromal cells was assayed by RT-PCR and immunohistochemistry. The stromal cells in AGM region were irradiated with 8.0 Gy of 60Co γ-rays, and APN/CD13 enzymatic activity was measured by spectrophotometer at different time points. Results: The APN/CD13 expression level was enhanced significantly in AGM stromal cells. The enzymatic activity of APN/CD13 decreased temporally post-irradiation injury, then increased to the highest level 4 hours post-irradiation, and it returned to the level of before irradiation 24 to 48 hours post-irradiation. Conclusions: The expression of APN/CD13 was remarkable in AGM stromal cells. The enzyme activity of APN/CD13 was temporally enhanced after irradiation, which might be one of the compensatory mechanisms to promote the hematopoietic recovery after irradiation. (authors)

  13. Polymerase/DNA interactions and enzymatic activity: multi-parameter analysis with electro-switchable biosurfaces

    Science.gov (United States)

    Langer, Andreas; Schräml, Michael; Strasser, Ralf; Daub, Herwin; Myers, Thomas; Heindl, Dieter; Rant, Ulrich

    2015-07-01

    The engineering of high-performance enzymes for future sequencing and PCR technologies as well as the development of many anticancer drugs requires a detailed analysis of DNA/RNA synthesis processes. However, due to the complex molecular interplay involved, real-time methodologies have not been available to obtain comprehensive information on both binding parameters and enzymatic activities. Here we introduce a chip-based method to investigate polymerases and their interactions with nucleic acids, which employs an electrical actuation of DNA templates on microelectrodes. Two measurement modes track both the dynamics of the induced switching process and the DNA extension simultaneously to quantitate binding kinetics, dissociation constants and thermodynamic energies. The high sensitivity of the method reveals previously unidentified tight binding states for Taq and Pol I (KF) DNA polymerases. Furthermore, the incorporation of label-free nucleotides can be followed in real-time and changes in the DNA polymerase conformation (finger closing) during enzymatic activity are observable.

  14. Enzymatic activity in the presence of surfactants commonly used in dissolution media, Part 1: Pepsin

    OpenAIRE

    Guzman, Maria L; Marques, Margareth R; Olivera ME, Maria E; Stippler, Erika S.

    2016-01-01

    The United States Pharmacopeia (USP) General Chapters Dissolution 〈711〉 and Disintegration and Dissolution of Dietary Supplements 〈2040〉 allows the use of enzymes in dissolution media when gelatin capsules do not conform to dissolution specifications due to cross linking. Possible interactions between enzymes and surfactants when used together in dissolution media could result in loss of the enzymatic activity. Pepsin is an enzyme commonly used in dissolution media, and in this work, the acti...

  15. The subunit composition of human extracellular superoxide dismutase (EC-SOD regulate enzymatic activity

    Directory of Open Access Journals (Sweden)

    Chr Nielsen Niels

    2007-10-01

    Full Text Available Abstract Background Human extracellular superoxide dismutase (EC-SOD is a tetrameric metalloenzyme responsible for the removal of superoxide anions from the extracellular space. We have previously shown that the EC-SOD subunit exists in two distinct folding variants based on differences in the disulfide bridge pattern (Petersen SV, Oury TD, Valnickova Z, Thøgersen IB, Højrup P, Crapo JD, Enghild JJ. Proc Natl Acad Sci USA. 2003;100(24:13875–80. One variant is enzymatically active (aEC-SOD while the other is inactive (iEC-SOD. The EC-SOD subunits are associated into covalently linked dimers through an inter-subunit disulfide bridge creating the theoretical possibility of 3 dimers (aa, ai or ii with different antioxidant potentials. We have analyzed the quaternary structure of the endogenous EC-SOD disulfide-linked dimer to investigate if these dimers in fact exist. Results The analyses of EC-SOD purified from human tissue show that all three dimer combinations exist including two homo-dimers (aa and ii and a hetero-dimer (ai. Because EC-SOD is a tetramer the dimers may combine to generate 5 different mature EC-SOD molecules where the specific activity of each molecule is determined by the ratio of aEC-SOD and iEC-SOD subunits. Conclusion This finding shows that the aEC-SOD and iEC-SOD subunits combine in all 3 possible ways supporting the presence of tetrameric enzymes with variable enzymatic activity. This variation in enzymatic potency may regulate the antioxidant level in the extracellular space and represent a novel way of modulating enzymatic activity.

  16. Enzymatic activities in traps of four aquatic species of the carnivorous genus Utricularia

    Czech Academy of Sciences Publication Activity Database

    Sirová, D.; Adamec, Lubomír; Vrba, Jaroslav

    2003-01-01

    Roč. 159, - (2003), s. 669-675. ISSN 0028-646X R&D Projects: GA AV ČR IAA6017202; GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z6017912; CEZ:AV0Z6005908 Keywords : aquatic carnivorous plants * extracellular enzymatic activity * trap fluid pH Subject RIV: EF - Botanics Impact factor: 3.118, year: 2003

  17. Rapid estimation of compost enzymatic activity by spectral analysis method combined with machine learning.

    Science.gov (United States)

    Chakraborty, Somsubhra; Das, Bhabani S; Ali, Md Nasim; Li, Bin; Sarathjith, M C; Majumdar, K; Ray, D P

    2014-03-01

    The aim of this study was to investigate the feasibility of using visible near-infrared (VisNIR) diffuse reflectance spectroscopy (DRS) as an easy, inexpensive, and rapid method to predict compost enzymatic activity, which traditionally measured by fluorescein diacetate hydrolysis (FDA-HR) assay. Compost samples representative of five different compost facilities were scanned by DRS, and the raw reflectance spectra were preprocessed using seven spectral transformations for predicting compost FDA-HR with six multivariate algorithms. Although principal component analysis for all spectral pretreatments satisfactorily identified the clusters by compost types, it could not separate different FDA contents. Furthermore, the artificial neural network multilayer perceptron (residual prediction deviation=3.2, validation r(2)=0.91 and RMSE=13.38 μg g(-1) h(-1)) outperformed other multivariate models to capture the highly non-linear relationships between compost enzymatic activity and VisNIR reflectance spectra after Savitzky-Golay first derivative pretreatment. This work demonstrates the efficiency of VisNIR DRS for predicting compost enzymatic as well as microbial activity. PMID:24398221

  18. Fusion-Triggered Switching of Enzymatic Activity on an Artificial Cell Membrane

    Directory of Open Access Journals (Sweden)

    Jun-ichi Kikuchi

    2012-05-01

    Full Text Available A nanosensory membrane device was constructed for detecting liposome fusion through changes in an enzymatic activity. Inspired by a biological signal transduction system, the device design involved functionalized liposomal membranes prepared by self-assembly of the following molecular components: a synthetic peptide lipid and a phospholipid as matrix membrane components, a Schiff’s base of pyridoxal 5’-phosphate with phosphatidylethanolamine as a thermo-responsive artificial receptor, NADH-dependent L-lactate dehydrogenase as a signal amplifier, and Cu2+ ion as a signal mediator between the receptor and enzyme. The enzymatic activity of the membrane device was adjustable by changing the matrix lipid composition, reflecting the thermotropic phase transition behavior of the lipid membranes, which in turn controlled receptor binding affinity toward the enzyme-inhibiting mediator species. When an effective fusogen anionic polymer was added to these cationic liposomes, membrane fusion occurred, and the functionalized liposomal membranes responded with changes in enzymatic activity, thus serving as an effective nanosensory device for liposome fusion detection.

  19. DNA-fueled molecular machine for label-free and non-enzymatic ultrasensitive detection of telomerase activity.

    Science.gov (United States)

    Sun, Panpan; Ran, Xiang; Liu, Chaoqun; Liu, Chaoying; Pu, Fang; Ren, Jinsong; Qu, Xiaogang

    2016-08-01

    Herein, a non-enzymatic and label-free strategy based on DNA-fueled molecular machine was developed for ultrasensitive detection of telomerase activity in cancer cell extracts even at the single-cell level. PMID:27405851

  20. The dual effects of Maillard reaction and enzymatic hydrolysis on the antioxidant activity of milk proteins.

    Science.gov (United States)

    Oh, N S; Lee, H A; Lee, J Y; Joung, J Y; Lee, K B; Kim, Y; Lee, K W; Kim, S H

    2013-08-01

    The objective of this study was to determine the enhanced effects on the biological characteristics and antioxidant activity of milk proteins by the combination of the Maillard reaction and enzymatic hydrolysis. Maillard reaction products were obtained from milk protein preparations, such as whey protein concentrates and sodium caseinate with lactose, by heating at 55°C for 7 d in sodium phosphate buffer (pH 7.4). The Maillard reaction products, along with untreated milk proteins as controls, were hydrolyzed for 0 to 3h with commercial proteases Alcalase, Neutrase, Protamex, and Flavorzyme (Novozymes, Bagsværd, Denmark). The antioxidant activity of hydrolyzed Maillard reaction products was determined by reaction with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, their 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and the ability to reduce ferric ions. Further characteristics were evaluated by the o-phthaldialdehyde method and sodium dodecyl sulfate-PAGE. The degree of hydrolysis gradually increased in a time-dependent manner, with the Alcalase-treated Maillard reaction products being the most highly hydrolyzed. Radical scavenging activities and reducing ability of hydrolyzed Maillard reaction products increased with increasing hydrolysis time. The combined products of enzymatic hydrolysis and Maillard reaction showed significantly greater antioxidant activity than did hydrolysates or Maillard reaction products alone. The hydrolyzed Maillard reaction products generated by Alcalase showed significantly higher antioxidant activity when compared with the other protease products and the antioxidant activity was higher for the whey protein concentrate groups than for the sodium caseinate groups. These findings indicate that Maillard reaction products, coupled with enzymatic hydrolysis, could act as potential antioxidants in the pharmaceutical, food, and dairy industries. PMID:23769366

  1. Yeasts from sub-Antarctic region: biodiversity, enzymatic activities and their potential as oleaginous microorganisms.

    Science.gov (United States)

    Martinez, A; Cavello, I; Garmendia, G; Rufo, C; Cavalitto, S; Vero, S

    2016-09-01

    Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures. Esterase activity was the most common enzymatic activity expressed by the yeast isolates regardless the assay temperature and inulinase was the second most common enzymatic activity. No cellulolytic activity was detected. One yeast identified as Guehomyces pullulans (8E) showed significant activity across six of seven enzymes types tested. Twenty-eight yeast isolates were classified as oleaginous, being the isolate 8E the strain that accumulated the highest levels of saponifiable lipids (42 %). PMID:27469174

  2. Methods for determining enzymatic activity comprising heating and agitation of closed volumes

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David Neil; Henriksen, Emily DeCrescenzo; Reed, David William; Jensen, Jill Renee

    2016-03-15

    Methods for determining thermophilic enzymatic activity include heating a substrate solution in a plurality of closed volumes to a predetermined reaction temperature. Without opening the closed volumes, at least one enzyme is added, substantially simultaneously, to the closed volumes. At the predetermined reaction temperature, the closed volumes are agitated and then the activity of the at least one enzyme is determined. The methods are conducive for characterizing enzymes of high-temperature reactions, with insoluble substrates, with substrates and enzymes that do not readily intermix, and with low volumes of substrate and enzyme. Systems for characterizing the enzymes are also disclosed.

  3. Blocked Enzymatic Etching of Gold Nanorods: Application to Colorimetric Detection of Acetylcholinesterase Activity and Its Inhibitors.

    Science.gov (United States)

    Saa, Laura; Grinyte, Ruta; Sánchez-Iglesias, Ana; Liz-Marzán, Luis M; Pavlov, Valeri

    2016-05-01

    The anisotropic morphology of gold nanorods (AuNRs) has been shown to lead to nonuniform ligand distribution and preferential etching through their tips. We have recently demonstrated that this effect can be achieved by biocatalytic oxidation with hydrogen peroxide, catalyzed by the enzyme horseradish peroxidase (HRP). We report here that modification of AuNRs with thiol-containing organic molecules such as glutathione and thiocholine hinders enzymatic AuNR etching. Higher concentrations of thiol-containing molecules in the reaction mixture gradually decrease the rate of enzymatic etching, which can be monitored by UV-vis spectroscopy through changes in the AuNR longitudinal plasmon band. This effect can be applied to develop novel optical assays for acetylcholinesterase (AChE) activity. The biocatalytic hydrolysis of acetylthiocholine by AChE yields thiocholine, which prevents enzymatic AuNR etching in the presence of HRP. Additionally, the same bioassay can be used for the detection of nanomolar concentrations of AChE inhibitors such as paraoxon and galanthamine. PMID:27070402

  4. Decipher the dynamic coordination between enzymatic activity and structural modulation at focal adhesions in living cells

    Science.gov (United States)

    Lu, Shaoying; Seong, Jihye; Wang, Yi; Chang, Shiou-Chi; Eichorst, John Paul; Ouyang, Mingxing; Li, Julie Y.-S.; Chien, Shu; Wang, Yingxiao

    2014-07-01

    Focal adhesions (FAs) are dynamic subcellular structures crucial for cell adhesion, migration and differentiation. It remains an enigma how enzymatic activities in these local complexes regulate their structural remodeling in live cells. Utilizing biosensors based on fluorescence resonance energy transfer (FRET), we developed a correlative FRET imaging microscopy (CFIM) approach to quantitatively analyze the subcellular coordination between the enzymatic Src activation and the structural FA disassembly. CFIM reveals that the Src kinase activity only within the microdomain of lipid rafts at the plasma membrane is coupled with FA dynamics. FA disassembly at cell periphery was linearly dependent on this raft-localized Src activity, although cells displayed heterogeneous levels of response to stimulation. Within lipid rafts, the time delay between Src activation and FA disassembly was 1.2 min in cells seeded on low fibronectin concentration ([FN]) and 4.3 min in cells on high [FN]. CFIM further showed that the level of Src-FA coupling, as well as the time delay, was regulated by cell-matrix interactions, as a tight enzyme-structure coupling occurred in FA populations mediated by integrin αvβ3, but not in those by integrin α5β1. Therefore, different FA subpopulations have distinctive regulation mechanisms between their local kinase activity and structural FA dynamics.

  5. Enzymatic activities produced by mixed Saccharomyces and non-Saccharomyces cultures: relationship with wine volatile composition.

    Science.gov (United States)

    Maturano, Yolanda Paola; Assof, Mariela; Fabani, María Paula; Nally, María Cristina; Jofré, Viviana; Rodríguez Assaf, Leticia Anahí; Toro, María Eugenia; Castellanos de Figueroa, Lucía Inés; Vazquez, Fabio

    2015-11-01

    During certain wine fermentation processes, yeasts, and mainly non-Saccharomyces strains, produce and secrete enzymes such as β-glucosidases, proteases, pectinases, xylanases and amylases. The effects of enzyme activity on the aromatic quality of wines during grape juice fermentation, using different co-inoculation strategies of non-Saccharomyces and Saccharomyces cerevisiae yeasts, were assessed in the current study. Three strains with appropriate enological performance and high enzymatic activities, BSc562 (S. cerevisiae), BDv566 (Debaryomyces vanrijiae) and BCs403 (Candida sake), were assayed in pure and mixed Saccharomyces/non-Saccharomyces cultures. β-Glucosidase, pectinase, protease, xylanase and amylase activities were quantified during fermentations. The aromatic profile of pure and mixed cultures was determined at the end of each fermentation. In mixed cultures, non-Saccharomyces species were detected until day 4-5 of the fermentation process, and highest populations were observed in MSD2 (10% S. cerevisiae/90% D. vanrijiae) and MSC1 (1% S. cerevisiae/99% C. sake). According to correlation and multivariate analysis, MSD2 presented the highest concentrations of terpenes and higher alcohols which were associated with pectinase, amylase and xylanase activities. On the other hand, MSC1 high levels of β-glucosidase, proteolytic and xylanolytic activities were correlated to esters and fatty acids. Our study contributes to a better understanding of the effect of enzymatic activities by yeasts on compound transformations that occur during wine fermentation. PMID:26386703

  6. Heterologous production, purification and characterization of enzymatically active Sindbis virus nonstructural protein nsP1.

    Science.gov (United States)

    Tomar, Shailly; Narwal, Manju; Harms, Etti; Smith, Janet L; Kuhn, Richard J

    2011-10-01

    Alphavirus nonstructural protein nsP1 possesses distinct methyltransferase (MTase) and guanylyltransferase (GTase) activities involved in the capping of viral RNAs. In alphaviruses, the methylation of GTP occurs before RNA transguanylation and nsP1 forms a covalent complex with m(7)GMP unlike the host mRNA guanylyltransferase which forms GMP-enzyme complex. In this study, full length SINV nsP1 was expressed in a soluble form with an N-terminal histidine tag in Escherichia coli and purified to homogeneity. The purified protein is enzymatically active and contains both MTase and GTase activity indicating that SINV nsP1 does not require membrane association for its enzymatic function. Biochemical analysis shows that detergents abolish nsP1 GTase activity, whereas nonionic detergents do not affect MTase activity. Furthermore, SINV nsP1 contains the metal-ion dependent GTase, whereas MTase does not require a metal ion. Circular dichroism spectroscopic analysis of purified protein indicate that nsP1 has a mixed α/β structure and is in the folded native conformation. PMID:21693190

  7. Effects of Cd and Pb pollution on soil enzymatic activities and soil microbiota

    Institute of Scientific and Technical Information of China (English)

    LIU Shuqing; YANG Zhixin; WANG Xiaomin; ZHANG Xiaogui; GAO Rutai; LIU Xia

    2007-01-01

    Based on a representative sampling method and pot experiment with different concentrations of Cd and Pd,the enzymatic activities(urease,phosphatase,catalase,invertase),population of bacteria,fungus and actinomycete in the soil,the Cd and Pd pollution status of soil samples(from the wastewater-irrigated area of Baoding suburb)were appraised.Unitary linear and nonlinear curve-fitting optimization models were applied in the research,and the relationship between Pb and Cd causing pollution and enzymatic activities of the tested soils were discussed.The research may provide a theoretical basis for protecting the environment in the region of Baiyangdian Lake,Hebei province,prevent soil pollution,and ascertain biochemical indexes,which reflect soil heavy metal pollution levels.The research results indicated that:(1)there was obvious accumulation of Pb and Cd in the wastewater-irrigated area,also the accumulation in wastewater-irrigated soil is more than that in fresh water-irrigated soil,and accumulation on surface layer was more than that in the lower layer.Pb and Cd contents in the tested soils exceeded the standards of soil background values for some major cities at home and abroad and the world soil Cd and Pb contents range.This means that the tested soil had reached a lightly polluted level;(2)there existed an obvious negative correlation between soil enzymatic activities and Pb and Cd contents in wastewaterirrigated soil,where the soil urease and catalase activities decreased obviously with the increase of Pb and Cd contents in soil.Therefore,the urease and catalase can be considered as biochemical indexes that reflect the degree of soil Pb and Cd pollution;(3)the pot experiments indicated that the influence of Cd on soil enzymatic activities was greater than that of Pb.Generally,the effect of Cd on soil phosphatase,urease,catalase is more obvious than that on invertase,while Pb has a more obvious effect on invertase than Cd;(4)pot experiments of triple cropping

  8. Radiation effects of some enzymatic activities in tissues of rats subjected to whole body gamma irradiation

    International Nuclear Information System (INIS)

    The present study deals with the changes produced in the activity of transaminases and cholinesterase in the tissues of male rats exposed to 6 Gy whole body-irradiation. The activity of these enzymes was estimated at 1, 3, 7 and 14 days following irradiation. The results indicated that radiation induced changes in the activity of glutamic oxaloacetic transaminase (GOT) and glutamic pyrovic transaminase (GPT) in liver brain and serum of white rats; as well as in the activity of liver and brain cholinesterase. Changes in the enzymatic activities are dependent on the time after irradiation and the tissue containing the enzyme. It could be concluded that each enzyme has a range of sensitivity to ionizing radiation according to its presence in the animal organ. This must serve cancer radiotherapy for patients

  9. Glycation of Ribonuclease A affects its enzymatic activity and DNA binding ability.

    Science.gov (United States)

    Dinda, Amit Kumar; Tripathy, Debi Ranjan; Dasgupta, Swagata

    2015-11-01

    Prolonged non-enzymatic glycation of proteins results in the formation of advanced glycation end products (AGEs) that cause several diseases. The glycation of Ribonuclease A (RNase A) at pH 7.4 and 37 °C with ribose, glucose and fructose has been monitored by UV-vis, fluorescence, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix assisted laser desorption ionization spectroscopy-time of flight (MALDI-TOF) methods. The enzymatic activity and DNA binding ability of glycated RNase A was also investigated by an agarose gel-based assay. A precipitation assay examined the ribonucleolytic activity of the glycated enzyme. An increase in incubation time resulted in the formation of high molecular weight AGEs with a decrease in ribonucleolytic activity. Ribose exhibits the highest potency as a glycating agent and showed the greatest reduction in the ribonucleolytic activity of the enzyme. Interestingly, glycated RNase A was unable to bind with the ribonuclease inhibitor (RI) and DNA. The glycated form of the protein was also found to be ineffective in DNA melting unlike native RNase A. PMID:26365067

  10. Gamma radiation effect on biological activity and enzymatic properties of snake venoms

    International Nuclear Information System (INIS)

    The effect of gamma radiation, from Co-60, on the biological activity and on some enzymatic activities, present in the venoms of Lachesis muta and Bothrops atrox, using samples of dried venom that had been irradiated at a dose of 0.1, 0.5 and 1.0 Mrad have been studied. Variations in the degree of hemorrhage and local necrosis were observed in albino mice injected subcutaneously with venoms of both types. The reduction of the biological activity was greater for the local hemorrhagic effect and was dependent on the doses of irradiation. The specific activity of various enzymes, present in both venoms, is affected by the gamma radiation, at a dose of 0.1 Mrad the order of increasing inactivation being: exonuclease (4%), phospholipase (24%), caseinolytic enzyme (20%), tamesterase (33%), a thrombine-like enzyme (40%), fibrinolytic enzyme (41%), 5'-nucleotidase (50%) and endonuclease (55%). The enzymatic inactivation was augmented by 0.5 and 1.0 Mrad, without maintaining an arithmetic relation. The enzyme of major resistance to the radiation was exonuclease, whereas 5'-nucleotidase and endonuclease were the most sensitive. No significant changes were observed in the spectrum of UV absorbtion (range 260 to 290 nm) nor in the contents of L-tyrosine in the irradiated venoms

  11. EPSPS variability, gene expression, and enzymatic activity in glyphosate-resistant biotypes of Digitaria insularis.

    Science.gov (United States)

    Galeano, E; Barroso, A A M; Vasconcelos, T S; López-Rubio, A; Albrecht, A J P; Victoria Filho, R; Carrer, H

    2016-01-01

    Weed resistance to herbicides is a natural phenomenon that exerts selection on individuals in a population. In Brazil, glyphosate resistance was recently detected in Digitaria insularis. The objective of this study was to elucidate mechanisms of weed resistance in this plant, including genetic variability, allelism, amino acid substitutions, gene expression, and enzymatic activity levels. Most of these have not previously been studied in this species. D. insularis DNA sequences were used to analyze genetic variability. cDNA from resistant and susceptible plants was used to identify mutations, alleles, and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) expression, using real-time quantitative reverse transcription-polymerase chain reaction. In addition, EPSPS activity was measured. We found a decrease in genetic variability between populations related to glyphosate application. Substitutions from proline to threonine and tyrosine to cysteine led to a decrease in EPSPS affinity for the glyphosate. In addition, the EPSPS enzymatic activity was slightly higher in resistant plants, whereas EPSPS gene expression was almost identical in both biotypes, suggesting feedback regulation at different levels. To conclude, our results suggest new molecular mechanisms used by D. insularis to increase glyphosate resistance. PMID:27525929

  12. Antioxidant activity and functional properties of enzymatic protein hydrolysates from common carp (Cyprinus carpio) roe (egg).

    Science.gov (United States)

    Chalamaiah, M; Jyothirmayi, T; Diwan, Prakash V; Dinesh Kumar, B

    2015-09-01

    Previously, we have reported the composition, molecular mass distribution and in vivo immunomodulatory effects of common carp roe protein hydrolysates. In the current study, antioxidative activity and functional properties of common carp (Cyprinus carpio) roe (egg) protein hydrolysates, prepared by pepsin, trypsin and Alcalase, were evaluated. The three hydrolysates showed excellent antioxidant activities in a dose dependent manner in various in vitro models such as 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azino-bis(3-ethylbenzthiazoline-6)-sulfonic acid (ABTS(+)) radical scavenging activity, ferric reducing antioxidant power (FRAP) and ferrous ion (Fe(2+)) chelating ability. Enzymatic hydrolysis significantly increased protein solubility of the hydrolysates to above 62 % over a wide pH range (2-12). Carp roe hydrolysates exhibited good foaming and emulsification properties. The results suggest that bioactive carp roe protein hydrolysates (CRPHs) with good functional properties could be useful in health food/nutraceutical/pharmaceutical industry for various applications. PMID:26344996

  13. Activity of enzymes associated with the enzymatic browning of minimally processed potatoes

    Directory of Open Access Journals (Sweden)

    Maria Carolina Dario Vitti

    2011-10-01

    Full Text Available The purpose of the present study was to evaluate the effect of different potato cultivars and storage temperatures on the specific activity of phenylalanine ammonia-lyase (PAL, polyphenol oxidase (PPO and peroxidase (POD in minimally processed potatoes. Potato cultivars Agata, Asterix and Monalisa were selected, washed, peeled, diced, sanitized, centrifuged, vacuum- packed and stored at 5 and 15°C for 9 and 5 days, respectively. There was an increase in the enzymatic activity in all the cultivars stored at 15°C. The cultivars 'Agata' and 'Asterix' stored at 5ºC did not differ significantly between them for the PAL, PPO and POD activities. The PAL, PPO and POD activities were also influenced by the storage temperature. The cultivars Agata and Asterix were more suitable in minimal processing than 'Monalisa', which was more susceptible to oxidative browning.

  14. The effects of different uranium concentrations on soil microbial populations and enzymatic activities

    International Nuclear Information System (INIS)

    Uranium is an ubiquitous constituent of natural environment with an average concentration of 4 mg/kg in earth crust. However, in local areas it may exceed the normal concentration due to human activities resulting in radionuclide contamination in groundwater and surface soil. The effect of six levels of uranium concentration (0, 50, 100,250. 500 and 1000 mg kg-1) on soil phosphatase activities and microbial populations were studied in a completely randomized design as a factorial experiment with three replications. The results showed a significant decrease in phosphatase activity. The result of the experiment suggests that soil microbial populations (bacteria, funji and actinomycetes) decrease by increasing the uranium levels in the soil. Therefore, assessment of soil enzymatic activities and microbial populations can be helpful as a useful index for a better management of uranium and radioactive contaminated soils.

  15. Selection and validation of enzymatic activities as functional markers in wood biotechnology and fungal ecology.

    Science.gov (United States)

    Mathieu, Yann; Gelhaye, Eric; Dumarçay, Stéphane; Gérardin, Philippe; Harvengt, Luc; Buée, Marc

    2013-02-15

    The dead wood and forest soils are sources of diversity and under-explored fungal strains with biotechnological potential, which require to be studied. Numerous enzymatic tests have been proposed to investigate the functional potential of the soil microbial communities or to test the functional abilities of fungal strains. Nevertheless, the diversity of these functional markers and their relevance in environmental studies or biotechnological screening does still have not been demonstrated. In this work, we assessed ten different extracellular enzymatic activities involved in the wood decaying process including β-etherase that specifically cleaves the β-aryl ether linkages in the lignin polymer. For this purpose, a collection of 26 fungal strains, distributed within three ecological groups (white, brown and soft rot fungi), has been used. Among the ten potential functional markers, the combinatorial use of only six of them allowed separation between the group of white and soft rot fungi from the brown rot fungi. Moreover, our results suggest that extracellular β-etherase is a rare and dispensable activity among the wood decay fungi. Finally, we propose that this set of markers could be useful for the analysis of fungal communities in functional and environmental studies, and for the selection of strains with biotechnological interests. PMID:23206919

  16. The influence of conformational fluctuations on enzymatic activity: modelling the functional motion of {beta}-secretase

    Energy Technology Data Exchange (ETDEWEB)

    Neri, M [SISSA and INFM, Via Beirut 2-4, I-34014 Trieste (Italy); Cascella, M [SISSA and INFM, Via Beirut 2-4, I-34014 Trieste (Italy); EPFL, Lausanne (Switzerland); Micheletti, C [SISSA and INFM, Via Beirut 2-4, I-34014 Trieste (Italy)

    2005-05-11

    Considerable insight into the functional activity of proteins and enzymes can be obtained by studying the low energy conformational distortions that the biopolymer can sustain. We carry out the characterization of these large scale structural changes for a protein of considerable pharmaceutical interest, the human {beta}-secretase. Starting from the crystallographic structure of the protein, we use the recently introduced {beta}-Gaussian model to identify, with negligible computational expenditure, the most significant distortions occurring in thermal equilibrium and the associated timescales. The application of this strategy helps us to gain considerable insight into the putative functional movements and, furthermore, allows us to identify a handful of key regions in the protein which have an important mechanical influence on the enzymatic activity despite being spatially distant from the active site. The results obtained within the Gaussian model are validated through an extensive comparison against an all-atom molecular dynamics simulation.

  17. The influence of conformational fluctuations on enzymatic activity: modelling the functional motion of β-secretase

    International Nuclear Information System (INIS)

    Considerable insight into the functional activity of proteins and enzymes can be obtained by studying the low energy conformational distortions that the biopolymer can sustain. We carry out the characterization of these large scale structural changes for a protein of considerable pharmaceutical interest, the human β-secretase. Starting from the crystallographic structure of the protein, we use the recently introduced β-Gaussian model to identify, with negligible computational expenditure, the most significant distortions occurring in thermal equilibrium and the associated timescales. The application of this strategy helps us to gain considerable insight into the putative functional movements and, furthermore, allows us to identify a handful of key regions in the protein which have an important mechanical influence on the enzymatic activity despite being spatially distant from the active site. The results obtained within the Gaussian model are validated through an extensive comparison against an all-atom molecular dynamics simulation

  18. Integrated catalysis opens new arylation pathways via regiodivergent enzymatic C–H activation

    Science.gov (United States)

    Latham, Jonathan; Henry, Jean-Marc; Sharif, Humera H.; Menon, Binuraj R. K.; Shepherd, Sarah A.; Greaney, Michael F.; Micklefield, Jason

    2016-01-01

    Despite major recent advances in C–H activation, discrimination between two similar, unactivated C–H positions is beyond the scope of current chemocatalytic methods. Here we demonstrate that integration of regioselective halogenase enzymes with Pd-catalysed cross-coupling chemistry, in one-pot reactions, successfully addresses this problem for the indole heterocycle. The resultant ‘chemobio-transformation' delivers a range of functionally diverse arylated products that are impossible to access using separate enzymatic or chemocatalytic C–H activation, under mild, aqueous conditions. This use of different biocatalysts to select different C–H positions contrasts with the prevailing substrate-control approach to the area, and presents opportunities for new pathways in C–H activation chemistry. The issues of enzyme and transition metal compatibility are overcome through membrane compartmentalization, with the optimized process requiring no intermediate work-up or purification steps. PMID:27283121

  19. Enzymatic digestive activity and absorption efficiency in Tagelus dombeii upon Alexandrium catenella exposure

    Science.gov (United States)

    Fernández-Reiriz, M. J.; Navarro, J. M.; Cisternas, B. A.; Babarro, J. M. F.; Labarta, U.

    2013-12-01

    We analyzed absorption efficiency (AE) and digestive enzyme activity (amylase, cellulase complex, and laminarinase) of the infaunal bivalve Tagelus dombeii originating from two geographic sites, Corral-Valdivia and Melinka-Aysén, which have different long-term paralytic shellfish poisoning (PSP) exposure rates. We report the effects of past feeding history (origin) on T. dombeii exposed to a mixed diet containing the toxic dinoflagellate Alexandrium catenella and another dinoflagellate-free control diet over a 12-day period in the laboratory. Absorption efficiency values of T. dombeii individuals that experienced PSP exposure in their habitat (Melinka-Aysén) remained unchanged during exposure to toxic food in the laboratory. In contrast, T. dombeii from a non-PSP exposure field site (Corral-Valdivia) showed a significant reduction in AE with toxic exposure time. This study established that the amylase and cellulase complexes were the most important enzymes in the digestive glands of Tagelus from both sites. The temporal evolution of enzymatic activity under toxic diet was fitted to exponential (amylase and cellulase) and to a logarithmic (laminarinase) models. In all fits, we found significant effect of origin in the model parameters. At the beginning of the experiment, higher enzymatic activity was observed for clams from Corral-Valdivia. The amylase activity decreased with time exposure for individuals from Corral and increased for individuals from Melinka. Cellulase activity did not vary over time for clams from Corral, but increased for individuals from Melinka and laminarinase activity decreased over time for individuals from Corral and remained unchanged over time for Melinka. A feeding history of exposure to the dinoflagellate A. catenella was reflected in the digestive responses of both T. dombeii populations.

  20. Ontogeny of nitric oxide synthase I and III protein expression and enzymatic activity in the guinea pig hippocampus.

    Science.gov (United States)

    Kimura, K A; Reynolds, J N; Brien, J F

    1999-09-01

    60. NOS enzymatic activity increased throughout prenatal and postnatal life, and attained highest activity in the adult. The developmental profile of NOS III protein expression was similar to that for NOS enzymatic activity. There was differential expression of NOS I protein, which was low in the GD 50 fetus and increased rapidly during fetal development to attain adult level by GD 62. These data suggest that the guinea pig is a reliable animal model in which to investigate the roles of NO in normal hippocampal development and in mediating neuronal injury in this brain region. PMID:10521566

  1. Evaluating the pollution from Mureş River on Arad-Pecica sector based on enzymatic activities from sediments (Western Romania)

    OpenAIRE

    Sebastian TREITLI; Mărioara Nicoleta FILIMON; Claudia PETRUCEAN

    2011-01-01

    Seven sediment samples were collected from the river Mureş from the Arad-Pecica sector and were measured the following enzymatic activities: catalase, actual and potential dehydrogenase, urease and reduction of trivalent iron (Fe3+). All this activities were detected in all analyzed samples. Based on relative values of the enzymatic activities, the enzymatic indicator of the sediment quality (EISQ) was also calculated. The lowest value of EISQ was observed in point P6 in close proximity to th...

  2. Enzymatic activity of "high-mannose" glycosylated forms of intestinal microvillar hydrolases

    DEFF Research Database (Denmark)

    Sjöström, H; Norén, Ove; Danielsen, E M

    1985-01-01

    glycosylated form, whereas no difference was observed for the two other enzymes. The change in glycosylation from high-mannose to complex form thus seems to be of importance for the enzymatic activity of sucrase-isomaltase either by direct structural involvement or by a general stabilization effect on the......The "high-mannose" glycosylated forms of aminopeptidase N (EC 3.4.11.2), maltase-glucoamylase (EC 3.2.1.20), and sucrase-isomaltase (EC 3.2.1.48, EC 3.2.1.10) have been purified. The high-mannose glycosylated form of sucrase-isomaltase was found to have a lower specific activity than the complex...

  3. AID enzymatic activity is inversely proportional to the size of cytosine C5 orbital cloud.

    Directory of Open Access Journals (Sweden)

    Gopinath Rangam

    Full Text Available Activation induced deaminase (AID deaminates cytosine to uracil, which is required for a functional humoral immune system. Previous work demonstrated, that AID also deaminates 5-methylcytosine (5 mC. Recently, a novel vertebrate modification (5-hydroxymethylcytosine - 5 hmC has been implicated in functioning in epigenetic reprogramming, yet no molecular pathway explaining the removal of 5 hmC has been identified. AID has been suggested to deaminate 5 hmC, with the 5 hmU product being repaired by base excision repair pathways back to cytosine. Here we demonstrate that AID's enzymatic activity is inversely proportional to the electron cloud size of C5-cytosine - H > F > methyl >> hydroxymethyl. This makes AID an unlikely candidate to be part of 5 hmC removal.

  4. Pertussis toxin analog with reduced enzymatic and biological activities is a protective immunogen.

    Science.gov (United States)

    Kimura, A; Mountzouros, K T; Schad, P A; Cieplak, W; Cowell, J L

    1990-01-01

    Bordetella pertussis TOX3201 has a 12-base-pair insertion in the S1 subunit gene of pertussis toxin (PTX), which encodes for a 4-amino-acid insertion between residues 107 and 108 of the mature S1 subunit (Black et al., Science 240:656-659, 1988). This mutant strain has been shown to secrete a holotoxin analog of PTX, designated CRM3201, with reduced ADP-ribosyltransferase activity. In the present study, we evaluated the biochemical, biological, and immunoprotective activities of purified CRM3201. Assay of enzymatic activities showed that CRM3201 had 20 to 30% of the ADP-ribosyltransferase activity and 55 to 60% of the NAD glycohydrolase activity of native PTX. CRM3201, however, had only 2 to 6% of the activity of PTX in clustering CHO cells, promoting leukocytosis, inducing histamine sensitization, and potentiating an anaphylactic response to bovine serum albumin. In contrast, activities associated with the B oligomer (binding to fetuin, hemagglutination of goose erythrocytes, and lymphocyte mitogen activity) were comparable to those of native PTX. Injection of BALB/c mice with CRM3201 mixed with Al(OH)3 elicited high titers of antibody to PTX (as measured by enzyme-linked immunosorbent assay), which neutralized a leukocytosis-promoting dose of PTX in these mice and neutralized PTX in a CHO cell assay. Passive transfer of the anti-CRM3201 antibody protected 20-day-old Swiss-Webster mice against a lethal aerosol challenge with B. pertussis 18323. Active immunization with CRM3201 significantly reduced lung colonization in adult BALB/c mice with a B. pertussis respiratory infection. These results demonstrate (i) that the reduced ADP-ribosyltransferase activity of CRM3201 is associated with reductions in certain biological and toxic activities of PTX (the enzymatic and biological activities are not, however, totally concordant); (ii) that CRM3201 possesses a functional B oligomer; and (iii) that CRM3201 can induce toxin-neutralizing antibodies which protect mice

  5. ENZYMATIC ACTIVITY OF SOILS EXPOSED TO TRANSPORTATION POLLUTANTS, LOCATED ALONG ROAD NO. 957

    Directory of Open Access Journals (Sweden)

    Barbara Filipek-Mazur

    2014-10-01

    Full Text Available The research was conducted in order to determine the catalase, dehydrogenase, and arylsulfatase activities of soils exposed to transportation pollutants. The research material consisted of soil samples collected from points located along road no. 957 at a section passing through Zawoja (the Malopolska Region, from places at a distance of 5 and 200 m from the road edge. The samples were collected from a 0–10 cm layer, from areas covered with grasses. No considerable diversification in the enzymatic activity of the soils, depending on their distance from the road edge, was found. The mean activity of catalase and dehydrogenases in the soils located 5 m from the road edge was, respectively, 4 and 7% greater than the activity of the soils located 200 m from the road edge. The mean arylsulfatase activity in the soils located 5 m from the road edge was 3% lower than in the soils located at a distance of 200 m. A positive correlation was found between the catalase and arylsulfatase activities, and the dehydrogenase activity in the soils.

  6. Antioxidative activities of hydrolysates from edible birds nest using enzymatic hydrolysis

    Science.gov (United States)

    Muhammad, Nurul Nadia; Babji, Abdul Salam; Ayub, Mohd Khan

    2015-09-01

    Edible bird's nest protein hydrolysates (EBN) were prepared via enzymatic hydrolysis to investigate its antioxidant activity. Two types of enzyme (alcalase and papain) were used in this study and EBN had been hydrolysed with different hydrolysis time (30, 60, 90 and 120 min). Antioxidant activities in EBN protein hydrolysate were measured using DPPH, ABTS+ and Reducing Power Assay. From this study, increased hydrolysis time from 30 min to 120 min contributed to higher DH, as shown by alcalase (40.59%) and papain (24.94%). For antioxidant assay, EBN hydrolysed with papain showed higher scavenging activity and reducing power ability compared to alcalase. The highest antioxidant activity for papain was at 120 min hydrolysis time with ABTS (54.245%), DPPH (49.78%) and Reducing Power (0.0680). Meanwhile for alcalase, the highest antioxidant activity was at 30 min hydrolysis time. Even though scavenging activity for EBN protein hydrolysates were high, the reducing power ability was quite low as compared to BHT and ascorbic Acid. This study showed that EBN protein hydrolysate with alcalase and papain treatments potentially exhibit high antioxidant activity which have not been reported before.

  7. The diversity, extracellular enzymatic activities and photoprotective compounds of yeasts isolated in Antarctica

    Directory of Open Access Journals (Sweden)

    Aline B. M Vaz

    2011-09-01

    Full Text Available The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia, Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7% produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.

  8. Evaluation of the enzymatic activity of Trichoderma inhamatum (BOL-12QD as possible biocontroller

    Directory of Open Access Journals (Sweden)

    García-Espejo Cielo Noemí

    2016-02-01

    Full Text Available It is known that Trichoderma spp. acts as a natural biocontroller of pathogen fungi, is for this reason, that this research studies the potential of its hydrolytic enzyme activity. In this article, first we determined that the speed of growth of Trichoderma inhamatum cepa BOL-12QD is 9 hours. Later, we proposed a simple and sensitive method based in the use of basal media (BM with coloidal chitin as the only carbon resource and supplemented with bromocresol purple for the qualitative determination of chitinase activi-ty. On the other hand, it was determined the celullolytic and proteolytic activities of Trichoderma in-hamatum cepa BOL-12QD and it was observed that agitation, type and concentration of sustrate are determinant factors in enzymatic production. Then, we evaluated the cellulolytic activity of Trichoderma inhamatum cepa BOL-12QD in agitation and stationary using carboxymethylcellulose (CMC as sustrate, finding that using a 2% of sustrate the highest activity is registered at 8 days of incubation in agitation with a value of 99.23 IU/L. In relation to the results at stationary the optimal value is at the fourth day with a value of 92.76 IU/L. The protease activity it was determined taking in consideration variables of agitation and stationary, using different types and concentration of sustrate at 2%, 4% y 6% (w/v of meat extract, 1%, 3%, 5% (w/v of jelly and 1%, 2%, 4% (w/v of casein. The highest protease activity was obtained at the end of the sev-enth day with an enzymatic activity of 3075.45 IU/L at stationary using a concentration of 6% (w/v of meat extract, and using jelly at 3% (w/v at stationary it was found an activity of 568.36 IU/L on the tenth day and in agitation a value of 547.27 IU/L was reached on the twelveth day, while using casein at 1% (w/v at stationary an activity of 407.06 IU/L is reached in the fifth day, and in agitation at 4% (w/v of casein a value of 547.27 IU/L is obtained on the twelfth day, while using

  9. On enzymatic activity in organic solvents as a function of enzyme history

    Energy Technology Data Exchange (ETDEWEB)

    Ke, T.; Klibanov, A.M. [Massachusetts Inst. of Tech., Cambridge, MA (United States). Dept. of Chemistry

    1998-03-20

    Catalytic activities of {alpha}-chymotrypsin and subtilisin Carlsberg in various hydrous organic solvents were measured as a function of how the enzyme suspension had been prepared. In one method, lyophilized enzyme was directly suspended in the solvent containing 1% water. In another, the enzyme was precipitated from its aqueous solution by a 100-fold dilution with an anhydrous solvent. In most cases, the reaction rate in a given nonaqueous enzymatic system strongly (up to an order of magnitude) depended on the mode of enzyme preparation. The magnitude of this dependence was markedly affected by the nature of the solvent and enzyme. A mechanistic hypothesis proposed to explain the observed dependencies was verified in additional experiments in which the water contents and enzyme history were further varied.

  10. Linking Microbial Enzymatic Activities and Functional Diversity of Soil around Earthworm Burrows and Casts.

    Science.gov (United States)

    Lipiec, Jerzy; Frąc, Magdalena; Brzezińska, Małgorzata; Turski, Marcin; Oszust, Karolina

    2016-01-01

    The aim of this work was to evaluate the effect of earthworms (Lumbricidae) on the enzymatic activity and microbial functional diversity in the burrow system [burrow wall (BW) 0-3 mm, transitional zone (TZ) 3-7 mm, bulk soil (BS) > 20 mm from the BW] and cast aggregates of a loess soil under a pear orchard. The dehydrogenase, β-glucosidase, protease, alkaline phosphomonoesterase, and acid phosphomonoesterase enzymes were assessed using standard methods. The functional diversity (catabolic potential) was assessed using the Average Well Color Development and Richness Index following the community level physiological profiling from Biolog Eco Plates. All measurements were done using soil from each compartment immediately after in situ sampling in spring. The enzymatic activites including dehydrogenase, protease, β-glucosidase and alkaline phosphomonoesterase were appreciably greater in the BW or casts than in BS and TZ. Conversely, acid phosphomonoesterase had the largest value in the BS. Average Well Color Development in both the TZ and the BS (0.98-0.94 A590 nm) were more than eight times higher than in the BWs and casts. The lowest richness index in the BS (15 utilized substrates) increased by 86-113% in all the other compartments. The PC1 in principal component analysis mainly differentiated the BWs and the TZ. Utilization of all substrate categories was the lowest in the BS. The PC2 differentiated the casts from the other compartments. The enhanced activity of a majority of the enzymes and increased microbial functional diversity in most earthworm-influenced compartments make the soils less vulnerable to degradation and thus increases the stability of ecologically relevant processes in the orchard ecosystem. PMID:27625645

  11. Effect of Direct-Current Electric Field on Enzymatic Activity and the Concentration of Laccase.

    Science.gov (United States)

    Wang, Chunxing; Zhang, Huiling; Ren, Dajun; Li, Qian; Zhang, Shuqin; Feng, Tao

    2015-09-01

    This work investigates the effect of direct-current electric field on the extracellular enzymatic activity, concentration and other experimental parameters of laccase from Trametes versicolor. The results showed that laccase could significantly contribute to the change of pH at the end of graphite electrode. In addition, it increased the electrical conductivity of the water. In the experiment, the optimum pH and catalytic pH range for laccase activity were 3.0 and pH 2.5-4.0. The application of 6 V direct current showed significant effects on the laccase enzyme activity. The activity of laccase was enhanced in the anodic region, but at the same time was strongly inhibited at the cathode. The electric charge characteristics of laccase were changed when exposed to electric field, and some laccases molecules moved to the anode, which produced a slight migration phenomenon. This study is the basis of combination of laccase and electrical technology, at the same time, providing a new direction of enhancing laccase activity. Compared to immobilization, using electric field is simple, no chemical additives, and great potential. PMID:26063937

  12. Earthworms strongly modify microbial biomass and activity triggering enzymatic activities during vermicomposting independently of the application rates of pig slurry

    Energy Technology Data Exchange (ETDEWEB)

    Aira, Manuel E-mail: aira@uvigo.es; Monroy, Fernando; Dominguez, Jorge

    2007-10-15

    We studied the relationships between earthworm activity, microbial biomass and the activation and dynamics of several enzyme activities. We carried out an experiment in which low and high rates (1.5 and 3 kg respectively) of pig slurry were applied to small scale reactors with and without earthworms. We found that extracellular enzyme activity increased with rate of pig slurry. In both rates of pig slurry applied, the presence of earthworms in young layers stimulated microbial growth which decreased once earthworms left the slurry and the layers aged. This increase was related to the initial activation of the microbial enzymes studied as correlations between microbial biomass and enzymes showed, which indicated an increase of intracellular enzyme activity. In the aged slurry, the pattern of activity of the four enzymes assayed depended on the rate of pig slurry applied. Thus, in low rate reactors, enzymatic activity through layers appeared to be related to microbial biomass, but in high rate reactors the activity of enzymes was more or less continuous. Further, these differences in overall enzyme activity agree with the variation found in extracellular enzyme activity suggesting certain dependence on substrate availability.

  13. Earthworms strongly modify microbial biomass and activity triggering enzymatic activities during vermicomposting independently of the application rates of pig slurry

    International Nuclear Information System (INIS)

    We studied the relationships between earthworm activity, microbial biomass and the activation and dynamics of several enzyme activities. We carried out an experiment in which low and high rates (1.5 and 3 kg respectively) of pig slurry were applied to small scale reactors with and without earthworms. We found that extracellular enzyme activity increased with rate of pig slurry. In both rates of pig slurry applied, the presence of earthworms in young layers stimulated microbial growth which decreased once earthworms left the slurry and the layers aged. This increase was related to the initial activation of the microbial enzymes studied as correlations between microbial biomass and enzymes showed, which indicated an increase of intracellular enzyme activity. In the aged slurry, the pattern of activity of the four enzymes assayed depended on the rate of pig slurry applied. Thus, in low rate reactors, enzymatic activity through layers appeared to be related to microbial biomass, but in high rate reactors the activity of enzymes was more or less continuous. Further, these differences in overall enzyme activity agree with the variation found in extracellular enzyme activity suggesting certain dependence on substrate availability

  14. Membrane phospholipid augments cytochrome P4501a enzymatic activity by modulating structural conformation during detoxification of xenobiotics.

    Directory of Open Access Journals (Sweden)

    Manik C Ghosh

    Full Text Available Cytochrome P450 is a superfamily of membrane-bound hemoprotein that gets involved with the degradation of xenobiotics and internal metabolites. Accumulated body of evidence indicates that phospholipids play a crucial role in determining the enzymatic activity of cytochrome P450 in the microenvironment by modulating its structure during detoxification; however, the structure-function relationship of cytochrome P4501A, a family of enzymes responsible for degrading lipophilic aromatic hydrocarbons, is still not well defined. Inducibility of cytochrome P4501A in cultured catfish hepatocytes in response to carbofuran, a widely used pesticide around the world, was studied earlier in our laboratory. In this present investigation, we observed that treating catfish with carbofuran augmented total phospholipid in the liver. We examined the role of phospholipid on the of cytochrome P4501A-marker enzyme which is known as ethoxyresorufin-O-deethylase (EROD in the context of structure and function. We purified the carbofuran-induced cytochrome P4501A protein from catfish liver. Subsequently, we examined the enzymatic activity of purified P4501A protein in the presence of phospholipid, and studied how the structure of purified protein was influenced in the phospholipid environment. Membrane phospholipid appeared to accelerate the enzymatic activity of EROD by changing its structural conformation and thus controlling the detoxification of xenobiotics. Our study revealed the missing link of how the cytochrome P450 restores its enzymatic activity by changing its structural conformation in the phospholipid microenvironment.

  15. The Enzymatic Activity of Type 1 Iodothyronine Deiodinase (D1) is Low in Liver Hemangioma: A Preliminary Study

    OpenAIRE

    Kornasiewicz, Oskar; Debski, Marcin; Stepnowska, Marta; Szałas, Anna; Bar-Andziak, Ewa; Krawczyk, Marek

    2010-01-01

    Type 1 iodothyronine deiodinase (D1) is a crucial enzyme which converts the prohormone thyroxine (T4) into active tri-iodothyronine (T3). There has been strong evidence that the metabolism of thyroid hormones is disturbed in some neoplastic tissues such as thyroid, renal, and breast cancer. However, there are few available data about D1 enzyme activity in benign tumors such as hemangioma, which is the most common primary liver tumor. Hence this study aimed to determine the enzymatic activity ...

  16. STUDY ON QUALITY PARAMETERS AND ENZYMATIC ACTIVITY OF GRAIN MILL PRODUCTS REGION IN TRANSYLVANIA

    Directory of Open Access Journals (Sweden)

    Glevitzky Mirel

    2011-07-01

    Full Text Available This paper aims at determining the main quality parameters of grain mill products in the Transylvania region, also studying and emphasizing the enzymatic activity of flour. Determination of quality characteristics of grain mill products entails establishing physical, chemical and sensory parameters and assessing them against the limits imposed by law. Analysis was performed on samples formed by mixing basic medium extracted from different batches. Incremental size, sampling tools, how to extract them, the training sample and laboratory environments, packaging and labeling of samples were performed according to STAS 1068 69. Determination of the fall (Falling Number, an empirical test that relies on the ability of endogenous ?-amylase to reduce viscosity of the treated warm flour suspension is used, large scale milling and bakery industry to predict and assess the Baking quality of flour. In sprouted wheat, characterised by a low Falling number, dextrin produced by the action of ?-amylase leads to a sticky bread core. Experiments suggest that the values fall turnover (FN does not shrink in direct proportion to the percentage of germinating seeds. Amylolytic activity depends on the stage of sprouting of grains. Lack of ?-amylase activity can be corrected by adding malt grain ?-amylase or fungal ?-amylase.

  17. Bioengineering of stainless steel surface by covalent immobilization of enzymes. Physical characterization and interfacial enzymatic activity.

    Science.gov (United States)

    Caro, Anne; Humblot, Vincent; Méthivier, Christophe; Minier, Michel; Barbes, Lucica; Li, Joachim; Salmain, Michèle; Pradier, Claire-Marie

    2010-09-01

    Two hydrolytic enzymes, namely lysozyme and trypsin, were covalently immobilized onto stainless steel surfaces using wet chemistry processes. The immobilization strategy took advantage of the spontaneous physisorption of the polymer poly(ethylene imine) (PEI) onto stainless steel to yield a firmly attached, thin organic layer containing a high density of primary amine functions. Both enzymes were then covalently grafted to the surface via a glutaraldehyde cross-linker. Alternatively, a thicker underlayer of PEI was chemisorbed by cross-linking two PEI layers by glutaraldehyde. The effective presence of both enzymes on the stainless steel surfaces and their relative amount were assessed by immunochemical assays employing specific anti-enzyme antibodies. Eventually, the hydrolytic activity of the immobilized enzymes was evaluated by local enzymatic tests with suitable substrates. This work demonstrates that, although the amount of enzymes did not vary significantly with the underlayer thickness, their hydrolytic activity could be much improved by increasing the distance from the oxide surface and, likely, by favoring their accessibility. Our data suggest that the immobilization of enzymes on solid oxide surfaces is feasible and efficient, and that the enzymes retain catalytic activity. It may thus provide a promising route towards biofilm-resistant materials. PMID:20566201

  18. Antimicrobial effect and enzymatic activity of extract of Zingiber officinale Roscoe and stability in topical preparations

    Directory of Open Access Journals (Sweden)

    Gisele Mara Silva Gonçalves

    2014-04-01

    Full Text Available The rhizomes of common ginger (Zingiber officinale Roscoe contain substances with antimicrobial activity and proteolytic enzymes and thus may have various pharmaceutical applications. The aim of the present study was to prepare Zingiber officinale Roscoe rhizome extracts for pharmaceutical use, preserving the proteolytic enzyme activity and testing the antimicrobial activity, in order to develop topical formulations. Two extracts were obtained - aqueous and glycolic – and assessed for their physical, physicochemical and organoleptic characteristics. To measure their proteolytic activity, the extracts were assayed for enzymatic hydrolysis of 1.2% casein solution, at pH 6.0 and 37°C; papain was used for comparison. The antimicrobial activity of the glycolic extract of Zingiber officinale Roscoe was tested by microdilution; the inoculants were prepared from cultures of Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. There was no growth of S. aureus and S. epidermidis at concentrations of 150 mg/mL or more of extract, whereas P. aeruginosa was inhibited from 100 mg/mL and E.coli from 75 mg/ mL. Emulsion formulations were prepared as vehicles for the extracts and their stability was tested. The results showed proteolytic activity in both Z. officinale rhizome extracts, the glycolic extract having 691.68 PU/g juice and the aqueous extract, 338.14 PU/g juice. The formulations were stable, especially the one that contained the glycolic extract. In sum, the formulations showed satisfactory stability and the Z. officinale extract showed bactericidal activity against the cultures tested; the results are promising for the use of the extract in foods, medicine and cosmetics.

  19. Formation of marine snow and enhanced enzymatic activities in oil-contaminated seawater

    Science.gov (United States)

    Ziervogel, K.; McKay, L.; Yang, T.; Rhodes, B.; Nigro, L.; Gutierrez, T.; Teske, A.; Arnosti, C.

    2010-12-01

    The fate of oil spilled into the ocean depends on its composition, as well as on biological, chemical, and physical characteristics of the spill site. We investigated the effects of oil addition from the Deepwater Horizon (DH) spill on otherwise uncontaminated water collected close to the spill site. Incubation on a roller table mimicked the physical dynamics of natural seawater, leading to the formation of marine snow-oil aggregates. We measured the enzymatic activities of heterotrophic microbes associated with the aggregates and in the surrounding water, and assessed microbial population and community composition as oil-marine snow aggregates formed and aged in the water. Surface seawater taken near the spill site in May 2010 that had no visible crude oil was incubated in 1-l glass bottles with (oil-bottles) and without (no-oil bottles) a seawater-oil mixture collected from the same site. In the oil-bottles formation of brownish, densely packed marine snow (2-3 cm diameter) was observed within the first hour of the roller table incubation. In contrast no-oil bottles showed aggregate formation only after 3 days, and aggregates were almost transparent, less abundant, and smaller in size (roller table incubation, and analyzed for bacterial abundance and community structure as well as the activities of hydrolytic enzymes that are used by heterotrophic bacteria to degrade organic matter. We monitored oil-degrading activities with MUF-stearate and -butyrate, and also measured b-glucosidase, alkaline phosphatase, aminopeptidase, and six different polysaccharide hydrolase activities. Enzymatic activities were up to one order of magnitude higher in the oil-bottles compared with the no-oil bottles throughout the entire incubation time. Butyrate hydrolysis was elevated throughout the time course of the incubation, and stearate hydrolysis was particularly high over the initial 10 days. Activities of enzymes not directly associated with metabolism of oil were also enhanced

  20. Enzymatically synthesized inorganic polymers as morphogenetically active bone scaffolds: application in regenerative medicine.

    Science.gov (United States)

    Wang, Xiaohong; Schröder, Heinz C; Müller, Werner E G

    2014-01-01

    In recent years a paradigm shift in understanding of human bone formation has occurred that starts to change current concepts in tissue engineering of bone and cartilage. New discoveries revealed that fundamental steps in biomineralization are enzyme driven, not only during hydroxyapatite deposition, but also during initial bioseed formation, involving the transient deposition and subsequent transformation of calcium carbonate to calcium phosphate mineral. The principal enzymes mediating these reactions, carbonic anhydrase and alkaline phosphatase, open novel targets for pharmacological intervention of bone diseases like osteoporosis, by applying compounds acting as potential activators of these enzymes. It is expected that these new findings will give an innovation boost for the development of scaffolds for bone repair and reconstruction, which began with the use of bioinert materials, followed by bioactive materials and now leading to functional regenerative tissue units. These new developments have become possible with the discovery of the morphogenic activity of bioinorganic polymers, biocalcit, bio-polyphosphate and biosilica that are formed by a biogenic, enzymatic mechanism, a driving force along with the development of novel rapid-prototyping three-dimensional (3D) printing methods and bioprinting (3D cell printing) techniques that may allow a fabrication of customized implants for patients suffering in bone diseases in the future. PMID:25376489

  1. Molecular characterization and enzymatic activity of laccases in two Pleurotus spp. with different pathogenic behaviour.

    Science.gov (United States)

    Punelli, Federico; Reverberi, Massimo; Porretta, Daniele; Nogarotto, Sara; Fabbri, Anna A; Fanelli, Corrado; Urbanelli, Sandra

    2009-03-01

    Pleurotus eryngii and P. ferulae, two species belonging to the P. eryngii complex, synthesize laccases, ligninolytic enzymes that play a role in the host-pathogen interaction in the first step of infection. Ecological studies have shown that although both fungi have been recognized as saprophytes, P. eryngii weakly pathogenic when colonizing the roots and stems of Eryngium campestre, whereas P. ferulae is mostly pathogenic to Ferula communis. The paper describes the genomic organization of four putative laccase genes (lac1, lac2, lac3, and lac5-like gene; gene names were assigned on the basis of sequence homologies) of P. eryngii and P. ferulae. The mRNA expression and enzymatic activity of the laccases were analysed under culture conditions where a source of lignin (wheat bran) or lyophilized roots of E. campestre or F. communis were present. These experiments indicated that the four lac-like genes were differentially regulated in the two mushrooms. Specifically, the addition of the lyophilized roots of the respective host plant to the culture media induced an advance in the mRNA expression of the four lac-like genes and a seven-fold higher total laccase activity in P. ferulae than in P. eryngii. The results obtained are discussed in relation to the possible role of laccases in the interaction of P. eryngii and P. ferulae with their respective host. PMID:19116166

  2. α-Galactosidase-A Loaded-Nanoliposomes with Enhanced Enzymatic Activity and Intracellular Penetration.

    Science.gov (United States)

    Cabrera, Ingrid; Abasolo, Ibane; Corchero, José L; Elizondo, Elisa; Gil, Pilar Rivera; Moreno, Evelyn; Faraudo, Jordi; Sala, Santi; Bueno, Dolores; González-Mira, Elisabet; Rivas, Merche; Melgarejo, Marta; Pulido, Daniel; Albericio, Fernando; Royo, Miriam; Villaverde, Antonio; García-Parajo, Maria F; Schwartz, Simó; Ventosa, Nora; Veciana, Jaume

    2016-04-01

    Lysosomal storage disorders (LSD) are caused by lysosomal dysfunction usually as a consequence of deficiency of a single enzyme required for the metabolism of macromolecules, such as lipids, glycoproteins, and mucopolysaccharides. For instance, the lack of α-galactosidase A (GLA) activity in Fabry disease patients causes the accumulation of glycosphingolipids in the vasculature leading to multiple organ pathology. Enzyme replacement therapy, which is the most common treatment of LSD, exhibits several drawbacks mainly related to the instability and low efficacy of the exogenously administered therapeutic enzyme. In this work, the unprecedented increased enzymatic activity and intracellular penetration achieved by the association of a human recombinant GLA to nanoliposomes functionalized with Arginine-Glycine-Aspartic acid (RGD) peptides is reported. Moreover, these new GLA loaded nanoliposomes lead to a higher efficacy in the reduction of the GLA substrate named globotriasylceramide in a cellular model of Fabry disease, than that achieved by the same concentration of the free enzyme. The preparation of these new liposomal formulations by DELOS-SUSP, based on the depressurization of a CO2 -expanded liquid organic solution, shows the great potential of this CO2 -based methodology for the one-step production of protein-nanoliposome conjugates as bioactive nanomaterials with therapeutic interest. PMID:26890358

  3. Green Tea and Bone Marrow Transplantation: From Antioxidant Activity to Enzymatic and Multidrug-resistance Modulation.

    Science.gov (United States)

    Peluso, Ilaria; Palmery, Maura; Vitalone, Annabella

    2016-10-25

    Epigallocatechin-3-gallate (EGCG), the main flavonoid of green tea (GT), could play an active role in the prevention of oxidative-stress-related diseases, such as hematologic malignancies. Some effects of EGCG are not imputable to antioxidant activity, but involve modulation of antioxidant enzymes and uric acid (UA) levels. The latter is the major factor responsible of the plasma non-enzymatic antioxidant capacity (NEAC). However, hyperuricemia is a frequent clinical feature caused by tumor lysis syndrome or cyclosporine side effects, both before and after bone marrow transplantation (BMT). Besides this, food-drug interactions could be associated with GT consumption and could have clinical implications. The molecular mechanisms involved in the redox and drug metabolizing/transporting pathways were discussed, with particular reference to the potential role of GT and EGCG in BMT. Moreover, on reviewing data on NEAC, isoprostanes, uric acid, and various enzymes from human studies on GT, its extract, or EGCG, an increase in NEAC, without effect on isoprostanes, and contrasting results on UA and enzymes were observed. Currently, few and contrasting available evidences suggest caution for GT consumption in BMT patients and more studies are needed to better understand the potential impact of EGCG on oxidative stress and metabolizing/transporting systems. PMID:26047551

  4. Single-molecule kinetics under force: probing protein folding and enzymatic activity with optical tweezers

    Science.gov (United States)

    Wong, Wesley

    2010-03-01

    Weak non-covalent bonds between and within single molecules govern many aspects of biological structure and function (e.g. DNA base-paring, receptor-ligand binding, protein folding, etc.) In living systems, these interactions are often subject to mechanical forces, which can greatly alter their kinetics and activity. My group develops and applies novel single-molecule manipulation techniques to explore and quantify these force-dependent kinetics. Using optical tweezers, we have quantified the force-dependent unfolding and refolding kinetics of different proteins, including the cytoskeletal protein spectrin in collaboration with E. Evans's group [1], and the A2 domain of the von Willebrand factor blood clotting protein in collaboration with T. Springer's group [2]. Furthermore, we have studied the kinetics of the ADAMTS13 enzyme acting on a single A2 domain, and have shown that physiolgical forces in the circulation can act as a cofactor for enzymatic cleavage, regulating hemostatic activity [2]. References: 1. E. Evans, K. Halvorsen, K. Kinoshita, and W.P. Wong, Handbook of Single Molecule Biophysics, P. Hinterdorfer, ed., Springer (2009). 2. X. Zhang, K. Halvorsen, C.-Z. Zhang, W.P. Wong, and T.A. Springer, Science 324 (5932), 1330-1334 (2009).

  5. Haematology, genotoxicity, enzymatic activity and histopathology as biomarkers of metal pollution in the shrew Crocidura russula

    Energy Technology Data Exchange (ETDEWEB)

    Sanchez-Chardi, A. [Departament Biologia Animal, Facultat de Biologia, Universitat Barcelona, Av. Diagonal 645, 08028 Barcelona (Spain); Servei de Microscopia, Facultat de Ciencies, Ed. C, Universitat Autonoma de Barcelona, 08193 Bellaterra, Barcelona (Spain)], E-mail: Alejandro.Sanchez.Chardi@uab.es; Marques, C.C.; Gabriel, S.I. [Centro de Biologia Ambiental, Departamento de Biologia Animal, Faculdade de Ciencias, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa (Portugal); Capela-Silva, F. [Centro de Investigacao em Ciencias e Tecnologias da Saude, Departamento de Biologia, Universidade de Evora, 7002-552 Evora (Portugal); Cabrita, A.S. [Centro de Histofisiologia, Instituto de Patologia Experimental, Faculdade de Medicina, Universidade de Coimbra, 3004-504 Coimbra (Portugal); Lopez-Fuster, M.J.; Nadal, J. [Departament Biologia Animal, Facultat de Biologia, Universitat Barcelona, Av. Diagonal 645, 08028 Barcelona (Spain); Mathias, M.L. [Centro de Biologia Ambiental, Departamento de Biologia Animal, Faculdade de Ciencias, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa (Portugal)

    2008-12-15

    Haematological (WBC, RBC, Hgb and Hct) and genotoxicity (MNT) parameters, hepatic enzymatic activities (GST, GPx and GR), and a histopathological evaluation of liver, kidneys and gonads were assessed as general biomarkers of metal pollution in the shrew Crocidura russula inhabiting a pyrite mining area. Specimens exposed to metals presented a few significant alterations when compared with reference animals: GST activity decreased; micronuclei increased; and evident liver alterations related to metal exposure were observed. On the basis of all the parameters studied, age was an important factor that partly explained the observed variation, whereas sex was the least important factor. Significant correlations were also found between heavy metal concentrations and biomarkers evaluated, demonstrating the great influence of these metals in the metabolic alterations. To the best of our knowledge, these data constitute the first measurements of a battery of biomarkers in shrews from a mine site and are among the few available for insectivorous mammals. - Metals from an abandoned pyrite mine produce alterations in haematological parameters, GST, MNT, and histopathology in shrews.

  6. Active evaluation of the immobilized enzyme with the unequal enzymatic activity distribution; Fukin'itsu koso kassei bunpu wo motsu koteika koso no kassei hyoka

    Energy Technology Data Exchange (ETDEWEB)

    Aoki, Toshihiro; Ajiri, Masafumi; Arai, Kunio

    1999-04-05

    In this study, enzymatic activity evaluation method which can be applied to the immobilization oxygen with the unequal enzymatic activity. Distribution is proposed. The enzymatic activity distribution was evaluated by taking up esterification reaction by immobilization lipase (Lypozyme), and analyzing the relationship between reaction rate and substrate concentration, when effective diffusion coefficient (De) and Michaelis constant (Km) in the carrier particle were independently evaluated. De was evaluated by the step response experiment using the packed bed which stocked anion exchange resin (Duolite A-568) which is a carrier of Lypozyme. It was able to be described as a result of basing response curves on adsorption of the tracer to the particle pore solid surface and mass transfer in the pore. Km was evaluated from the relationship between substrate concentration under the condition of which the effect on reaction rate is small and reaction rate of the mass transfer in the particle. In Lypozyme, the result showed that the enzymatic activity was distributed only in the carrier surface vicinity. And, it was possible to describe the immobilized enzyme reaction under the wide condition by evaluating effect and enzymatic activity distribution of the mass transfer in the particle. (translated by NEDO)

  7. Angiotensin-converting enzyme inhibitory and antioxidant activities of enzymatically synthesized phenolic and vitamin glycosides.

    Science.gov (United States)

    Charles, Rajachristu Einstein; Ponrasu, Thangavel; Sivakumar, Ramaiah; Divakar, Soundar

    2009-03-01

    Amyloglucosidase from Rhizopus mould and beta-glucosidase from sweet almond were employed for the preparation of phenolic and vitamin glycosides of vanillin, N-vanillylnonanamide, DL-dopa, dopamine, curcumin, alpha-tocopherol (vitamin E), pyridoxine (vitamin B(6)), ergocalciferol (vitamin D(2)), thiamin (vitamin B(1)) and riboflavin (vitamin B(2)). Approx. 20 enzymatically prepared phenolic and vitamin glycosides were subjected to ACE (angiotensin-converting enzyme) inhibition activity measurements, and 14 glycosides were tested for antioxidant activities. Both phenolic and vitamin glycosides exhibited IC(50) values for ACE inhibition in the 0.52+/-0.03-3.33+/-0.17 mM range and antioxidant activities ranging from 0.8+/-0.04 to 1.18+/-0.06 mM. Comparable ACE inhibition values were observed between free phenols and vitamin glycosides. However, antioxidant activities of glycosides were, in general, lesser than those of free phenols. Best IC(50) value for ACE inhibition were observed for 11-O-(D-fructofuranosyl)thiamin (0.52+/-0.03 mM), 3-hydroxy-4-O-(6-D-sorbitol)phenylalanine (0.56+/-0.03 mM), 4-O-(beta-D-glucopyranosyl)vanillin (0.61+/-0.03 mM), 4-O-(D-galactopyranosyl)vanillin (0.61+/-0.03 mM) and pyridoxine-D-glucoside (0.84+/-0.04 mM). Similarly, best IC(50) values for antioxidant activity were observed for 1,7-O-(bis-beta-D-glucopyranosyl)curcumin (0.8+/-0.04 mM), 4-O-(beta-D-glucopyranosyl)vanillin (0.9+/-0.05 mM), 3-hydroxy-4-O-(beta-D-galactopyranosyl-(1'-->4)beta-D-glucopyranosyl)phenylalanine (0.9+/-0.05 mM), 20-O-(D-glucopyranosyl)ergocalciferol (0.9+/-0.05 mM) and dopamine-D-galactoside (0.93+/-0.05 mM). PMID:18547170

  8. Enzymatic Activity of the Mycelium Compared with Oospore Development During Infection of Pea Roots by Aphanomyces euteiches

    DEFF Research Database (Denmark)

    Kjøller, Rasmus; Rosendahl, Søren

    1998-01-01

    alkaline phosphatase activity in the roots. Additionally, enzyme activity was measured after electrophoresis of an A. euteiches-specific glucose-6-phosphate isozyme. Development of oospores in the roots was measured after staining the oospores with trypan blue. In plants inoculated with the higher zoospore......To describe the disease cycle of the root pathogen Aphanomyces euteiches, enzymatic activity in the mycelium was compared with the development of oospores in pea roots. Plants were inoculated with two zoospore concentrations to achieve different disease levels. Hyphae were stained for fungal...... concentration, the enzymatic activity of the pathogen mycelium peaked 10 to 14 days after inoculation, when oospore formation was initiated. Oospore formation was associated with a gradual increase in disease symptoms. At the last harvest, plants inoculated with the higher zoospore concentration had died. In...

  9. Mapping of enzyme activity by detection of enzymatic products during AFM imaging with integrated SECM-AFM probes

    Energy Technology Data Exchange (ETDEWEB)

    Kranz, C.; Kueng, A; Lugstein, A.; Bertagnolli, E.; Mizaikoff, B

    2004-08-15

    With the integration of submicro- and nanoelectrodes into atomic force microscopy (AFM) probes using microfabrication techniques, an elegant approach combining scanning electrochemical microscopy (SECM) with AFM has recently been introduced. Simultaneous contact mode imaging of a micropatterned sample with immobilized enzyme spots and imaging of enzyme activity is shown. In contrast to force spectroscopy the conversion of an enzymatic byproduct is directly detected during AFM imaging and correlated to the activity of the enzyme.

  10. DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I

    OpenAIRE

    Birgitta Ruth Knudsen; Oskar Franch; Joanna Proszek; Yi-Ping Ho; Magnus Stougaard; Emil Laust Kristoffersen; Lærke Bay Marcussen; Morten Leth Jepsen

    2013-01-01

    Sensors capable of quantitative real-time measurements may present the easiest and most accurate way to study enzyme activities. Here we present a novel DNA-based sensor for specific and quantitative real-time measurement of the enzymatic activity of the essential human enzyme, topoisomerase I. The basic design of the sensor relies on two DNA strands that hybridize to form a hairpin structure with a fluorophore-quencher pair. The quencher moiety is released from the sensor upon reaction with ...

  11. EFFECTS OF INTERFERON THERAPY UPON IMMUNE MARKER PROFILE AND ENZYMATIC ACTIVITY IN PERIPHERAL BLOOD LYMPHOCYTES OF PATIENTS WITH RENAL CANCER

    Directory of Open Access Journals (Sweden)

    L. M. Kurtasova

    2014-01-01

    Full Text Available We have observed forty-four patients with metastatic renal cancer before and after interferon therapy. Immune markers of of peripheral blood lymphocytes were determined by flow cytometry. Activity of NAD (P-dependent dehydrogenase in blood lymphocytes was studied by means of bioluminescence technique. Changes of immune marker profiles and enzymatic activities of peripheral blood lymphocytes were found in patients with renal cancer after a course of interferon therapy.

  12. Radioimmunoassay for human pancreatic amylase: comparison of human serum amylase by measurement of enzymatic activity and by radioimmunoassay

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) for human pancreatic amylase has been developed for the determination of human serum amylase content. The assay was shown to be sensitive (7 ng/mI), reproducible and specific, but human pancreatic amylase and salivary amylase could not be distinguished by the antiserum used. In normal subjects, the mean concentration of amylase determined by the RIA was found to be 122.1 ng/ml (range: 55-250 ng/ml). A good correlation was observed between the concentration of amylase and its enzymatic activity in normal subjects. In some instances with high amylase activity, however, the rise in enzymatic activity was not accompanied by increasing amount of amylase content. (Auth.)

  13. Changes in antioxidant and antiinflammatory activity of black bean (Phaseolus vulgaris L.) protein isolates due to germination and enzymatic digestion.

    Science.gov (United States)

    López-Barrios, Lidia; Antunes-Ricardo, Marilena; Gutiérrez-Uribe, Janet A

    2016-07-15

    Germination is an inexpensive process to improve the nutritional properties of legumes. The effect of germinating black bean seeds on the production of cotyledon protein hydrolysates (CPH) with antioxidant and antiinflammatory activities was analyzed in this research. After simulated enzymatic digestion, the oxygen radical absorbance capacity (ORAC) of CPH obtained from germinated black beans was lower than that observed for raw cotyledons. There were no significant differences among CPH cellular antioxidant activities (CAA), except for the high CAA of the 120 min hydrolysate obtained from one day germinated black bean cotyledons. The most significant changes due to germination and enzymatic hydrolysis were observed for the inhibition of nitric oxide (NO) production in macrophages. The NO synthesis inhibition observed for raw CPH was reduced after simulated gastrointestinal digestion but for germinated samples the inhibition was doubled. Peptides derived from cell wall proteins produced during germination could be responsible of antiinflammatory activity. PMID:26948633

  14. Effect of combined pollution by heavy metals on soil enzymatic activities in areas polluted by tailings from Pb-Zn-Ag mine

    Institute of Scientific and Technical Information of China (English)

    CHEN Cheng-li; LIAO Min; HUANG Chang-yong

    2005-01-01

    Some enzymatic activities were determined in the areas polluted by tailings from Tiantai Pb-Zn-Ag Mine in Zhejiang Province of China. The results indicated the soil enzymatic activities decreased significantly with increase of concentrations of heavy metals or the distance away from mining tailing center, especially dehydrogenase and urease activities. Multivariate regression analysis between heavy metal contents and soil enzymatic activities indicated that single dehydrogenase activity was very significantly correlated to combined effect of soil heavy metals in mine area. Moreover, single urease, protease and acid phosphatase activities were significantly related to the combined effect of heavy metals. The results suggest it is feasible to use soil enzymatic activities to indicate the pollution situation by combined heavy metals in the soil of mine area.

  15. Characterization of Amino Acid Profile and Enzymatic Activity in Adult Rat Astrocyte Cultures.

    Science.gov (United States)

    Souza, Débora Guerini; Bellaver, Bruna; Hansel, Gisele; Arús, Bernardo Assein; Bellaver, Gabriela; Longoni, Aline; Kolling, Janaina; Wyse, Angela T S; Souza, Diogo Onofre; Quincozes-Santos, André

    2016-07-01

    Astrocytes are multitasking players in brain complexity, possessing several receptors and mechanisms to detect, participate and modulate neuronal communication. The functionality of astrocytes has been mainly unraveled through the study of primary astrocyte cultures, and recently our research group characterized a model of astrocyte cultures derived from adult Wistar rats. We, herein, aim to characterize other basal functions of these cells to explore the potential of this model for studying the adult brain. To characterize the astrocytic phenotype, we determined the presence of GFAP, GLAST and GLT 1 proteins in cells by immunofluorescence. Next, we determined the concentrations of thirteen amino acids, ATP, ADP, adenosine and calcium in astrocyte cultures, as well as the activities of Na(+)/K(+)-ATPase and acetylcholine esterase. Furthermore, we assessed the presence of the GABA transporter 1 (GAT 1) and cannabinoid receptor 1 (CB 1) in the astrocytes. Cells demonstrated the presence of glutamine, consistent with their role in the glutamate-glutamine cycle, as well as glutamate and D-serine, amino acids classically known to act as gliotransmitters. ATP was produced and released by the cells and ADP was consumed. Calcium levels were in agreement with those reported in the literature, as were the enzymatic activities measured. The presence of GAT 1 was detected, but the presence of CB 1 was not, suggesting a decreased neuroprotective capacity in adult astrocytes under in vitro conditions. Taken together, our results show cellular functionality regarding the astrocytic role in gliotransmission and neurotransmitter management since they are able to produce and release gliotransmitters and to modulate the cholinergic and GABAergic systems. PMID:26915106

  16. Activation and stabilization of the hydroperoxide lyase enzymatic extract from mint leaves (Mentha spicata) using selected chemical additives.

    Science.gov (United States)

    Akacha, Najla B; Karboune, Salwa; Gargouri, Mohamed; Kermasha, Selim

    2010-03-01

    The effects of selected lyoprotecting excipients and chemical additives on the specific activity and the thermal stability of the hydroperoxide lyase (HPL) enzymatic extract from mint leaves were investigated. The addition of KCl (5%, w/w) and dextran (2.5%, w/w) to the enzymatic extract, prior to lyophilization, increased the HPL specific activity by 2.0- and 1.2-fold, respectively, compared to the control lyophilized extract. From half-life time (t (1/2)), it can be seen that KCl has enhanced the HPL stability by 1.3- to 2.3-fold, during long-period storage at -20 degrees Celsius and 4 degrees Celsius. Among the selected additives used throughout this study, glycine appeared to be the most effective one. In addition to the activation effect conferred by glycine, it also enhanced the HPL thermal stability. In contrast, polyhydroxyl-containing additives were not effective for stabilizing the HPL enzymatic extract. On the other hand, there was no signification increase in HPL activity and its thermal stability with the presence of Triton X-100. The results also showed that in the presence of glycine (10%), the catalytic efficiency of HPL was increased by 2.45-fold than that without additive. PMID:19430937

  17. Impaired enzymatic defensive activity, mitochondrial dysfunction and proteasome activation are involved in RTT cell oxidative damage.

    Science.gov (United States)

    Cervellati, Carlo; Sticozzi, Claudia; Romani, Arianna; Belmonte, Giuseppe; De Rasmo, Domenico; Signorile, Anna; Cervellati, Franco; Milanese, Chiara; Mastroberardino, Pier Giorgio; Pecorelli, Alessandra; Savelli, Vinno; Forman, Henry J; Hayek, Joussef; Valacchi, Giuseppe

    2015-10-01

    A strong correlation between oxidative stress (OS) and Rett syndrome (RTT), a rare neurodevelopmental disorder affecting females in the 95% of the cases, has been well documented although the source of OS and the effect of a redox imbalance in this pathology has not been yet investigated. Using freshly isolated skin fibroblasts from RTT patients and healthy subjects, we have demonstrated in RTT cells high levels of H2O2 and HNE protein adducts. These findings correlated with the constitutive activation of NADPH-oxidase (NOX) and that was prevented by a NOX inhibitor and iron chelator pre-treatment, showing its direct involvement. In parallel, we demonstrated an increase in mitochondrial oxidant production, altered mitochondrial biogenesis and impaired proteasome activity in RTT samples. Further, we found that the key cellular defensive enzymes: glutathione peroxidase, superoxide dismutase and thioredoxin reductases activities were also significantly lower in RTT. Taken all together, our findings suggest that the systemic OS levels in RTT can be a consequence of both: increased endogenous oxidants as well as altered mitochondrial biogenesis with a decreased activity of defensive enzymes that leads to posttranslational oxidant protein modification and a proteasome activity impairment. PMID:26189585

  18. Expression of the Saccharomyces cerevisiae glycoprotein invertase in mouse fibroblasts: glycosylation, secretion, and enzymatic activity

    International Nuclear Information System (INIS)

    Oligosaccharide processing is controlled by host- and protein-dependent factors. To increase our understanding of the relative contribution of those factors the authors studied the glycosylation of yeast invertase expressed in a heterologous system. Invertase synthesized in psi-2 cells (an NIH 3T3-derived packaging line) is secreted efficiently, enzymatically active, and heavily glycosylated. It was estimated that the protein contains 8 or 9 carbohydrate chains. Two classes can be observed, of an approximate size of 100-110 kDa and 115-130 kDa, respectively. The size differences are due to differences in glycosylation. The smaller class contains two high-mannose carbohydrate chains; the remainder is of the complex type, sialylated and most likely tri- or tetraantennary. This profile parallels the situation observed with invertase glycosylation in yeast, where 2 of 9 or 10 chains remain unprocessed. The larger size class of invertase expressed in mouse fibroblasts has a different profile, since it contains probably only complex-type glycans. There are no apparent differences, however, in the size of the protein backbone between the two size classes. When invertase is synthesized in the presence of the mannosidase inhibitor 1-deoxymannojirimycin, processing is blocked completely. The glucosidase inhibitor 1-deoxynojirimycin does not inhibit processing completely. The glycosylation inhibitor tunicamycin prevents secretion of invertase completely when cells are cultured at 370C. At 260C, however, nonglycosylated invertase can be detected in the medium. These data suggest that glycosylation of invertase seems to be essential for the early steps of the secretory pathway but is less critical for later events

  19. Enhanced enzymatic activity of glycerol-3-phosphate dehydrogenase from the cryophilic Saccharomyces kudriavzevii.

    Directory of Open Access Journals (Sweden)

    Bruno M Oliveira

    Full Text Available During the evolution of the different species classified within the Saccharomyces genus, each one has adapted to live in different environments. One of the most important parameters that have influenced the evolution of Saccharomyces species is the temperature. Here we have focused on the study of the ability of certain species as Saccharomyces kudriavzevii to grow at low temperatures, in contrast to Saccharomyces cerevisiae. We observed that S. kudriavzevii strains isolated from several regions are able to synthesize higher amounts of glycerol, a molecule that has been shown to accumulate in response to freeze and cold stress. To explain this observation at the molecular level we studied the expression of glycerol biosynthetic pathway genes and we observed a higher expression of GPD1 gene in S. kudriavzevii compared to S. cerevisiae in micro-vinification conditions. We observed higher enzymatic activity of Gpd1p in S. kudriavzevii in response to osmotic and cold stress. Also, we determined that S. kudriavzevii Gpd1p enzyme presents increased catalytic properties that will contribute to increase glycerol production. Finally, we evaluated the glycerol production with S. cerevisiae, S. kudriavzevii or a recombinant Gpd1p variant in the same background and observed that the S. kudriavzevii enzyme produced increased glycerol levels at 12 or 28°C. This suggests that glycerol is increased in S. kudriavzevii mainly due to increased V max of the Gpd1p enzyme. All these differences indicate that S. kudriavzevii has changed the metabolism to promote the branch of the glycolytic pathway involved in glycerol production to adapt to low temperature environments and maintain the NAD(+/NADH ratio in alcoholic fermentations. This knowledge is industrially relevant due to the potential use, for example, of S. cerevisiae-S. kudriavzevii hybrids in the wine industry where glycerol content is an important quality parameter.

  20. Effect of a short period of phosphate deprivation on anti-oxidative enzymatic activities in bean plants.

    OpenAIRE

    Russo, Marco A; Malusa', Eligio; Verde, Elisabetta; Iacona, Riccardo; Bellomia, Laura; Belligno, Adalgisa

    2009-01-01

    We subjected bean plants (Phaseolus vulgaris L. ‘Bianco di Bagnasco’) to phosphate deprivation stress and studied the enzymatic activity of some major anti-oxidative enzymes in both leaves and roots extracts. P deprivation provoked a significant decrease of catalase activity in leaves of 14-day-old plants, but no differences were measured after prolonged P deprivation. The enzyme activity in roots was not affected by the withdrawal of P from the culture medium. The deprivation of P from the g...

  1. Effect of commercial cellulases and refining on kraft pulp properties: correlations between treatment impacts and enzymatic activity components.

    Science.gov (United States)

    Cui, Li; Meddeb-Mouelhi, Fatma; Laframboise, François; Beauregard, Marc

    2015-01-22

    The importance of enzymes as biotechnological catalysts for paper industry is now recognized. In this study, five cellulase formulations were used for fibre modification. The number of PFI revolutions decreased by about 50% while achieving the same freeness value (decrease in CSF by 200 mL) with the enzymatic pretreatment. The physical properties of handsheets were modified after enzymatic pretreatment followed by PFI refining. A slight decrease in tear strength was observed with enzymes C1 and C4 at pH 7 while the most decrease in tear was observed after C2, C3, C5 treatments. C1 and C4 which had xylanase activity improved paper properties, while other enzymes had a negative impact. Therefore, the intricate balance between cellulolytic and hemicellulolytic activity is the key to optimizing biorefining and paper properties. It was also observed that C1 impact was pH dependent, which supports the importance of pH in developing an enzymatic strategy for refining energy reduction. PMID:25439885

  2. New eutectic ionic liquids for lipase activation and enzymatic preparation of biodiesel†

    Science.gov (United States)

    Zhao, Hua; Baker, Gary A.; Holmes, Shaletha

    2012-01-01

    The enzymatic preparation of biodiesel has been hampered by the lack of suitable solvents with desirable properties such as high lipase compatibility, low cost, low viscosity, high biodegradability, and ease of product separation. Recent interest in using ionic liquids (ILs) as advanced reaction media has led to fast reaction rates and high yields in the enzymatic synthesis of biodiesel. However, conventional (i.e., cation–anion paired) ILs based on imidazolium and other quaternary ammonium salts remain too expensive for wide application at industrial scales. In this study, we report on newly-synthesized eutectic ILs derived from choline acetate or choline chloride coupled with biocompatible hydrogen-bond donors, such as glycerol. These eutectic solvents have favorable properties including low viscosity, high biodegradability, and excellent compatibility with Novozym® 435, a commercial immobilized Candida antarctica lipase B. Furthermore, in a model biodiesel synthesis system, we demonstrate high reaction rates for the enzymatic transesterification of Miglyol® oil 812 with methanol, catalyzed by Novozym® 435 in choline acetate/glycerol (1 : 1.5 molar ratio). The high conversion (97%) of the triglyceride obtained within 3 h, under optimal conditions, suggests that these novel eutectic solvents warrant further exploration as potential media in the enzymatic production of biodiesel. PMID:21283901

  3. Effects of host plants on digestive enzymatic activities and some components involved in intermediary metabolism of Chrysodeixis chalcites (Lepidoptera: Noctuidae

    Directory of Open Access Journals (Sweden)

    M. Mardani-Talaee

    2014-12-01

    Full Text Available Chrysodeixis chalcites (Esper is a serious pest that causes devastating damages in infested areas to many fruits, vegetables, ornamental crops and weeds. In the current study, effects of three host plants including lemon balm (Melissa officinalis L.; corn (Zea mays L. and dill (Anethum graveolens L. were determined on digestive enzyme activities and intermediary metabolism of C. chalcites larvae. The highest activities of α-amylase, glucosidases and specific proteases were observed in the larvae fed on dill. Our results showed that C. chalcites larvae had the highest TAG-lipase activity on corn in comparison with other host plants. Significant differences were found among enzymatic activities of acid (ACP and alkaline phosphatases, aspartate aminotransferases and lactate dehydrogenase (LDH in the haemolymph of C. chalcites larvae reared on lemon balm, corn and dill, respectively, although activity of alanine aminotransferase showed no statistically significant differences among different host plants. The enzymatic activity of ACP significantly decreased on dill in comparison with lemon balm and corn. The activity of LDH significantly increased on dill compared with other host plants. These results revealed that dill (A. graveolens is the most appropriate host plant for larvae of C. chalcites as evidenced by the highest digestive enzyme activities and intermediary metabolism.

  4. Priming effects and enzymatic activity in Israeli soils under treated wastewater and freshwater irrigation

    Science.gov (United States)

    Anissimova, Marina; Heinze, Stefanie; Chen, Yona; Tarchitzky, Jorge; Marschner, Bernd

    2014-05-01

    Irrigation of soils with treated wastewater (TWW) directly influences microbial processes of soil. TWW contains easily decomposable organic material, which can stimulate the activity of soil microorganisms and, as a result, lead to the excessive consumption of soil organic carbon pool. We investigated the effects of irrigation with TWW relative to those of irrigation with freshwater (FW) on the microbial parameters in soils with low (7%) and medium (13%) clay content in a lysimeter experiment. The objectives of our study were to (i) determine the impact of water quality on soil respiration and enzymatic activity influenced by clay content and depth, and (ii) work out the changes in the turnover of soil organic matter (PE, priming effects). Samples were taken from three soil depths (0-10, 10-20, and 40-60 cm). Soil respiration and PE were determined in a 21-days incubation experiment after addition of uniformly 14C-labeled fructose. Activity of 10 extracellular enzymes (EEA, from C-, N-, P-, and S-cycle), phenol oxidase and peroxidase activity (PO+PE), and dehydrogenase activity (DHA) were assayed. Microbial Community-Level Physiological Profiles (CLPP) using four substrates, and microbial biomass were determined. The results showed that the clay content acted as the main determinative factor. In the soil with low clay content the water quality had a greater impact: the highest PE (56%) was observed in the upper layer (0-10cm) under FW irrigation; EEA of C-, P-, and S-cycles was significantly higher in the upper soil layer under TWW irrigation. Microbial biomass was higher in the soil under TWW irrigation and decreased with increasing of depth (50 μg/g soil in the upper layer, 15 μg/g soil in the lowest layer). This tendency was also observed for DHA. Contrary to the low clay content, in the soil with medium clay content both irrigation types caused the highest PE in the lowest layer (65% under FW irrigation, 48% under TWW irrigation); the higher substrate

  5. Novel roles for well-known players: from tobacco mosaic virus pests to enzymatically active assemblies.

    Science.gov (United States)

    Koch, Claudia; Eber, Fabian J; Azucena, Carlos; Förste, Alexander; Walheim, Stefan; Schimmel, Thomas; Bittner, Alexander M; Jeske, Holger; Gliemann, Hartmut; Eiben, Sabine; Geiger, Fania C; Wege, Christina

    2016-01-01

    monitoring blood sugar concentrations, might profit particularly from the presence of TMV rods: Their surfaces were recently shown to stabilize enzymatic activities upon repeated consecutive uses and over several weeks. This review gives the reader a ride through strikingly diverse achievements obtained with TMV-based particles, compares them to the progress with related viruses, and focuses on latest results revealing special advantages for enzyme-based biosensing formats, which might be of high interest for diagnostics employing 'systems-on-a-chip'. PMID:27335751

  6. Molecular Basis for Enzymatic Sulfite Oxidation -- HOW THREE CONSERVED ACTIVE SITE RESIDUES SHAPE ENZYME ACTIVITY

    Energy Technology Data Exchange (ETDEWEB)

    Bailey, Susan; Rapson, Trevor; Johnson-Winters, Kayunta; Astashkin, Andrei; Enemark, John; Kappler, Ulrike

    2008-11-10

    Sulfite dehydrogenases (SDHs) catalyze the oxidation and detoxification of sulfite to sulfate, a reaction critical to all forms of life. Sulfite-oxidizing enzymes contain three conserved active site amino acids (Arg-55, His-57, and Tyr-236) that are crucial for catalytic competency. Here we have studied the kinetic and structural effects of two novel and one previously reported substitution (R55M, H57A, Y236F) in these residues on SDH catalysis. Both Arg-55 and His-57 were found to have key roles in substrate binding. An R55M substitution increased Km(sulfite)(app) by 2-3 orders of magnitude, whereas His-57 was required for maintaining a high substrate affinity at low pH when the imidazole ring is fully protonated. This effect may be mediated by interactions of His-57 with Arg-55 that stabilize the position of the Arg-55 side chain or, alternatively, may reflect changes in the protonation state of sulfite. Unlike what is seen for SDHWT and SDHY236F, the catalytic turnover rates of SDHR55M and SDHH57A are relatively insensitive to pH (~;;60 and 200 s-1, respectively). On the structural level, striking kinetic effects appeared to correlate with disorder (in SDHH57A and SDHY236F) or absence of Arg-55 (SDHR55M), suggesting that Arg-55 and the hydrogen bonding interactions it engages in are crucial for substrate binding and catalysis. The structure of SDHR55M has sulfate bound at the active site, a fact that coincides with a significant increase in the inhibitory effect of sulfate in SDHR55M. Thus, Arg-55 also appears to be involved in enabling discrimination between the substrate and product in SDH.

  7. Enzymatic activity of fungi of the genus Candida isolated from the skin and the digestive tract in people and from municipal sewage

    OpenAIRE

    Maria Dynowska; Ewa Sucharzewska; Anna Biedunkiewicz

    2014-01-01

    Enzymatic activities of C. albicans, C. guilliermondii and C. tropicalis from the skin and the digestive tract and from municipal sewage were compared using the API ZYM system (bio Méricux). Aclivities were examined in relalion to potential pathogenicity of these fungi strictly connected with their enzymatic properties, especilly the production of proteolytic and lipolytic enzymes, as well as acidic and basic phosphatase. The proteolytic activity was high in strains of C. albicans from the di...

  8. Generation of recombinant, enzymatically active human thyroid peroxidase and its recognition by antibodies in the sera of patients with Hashimoto's thyroiditis.

    OpenAIRE

    Kaufman, K D; Rapoport, B.; Seto, P; Chazenbalk, G D; Magnusson, R P

    1989-01-01

    A full-length cDNA clone for human thyroid peroxidase (TPO) inserted into the mammalian cell expression vector pECE was stably transfected into Chinese hamster ovary (CHO) cells. Clones were assayed for human TPO mRNA, TPO protein, and TPO enzymatic activity. One subclone, expressing the highest TPO enzymatic activity, was used in further studies. FACS analysis of these cells preincubated in Hashimoto's serum revealed approximately 100-fold greater fluorescence compared with controls, indicat...

  9. Effects of different bulking agents on the maturity, enzymatic activity, and microbial community functional diversity of kitchen waste compost.

    Science.gov (United States)

    Wang, Xiaojuan; Zhang, Wenwei; Gu, Jie; Gao, Hua; Qin, Qingjun

    2016-10-01

    Aerobic composting is an effective method for the disposal and utilization of kitchen waste. However, the addition of a bulking agent is necessary during kitchen waste composting because of its high moisture content and low C/N ratio. In order to select a suitable bulking agent, we investigated the influence of leaf litter (LL), sawdust (SD), and wheat straw (WS) on the enzymatic activity, microbial community functional diversity, and maturity indices during the kitchen waste composting process. The results showed that the addition of WS yielded the highest maturity (the C/N ratio decreased from 25 to 13, T value = 0.5, and germination index (GI) = 114.7%), whereas the compost containing SD as a bulking agent had the lowest maturity (GI = 32.4%). The maximum cellulase and urease activities were observed with the WS treatment on day 8, whereas the SD treatment had the lowest cellulase activity and the LL treatment had the lowest urease activity. The compost temperature and microbial activity (as the average well color development) showed that bulking the composts with SD prolonged the composting process. The diversity index based on the community-level physiological profile showed that the composts bulked with LL and WS had greater microbial community functional diversity compared with those bulked with SD. Thus, the maturity indexes and enzymatic activities suggest that WS is a suitable bulking agent for use in kitchen waste composting systems. PMID:26895274

  10. Enzymatic synthesis of dimeric glycomimetic ligands of NK cell activation receptors

    Czech Academy of Sciences Publication Activity Database

    Drozdová, Anna; Bojarová, Pavla; Křenek, Karel; Weignerová, Lenka; Hensen, B.; Elling, L.; Christensen, H.; Jensen, H.H.; Pelantová, Helena; Kuzma, Marek; Bezouška, Karel; Krupová, Monika; Adámek, David; Slámová, Kristýna; Křen, Vladimír

    2011-01-01

    Roč. 346, č. 12 (2011), s. 1599-1609. ISSN 0008-6215 R&D Projects: GA ČR GP203/09/P024; GA ČR GD305/09/H008; GA ČR GA303/09/0477 Institutional research plan: CEZ:AV0Z50200510 Keywords : beta-N-Acetylhexosaminidase * alactosyltransferase * Enzymatic glycosylation Subject RIV: CE - Biochemistry Impact factor: 2.332, year: 2011

  11. Predictive modelling of growth and measurement of enzymatic synthesis and activity by a cocktail of selected Enterobacteriaceae and Aeromonas hydrophila.

    Science.gov (United States)

    Braun, P; Sutherland, J P

    2005-11-25

    The possibility was examined of developing a predictive model that would predict food spoilage by combining microbial growth (increase in cellular number) with extracellular enzymatic activity of a cocktail of five strains of Enterobacteriaceae: Escherichia coli, Enterobacter agglomerans, Klebsiella oxytoca, Klebsiella pneumoniae and Proteus vulgaris and one Aeromonas hydrophila strain. Estimations of growth and enzyme activity were made within a three-dimensional matrix of conditions: temperature 2-20 degrees C, pH value 4.0-7.5 and water activity (a(w)) 0.95-0.995. A mathematical model was constructed which predicted growth based on increases in cell number. However, although notable effects of extracellular lipases and proteases were detected, it was not possible to model enzymatic activity and prepare a combined model because the data did not follow the characteristic profile that would allow curve-fitting. Nevertheless, the model for microbial growth and information relating to enzyme activity will be made freely available in a database on the internet. PMID:16154655

  12. Changes in Enzymatic Activity and Quality Attributes of Late-mature Peaches in Response to Controlled Atmosphere Conditions

    Institute of Scientific and Technical Information of China (English)

    TIAN Shi-ping; XU Yong; JIANG Ai-li; WANG Yi

    2002-01-01

    Late-mature peaches (Prunus persica L. Batsch, cv. Dongxuemi ) were stored in air,modification atmosphere (MA) and controlled atmospheres (CA) at 1℃ to investigate enzymatic activities,flesh browning, quality attributes and fruit storability during storage periods. Peaches stored in CA conditions showed significant lower activities of polyphenol oxidase (PPO) and peroxidase (POD) compared to that kept in MA and in air during the early period of storage. CA treatments indicated a better result in maintaining fruit firmness, color and titratable acidity than MA and control treatments. But there was no significant difference in vitamin C and total soluble solids of peaches stored in CA, MA and air. Increasing PPO activity obviously stimulated flesh browning. Peach fruits stored in CA conditions showed a lower activity of POD and higher firmness in the early period of storage. The peaches stored in 5%CO2 + 5%O2 and 10%CO2 + 5%O2 for 92 days had a good quality without any off-flavor. The results indicated CA conditions obviously inhibited the enzymatic browning, delayed senescence and maintained quality of late-mature peaches.

  13. Inhibitory effect of pinostrobin from Renealmia alpinia, on the enzymatic and biological activities of a PLA2.

    Science.gov (United States)

    Gómez-Betancur, Isabel; Pereañez, Jaime Andrés; Patiño, Arley Camilo; Benjumea, Dora

    2016-08-01

    Pinostrobin is a flavanone isolated from Renealmia alpinia, a plant used in folk medicine to treat snakebites. We tested the inhibitory ability of pinostrobin on the enzymatic, anticoagulant, myotoxic and edema-inducing activities of a PLA2 isolated from Crotalus durissus cumanensis venom. The compound displayed IC50 values of 1.76mM and 1.85mM (95% Confidence intervals: 1.34-2.18 and 1.21-2.45) on the PLA2 enzymatic activity, when either aggregated or monodispersed substrates were used, respectively. When mice were injected with PLA2 preincubated with 0.4, 2.0 and 4.0mM of pinostrobin, myotoxic activity induced by the PLA2 was inhibited up to 87%. Nevertheless, these values decreased up to 56% when the pinostrobin was injected into muscle after PLA2. Pinostrobin inhibited edema-forming and anticoagulant activities of the PLA2. In order to have insights on the mode of action of pinostrobin, intrinsic fluorescence and ultraviolet studies were performed. Results suggest that pinostrobin interacts directly with the PLA2. These findings were supported by molecular docking results, which suggested that pinostrobin forms hydrogen bonds with residues His48 and Asp49 of PLA2, besides, a π-π stacking interactions with those of residues Phe5 and Trp31, and rings C of flavanone and Tyr52 of the toxin. PMID:27109758

  14. Contrasting effects of untreated textile wastewater onto the soil available nitrogen-phosphorus and enzymatic activities in aridisol.

    Science.gov (United States)

    Arif, Muhammad Saleem; Riaz, Muhammad; Shahzad, Sher Muhammad; Yasmeen, Tahira; Buttler, Alexandre; Garcıa-Gil, Juan Carlos; Roohi, Mahnaz; Rasool, Akhtar

    2016-02-01

    Water shortage and soil qualitative degradation are significant environmental problems in arid and semi-arid regions of the world. The increasing demand for water in agriculture and industry has resulted in the emergence of wastewater use as an alternative in these areas. Textile wastewater is produced in surplus amounts which poses threat to the environment as well as associated flora and fauna. A 60-day incubation study was performed to assess the effects of untreated textile wastewater at 0, 25, 50, 75, and 100% dilution levels on the physico-chemical and some microbial and enzymatic properties of an aridisol soil. The addition of textile wastewater provoked a significant change in soil pH and electrical conductivity and soil dehydrogenase and urease activities compared to the distilled-water treated control soil. Moreover, compared to the control treatment, soil phosphomonoesterase activity was significantly increased from 25 to 75% application rates, but decreased at 100% textile wastewater application rate. Total and available soil N contents increased significantly in response to application of textile wastewater. Despite significant increases in the soil total P contents after the addition of textile wastewater, soil available P content decreased with increasing concentration of wastewater. Changes in soil nutrient contents and related enzymatic activities suggested a dynamic match between substrate availability and soil N and P contents. Aridisols have high fixation and low P availability, application of textile wastewater to such soils should be considered only after careful assessment. PMID:26787271

  15. Evaluation of oil removal efficiency and enzymatic activity in some fungal strains for bioremediation of petroleum-polluted soils

    Directory of Open Access Journals (Sweden)

    Mohsenzadeh Fariba

    2012-12-01

    Full Text Available Abstract Background Petroleum pollution is a global disaster and there are several soil cleaning methods including bioremediation. Methods In a field study, fugal strains were isolated from oil-contaminated sites of Arak refinery (Iran and their growth ability was checked in potato dextrose agar (PDA media containing 0-10% v/v crude oil, the activity of three enzymes (Catalase, Peroxidase and Phenol Oxidase was evaluated in the fungal colonies and bioremediation ability of the fungi was checked in the experimental pots containing 3 kg sterilized soil and different concentrations of petroleum (0-10% w/w. Results Four fungal strains, Acromonium sp., Alternaria sp., Aspergillus terreus and Penicillium sp., were selected as the most resistant ones. They were able to growth in the subjected concentrations and Alternaria sp. showed the highest growth ability in the petroleum containing media. The enzyme assay showed that the enzymatic activity was increased in the oil-contaminated media. Bioremediation results showed that the studied fungi were able to decrease petroleum pollution. The highest petroleum removing efficiency of Aspergillus terreus, Penicillium sp., Alternaria sp. and Acromonium sp. was evaluated in the 10%, 8%, 8% and 2% petroleum pollution respectively. Conclusions Fungi are important microorganisms in decreasing of petroleum pollution. They have bioremediation potency that is related to their enzymatic activities.

  16. Evaluation of Oil Removal Efficiency and Enzymatic Activity in Some fungal Strains for Bioremediation of Petroleum-Polluted Soils

    Directory of Open Access Journals (Sweden)

    Fariba Mohsenzadeh

    2012-12-01

    Full Text Available Background: Petroleum pollution is a global disaster and there are several soil cleaning methods including bioremediation.Methods: In a field study, fugal strains were isolated from oil-contaminated sites of Arak refinery (Iran and their growth ability was checked in potato dextrose agar (PDA media containing 0-10% v/v crude oil, the activity of three enzymes (Catalase, Peroxidase and Phenol Oxidase was evaluated in the fungal colonies and bioremediation ability of the fungi was checked in the experimental pots containing 3 kg sterilized soil and different concentrations of petroleum (0-10% w/w.Results: Four fungal strains, Acromonium sp., Alternaria sp., Aspergillus terreus and Penicillium sp., were selected asthe most resistant ones. They were able to growth in the subjected concentrations and Alternaria sp. showed thehighest growth ability in the petroleum containing media. The enzyme assay showed that the enzymatic activity was increased in the oil-contaminated media. Bioremediation results showed that the studied fungi were able to decrease petroleum pollution. The highest petroleum removing efficiency of Aspergillus terreus, Penicillium sp.,Alternaria sp. and Acromonium sp. was evaluated in the 10%, 8%, 8% and 2% petroleum pollution respectively.Conclusions: Fungi are important microorganisms in decreasing of petroleum pollution. They have bioremediation potency that is related to their enzymatic activities.

  17. Enzymatic degradation of aromatic hydrocarbon intermediates using a recombinant dioxygenase immobilized onto surfactant-activated carbon nanotube.

    Science.gov (United States)

    Suma, Yanasinee; Lim, Heejun; Kwean, Oh Sung; Cho, Suyeon; Yang, Junwon; Kim, Yohan; Kang, Christina S; Kim, Han S

    2016-06-01

    This study examined the enzymatic decomposition of aromatic hydrocarbon intermediates (catechol, 4-chlorocatechol, and 3-methylcatechol) using a dioxygenase immobilized onto single-walled carbon nanotube (SWCNT). The surfaces of SWCNTs were activated with surfactants. The dioxygenase was obtained by recombinant technique: the corresponding gene was cloned from Arthrobacter chlorophenolicus A6, and the enzyme was overexpressed and purified subsequently. The enzyme immobilization yield was 62%, and the high level of enzyme activity was preserved (60-79%) after enzyme immobilization. Kinetic analyses showed that the substrate utilization rates and the catalytic efficiencies of the immobilized enzyme for all substrates (target aromatic hydrocarbon intermediates) tested were similar to those of the free enzyme, indicating that the loss of enzyme activity was minimal during enzyme immobilization. The immobilized enzyme was more stable than the free enzyme against abrupt changes in pH, temperature, and ionic strength. Moreover, it retained high enzyme activity even after repetitive use. PMID:26810145

  18. An antiapoptotic role for telomerase RNA in human immune cells independent of telomere integrity or telomerase enzymatic activity

    OpenAIRE

    Gazzaniga, Francesca S.; Elizabeth H. Blackburn

    2014-01-01

    Telomerase RNA component hTR, but not the core enzymatic protein component hTERT, protects T cells from apoptosis.hTR prevents dexamethasone-induced apoptosis specifically when in a telomerase enzymatically inactive state.

  19. IMPACT OF ALTITUDES ON SOIL CHARACTERISTICS AND ENZYMATIC ACTIVITIES IN FOREST AND FALLOW LANDS OF ALMORA DISTRICT OF CENTRAL HIMALAYA

    Directory of Open Access Journals (Sweden)

    B. R. Maurya

    2014-03-01

    Full Text Available Abstract: Altitude is one of the major topographical factors which influence the fertility status of soil. Population explosion has rooted deforestation at different altitudes to bring more area under cultivation leading to fallow lands. Objective of this study was to assess the impact of altitude on electro-chemical properties and enzymatic activities of forest and fallow land soils of Almora district of Central Himalaya. Seventy soil samples were collected from different altitudes of forest and fallow lands of Almora. Electro-chemical properties, soil microbial biomass carbon (SMBC, β-glucosidase, dehydrogenase and phosphatase activities were determined following the standard procedures. Both forest and fallow land soils were acidic in nature. Content of organic carbon, nitrogen, phosphorous and potassium were high in range and increased with altitudes. Average content of total nitrogen, phosphorous and potassium in forest lands was 2115.39, 111.89 and 2189.36 kg ha 1, respectively. Similarly for the fallow lands the corresponding values were 1491.27, 80.26 and 2650.75 kg ha-1, respectively. SMBC of forest soil was positively and significantly correlated with β-glucosidase, dehydrogenase and phosphatase activities but in fallow land soil, it was significantly correlated only with dehydrogenase activities (r=0.69. All enzymes of forest soil were positively and significantly correlated with organic carbon, total nitrogen and total potassium. For fallow lands dehydrogenase activity was positively and significantly correlated with nitrogen (r=0.65 and potassium (r=0.76 while phosphatase activity was negatively and significantly correlated with total phosphorous (r= -0.71. Concluded that altitude directly or indirectly has influenced fertility status, SMBC and enzymatic activities of forest as well fallow land soils.

  20. Modulated expression and enzymatic activities of Darkbarbel catfish, Pelteobagrus vachelli for oxidative stress induced by acute hypoxia and reoxygenation.

    Science.gov (United States)

    Zhang, Guosong; Mao, Jianqiang; Liang, Fenfei; Chen, Jiawei; Zhao, Cheng; Yin, Shaowu; Wang, Li; Tang, Zhonglin; Chen, Shuqiao

    2016-05-01

    Large changes in oxygen availability in aquatic environments, ranging from anoxia through to hyperoxia, can lead to corresponding wide variation in the production of reactive oxygen species (ROS) by fish with aquatic respiration. In order to evaluate the effects of hypoxia and reoxygenation on oxidative stress in fish, the mRNA and protein expression of SODs (Cu/Zn-SOD and Mn-SOD) as well as indices (CP, LPO and MDA) and enzymatic activities (SOD, CAT, GPx, GR and GST) were analyzed in liver and brain tissues of Pelteobagrus vachelli. Predominant expression of PvSOD2 was detected in heart, brain, and liver. In contrast, PvSOD1 was highly expressed in liver. Based on the expression patterns of above parameters, we inferred that brain tissue of P. vachelli under 0.7 mg/L degree of acute hypoxia condition could experience hypometabolic states or no suffering stress, but brain tissue has effective mechanisms to minimize or prevent oxidative stress during the transition from hypoxia to reoxygenation. Our results also demonstrated an increased expression of SODs and enzymatic activities for oxidative stress in liver under hypoxic conditions, which supports the hypothesis that anticipatory preparation takes place in order to deal with the encountered oxidative stress during the recovery from hypoxia as proposed by M. Hermes-Lima. Therefore, this study will provide a clue to better understand the action mode of antioxidant genes and enzymes under oxidative stress in fish. PMID:26945243

  1. Mitochondrial intermediate peptidase: Expression in Escherichia coli and improvement of its enzymatic activity detection with FRET substrates

    International Nuclear Information System (INIS)

    In the present study, soluble, functionally-active, recombinant human mitochondrial intermediate peptidase (hMIP), a mitochondrial metalloendoprotease, was expressed in a prokaryotic system. The hMIP fusion protein, with a poly-His-tag (6x His), was obtained by cloning the coding region of hMIP cDNA into the pET-28a expression vector, which was then used to transform Escherichia coli BL21 (DE3) pLysS. After isolation and purification of the fusion protein by affinity chromatography using Ni-Sepharose resin, the protein was purified further using ion exchange chromatography with a Hi-trap resource Q column. The recombinant hMIP was characterized by Western blotting using three distinct antibodies, circular dichroism, and enzymatic assays that used the first FRET substrates developed for MIP and a series of protease inhibitors. The successful expression of enzymatically-active hMIP in addition to the FRET substrates will contribute greatly to the determination of substrate specificity of this protease and to the development of specific inhibitors that are essential for a better understanding of the role of this protease in mitochondrial functioning.

  2. Human SOD2 modification by dopamine quinones affects enzymatic activity by promoting its aggregation: possible implications for Parkinson's disease.

    Directory of Open Access Journals (Sweden)

    Elisa Belluzzi

    Full Text Available Mitochondrial dysfunction and oxidative stress are considered central in dopaminergic neurodegeneration in Parkinson's disease (PD. Oxidative stress occurs when the endogenous antioxidant systems are overcome by the generation of reactive oxygen species (ROS. A plausible source of oxidative stress, which could account for the selective degeneration of dopaminergic neurons, is the redox chemistry of dopamine (DA and leads to the formation of ROS and reactive dopamine-quinones (DAQs. Superoxide dismutase 2 (SOD2 is a mitochondrial enzyme that converts superoxide radicals to molecular oxygen and hydrogen peroxide, providing a first line of defense against ROS. We investigated the possible interplay between DA and SOD2 in the pathogenesis of PD using enzymatic essays, site-specific mutagenesis, and optical and high-field-cw-EPR spectroscopies. Using radioactive DA, we demonstrated that SOD2 is a target of DAQs. Exposure to micromolar DAQ concentrations induces a loss of up to 50% of SOD2 enzymatic activity in a dose-dependent manner, which is correlated to the concomitant formation of protein aggregates, while the coordination geometry of the active site appears unaffected by DAQ modifications. Our findings support a model in which DAQ-mediated SOD2 inactivation increases mitochondrial ROS production, suggesting a link between oxidative stress and mitochondrial dysfunction.

  3. Detection of endopeptidase activity and analysis of cleavage specificity using a radiometric solid-phase enzymatic assay

    International Nuclear Information System (INIS)

    A radiometric procedure to detect the presence of proteolytic enzymes and analyze their substrate specificity is described. The enzymatic activity is first measured by the release into solution of a radiolabeled reporter group from an immobilized peptidyl substrate. Two peptidyl substrates encompassing multiple cleavage sites, a derivative of Leu-enkephalin and a peptide related to the bait region of human α 2-macroglobulin, are prepared and linked via a spacer molecule to an insoluble support. The labeled peptides released are then separated by high-performance liquid chromatography. The position of the released peptides upon chromatography allows direct identification of the sites of cleavage. The assay, using a radioactive iodinated tyrosine residue as reporter group, is extremely sensitive (less than 0.02 pg/ml of trypsin), reproducible, and easy to perform while yielding unambiguous identification of the sites of cleavage. This assay can be used to detect the presence of enzymatic activities and/or of enzyme inhibitors. Furthermore, it can be easily adapted to detect from a variety of sources all four classes of enzymes known by using appropriate peptidyl substrate sequences, buffer, pH, and incubation conditions

  4. Oxidant and enzymatic antioxidant status (gene expression and activity) in the brain of chickens with cold-induced pulmonary hypertension

    Science.gov (United States)

    Hassanpour, Hossein; Khalaji-Pirbalouty, Valiallah; Nasiri, Leila; Mohebbi, Abdonnaser; Bahadoran, Shahab

    2015-11-01

    To evaluate oxidant and antioxidant status of the brain (hindbrain, midbrain, and forebrain) in chickens with cold-induced pulmonary hypertension, the measurements of lipid peroxidation, protein oxidation, antioxidant capacity, enzymatic activity, and gene expression (for catalase, glutathione peroxidase, and superoxide dismutases) were done. There were high lipid peroxidation/protein oxidation and low antioxidant capacity in the hindbrain of cold-induced pulmonary hypertensive chickens compared to control ( P hypertensive chickens, superoxide dismutase activity was decreased (forebrain, midbrain, and hindbrain), while catalase activity was increased (forebrain and midbrain) ( P hypertensive chickens. Probably, these situations in the oxidant and antioxidant status of the brain especially hindbrain may change its function at cardiovascular center and sympathetic nervous system to exacerbate pulmonary hypertension.

  5. Antioxidant activities and functional properties of enzymatic protein hydrolysates from defatted Camellia oleifera seed cake.

    Science.gov (United States)

    Li, Xu; Deng, Junlin; Shen, Shian; Li, Tian; Yuan, Ming; Yang, Ruiwu; Ding, Chunbang

    2015-09-01

    Seed cake protein (SCP) from Camellia oleifera was hydrolyzed by five commercial proteases (Flavorzyme, Trypsin, Neutrase, Papain, Alcalase). Amino acid composition, molecular weight distribution, antioxidant activity and functional property of the seed cake protein hydrolysates (SCPH) were investigated. Enzymatic hydrolysis improved protein solubility significantly but impaired the foaming and emulsifying property. Hydrolysate generated by alcalase had the highest hydrolysis degree (DH) and antioxidant activity, and displayed excellent protein solubility over wide range of pH, while hydrolysate prepared by flavorzyme showed better copper chelating capacity and emulsifying stability with low molecular weight distribution. Trypsin-treated SCPH showed better foaming property than original protein. The results indicated that enzyme type greatly influenced the molecular weight, functional property and antioxidant activity of SCPH. It was also found that electing appropriate protease and controlling the DH could be enhanced or reduced functional property according to actual applications. PMID:26344981

  6. Bioconversion and enzymatic activities of neurospora sitophila grown under solid state and submerged fermentation on Sago Hamps

    International Nuclear Information System (INIS)

    N.Sitophila was grown under controlled conditions of solid state and submerged fermentation on Sago Hampas. The optimum conditions of protein enrichment previously established for sugar beet pulp was used for this study. Under this condition the protein content of Sago Hampas under solid state increased from 1.4 to 14.45% (W/W) whereas for Sago Hampas and Sago starch, the protein content under submerged condition increased from 1.4% (W/W) and 0.7% (W/W) to 18.56% (W/W) and 43/16% (W/W) based on dry weight of product respectively. The cellulase, a-amylase and glucoamylase activities of N.Sitophila under solid state condition on Sago Hampas were, 9.0, 0.6 and 11.8 U/g of wet fermented solid respectively. the enzymatic activities were also measured under submerged fermentation using both Sago Hampas and Sago starch as substrate

  7. A flexible loop controlling the enzymatic activity and specificity in a glycosyl hydrolase family 19 endochitinase from barley seeds

    DEFF Research Database (Denmark)

    Fukamizo, Tamo; Miyake, Ryoh; Tamura, Atsushi;

    2009-01-01

    To examine the role of the loop structure consisting of residues 70-82 (70-82 loop) localized to + 3/4 subsite of the substrate binding cleft of a family GH-19 endochitinase from barley seeds, Trp72 and Trp82 were mutated, and the mutated enzymes (W72A, W82A, and W72A/W82A) were characterized....... Thermal stability and specific activities toward glycol chitin and chitin hexasaccharide were significantly affected by the individual mutations. When N-acetylglucosamine hexamer was hydrolyzed by the wild type, the ß-anomer of the substrate was preferentially hydrolyzed, producing the trimer...... predominantly and the dimer and tetramer in lesser amounts. When the mutated enzymes were used instead of the wild type, the enzyme cleavage sites in the hexamer substrate were clearly shifted, and the ß-anomer selectivity was eliminated. The mutation effects on the enzymatic activity and stability were much...

  8. Evaluating the pollution from Mureş River on Arad-Pecica sector based on enzymatic activities from sediments (Western Romania

    Directory of Open Access Journals (Sweden)

    Sebastian TREITLI

    2011-05-01

    Full Text Available Seven sediment samples were collected from the river Mureş from the Arad-Pecica sector and were measured the following enzymatic activities: catalase, actual and potential dehydrogenase, urease and reduction of trivalent iron (Fe3+. All this activities were detected in all analyzed samples. Based on relative values of the enzymatic activities, the enzymatic indicator of the sediment quality (EISQ was also calculated. The lowest value of EISQ was observed in point P6 in close proximity to the discharge area of waste water resulted from water treatment plant of the Pecica town, which could indicate a heavy pollution from the water treatment plant of the town. The pollution of Mureş river on the analyzed sector can be determined by livestock farms, gravels and water treatment plants which do not work properly.

  9. Lactogenic Activity of an Enzymatic Hydrolysate from Octopus vulgaris and Carica papaya in SD Rats.

    Science.gov (United States)

    Cai, Bingna; Chen, Hua; Sun, Han; Sun, Huili; Wan, Peng; Chen, Deke; Pan, Jianyu

    2015-11-01

    The traditional Chinese medicine theory believes that octopus papaya soup can stimulate milk production in lactating women. The objective of this study was to determine whether dietary supplementation with an enzymatic hydrolysate of Octopus vulgaris and Carica papaya (EHOC) could increase milk production and nutritional indexes in Sprague Dawley (SD) rats. Female SD rats (n = 24) were fed a control diet (n = 8), EHOC-supplemented diet, or a positive control diet (Shengruzhi) from day 10 of pregnancy to day 10 of lactation. Maternal serum, mammary gland (day 10 of lactation), milk, and pup weight (daily) were collected for analysis. Results showed that the EHOC diet obviously elevated daily milk yield and pup weight compared to the control group (P supplemented dams were higher than those of the control group, especially the cholesterol, glucose, and IgG were higher by 44.98% (P < .001), 42.76% (P < .01), and 42.23% (P < .01), respectively. In conclusion, this article demonstrates that EHOC administration has beneficial effects on milk production in the dams and on performance of the dam and pup. These results indicate that EHOC could be explored as a potentially lactogenic nutriment for lactating women. PMID:26270883

  10. Protein-Rich Fraction of Cnidoscolus urens (L. Arthur Leaves: Enzymatic Characterization and Procoagulant and Fibrinogenolytic Activities

    Directory of Open Access Journals (Sweden)

    Yamara A. S. de Menezes

    2014-03-01

    Full Text Available Proteolytic enzymes are important macromolecules in the regulation of biochemical processes in living organisms. Additionally, these versatile biomolecules have numerous applications in the industrial segment. In this study we have characterized a protein-rich fraction of Cnidoscolus urens (L. Arthur leaves, rich in proteolytic enzymes, and evaluated its effects on the coagulation cascade. Three protein-rich fractions were obtained from the crude extract of C. urens leaves by precipitation with acetone. Fraction F1.0 showed higher proteolytic activity upon azocasein, and thus, was chosen for subsequent tests. The proteolytic activity of F1.0 on fibrinogen was dose-dependent and time-dependent. The extract demonstrated procoagulant activity on citrated plasma and reduced the APTT, not exerting effects on PT. Despite the fibrin(ogenolytic activity, F1.0 showed no defibrinogenating activity in vivo. The fraction F1.0 did not express hemorrhagic nor hemolytic activities. The proteolytic activity was inhibited by E-64, EDTA and in the presence of metal ions, and increased when pretreated with reducing agents, suggesting that the observed activity was mostly due to cysteine proteases. Several bands with proteolytic activity were detected by zymography with gelatin, albumin and fibrinogen. The optimal enzymatic activity was observed in temperature of 60 °C and pH 5.0, demonstrating the presence of acidic proteases. In conclusion, these results could provide basis for the pharmacological application of C. urens proteases as a new source of bioactive molecules to treat bleeding and thrombotic disorders.

  11. Enzymatic Hydrolysis of Lignocelluloses

    DEFF Research Database (Denmark)

    Kolasa, Marta; Ahring, Birgitte Kiær; Lübeck, Peter Stephensen;

    2010-01-01

    bonds. Cellulose can be degraded to simple sugar components by means of enzymatic hydrolysis. However, due to its complex, crystalline structure it is difficult to break it down and the cooperative action of a variety of cellulolytic enzymes is necessary. Fungi are known to have potential in production...... of a variety of cellulolytic enzymes. The aim of this work is to discover new thermostable and robust cellulolytic enzymes for improved enzymatic hydrolysis of biomass. For this purpose two screening methods are applied in different fungal strains with high cellulolytic activities: an expression...

  12. Site-Specific Bioconjugation of an Organometallic Electron Mediator to an Enzyme with Retained Photocatalytic Cofactor Regenerating Capacity and Enzymatic Activity

    Directory of Open Access Journals (Sweden)

    Sung In Lim

    2015-04-01

    Full Text Available Photosynthesis consists of a series of reactions catalyzed by redox enzymes to synthesize carbohydrates using solar energy. In order to take the advantage of solar energy, many researchers have investigated artificial photosynthesis systems mimicking the natural photosynthetic enzymatic redox reactions. These redox reactions usually require cofactors, which due to their high cost become a key issue when constructing an artificial photosynthesis system. Combining a photosensitizer and an Rh-based electron mediator (RhM has been shown to photocatalytically regenerate cofactors. However, maintaining the high concentration of cofactors available for efficient enzymatic reactions requires a high concentration of the expensive RhM; making this process cost prohibitive. We hypothesized that conjugation of an electron mediator to a redox enzyme will reduce the amount of electron mediators necessary for efficient enzymatic reactions. This is due to photocatalytically regenerated NAD(PH being readily available to a redox enzyme, when the local NAD(PH concentration near the enzyme becomes higher. However, conventional random conjugation of RhM to a redox enzyme will likely lead to a substantial loss of cofactor regenerating capacity and enzymatic activity. In order to avoid this issue, we investigated whether bioconjugation of RhM to a permissive site of a redox enzyme retains cofactor regenerating capacity and enzymatic activity. As a model system, a RhM was conjugated to a redox enzyme, formate dehydrogenase obtained from Thiobacillus sp. KNK65MA (TsFDH. A RhM-containing azide group was site-specifically conjugated to p-azidophenylalanine introduced to a permissive site of TsFDH via a bioorthogonal strain-promoted azide-alkyne cycloaddition and an appropriate linker. The TsFDH-RhM conjugate exhibited retained cofactor regenerating capacity and enzymatic activity.

  13. Dynamics of microbiological parameters, enzymatic activities and worm biomass production during vermicomposting of effluent treatment plant sludge of bakery industry.

    Science.gov (United States)

    Yadav, Anoop; Suthar, S; Garg, V K

    2015-10-01

    This paper reports the changes in microbial parameters and enzymatic activities during vermicomposting of effluent treatment plant sludge (ETPS) of bakery industry spiked with cow dung (CD) by Eisenia fetida. Six vermibins containing different ratios of ETPS and CD were maintained under controlled laboratory conditions for 15 weeks. Total bacterial and total fungal count increased upto 7th week and declined afterward in all the bins. Maximum bacterial and fungal count was 31.6 CFU × 10(6) g(-1) and 31 CFU × 10(4) g(-1) in 7th week. Maximum dehydrogenase activity was 1921 μg TPF g(-1) h(-1) in 9th week in 100 % CD containing vermibin, whereas maximum urease activity was 1208 μg NH4 (-)N g(-1) h(-1) in 3rd week in 100 % CD containing vermibin. The enzyme activity and microbial counts were lesser in ETPS containing vermibins than control (100 % CD). The growth and fecundity of the worms in different vermibins were also investigated. The results showed that initially biomass and fecundity of the worms increased but decreased at the later stages due to non-availability of the palatable feed. This showed that quality and palatability of food directly affect biological parameters of the system. PMID:25982984

  14. Longitudinal changes in PON1 enzymatic activities in Mexican-American mothers and children with different genotypes and haplotypes

    International Nuclear Information System (INIS)

    The paraoxonase 1 (PON1) enzyme prevents low-density lipoprotein oxidation and also detoxifies the oxon derivatives of certain neurotoxic organophosphate (OP) pesticides. PON1 activity in infants is low compared to adults, rendering them with lower metabolic and antioxidant capacities. We made a longitudinal comparison of the role of genetic variability on control of PON1 phenotypes in Mexican-American mothers and their children at the time of delivery (n = 388 and 338, respectively) and again 7 years later (n = 280 and 281, respectively) using generalized estimating equations models. At age 7, children's mean PON1 activities were still lower than those of mothers. This difference was larger in children with genotypes associated with low PON1 activities (PON1-108TT, PON1192QQ, and PON1-909CC). In mothers, PON1 activities were elevated at delivery and during pregnancy compared to 7 years later when they were not pregnant (p < 0.001). In non-pregnant mothers, PON1 polymorphisms and haplotypes accounted for almost 2-fold more variation of arylesterase (AREase) and chlorpyrifos-oxonase (CPOase) activity than in mothers at delivery. In both mothers and children, the five PON1 polymorphisms (192, 55, -108, -909, -162) explained a noticeably larger proportion of variance of paraoxonase activity (62-78%) than AREase activity (12.3-26.6%). Genetic control of PON1 enzymatic activity varies in children compared to adults and is also affected by pregnancy status. In addition to known PON1 polymorphisms, unidentified environmental, genetic, or epigenetic factors may also influence variability of PON1 expression and therefore susceptibility to OPs and oxidative stress.

  15. Antimicrobial effect and enzymatic activity of extract of Zingiber officinale Roscoe and stability in topical preparations

    OpenAIRE

    Gisele Mara Silva Gonçalves; Gustavo Henrique da Silva; Pedro Paulo Barros; Silvana Mariana Srebernich; Fernanda Rocha Tonon; Daniella Souza Fiore

    2014-01-01

    The rhizomes of common ginger (Zingiber officinale Roscoe) contain substances with antimicrobial activity and proteolytic enzymes and thus may have various pharmaceutical applications. The aim of the present study was to prepare Zingiber officinale Roscoe rhizome extracts for pharmaceutical use, preserving the proteolytic enzyme activity and testing the antimicrobial activity, in order to develop topical formulations. Two extracts were obtained - aqueous and glycolic ...

  16. Extracellular enzymatic activities of cold-adapted bacteria from polar oceans and effect of temperature and salinity on cell growth

    Institute of Scientific and Technical Information of China (English)

    Zeng Yinxin; Yu Yong; Chen Bo; Li Huirong

    2004-01-01

    The potential of 324 bacteria isolated from different habitats in polar oceans to produce a variety of extracellular enzymatic activities at low temperature was investigated. By plate assay, lipase, protease, amylase, gelatinase, agarase, chitinase or cellulase were detected. Lipases were generally present by bacteria living in polar oceans. Protease-producing bacteria held the second highest proportion in culturable isolates. Strains producing amylase kept a relative stable proportion of around 30% in different polar marine habitats. All 50 Arctic sea-ice bacteria producing proteases were cold-adapted strains, however, only 20% were psychrophilic. 98% of them could grow at 3% NaCl, and 56% could grow without NaCl. On the other hand, 98% of these sea-ice bacteria produced extracellular proteases with optimum temperature at or higher than 35℃, well above the upper temperature limit of cell growth. Extracellular enzymes including amylase, agarase, cellulase and lipase released by bacteria from seawater or sediment in polar oceans, most expressed maximum activities between 25 and 35℃. Among extracellular enzymes released by bacterial strain BSw20308, protease expressed maximum activity at 40℃, higher than 35℃ of polysaccharide hydrolases and 25℃ of lipase.

  17. Effects of Multiglycoside of Tripterygium Wilfordii (GTW) on Spermatogenic Ceils and Enzymatic Activities in Male Rats

    Institute of Scientific and Technical Information of China (English)

    田健; 周孝瑚

    1994-01-01

    This study was undertaken to observe consecutively the morphology of testis and epididymis and the changes of enzymatic activities of AKP, ACP, 3β-HSD and LDH-C4 in male rats treated with GTW 30 to 80 days. In addition, male antifertility effect and its possible reversibility were also observed. The results showed that GTW is a potential testicular toxicant in the animals. It can cause damage of the sperm cells with close relation to the treated time and dosage, Sloughing of sperm cells and many multinucleated giant cells in the seminiferous tubules were found on day 40 and 50 after the treatment. The tubules were hyperatrophic and the sperm cells were almost absent at the end of the study(80 days), No obvious morphological alteration was observed in the epididymal epithelial tissue, But changes of quality and number df the spermatozoa in epididymides were found prior to those of testes. Reduced number and abnormal cauda sperm were observed 30 days after the treatment, The ACP activity of Sertoli cells increased slightly, whereas the activities of ACP and LDH-C4 decreased gradually as the treated time prolonged. The 3β-HSD of Leydig cells was changed or subtle by m-phase or treatment and dramatically decreased at the end of treatment, The infertility caused by GTW was reversible 8 weeks after cessation of the treatment.

  18. Effects of Straw Processing Methods and Irrigation Sources on Enzymatic Activity of Soils under Winter Wheat

    Institute of Scientific and Technical Information of China (English)

    Zhiwei LU; Guofeng WAN; Zijun YANG; Lei HOU; Wenhui ZHANG

    2012-01-01

    [Objective] The aim was to study on effects of "straw returning and ir-re- turning" and "irrigation with ground water and water in the Yellow River" on changes of enzyme activity in soils under wheat at different developmental stages. [Method] Jimai 22, a kind of winter wheat, was made use of in fields to study on effects of " straw returning and Jr-returning" and "irrigation with ground water and water in the Yellow River" on changes of enzyme activity in soils under wheat in different devel- opmental stages. [Result] With advancement of developmental stage, urease activity of wheat in the four groups all showed the trend of "increasing-decreasing-increas- ing" and activities of invertase and phosphatase both changed from increasing to de- creasing. In addition, urease activities of soils in wheat roots were improved by straw returning in four developmental stages. Meanwhile, activity of soil enzyme was better promoted by irrigation with ground water than with water in the Yellow River, except in grain-filling stage. Before developmental stage, different processing meth- ods had a significant effect on phosphatase activity, for example, straw returning and ground water significantly enhanced activities of two kinds of phosphatase and pro- moted P absorption and transferring by plants and microorganisms in jointing stage; activity of acid phosphatase was higher in the group where irrigation with ground water and straw returning were adopted than those in the rest three groups in boot- ing stage. [Conclusion] The research laid a foundation for dynamic relationship among activity of soil enzyme, crop growth and microorganisms.

  19. Enzymatic activity of granulations tissues under low doses of radiation. Biochemical analysis in rats

    International Nuclear Information System (INIS)

    This paper was designed to investigate in the rat subcutaneous sponge-induced granulation tissue under low doses of X-ray, the activity of alkaline phosphatase, 5'nucleotide phosphodiesterase and adenosine triphosphatase (ATPase) enzymes. One hundred and fourteen Wistar rats were divided into three groups, as follows: Group I as control, Group II that received single 7,14 R in split-dosis immediately after sponge-implantation at the third and fifth days postoperatively. Biopsies were taken after 7, 11, 14, 21 and 28 days and the activity of the three enzymes was determined. The results have shown that in Group II alkaline phosphatase had higher activity in the 14th day of tissue evolution when compared to Groups I and III . The 5'nucleotide phosphodiesterase activity in Group I was similar in all days checked, although in Group II the enzyme showed higher activity in 7th day and lower in 21st. In Group III the activity was higher after 14 and 7 days and lower after 28 and 21 days. There was no observation of changing in adenosine triphosphatase (ATPase) activity when the three groups were compared. (author)

  20. Nanonets Derived from Turnip Mosaic Virus as Scaffolds for Increased Enzymatic Activity of Immobilized Candida antarctica Lipase B

    Science.gov (United States)

    Cuenca, Sol; Mansilla, Carmen; Aguado, Marta; Yuste-Calvo, Carmen; Sánchez, Flora; Sánchez-Montero, Jose M.; Ponz, Fernando

    2016-01-01

    Elongated flexuous plant viral nanoparticles (VNPs) represent an interesting platform for developing different applications in nanobiotechnology. In the case of potyviruses, the virion external surface is made up of helically arrayed domains of the viral structural coat protein (CP), repeated over 2000 times, in which the N- and C-terminal domains of each CP are projected toward the exterior of the external virion surface. These characteristics provide a chemical environment rich in functional groups susceptible to chemical conjugations. We have conjugated Candida antarctica lipase B (CALB) onto amino groups of the external surface of the potyvirus turnip mosaic virus (TuMV) using glutaraldehyde as a conjugating agent. Using this approach, TuMV virions were transformed into scaffolds for CALB nanoimmobilization. Analysis of the resulting structures revealed the formation of TuMV nanonets onto which large CALB aggregates were deposited. The functional enzymatic characterization of the CALB-bearing TuMV nanonets showed that CALB continued to be active in the nanoimmobilized form, even gaining an increased relative specific activity, as compared to the non-immobilized form. These novel virus-based nanostructures may provide a useful new approach to enzyme nanoimmobilization susceptible to be industrially exploited. PMID:27148295

  1. Identification of a novel enzymatic activity from lactic acid bacteria able to degrade biogenic amines in wine.

    Science.gov (United States)

    Callejón, S; Sendra, R; Ferrer, S; Pardo, I

    2014-01-01

    The main objectives of this study were the search for enzymatic activities responsible for biogenic amine (BA) degradation in lactic acid bacteria (LAB) strains isolated from wine, their identification, and the evaluation of their applicability for reducing BAs in wine. Fifty-three percent of the 76 LAB cell extracts showed activity against a mixture of histamine, tyramine, and putrescine when analyzed in-gel. The quantification of the degrading ability for each individual amine was tested in a synthetic medium and wine. Most of the bacteria analyzed were able to degrade the three amines in both conditions. The highest percentages of degradation in wine were those of putrescine: up to 41% diminution in 1 week. Enzymes responsible for amine degradation were isolated and purified from Lactobacillus plantarum J16 and Pediococcus acidilactici CECT 5930 strains and were identified as multicopper oxidases. This is the first report of an efficient BA reduction in wine by LAB. Furthermore, the identity of the enzymes involved has been revealed. PMID:23515835

  2. Marek's disease virus (MDV) ubiquitin-specific protease (USP) performs critical functions beyond its enzymatic activity during virus replication.

    Science.gov (United States)

    Veiga, Inês B; Jarosinski, Keith W; Kaufer, Benedikt B; Osterrieder, Nikolaus

    2013-03-15

    Marek's disease virus (MDV) encodes an ubiquitin-specific protease (USP) within its UL36 gene. USP is highly conserved among herpesviruses and was shown to be important for MDV replication and pathogenesis in MDV's natural host, the chicken. To further investigate the role of MDV USP, several recombinant (r) MDVs were generated and their in vitro phenotypes were evaluated using plaque size and growth kinetics assays. We discovered that the N-terminus of pUL36 is essential for MDV replication and could not be complemented by ectopic expression of MDV USP. In addition, we demonstrated that the region located between the conserved glutamine (Q85) and leucine (L106) residues comprising the active site cysteine (C98) is also essential for MDV replication. Based on the analyses of the rMDVs generated here, we concluded that MDV USP likely contributes to the structure and/or stability of pUL36 and affects replication and oncogenesis of MDV beyond its enzymatic activity. PMID:23399034

  3. Cell-free extracellular enzymatic activity is linked to seasonal temperature changes: a case study in the Baltic Sea

    Science.gov (United States)

    Baltar, Federico; Legrand, Catherine; Pinhassi, Jarone

    2016-05-01

    Extracellular enzymatic activities (EEAs) are a crucial step in the degradation of organic matter. Dissolved (cell-free) extracellular enzymes in seawater can make up a significant contribution of the bulk EEA. However, the factors controlling the proportion of dissolved EEA in the marine environment remain unknown. Here we studied the seasonal changes in the proportion of dissolved relative to total EEA (of alkaline phosphatase (APase), β-glucosidase (BGase), and leucine aminopeptidase (LAPase)), in the Baltic Sea for 18 months. The proportion of dissolved EEA ranged between 37 and 100, 0 and 100, and 34 and 100 % for APase, BGase, and LAPase, respectively. A consistent seasonal pattern in the proportion of dissolved EEA was found among all the studied enzymes, with values up to 100 % during winter and hydrolysis rates from microbial dynamics in cold waters. This implies that under cold conditions, cell-free enzymes can contribute to substrate availability at large distances from the producing cell, increasing the dissociation between the hydrolysis of organic compounds and the actual microbes producing the enzymes. This might also suggest a potential effect of global warming on the hydrolysis of organic matter via a reduction of the contribution of cell-free enzymes to the bulk hydrolytic activity.

  4. Pyridoxine Supplementation Improves the Activity of Recombinant Glutamate Decarboxylase and the Enzymatic Production of Gama-Aminobutyric Acid

    Science.gov (United States)

    Huang, Yan; Su, Lingqia; Wu, Jing

    2016-01-01

    Glutamate decarboxylase (GAD) catalyzes the irreversible decarboxylation of L-glutamate to the valuable food supplement γ-aminobutyric acid (GABA). In this study, GAD from Escherichia coli K12, a pyridoxal phosphate (PLP)-dependent enzyme, was overexpressed in E. coli. The GAD produced in media supplemented with 0.05 mM soluble vitamin B6 analog pyridoxine hydrochloride (GAD-V) activity was 154.8 U mL-1, 1.8-fold higher than that of GAD obtained without supplementation (GAD-C). Purified GAD-V exhibited increased activity (193.4 U mg-1, 1.5-fold higher than that of GAD-C), superior thermostability (2.8-fold greater than that of GAD-C), and higher kcat/Km (1.6-fold higher than that of GAD-C). Under optimal conditions in reactions mixtures lacking added PLP, crude GAD-V converted 500 g L-1 monosodium glutamate (MSG) to GABA with a yield of 100%, and 750 g L-1 MSG with a yield of 88.7%. These results establish the utility of pyridoxine supplementation and lay the foundation for large-scale enzymatic production of GABA. PMID:27438707

  5. Radiation modulation of the activity of some enzymatic systems of isolated plasma membranes in early ontogenesis

    International Nuclear Information System (INIS)

    Studied is the activity of adenilate cyclase (ADC-ase), phosphodiesterase tsAMF(FDEh) and 5'-nucleotidase of plasma membranes of liver extracted from rat embryo of the 20th day of development in norm and after the action of ionizing radiation. It is shown that γ-radiation of plasma membranes in doses from 0.1 to 100 kR decreases the ADCase activity with the more pronounced effect in the case of stimulation of isoproterenole under the conditions of large doses. The 5' nucleotidase and FDEh activity has not been changed up to 100 kR

  6. Integrin-mediated adhesion as self-sustained waves of enzymatic activation.

    Science.gov (United States)

    Block, M R; Destaing, O; Petropoulos, C; Planus, E; Albigès-Rizo, C; Fourcade, B

    2015-10-01

    Integrin receptors mediate interaction between the cellular actin-cytoskeleton and extracellular matrix. Based on their activation properties, we propose a reaction-diffusion model where the kinetics of the two-state receptors is modulated by their lipidic environment. This environment serves as an activator variable, while a second variable plays the role of a scaffold protein and controls the self-sustained activation of the receptors. Due to receptor diffusion which couples dynamically the activator and the inhibitor, our model connects major classes of reaction diffusion systems for excitable media. Spot and rosette solutions, characterized by receptor clustering into localized static or dynamic structures, are organized into a phase diagram. It is shown that diffusion and kinetics of receptors determines the dynamics and the stability of these structures. We discuss this model as a precursor model for cell signaling in the context of podosomes forming actoadhesive metastructures, and we study how generic signaling defects influence their organization. PMID:26565269

  7. beta-carotene does not change markers of enzymatic and nonenzymatic antioxidant activity in human blood

    DEFF Research Database (Denmark)

    Castenmiller, J.J.M.; Lauridsen, Søren T.; Dragsted, Lars O.; Hof, K.H.V.; Linssen, J.P.H.; West, C.E.

    1999-01-01

    = 12) or with a spinach product (n = 12 per group), i.e., whole-leaf, minced, liquefied or liquefied spinach plus added dietary fiber. After 3 wk of dietary intervention, changes in serum or plasma concentrations of ascorbic acid, alpha-tocopherol, FRAP (ferric reducing ability of plasma) and uric acid...... and erythrocyte enzyme activities were assessed, and differences among experimental groups were tested. Consumption of spinach resulted in greater (P <0.01) erythrocyte glutathione reductase activity and lower (P <0.05) erythrocyte catalase activity and serum alpha-tocopherol concentration compared...... with the control group. Consumption of the carotenoid supplement led to lower alpha-tocopherol responses (P = 0.02) compared with the basic diet only. Our data suggest that the short-term changes in erythrocyte glutathione reductase activity and serum alpha-tocopherol concentration can be attributed to...

  8. Antioxidant activity of camel milk casein before and after in vitro simulated enzymatic digestion

    OpenAIRE

    Zeineb Jrad; Jean-Michel Girardet; Isabelle Adt; Nadia Oulahal; Pascal Degraeve; Touhami Khorchani

    2014-01-01

    The effect of a successive in vitro hydrolysis by pepsin and pancreatin on the free radical scavenging activity of camel milk casein was investigated in order to assess the effect of gastro-intestinal digestion. Hydrolysis of camel casein was controlled by reversed-phase high performance liquid chromatography. Anti-oxidant activity was measured by the 2,2’-azino-bis-(3-ethylbensothiazoline-6- sulfonic acid) (ABTS) method. The Trolox equivalent antioxidant capacity (TEAC) values of camel casei...

  9. Spatial distribution of total phenolic content, enzymatic activities and browning in white yam (Dioscorea rotundata) tubers

    OpenAIRE

    Graham-Acquaah, Seth; Ayernor, George Sodah; Bediako-Amoa, Betty; Saalia, Firibu Kwesi; Afoakwa, Emmanuel Ohene

    2012-01-01

    Browning in raw and processed yams resulting from enzymes, polyphenol oxidase (PPO) and peroxidase (POD), activities is a major limitation to the industrial utilization of Dioscorea varieties of yams. Two elite cultivars of D. rotundata species were selected to study the spatial distribution of total phenols and enzymes (PPO and POD) activities. The intensities of tissue darkening in fresh yam chips prepared from the tuber sections of cultivars during frozen storage were also studied. Total p...

  10. Irradiation effect on enzymatic activity of papain with {sup 60}Co-{gamma} rays

    Energy Technology Data Exchange (ETDEWEB)

    Furuta, Masakazu; Ohashi, Isao; Oka, Masahito; Hayashi, Toshio [Osaka Prefecture Univ., Sakai (Japan). Research Inst. for Advanced Science and Technology

    1998-12-31

    An investigation was made on the durability of enzyme activity against {sup 60}Co-{gamma} irradiation at a dose up to 55 kGy/h using dry powder and aqueous solution of papain preparations on the market. Hybrid materials including bioactive molecules combined with biocompatible synthetic polymers are expected to have biocompatible properties and also biomimetic functions as a component of artificial organs for human body. The activity of papain in an aqueous solution was rapidly decreased at the early stage of irradiation through oxidation of SH group at its active site with active oxygen produced by the irradiation and then, partially recovered since SH group was reproduced in an anoxic state after O{sub 2} consumption in the solution irradiated at a high dose. A usual radiation method for sterilization was found applicable to decontamination of dry and frozen preparations of papain. When suitable conditions for radiation were chosen and N{sub 2} gas was purged to suppress the formation of free radicals, it was possible to keep the enzyme activity at more than 50% of the initial activity after radiation at 30 kGy. (M.N.)

  11. Irradiation effect on enzymatic activity of papain with 60Co-γ rays

    International Nuclear Information System (INIS)

    An investigation was made on the durability of enzyme activity against 60Co-γ irradiation at a dose up to 55 kGy/h using dry powder and aqueous solution of papain preparations on the market. Hybrid materials including bioactive molecules combined with biocompatible synthetic polymers are expected to have biocompatible properties and also biomimetic functions as a component of artificial organs for human body. The activity of papain in an aqueous solution was rapidly decreased at the early stage of irradiation through oxidation of SH group at its active site with active oxygen produced by the irradiation and then, partially recovered since SH group was reproduced in an anoxic state after O2 consumption in the solution irradiated at a high dose. A usual radiation method for sterilization was found applicable to decontamination of dry and frozen preparations of papain. When suitable conditions for radiation were chosen and N2 gas was purged to suppress the formation of free radicals, it was possible to keep the enzyme activity at more than 50% of the initial activity after radiation at 30 kGy. (M.N.)

  12. Modulation of the captopril interference with the activity of some enzymatic biomolecules in monkey kidney vero cells by drug delivery mesoporous silica system

    Directory of Open Access Journals (Sweden)

    Roxana Popovici

    2011-12-01

    Full Text Available The in vitro effect of different formulations of captopril on some cellular enzymatic equipments activities of monkey kidney Vero cells was investigated in the present research. The preparation of the samples of the mesoporous silica nanocomposites, loaded or not with captopril, was described and their effect on membranary Na+-K+-ATP-ase, cell Mg2+-ATP-ase, LDH, Px, GSH-Px, SOD, CAT, ACP, ALP activities were studied. The Vero cells were incubated, for a period of 144 hours, with growing medium renewed twice. When the cells reached confluence in the monolayer stage, the cultures were divided into control cell cultures and other 4 treated groups. To the 12 hours treated cells were added: Cap H2, SBA–15, unfunctionalized SBA-15_CapH2_RT and functionalized SBA-15_APTES_CapH2_80°C nanocomposites, each of them in a dose of 0.4μg./flask. As compared with the control Vero cells, which are characterized by a specific level of the enzymatic activities, the cultures treated with SBA-15 have not presented significant alterations of them. The comparative study of captopril interactions with some membrane bound and intracellular enzymatic biomolecules of monkey kidney Vero cells has revealed either an enhancement of membranary Na+-K+-ATP-ase, intracell total ATP- ase , LDH, ACP , and GSH-Px activities or a repression of cellular CAT, Px and SOD activities. These variations of the enzymatic activities – which induce modifications of the membranary and metabolic processes – could be due to a direct or indirect interaction of captopril with cellular (plasmalemma or subcellular (organelles structures and with intracellular biomolecules (enzymes, DNA, RNA etc.. The association of captoptil with SBA – 15 or SBA – 15 _ APTES mesoporous silica matrices and treatment of Vero cells with these nanocomposites were correlated with modulation of the captopril interference with the activity of investigated enzymatic biomolecules, its sense (stimulation or

  13. Two-Photon Enzymatic Probes Visualizing Sub-cellular/Deep-brain Caspase Activities in Neurodegenerative Models.

    Science.gov (United States)

    Qian, Linghui; Zhang, Cheng-Wu; Mao, Yanli; Li, Lin; Gao, Nengyue; Lim, Kah-Leong; Xu, Qing-Hua; Yao, Shao Q

    2016-01-01

    Caspases work as a double-edged sword in maintaining cell homeostasis. Highly regulated caspase activities are essential during animal development, but dysregulation might lead to different diseases, e.g. extreme caspase activation is known to promote neurodegeneration. At present, visualization of caspase activation has mostly remained at the cellular level, in part due to a lack of cell-permeable imaging probes capable of direct, real-time investigations of endogenous caspase activities in deep tissues. Herein, we report a suite of two-photon, small molecule/peptide probes which enable sensitive and dynamic imaging of individual caspase activities in neurodegenerative models under physiological conditions. With no apparent toxicity and the ability of imaging endogenous caspases both in different subcellular organelles of mammalian cells and in brain tissues, these probes serve as complementary tools to conventional histological analysis. They should facilitate future explorations of caspases at molecular, cellular and organism levels and inspire development of novel two-photon probes against other enzymes. PMID:27210613

  14. ACE-inhibitory activity of enzymatic protein hydrolysates from lupin and other legumes.

    Science.gov (United States)

    Boschin, Giovanna; Scigliuolo, Graziana Maria; Resta, Donatella; Arnoldi, Anna

    2014-02-15

    The objective of this investigation was to compare the angiotensin converting enzyme (ACE)-inhibitory activity of the hydrolysates obtained by pepsin digestion of proteins of some legumes, such as chickpea, common bean, lentil, lupin, pea, and soybean, by using the same experimental procedure. The ACE-inhibitory activity was measured by using the tripeptide hippuryl-histidyl-leucine (HHL), as model peptide, and HPLC-DAD, as analytical method. The peptide mixtures of all legumes were active, with soybean and lupin the most efficient, with IC50 values of 224 and 226 μg/ml, respectively. Considering the promising results obtained with lupin, and aiming to identify the protein(s) that release(s) the peptides responsible for the activity, the peptides obtained from the pepsin digestion of some industrial lupin protein isolates and purified protein fractions were tested. The most active mixture, showing an IC50 value of 138 μg/ml, was obtained hydrolysing a mixture of lupin α+β conglutin. PMID:24128446

  15. Bacterial Enzymatic Activity and Bioavailability of Heavy Metals in Sediments from Boa Viagem Beach (Guanabara Bay

    Directory of Open Access Journals (Sweden)

    Mirian Crapez

    2003-01-01

    Full Text Available This study focuses on the quality of the organic matter that reaches the sediment from Boa Viagem Beach and through the evaluation of the total bacterial count, the electron transport system activity (ETSA, the esterase activity (EST, as well as the protein and the organic matter contents. Seasonal variations of organic matter, protein content and the number of bacteria were particularly notable in the summer. ETSA reached a maximum of 7.48 µl O 2 h-1 g-1 in the summer. EST activity presented a different pattern once it reached a maximum of 0.17 µg fluorescein h-1 g-1 in the winter. The temporal variation of ETSA and EST activity indicated that biopolymers predominated in the winter, and oligomers or monomers predominated in the summer. These results suggest that organic carbon turnover is more likely to be controlled by organic matter quality. The heavy metals concentrations, especially for Cu, Zn, Ni and Cr, indicated absence of the inhibition of dehydrogenase activity, and they are not bioavailable in the EC 50 values

  16. Determination of antioxidant enzymatic activity in several halophytes from Dobrogea area

    Directory of Open Access Journals (Sweden)

    Maria Aurelia Ivan

    2012-10-01

    Full Text Available Halophytes have evolved various mechanisms of adaptations to stress tolerance including anincrease of antioxidant enzymes activities. The present study was conducted in order to investigate the activity of someantioxidant enzymes (superoxide dismutase – SOD, catalase – CAT and peroxidase – POD in several halophytes. Forthis, four halophytic species were collected during summer of 2012 from two distinct saline areas, located in South-Eastof Romania (Dobrogea. Species collected from Histria are Plantago maritima and Bassia sedoides; the first mentionedspecies was collected in various stages of development (vegetative and flowering phases. Species Plantago coronopus, Spergularia media, Limonium gmelini, and Bassia sedoides from Sulina were collected from two different habitats: littoral area and an habitat located at 1000 m from littoral.The results show that halophytes collected from 1000 m to littoral area were characterized by higher levels ofSOD activity than those collected from littoral. The peroxidase activity in halophytes collected from Sulina show variousresponses according to species and collecting points. Some of halophytes collected from Histria and Sulina have anundetectable level of catalase activity at the moment of determination; perhaps the role of this enzyme for removing H2O2 has been taken by peroxidase.

  17. Role of trypsin-like cleavage at arginine 192 in the enzymatic and cytotonic activities of Escherichia coli heat-labile enterotoxin.

    Science.gov (United States)

    Grant, C C; Messer, R J; Cieplak, W

    1994-01-01

    Previous studies of cholera toxin and Escherichia coli heat-labile enterotoxin have suggested that proteolytic cleavage plays an important role in the expression of ADP-ribosyltransferase activity and toxicity. Specifically, several studies have implicated a trypsin-like cleavage at arginine 192, which lies within an exposed region subtended by a disulfide bond in the intact A subunit, in toxicity. To investigate the role of this modification in the enzymatic and cytotonic properties of heat-labile enterotoxin, the response of purified, recombinant A subunit to tryptic activation and the effect of substituting arginine 192 with glycine on the activities of the holotoxin were examined. The recombinant A subunit of heat-labile enterotoxin exhibited significant levels of ADP-ribosyltransferase activity that were only nominally increased (approximately twofold) by prior limited trypsinolysis. The enzymatic activity also did not appear to be affected by auto-ADP-ribosylation that occurs during the high-level synthesis of the recombinant A subunit in E. coli. A mutant form of the holotoxin containing the arginine 192-to-glycine substitution exhibited levels of cytotonic activity for CHO cells that were similar to that of the untreated, wild-type holotoxin but exhibited a marked delay in the ability to increase intracellular levels of cyclic AMP in Caco-2 cells. The results indicate that trypsin-like cleavage of the A subunit of E. coli heat-labile enterotoxin at arginine 192 is not requisite to the expression of enzymatic activity by the A subunit and further reveal that this modification, although it enhances the biological and enzymatic activities of the toxin, is not absolutely required for the enterotoxin to elicit cytotonic effects. Images PMID:7927684

  18. Antioxidant activity of camel milk casein before and after in vitro simulated enzymatic digestion

    Directory of Open Access Journals (Sweden)

    Zeineb Jrad

    2014-11-01

    Full Text Available The effect of a successive in vitro hydrolysis by pepsin and pancreatin on the free radical scavenging activity of camel milk casein was investigated in order to assess the effect of gastro-intestinal digestion. Hydrolysis of camel casein was controlled by reversed-phase high performance liquid chromatography. Anti-oxidant activity was measured by the 2,2’-azino-bis-(3-ethylbensothiazoline-6- sulfonic acid (ABTS method. The Trolox equivalent antioxidant capacity (TEAC values of camel casein and its hydrolysate were 1.6±0.12 μmol TE/mg protein and 0.25 μmol TE/μmol eq. NH2, respectively. After digestion, the scavenging activity of the casein peptides was more efficient than those reported in the literature regarding digestive hydrolysates of camel milk, colostrum and whey proteins.

  19. Effect of Static Magnetic Field on α-Amylase Activity and Enzymatic Reaction

    Institute of Scientific and Technical Information of China (English)

    JIA Shaoyi; LIU Yong; WU Songhai; WANG Zhibin

    2009-01-01

    The effect of magnetic field on α-amylase was studied. Under the experimental conditions, α-amylase solution was treated by 0.15 T, 0.30 T and 0.45 T static magnetic fields for a known period of time, then the activ-ity, kinetic parameters, and the secondary conformation were investigated. The results showed that there was a con-siderable effect of the magnetic exposure on the α-amylase. The activity was increased by 27%, 34.1%, 37.8% compared with the control. It was also found that both kinetic parameters Km and Vm could be decreased due to the increasing magnetic field, Km decreased from 2.20×102 to 0.87×102, whereas Vm decreased from 2.0×103 g/min to 1.1×103g/min. At the same time, there were some irregular changes in α-amylase secondary conformation.

  20. Enzymatic activities of the GB virus-B RNA-dependent RNA polymerase

    International Nuclear Information System (INIS)

    The GB virus-B (GBV-B) nonstructural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp) with greater than 50% sequence similarity to the hepatitis C virus (HCV) NS5B. Recombinant GBV-B NS5B was reported to possess RdRp activity (W. Zhong et al., 2000, J. Viral Hepat. 7, 335-342). In this study, the GBV-B RdRp was examined more thoroughly for different RNA synthesis activities, including primer-extension, de novo initiation, template switch, terminal nucleotide addition, and template specificity. The results can be compared with previous characterizations of the HCV RdRp. The two RdRps share similarities in terms of metal ion and template preference, the abilities to add nontemplated nucleotides, perform both de novo initiation and extension from a primer, and switch templates. However, several differences in RNA synthesis between the GBV-B and HCV RdRps were observed, including (i) optimal temperatures for activity, (ii) ranges of Mn2+ concentration tolerated for activity, and (iii) cation requirements for de novo RNA synthesis and terminal transferase activity. To assess whether the recombinant GBV-B RdRp may represent a relevant surrogate system for testing HCV antiviral agents, two compounds demonstrated to be active at nanomolar concentrations against HCV NS5B were tested on the GBV RdRp. A chain terminating nucleotide analog could prevent RNA synthesis, while a nonnucleoside HCV inhibitor was unable to affect RNA synthesis by the GBV RdRp

  1. Certain enzymatic activities in brain and liver mitochondria of rats treated with pantothenic acid after irradiation

    International Nuclear Information System (INIS)

    Whole body caesium-137 gamma irradiation of rats with single dose of 5 Gy induced significant decrease in the activities of glutamate dehydrogenase, isocitrate dehydrogenase and succunate dehydrogenase in mitochondria of brain and liver. Intraperitoneal administration of pantothenic acid (20 mg/Kg body weight/day) for 5 consecutive days after irradiation resulted of detectable improvement in the radiation-induced decrease inactivities of mitochondrial enzymes. It is postulated that pantothenic acid administered to rats after irradiation might play a role in the regulation of certain mitochondrial enzymes activities

  2. Secretory Phospholipases A2 in Durum Wheat (Triticum durum Desf.: Gene Expression, Enzymatic Activity, and Relation to Drought Stress Adaptation

    Directory of Open Access Journals (Sweden)

    Daniela Trono

    2013-03-01

    Full Text Available Phospholipases A2 (PLA2s are known to mediate signaling cascades during plant growth and development, as well as biotic and abiotic stress responses. In this context, the present study provides extensive characterization of specific PLA2s in durum wheat, and assesses their involvement in durum wheat response to drought stress. In durum wheat leaves, four full-length expressed sequences encoding putative PLA2s were isolated and characterized as belonging to the class of secretory PLA2s (sPLA2s: TdsPLA2I, TdsPLA2II, TdsPLA2III and TdsPLA2IV. PLA2 activity was also detected, the characteristics of which resemble those of previously characterized plant sPLA2s: strong preference for phospholipids; requirement for millimolar Ca2+ concentrations; optimal activity at basic pH; heat stability; and inhibition by the reducing agent dithiothreitol. With drought stress imposed at both the vegetative and reproductive stages, accumulation of TdsPLA2I and TdsPLA2III transcripts, and to a lesser extent of TdsPLA2IV transcript, paralleled increased PLA2 activity; both transcript levels and enzymatic activity decreased as a consequence of stress recovery. Consistently, free fatty acid analysis of drought-stressed leaves revealed increased linoleate, linolenate and palmitate contents, which were reversed by plant re-watering. Overall, these findings strongly suggest that there are inducible sPLA2 isoforms in durum wheat that have roles in orchestrating the plant response to drought stress.

  3. Iron(II) Supramolecular Helicates Condense Plasmid DNA and Inhibit Vital DNA-Related Enzymatic Activities

    Czech Academy of Sciences Publication Activity Database

    Malina, Jaroslav; Hannon, M.J.; Brabec, Viktor

    2015-01-01

    Roč. 21, č. 31 (2015), s. 11189-11195. ISSN 0947-6539 R&D Projects: GA ČR(CZ) GA13-08273S Institutional support: RVO:68081707 Keywords : TOPOISOMERASE-I * METALLOSUPRAMOLECULAR CYLINDERS * TRANSCRIPTIONAL ACTIVITY Subject RIV: BO - Biophysics Impact factor: 5.731, year: 2014

  4. Insecticidal effects of Moroccan plant extracts on development, energy reserves and enzymatic activities of Plodia interpunctella

    Energy Technology Data Exchange (ETDEWEB)

    Bouayard, N.; Rharrabe, K.; Ghailani, N. N.; Jbilou, R.; Castanera, P.; Ortego, F.

    2013-05-01

    The aim of this work was to study the effects of methanol extracts of ten plant species used in traditional medicine in Morocco (Peganum harmala, Ajuga iva, Rosmarinus officinalis, Lavandula stoechas, Lavandula dentata, Cistus ladanifer, Cistus salviaefolius, Cistus monspeliensis, Centaurium erythraea and Launaea arborescens) on Plodia interpunctella Hubner (Lepidoptera: Pyralidae) larvae. Firstly, we studied the effects of the ingestion of these extracts at 500 ppm on post-embryonic development parameters. Most plant extracts provoked a notable decrease of larval weight 8 days after treatment (up to 33% weight loss with C. erythraea) and caused significant alterations on pupation (ranging from 5% to 85%) and adult emergence (below 2.5% with R. officinalis, C. erythraea and A. iva). The plant extracts that showed strongest effects on post-embryonic development were selected to test their effects on the following physiological parameters: larval reserve substances (at 500 ppm); and midgut activities of hydrolytic and detoxification enzymes (at 500, 750 and 1000 ppm). All treatments provoked a significant reduction of protein and carbon hydrate larval contents, the inhibition of proteases and {alpha}-amylase activities in a dose depended manner, and the induction of glutathione S-transferase and esterase (using MtB as substrate) activities, whereas the activity of cytochrome P450 monooxygenases and esterases (using 1-NA as substrate) increase or decrease depending on the extract concentration and the plant analyzed. (Author) 65 refs.

  5. Chemical Characterization, Antioxidant and Enzymatic Activity of Brines from Scandinavian Marinated Herring Products

    DEFF Research Database (Denmark)

    Gringer, Nina; Osman, Ali; Nielsen, Henrik Hauch; Undeland, Ingrid; Baron, Caroline P.

    2014-01-01

    Brines generated during the last marination step in the production of marinated herring (Clupea harengus) were chemically characterized and analyzed for antioxidant and enzyme activities. The end-products were vinegar cured, spice cured and traditional barrel-salted herring with either salt or...

  6. Periplasmic nitrate reductase and formate dehydrogenase: similar molecular architectures with very different enzymatic activities.

    Science.gov (United States)

    Cerqueira, Nuno M F S A; Gonzalez, Pablo J; Fernandes, Pedro A; Moura, José J G; Ramos, Maria João

    2015-11-17

    It is remarkable how nature has been able to construct enzymes that, despite sharing many similarities, have simple but key differences that tune them for completely different functions in living cells. Periplasmic nitrate reductase (Nap) and formate dehydrogenase (Fdh) from the DMSOr family are representative examples of this. Both enzymes share almost identical three-dimensional protein foldings and active sites, in terms of coordination number, geometry and nature of the ligands. The substrates of both enzymes (nitrate and formate) are polyatomic anions that also share similar charge and stereochemistry. In terms of the catalytic mechanism, both enzymes have a common activation mechanism (the sulfur-shift mechanism) that ensures a constant coordination number around the metal ion during the catalytic cycle. In spite of these similarities, they catalyze very different reactions: Nap abstracts an oxygen atom from nitrate releasing nitrite, whereas FdH catalyzes a hydrogen atom transfer from formate and releases carbon dioxide. In this Account, a critical analysis of structure, function, and catalytic mechanism of the molybdenum enzymes periplasmic nitrate reductase (Nap) and formate dehydrogenase (Fdh) is presented. We conclude that the main structural driving force that dictates the type of reaction, catalyzed by each enzyme, is a key difference on one active site residue that is located in the top region of the active sites of both enzymes. In both enzymes, the active site is centered on the metal ion of the cofactor (Mo in Nap and Mo or W in Fdh) that is coordinated by four sulfur atoms from two pyranopterin guanosine dinucleotide (PGD) molecules and by a sulfido. However, while in Nap there is a Cys directly coordinated to the Mo ion, in FdH there is a SeCys instead. In Fdh there is also an important His that interacts very closely with the SeCys, whereas in Nap the same position is occupied by a Met. The role of Cys in Nap and SeCys in FdH is similar in both

  7. The effects of mediator and granular activated carbon addition on degradation of trace organic contaminants by an enzymatic membrane reactor.

    Science.gov (United States)

    Nguyen, Luong N; Hai, Faisal I; Price, William E; Leusch, Frederic D L; Roddick, Felicity; Ngo, Hao H; Guo, Wenshan; Magram, Saleh F; Nghiem, Long D

    2014-09-01

    The removal of four recalcitrant trace organic contaminants (TrOCs), namely carbamazepine, diclofenac, sulfamethoxazole and atrazine by laccase in an enzymatic membrane reactor (EMR) was studied. Laccases are not effective for degrading non-phenolic compounds; nevertheless, 22-55% removal of these four TrOCs was achieved by the laccase EMR. Addition of the redox-mediator syringaldehyde (SA) to the EMR resulted in a notable dose-dependent improvement (15-45%) of TrOC removal affected by inherent TrOC properties and loading rates. However, SA addition resulted in a concomitant increase in the toxicity of the treated effluent. A further 14-25% improvement in aqueous phase removal of the TrOCs was consistently observed following a one-off dosing of 3g/L granular activated carbon (GAC). Mass balance analysis reveals that this improvement was not due solely to adsorption but also enhanced biodegradation. GAC addition also reduced membrane fouling and the SA-induced toxicity of the effluent. PMID:24980029

  8. Investigation of trypsin-CdSe quantum dot interactions via spectroscopic methods and effects on enzymatic activity

    Science.gov (United States)

    Kaur, Gurvir; Tripathi, S. K.

    2015-01-01

    The paper presents the interactions between trypsin and water soluble cadmium selenide (CdSe) quantum dots investigated by spectrophotometric methods. CdSe quantum dots have strong ability to quench the intrinsic fluorescence of trypsin by a static quenching mechanism. The quenching has been studied at three different temperatures where the results revealed that electrostatic interactions exist between CdSe quantum dots and trypsin and are responsible to stabilize the complex. The Scatchard plot from quenching revealed 1 binding site for quantum dots by trypsin, the same has been confirmed by making isothermal titrations of quantum dots against trypsin. The distance between donor and acceptor for trypsin-CdSe quantum dot complexes is calculated to be 2.8 nm by energy transfer mechanisms. The intrinsic fluorescence of CdSe quantum dots has also been enhanced by the trypsin, and is linear for concentration of trypsin ranging 1-80 μl. All the observations evidence the formation of trypsin-CdSe quantum dot conjugates, where trypsin retains the enzymatic activity which in turn is temperature and pH dependent.

  9. Differential coral bleaching-Contrasting the activity and response of enzymatic antioxidants in symbiotic partners under thermal stress.

    Science.gov (United States)

    Krueger, Thomas; Hawkins, Thomas D; Becker, Susanne; Pontasch, Stefanie; Dove, Sophie; Hoegh-Guldberg, Ove; Leggat, William; Fisher, Paul L; Davy, Simon K

    2015-12-01

    Mass coral bleaching due to thermal stress represents a major threat to the integrity and functioning of coral reefs. Thermal thresholds vary, however, between corals, partly as a result of the specific type of endosymbiotic dinoflagellate (Symbiodinium sp.) they harbour. The production of reactive oxygen species (ROS) in corals under thermal and light stress has been recognised as one mechanism that can lead to cellular damage and the loss of their symbiont population (Oxidative Theory of Coral Bleaching). Here, we compared the response of symbiont and host enzymatic antioxidants in the coral species Acropora millepora and Montipora digitata at 28°C and 33°C. A. millepora at 33°C showed a decrease in photochemical efficiency of photosystem II (PSII) and increase in maximum midday excitation pressure on PSII, with subsequent bleaching (declining photosynthetic pigment and symbiont density). M. digitata exhibited no bleaching response and photochemical changes in its symbionts were minor. The symbiont antioxidant enzymes superoxide dismutase, ascorbate peroxidase, and catalase peroxidase showed no significant upregulation to elevated temperatures in either coral, while only catalase was significantly elevated in both coral hosts at 33°C. Increased host catalase activity in the susceptible coral after 5days at 33°C was independent of antioxidant responses in the symbiont and preceded significant declines in PSII photochemical efficiencies. This finding suggests a potential decoupling of host redox mechanisms from symbiont photophysiology and raises questions about the importance of symbiont-derived ROS in initiating coral bleaching. PMID:26310104

  10. Enzymatic activity of soluble and membrane tethered peptide pro-hormone convertase 1.

    Science.gov (United States)

    Bruzzaniti, Angela; Mains, Richard E

    2002-05-01

    Pro-hormone convertases PC1 and PC2 perform endoproteolytic cleavages of precursors in peptide-containing secretory granules. PC1 and PC2 are soluble, secreted with bioactive peptides. Evolutionarily related PCs have membrane tethers, not secreted. We tethered PC1 to the transmembrane-cytoplasmic domains (CD) of a granule enzyme (peptidylglycine-alpha-amidating monooxygenase; PAM) and Golgi-localized PC8. The tethered PC1 is far more stable to elevated temperature and denaturants than soluble PC1, and more active. Both tethers allow PC1 to visit the cell surface transiently, cleaving soluble molecules outside the cell. Both membrane-bound PC1 chimeras cleave membrane PAM into soluble active fragments when PAM is expressed on adjacent cells. PMID:12084516

  11. Gamma irradiation effect on the enzymatic activities of horseradish and apple peroxidases

    International Nuclear Information System (INIS)

    The behavior at low-dose exposure (0.033-0.4 kGy) of horseradish peroxidase (HRP) and of two different purified fractions of apple (Jonathan cultivar) peroxidases (named APR1S and APR2S) was studied. The HRP solutions were added with either 0.32 M fructose or glucose in order to study their effect on enzymes activity response under γ (137Cs, dose rate 0.4 kGy/h) irradiation. The obtained results showed similar behavior between HRP-sugar-added solution and apple fraction with higher oligosaccharides content (APR2S) undergoing low-dose treatment. The same pattern was observed between unglycosylated HRP and APR1S with lower oligosaccharides content. These similarities gave us the possibility to conclude that the presence of oligosaccharides, in more or less quantities, influences in the same way the peroxidases activity, from different plant species, exposed to γ irradiation

  12. [Study of the Sporothrix schenkii (yeast forms) extract. Electrophoretic and immunoelectrophoretic analyses: characterization of enzymatic activities].

    Science.gov (United States)

    Walbaum, S; Duriez, T; Dujardin, L; Biguet, J

    1978-07-28

    An extract from living yeast forms of S. schenckii was prepared. The yeasts originated from a shake culture in B.H.I. broth (Difco) incubated for 3 days at 35 degrees C in darkness; they were harvested, washed and disrupted with glass beads in a model MSK Braun mechanical cell homogenizer; a freezing-thawing was added to improve the extract. After electrophoretic separation in agarose gel, the extract's components were characterized by their enzymic activity; with this technique, 30 bands were revealed. These enzymic activities were also investigated on the antigenic fractions of the extract revealed by a rabbit hyperimmunserum: 16 among 22 immunoprecipitates are identified by their catalytic properties. Study of the earliest precipitating antibodies (appearing-order and enzymic caracterization) in rabbits just immunized completes this work. How to ameliorate the quality of the extract by culture and extraction conditions is also specified. PMID:692628

  13. Characterisation of enzymatic activities of H5N1 influenza virus

    Directory of Open Access Journals (Sweden)

    Simson Tarigan

    2008-06-01

    Full Text Available One of the two glycoproteins projected from the surface of the influenza virus is identified as neuraminidase. This enzyme enables the virus to spread in the host, and therefore it plays vital roles in the viral pathogenicity. From the viewpoint of disease control, neuraminidase is used as the target for the development of anti-flu drugs, and for the development of diagnostic test to differentiate infected from vaccinated animals (DIVA. Since the roles of the enzyme are very important, information regarding the characteristics and the procedure to measure its activity, which is the purpose of this study, is essential. The optimum incubation time of the neuraminidase-substrate (fetuin reaction and the optimum pH of the buffer were determined. The stability of the enzyme against heating, supplementation or chelating of calcium ion, and b-propiolactone treatment were analysed. This study showed that neuraminidase from H5N1-influenza virus was, in regards to the characteristics investigated in this study, was comparable to that from Clostridium perfringens. The optimum incubation time for the viral and Clostridial neuraminidases were 60 and 30 minutes, respectively; whereas, the optimum pH for both neuraminidase was 6-7. At pH 8, both neuraminidase were inactive. Supplementation of calcium ion tended to increase activity but chelating of the cation did not have any observable effects. Treatment with 0.2% b-propiolactone for 6 hours reduced the activity, whereas heating at 60°C for 60 minutes abolished all activity. Since inactivation by b-propiolactone is partially only, neuraminidase assay could be performed safely in ordinary laboratories using b-propiolactone-treated-influenza virus, rather than the life virus. The thermolabile nature of the enzyme will complicate any attempt to purify the enzyme.

  14. Clustered mutations in hominid genome evolution are consistent with APOBEC3G enzymatic activity

    OpenAIRE

    Pinto, Yishay; Gabay, Orshay; Arbiza, Leonardo; Sams, Aaron J.; Keinan, Alon; Levanon, Erez Y.

    2016-01-01

    The gradual accumulation of mutations by any of a number of mutational processes is a major driving force of divergence and evolution. Here, we investigate a potentially novel mutational process that is based on the activity of members of the AID/APOBEC family of deaminases. This gene family has been recently shown to introduce—in multiple types of cancer—enzyme-induced clusters of co-occurring somatic mutations caused by cytosine deamination. Going beyond somatic mutations, we hypothesized t...

  15. Enzymatic activation of Fujinami sarcoma virus gag-fps transforming proteins by autophosphorylation at tyrosine.

    OpenAIRE

    Meckling-Hansen, K; Nelson, R; Branton, P; Pawson, T

    1987-01-01

    Site-directed mutagenesis of the Fujinami sarcoma virus (FSV) genome has suggested that Tyr 1073 of the P130gag--fps protein-tyrosine kinase is a regulatory site. To investigate directly the ability of tyrosine phosphorylation to affect P130gag--fps kinase activity, the phosphotyrosyl phosphatase inhibitor orthovanadate and partially purified phosphotyrosyl phosphatases were used to manipulate the stoichiometry of P130gag--fps phosphorylation. Phosphorylation of P130gag--fps at Tyr 1073 corre...

  16. Integrated catalysis opens new arylation pathways via regiodivergent enzymatic C–H activation

    OpenAIRE

    Latham, Jonathan; Henry, Jean-Marc; Sharif, Humera H.; Menon, Binuraj R. K.; Shepherd, Sarah A; Greaney, Michael F; Micklefield, Jason

    2016-01-01

    Despite major recent advances in C–H activation, discrimination between two similar, unactivated C–H positions is beyond the scope of current chemocatalytic methods. Here we demonstrate that integration of regioselective halogenase enzymes with Pd-catalysed cross-coupling chemistry, in one-pot reactions, successfully addresses this problem for the indole heterocycle. The resultant ‘chemobio-transformation' delivers a range of functionally diverse arylated products that are impossible to acces...

  17. Some Enzymatic Activities Of Fungi Isolated From Cotton Seeds And Cotton Seed Products

    OpenAIRE

    El Kady, I. A. [اسماعيل عبد الرزاق القاضي; Mazen, Mohamed B.; Saber, Sabeh M.

    1984-01-01

    Five hundred different cultures which belong to fifteen genera and forty-one species, isolated from cotton seeds and cotton seed products were studied for proteolytic, amylolytic and lipolytic activities. About 80% of the cultures tested proved to be protease-producers. High proportions of protease producing cultures were found in the genera Aspergillus (97.73%), Fusarium (95.83%) and Penicillium (46.34%). Most of the aspergilli which belong to the species A. flavus, A. fumigatus, A. sydowit ...

  18. A Redundant Role of Human Thyroid Peroxidase Propeptide for Cellular, Enzymatic, and Immunological Activity

    OpenAIRE

    Godlewska, Marlena; Góra, Monika; Buckle, Ashley M.; Porebski, Benjamin T.; Kemp, E. Helen; Sutton, Brian J.; Czarnocka, Barbara; Banga, J. Paul

    2014-01-01

    Background: Thyroid peroxidase (TPO) is a dimeric membrane-bound enzyme of thyroid follicular cells, responsible for thyroid hormone biosynthesis. TPO is also a common target antigen in autoimmune thyroid disease (AITD). With two active sites, TPO is an unusual enzyme, and thus there is much interest in understanding its structure and role in AITD. Homology modeling has shown TPO to be composed of different structural modules, as well as a propeptide sequence. During the course of studies to ...

  19. Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities

    International Nuclear Information System (INIS)

    The potential impact of six antibiotics (chlortetracycline, tetracycline and tylosin; sulfamethoxazole, sulfamethazine and trimethoprim) on plant growth and soil quality was studied by using seed germination test on filter paper and plant growth test in soil, soil respiration and phosphatase activity tests. The phytotoxic effects varied between the antibiotics and between plant species (sweet oat, rice and cucumber). Rice was most sensitive to sulfamethoxazole with the EC10 value of 0.1 mg/L. The antibiotics tested inhibited soil phosphatase activity during the 22 days' incubation. Significant effects on soil respiration were found for the two sulfonamides (sulfamethoxazole and sulfamethazine) and trimethoprim, whereas little effects were observed for the two tetracyclines and tylosin. The effective concentrations (EC10 values) for soil respiration in the first 2 days were 7 mg/kg for sulfamethoxazole, 13 mg/kg for sulfamethazine and 20 mg/kg for trimethoprim. Antibiotic residues in manure and soils may affect soil microbial and enzyme activities. - Terrestrial ecotoxicological effects of antibiotics are related to their sorption and degradation behavior in soil.

  20. Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities

    Energy Technology Data Exchange (ETDEWEB)

    Liu Feng [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Ying Guangguo, E-mail: guangguo.ying@gmail.co [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Tao Ran; Zhao Jianliang; Yang Jifeng [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Zhao Lanfeng [College of Resource and Environmental Science, South China Agricultural University, Guangzhou 510642 (China)

    2009-05-15

    The potential impact of six antibiotics (chlortetracycline, tetracycline and tylosin; sulfamethoxazole, sulfamethazine and trimethoprim) on plant growth and soil quality was studied by using seed germination test on filter paper and plant growth test in soil, soil respiration and phosphatase activity tests. The phytotoxic effects varied between the antibiotics and between plant species (sweet oat, rice and cucumber). Rice was most sensitive to sulfamethoxazole with the EC10 value of 0.1 mg/L. The antibiotics tested inhibited soil phosphatase activity during the 22 days' incubation. Significant effects on soil respiration were found for the two sulfonamides (sulfamethoxazole and sulfamethazine) and trimethoprim, whereas little effects were observed for the two tetracyclines and tylosin. The effective concentrations (EC10 values) for soil respiration in the first 2 days were 7 mg/kg for sulfamethoxazole, 13 mg/kg for sulfamethazine and 20 mg/kg for trimethoprim. Antibiotic residues in manure and soils may affect soil microbial and enzyme activities. - Terrestrial ecotoxicological effects of antibiotics are related to their sorption and degradation behavior in soil.

  1. Antimicrobial and antioxidant activities of clove essential oil and eugenyl acetate produced by enzymatic esterification.

    Science.gov (United States)

    Vanin, Adriana B; Orlando, Tainara; Piazza, Suelen P; Puton, Bruna M S; Cansian, Rogério L; Oliveira, Debora; Paroul, Natalia

    2014-10-01

    This work reports the maximization of eugenyl acetate production by esterification of essential oil of clove in a solvent-free system using Novozym 435 as catalyst. The antimicrobial and antioxidant activities of clove essential oil and eugenyl acetate produced were determined. The conditions that maximized eugenyl acetate production were 60 °C, essential oil of clove to acetic anhydride ratio of 1:5, 150 rpm, and 10 wt% of enzyme, with a conversion of 99.87 %. A kinetic study was performed to assess the influence of substrates' molar ratio, enzyme concentration, and temperature on product yield. Results show that an excess of anhydride, enzyme concentration of 5.5 wt%, 50 °C, and essential oil of clove to acetic anhydride ratio of 1:5 afforded nearly a complete conversion after 2 h of reaction. Comparing the antibacterial activity of the essential oil of clove before and after esterification, we observed a decrease in the antimicrobial activity of eugenyl acetate, particularly with regard to minimum inhibitory concentration (MIC). Both eugenyl acetate and clove essential oil were most effective to the gram-negative than gram-positive bacteria group. The results showed a high antioxidant potential for essential oil before and particularly after the esterification reaction thus becoming an option for the formulation of new antioxidant products. PMID:25104002

  2. Effects of cerium oxide nanoparticles on soil enzymatic activities and wheat grass nutrients uptake

    Science.gov (United States)

    Li, Biting; Chen, Yirui; Bai, Lingyun; Jacobson, Astrid; Darnault, Christophe

    2015-04-01

    The US National Science Foundation estimated that the use of nanomaterials and nanotechnology would reach a global market value of 1 million this year. Concomitant with the wide applications of nanoparticles is an increasing risk of adverse effects to the environment and human health. As a common nanomaterial used as a fuel catalyst and polish material, cerium (IV) oxide nanoparticles (CeO2 NP) were tested for their potential impact on soil health and plant growth. Through exposure by air, water, and solid deposition, nanoparticles may accumulate in soils and impact agricultural systems. The objectives of this research were to determine whether CeO2 NPs affect the growth of wheat grass and selected soil enzyme activities chose as indicators of soil health. Wheat grass was grown in plant boxes containing CeO2 NPs mixed with agricultural soil at different concentrations. Two control groups were included: one consisting of soil with plants but no CeO2 NPs, and one containing only soil, i.e., no NP or wheat plants added. The plants were grown for 10 weeks and harvested every two weeks in a laboratory under sodium growth lights. At the end of the each growing period, two weeks, soils were assayed for phosphatase, β-glucosidase, and urease activities, and NPK values. Spectrophotometer analyses were used to assess enzyme activities, and NPK values were tested by Clemson Agricultural Center. Wheat yields were estimated by shoot and root lengths and weights.

  3. Screening for Genes Coding for Putative Antitumor Compounds, Antimicrobial and Enzymatic Activities from Haloalkalitolerant and Haloalkaliphilic Bacteria Strains of Algerian Sahara Soils

    Directory of Open Access Journals (Sweden)

    Okba Selama

    2014-01-01

    Full Text Available Extreme environments may often contain unusual bacterial groups whose physiology is distinct from those of normal environments. To satisfy the need for new bioactive pharmaceuticals compounds and enzymes, we report here the isolation of novel bacteria from an extreme environment. Thirteen selected haloalkalitolerant and haloalkaliphilic bacteria were isolated from Algerian Sahara Desert soils. These isolates were screened for the presence of genes coding for putative antitumor compounds using PCR based methods. Enzymatic, antibacterial, and antifungal activities were determined by using cultural dependant methods. Several of these isolates are typical of desert and alkaline saline soils, but, in addition, we report for the first time the presence of a potential new member of the genus Nocardia with particular activity against the yeast Saccharomyces cerevisiae. In addition to their haloalkali character, the presence of genes coding for putative antitumor compounds, combined with the antimicrobial activity against a broad range of indicator strains and their enzymatic potential, makes them suitable for biotechnology applications.

  4. Covalent immobilization of porcine pancreatic lipase on carboxyl-activated magnetic nanoparticles: characterization and application for enzymatic inhibition assays.

    Science.gov (United States)

    Zhu, Yuan-Ting; Ren, Xiao-Yun; Liu, Yi-Ming; Wei, Ying; Qing, Lin-Sen; Liao, Xun

    2014-05-01

    Using carboxyl functionalized silica-coated magnetic nanoparticles (MNPs) as carrier, a novel immobilized porcine pancreatic lipase (PPL) was prepared through the 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. Transmission electron microscopic images showed that the synthesized nanoparticles (Fe3O4-SiO2) possessed three dimensional core-shell structures with an average diameter of ~20 nm. The effective enzyme immobilization onto the nanocomposite was confirmed by atomic force microscopic (AFM) analysis. Results from Fourier-transform infrared spectroscopy (FT-IR), Bradford protein assay, and thermo-gravimetric analysis (TGA) indicated that PPL was covalently attached to the surface of magnetic nanoparticles with a PPL immobilization yield of 50mg enzyme/g MNPs. Vibrating sample magnetometer (VSM) analysis revealed that the MNPs-PPL nanocomposite had a high saturation magnetization of 42.25 emu·g(-1). The properties of the immobilized PPL were investigated in comparison with the free enzyme counterpart. Enzymatic activity, reusability, thermo-stability, and storage stability of the immobilized PPL were found significantly superior to those of the free one. The Km and the Vmax values (0.02 mM, 6.40 U·mg(-1) enzyme) indicated the enhanced activity of the immobilized PPL compared to those of the free enzyme (0.29 mM, 3.16 U·mg(-1) enzyme). Furthermore, at an elevated temperature of 70 °C, immobilized PPL retained 60% of its initial activity. The PPL-MNPs nanocomposite was applied in the enzyme inhibition assays using orlistat, and two natural products isolated from oolong tea (i.e., EGCG and EGC) as the test compounds. PMID:24656379

  5. Research on the qualitative enzymatic activities in sediments from Secu and Gozna-Văliug dam reservoirs, Caraş-Severin county (Romania

    Directory of Open Access Journals (Sweden)

    Mihail Drăgan-Bularda

    2011-12-01

    Full Text Available Six sediment samples (three from Secu Lake and three from Gozna-Văliug Lake werecollected and analyzed qualitatively, enzymologically. In the sediment samples, the followingenzymatic activities have been qualitatively determined: four oligase activities (maltase, saccharase,lactase and cellobiase and five polyase activities (amylase, cellulose, dextranase, glicogenase andinulinase. The studied activities were determined in each sample and they were found to displayvariations in the intensities of the processes depending on the sampling place. Generally, the highestintensity of qualitative enzymatic activities was registered in case of oligases.

  6. Phospholipase C Produced by Clostridium botulinum Types C and D:Comparison of Gene, Enzymatic, and Biological Activities with Those of Clostridium perfringens Alpha-toxin

    Directory of Open Access Journals (Sweden)

    Sakurai,Jun

    2013-02-01

    Full Text Available Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs, the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin. Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity;however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival.

  7. Enzymatic activity of fungi of the genus Candida isolated from the skin and the digestive tract in people and from municipal sewage

    Directory of Open Access Journals (Sweden)

    Maria Dynowska

    2014-08-01

    Full Text Available Enzymatic activities of C. albicans, C. guilliermondii and C. tropicalis from the skin and the digestive tract and from municipal sewage were compared using the API ZYM system (bio Méricux. Aclivities were examined in relalion to potential pathogenicity of these fungi strictly connected with their enzymatic properties, especilly the production of proteolytic and lipolytic enzymes, as well as acidic and basic phosphatase. The proteolytic activity was high in strains of C. albicans from the digestive tract and from municipal sewage, as we~ as in C. guilliermondii isolated from sewage. The highest lipolytic activity was recorded in the case of C. guilliermondii from sewage, and C. albicans and C. tropicalis from the digestive tract. Acidic and basic phosphatases and phosphohydrolase were secreted by strains of all species isolated from municipal sewage in the greatest degree (the only exception is a high activity of acidic phosphatase of C. albicans and C. tropicalis in the digestive tract. A higher enzymatic activity was observed in the same species in municipal sewage than in the human body.

  8. E. coli-Derived L-Asparaginase Retains Enzymatic and Cytotoxic Activity In Vitro for Canine and Feline Lymphoma after Cold Storage

    Directory of Open Access Journals (Sweden)

    Jackie M. Wypij

    2013-01-01

    Full Text Available Background. L-asparaginase is effective in treating canine and feline lymphoma, however chemotherapy poses a significant financial cost to veterinary clients, limiting therapy for many pets. Single dose vials result in significant drug wastage, and drug shortages limit consistent availability for pets. Hypothesis. E. coli-derived asparaginase retains enzymatic and antineoplastic activity in canine and feline lymphoma cells after cold storage. Methods. E. coli-derived asparaginase was cold-stored: refrigeration (7–14 days and freezing (14 days–six months, one to three freeze/thaw cycles. Enzymatic activity of asparaginase was measured via a modified asparagine assay. Effects of cold-stored asparaginase on cell proliferation and cytotoxicity were measured in feline (MYA-1, F1B and canine (17–71, OSW lymphoma cells. Results. Cold-stored E. coli-derived asparaginase retains antineoplastic activity in all four cell lines tested. Cold-stored E. coli-derived L-asparaginase depletes asparagine and retains enzymatic activity. Duration of refrigeration, duration of freezing, and number of freeze-thaw cycles have minimal effect on asparaginase enzyme activity. Conclusions and Clinical Importance. This study establishes a scientific basis for long-term cold storage of reconstituted E. coli-derived asparaginase that may result in better utilization of limited drug resources and improve financial feasibility of E. coli-derived asparaginase as a therapeutic option for pets with lymphoma.

  9. Effect of alcoholic extracts of Indian medicinal plants on the altered enzymatic activities of diabetic rats

    Directory of Open Access Journals (Sweden)

    Sundaram E

    2009-01-01

    Full Text Available In present study, the effect of alcoholic extract of Momordica charantia, Aegle marmelos and Eugenia jambolana was studied on serum glutamic oxaloacetate transminase and serum glutamic pyruvate transminase activities and on serum urea, total protein and albumin concentrations of streptozotocin diabetic rats. Diabetes in rats was induced by single dose of streptozotocin (30 mg/kg i. p.. On confirming the diabetes after 48 h of injection, alcoholic extracts of three plants were administered orally in doses of 250 mg and 500 mg/kg/d for 30 d. Glibenclamide (300 µg/kg/d was used as a reference drug for comparison. Streptozotocin diabetic rats showed a significant increase in serum glutamic oxaloacetate transminase and serum glutamic pyruvate transminase activities and serum urea concentration but a significant decrease in serum total protein and albumin concentrations and albumin/globulin ratio. Oral administration of alcoholic extract of Momordica charantia, Aegle marmelos and Eugenia jambolana in daily doses of 250 mg and 500 mg/kg for a period of 1 mo produced dose- and duration-dependent decrease in serum glutamic oxaloacetate transminase and serum glutamic pyruvate transminase activities as well as decrease in serum urea concentration and restored the serum total protein and albumin concentration and albumin/globulin ratio to a great extent in streptozotocin diabetic rats. The beneficial effects of these plants in 500 mg/kg dose in streptozotocin diabetic rats were comparable to that of glibenclamide (300 µg/kg, a standard oral hypoglycaemic drug used in clinical practice.

  10. Environmental Factors Modulating the Stability and Enzymatic Activity of the Petrotoga mobilis Esterase (PmEst.

    Directory of Open Access Journals (Sweden)

    Jose L S Lopes

    Full Text Available Enzymes isolated from thermophilic organisms found in oil reservoirs can find applications in many fields, including the oleochemical, pharmaceutical, bioenergy, and food/dairy industries. In this study, in silico identification and recombinant production of an esterase from the extremophile bacteria Petrotoga mobilis (designated PmEst were performed. Then biochemical, bioinformatics and structural characterizations were undertaken using a combination of synchrotron radiation circular dichroism (SRCD and fluorescence spectroscopies to correlate PmEst stability and hydrolytic activity on different substrates. The enzyme presented a high Michaelis-Menten constant (KM 0.16 mM and optimum activity at ~55°C for p-nitrophenyl butyrate. The secondary structure of PmEst was preserved at acid pH, but not under alkaline conditions. PmEst was unfolded at high concentrations of urea or guanidine through apparently different mechanisms. The esterase activity of PmEst was preserved in the presence of ethanol or propanol and its melting temperature increased ~8°C in the presence of these organic solvents. PmEst is a mesophilic esterase with substrate preference towards short-to medium-length acyl chains. The SRCD data of PmEst is in agreement with the prediction of an α/β protein, which leads us to assume that it displays a typical fold of esterases from this family. The increased enzyme stability in organic solvents may enable novel applications for its use in synthetic biology. Taken together, our results demonstrate features of the PmEst enzyme that indicate it may be suitable for applications in industrial processes, particularly, when the use of polar organic solvents is required.

  11. Enhancing phytochemical levels, enzymatic and antioxidant activity of spinach leaves by chitosan treatment and an insight into the metabolic pathway using DART-MS technique.

    Science.gov (United States)

    Singh, Shachi

    2016-05-15

    Phytochemicals are health promoting compounds, synthesized by the plants to protect them against biotic or abiotic stress. The metabolic pathways leading to the synthesis of these phytochemicals are highly inducible; therefore methods could be developed to enhance their production by the exogenous application of chemical inducers/elicitors. In the present experiment, chitosan was used as an elicitor molecule to improve the phytochemical content of spinach plant. When applied at a concentration of 0.01 mg/ml as a foliar spray, chitosan was able to cause an increase in the enzymatic (peroxidase, catalase and phenylalanine ammonium lyase (PAL)) and non enzymatic (total phenolics, flavonoids and proteins) defensive metabolites, as well as, in the total antioxidant activity of the spinach leaves. A 1.7-fold increase in the total phenolics, a 2-fold increase in total flavonoid and a 1.6-fold increase in total protein were achieved with the treatment. A higher level of enzymatic activity was observed with a 4-fold increase in peroxidase and approximately 3-fold increases in catalase and phenylalanine ammonium lyase activity. Antioxidant activity showed a positive correlation between phenolic compounds and the enzymatic activity. Direct analysis in real time mass spectrometry (DART-MS) was applied to generate the metabolite profile of control and treated leaves. DART analysis revealed the activation of phenylpropanoid pathway by chitosan molecule, targeting the synthesis of diverse classes of flavonoids and their glycosides. Important metabolites of stress response were also visible in the DART spectra, including proline and free sugars. PMID:26775959

  12. HISTOPATHOLOGICAL CHANGES AND ENZYMATIC ACTIVITIES INDUCED BY MELOIDOGYNE INCOGNITA ON RESISTANT AND SUSCEPTIBLE POTATO

    Directory of Open Access Journals (Sweden)

    Moawad M. Mohamad

    2012-12-01

    Full Text Available All potato cultivars are susceptible to root-knot nematodes (Meloidogyne spp. which infest the roots and induce galls on the surface and necrotic spots in the flesh tuber of potato, Solanum tuberosum. Infested tubers are unacceptable for processing and fresh market. Tubers are also putative source of dissemination of the nematode. A French nematode- resistant tetraploid potato genotype gained from ex-S. sparsipilum material hybridized with S. tuberosum in F1 and in their back cross progenies and designated as 02T.155.6 was tested and compared in the present study in Egypt as a suitable different environment. Histopathological changes and chitinase activity induced by M. incognita population, of common occurrence in Egypt, in four French tetraploid materials and two common cultivars known as nematode- resistant and susceptible potato genotypes were investigated. Hypertrophied cells were initiated in both cortical and steler regions of the roots which were then developed to abnormal xylem elements expanding into the cortex in French susceptible genotypes designated as 02T.149.6, 02T.150.54, and 02T.157.16. Nematode within the vascular tissue (stele could induce giant cell development close to nematode heads. The largest number of such induced cells was shown by the cultivars Spunta and Diamant. The clone 02T.155.6 with putative nematode resistance demonstrated none or very little nematode development. Recently dead second stage juveniles could also indicate incompatible plant reaction to the invading nematodes in 02T.155.6. M. incognita, Giza population, resistance was generally more coherent to 02T.155.6 as demonstrated by our histological investigations but less coherent as shown by another Egyptian M. incognita population. Chitinase activity was enhanced in M. incognita (Giza-inoculated with respect to uninoculated roots in all plants. After inoculation, such an activity generally increased more in roots of a potato genotype previously known to

  13. An enzymatic activity isolated from Brassica oleracea specific for UV-irradiated DNA

    International Nuclear Information System (INIS)

    As a consequence of a breakdown in the ozone layer, an increase in the amount of DNA damage caused by ultraviolet irradiation can be expected. Organisms have evolved mechanisms to repair numerous types of DNA damages. While these DNA repair systems have been well characterized in bacteria and to a lesser extent in mammalian cells, surprisingly little is known about repair of potentially harmful DNA lesions in plants. An enzyme that recognizes and incises UV irradiated DNA has been partially purified from the leaf tissue of Brassica oleracea. Glycosylase-produced base loss sites were detected by a nitrocellulose filter-binding assay using UV-irradiated PM2 viral DNA as the substrate. The optimal temperature for maximal enzyme activity is 47C with a pH optimum between 7.0 and 7.5. In addition, the endonuclease is active in both Tris and phosphate buffers, although it is stimulated by phosphate concentrations up to 25 mM. Currently, a number of synthetic polynucleotides as well as DNAs of defined sequence are being employed as substrates to determine the nature of the UV-induced lesion and the precise mechanism of action of the enzyme

  14. Lignin binding to pancreatic lipase and its influence on enzymatic activity.

    Science.gov (United States)

    Zhang, Juan; Xiao, Lin; Yang, Yucai; Wang, Zhaoxia; Li, Genxi

    2014-04-15

    In this paper, we find that the effect of lignin on pancreatic lipase (PL) is dependent on reaction medium and substrate used. Experimental results reveal that lignin can gradually bind to PL to form a PL-lignin complex, resulting in an increased activity of the enzyme. The binding process is spontaneous and the PL-lignin complex formation is an endothermic reaction induced by hydrophobic and electrostatic interaction. There is a non-radiation energy transfer from PL to lignin during the binding process, and the binding of lignin to PL conforms to a secondary exponential decay function. Moreover, the α-helix content of the enzyme will be changed and the rigidity of its side chain will be enhanced due to the formation of lignin-PL complex. This study has not only provided the activation effect of lignin on PL, but also given an insight into the interaction between lignin and the enzyme, which would benefit the application of lignin in the pharmacy and food industry, as well as other fields. PMID:24295682

  15. Characterization of 10-hydroxygeraniol dehydrogenase from Catharanthus roseus reveals cascaded enzymatic activity in iridoid biosynthesis.

    Science.gov (United States)

    Krithika, Ramakrishnan; Srivastava, Prabhakar Lal; Rani, Bajaj; Kolet, Swati P; Chopade, Manojkumar; Soniya, Mantri; Thulasiram, Hirekodathakallu V

    2015-01-01

    Catharanthus roseus [L.] is a major source of the monoterpene indole alkaloids (MIAs), which are of significant interest due to their therapeutic value. These molecules are formed through an intermediate, cis-trans-nepetalactol, a cyclized product of 10-oxogeranial. One of the key enzymes involved in the biosynthesis of MIAs is an NAD(P)(+) dependent oxidoreductase system, 10-hydroxygeraniol dehydrogenase (Cr10HGO), which catalyses the formation of 10-oxogeranial from 10-hydroxygeraniol via 10-oxogeraniol or 10-hydroxygeranial. This work describes the cloning and functional characterization of Cr10HGO from C. roseus and its role in the iridoid biosynthesis. Substrate specificity studies indicated that, Cr10HGO has good activity on substrates such as 10-hydroxygeraniol, 10-oxogeraniol or 10-hydroxygeranial over monohydroxy linear terpene derivatives. Further it was observed that incubation of 10-hydroxygeraniol with Cr10HGO and iridoid synthase (CrIDS) in the presence of NADP(+) yielded a major metabolite, which was characterized as (1R, 4aS, 7S, 7aR)-nepetalactol by comparing its retention time, mass fragmentation pattern, and co-injection studies with that of the synthesized compound. These results indicate that there is concerted activity of Cr10HGO with iridoid synthase in the formation of (1R, 4aS, 7S, 7aR)-nepetalactol, an important intermediate in iridoid biosynthesis. PMID:25651761

  16. Evolution of Enzymatic Activities in the Enolase Superfamily: L-Fuconate Dehydratase from Xanthomonas campestris

    Energy Technology Data Exchange (ETDEWEB)

    Yew,W.; Fedorov, A.; Fedorov, E.; Rakus, J.; Pierce, R.; Almo, S.; Gerlt, J.

    2006-01-01

    Many members of the mechanistically diverse enolase superfamily have unknown functions. In this report the authors use both genome (operon) context and screening of a library of acid sugars to assign the L-fuconate dehydratase (FucD) function to a member of the mandelate racemase (MR) subgroup of the superfamily encoded by the Xanthomonas campestris pv. campestris str. ATCC 33913 genome (GI: 21233491). Orthologues of FucD are found in both bacteria and eukaryotes, the latter including the rTS beta protein in Homo sapiens that has been implicated in regulating thymidylate synthase activity. As suggested by sequence alignments and confirmed by high-resolution structures in the presence of active site ligands, FucD and MR share the same active site motif of functional groups: three carboxylate ligands for the essential Mg2+ located at the ends of th third, fourth, and fifth-strands in the (/)7-barrel domain (Asp 248, Glu 274, and Glu 301, respectively), a Lys-x-Lys motif at the end of the second-strand (Lys 218 and Lys 220), a His-Asp dyad at the end of the seventh and sixth-strands (His 351 and Asp 324, respectively), and a Glue at the end of the eighth-strand (Glu 382). The mechanism of the FucD reaction involves initial abstraction of the 2-proton by Lys 220, acid catalysis of the vinylogous-elimination of the 3-OH group by His 351, and stereospecific ketonization of the resulting 2-keto-3-deoxy-L-fuconate product. Screening of the library of acid sugars revealed substrate and functional promiscuity: In addition to L-fuconate, FucD also catalyzes the dehydration of L-galactonate, D-arabinonate, D-altronate, L-talonate, and D-ribonate. The dehydrations of L-fuconate, L-galactonate, and D-arabinonate are initiated by abstraction of the 2-protons by Lys 220. The dehydrations of L-talonate and D-ribonate are initiated by abstraction of the 2-protons by His 351; however, protonation of the enediolate intermediates by the conjugate acid of Lys 220 yields L

  17. The antihyperlipidemic activities of enzymatic and acidic intracellular polysaccharides by Termitomyces albuminosus.

    Science.gov (United States)

    Zhao, Huajie; Li, Shangshang; Zhang, Jianjun; Che, Gen; Zhou, Meng; Liu, Min; Zhang, Chen; Xu, Nuo; Lin, Lin; Liu, Yu; Jia, Le

    2016-10-20

    Two polysaccharides, EIPS and AIPS were obtained by the hydrolysis of IPS from Termitomyces albuminosus, and their pharmacological effects on blood lipid profiles metabolism and oxidative stress were investigated. The results demonstrated that EIPS was superior to IPS and AIPS on reducing hepatic lipid levels and preventing oxidative stress by improving serum enzyme activities (ALT, AST, and ALP), serum lipid levels (TC, TG, HDL-C, LDL-C and VLDL-C), hepatic lipid levels (TC and TG), and antioxidant status (SOD, GSH-Px, CAT, T-AOC, MDA, and LPO). These conclusions indicated that EIPS, AIPS and IPS might be suitable for functional foods and natural drugs on preventing the high-fat emulsion-induced hyperlipidemia. In addition, the monosaccharide compositions of IPS and its hydrolyzate were also processed. PMID:27474674

  18. Antioxidant activities of buttermilk proteins, whey proteins, and their enzymatic hydrolysates.

    Science.gov (United States)

    Conway, Valérie; Gauthier, Sylvie F; Pouliot, Yves

    2013-01-16

    The oxygen radical absorbance capacities (ORAC) and metal chelating capacities (MCC) of protein concentrates prepared from buttermilk and cheese whey by ultrafiltration were compared with those of skim milk protein. Samples were also heat-denatured and hydrolyzed by pepsin for 2 h followed by trypsin for 3 h. The highest MCC was obtained for hydrolyzed skim milk protein. ORAC values ranged from 554.4 to 1319.6 μmol Trolox equivalents/g protein, with the highest value obtained for hydrolyzed buttermilk protein. Liquid-phase isoelectric focusing (IEF) of this hydrolysate yielded peptide fractions with lower ORAC values. LC-MS analysis of the hydrolyzed skim milk and buttermilk proteins and IEF fractions of the latter showed that peptides derived from milk fat globule membrane proteins, primarily butyrophilin, could be responsible for the superior antioxidant activity of buttermilk. These results suggest overall that hydrolyzed buttermilk protein could be used as a source of natural antioxidants. PMID:23244578

  19. Factors influencing the rate of non-enzymatic activation of carboxylic and amino acids by ATP

    Science.gov (United States)

    Mullins, D. W., Jr.; Lacey, J. C., Jr.

    1981-01-01

    The nonenzymatic formation of adenylate anhydrides of carboxylic and amino acids is discussed as a necessary step in the origin of the genetic code and protein biosynthesis. Results of studies are presented which have shown the rate of activation to depend on the pKa of the carboxyl group, the pH of the medium, temperature, the divalent metal ion catalyst, salt concentration, and the nature of the amino acid. In particular, it was found that of the various amino acids investigated, phenylalanine had the greatest affinity for the adenine derivatives adenosine and ATP. Results thus indicate that selective affinities between amino acids and nucleotides were important during prebiotic chemical evolution, and may have played a major role in the origin of protein synthesis and genetic coding.

  20. Rapid Bioorthogonal Chemistry Turn-on through Enzymatic or Long Wavelength Photocatalytic Activation of Tetrazine Ligation.

    Science.gov (United States)

    Zhang, Han; Trout, William S; Liu, Shuang; Andrade, Gabriel A; Hudson, Devin A; Scinto, Samuel L; Dicker, Kevin T; Li, Yi; Lazouski, Nikifar; Rosenthal, Joel; Thorpe, Colin; Jia, Xinqiao; Fox, Joseph M

    2016-05-11

    Rapid bioorthogonal reactivity can be induced by controllable, catalytic stimuli using air as the oxidant. Methylene blue (4 μM) irradiated with red light (660 nm) catalyzes the rapid oxidation of a dihydrotetrazine to a tetrazine thereby turning on reactivity toward trans-cyclooctene dienophiles. Alternately, the aerial oxidation of dihydrotetrazines can be efficiently catalyzed by nanomolar levels of horseradish peroxidase under peroxide-free conditions. Selection of dihydrotetrazine/tetrazine pairs of sufficient kinetic stability in aerobic aqueous solutions is key to the success of these approaches. In this work, polymer fibers carrying latent dihydrotetrazines were catalytically activated and covalently modified by trans-cyclooctene conjugates of small molecules, peptides, and proteins. In addition to visualization with fluorophores, fibers conjugated to a cell adhesive peptide exhibited a dramatically increased ability to mediate contact guidance of cells. PMID:27078610

  1. Dimerization and enzymatic activity of fungal 17β-hydroxysteroid dehydrogenase from the short-chain dehydrogenase/reductase superfamily

    Directory of Open Access Journals (Sweden)

    Kristan Katja

    2005-12-01

    Full Text Available Abstract Background 17β-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17β-HSDcl is a member of the short-chain dehydrogenase/reductase (SDR superfamily. SDR proteins usually function as dimers or tetramers and 17β-HSDcl is also a homodimer under native conditions. Results We have investigated here which secondary structure elements are involved in the dimerization of 17β-HSDcl and examined the importance of dimerization for the enzyme activity. Sequence similarity with trihydroxynaphthalene reductase from Magnaporthe grisea indicated that Arg129 and His111 from the αE-helices interact with the Asp121, Glu117 and Asp187 residues from the αE and αF-helices of the neighbouring subunit. The Arg129Asp and His111Leu mutations both rendered 17β-HSDcl monomeric, while the mutant 17β-HSDcl-His111Ala was dimeric. Circular dichroism spectroscopy analysis confirmed the conservation of the secondary structure in both monomers. The three mutant proteins all bound coenzyme, as shown by fluorescence quenching in the presence of NADP+, but both monomers showed no enzymatic activity. Conclusion We have shown by site-directed mutagenesis and structure/function analysis that 17β-HSDcl dimerization involves the αE and αF helices of both subunits. Neighbouring subunits are connected through hydrophobic interactions, H-bonds and salt bridges involving amino acid residues His111 and Arg129. Since the substitutions of these two amino acid residues lead to inactive monomers with conserved secondary structure, we suggest dimerization is a prerequisite for catalysis. A detailed understanding of this dimerization could lead to the development of compounds that will specifically prevent dimerization, thereby serving as a new type of inhibitor.

  2. Sulfated polysaccharide extracted of the green algae Caulerpa racemosa increase the enzymatic activity and paw edema induced by sPLA2 from Crotalus durissus terrificus venom

    Directory of Open Access Journals (Sweden)

    Camila L. Pires

    2013-08-01

    Full Text Available Sulfated polysaccharides derived from seaweed have shown great potential for use in the development of new drugs. In this study, we observed that a low-molecular-weight sulfated polysaccharide from Caulerpa racemosa, termed CrSP, could interact with secretory phospholipase A2 (sPLA2 isolated from Crotalus durissus terrificus venom. When native sPLA2 (14 kDa was incubated with CrSP, they formed a molecular complex (sPLA2:CrSP with a molecular mass of 32 kDa, approximately. Size exclusion chromatography experiments suggested that CrSP formed a stable complex with sPLA2. We belived that sPLA2 and SPCr are involved an ionic interaction between negatively charged CrSP and the positively charged basic amino acid residues of sPLA2, because this interaction induced significant changes in sPLA2 enzymatic and pharmacological activities. CrSP caused a significant increase in sPLA2 enzymatic and bactericidal activity and increased its edematogenic effect. A pharmacological assay showed that the myotoxic activity of sPLA2:CrSP is unrelated to its enzymatic activity and that sPLA2:CrSP may have a practical application as a natural antibacterial agent for use in humans and commercially raised animals.

  3. Enzymatic studies on phosphorus availability from phosphate compounds by microbial activities using nuclear techniques

    International Nuclear Information System (INIS)

    the present study aimed to evaluate to evaluate the of phosphate solubilizing bacteria (PSB) and vesicular arbuscular mycorrhizae(VAM) as microbiological mean to convert the sparingly soluble P into available from utilized by plant through excretion of acid and alkaline phosphatase. rock phosphate as natural and a cheap source of P- fertilizer was applied in the present study. To trace the effect of microbial activity and phosphatase enzymes on phosphate availability from its compounds , set of experiments were conducted either in the lab. or green house. The obtained results could be summarized as following:- 1- phosphate solubilizing bacteria(PSB) w isolated from samples of fertile soil, 16 colonies showed positive reaction were chosen .2- bacteria , which exhibited high phosphate clearing zone (PCZ) selected to detect their efficiencies for dissolving rock phospate and select the effective one (most potent) on the basis of highest production of phosphatase and phosphorus solubilization.3- identification of the most potent (pseudomonas aeruginosa) 4- effective environmental and nutritional factors on phosphatase production by pseudomonas aeruginosa were discussed.green-house experiment: inoculation of wheat (triticum aestivum cv.sakha 8) with either PSB and /or VAM with or without rock-P fertilization was under taken. dual inoculation with psb (pseudomonas aeruginosa) and VAM improved the dry matter yield and N and P uptake by wheat as compared to other treatments. application of psb as well as VAM increased the availability of rock phosphate to be utilized by wheat

  4. Toward an Automatic Determination of Enzymatic Reaction Mechanisms and Their Activation Free Energies.

    Science.gov (United States)

    Zinovjev, Kirill; Ruiz-Pernía, J Javier; Tuñón, Iñaki

    2013-08-13

    We present a combination of the string method and a path collective variable for the exploration of the free energy surface associated to a chemical reaction in condensed environments. The on-the-fly string method is employed to find the minimum free energy paths on a multidimensional free energy surface defined in terms of interatomic distances, which is a convenient selection to study bond forming/breaking processes. Once the paths have been determined, a reaction coordinate is defined as a measure of the advance of the system along these paths. This reaction coordinate can be then used to trace the reaction Potential of Mean Force from which the activation free energy can be obtained. This combination of methodologies has been here applied to the study, by means of Quantum Mechanics/Molecular Mechanics simulations, of the reaction catalyzed by guanidinoacetate methyltransferase. This enzyme catalyzes the methylation of guanidinoacetate by S-adenosyl-l-methionine, a reaction that involves a methyl transfer and a proton transfer and for which different reaction mechanisms have been proposed. PMID:26584125

  5. Computational modeling of the enzymatic activities of biomolecules at different scales: from quantum mechanical reaction studies to systemic understanding of cell behavior

    OpenAIRE

    Barbieri,, R.

    2012-01-01

    The aim of the thesis is the development of computational models of the enzymatic activity of biomolecules at different scales. Parallel investigations have been carried out at a quantum level to study the reactivity of an enzyme from an electronic point of view, and at a systemic level using simulation techniques to determine the role of enzymes in the network of cellular reactions. Starting from the lowest complexity level, the thesis begins with two computational studies with the aims ...

  6. Role of trypsin-like cleavage at arginine 192 in the enzymatic and cytotonic activities of Escherichia coli heat-labile enterotoxin.

    OpenAIRE

    C.C. Grant; Messer, R J; Cieplak, W

    1994-01-01

    Previous studies of cholera toxin and Escherichia coli heat-labile enterotoxin have suggested that proteolytic cleavage plays an important role in the expression of ADP-ribosyltransferase activity and toxicity. Specifically, several studies have implicated a trypsin-like cleavage at arginine 192, which lies within an exposed region subtended by a disulfide bond in the intact A subunit, in toxicity. To investigate the role of this modification in the enzymatic and cytotonic properties of heat-...

  7. Direct electrocatalytic reduction of coenzyme NAD{sup +} to enzymatically-active 1,4-NADH employing an iridium/ruthenium-oxide electrode

    Energy Technology Data Exchange (ETDEWEB)

    Ullah, Nehar, E-mail: nehar.ullah@mail.mcgill.ca; Ali, Irshad; Omanovic, Sasha

    2015-01-15

    A thermally prepared iridium/ruthenium-oxide coating (Ir{sub 0.8}Ru{sub 0.2}-oxide) formed on a titanium substrate was investigated as a possible electrode for direct electrochemical regeneration of enzymatically-active 1,4-NADH from its oxidized form NAD{sup +}, at various electrode potentials, in a batch electrochemical reactor. The coating surface was characterized by ‘cracked mud’ morphology, yielding a high surface roughness. The NADH regeneration results showed that the percentage of enzymatically-active 1,4-NADH present in the product mixture (i.e. recovery) is strongly dependent on the electrode potential, reaching a maximum (88%) at −1.70 V vs. MSE. The relatively high recovery was explained on the basis of availability of adsorbed ‘active’ hydrogen (H{sub ads}) on the Ir/Ru-oxide surface, i.e. on the basis of electrochemical hydrogenation. - Highlights: • Ir{sub 0.8}Ru{sub 0.2}-oxide coating was formed thermally on a Ti substrate. • Electrochemical regeneration of enzymatically-active 1,4-NADH was investigated. • The 1,4-NADH recovery percentage is strongly dependent on the electrode potential. • A highest recovery, 88%, was obtained at −1.70 V vs. MSE. • The NADH regeneration process involved electrochemical hydrogenation.

  8. Direct electrocatalytic reduction of coenzyme NAD+ to enzymatically-active 1,4-NADH employing an iridium/ruthenium-oxide electrode

    International Nuclear Information System (INIS)

    A thermally prepared iridium/ruthenium-oxide coating (Ir0.8Ru0.2-oxide) formed on a titanium substrate was investigated as a possible electrode for direct electrochemical regeneration of enzymatically-active 1,4-NADH from its oxidized form NAD+, at various electrode potentials, in a batch electrochemical reactor. The coating surface was characterized by ‘cracked mud’ morphology, yielding a high surface roughness. The NADH regeneration results showed that the percentage of enzymatically-active 1,4-NADH present in the product mixture (i.e. recovery) is strongly dependent on the electrode potential, reaching a maximum (88%) at −1.70 V vs. MSE. The relatively high recovery was explained on the basis of availability of adsorbed ‘active’ hydrogen (Hads) on the Ir/Ru-oxide surface, i.e. on the basis of electrochemical hydrogenation. - Highlights: • Ir0.8Ru0.2-oxide coating was formed thermally on a Ti substrate. • Electrochemical regeneration of enzymatically-active 1,4-NADH was investigated. • The 1,4-NADH recovery percentage is strongly dependent on the electrode potential. • A highest recovery, 88%, was obtained at −1.70 V vs. MSE. • The NADH regeneration process involved electrochemical hydrogenation

  9. Covalent immobilization of porcine pancreatic lipase on carboxyl-activated magnetic nanoparticles: Characterization and application for enzymatic inhibition assays

    International Nuclear Information System (INIS)

    Using carboxyl functionalized silica-coated magnetic nanoparticles (MNPs) as carrier, a novel immobilized porcine pancreatic lipase (PPL) was prepared through the 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. Transmission electron microscopic images showed that the synthesized nanoparticles (Fe3O4–SiO2) possessed three dimensional core–shell structures with an average diameter of ∼ 20 nm. The effective enzyme immobilization onto the nanocomposite was confirmed by atomic force microscopic (AFM) analysis. Results from Fourier-transform infrared spectroscopy (FT-IR), Bradford protein assay, and thermo-gravimetric analysis (TGA) indicated that PPL was covalently attached to the surface of magnetic nanoparticles with a PPL immobilization yield of 50 mg enzyme/g MNPs. Vibrating sample magnetometer (VSM) analysis revealed that the MNPs-PPL nanocomposite had a high saturation magnetization of 42.25 emu·g−1. The properties of the immobilized PPL were investigated in comparison with the free enzyme counterpart. Enzymatic activity, reusability, thermo-stability, and storage stability of the immobilized PPL were found significantly superior to those of the free one. The Km and the Vmax values (0.02 mM, 6.40 U·mg−1 enzyme) indicated the enhanced activity of the immobilized PPL compared to those of the free enzyme (0.29 mM, 3.16 U·mg−1 enzyme). Furthermore, at an elevated temperature of 70 °C, immobilized PPL retained 60% of its initial activity. The PPL-MNPs nanocomposite was applied in the enzyme inhibition assays using orlistat, and two natural products isolated from oolong tea (i.e., EGCG and EGC) as the test compounds. - Highlights: • Porcine pancreatic lipase was firstly covalently immobilized onto carboxylated MNPs. • Immobilized porcine pancreatic lipase (PPL) was characterized by various techniques. • MNPs-PPL showed higher activity, reusability, and thermo-stability than the free PPL

  10. Covalent immobilization of porcine pancreatic lipase on carboxyl-activated magnetic nanoparticles: Characterization and application for enzymatic inhibition assays

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Yuan-Ting [Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Ren, Xiao-Yun [Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041 (China); Liu, Yi-Ming [Department of Chemistry and Biochemistry, Jackson State University, 1400 Lynch St., Jackson, MS 39217 (United States); Wei, Ying [Changzhi Medical College, Changzhi 046000 (China); Qing, Lin-Sen [Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041 (China); Liao, Xun, E-mail: liaoxun@cib.ac.cn [Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041 (China)

    2014-05-01

    Using carboxyl functionalized silica-coated magnetic nanoparticles (MNPs) as carrier, a novel immobilized porcine pancreatic lipase (PPL) was prepared through the 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. Transmission electron microscopic images showed that the synthesized nanoparticles (Fe{sub 3}O{sub 4}–SiO{sub 2}) possessed three dimensional core–shell structures with an average diameter of ∼ 20 nm. The effective enzyme immobilization onto the nanocomposite was confirmed by atomic force microscopic (AFM) analysis. Results from Fourier-transform infrared spectroscopy (FT-IR), Bradford protein assay, and thermo-gravimetric analysis (TGA) indicated that PPL was covalently attached to the surface of magnetic nanoparticles with a PPL immobilization yield of 50 mg enzyme/g MNPs. Vibrating sample magnetometer (VSM) analysis revealed that the MNPs-PPL nanocomposite had a high saturation magnetization of 42.25 emu·g{sup −1}. The properties of the immobilized PPL were investigated in comparison with the free enzyme counterpart. Enzymatic activity, reusability, thermo-stability, and storage stability of the immobilized PPL were found significantly superior to those of the free one. The K{sub m} and the V{sub max} values (0.02 mM, 6.40 U·mg{sup −1} enzyme) indicated the enhanced activity of the immobilized PPL compared to those of the free enzyme (0.29 mM, 3.16 U·mg{sup −1} enzyme). Furthermore, at an elevated temperature of 70 °C, immobilized PPL retained 60% of its initial activity. The PPL-MNPs nanocomposite was applied in the enzyme inhibition assays using orlistat, and two natural products isolated from oolong tea (i.e., EGCG and EGC) as the test compounds. - Highlights: • Porcine pancreatic lipase was firstly covalently immobilized onto carboxylated MNPs. • Immobilized porcine pancreatic lipase (PPL) was characterized by various techniques. • MNPs-PPL showed higher activity

  11. Effect of self-alkalization on nitrite accumulation in a high-rate denitrification system: Performance, microflora and enzymatic activities.

    Science.gov (United States)

    Li, Wei; Shan, Xiao-Yu; Wang, Zhi-Yao; Lin, Xiao-Yu; Li, Chen-Xu; Cai, Chao-Yang; Abbas, Ghulam; Zhang, Meng; Shen, Li-Dong; Hu, Zhi-Qiang; Zhao, He-Ping; Zheng, Ping

    2016-01-01

    The self-alkalization of denitrifying automatic circulation (DAC) reactor resulted in a large increase of pH up to 9.20 and caused a tremendous accumulation of nitrite up to 451.1 ± 49.0 mgN L(-1) at nitrate loading rate (NLR) from 35 kgN m(-3) d(-1) to 55 kgN m(-3) d(-1). The nitrite accumulation was greatly relieved even at the same NLR once the pH was maintained at 7.6 ± 0.2 in the system. Enzymatic assays indicated that the long-term bacterial exposure to high pH significantly inhibited the activity of copper type nitrite reductase (NirK) rather than the cytochrome cd1 type nitrite reductase (NirS). The terminal restriction fragment length polymorphism (T-RFLP) analysis revealed that the dominant denitrifying bacteria shifted from the NirS-containing Thauear sp. 27 to the NirK-containing Hyphomicrobium nitrativorans strain NL23 during the self-alkalization. The significant nitrite accumulation in the high-rate denitrification system could be therefore, due to the inhibition of Cu-containing NirK by high pH from the self-alkalization. The results suggest that the NirK-containing H. nitrativorans strain NL23 could be an ideal functional bacterium for the conversion of nitrate to nitrite, i.e. denitritation, which could be combined with anaerobic ammonium oxidation (Anammox) to develop a new process for nitrogen removal from wastewater. PMID:26595097

  12. Studies on gene structure, enzymatic activity and regulatory mechanism of acetohydroxy acid isomeroreductase from G2 pea

    Institute of Scientific and Technical Information of China (English)

    XU Yunjian (徐云剑); GU Xuesong (顾雪松); LI Jun (李珺); LI Qing (李 晴); Peter J. Davies; ZHU Yuxian (朱玉贤)

    2003-01-01

    The AAIR genomic DNA of G2 pea (Pisum sativum L.) was amplified by PCR method. Sequence analysis showed that it was composed of 8 introns and 9 exons with three of the introns containing specific A/T-rich endogenous promoter regions. Molecular hybridization experiments revealed that the expression of AAIR remained at a high level before and after flowering if grown in short day growth chambers. However, when grown under long day conditions, the level of AAIR expression declined very rapidly after flowering. This variation of AAIR expression is consistent with the change of enzymatic activity of acetohydroxy acid isomeroreductase. Functional complementation experiments carried out using an acetohydroxy acid isomeroreductase deficient E. coli strain showed that these cells could not grow on M9 medium without addition of branched-chain amino acids unless they were transformed with the AAIR expression vector. Further study revealed that overexpression of the pea AAIR cDNA in acetohydroxy acid isomeroreductase deficient E. coli strain enhanced significantly its branched-chain amino acid biosynthetic capacity. Results from gel shift experiments showed that fractions of pea nuclear protein extracts could bind specifically to some A/T rich regions present in introns of the AAIR gene. The A/T-rich-region-binding proteins remained at a steady level in the non-senescing apical buds of short-day grown G2 pea. In the rapid-senescing apical buds of long-day grown G2 pea, the levels of these proteins declined rapidly after flower initiation. Therefore, the nuclear protein binding capacities to endogenous promoter regions may constitute an important mechanism to regulate AAIR gene expression.

  13. Studies on the Effects of Polyaspartate Protease Fertilizer Enhancer in the Absorptions of Soil Nutrition and the Enzymatic Activities of Crops

    Institute of Scientific and Technical Information of China (English)

    JIANG Guoliang; YANG Dong; LIU Yun; ZHANG Guanghua; LI Zhongjun; ZHANG Xinhua

    2003-01-01

    The effects of polyaspartate protease fertilizer enhancer, made from oyster shell proteins, on the absorption of soil nutrition and the enzymatic activities of crops were studied. It has been found that the enhancer contributes 30%, 50 % and 50% augmentation of nitrogen (N), phosphate (P) and potassium (K) absorption respectively and about 20% of nitrate reductase and peroxide enzyme activities of crops. These results show that polyaspartate protease fertilizer enhancer could improve significantly the absorption and utilization efficiencies of soil nutrition and the activities of nitrate reductase and peroxide enzyme of crops, thus elevating the utilization rates of chemical fertilizers to a certain extent.

  14. Ultrasonic-assisted enzymatic extraction of phenolics from broccoli (Brassica oleracea L. var. italica) inflorescences and evaluation of antioxidant activity in vitro.

    Science.gov (United States)

    Wu, Hao; Zhu, Junxiang; Yang, Long; Wang, Ran; Wang, Chengrong

    2015-06-01

    An efficient ultrasonic-assisted enzymatic extraction technique was applied to extracting phenolics from broccoli inflorescences without organic solvents. The synergistic model of enzymolysis and ultrasonication simultaneously was selected, and the enzyme combination was optimized by orthogonal test: cellulase 7.5 mg/g FW (fresh weight), pectinase 10 mg/g FW, and papain 1.0 mg/g FW. The operating parameters in ultrasonic-assisted enzymatic extraction were optimized with response surface methodology using Box-Behnken design. The optimal extraction conditions were as follows: ultrasonic power, 440 W; liquid to material ratio, 7.0:1 mL/g; pH value of 6.0 at 54.5 ℃ for 10 min. Under these conditions, the extraction yield of phenolics achieved 1.816 ± 0.0187 mg gallic acid equivalents/gram FW. The free radical scavenging activity of ultrasonic-assisted enzymatic extraction extracts was determined by 1,1-diphenyl-2-picrylhydrazyl·assay with EC50 values of 0.25, and total antioxidant activity was determined by ferric reducing antioxidant power assay with ferric reducing antioxidant power value of 0.998 mmol FeSO4/g compared with the referential ascorbic acid of 1.184 mmol FeSO4/g. PMID:24837595

  15. Study on ACE-inhibitory and Antioxidant Activities of Mactra veneriformis Enzymatic Hydrolysis Peptides%四角蛤蜊酶解肽抑制ACE活性与抗氧化活性研究

    Institute of Scientific and Technical Information of China (English)

    朱蕴菡; 刘睿; 王令充; 贾梦蛟; 蒋金来; 刘新; 吴皓

    2013-01-01

    OBJECTIVE To evaluate the ACE-inhibitory and antioxidant activities of Mactra veneriformis enzymatic hydrolysis peptides. METHODS Pepsin and trypsin were used to simulate enzymatic process in vivo, and then enzymatic hydrolysis peptides of Mactra veneriformis were obtained. The ACE inhibitory activity of Mactra veneriformis enzymatic hydrolysis peptides was measured by high performance liquid chromatography, and the radical scavenging effect was detected by Fen ton system. RESULTS Mactra veneriformis enzymatic hydrolysis peptides was obtained by enzymatic hydrolysis of trypsin for 6h, which ACE-inhibitory activity of enzymatic hydrolysis peptides is the best, IC50 = 15. 06 μg/mL. The best rate of hydroxyl radical scavenging of Mactra veneriformis enzymatic hydrolysis peptides is 93. 78%, which was obtained by trypsin used for 2 h. CONCLUSION Mactra veneriformis enzymatic hydrolysis peptides have a definite ACE-inhibitory and antioxidant activity.%目的 评价四角蛤蜊酶解肽体外抑制ACE活性与抗氧化活性.方法 以胃蛋白酶和胰蛋白酶模拟在体过程对四角蛤蜊进行酶解获得酶解肽.利用高效液相色谱法测定四角蛤蜊酶解肽的抑制ACE的活性,以Fenton法检测四角蛤蜊酶解肽对羟自由基的清除作用.结果 胰蛋白酶酶解6h的四角蛤蜊酶解肽抑制ACE活性最佳,IC50=15.06μg/mL.胰蛋白酶酶解2h的羟自由基清除率最高,为93.78%.结论 四角蛤蜊酶解肽具有一定的抑制ACE活性与抗氧化活性.

  16. Contrasted enzymatic cocktails reveal the importance of cellulases and hemicellulases activity ratios for the hydrolysis of cellulose in presence of xylans.

    Science.gov (United States)

    Dondelinger, Eve; Aubry, Nathalie; Ben Chaabane, Fadhel; Cohen, Céline; Tayeb, Jean; Rémond, Caroline

    2016-03-01

    Various enzymatic cocktails were produced from two Trichoderma reesei strains, a cellulase hyperproducer strain and a strain with β-glucosidase activity overexpression. By using various carbon sources (lactose, glucose, xylose, hemicellulosic hydrolysate) for strains growth, contrasted enzymatic activities were obtained. The enzymatic cocktails presented various levels of efficiency for the hydrolysis of cellulose Avicel into glucose, in presence of xylans, or not. These latter were also hydrolyzed with different extents according to cocktails. The most efficient cocktails (TR1 and TR3) on Avicel were richer in filter paper activity (FPU) and presented a low ratio FPU/β-glucosidase activity. Cocktails TR2 and TR5 which were produced on the higher amount of hemicellulosic hydrolysate, possess both high xylanase and β-xylosidase activities, and were the most efficient for xylans hydrolysis. When hydrolysis of Avicel was conducted in presence of xylans, a decrease of glucose release occurred for all cocktails compared to hydrolysis of Avicel alone. Mixing TR1 and TR5 cocktails with two different ratios of proteins (1/1 and 1/4) resulted in a gain of efficiency for glucose release during hydrolysis of Avicel in presence of xylans compared to TR5 alone. Our results demonstrate the importance of combining hemicellulase and cellulase activities to improve the yields of glucose release from Avicel in presence of xylans. In this context, strategies involving enzymes production with carbon sources comprising mixed C5 and C6 sugars or combining different cocktails produced on C5 or on C6 sugars are of interest for processes developed in the context of lignocellulosic biorefinery. PMID:27001439

  17. Screening of mammary carcinoma for hormone dependency in vitro. Enzymatic activity in short-term organotypic cultures of breast biopsies from 62 patients.

    Science.gov (United States)

    Montessori, G A; Algard, F T; Van Netten, J P; Donald, J C

    1977-04-01

    Enzymatic activity in short-term organotypic cultures of breast biopsies from 62 patients. Am J Clin Pathol 67: 393-396, 1977. Mammary carcinomas from 62 patients were assessed for pentose shunt dehydrogenase activity initially and after 24-72 hours in organotypic cultures with or without exogenous hormones. Hormones tested were (1) estradiol, (2) testosterone, and (3) prolactin. Thirty-seven (60%) were judged hormone-independent, in vitro; 14 (23%) were judged hormone-dependent, in vitro; 11 (17%) were classed as "indeterminant." Clinical results of endocrine management of 13 cases and an appraisal of the usefulness of the method are presented. PMID:192068

  18. Effect of land-use types on soil enzymatic activities and chemical properties in semi-deciduous forest areas of Central-West Côte d'Ivoire

    Directory of Open Access Journals (Sweden)

    Gonnety, JT.

    2012-01-01

    Full Text Available Enzymatic activities play a key role in the biochemical functioning of soils. As a consequence, they have been proposed as indicators of soil quality. This study was conducted at the Oumé benchmark site (Central-West, Côte d'Ivoire, and aimed at measuring the enzymatic activities involved in the phosphorus (acid phosphatase and alkaline phosphatase, nitrogen (N-acetyl-β-D glucosaminidase and carbon (β-glucosidase and N-acetyl-β-D glucosaminidase cycles. Soil from four main agro-ecological units (a secondary forest, a 20 year-old cocoa plantation, a 2 year-old Chromolaena odorata-based fallow and a continuous maize crop, representative of land-use systems in the area, were sampled for the measurement of enzymatic activities and chemical characteristics. Results showed that the enzymatic activity values were the highest in the fallow soil, whereas the maize crop displayed the lowest levels of enzymatic activity in soil. Moreover, soil from C. odorata fallow displayed the highest values of C, N, exchangeable bases (Mg2+, K+ contents, and CEC, and the lowest C:N ratio, which are characteristics of good quality soil. A Principal Component Analysis revealed a marked relationship between C, N and enzymatic activity levels, showing that these enzymes are suitable for monitoring soil quality in semi-deciduous forest areas in Central-West Côte d'Ivoire.

  19. pCO2 and enzymatic activity in a river floodplain system of the Danube under different hydrological settings.

    Science.gov (United States)

    Sieczko, Anna; Demeter, Katalin; Mayr, Magdalena; Meisterl, Karin; Peduzzi, Peter

    2014-05-01

    Surface waters may serve as either sinks or sources of CO2. In contrast to rivers, which are typically sources of CO2 to the atmosphere, the role of fringing floodplains in CO2 flux is largely understudied. This study was conducted in a river-floodplain system near Vienna (Austria). The sampling focused on changing hydrological situations, particularly on two distinct flood events: a typical 1-year flood in 2012 and an extraordinary 100-year flood in 2013. One objective was to determine partial pressure of CO2 (pCO2) in floodplain lakes with different degree of connectivity to the main channel, and compare the impact of these two types of floods. Another aim was to decipher which fraction of the dissolved organic matter (DOM) pool contributed to pCO2 by linking pCO2 with optical properties of DOM and extracellular enzymatic activity (EEA) of microbes. The EEA is a valuable tool, especially for assessing the non-chromophoric but rapidly utilized DOM-fraction during floods. In 2012 and 2013, the floodplain lakes were dominated by supersaturated pCO2 conditions, which indicates that they served as CO2 sources. Surprisingly, there were no significant differences in pCO2 between the two types of flood. Our findings imply that the extent of the flood had minor impact on pCO2, but the general occurrence of a flood appears to be important. During the flood in 2013 significantly more dissolved organic carbon (DOC) (penzymes (EEA-C) was observed. We also found positive correlations of pCO2with EEA-C both in 2012 (r=0.86, p<0.01) and in 2013 (r=0.73, p<0.05). The above findings imply that some fraction of DOM, which was introduced during the floods, was reactive and could be utilized by prokaryotes. This also indicates that pCO2 during floods was driven by degradation of e.g. carbohydrates.

  20. Modulation of the pharmacological effects of enzymatically-active PLA2 by BTL-2, an isolectin isolated from the Bryothamnion triquetrum red alga

    Directory of Open Access Journals (Sweden)

    Nagano Celso S

    2008-06-01

    Full Text Available Abstract Background An interaction between lectins from marine algae and PLA2 from rattlesnake was suggested some years ago. We, herein, studied the effects elicited by a small isolectin (BTL-2, isolated from Bryothamnion triquetrum, on the pharmacological and biological activities of a PLA2 isolated from rattlesnake venom (Crotalus durissus cascavella, to better understand the enzymatic and pharmacological mechanisms of the PLA2 and its complex. Results This PLA2 consisted of 122 amino acids (approximate molecular mass of 14 kDa, its pI was estimated to be 8.3, and its amino acid sequence shared a high degree of similarity with that of other neurotoxic and enzymatically-active PLA2s. BTL-2 had a molecular mass estimated in approximately 9 kDa and was characterized as a basic protein. In addition, BTL-2 did not exhibit any enzymatic activity. The PLA2 and BTL-2 formed a stable heterodimer with a molecular mass of approximately 24–26 kDa, estimated by molecular exclusion HPLC. In the presence of BTL-2, we observed a significant increase in PLA2 activity, 23% higher than that of PLA2 alone. BTL-2 demonstrated an inhibition of 98% in the growth of the Gram-positive bacterial strain, Clavibacter michiganensis michiganensis (Cmm, but only 9.8% inhibition of the Gram-negative bacterial strain, Xanthomonas axonopodis pv passiflorae (Xap. PLA2 decreased bacterial growth by 27.3% and 98.5% for Xap and Cmm, respectively, while incubating these two proteins with PLA2-BTL-2 inhibited their growths by 36.2% for Xap and 98.5% for Cmm. PLA2 significantly induced platelet aggregation in washed platelets, whereas BTL-2 did not induce significant platelet aggregation in any assay. However, BTL-2 significantly inhibited platelet aggregation induced by PLA2. In addition, PLA2 exhibited strong oedematogenic activity, which was decreased in the presence of BTL-2. BTL-2 alone did not induce oedema and did not decrease or abolish the oedema induced by the 48

  1. Highly Sensitive Spectrofluorimetric Determination of Riboflavin Based on the Generation of Active Oxygen Coupled with Enzymatic Reaction

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A spectrofluorimetric method for the determining riboflavin (VB2) based on its enhancement on the fluorescence of hemoglobin-catalyzed enzymatic reaction was proposed. The proposed method consisted of two reactions. One was the photochemical reaction of VB2, the other was a hemoglobin-catalyzed enzymatic reaction. The optimal experimental conditions for the determinations were established. The linear range of the method was 5.0×10-9-1.0×10-7mol/L of VB2. The detection limit was calculated to be 3.65×10-9 mol/L. The relative standard deviation of this method was 2.3 % at 7.0×10-8 mol/L for 11 determinations.

  2. 鳙鱼活性多肽酶法制备工艺研究%Enzymatic Extraction of Active Polypeptide from Hypophthalmichthys nobilis

    Institute of Scientific and Technical Information of China (English)

    张雪; 陈复生; 隋继学

    2013-01-01

      为优化鳙鱼活性多肽酶法制备工艺,分析了鳙鱼肉糜预处理温度和酶解温度对水解度的影响,确定了最佳的预处理条件为85℃水浴中加热预处理20 min,酶解温度设为55~75℃.经均匀设计实验优选和最优条件验证实验证实,以氮溶指数为指标的最优酶解条件为:酶解时间8.0 h,固液比1∶4.25,蛋白酶A用量3‰,酶解温度75℃,产物氮溶指数达80.54%;以多肽得率为指标的最优酶解条件为:酶解时间8.0 h,固液比1∶2,蛋白酶A用量3‰,酶解温度75℃,产物多肽得率达11.92%;以产物总抗氧化指数为指标的最优酶解条件为:酶解时间1.0 h,固液比1∶6,蛋白酶A用量3‰,酶解温度55℃,所得产物总抗氧化指数达87.42‰.%For optimizating the enzymatic extraction of active polypeptide from Hypophthalmichthys nobilis,we investigated the effects of minced meat pretreatment temperature and enzymatic temperature on the degree of hydrolysis,and determined that the optimal pretreatment conditions was in a water bath of 85℃to heat pretreatment 20 min. Through uniform design and verification experiment,we confirmed that the optimal enzymatic hydrolysis conditions by using optimal conditionsuse nitrogen solubility index as an indicator were enzymatic hydrolysis time 8.0 h,the solid-liquid ratio 1∶4.25,dosage of protease A 3‰,hydrolysis temperature 75℃,then nitrogen solubility index of the product reached 80.54%. The optimal hydrolysis conditions of using polypeptide yield as indicators were enzymatic hydrolysis time 8.0 h,solid-liquid ratio of 1∶2,dosage of protease A 3‰,the temperature of enzymatic hydrolysis 75℃,then the polypeptide yield of product reached 11.92%. The optimal enzymatic hydrolysis conditions of using the total antioxidant index of the product as indicators were the enzymatic time 1.0 h,solid-liquid ratio 1∶6,dosage of protease A 3‰,hydrolysis temperature 55 ℃,and then the total antioxidant index

  3. Enzymatic Hydrolysis of Salmon By-products: Effect of Process Conditions on ACE Inhibiting Activities of Fish Protein Hydrolysates

    OpenAIRE

    Five, Kathrine

    2013-01-01

    By-products from the salmon farming industry contain valuable components, such as proteins and lipids. By-products like frames, heads and viscera can be used as raw material for the production of fish protein hydrolysates with high nutritional value, but also bioactive properties. The hydrolysates are produced by enzymatic hydrolysis using endogenous and commercial enzymes, and the process conditions and raw material influence the properties of the hydrolysate. The first aim of this thesis wa...

  4. Impact of the redox-cycling herbicide diquat on transcript expression and antioxidant enzymatic activities of the freshwater snail Lymnaea stagnalis

    International Nuclear Information System (INIS)

    The presence of pesticides in the environment results in potential unwanted effects on non-target species. Freshwater organisms inhabiting water bodies adjacent to agricultural areas, such as ditches, ponds and marshes, are good models to test such effects as various pesticides may reach these habitats through several ways, including aerial drift, run-off, and drainage. Diquat is a non-selective herbicide used for crop protection or for weed control in such water bodies. In this study, we investigated the effects of diquat on a widely spread aquatic invertebrate, the holarctic freshwater snail Lymnaea stagnalis. Due to the known redox-cycling properties of diquat, we studied transcript expression and enzymatic activities relative to oxidative and general stress in the haemolymph and gonado-digestive complex (GDC). As diquat is not persistent, snails were exposed for short times (5, 24, and 48 h) to ecologically relevant concentrations (22.2, 44.4, and 222.2 μg l−1) of diquat dibromide. RT-qPCR was used to quantify the transcription of genes encoding catalase (cat), a cytosolic superoxide dismutase (Cu/Zn-sod), a selenium-dependent glutathione peroxidase (gpx), a glutathione reductase (gred), the retinoid X receptor (rxr), two heat shock proteins (hsp40 and hsp70), cortactin (cor) and the two ribosomal genes r18S and r28s. Enzymatic activities of SOD, Gpx, Gred and glutathione S-transferase (GST) were investigated in the GDC using spectrophoto/fluorometric methods. Opposite trends were obtained in the haemolymph depending on the herbicide concentration. At the lowest concentration, effects were mainly observed after 24 h of exposure, with over-transcription of cor, hsp40, rxr, and sod, whereas higher concentrations down-regulated the expression of most of the studied transcripts, especially after 48 h of exposure. In the GDC, earlier responses were observed and the fold-change magnitude was generally much higher: transcription of all target genes increased

  5. Impact of the redox-cycling herbicide diquat on transcript expression and antioxidant enzymatic activities of the freshwater snail Lymnaea stagnalis

    Energy Technology Data Exchange (ETDEWEB)

    Bouetard, Anthony, E-mail: anthony.bouetard@rennes.inra.fr [INRA, UMR INRA-Agrocampus Ouest ESE 0985, Equipe Ecotoxicologie et Qualite des Milieux Aquatiques, 65 rue de Saint-Brieuc, 35042 Rennes cedex (France); Besnard, Anne-Laure; Vassaux, Daniele; Lagadic, Laurent; Coutellec, Marie-Agnes [INRA, UMR INRA-Agrocampus Ouest ESE 0985, Equipe Ecotoxicologie et Qualite des Milieux Aquatiques, 65 rue de Saint-Brieuc, 35042 Rennes cedex (France)

    2013-01-15

    The presence of pesticides in the environment results in potential unwanted effects on non-target species. Freshwater organisms inhabiting water bodies adjacent to agricultural areas, such as ditches, ponds and marshes, are good models to test such effects as various pesticides may reach these habitats through several ways, including aerial drift, run-off, and drainage. Diquat is a non-selective herbicide used for crop protection or for weed control in such water bodies. In this study, we investigated the effects of diquat on a widely spread aquatic invertebrate, the holarctic freshwater snail Lymnaea stagnalis. Due to the known redox-cycling properties of diquat, we studied transcript expression and enzymatic activities relative to oxidative and general stress in the haemolymph and gonado-digestive complex (GDC). As diquat is not persistent, snails were exposed for short times (5, 24, and 48 h) to ecologically relevant concentrations (22.2, 44.4, and 222.2 {mu}g l{sup -1}) of diquat dibromide. RT-qPCR was used to quantify the transcription of genes encoding catalase (cat), a cytosolic superoxide dismutase (Cu/Zn-sod), a selenium-dependent glutathione peroxidase (gpx), a glutathione reductase (gred), the retinoid X receptor (rxr), two heat shock proteins (hsp40 and hsp70), cortactin (cor) and the two ribosomal genes r18S and r28s. Enzymatic activities of SOD, Gpx, Gred and glutathione S-transferase (GST) were investigated in the GDC using spectrophoto/fluorometric methods. Opposite trends were obtained in the haemolymph depending on the herbicide concentration. At the lowest concentration, effects were mainly observed after 24 h of exposure, with over-transcription of cor, hsp40, rxr, and sod, whereas higher concentrations down-regulated the expression of most of the studied transcripts, especially after 48 h of exposure. In the GDC, earlier responses were observed and the fold-change magnitude was generally much higher: transcription of all target genes increased

  6. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    -efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through the enzymatic hydrolysis of sphingomyelin have been studied. sphingomyelin is a ubiquitous membrane-lipid and rich in dairy products or by-products. It has been verified that...... sphingomyelin modification gives a feasible approach to the potential production of ceramide. The reaction system has been improved through system evaluation and the optimization of several important factors, and phospholipase C from Clostridium perfringens shows higher activity towards the hydrolysis reaction...

  7. The Membrane-anchored Serine Protease Prostasin (CAP1/PRSS8) Supports Epidermal Development and Postnatal Homeostasis Independent of Its Enzymatic Activity

    DEFF Research Database (Denmark)

    Peters, Diane E; Szabo, Roman; Friis, Stine;

    2014-01-01

    prostasin catalytically inactive (Prss8(Cat-/Cat-) mice), develop barrier function and are healthy when followed for up to 20 weeks. This striking difference could not be explained by genetic modifiers or by maternal effects, as these divergent phenotypes were displayed by Prss8(-/-) and Prss8(Cat-/Cat......-) mice born within the same litter. Furthermore, Prss8(Cat-/Cat-) mice were able to regenerate epidermal covering following cutaneous wounding. This study provides the first demonstration that essential in vivo functions of prostasin are executed by a non-enzymatic activity of this unique membrane...

  8. Defectively N-glycosylated and non-O-glycosylated aminopeptidase N (CD13) is normally expressed at the cell surface and has full enzymatic activity

    DEFF Research Database (Denmark)

    Norén, K; Hansen, Gert Helge; Clausen, H;

    1997-01-01

    the glycoprotein aminopeptidase N can be synthesized and the effects of altered glycosylation can be studied. It is demonstrated that aminopeptidase N carries "mucin-type" O-glycans and that this is predominantly located in the stalk, which connects the catalytic headgroup to the membrane anchor...... changes in N-glycosylation as the various glycosylated forms of aminopeptidase N are normally converted from the high-mannose form to the complex glycosylated form. Enzymatic activity is not influenced by the changes in glycosylation....

  9. Monitoring enzymatic ATP hydrolysis by EPR spectroscopy

    OpenAIRE

    Hacker, Stephan M.; Hintze, Christian; Marx, Andreas; Drescher, Malte

    2014-01-01

    An adenosine triphosphate (ATP) analogue modified with two nitroxide radicals is developed and employed to study its enzymatic hydrolysis by electron paramagnetic resonance spectroscopy. For this application, we demonstrate that EPR holds the potential to complement fluorogenic substrate analogues in monitoring enzymatic activity.

  10. The stay-green phenotype of TaNAM-RNAi wheat plants is associated with maintenance of chloroplast structure and high enzymatic antioxidant activity.

    Science.gov (United States)

    Checovich, Mariana L; Galatro, Andrea; Moriconi, Jorge I; Simontacchi, Marcela; Dubcovsky, Jorge; Santa-María, Guillermo E

    2016-07-01

    TaNAM transcription factors play an important role in controlling senescence, which in turn, influences the delivery of nitrogen, iron and other elements to the grain of wheat (Triticum aestivum) plants, thus contributing to grain nutritional value. While lack or diminished expression of TaNAMs determines a stay-green phenotype, the precise effect of these factors on chloroplast structure has not been studied. In this work we focused on the events undergone by chloroplasts in two wheat lines having either control or diminished TaNAM expression due to RNA interference (RNAi). It was found that in RNAi plants maintenance of chlorophyll levels and maximal photochemical efficiency of photosystem II were associated with lack of chloroplast dismantling. Flow cytometer studies and electron microscope analysis showed that RNAi plants conserved organelle ultrastructure and complexity. It was also found that senescence in control plants was accompanied by a low leaf enzymatic antioxidant activity. Lack of chloroplast dismantling in RNAi plants was associated with maintenance of protein and iron concentration in the flag leaf, the opposite being observed in control plants. These data provide a structural basis for the observation that down regulation of TaNAMs confers a functional stay-green phenotype and indicate that the low export of iron and nitrogen from the flag leaf of these plants is concomitant, within the developmental window studied, with lack of chloroplast degradation and high enzymatic antioxidant activity. PMID:27061370

  11. Mutations in Mtr4 Structural Domains Reveal Their Important Role in Regulating tRNAiMet Turnover in Saccharomyces cerevisiae and Mtr4p Enzymatic Activities In Vitro.

    Directory of Open Access Journals (Sweden)

    Yan Li

    Full Text Available RNA processing and turnover play important roles in the maturation, metabolism and quality control of a large variety of RNAs thereby contributing to gene expression and cellular health. The TRAMP complex, composed of Air2p, Trf4p and Mtr4p, stimulates nuclear exosome-dependent RNA processing and degradation in Saccharomyces cerevisiae. The Mtr4 protein structure is composed of a helicase core and a novel so-called arch domain, which protrudes from the core. The helicase core contains highly conserved helicase domains RecA-1 and 2, and two structural domains of unclear functions, winged helix domain (WH and ratchet domain. How the structural domains (arch, WH and ratchet domain coordinate with the helicase domains and what roles they are playing in regulating Mtr4p helicase activity are unknown. We created a library of Mtr4p structural domain mutants for the first time and screened for those defective in the turnover of TRAMP and exosome substrate, hypomodified tRNAiMet. We found these domains regulate Mtr4p enzymatic activities differently through characterizing the arch domain mutants K700N and P731S, WH mutant K904N, and ratchet domain mutant R1030G. Arch domain mutants greatly reduced Mtr4p RNA binding, which surprisingly did not lead to significant defects on either in vivo tRNAiMet turnover, or in vitro unwinding activities. WH mutant K904N and Ratchet domain mutant R1030G showed decreased tRNAiMet turnover in vivo, as well as reduced RNA binding, ATPase and unwinding activities of Mtr4p in vitro. Particularly, K904 was found to be very important for steady protein levels in vivo. Overall, we conclude that arch domain plays a role in RNA binding but is largely dispensable for Mtr4p enzymatic activities, however the structural domains in the helicase core significantly contribute to Mtr4p ATPase and unwinding activities.

  12. Enzymatic activity in turkey, duck, quail and chicken liver microsomes against four human cytochrome P450 prototype substrates and aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Hansen W. Murcia

    2011-10-01

    Full Text Available Cytochrome P450 enzymes (CYP are a group of monooxygenases able to biotransform several kinds of xenobiotics including aflatoxin B1 (AFB1, a highly toxic mycotoxin. These enzymes have been widely studied in humans and others mammals, but there is not enough information in commercial poultry species about their biochemical characteristics or substrate specificity. The aim of the present study was to identify CYPs from avian liver microsomes with the use of prototype substrates specific for human CYP enzymes and AFB1. Biochemical characterization was carried out in vitro and biotransformation products were detected by high-performance liquid chromatography (HPLC. Enzymatic constants were calculated and comparisons between turkey, duck, quail and chicken activities were done. The results demonstrate the presence of four avian ortholog enzyme activities possibly related with a CYP1A1, CYP1A2, CYP2A6 (activity not previously identified and CYP3A4 poultry orthologs, respectively. Large differences in enzyme kinetics specific for prototype substrates were found among the poultry species studied. Turkey liver microsomes had the highest affinity and catalytic rate for AFB1 whereas chicken enzymes had the lowest affinity and catalytic rate for the same substrate. Quail and duck microsomes showed intermediate values. These results correlate well with the known in vivo sensitivity for AFB1 except for the duck. A high correlation coefficient between 7-ethoxyresorufin-Odeethylase (EROD and 7-methoxyresorufin- O-deethylase (MROD activities was found in the four poultry species, suggesting that these two enzymatic activities might be carried out by the same enzyme. The results of the present study indicate that four prototype enzyme activities are present in poultry liver microsomes, possibly related with the presence of three CYP avian orthologs. More studies are needed in order to further characterize these enzymes.

  13. Enzymatic Treatment of Whey Proteins in Cow's Milk Results in Differential Inhibition of IgE-Mediated Mast Cell Activation Compared to T-Cell Activation

    NARCIS (Netherlands)

    Knipping, Karen; van Esch, Betty C. A. M.; van Ieperen-van Dijk, Adrie G.; van Hoffen, Els; van Baalen, Ton; Knippels, Leon M. J.; van der Heide, Sicco; Dubois, Anthony E. J.; Garssen, Johan; Knol, Edward F.

    2012-01-01

    Background: Cow's milk (CM) hydrolysates are frequently used as milk substitutes for children with CM allergy. In hydrolysates, allergenic epitopes within CM proteins are diminished by enzymatic treatment. The aim of this study was to examine the allergenic and immunogenic properties of whey protein

  14. A δ38 deletion variant of human transketolase as a model of transketolase-like protein 1 exhibits no enzymatic activity.

    Directory of Open Access Journals (Sweden)

    Stefan Schneider

    Full Text Available Besides transketolase (TKT, a thiamin-dependent enzyme of the pentose phosphate pathway, the human genome encodes for two closely related transketolase-like proteins, which share a high sequence identity with TKT. Transketolase-like protein 1 (TKTL1 has been implicated in cancerogenesis as its cellular expression levels were reported to directly correlate with invasion efficiency of cancer cells and patient mortality. It has been proposed that TKTL1 exerts its function by catalyzing an unusual enzymatic reaction, a hypothesis that has been the subject of recent controversy. The most striking difference between TKTL1 and TKT is a deletion of 38 consecutive amino acids in the N-terminal domain of the former, which constitute part of the active site in authentic TKT. Our structural and sequence analysis suggested that TKTL1 might not possess transketolase activity. In order to test this hypothesis in the absence of a recombinant expression system for TKTL1 and resilient data on its biochemical properties, we have engineered and biochemically characterized a "pseudo-TKTL1" Δ38 deletion variant of human TKT (TKTΔ38 as a viable model of TKTL1. Although the isolated protein is properly folded under in vitro conditions, both thermal stability as well as stability of the TKT-specific homodimeric assembly are markedly reduced. Circular dichroism and NMR spectroscopic analysis further indicates that TKTΔ38 is unable to bind the thiamin cofactor in a specific manner, even at superphysiological concentrations. No transketolase activity of TKTΔ38 can be detected for conversion of physiological sugar substrates thus arguing against an intrinsically encoded enzymatic function of TKTL1 in tumor cell metabolism.

  15. Enzymatic modification of starch

    DEFF Research Database (Denmark)

    Jensen, Susanne Langgård

    In the food industry approaches for using bioengineering are investigated as alternatives to conventional chemical and physical starch modification techniques in development of starches with specific properties. Enzyme-assisted post-harvest modification is an interesting approach to this, since it...... is considered a clean and energy saving technology. This thesis aimed to investigate the effect of using reaction conditions, simulating an industrial process, for enzymatic treatment of starch with branching enzyme (BE) from Rhodothermus obamensis. Thus treatements were conducted at 70°C using very...... high substrate concentration (30-40% dry matter (DM)) and high enzyme activity (750-2250 BE units (BEU)/g sample). Starches from various botanical sources, representing a broad range of properties, were used as substrates. The effects of the used conditions on the BE-reaction were evaluated by...

  16. Effect of land-use types on soil enzymatic activities and chemical properties in semi-deciduous forest areas of Central-West Côte d'Ivoire

    OpenAIRE

    Gonnety, JT.; Assémien, EFL.; Guéi, AM.; N'Dri, AA.; Djina, Y.; Koné, AW.; Tondoh, JE.

    2012-01-01

    Enzymatic activities play a key role in the biochemical functioning of soils. As a consequence, they have been proposed as indicators of soil quality. This study was conducted at the Oumé benchmark site (Central-West, Côte d'Ivoire), and aimed at measuring the enzymatic activities involved in the phosphorus (acid phosphatase and alkaline phosphatase), nitrogen (N-acetyl-β-D glucosaminidase) and carbon (β-glucosidase and N-acetyl-β-D glucosaminidase) cycles. Soil from four main agro-ecological...

  17. Enzymatic sulfation of gastric mucous glycoprotein in rat--changes in glycoprotein sulfotransferase activity with stress and anti-ulcer agent, sofalcone

    International Nuclear Information System (INIS)

    Enzymatic sulfation of mucous glycoprotein (GP) was studied in gastric mucosa of rat. After rat stomach was incubated with [35S]-sulfate, incorporation of radioactivity into gastric mucosal APS (adenosine 5'-phosphosulfate), PAPS (3'-phosphoadenosine 5'-phosphosulfate) and endogenous GPs could be detected. The degree of sulfation of endogenous GPs was highest in the macromolecular GP (peak I) and lowest in the low molecular GP (peak III). By using a crude preparation of GP sulfotransferase from rat gastric mucosa, the transfer of [35S]-sulfate from [35S]-PAPS into macromolecular mucous GP was determined as being the activity of sulfotransferase. The activity of GP sulfotransferase was mainly distributed in the microsomal fraction, and was proportional to the incubation time, substrate (mucous GP) concentration and [35S]-PAPS concentration. The enzyme activity was significantly higher in the corpus than that in the antral mucosa. The activity of GP sulfotransferase was significantly decreased at 6 h and was significantly increased at 12 h after the stress load, compared with that of the non-stressed condition. Anti-ulcer agent, sofalcone, increased the GP sulfotransferase activity under the stressed condition. On the other hand, cimetidine showed a significant inhibitory effect under the same condition. Changes in the GP sulfotransferase activity with stress and anti-ulcer agents were consistent with those in the incorporation of [35S]-sulfate into macromolecular mucous GP. These results suggest the importance of GP sulfotransferase as a key enzyme regulating the sulfation of mucous GP

  18. Changes of apparent enzymatic activities and physical and chemical properties of steam-exploded corn stover during enzymatic hydrolysis%汽爆玉米秸秆酶解时表观酶活及物料理化性质的变化

    Institute of Scientific and Technical Information of China (English)

    张霞; 李红伟; 马晓建

    2013-01-01

    Based on engineering practice, the sugar concentration, power and apparent enzymatic activities change of enzymolysis suspension in enzymolysis process of steam-exploded corn stover (SECS) were studied to promote the industrialization process of corn stover producing butanol. At the same time, the changes of SECS physical and chemical properties during enzymatic hydrolysis were studied by biological microscope, laser scattering particle size distribution analysis (LSPSDA), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Enzymatic hydrolysis was performed in a reactor with standard commercial cellulase as the enzyme. Enzymatic hydrolysis took place at 50℃, 100 r/min (four oblique leaves T agitator), enzyme loading of 60 IU/g corn stover, a solid:liquid ratio 3:10, and 48 h duration. Before the enzyme was added, the pH of the residues was adjusted to 4.8 with sodium hydroxide. After enzymatic hydrolysis the samples were taken from the reactor and centrifuged, and the supernatant phase was collected and analyzed for sugar concentrations, cellulase apparent enzymatic activities, and the physical and chemical properties of SECS. Sugar concentrations were tested by 3,5-Dinitrosalicylicacid(DNS). Cellulase apparent enzymatic activities were determined by the filter-paper method. Scanning electron microscopy and X-ray diffraction analysis were conducted. The results show: In the first reaction, the sugar concentration in the enzymatic hydrolysis liquid increased very quickly, especially in the first hour. The change in sugar concentration was not very clear later in the hydrolysis reaction due to such factors as substrate loss, changes in substrate properties, cellulose inactivation, decreasing cellulose synergy, and so on. The sugar concentration after enzymatic hydrolysis 48 h was 56.25 g/L. The stirring power decreased during enzymatic hydrolysis and declined most quickly after one hour of enzymatic hydrolysis, then began to decline more slowly. The

  19. Urbanised beaches of the Ligurian coastal area (NW Mediterranean): a classification based on organic-matter characteristics and hydrolytic enzymatic activities.

    Science.gov (United States)

    Misic, Cristina; Covazzi Harriague, Anabella

    2013-01-01

    The beaches of Liguria have been intensively affected by human activities for over a century, transforming nearly the entire coastline from natural to urbanised and significantly upsetting beach ecological properties. The present study aims to investigate 9 Ligurian beaches characterised by different degree of urbanisation, to test if and to what extent the organic-matter (OM) recycling processes can be linked to the human activity. Swash zone sediment, sampled during the spring-summer-autumn period, when the anthropogenic influence is at its maximum due to tourism, was analysed for OM features and recycling processes. Multivariate statistical analyses showed that huge amounts of detrital OM accumulated in the more urbanised sites, where the anthropogenic influence was at its peak, deriving from higher inhabitant number and density, from the presence of crowded roads very near to the swash zone and sewage treatment plants. The presence of torrent outlets on the beaches provided further OM accumulation. Lipids, carbohydrates and degraded autotrophic pigments were the OM fractions mainly responsible of the differentiation, and rather constant, high labile phosphorus contents were found in the more urbanised sites. The high activity values of the hydrolytic enzymes indicate the response of the microbial system to the OM accumulation in the urban sites. However, a decoupling of the trends of some enzymatic activities (namely glucosidase and lipase) and their target OM was observed in the highly urbanised conditions. PMID:23660184

  20. Enzymatic conversion of carbon dioxide.

    Science.gov (United States)

    Shi, Jiafu; Jiang, Yanjun; Jiang, Zhongyi; Wang, Xueyan; Wang, Xiaoli; Zhang, Shaohua; Han, Pingping; Yang, Chen

    2015-10-01

    With the continuous increase in fossil fuels consumption and the rapid growth of atmospheric CO2 concentration, the harmonious state between human and nature faces severe challenges. Exploring green and sustainable energy resources and devising efficient methods for CO2 capture, sequestration and utilization are urgently required. Converting CO2 into fuels/chemicals/materials as an indispensable element for CO2 capture, sequestration and utilization may offer a win-win strategy to both decrease the CO2 concentration and achieve the efficient exploitation of carbon resources. Among the current major methods (including chemical, photochemical, electrochemical and enzymatic methods), the enzymatic method, which is inspired by the CO2 metabolic process in cells, offers a green and potent alternative for efficient CO2 conversion due to its superior stereo-specificity and region/chemo-selectivity. Thus, in this tutorial review, we firstly provide a brief background about enzymatic conversion for CO2 capture, sequestration and utilization. Next, we depict six major routes of the CO2 metabolic process in cells, which are taken as the inspiration source for the construction of enzymatic systems in vitro. Next, we focus on the state-of-the-art routes for the catalytic conversion of CO2 by a single enzyme system and by a multienzyme system. Some emerging approaches and materials utilized for constructing single-enzyme/multienzyme systems to enhance the catalytic activity/stability will be highlighted. Finally, a summary about the current advances and the future perspectives of the enzymatic conversion of CO2 will be presented. PMID:26055659

  1. Chemo-enzymatic synthesis of a series of 2,4-syn-functionalized (S)-glutamate analogues: new insight into the structure-activity relation of ionotropic glutamate receptor subtypes 5, 6, and 7

    DEFF Research Database (Denmark)

    Sagot, Emanuelle; Pickering, Darryl S; Pu, Xiaosui;

    2008-01-01

    ( S)-Glutamic acid (Glu) is the major excitatory neurotransmitter in the central nervous system (CNS) activating the plethora of ionotropic Glu receptors (iGluRs) and metabotropic Glu receptors (mGluRs). In this paper, we present a chemo-enzymatic strategy for the enantioselective synthesis of fi...

  2. Propeptide of carboxypeptidase Y provides a chaperone-like function as well as inhibition of the enzymatic activity

    DEFF Research Database (Denmark)

    Winther, Jakob R.; Sørensen, P

    1991-01-01

    The zymogen of the vacuolar carboxypeptidase Y from Saccharomyces cerevisiae was purified and characterized with respect to activation as well as refolding in vitro. The purified procarboxypeptidase Y has no detectable activity but can be efficiently activated by proteinase K from Tritirachium al...

  3. Enzymatic reactions in dense gases

    OpenAIRE

    Knez, Željko

    2012-01-01

    The developments on applications of supercritical fluids as alternative solvents for biocatalytic processes that have taken place over the past two decades have been reviewed. An overview of process parameters influencing enzyme activity and stability, the influence of process parameters on reaction rates and productivity are presented. Applications of various types of reactors for enzymatic reaction in dense fluids, limitations of using enzymes as biocatalyst in supercritical fluids as well ...

  4. Analysis of Non-Enzymatically Glycated Peptides: Neutral-Loss Triggered MS3 Versus Multi-Stage Activation Tandem Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qibin; Petyuk, Vladislav A.; Schepmoes, Athena A.; Orton, Daniel J.; Monroe, Matthew E.; Yang, Feng; Smith, Richard D.; Metz, Thomas O.

    2008-10-15

    Non-enzymatic glycation of tissue proteins has important implications in the development of complications of diabetes mellitus. While electron transfer dissociation (ETD) has been shown to outperform collision-induced dissociation (CID) in sequencing glycated peptides by tandem mass spectrometry, ETD instrumentation is not yet available in all laboratories. In this study, we evaluated different advanced CID techniques (i.e., neutral-loss triggered MS3 and multi-stage activation) during LC-MSn analyses of Amadori-modified peptides enriched from human serum glycated in vitro. During neutral-loss triggered MS3 experiments, MS3 scans triggered by neutral-losses of 3 H2O or 3 H2O + HCHO produced similar results in terms of glycated peptide identifications. However, neutral losses of 3 H2O resulted in significantly more glycated peptide identifications during multi-stage activation experiments. Overall, the multi-stage activation approach produced more glycated peptide identifications, while the neutral-loss triggered MS3 approach resulted in much higher specificity. Both techniques offer a viable alternative to ETD for identifying glycated peptides when that method is unavailable.

  5. The role of the C-terminal region on the oligomeric state and enzymatic activity of Trypanosoma cruzi hypoxanthine phosphoribosyl transferase.

    Science.gov (United States)

    Valsecchi, Wanda M; Cousido-Siah, Alexandra; Defelipe, Lucas A; Mitschler, André; Podjarny, Alberto; Santos, Javier; Delfino, José M

    2016-06-01

    Hypoxanthine phosphoribosyl transferase from Trypanosoma cruzi (TcHPRT) is a critical enzyme for the survival of the parasite. This work demonstrates that the full-length form in solution adopts a stable and enzymatically active tetrameric form, exhibiting large inter-subunit surfaces. Although this protein irreversibly aggregates during unfolding, oligomerization is reversible and can be modulated by low concentrations of urea. When the C-terminal region, which is predicted as a disordered stretch, is excised by proteolysis, TcHPRT adopts a dimeric state, suggesting that the C-terminal region acts as a main guide for the quaternary arrangement. These results are in agreement with X-ray crystallographic data presented in this work. On the other hand, the C-terminal region exhibits a modulatory role on the enzyme, as attested by the enhanced activity observed for the dimeric form. Bisphosphonates act as substrate-mimetics, uncovering long-range communications among the active sites. All in all, this work contributes to establish new ways applicable to the design of novel inhibitors that could eventually result in new drugs against parasitic diseases. PMID:26969784

  6. Chemo-Enzymatic Synthesis of Optically Active γ- and δ-Decalactones and Their Effect on Aphid Probing, Feeding and Settling Behavior.

    Directory of Open Access Journals (Sweden)

    Filip Boratyński

    Full Text Available The enantiomerically enriched γ- and δ-decalactones (4a and 4b were prepared from corresponding racemic primary-secondary 1,4- and 1,5-diols (1a and 1b, as products of enzymatic oxidation catalyzed by different alcohol dehydrogenases. The results of biotransformations indicated that the oxidation processes catalyzed by alcohol dehydrogenase (HLADH, both isolated from horse liver and recombinant in Escherichia coli, were characterized by the highest degree of conversion with moderate enantioselectivity of the reaction. Useful, environmentally friendly extraction procedure of decalactones (4a and 4b based on hydrodistillation using a Deryng apparatus was developed. Both racemic lactones (4a and 4b, as well as their enantiomerically enriched isomers, were tested for feeding deterrent activity against Myzus persicae. The effect of these compounds on probing, feeding and settling behavior of M. persicae was studied in vivo. The deterrent activity of decalactones (4a and 4b against aphids depended on the size of the lactone ring and the enantiomeric purity of the compounds. δ-Decalactone (4b appeared inactive against M. persicae while γ-decalactone (4a restrained aphid probing at ingestional phase. Only (--(S-γ-decalactone (4a had strong and durable (i.e. lasting for at least 24 hours limiting effect, expressed at phloem level.

  7. Effect of permethrin, anthracene and mixture exposure on shell components, enzymatic activities and proteins status in the Mediterranean clam Venerupis decussata.

    Science.gov (United States)

    Sellami, Badreddine; Khazri, Abdelhafidh; Mezni, Amine; Louati, Héla; Dellali, Mohamed; Aissa, Patricia; Mahmoudi, Ezzeddine; Beyrem, Hamouda; Sheehan, David

    2015-01-01

    Anthracene (ANT) and permethrin (PER) are two of the more toxic compounds reaching the marine environment. This study aimed to determine the impact of these molecules on Venerupis decussata, an economically important species cultured on the Tunisian coast. Shell structure and its possible transformation upon exposure to the two contaminants were studied by X-ray diffraction and gravimetric analyses. Results revealed a phase transition in shell composition from aragonite to calcite after PER exposure, to a mixture of PER and ANT (Mix) but not for ANT alone. Catalase (CAT), superoxide dismutase (SOD) and glutathione transferase (GST) activities were determined in digestive gland and gills after exposure to ANT, PER and Mix to assess the impact of the contamination on the oxidative status of V. decussata. Enzyme activities increased in the digestive gland after PER treatment and in the gills after ANT treatment. PER exposure significantly reduced the levels of free thiols and increased levels of carbonylated proteins in the digestive gland, as compared to controls. In contrast, ANT exposure significantly reduced free thiols and increased the number of carbonylated proteins in the gills. Mix induced additive effects as measured by both enzymatic and proteomic approaches. The present study suggests that PER has a strong effect on shell structure; that PER and ANT exposure generate compound-dependent oxidative stress in the tissues of V. decussata and that a mixture of the two compounds has synergistic effects on biochemical response. PMID:25461742

  8. Mechanistic and Structural Studies of Protein-Only RNase P Compared to Ribonucleoproteins Reveal the Two Faces of the Same Enzymatic Activity

    Directory of Open Access Journals (Sweden)

    Cédric Schelcher

    2016-06-01

    Full Text Available RNase P, the essential activity that performs the 5′ maturation of tRNA precursors, can be achieved either by ribonucleoproteins containing a ribozyme present in the three domains of life or by protein-only enzymes called protein-only RNase P (PRORP that occur in eukaryote nuclei and organelles. A fast growing list of studies has investigated three-dimensional structures and mode of action of PRORP proteins. Results suggest that similar to ribozymes, PRORP proteins have two main domains. A clear functional analogy can be drawn between the specificity domain of the RNase P ribozyme and PRORP pentatricopeptide repeat domain, and between the ribozyme catalytic domain and PRORP N4BP1, YacP-like Nuclease domain. Moreover, both types of enzymes appear to dock with the acceptor arm of tRNA precursors and make specific contacts with the corner of pre-tRNAs. While some clear differences can still be delineated between PRORP and ribonucleoprotein (RNP RNase P, the two types of enzymes seem to use, fundamentally, the same catalytic mechanism involving two metal ions. The occurrence of PRORP and RNP RNase P represents a remarkable example of convergent evolution. It might be the unique witness of an ongoing replacement of catalytic RNAs by proteins for enzymatic activities.

  9. Influence of Tableting on Enzymatic Activity of Papain along with Determination of Its Percolation Threshold with Microcrystalline Cellulose

    OpenAIRE

    Sharma, Manu; Sharma, Vinay; Majumdar, Dipak K.

    2014-01-01

    The binary mixture tablets of papain and microcrystalline cellulose (MCC), dicalcium phosphate dihydrate (DCP), carrageenan, tragacanth, and agar were prepared by direct compression. Carrageenan, tragacanth, and agar provided maximum protection to enzyme activity compared to MCC and DCP. However, stability studies indicated highest loss of enzyme activity with carrageenan, tragacanth, and agar. Therefore, compression behaviour of different binary mixtures of papain with MCC at different compa...

  10. EFFECTS OF ENZYMATIC HYDROLYSIS ON THE ANTIOXIDANT ACTIVITY OF WATER- SOLUBLE ELASTIN EXTRACTED FROM BROILER AND SPENT HEN SKIN

    OpenAIRE

    Mehdi Nadalian; Salma Mohamad Yusop; Abdul Salam Babji; Wan Aida Wan Mustapha; Mohd Azri Azman

    2015-01-01

    Poultry by-products are great economic sources that need to be exploited. Poultry skin could be utilized to extract protein particularly elastin, which is often incorporated in the production of functional food, cosmetic industry or medicine due to its antioxidative properties. In this study, water-soluble elastin was successfully extracted from broiler and spent hen skin and analysed for antioxidant activities including DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS and metal chelating activity...

  11. Effect of ferrous chloride on biogas production and enzymatic activities during anaerobic fermentation of cow dung and Phragmites straw.

    Science.gov (United States)

    Zhang, Huayong; Tian, Yonglan; Wang, Lijun; Mi, Xueyue; Chai, Yang

    2016-06-01

    The effect of ferrous (added as FeCl2) on the anaerobic co-digestion of Phragmites straw and cow dung was studied by investigating the biogas properties, pH values, organic matter degradation (COD) and enzyme activities (cellulase, protease and dehydrogenase) at different stages of mesophilic fermentation. The results showed that Fe(2+) addition increased the cumulative biogas yields by 18.1 % by extending the peak period with high daily biogas yields. Meanwhile, the methane (CH4) contents in the Fe(2+) added groups were generally higher than the control group before the 15th day. The pH values were not significantly impacted by Fe(2+) concentrations during the fermentation process. The COD concentrations, cellulase, protease and dehydrogenase activities varied with the added Fe(2+) concentrations and the stages of the fermentation process. At the beginning stage of fermentation (4th day), Fe(2+) addition increased the biogas production by improving the cellulase and dehydrogenase activities which caused a decline in COD. At the peak stage of fermentation (8th day), Fe(2+) addition enhanced the cellulase and protease activities, and resulted in lower COD contents than the control group. When the biogas yields decreased again (13th day), the COD contents varied similar with the protease and dehydrogenase activities, whilst cellulase activities were not sensitive to Fe(2+) concentrations. At the end of fermentation (26th day), Fe(2+) addition decreased the cellulase activities, led to lower COD contents and finally resulted the lower biogas yields than the control group. Taking the whole fermentation process into account, the promoting effect of Fe(2+) addition on biogas yields was mainly attributed to the extension of the gas production peak stage and the improvement of cellulase activities. PMID:26862032

  12. Antifungal Hydroxy Fatty Acids Produced during Sourdough Fermentation: Microbial and Enzymatic Pathways, and Antifungal Activity in Bread

    OpenAIRE

    Black, Brenna A.; Zannini, Emanuele; Curtis, Jonathan M.; Gänzle, Michael G.

    2013-01-01

    Lactobacilli convert linoleic acid to hydroxy fatty acids; however, this conversion has not been demonstrated in food fermentations and it remains unknown whether hydroxy fatty acids produced by lactobacilli have antifungal activity. This study aimed to determine whether lactobacilli convert linoleic acid to metabolites with antifungal activity and to assess whether this conversion can be employed to delay fungal growth on bread. Aqueous and organic extracts from seven strains of lactobacilli...

  13. Enzymatically Regulated Peptide Pairing and Catalysis for the Bioanalysis of Extracellular Prometastatic Activities of Functionally Linked Enzymes.

    Science.gov (United States)

    Li, Hao; Huang, Yue; Yu, Yue; Li, Tianqi; Li, Genxi; Anzai, Jun-Ichi

    2016-01-01

    Diseases such as cancer arise from systematical reconfiguration of interactions of exceedingly large numbers of proteins in cell signaling. The study of such complicated molecular mechanisms requires multiplexed detection of the inter-connected activities of several proteins in a disease-associated context. However, the existing methods are generally not well-equipped for this kind of application. Here a method for analyzing functionally linked protein activities is developed based on enzyme controlled pairing between complementary peptide helix strands, which simultaneously enables elaborate regulation of catalytic activity of the paired peptides. This method has been used to detect three different types of protein modification enzymes that participate in the modification of extracellular matrix and the formation of invasion front in tumour. In detecting breast cancer tissue samples using this method, up-regulated activity can be observed for two of the assessed enzymes, while the third enzyme is found to have a subtle fluctuation of activity. These results may point to the application of this method in evaluating prometastatic activities of proteins in tumour. PMID:27140831

  14. Enzymatic dyeing of wood

    OpenAIRE

    Zille, Andrea; Paulo, Artur Cavaco

    2005-01-01

    This study reports the “in situ” enzymatic dyeing of pinewood samples using a Trametes villosa laccase, in a batchwise process at low temperature and mild pH. Laccase (EC 1.10.3.2) is a multicopper oxidase, which reduces oxygen to water and simultaneously performs one-electron oxidation of many aromatic substrates such as phenols and aromatic amines. The resulting aryloxy radicals undergo further non-enzymatic reactions forming coloured dimeric, oligomeric and polymeric molecules....

  15. Detection of antibacterial activity of an enzymatic hydrolysate generated by processing rainbow trout by-products with trout pepsin.

    Science.gov (United States)

    Wald, Maleen; Schwarz, Karin; Rehbein, Hartmut; Bußmann, Bettina; Beermann, Christopher

    2016-08-15

    Trout by-product hydrolysates, generated using trout pepsin, were characterized and studied in terms of their antibacterial effects against food contaminants and fish farming pathogens. After a hydrolysis time of 25 min, the hydrolysates demonstrated inhibitory activity against several gram-positive and gram-negative bacteria. The degree of hydrolysis (DH) was found to exert a considerable influence on antibacterial activity, with a significant increase in the observed inhibitory effect at the beginning of hydrolysis. The highest antibacterial activity was obtained at a DH of 30% (enzyme/protein ratio 0.04 U/mg of protein, enzyme activity 6.5 U/mg protein, hydrolysis conditions 37°C, pH 3.0). The highest antibacterial activity detected was against the fish farming bacteria Flavobacterium psychrophilum and Renibacterium salmoninarum, with minimal inhibition concentrations of 2mg/ml and 5mg/ml, respectively. The amino acid determination of the hydrolysate (DH 30%) revealed that lysine, leucine, alanine, arginine, glycine, aspartic acid and glutamic acid residues represented the major amino acids. PMID:27006234

  16. Structural and Enzymatic Characterization of ABgp46, a Novel Phage Endolysin with Broad Anti-Gram-Negative Bacterial Activity

    Science.gov (United States)

    Oliveira, Hugo; Vilas Boas, Diana; Mesnage, Stéphane; Kluskens, Leon D.; Lavigne, Rob; Sillankorva, Sanna; Secundo, Francesco; Azeredo, Joana

    2016-01-01

    The present study demonstrates the antibacterial potential of a phage endolysin against Gram-negative pathogens, particularly against multidrug resistant strains of Acinetobacter baumannii. We have cloned, heterologously expressed and characterized a novel endolysin (ABgp46) from Acinetobacter phage vb_AbaP_CEB1 and tested its antibacterial activity against several multidrug-resistant A. baumannii strains. LC-MS revealed that ABgp46 is an N-acetylmuramidase, that is also active over a broad pH range (4.0–10.0) and temperatures up to 50°C. Interestingly, ABgp46 has intrinsic and specific anti-A. baumannii activity, reducing multidrug resistant strains by up to 2 logs within 2 h. By combining ABgp46 with several organic acids that act as outer membrane permeabilizing agents, it is possible to increase and broaden antibacterial activity to include other Gram-negative bacterial pathogens. In the presence of citric and malic acid, ABgp46 reduces A. baumannii below the detection limit (>5 log) and more than 4 logs Pseudomonas aeruginosa and Salmonella typhimurium strains. Overall, this globular endolysin exhibits a broad and high activity against Gram-negative pathogens, that can be enhanced in presence of citric and malic acid, and be used in human and veterinary medicine. PMID:26955368

  17. EFFECTS OF ENZYMATIC HYDROLYSIS ON THE ANTIOXIDANT ACTIVITY OF WATER- SOLUBLE ELASTIN EXTRACTED FROM BROILER AND SPENT HEN SKIN

    Directory of Open Access Journals (Sweden)

    Mehdi Nadalian

    2015-12-01

    Full Text Available Poultry by-products are great economic sources that need to be exploited. Poultry skin could be utilized to extract protein particularly elastin, which is often incorporated in the production of functional food, cosmetic industry or medicine due to its antioxidative properties. In this study, water-soluble elastin was successfully extracted from broiler and spent hen skin and analysed for antioxidant activities including DPPH (1,1-diphenyl-2-picryl hydrazyl, ABTS and metal chelating activity. Antioxidant activity of elastin extracted from broiler skin hydrolysed by Alcalase (EBA and Elastase (EBE also elastin extracted from spent hen skin hydrolyzed by Alcalase (ESA and Elastease (ESE. The EBE, EBA, ESE and ESA had higher DPPH (16-30, 19-35, 29-48 and 31-50%, respectively, ABTS activity ( 73-79, 60-79, 67-79 and 72-79 %, respectively, and Fe2+chelating activity ( 65-69, 50-56, 71-77 and 62-70 %, respectively. This concluded that water-soluble elastin is a bioactive component that could potentially be used in the formulation of functional foods, nutraceuticals, cosmetic and pharmaceutical industry

  18. ENZYMATIC ACTIVITY AND ANTIBIOTIC RESISTANCE PROFILE OF LACTOBACILLUS PARACASEI SSP. PARACASEI-1 ISOLATED FROM REGIONAL YOGURTS OF BANGLADESH

    Directory of Open Access Journals (Sweden)

    Ummay Honi

    2013-12-01

    Full Text Available Lactobacillus paracasei ssp. paracasei-1 was identified from traditional yogurts of Khulna region, Bangladesh and its enzyme and antibiotic resistance profiles were determined. A commercially available API Zym kit was employed to determine the activities of 19 different enzymes. We found that L. paracasei ssp. paracasei-1 showed strong activities for several enzymes, viz. leucine arylamidase, valine arylamidase, napthol-AS-BI-phosphohydrolase, β-galactosidase, α –Glucosidase, N-Acetyl- β- glucosaminidase while activities for other enzymes were absent. Antibiotic resistance profile was assessed by minimum inhibitory concentration (MIC test for 61 major antibiotics and 4 antifungal agents obtained from commercial sources in MRS Agar media. The strain generally showed resistance to gram negative spectrum antibiotic while it showed susceptibility towards β-lactam antibiotic to gram positive spectrum antibiotic. The findings provide the therapeutic basis of using L. paracasei ssp. paracasei-1 in finished food products.

  19. Direct Imaging by Cryo-TEM Shows Membrane Break-up by Phospholipase A2 Enzymatic Activity

    DEFF Research Database (Denmark)

    Callisen, Thomas Hønger; Talmon, Y.

    1998-01-01

    -burst phenomenon reflects a cascade process. The PLA(2)-induced changes in lipid composition transform the morphology which in turn results in an acceleration of the rate of hydrolysis because of a strong coupling between the PLA(2) activity and the morphology of the lipid suspension....... hydrolysis of phospholipid vesicles with respect to changes in lipid composition and morphology. Our direct experimental results show that the initial reaction conditions are strongly perturbed during the course of hydrolysis, Most strikingly, cryo-TEM reveals that starting in the lag phase, vesicles become...... perforated and degrade into open vesicles, bilayer fragments, and micelles, This structural instability extends throughout the system in the activity burst regime. In agreement with earlier reported correlations between initial phospholipase activity and substrate morphology, our results suggest that the lag...

  20. Effect of limited proteolysis on the stability and enzymatic activity of human placental S-adenosylhomocysteine hydrolase.

    OpenAIRE

    Huang, H; Yuan, C. S.; Borchardt, R. T.

    1997-01-01

    Human placental S-adenosylhomocysteine (AdoHcy) hydrolase was subjected to limited papain digestion. The multiple cleavage sites in the enzyme were identified to be Lys94-Ala95, Tyr100-Ala101, Glu243-Ile244, Met367-Ala368, Gln369-Ile370, and Gly382-Val383. Despite multiple cleavage sites in the backbone of the protein, the digested enzyme was able to maintain its quaternary structure and retain its full catalytic activity. The enzyme activity of the partially digested AdoHcy hydrolase was ess...

  1. Small Angle Neutron Scattering Reveals pH-dependent Conformational Changes in Trichoderma reesei Cellobiohydrolase I: IMPLICATIONS FOR ENZYMATIC ACTIVITY*

    OpenAIRE

    Pingali, Sai Venkatesh; O'Neill, Hugh M.; McGaughey, Joseph; Urban, Volker S.; Rempe, Caroline S.; Petridis, Loukas; Jeremy C Smith; Evans, Barbara R.; Heller, William T.

    2011-01-01

    Cellobiohydrolase I (Cel7A) of the fungus Trichoderma reesei (now classified as an anamorph of Hypocrea jecorina) hydrolyzes crystalline cellulose to soluble sugars, making it of key interest for producing fermentable sugars from biomass for biofuel production. The activity of the enzyme is pH-dependent, with its highest activity occurring at pH 4–5. To probe the response of the solution structure of Cel7A to changes in pH, we measured small angle neutron scattering of it in a series of solut...

  2. Comparison of the role that entropy has played in processes of non-enzymatic and enzymatic catalysis

    International Nuclear Information System (INIS)

    The function that entropy has played is compared in processes of non-enzymatic and enzymatic catalysis. The processes followed are showed: the kinetics of the acid hydrolysis of 3-pentyl acetate and cyclopentyl acetate catalyzed by hydrochloric acid and enzymatic hydrolysis of ethyl acetate and γ-butyrolactone catalyzed by pig liver esterase. The activation parameters of Eyring were determined for each process and interpreted the contribution of the entropy of activation for catalysis in this type of model reactions. (author)

  3. Heat treatment of curdlan enhances the enzymatic production of biologically active β-(1,3)-glucan oligosaccharides.

    Science.gov (United States)

    Kumagai, Yuya; Okuyama, Masayuki; Kimura, Atsuo

    2016-08-01

    Biologically active β-(1,3)-glucan oligosaccharides were prepared from curdlan using GH64 enzyme (KfGH64). KfGH64 showed low activity toward native curdlan; thereby pretreatment conditions of curdlan were evaluated. KfGH64 showed the highest activity toward curdlan with heat treatment. The most efficient pretreatment (90°C for 0.5h) converted approximately 60% of curdlan into soluble saccharides under the optimized enzyme reaction conditions (pH 5.5, 37°C, 100rpm mixing speed, 24h, and 10μg of KfGH64/1g of curdlan). The resulting products were predominantly laminaripentaose and a small amount of β-(1,3)-glucans with an average degree of polymerization (DP) of 13 and 130. The products did not contain small oligosaccharides (DPhydrolysis of heat-treated curdlan by KfGH64 is a suitable method for the production of biologically active β-(1,3)-glucan oligosaccharides. PMID:27112889

  4. Enzymatic activity of α-amylase in alimentary tract Spodoptera littoralis (Boisduval (Lepidoptera: Noctuidae: Characterization and Compartmentalization

    Directory of Open Access Journals (Sweden)

    Ali Darvishzadeh

    2014-09-01

    Full Text Available The Egyptian cotton leafworm, Spodoptera littoralis (Boisduval (Lepidoptera: Noctuidae damages a wide variety of crops in Middle East. Their hosts include cotton, alfalfa, eggplant, tomato, lettuce, bean and some ornamental crops. The intensive use of broad-spectrum insecticides against S. littoralis has led to the development of resistance to many registered pesticides use for its control. The purpose of the present study is biochemical characterization of digestive enzymes of this pest to gain a better understanding of the digestive physiology. The physiology and biochemistry of the insect digestive enzyme had an important role in the study of novel insecticidal strategies. The Egyptian cotton leafworm alimentary canal consists of a short foregut, a long midgut and a short hindgut. Application of pH indicators showed that alimentary canal was alkaline. Our results showed that activities of gut α-amylase were different in three parts of the insect gut. Also shown the greatest activity of α-amylase observed in the midgut followed by hindgut and foregut, respectively. However, there were not significant differences in activity of the enzyme in the midgut and hindgut. The optimal pH α-amylase in foregut, midgut and hindgut were 10.0. Zymogram analysis of different part of gut showed four bands in midgut, hind gut and two bands in foregut. Therefore, in midgut of S. littoralis, four isoenzymes were present. These results explain why more amylase activity was seen in these regions in the spectrophotometric assay.

  5. Enzymatic Synthesis of Structured Lipids using a Novel Cold-Active Lipase from Pichia lynferdii NRRL Y-7723

    Science.gov (United States)

    Structured lipids (SL) were synthesized by the acidolysis of borage oil with caprylic acid using lipases. Six commercial lipases from different sources and a novel lipase from Pichia lynferdii NRRL Y-7723 were screened for their acidolysis activities and Lipozyme RM IM and NRRL Y-7723 lipase were s...

  6. Effect of salinity on biomass production and activities of some key enzymatic antioxidants in kochia (kochia scoparia)

    International Nuclear Information System (INIS)

    Soil salinity is a major constraint to food production due to its negative impact on crop yield. Kochia (Kochia scoparia) is a salinity-resistant plant that can widely be used as emergency forage for livestock by using saline waters and soils in desert ecosystems. In order to investigate physiological mechanism, antioxidants activity and potential production of Kochia in response to different levels of salinity, an experiment was performed in a split plot based on randomized complete block design with three replications. Saline waters (5.2, 10.5 and 23.1 dS m/sup -1/) and three Kochia ecotypes (Birjand, Borujerd and Sabzevar) were allocated as main and sub plots, respectively. The results showed that salinity did not impose any significant effect on dry matter production but relative water content (RWC) and seed yield decreased by salinity stress. In general, no positive correlation coefficient was observed between dry matter production and physiological and biochemical parameters except superoxide dismutase (SOD) at 23.1 dS m/sup -1/. There was no significant difference among ecotypes in dry matter production and seed yield. Sabzevar ecotype showed the highest proline, total phenol content and peroxidase (POX) activity. Ascorbate peroxidase (APX), catalase (CTA), and superoxide dismutase (SOD) activity was higher in Borujerd ecotype, while highest soluble sugar, glutathione reductase (GR) activity and DPPH - radical scavenging activity was observed in Birjand ecotype. According to these results, Kochia has a reliable tolerance to elevated levels of salinities up to 23 dS m/sup -1/ and it seems that it can control oxidative stress by continuing growth. (author)

  7. Diversity and extracellular enzymatic activities of yeasts isolated from King George Island, the sub-Antarctic region

    Directory of Open Access Journals (Sweden)

    Carrasco Mario

    2012-11-01

    Full Text Available Abstract Background Antarctica has been successfully colonized by microorganisms despite presenting adverse conditions for life such as low temperatures, high solar radiation, low nutrient availability and dryness. Although these “cold-loving” microorganisms are recognized as primarily responsible for nutrient and organic matter recycling/mineralization, the yeasts, in particular, remain poorly characterized and understood. The aim of this work was to study the yeast microbiota in soil and water samples collected on King George Island. Results A high number of yeast isolates was obtained from 34 soil and 14 water samples. Molecular analyses based on rDNA sequences revealed 22 yeast species belonging to 12 genera, with Mrakia and Cryptococcus genera containing the highest species diversity. The species Sporidiobolus salmonicolor was by far the most ubiquitous, being identified in 24 isolates from 13 different samples. Most of the yeasts were psychrotolerant and ranged widely in their ability to assimilate carbon sources (consuming from 1 to 27 of the 29 carbon sources tested. All species displayed at least 1 of the 8 extracellular enzyme activities tested. Lipase, amylase and esterase activity dominated, while chitinase and xylanase were less common. Two yeasts identified as Leuconeurospora sp. and Dioszegia fristingensis displayed 6 enzyme activities. Conclusions A high diversity of yeasts was isolated in this work including undescribed species and species not previously isolated from the Antarctic region, including Wickerhamomyces anomalus, which has not been isolated from cold regions in general. The diversity of extracellular enzyme activities, and hence the variety of compounds that the yeasts may degrade or transform, suggests an important nutrient recycling role of microorganisms in this region. These yeasts are of potential use in industrial applications requiring high enzyme activities at low temperatures.

  8. A biodegradation study of forest biomass by Aspergillus niger F7: correlation between enzymatic activity, hydrolytic percentage and biodegradation index

    Directory of Open Access Journals (Sweden)

    Nivedita Sharma

    2012-06-01

    Full Text Available Aspergillus niger F7 isolated from soil was found to be the potent producer of cellulase and xylanase. The residue of forest species Toona ciliata, Celtris australis, Cedrus deodara and Pinus roxburghii was selected as substrate for biodegradation study due to its easy availability and wide use in industry. It was subjected to alkali (sodium hydroxide treatment for enhancing its degradation. Biodegradation of forest waste by hydrolytic enzymes (cellulase and xylanase secreted by A. niger under solid state fermentation (SSF was explored. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Highest extracellular enzyme activity of 2201±23.91 U/g by A. niger was shown in pretreated C. australis wood resulting in 6.72±0.20 percent hydrolysis and 6.99±0.23 biodegradation index (BI. The lowest BI of 1.40±0.08 was observed in untreated saw dust of C. deodara having the least enzyme activity of 238±1.36 U/g of dry matter. Biodegradation of forest biomass under SSF was increased many folds when moistening agent i.e. tap water had been replaced with modified basal salt media (BSM. In BSM mediated degradation of forest waste with A. niger, extracellular enzyme activity was increased up to 4089±67.11 U/g of dry matter in turn resulting in higher BI of 15.4±0.41 and percent hydrolysis of 19.38±0.81 in pretreated C. australis wood. A. niger exhibited higher enzyme activity on pretreated biomass when moistened with modified BSM in this study. Statistically a positive correlation has been drawn between these three factors i.e. enzyme activity, BI and percent hydrolysis of forest biomass thus proving their direct relationship with each other.

  9. Data of enzymatic activities of the electron transport chain and ATP synthase complexes in mouse hepatoma cells following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

    Science.gov (United States)

    Hwang, Hye Jin; Steidemann, Michelle; Dunivin, Taylor K; Rizzo, Mike; LaPres, John J

    2016-09-01

    2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the most widely studied ligand of the aryl hydrocarbon receptor (AHR). The AHR-dependent TCDD-induced mitochondrial hyperpolarization (Tappenden et al., 2011) [1] and reduced oxygen consumption rate of intact mouse hepatoma cells (Huang et al., in press) [2] in the previous studies suggest that these alterations can be related to enzymatic activities of the electron transport chain (ETC) and ATP synthase in oxidative phosphorylation (OXPHOS) system. Here, we evaluated the activity of each complex in the OXPHOS system using in vitro enzymatic assays. The calculated enzymatic activity of each complex was normalized against the activity of citrate synthase. To combine each value from an independent experiment, normalized enzyme activities from cells exposed to TCDD were converted to fold changes via comparison to the activity relative to time-matched vehicle control. The averaged fold change for each treatment suggests more replicates are needed in order to clearly evaluate a difference between treatments. PMID:27284569

  10. Comparative study of human intestinal and hepatic esterases as related to enzymatic properties and hydrolizing activity for ester-type drugs.

    Science.gov (United States)

    Inoue, M; Morikawa, M; Tsuboi, M; Ito, Y; Sugiura, M

    1980-08-01

    In attempts to determine the exact role of intestinal esterase in the body, we purified esterases from human intestinal mucosa and liver, and compared the enzymatic properties and substrate specificities with those of purified esterases. Esterase from human liver was purified 58-fold, by treatment with butanol, DE-52 and DEAE Sephadex A-50 column chromatographies, Sephadex G-200 gel filtration, and isoelectric focusing. The purified preparation showed a single band by polyacylamide gel electrophoresis. The molecular weights of intestinal and hepatic esterases were determined to be 53,000-55,000 and 180,000, respectively, by gel filtration on Sephadex G-200. The activity of the purified intestinal and hepatic esterases was strongly inhibited by diethyl-p-nitrophenyl phosphate and diisopropyl fluorophosphate, and was not inhibited by eserine sulfate and p-chloromercuribenzoate. Moreover, the purified esterases hydrolyzed ester-type drugs such as aspirin, clofibrate, indanyl carbenicillin and procaine. Hepatic esterase had properties similar to those of intestinal esterase with respect to the sensitivity to organophosphate and the substrate specificity. However, the two purified esterases differed in properties such as molecular weight, isoelectric point, thermostability and optimal pH. PMID:7206363

  11. Microwave-assisted aqueous enzymatic extraction of oil from pumpkin seeds and evaluation of its physicochemical properties, fatty acid compositions and antioxidant activities.

    Science.gov (United States)

    Jiao, Jiao; Li, Zhu-Gang; Gai, Qing-Yan; Li, Xiao-Juan; Wei, Fu-Yao; Fu, Yu-Jie; Ma, Wei

    2014-03-15

    Microwave-assisted aqueous enzymatic extraction (MAAEE) of pumpkin seed oil was performed in this study. An enzyme cocktail comprised of cellulase, pectinase and proteinase (w/w/w) was found to be the most effective in releasing oils. The highest oil recovery of 64.17% was achieved under optimal conditions of enzyme concentration (1.4%, w/w), temperature (44°C), time (66 min) and irradiation power (419W). Moreover, there were no significant variations in physicochemical properties of MAAEE-extracted oil (MAAEEO) and Soxhlet-extracted oil (SEO), but MAAEEO exhibited better oxidation stability. Additionally, MAAEEO had a higher content of linoleic acid (57.33%) than SEO (53.72%), and it showed stronger antioxidant activities with the IC50 values 123.93 and 152.84, mg/mL, according to DPPH radical scavenging assay and β-carotene/linoleic acid bleaching test. SEM results illustrated the destruction of cell walls and membranes by MAAEE. MAAEE is, therefore, a promising and environmental-friendly technique for oil extraction in the food industry. PMID:24206680

  12. Functional significance of Asn-linked glycosylation of proteinase 3 for enzymatic activity, processing, targeting, and recognition by anti-neutrophil cytoplasmic antibodies.

    Science.gov (United States)

    Specks, Ulrich; Fass, David N; Finkielman, Javier D; Hummel, Amber M; Viss, Margaret A; Litwiller, Robert D; McDonald, Cari J

    2007-01-01

    Proteinase 3 (PR3) is a neutral serine protease stored in neutrophil granules. It has substantial sequence homology with elastase, cathepsin G and azurocidin. PR3 is the target antigen for autoantibodies (ANCA) in Wegener's granulomatosis, a necrotizing vasculitis syndrome. ANCA have been implicated in the pathogenesis of this disease. PR3 has two potential Asn-linked glycosylation sites. This study was designed to determine the occupancy of these glycosylation sites, and to evaluate their effect on enzymatic function, intracellular processing, targeting to granules and recognition by ANCA. We found that glycosylation occurs at both sites in native neutrophil PR3 and in wild type recombinant PR3 (rPR3) expressed in HMC-1 cells. Using glycosylation deficient rPR3 mutants we found that glycosylation at Asn-147, but not at Asn-102, is critical for thermal stability, and for optimal hydrolytic activity of PR3. Efficient amino-terminal proteolytic processing of rPR3 is dependent on glycosylation at Asn-102. Targeting to granules is not dependent on glycosylation, but unglycosylated rPR3 gets secreted preferentially into media supernatants. Finally, a capture ELISA for ANCA detection, using rPR3 glycosylation variants as target antigens, reveals that in about 20% of patients, epitope recognition by ANCA is affected by the glycosylation status of PR3. PMID:17158864

  13. Alterations in certain Enzymatic Activities in Liver and Serum of Male Rats Treated with Endotoxin Or Exposed To Gamma Radiation

    International Nuclear Information System (INIS)

    This study was performed to determine the effects of endotoxins and gamma radiation on glutamic oxalacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), acid phosphatase (ACP) and alkaline phosphatase (ALP). Endotoxin (isolated from Escherichia coli, Serotype 055) was supplemented to rats by gavage with a dose level of 20 mg/kg /day for a period of four weeks. Whole body gamma irradiation was carried out by exposure of rats to 4 Gy delivered as 0.5 Gy twice weekly. The results demonstrated that endotoxin administration as well as exposure to gamma radiation produced significant increases in the activities of liver transaminases and acid phosphatase enzymes 1,2 and 4 weeks post treatment. In the serum of endotoxin treated rats, the activities of transaminases showed non-significant changes while significant increases were observed in the serum of irradiated rats

  14. An enzymatic deconjugation method for the analysis of small molecule active drugs on antibody-drug conjugates.

    Science.gov (United States)

    Li, Yi; Gu, Christine; Gruenhagen, Jason; Yehl, Peter; Chetwyn, Nik P; Medley, Colin D

    2016-01-01

    Antibody-drug conjugates (ADCs) are complex therapeutic agents that use the specific targeting properties of antibodies and the highly potent cytotoxicity of small molecule drugs to selectively eliminate tumor cells while limiting the toxicity to normal healthy tissues. Two critical quality attributes of ADCs are the purity and stability of the active small molecule drug linked to the ADC, but these are difficult to assess once the drug is conjugated to the antibody. In this study, we report a enzyme deconjugation approach to cleave small molecule drugs from ADCs, which allows the drugs to be subsequently characterized by reversed-phase high performance liquid chromatography. The model ADC we used in this study utilizes a valine-citrulline linker that is designed to be sensitive to endoproteases after internalization by tumor cells. We screened several proteases to determine the most effective enzyme. Among the 3 cysteine proteases evaluated, papain had the best efficiency in cleaving the small molecule drug from the model ADC. The deconjugation conditions were further optimized to achieve complete cleavage of the small molecule drug. This papain deconjugation approach demonstrated excellent specificity and precision. The purity and stability of the active drug on an ADC drug product was evaluated and the major degradation products of the active drug were identified. The papain deconjugation method was also applied to several other ADCs, with the results suggesting it could be applied generally to ADCs containing a valine-citrulline linker. Our results indicate that the papain deconjugation method is a powerful tool for characterizing the active small molecule drug conjugated to an ADC, and may be useful in ensuring the product quality, efficacy and the safety of ADCs. PMID:26891281

  15. Synthesis and enzymatic photo-activity of an O2 tolerant hydrogenase-CdSe@CdS quantum rod bioconjugate.

    Science.gov (United States)

    Hamon, C; Ciaccafava, A; Infossi, P; Puppo, R; Even-Hernandez, P; Lojou, E; Marchi, V

    2014-05-21

    This communication reports on the preparation of stable and photo-active nano-heterostructures composed of O2 tolerant [NiFe] hydrogenase extracted from the Aquifex aeolicus bacterium grafted onto hydrophilic CdSe/CdS quantum rods in view of the development of H2/O2 biofuel cells. The resulting complex is efficient towards H2 oxidation, displays good stability and new photosensitive properties. PMID:24468861

  16. Ureases display biological effects independent of enzymatic activity: Is there a connection to diseases caused by urease-producing bacteria?

    Directory of Open Access Journals (Sweden)

    D. Olivera-Severo

    2006-07-01

    Full Text Available Ureases are enzymes from plants, fungi and bacteria that catalyze the hydrolysis of urea to form ammonia and carbon dioxide. While fungal and plant ureases are homo-oligomers of 90-kDa subunits, bacterial ureases are multimers of two or three subunit complexes. We showed that some isoforms of jack bean urease, canatoxin and the classical urease, bind to glycoconjugates and induce platelet aggregation. Canatoxin also promotes release of histamine from mast cells, insulin from pancreatic cells and neurotransmitters from brain synaptosomes. In vivo it induces rat paw edema and neutrophil chemotaxis. These effects are independent of ureolytic activity and require activation of eicosanoid metabolism and calcium channels. Helicobacter pylori, a Gram-negative bacterium that colonizes the human stomach mucosa, causes gastric ulcers and cancer by a mechanism that is not understood. H. pylori produces factors that damage gastric epithelial cells, such as the vacuolating cytotoxin VacA, the cytotoxin-associated protein CagA, and a urease (up to 10% of bacterial protein that neutralizes the acidic medium permitting its survival in the stomach. H. pylori whole cells or extracts of its water-soluble proteins promote inflammation, activate neutrophils and induce the release of cytokines. In this paper we review data from the literature suggesting that H. pylori urease displays many of the biological activities observed for jack bean ureases and show that bacterial ureases have a secretagogue effect modulated by eicosanoid metabolites through lipoxygenase pathways. These findings could be relevant to the elucidation of the role of urease in the pathogenesis of the gastrointestinal disease caused by H. pylori.

  17. Molecular cloning and expression of gene fragments from corynebacteriophage beta encoding enzymatically active peptides of diphtheria toxin.

    OpenAIRE

    Tweten, R K; Collier, R J

    1983-01-01

    Two restriction fragments from corynebacteriophage beta vir tox+ that encode peptides similar to diphtheria toxin fragment A and the chain termination fragment, CRM45, have been cloned into Escherichia coli in plasmid pBR322. Clones containing the recombinant plasmids produced gene products that were active in catalyzing the ADP ribosylation of elongation factor 2 and were reactive with diphtheria toxin antiserum. Toxin-related peptides were found primarily in the periplasmic compartment and ...

  18. Evaluation of oil removal efficiency and enzymatic activity in some fungal strains for bioremediation of petroleum-polluted soils

    OpenAIRE

    Fariba Mohsenzadeh; Abdolkarim Chehregani Rad; Mehrangiz Akbari

    2012-01-01

    Abstract Background Petroleum pollution is a global disaster and there are several soil cleaning methods including bioremediation. Methods In a field study, fugal strains were isolated from oil-contaminated sites of Arak refinery (Iran) and their growth ability was checked in potato dextrose agar (PDA) media containing 0-10% v/v crude oil, the activity of three enzymes (Catalase, Peroxidase and Phenol Oxidase) was evaluated in the fungal colonies and bioremediation ability of the fungi was ch...

  19. Comparative Composition and Antioxidant Activity of Peptide Fractions Obtained by Ultrafiltration of Egg Yolk Protein Enzymatic Hydrolysates

    Directory of Open Access Journals (Sweden)

    Yves Pouliot

    2011-07-01

    Full Text Available The objective of the study was to compare the antioxidant activity of two distinct hydrolysates and their peptide fractions prepared by ultrafiltration (UF using membranes with molecular weight cut-off of 5 and 1 kDa. The hydrolysates were a delipidated egg yolk protein concentrate (EYP intensively hydrolyzed with a combination of two bacterial proteases, and a phosphoproteins (PPP extract partially hydrolyzed with trypsin. Antioxidant activity, as determined by the oxygen radical absorbance capacity (ORAC assay, was low for EYP and PPP hydrolysates with values of 613.1 and 489.2 µM TE×g−1 protein, respectively. UF-fractionation of EYP hydrolysate increased slightly the antioxidant activity in permeate fractions (720.5–867.8 µM TE×g−1 protein. However, ORAC values were increased by more than 3-fold in UF-fractions prepared from PPP hydrolysate, which were enriched in peptides with molecular weight lower than 5 kDa. These UF-fractions were characterized by their lower N/P atomic ratio and higher phosphorus content compared to the same UF-fractions obtained from EYP-TH. They also contained high amounts of His, Met, Leu, and Phe, which are recognized as antioxidant amino acids, but also high content in Lys and Arg which both represent target amino acids of trypsin used for the hydrolysis of PPP.

  20. Curcumin Blocks Naproxen-Induced Gastric Antral Ulcerations through Inhibition of Lipid Peroxidation and Activation of Enzymatic Scavengers in Rats.

    Science.gov (United States)

    Kim, Jeong-Hwan; Jin, Soojung; Kwon, Hyun Ju; Kim, Byung Woo

    2016-08-28

    Curcumin is a polyphenol derived from the plant Curcuma longa, which is used for the treatment of diseases associated with oxidative stress and inflammation. The present study was undertaken to determine the protective effect of curcumin against naproxen-induced gastric antral ulcerations in rats. Different doses (10, 50, and 100 mg/kg) of curcumin or vehicle (curcumin, 0 mg/kg) were pretreated for 3 days by oral gavage, and then gastric mucosal lesions were caused by 80 mg/kg naproxen applied for 3 days. Curcumin significantly inhibited the naproxen-induced gastric antral ulcer area and lipid peroxidation in a dose-dependent manner. In addition, curcumin markedly increased activities of radical scavenging enzymes, such as superoxide dismutase (SOD), catalase, and glutathione peroxidase in a dose-dependent manner. Specifically, 100 mg/kg curcumin completely protected the gastric mucosa against the loss in the enzyme, resulting in a drastic increase of activities of radical scavenging enzymes up to more than the level of untreated normal rats. Histological examination obviously showed that curcumin prevents naproxen-induced gastric antral ulceration as a result of direct protection of the gastric mucosa. These results suggest that curcumin blocks naproxen-induced gastric antral ulcerations through prevention of lipid peroxidation and activation of radical scavenging enzymes, and it may offer a potential remedy of gastric antral ulcerations. PMID:27197667

  1. Enzymatic Synthesis of l-Ascorbyl Fatty Acid Esters Under Ultrasonic Irradiation and Comparison of Their Antioxidant Activity and Stability.

    Science.gov (United States)

    Jiang, Chen; Lu, Yuyun; Li, Zhuo; Li, Cunzhi; Yan, Rian

    2016-06-01

    A series of novel l-ascorbyl fatty acid esters were synthesized by catalization of Novozym(®) 435 under ultrasonic irradiation and characterized by infrared spectroscopy, electrospray ionization mass spectra, and nuclear magnetic resonance. Their properties especially antioxidant activity and stability were investigated. The results showed that the reducing power, the scavenging activity of hydroxyl radical and 2,2-diphenyl-1-picrylhydrazyl radical were decreased with the increase of the number of carbon atoms in fatty acid. The hydroxyl radical scavenging activity and reducing power of l-ascorbyl saturated fatty acid esters were better than that of tert-butylhydroquinone. The induction period in lipid oxidation of l-ascorbyl saturated fatty acid esters and tert-butylhydroquinone were longer than that of l-ascorbyl unsaturated fatty acid esters and l-ascorbic acid both in soybean oil and lard. Besides, the l-ascorbyl fatty acid esters showed different stabilities in different conditions by comparing with l-ascorbic acid, and the l-ascorbyl saturated fatty acid esters were more stable than l-ascorbyl unsaturated fatty acid esters in ethanol solution. PMID:27100741

  2. Structural basis of enzymatic activity for the ferulic acid decarboxylase (FADase from Enterobacter sp. Px6-4.

    Directory of Open Access Journals (Sweden)

    Wen Gu

    Full Text Available Microbial ferulic acid decarboxylase (FADase catalyzes the transformation of ferulic acid to 4-hydroxy-3-methoxystyrene (4-vinylguaiacol via non-oxidative decarboxylation. Here we report the crystal structures of the Enterobacter sp. Px6-4 FADase and the enzyme in complex with substrate analogues. Our analyses revealed that FADase possessed a half-opened bottom β-barrel with the catalytic pocket located between the middle of the core β-barrel and the helical bottom. Its structure shared a high degree of similarity with members of the phenolic acid decarboxylase (PAD superfamily. Structural analysis revealed that FADase catalyzed reactions by an "open-closed" mechanism involving a pocket of 8 × 8 × 15 Å dimension on the surface of the enzyme. The active pocket could directly contact the solvent and allow the substrate to enter when induced by substrate analogues. Site-directed mutagenesis showed that the E134A mutation decreased the enzyme activity by more than 60%, and Y21A and Y27A mutations abolished the enzyme activity completely. The combined structural and mutagenesis results suggest that during decarboxylation of ferulic acid by FADase, Trp25 and Tyr27 are required for the entering and proper orientation of the substrate while Glu134 and Asn23 participate in proton transfer.

  3. Role of enzymatic activity in muscle damage and cytotoxicity induced by Bothrops asper Asp49 phospholipase A2 myotoxins: are there additional effector mechanisms involved?

    Directory of Open Access Journals (Sweden)

    Diana Mora-Obando

    2014-09-01

    Full Text Available Viperid venoms often contain mixtures of Asp49 and Lys49 PLA2 myotoxin isoforms, relevant to development of myonecrosis. Given their difference in catalytic activity, mechanistic studies on each type require highly purified samples. Studies on Asp49 PLA2s have shown that enzyme inactivation using p-bromophenacyl bromide (p-BPB drastically affects toxicity. However, based on the variable levels of residual toxicity observed in some studies, it has been suggested that effector mechanisms independent of catalysis may additionally be involved in the toxicity of these enzymes, possibly resembling those of the enzymatically inactive Lys49 myotoxins. A possibility that Lys49 isoforms could be present in Asp49 PLA2 preparations exists and, if undetected in previous studies, could explain the variable residual toxicity. This question is here addressed by using an enzyme preparation ascertained to be free of Lys49 myotoxins. In agreement with previous reports, inactivation of the catalytic activity of an Asp49 myotoxin preparation led to major inhibition of toxic effects in vitro and in vivo. The very low residual levels of myotoxicity (7% and cytotoxicity (4% observed can be attributed to the low, although detectable, enzyme remaining active after p-BPB treatment (2.7%, and would be difficult to reconcile with the proposed existence of additional catalytic-independent toxic mechanisms. These findings favor the concept that the effector mechanism of toxicity of Asp49 PLA2 myotoxins from viperids fundamentally relies on their ability to hydrolyze phospholipids, arguing against the proposal that membrane disruption may also be caused by additional mechanisms that are independent of catalysis.

  4. Enzymatic removal of estrogenic activity of nonylphenol and octylphenol aqueous solutions by immobilized laccase from Trametes versicolor

    Energy Technology Data Exchange (ETDEWEB)

    Catapane, Maria [Institute of Genetics and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples (Italy); National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d’Oro, 305, 00136 Rome (Italy); Nicolucci, Carla; Menale, Ciro; Mita, Luigi [National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d’Oro, 305, 00136 Rome (Italy); Department of Experimental Medicine, Second University of Naples, Via S. M. di Costantinopoli, 16, 80138 Naples (Italy); Rossi, Sergio [Institute of Genetics and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples (Italy); Mita, Damiano G., E-mail: mita@igb.cnr.it [Institute of Genetics and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples (Italy); National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d’Oro, 305, 00136 Rome (Italy); Department of Experimental Medicine, Second University of Naples, Via S. M. di Costantinopoli, 16, 80138 Naples (Italy); Diano, Nadia [Institute of Genetics and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples (Italy); National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d’Oro, 305, 00136 Rome (Italy); Department of Experimental Medicine, Second University of Naples, Via S. M. di Costantinopoli, 16, 80138 Naples (Italy)

    2013-03-15

    Highlights: ► Endocrine disruptors cause adverse effects in living organisms. ► Nonylphenol and Octylphenol are alkylphenols recognized as endocrine disruptors. ► It is necessary to remove or reduce their presence in the environment. ► Waters polluted by these pollutants have been bioremediated by immobilized laccase from Trametes versicolor. ► Laccase treated solutions were found to have lost any estrogenic activity. -- Abstract: A fluidized bed reactor, filled with laccase-based beads, has been employed to bioremediate aqueous solutions polluted by endocrine disruptors belonging to the alkylphenols (APs) class. In particular Octylphenol and Nonylphenol have been studied. The catalytic activity of free and immobilized laccase from Trametes versicolor has been characterized as a function of pH, temperature and substrate concentration in the reaction medium. In view of practical applications for each substrate concentration the removal efficiency (RE), the time to halve the initial concentration (τ{sub 50}), and the t{sub c=0}, i.e. the time to reach complete pollutant removal, have been calculated. The immobilized laccase exhibited a lower affinity for octylphenol (K{sub m} = 1.11 mM) than for Nonylphenol (K{sub m} = 0.72 mM), but all the other parameters of applicative interest resulted more significant for octylphenol. For example, the times to reach the complete removal of octylphenol compared to those for nonylphenol at the same concentration is shorter of about 15% (at low concentrations) up to 40% (at high concentrations). The study of cell proliferation with MPP89 cells, a human mesothelioma cell line, and the assay with the YES test indicated the loss of estrogenic activity of the APs solutions after laccase treatment.

  5. High-level expression of enzymatically active mature human gamma-glutamyltransferase in transgenic V79 Chinese hamster cells.

    OpenAIRE

    Visvikis, A; Thioudellet, C; Oster, T; Fournel-Gigleux, S; Wellman, M; Siest, G

    1991-01-01

    gamma-Glutamyltransferase [GGT; (5-glutamyl)-peptide:amino-acid 5-glutamyltransferase, EC 2.3.2.2] is a glutathione-metabolizing enzyme, whose activity variations in serum and organs are valuable markers of preneoplastic processes, alcohol abuse, and induction by xenobiotics. To elucidate the implication of GGT in various metabolic pathways, we established a stable transgenic V79 cell line, highly producing the human GGT. A full-length cDNA, encoding the human hepatoma HepG2 GGT, was subclone...

  6. Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus.

    Science.gov (United States)

    Piazza, Rômi S; Trevisan, Rafael; Flores-Nunes, Fabrício; Toledo-Silva, Guilherme; Wendt, Nestor; Mattos, Jacó J; Lima, Daína; Taniguchi, Satie; Sasaki, Silvio Tarou; Mello, Álvaro C P; Zacchi, Flávia L; Serrano, Miguel A S; Gomes, Carlos H A M; Bícego, Márcia C; Almeida, Eduardo A de; Bainy, Afonso C D

    2016-08-01

    Understanding the mechanism of phenanthrene (PHE) biotransformation and related cellular responses in bivalves can be an important tool to elucidate the risks of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. In the present study it was analyzed the transcriptional levels of 13 biotransformation genes related to cytochrome P450 (CYP), glutathione S-transferase (GST), sulfotransferase (SULT), flavin-containing monooxygenase and fatty acid-binding proteins by qPCR in gill of scallops Nodipecten nodosus exposed for 24 or 96h to 50 or 200μgL(-1) PHE (equivalent to 0.28 and 1.12μM, respectively), followed by depuration in clean water for 96h (DEP). Likewise, it was quantified the activity of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH), GST and levels of lipid peroxidation. Increased transcriptional levels of CYP2UI-like, CYP2D20-like, CYP3A11-like, GSTomega-like, SULT1B1-like genes were detected in organisms exposed to PHE for 24 or 96h. In parallel, GR and GPX activities increased after 96h exposure to 200μgL(-1) PHE and G6PDH activity increased after 24h exposure to 50μgL(-1) PHE. This enhancement of antioxidant and phase I and II biotransformation systems may be related to the 2.7 and 12.5 fold increases in PHE bioaccumulation after 96h exposure to 50 and 200μgL(-1) PHE, respectively. Interestingly, DEP caused reestablishment of GPX and GR activity, as well as to the transcript levels of all upregulated biotransformation genes (except for SULT1B1-like). Bioaccumulated PHE levels decreased 2.5-2.9 fold after depuration, although some biochemical and molecular modifications were still present. Lipid peroxidation levels remained lower in animals exposed to 200μgL(-1) PHE for 24h and DEP. These data indicate that N. nodosus is able to induce an antioxidant and biotransformation-related response to PHE exposure, counteracting its toxicity, and DEP can

  7. Blood Picture and Enzymatic Activities in Common Crap Cyprinus carpio Influenced by Sodium Chloride (NaCl)

    OpenAIRE

    Al-Taee, Shahba Kale Al-Taee; Al-Hamdani, Alaa Hussain

    2014-01-01

    The aim of this study was to assess the effect of Sodium Chloride NaCl on blood picture as hemoglobin (Hb), packed cell volume (PCV) and lymphocyte count also the activity of both enzymes Alanine Amino Transferase (ALT) and Creatine Phospho Kinase (CPK) in Cyprinus carpio. NaCl was used at different concentration (0.5; 1.0 and 1.5 mg\\L) for 7 day. The concentration 1.5 mg\\L was toxic to the fish and caused death after 48 hour of exposure, while in the fish that were exposure separately to bot...

  8. Exotoxin A of Pseudomonas aeruginosa: substitution of glutamic acid 553 with aspartic acid drastically reduces toxicity and enzymatic activity.

    OpenAIRE

    Douglas, C M; Collier, R J

    1987-01-01

    Glutamic acid 553 of Pseudomonas aeruginosa exotoxin A (ETA) has been identified by photoaffinity labeling as a residue within the NAD binding site (S.F. Carroll and R.J. Collier, J. Biol. Chem. 262:8707-8711, 1987). To explore the function of Glu-553 we used oligonucleotide-directed mutagenesis to replace this residue with Asp in cloned ETA and expressed the mutant gene in Escherichia coli K-12. ADP-ribosylation activity of Asp-553 ETA in cell extracts was about 1,800-fold lower and toxicity...

  9. The Nudix Hydrolase CDP-Chase, a CDP-Choline Pyrophosphatase, Is an Asymmetric Dimer with Two Distinct Enzymatic Activities

    Energy Technology Data Exchange (ETDEWEB)

    Duong-Ly, Krisna C.; Gabelli, Sandra B.; Xu, WenLian; Dunn, Christopher A.; Schoeffield, Andrew J.; Bessman, Maurice J.; Amzel, L. Mario (Loyola); (JHU)

    2011-09-06

    A Nudix enzyme from Bacillus cereus catalyzes the hydrolysis of CDP-choline to produce CMP and phosphocholine. Here, we show that in addition, the enzyme has a 3{prime} {yields} 5{prime} RNA exonuclease activity. The structure of the free enzyme, determined to a 1.8-{angstrom} resolution, shows that the enzyme is an asymmetric dimer. Each monomer consists of two domains, an N-terminal helical domain and a C-terminal Nudix domain. The N-terminal domain is placed relative to the C-terminal domain such as to result in an overall asymmetric arrangement with two distinct catalytic sites: one with an 'enclosed' Nudix pyrophosphatase site and the other with a more open, less-defined cavity. Residues that may be important for determining the asymmetry are conserved among a group of uncharacterized Nudix enzymes from Gram-positive bacteria. Our data support a model where CDP-choline hydrolysis is catalyzed by the enclosed Nudix site and RNA exonuclease activity is catalyzed by the open site. CDP-Chase is the first identified member of a novel Nudix family in which structural asymmetry has a profound effect on the recognition of substrates.

  10. DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I

    DEFF Research Database (Denmark)

    Marcussen, Lærke Bay; Jepsen, Morten Leth; Kristoffersen, Emil Laust;

    2013-01-01

    specific for topoisomerase I even in raw cell extracts and presents a simple mean of following enzyme kinetics using standard laboratory equipment such as a qPCR machine or fluorimeter. Human topoisomerase I is a well-known target for the clinically used anti-cancer drugs of the camptothecin family. The...... cytotoxic effect of camptothecins correlates directly with the intracellular topoisomerase I activity. We therefore envision that the presented sensor may find use for the prediction of cellular drug response. Moreover, inhibition of topoisomerase I by camptothecin is readily detectable using the presented...... DNA sensor, suggesting a potential application of the sensor for first line screening for potential topoisomerase I targeting anti-cancer drugs...

  11. Isotopic and enzymatic analyses of planktonic nitrogen utilisation in the vicinity of Cape Sines (Portugal) during weak upwelling activity

    Science.gov (United States)

    Slawyk, Gerd; Coste, Bernard; Collos, Yves; Rodier, Martine

    1997-01-01

    Using measurements of 15N uptake and activities of nitrate reductase and glutamine synthetase, the utilization of nitrogenous nutrients by microplankton in the Portuguese upwelling area was investigated. During this cruise the euphotic zone of coastal waters was in most cases bisected by a nitracline forming two layers. Total inorganic nitrogen uptake rates (NH 4+ + NO 3-) in the upper mixed and nitrate-impoverished layer ranged from 0.1 to 0.8 nM h -1 and were primarily supported by regenerated (ammonium) nitrogen (62-97%), whereas they varied between 0.9 and 10.4 nM h -1 in the deep nitrate-rich layer and were mainly driven by new (nitrate) nitrogen (52-82%). Depth profiles of Chl a-specific uptake rates for ammonium and nitrate paralleled those of absolute uptake rates, i.e. values of VNH 4+Chl were highest (up to 16.1 nmol μg -1 h -1) in nitrate-poor surface waters while values of VNO 3-Chl were maximum (up to 8.4 nmol μg -1 h -1)within the nitracline. This latter vertical ordering of planktonic nitrogen nutrition was consistent with an aged upwelling situation. However, applying several indices of cell metabolism and nutritional status, such as 15N uptake/enzyme activity, surge uptake internally controlled uptake, and V maxChl/K t ratios, we were able to demonstrate that the phytoplankton assemblages inhabiting the nutrient-impoverished upper layer still bore the signature of physically mediated nitrogen (nitrate) supply generated by active upwelling that had occurred during the week before our visit to the area. This signature was the most evident in samples from the station furthest inshore and faded with distance from shore as a result of the deepening of the nitrate isopleths (weakening of upwelling activity), which showed the same offshore trend. The appearance of nitrate-rich waters at the surface, after a strong pulse of upwelling favourable winds just before the end of the cruise, led to a five-fold increase in average (over the euphotic zone

  12. Isolation of coniferyl esters from Capsicum baccatum L., and their enzymatic preparation and agonist activity for TRPV1.

    Science.gov (United States)

    Kobata, Kenji; Tate, Hitomi; Iwasaki, Yusaku; Tanaka, Yoshiyuki; Ohtsu, Keigo; Yazawa, Susumu; Watanabe, Tatsuo

    2008-03-01

    Coniferyl esters--capsiconiate and dihydrocapsiconiate--were isolated from the fruits of the pepper, Capsicum baccatum L. var. praetermissum. Their structures were determined by spectroscopic methods to be coniferyl (E)-8-methyl-6-nonenoate (capsiconiate) and coniferyl 8-methylnonanoate (dihydrocapsiconiate). This finding was further confirmed by the lipase-catalyzed condensation of coniferyl alcohol with its corresponding fatty acid derivative. The agonist activity of the esters for transient receptor potential vanilloid 1 (TRPV1) was evaluated by conducting an analysis of the intracellular calcium concentrations in TRPV1-expressing HEK293 cells. The EC50 values of capsiconiate and dihydrocapsiconiate were 3.2 and 4.2 microM, respectively. PMID:18190936

  13. NMR Analysis of a Novel Enzymatically Active Unlinked Dengue NS2B-NS3 Protease Complex*

    Science.gov (United States)

    Kim, Young Mee; Gayen, Shovanlal; Kang, CongBao; Joy, Joma; Huang, Qiwei; Chen, Angela Shuyi; Wee, John Liang Kuan; Ang, Melgious Jin Yan; Lim, Huichang Annie; Hung, Alvin W.; Li, Rong; Noble, Christian G.; Lee, Le Tian; Yip, Andy; Wang, Qing-Yin; Chia, Cheng San Brian; Hill, Jeffrey; Shi, Pei-Yong; Keller, Thomas H.

    2013-01-01

    The dengue virus (DENV) is a mosquito-borne pathogen responsible for an estimated 100 million human infections annually. The viral genome encodes a two-component trypsin-like protease that contains the cofactor region from the nonstructural protein NS2B and the protease domain from NS3 (NS3pro). The NS2B-NS3pro complex plays a crucial role in viral maturation and has been identified as a potential drug target. Using a DENV protease construct containing NS2B covalently linked to NS3pro via a Gly4-Ser-Gly4 linker (“linked protease”), previous x-ray crystal structures show that the C-terminal fragment of NS2B is remote from NS3pro and exists in an open state in the absence of an inhibitor; however, in the presence of an inhibitor, NS2B complexes with NS3pro to form a closed state. This linked enzyme produced NMR spectra with severe signal overlap and line broadening. To obtain a protease construct with a resolved NMR spectrum, we expressed and purified an unlinked protease complex containing a 50-residue segment of the NS2B cofactor region and NS3pro without the glycine linker using a coexpression system. This unlinked protease complex was catalytically active at neutral pH in the absence of glycerol and produced dispersed cross-peaks in a 1H-15N heteronuclear single quantum correlation spectrum that enabled us to conduct backbone assignments using conventional techniques. In addition, titration with an active-site peptide aldehyde inhibitor and paramagnetic relaxation enhancement studies demonstrated that the unlinked DENV protease exists predominantly in a closed conformation in solution. This protease complex can serve as a useful tool for drug discovery against DENV. PMID:23511634

  14. Effect of permethrin, anthracene and mixture exposure on shell components, enzymatic activities and proteins status in the Mediterranean clam Venerupis decussata

    International Nuclear Information System (INIS)

    Highlights: • We assessed toxicity of anthracene, permethrin and their mixture on clams. • Tissue and stressor-dependent changes were observed in biochemical responses. • Permethrin induces phase transition from aragonite to calcite in shell structure. • Interactive effects were observed on digestive gland and gill biomarkers. • Both approaches give new vision to risk assessment of organic pollution. - Abstract: Anthracene (ANT) and permethrin (PER) are two of the more toxic compounds reaching the marine environment. This study aimed to determine the impact of these molecules on Venerupis decussata, an economically important species cultured on the Tunisian coast. Shell structure and its possible transformation upon exposure to the two contaminants were studied by X-ray diffraction and gravimetric analyses. Results revealed a phase transition in shell composition from aragonite to calcite after PER exposure, to a mixture of PER and ANT (Mix) but not for ANT alone. Catalase (CAT), superoxide dismutase (SOD) and glutathione transferase (GST) activities were determined in digestive gland and gills after exposure to ANT, PER and Mix to assess the impact of the contamination on the oxidative status of V. decussata. Enzyme activities increased in the digestive gland after PER treatment and in the gills after ANT treatment. PER exposure significantly reduced the levels of free thiols and increased levels of carbonylated proteins in the digestive gland, as compared to controls. In contrast, ANT exposure significantly reduced free thiols and increased the number of carbonylated proteins in the gills. Mix induced additive effects as measured by both enzymatic and proteomic approaches. The present study suggests that PER has a strong effect on shell structure; that PER and ANT exposure generate compound-dependent oxidative stress in the tissues of V. decussata and that a mixture of the two compounds has synergistic effects on biochemical response

  15. Effect of permethrin, anthracene and mixture exposure on shell components, enzymatic activities and proteins status in the Mediterranean clam Venerupis decussata

    Energy Technology Data Exchange (ETDEWEB)

    Sellami, Badreddine, E-mail: sellamibadreddine@gmail.com [Laboratory of Environment Biomonitoring, Coastal Ecology Unit, Faculty of Sciences of Bizerta, University of Carthage, 7021 Zarzouna (Tunisia); Khazri, Abdelhafidh [Laboratory of Environment Biomonitoring, Coastal Ecology Unit, Faculty of Sciences of Bizerta, University of Carthage, 7021 Zarzouna (Tunisia); Mezni, Amine [Unit of Research 99/UR12-30, Department of Chemistry, Faculty of Sciences of Bizerte, 7021 Jarzouna (Tunisia); Louati, Héla; Dellali, Mohamed; Aissa, Patricia; Mahmoudi, Ezzeddine; Beyrem, Hamouda [Laboratory of Environment Biomonitoring, Coastal Ecology Unit, Faculty of Sciences of Bizerta, University of Carthage, 7021 Zarzouna (Tunisia); Sheehan, David, E-mail: d.sheehan@ucc.ie [Environmental Research Institute and Department of Biochemistry, University College Cork, Western Gateway Building, Western Road, Cork (Ireland)

    2015-01-15

    Highlights: • We assessed toxicity of anthracene, permethrin and their mixture on clams. • Tissue and stressor-dependent changes were observed in biochemical responses. • Permethrin induces phase transition from aragonite to calcite in shell structure. • Interactive effects were observed on digestive gland and gill biomarkers. • Both approaches give new vision to risk assessment of organic pollution. - Abstract: Anthracene (ANT) and permethrin (PER) are two of the more toxic compounds reaching the marine environment. This study aimed to determine the impact of these molecules on Venerupis decussata, an economically important species cultured on the Tunisian coast. Shell structure and its possible transformation upon exposure to the two contaminants were studied by X-ray diffraction and gravimetric analyses. Results revealed a phase transition in shell composition from aragonite to calcite after PER exposure, to a mixture of PER and ANT (Mix) but not for ANT alone. Catalase (CAT), superoxide dismutase (SOD) and glutathione transferase (GST) activities were determined in digestive gland and gills after exposure to ANT, PER and Mix to assess the impact of the contamination on the oxidative status of V. decussata. Enzyme activities increased in the digestive gland after PER treatment and in the gills after ANT treatment. PER exposure significantly reduced the levels of free thiols and increased levels of carbonylated proteins in the digestive gland, as compared to controls. In contrast, ANT exposure significantly reduced free thiols and increased the number of carbonylated proteins in the gills. Mix induced additive effects as measured by both enzymatic and proteomic approaches. The present study suggests that PER has a strong effect on shell structure; that PER and ANT exposure generate compound-dependent oxidative stress in the tissues of V. decussata and that a mixture of the two compounds has synergistic effects on biochemical response.

  16. Genome-Wide Identification, 3D Modeling, Expression and Enzymatic Activity Analysis of Cell Wall Invertase Gene Family from Cassava (Manihot esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Yuan Yao

    2014-04-01

    Full Text Available The cell wall invertases play a crucial role on the sucrose metabolism in plant source and sink organs. In this research, six cell wall invertase genes (MeCWINV1-6 were cloned from cassava. All the MeCWINVs contain a putative signal peptide with a predicted extracellular location. The overall predicted structures of the MeCWINV1-6 are similar to AtcwINV1. Their N-terminus domain forms a β-propeller module and three conserved sequence domains (NDPNG, RDP and WECP(VD, in which the catalytic residues are situated in these domains; while the C-terminus domain consists of a β-sandwich module. The predicted structure of Pro residue from the WECPD (MeCWINV1, 2, 5, and 6, and Val residue from the WECVD (MeCWINV3 and 4 are different. The activity of MeCWINV1 and 3 were higher than other MeCWINVs in leaves and tubers, which suggested that sucrose was mainly catalyzed by the MeCWINV1 and 3 in the apoplastic space of cassava source and sink organs. The transcriptional levels of all the MeCWINVs and their enzymatic activity were lower in tubers than in leaves at all the stages during the cassava tuber development. It suggested that the major role of the MeCWINVs was on the regulation of carbon exportation from source leaves, and the ratio of sucrose to hexose in the apoplasts; the role of these enzymes on the sucrose unloading to tuber was weaker.

  17. Opioid Antagonist Activities of Enzymatic Hydrolysates from Rice Bran Protein%米糠蛋白酶解物类阿片拮抗活性的研究

    Institute of Scientific and Technical Information of China (English)

    陈季旺; 孙庆杰; 姚惠源; 夏文水

    2005-01-01

    采用水解度(DH)对酶解米糠蛋白进行了研究,比较六种蛋白酶对米糠可溶性蛋白的水解作用,结果发现胰蛋白酶对米糠可溶性蛋白的水解作用最强,它的最适作用条件为:pH8.0,温度为37℃,[E]/[S]为12.5usp-u/kg.采用离体豚鼠回肠(GPI)检定法测定上述酶解物的类阿片拮抗活性,结果发现胰蛋白酶水解产物具有明显的类阿片拮抗活性,DH为11.9%时,其水解产物(样品A)的类阿片拮抗活性最高.体积排阻高效液相色谱测定它的相对分子质量分布范围在125~24233Da之间.%The enzymatic hydrolysis of rice bran protein was studied with degree of hydrolysis. Trypsin was found to be the most effective protease and its optimum conditions were pH8, ratio of enzyme to substrate 1:100 and temperature of 37℃.Resulting hydrolysates were tested for their opioid antagonist activities based on guinea pig ileum-contracting assayss. The hydrolysate produced at 11.9% of degree of hydrolysis showed the highest opioid antagonist activity. High performance size exclusion chromatography results indicated that its molecular weight distribution mainly ranged from 125~24233Da.

  18. Partial root zone drying: regulation of photosynthetic limitations and antioxidant enzymatic activities in young olive (Olea europaea) saplings.

    Science.gov (United States)

    Aganchich, Badia; Wahbi, Said; Loreto, Francesco; Centritto, Mauro

    2009-05-01

    The effect of partial root drying (PRD) irrigation on split-root olive (Olea europaea L. cv Picholine marocaine) saplings was investigated. An irrigated control and two PRD regimes were applied (control: irrigation applied on both sides of the root system to keep the soil water content close to field capacity; PRD(50): irrigation applied at 50% of the control amount on one side of the root system and irrigation withheld from the other side, with irrigation regimes switched between the sides of the root system every 2 weeks; and PRD(100): irrigation applied at 100% of the control amount on one side and irrigation withheld on the other side, with irrigation regimes switched between the sides of the root system every 2 weeks. Only saplings in the PRD(50) regime were subjected to water-deficit irrigation. The PRD treatments significantly affected water relations and vegetative growth throughout the growing season. Predawn leaf water potential and relative water content differed significantly between the PRD(50) and PRD(100) saplings, leading to reduced stomatal conductance, carbon assimilation, shoot length and leaf number in PRD(50) saplings. However, the PRD(50) water-deficit treatment did not affect the capacity of the saplings to assimilate CO(2). Activities of superoxide dismutase, soluble and insoluble peroxidase (POX) and polyphenol oxidase were up-regulated by the PRD(50) and PRD(100) treatments compared with control values. The higher activities of both soluble and insoluble POX observed in PRD(50) saplings may reflect the greater inhibitory effect of this treatment on vegetative growth. Up-regulation of the detoxifying systems in the PRD(100) and PRD(50) saplings may have provided protection mechanisms against irreversible damage to the photosynthetic machinery, thereby allowing the photosynthetic apparatus to function and preventing the development of severe water stress. We also measured CO(2) assimilation rate/internal leaf CO(2) concentration (A

  19. Overexpression of Scg5 increases enzymatic activity of PCSK2 and is inversely correlated with body weight in congenic mice

    Directory of Open Access Journals (Sweden)

    Islas-Trejo Alma

    2008-04-01

    Full Text Available Abstract Background The identification of novel genes is critical to understanding the molecular basis of body weight. Towards this goal, we have identified secretogranin V (Scg5; also referred to as Sgne1, as a candidate gene for growth traits. Results Through a combination of DNA microarray analysis and quantitative PCR we identified a strong expression quantitative trait locus (eQTL regulating Scg5 expression in two mouse chromosome 2 congenic strains and three additional F2 intercrosses. More importantly, the eQTL was coincident with a body weight QTL in congenic mice and Scg5 expression was negatively correlated with body weight in two of the F2 intercrosses. Analysis of haplotype blocks and genomic sequencing of Scg5 in high (C3H/HeJ, DBA/2J, BALB/cByJ, CAST/EiJ and low (C57BL/6J expressing strains revealed mutations unique to C57BL/6J and possibly responsible for the difference in mRNA abundance. To evaluate the functional consequence of Scg5 overexpression we measured the pituitary levels of 7B2 protein and PCSK2 activity and found both to be increased. In spite of this increase, the level of pituitary α-MSH, a PCSK2 processing product, was unaltered. Conclusion Together, these data support a role for Scg5 in the modulation of body weight.

  20. Cloning and expression of the enzymatic region of Streptococcal hyaluronidase

    OpenAIRE

    Nafiseh Al-Sadat Mirjamali; Safieh Soufian; Neda Molaee; Shabnam Sadoogh Abbasian; Hamid Abtahi

    2014-01-01

    Objective(s): Streptococcus pyogenes produces extracellular hyaluronidase enzyme. This enzyme is directly associated with the spread of the organism during infection. The objective of the present study was to clone and express the nucleotide sequence of the enzyme which is involved in hyaluronidase enzymatic activity. Materials and Methods: The enzymatic region of hyaluronidase gene was detected by bioinformatics method. The PCR method was used to amplify enzymatic region of hyaluronidase gen...

  1. Parp1 localizes within the Dnmt1 promoter and protects its unmethylated state by its enzymatic activity.

    Directory of Open Access Journals (Sweden)

    Michele Zampieri

    Full Text Available BACKGROUND: Aberrant hypermethylation of CpG islands in housekeeping gene promoters and widespread genome hypomethylation are typical events occurring in cancer cells. The molecular mechanisms behind these cancer-related changes in DNA methylation patterns are not well understood. Two questions are particularly important: (i how are CpG islands protected from methylation in normal cells, and how is this protection compromised in cancer cells, and (ii how does the genome-wide demethylation in cancer cells occur. The latter question is especially intriguing since so far no DNA demethylase enzyme has been found. METHODOLOGY/PRINCIPAL FINDINGS: Our data show that the absence of ADP-ribose polymers (PARs, caused by ectopic over-expression of poly(ADP-ribose glycohydrolase (PARG in L929 mouse fibroblast cells leads to aberrant methylation of the CpG island in the promoter of the Dnmt1 gene, which in turn shuts down its transcription. The transcriptional silencing of Dnmt1 may be responsible for the widespread passive hypomethylation of genomic DNA which we detect on the example of pericentromeric repeat sequences. Chromatin immunoprecipitation results show that in normal cells the Dnmt1 promoter is occupied by poly(ADP-ribosylated Parp1, suggesting that PARylated Parp1 plays a role in protecting the promoter from methylation. CONCLUSIONS/SIGNIFICANCE: In conclusion, the genome methylation pattern following PARG over-expression mirrors the pattern characteristic of cancer cells, supporting our idea that the right balance between Parp/Parg activities maintains the DNA methylation patterns in normal cells. The finding that in normal cells Parp1 and ADP-ribose polymers localize on the Dnmt1 promoter raises the possibility that PARylated Parp1 marks those sequences in the genome that must remain unmethylated and protects them from methylation, thus playing a role in the epigenetic regulation of gene expression.

  2. ATIVIDADE ENZIMÁTICA DE MICRORGANISMOS ISOLADOS DO JACATUPÉ (Pachyrhizus erosus L. Urban ENZYMATIC ACTIVITY OF MICROORGANISMS ISOLATED FROM YAM BEAN LEGUME (Pachyrhizus erosus L. Urban

    Directory of Open Access Journals (Sweden)

    Tânia L. Montenegro STAMFORD

    1998-10-01

    Full Text Available O isolamento e a identificação de microrganismos produtores de enzimas de interesse comercial, utilizando tubérculos de jacatupé (Pachyrhizus erosus L. Urban, foi o objetivo principal deste trabalho. Isolaram-se microrganismos endofíticos e epifíticos identificados por observação micromorfológica. A avaliação da atividade enzimática das linhagens foi determinada pelo método de difusão em ágar. As sessenta e oito linhagens isoladas dos tubérculos de jacatupé foram cultivadas em meio sólido específico para amilase, lipase, protease e celulase por 96h a 280 C. Os microrganismos epifíticos encontrados foram Pithomyces (7,3%, Aspergillus (19,2%, Fusarium (5,9% e Trichoderma (5,8%, e os endofíticos foram Mucor (7,3%, Rhizopus (10,3%, Bacillus (19,0%, Staphylococcus (10,3% e Nocardiopsis (15%. As linhagens de Nocardiopsis sp. apresentaram atividade lipolítica superior à do padrão, porém a atividade amilolítica não apresentou diferença significativa comparada com o padrão. As linhagens de Mucor sp., Pithomyces sp. e Staphylococcus sp. produziram atividade proteolítica abaixo do padrão. Nenhum isolado apresentou atividade celulolítica.The isolation and identification of microorganisms that produce enzyme of commercial interest utilizing tubers of yam bean legume (Pachyrrizus erosus L. Urban was the main objective of this work. Endophytic and epiphytic microorganisms were isolated by micromorphologyc observation. The agar diffusion method was used to determine the enzymatic activity. Sixty-eight isolates from yam bean tubers were cultured at 280 C in solid medium specific to amylase, lipase, protease and cellulase for 96h. The epiphytic microorganisms Pithomyces (7,3%, Aspergillus (19,2%, Fusarium (5,9% and Trichoderma (5,8% and the endophytic microorganisms Mucor (7,3%, Rhizopus (10,3% Bacillus (19%, Staphylococcus (10,3% and Nocardiopsis (15% were isolated. Compared to the specific standard culture Nocardiopsis sp. showed

  3. Antioxidant Enzymatic Activities and Gene Expression Associated with Heat Tolerance in the Stems and Roots of Two Cucurbit Species (“Cucurbita maxima” and “Cucurbita moschata”) and Their Interspecific Inbred Line “Maxchata”

    OpenAIRE

    Neelam Ara; Korakot Nakkanong; Wenhui Lv; Jinghua Yang; Zhongyuan Hu; Mingfang Zhang

    2013-01-01

    The elucidation of heat tolerance mechanisms is required to combat the challenges of global warming. This study aimed to determine the antioxidant enzyme responses to heat stress, at the enzymatic activity and gene expression levels, and to investigate the antioxidative alterations associated with heat tolerance in the stems and roots of squashes using three genotypes differing in heat tolerance. Plants of heat-tolerant “C. moschata”, thermolabile “C. maxima” and moderately heat-tolerant inte...

  4. Comparative evaluation of tableting compression behaviors by methods of internal and external lubricant addition: Inhibition of enzymatic activity of trypsin preparation by using external lubricant addition during the tableting compression process

    OpenAIRE

    Otsuka, Makoto; Sato, Mitsuyo; Matsuda, Yoshihisa

    2001-01-01

    This study evaluated tableting compression by using internal and external lubricant addition. The effect of lubricant addition on the enzymatic activity of trypsin, which was used as a model drug during the tableting compression process, was also investigated. The powder mixture (2% crystalline trypsin, 58% crystalline lactose, and 40% microcrystalline cellulose) was kneaded with 5% hydroxypropyl cellulose aqueous solution and then granulated using an extruding granulator equipped with a 0.5-...

  5. Enzymatic activity of lipase in post-metamorphic phase bullfrogs Atividade enzimática da lipase em rã-touro na fase pós-metamórfica

    OpenAIRE

    Luís Gustavo Tavares Braga; Maria Goreti de Almeida Oliveira; William Cardoso Lima; Ricardo Frederico Euclydes

    2006-01-01

    The knowledge of the digestive system of bullfrogs is an important step for the determination of their nutritional requirements throughout growth phases. With the objective of evaluating the enzymatic activity of lipase in the intestinal content of bullfrogs (Rana catesbeiana Shaw, 1802), 100 animals with median weight of 3.6 g were distributed in stalls under controlled temperature and photoperiod. The frogs, selected at the post-metamorphic phase, received commercial extruded diet ad libitu...

  6. Effect of aflatoxin B1 on growth and enzymatic activity of a native strain of Bacillus sp Efecto de la aflatoxina B1 sobre el crecimiento y actividad proteolítica de una cepa nativa de Bacillus sp

    Directory of Open Access Journals (Sweden)

    Márquez Edna Judith

    2004-07-01

    Full Text Available The effect of different aflatoxin B1 (AFAB1 concentrations on alkaline protease growth and enzymatic activity was evaluated; a native strain of alkalophilic Bacillus sp cultivated in CSL (Corn Steep Liquor was used. It was found that the effect of AFAB1 on the strain inhibited its growth and enzymatic activity to 1 ppm, showing that the strain is highly sensible to AFAB1, meaning that medium obtained f rom Colombian corn contaminated with this mycotoxin cannot be easily used. Concentrations less than 0.1 ppm did not affect growth and enzymatic activity. Key words: Bacillus, aflatoxin, alkaline proteases.Se evaluó el efecto de diferentes concentraciones de aflatoxina B1 (AFAB1 sobre el crecimiento y actividad enzimática de proteasas alcalinas de una cepa nativa de Bacillus sp Alcalofílico cultivada en LAM (Licor Agotado de Maíz. Se encontró que la cepa inhibe su crecimiento y actividad enzimática a 1 ppm, lo que demuestra una alta sensibilidad de la cepa evaluada a la AFAB1 e imposibilita utilizar fácilmente medios obtenidos de maíz nacional contaminado con esta micotoxina. Las concentraciones inferiores a 0.1 ppm no tienen ningún efecto sobre el crecimiento y la actividad enzimática. Palabras clave: Bacillus, aflatoxina, proteasas alcalinas.

  7. Studies on gene structure, enzymatic activity and regulatory mechanism of acetohydroxy acid isomeroreductase from G2 pea

    Institute of Scientific and Technical Information of China (English)

    XU; Yunjian; (

    2003-01-01

    , P., Smith, J. K. et al., Regulation of tobacco acetolactate synthase gene expression, Plant Physiol., 1993, 102: 1009-1018.[21]Singh, B. K., Newhouse, K. E., Stidham, M. A. et al., Actohydroxyacid synthase-imidazolinone interaction, in Biosynthesis of Branched Chain Amino Acids (eds. Barak, Z., Chipman, D. M., Schloss, J. V.), Weinheim: VCH, 1990, 357-372.[22]Stidham, M. A., Singh, B. K., Imidazolinone-acetohydroxyacid synthase interactions, in The Imidazolinone Herbicides (eds. Shaner, L. L., O'Connor, S. L.), Boca Raton, F L: CRC Press, 1991, 71-90.[23]Szamosi, I. T., Shaner, D. L., Singh, B. K., Identification and characterization of a biodegradative form of threonine deghdratase in senescing tomato leaf, Plant Physiol., 1993, 101: 999-1004.[24]Hofgen, R., Laber, B., Schuttke, I. et al., Repression of acetolactate synthase activity through antisense inhibition: Molecular and biochemical analysis of transgenic potato (Solanum tuberosum L. cv Desiree) plants, Plant Physiol., 1995, 107: 469-477.

  8. Enzymatic hydrolysis of potato pulp

    Directory of Open Access Journals (Sweden)

    Mariusz Lesiecki

    2012-03-01

    Full Text Available Background. Potato pulp constitutes a complicated system of four types of polysaccharides: cellulose, hemicellulose, pectin and starch. Its composition makes it a potential and attractive raw material for the production of the second generation bioethanol. The aim of this research project was to assess the usefulness of commercial enzymatic preparations for the hydrolysis of potato pulp and to evaluate the effectiveness of hydrolysates obtained in this way as raw materials for ethanol fermentation. Material  and methods. Sterilised potato pulp was subjected to hydrolysis with commercial enzymatic preparations. The effectiveness of the preparations declared as active towards only one fraction of potato pulp (separate amylase, pectinase and cellulase activity and mixtures of these preparations was analysed. The monomers content in hydrolysates was determined using HPLC method. Results.  The application of amylolytic enzymes for potato pulp hydrolysis resulted in the release of only 18% of raw material with glucose as the dominant (77% constituent of the formed product. In addition, 16% galactose was also determined in it. The hydrolysis of the cellulose fraction yielded up to 35% raw material and the main constituents of the obtained hydrolysate were glucose (46% and arabinose (40%. Simultaneous application of amylolytic, cellulolytic and pectinolytic enzymes turned out to be the most effective way of carrying out the process as its efficiency in this case reached 90%. The obtained hydrolysate contained 63% glucose, 25% arabinose and 12% other simple substances. Conclusion. The application of commercial enzymatic preparations made it possible to perform potato pulp hydrolysis with 90% effectiveness. This was achieved by the application of a complex of amylolytic, cellulolytic and pectinolytic enzymes and the hydrolysate obtained in this way contained, primarily, glucose making it a viable substrate for ethanol fermentation.

  9. Enzymatic synthesis of vanillin

    NARCIS (Netherlands)

    van den Heuvel, RHH; Fraaije, MW; Laane, C; van Berkel, WJH; Heuvel, Robert H.H. van den; Berkel, Willem J.H. van

    2001-01-01

    Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated conv

  10. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    -efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin have been studied. Sphingomyelin (SM) is a ubiquitous membrane-lipid and dairy products or by-products is a rich source of sphingomyelin. In...

  11. Study on the effect of prostaglandin F2α treatment on semen characteristics and enzymatic activates of Awassi rams in breeding and non breeding seasons

    Directory of Open Access Journals (Sweden)

    Osama Ibrahim Azawi,

    2011-05-01

    non breeding season (52.34 ± 8.96 and 57.43 ± 19.9 vs. 117.02 ± 5.26 and 131.88 ± 5.01, respectively. Other enzymatic activities including ALT, AST, ACP and ALP showed no significant differences between Awassi rams treated with PGF2α and control groups in both breeding and non breeding seasons. In conclusion, PGF2αtreatment of Awassi rams improved sperm concentration and testicular volume

  12. Non-enzymatic modifications of prostaglandin H synthase 1 affect bifunctional enzyme activity - Implications for the sensitivity of blood platelets to acetylsalicylic acid.

    Science.gov (United States)

    Kassassir, Hassan; Siewiera, Karolina; Talar, Marcin; Stec-Martyna, Emilia; Pawlowska, Zofia; Watala, Cezary

    2016-06-25

    Due to its ability to inhibit the blood platelet PGHS-1, acetylsalicylic acid (ASA, Aspirin(®)) is widely used as a preventive agent in atherothrombotic diseases. However, its beneficial effects seem to be lower in diabetic patients, suggesting that protein glycation may impair effective ASA-mediated acetylation process. On the other hand, it is proposed that ASA can prevent some of the late complications of diabetes by lowering the extent of glycation at protein free amino groups. The aim of this work was to evaluate the extents of non-enzymatic N-glycosylation (glycation) and acetylation of blood platelet PGHS-1 (COX-1) and the competition between glycation and acetylation was investigated in order to demonstrate how these two reactions may compete against platelet PGHS-1. When PGHS-1 was incubated with glycating/acetylating agents (glucose, Glu; 1,6-bisphosphofructose, 1,6-BPF; methylglyoxal, MGO, acetylsalicylic acid, ASA), the enzyme was modified in 13.4 ± 1.6, 5.3 ± 0.5, 10.7 ± 1.2 and 6.4 ± 1.1 mol/mol protein, respectively, and its activity was significantly reduced. The prior glycation/carbonylation of PGHS-1 with Glu, 1,6-BPF or MGO decreased the extent of acetylation from 6.4 ± 1.1 down to 2.5 ± 0.2, 3.6 ± 0.3 and 5.2 ± 0.2 mol/mol protein, respectively, but the enzyme still remained susceptible to the subsequent inhibition of its activity with ASA. When PGHS-1 was first acetylated with ASA and then incubated with glycating/carbonylating agents, we observed the following reductions in the enzyme modifications: from 13.4 ± 1.6 to 8.7 ± 0.6 mol/mol protein for Glu, from 5.3 ± 0.5 to 3.9 ± 0.3 mol/mol protein for 1,6-BPF and from 10.7 ± 1.2 to 7.5 ± 0.5 mol/mol protein for MGO, however subsequent glycation/carbonylation did not significantly affect PGHS-1 function. Overall, our outcomes allow to better understand the structural aspects of the chemical competition between glycation and acetylation of PGHS-1

  13. Integrated reactor concepts for the enzymatic kinetic synthesis of cephalexin

    NARCIS (Netherlands)

    Schroën, C.G.P.H.; Nierstrasz, V.A.; Bosma, R.; Kroon, P.J.; Tjeerdsma, P.S.; DeVroom, E.; VanderLaan, J.M.; Moody, H.M.; Beeftink, H.H.; Janssen, A.E.M.; Tramper, J.

    2002-01-01

    Integrated process concepts for enzymatic cephalexin synthesis were investigated by our group, and this article focuses on the integration of reactions and product removal during the reactions. The last step in cephalexin production is the enzymatic kinetic coupling of activated phenylglycine (pheny

  14. Biofunctional Properties of Enzymatic Squid Meat Hydrolysate

    OpenAIRE

    Choi, Joon Hyuk; Kim, Kyung-Tae; Kim, Sang Moo

    2015-01-01

    Squid is one of the most important commercial fishes in the world and is mainly utilized or consumed as sliced raw fish or as processed products. The biofunctional activities of enzymatic squid meat hydrolysate were determined to develop value-added products. Enzymatic squid hydrolysate manufactured by Alcalase effectively quenched 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and hydrogen peroxide radical with IC50 values of 311, 3,410, and 111.5 μg/mL, respectively. Angiotensin I...

  15. Enzymatic treatment of estrogens and estrogen glucuronide

    Institute of Scientific and Technical Information of China (English)

    Takaaki Tanaka; Toshiyuki Tamura; Yuuichi Ishizaki; Akito Kawasaki; Tomokazu Kawase; Masahiro Teraguchi; Masayuki Taniguchi

    2009-01-01

    Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers.In this article we investigated the enzymatic treatment of three estrogens (estrone,17β-estradiol,and 17α-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen.The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay.In addition,we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-(β-D-glucuronide) degradation.The two enzymes eliminated 17β-estradiol 3-(β-D-glucuronide) and the intermediate,17β-estradiol,efficiently.

  16. Enzymatic desulfurization of coal

    Energy Technology Data Exchange (ETDEWEB)

    Boyer, Y.N.; Crooker, S.C.; Kitchell, J.P.; Nochur, S.V.

    1991-05-16

    The overall objective of this program was to investigate the feasibility of an enzymatic desulfurization process specifically intended for organic sulfur removal from coal. Toward that end, a series of specific objectives were defined: (1) establish the feasibility of (bio)oxidative pretreatment followed by biochemical sulfate cleavage for representative sulfur-containing model compounds and coals using commercially-available enzymes; (2) investigate the potential for the isolation and selective use of enzyme preparations from coal-utilizing microbial systems for desulfurization of sulfur-containing model compounds and coals; and (3) develop a conceptual design and economic analysis of a process for enzymatic removal of organic sulfur from coal. Within the scope of this program, it was proposed to carry out a portion of each of these efforts concurrently. (VC)

  17. Coated tube for immunochemical and enzymatic assays

    International Nuclear Information System (INIS)

    Containers such as test tubes suitable for use in solid phase immunochemical, enzymatical and particularly radioimmunoassay procedures are described. The lower part of the tube is a polymer, coated with an inert protein to which a biologically active substance eg an antibody to triiodothyronine, thyroxine or digoxin, is attached. (U.K.)

  18. Induction of indoleamine 2,3-dioxygenase (IDO) enzymatic activity contributes to interferon-gamma induced apoptosis and death receptor 5 expression in human non-small cell lung cancer cells.

    Science.gov (United States)

    Chung, Ting Wen; Tan, Kok-Tong; Chan, Hong-Lin; Lai, Ming-Derg; Yen, Meng-Chi; Li, Yi-Ron; Lin, Sheng Hao; Lin, Chi-Chen

    2014-01-01

    Interferon-gamma (IFN-γ) has been used to treat various malignant tumors. However, the molecular mechanisms underlying the direct anti-proliferative activity of IFN-γ are poorly understood. In the present study, we examined the in vitro antitumor activity of IFN-γ on two human non-small-cell lung carcinoma (NSCLC) cell lines, H322M and H226. Our findings indicated that IFN-γ treatment caused a time-dependent reduction in cell viability and induced apoptosis through a FADD-mediated caspase-8/tBid/mitochondria-dependent pathway in both cell lines. Notably, we also postulated that IFN-γ increased indoleamine 2,3-dioxygenase (IDO) expression and enzymatic activity in H322M and H226 cells. In addition, inhibition of IDO activity by the IDO inhibitor 1-MT or tryptophan significantly reduced IFN-γ-induced apoptosis and death receptor 5 (DR5) expression, which suggests that IDO enzymatic activity plays an important role in the anti-NSCLC cancer effect of IFN-γ. These results provide new mechanistic insights into interferon-γ antitumor activity and further support IFN-γ as a potential therapeutic adjuvant for the treatment of NCSLC. PMID:25292102

  19. Enzymatic synthesis of vanillin

    OpenAIRE

    van den Heuvel, RHH; Fraaije, MW; Laane, C; van Berkel, WJH; Heuvel, Robert H.H. van den; van Berkel, Willem J. H.

    2001-01-01

    Due to increasing interest in natural vanillin, two enzymatic routes for the synthesis of vanillin were developed. The flavoprotein vanillyl alcohol oxidase (VAO) acts on a wide range of phenolic compounds and converts both creosol and vanillylamine to vanillin with high yield. The VAO-mediated conversion of creosol proceeds via a two-step process in which the initially formed vanillyl alcohol is further oxidized to vanillin. Catalysis is limited by the formation of an abortive complex betwee...

  20. Unravelling the importance of forest age stand and forest structure driving microbiological soil properties, enzymatic activities and soil nutrients content in Mediterranean Spanish black pine(Pinus nigra Ar. ssp. salzmannii) Forest.

    Science.gov (United States)

    Lucas-Borja, M E; Hedo, J; Cerdá, A; Candel-Pérez, D; Viñegla, B

    2016-08-15

    This study aimed to investigate the effects that stand age and forest structure have on microbiological soil properties, enzymatic activities and nutrient content. Thirty forest compartments were randomly selected at the Palancares y Agregados managed forest area (Spain), supporting forest stands of five ages; from 100 to 80years old to compartments with trees that were 19-1years old. Forest area ranging from 80 to 120years old and without forest intervention was selected as the control. We measured different soil enzymatic activities, soil respiration and nutrient content (P, K, Na, Mg, Cr, Mn, Fe, Co, Ni, Cu, Zn, Pb and Ca) in the top cm of 10 mineral soils in each compartment. Results showed that the lowest forest stand age and the forest structure created by management presented lower values of organic matter, soil moisture, water holding capacity and litterfall and higher values of C/N ratio in comparison with the highest forest stand age and the related forest structure, which generated differences in soil respiration and soil enzyme activities. The forest structure created by no forest management (control plot) presented the highest enzymatic activities, soil respiration, NH4(+) and NO3(-). Results did not show a clear trend in nutrient content comparing all the experimental areas. Finally, the multivariate PCA analysis clearly clustered three differentiated groups: Control plot; from 100 to 40years old and from 39 to 1year old. Our results suggest that the control plot has better soil quality and that extreme forest stand ages (100-80 and 19-1years old) and the associated forest structure generates differences in soil parameters but not in soil nutrient content. PMID:27099995

  1. Apoptosis induction in human breast cancer (MCF-7) cells by a novel venom L-amino acid oxidase (Rusvinoxidase) is independent of its enzymatic activity and is accompanied by caspase-7 activation and reactive oxygen species production.

    Science.gov (United States)

    Mukherjee, Ashis K; Saviola, Anthony J; Burns, Patrick D; Mackessy, Stephen P

    2015-10-01

    We report the elucidation of a mechanism of apoptosis induction in breast cancer (MCF-7) cells by an L-amino acid oxidase (LAAO), Rusvinoxidase, purified from the venom of Daboia russelii russelii. Peptide mass fingerprinting analysis of Rusvinoxidase, an acidic monomeric glycoprotein with a mass of ~57 kDa, confirmed its identity as snake venom LAAO. The enzymatic activity of Rusvinoxidase was completely abolished after two cycles of freezing and thawing; however, its cytotoxicity toward MCF-7 cells remained unaffected. Dose- and time-dependent induction of apoptosis by Rusvinoxidase on MCF-7 cells was evident from changes in cell morphology, cell membrane integrity, shrinkage of cells and apoptotic body formation accompanied by DNA fragmentation. Rusvinoxidase induced apoptosis in MCF-7 cells by both the extrinsic (death-receptor) and intrinsic (mitochondrial) signaling pathways. The former pathway of apoptosis operated through activation of caspase-8 that subsequently activated caspase-7 but not caspase-3. Rusvinoxidase-induced intrinsic pathway of apoptosis was accompanied by a time-dependent depolarization of the mitochondrial membrane through the generation of reactive oxygen species, followed by a decrease in cellular glutathione content and catalase activity, and down-regulation of expression of anti-apoptotic proteins Bcl-XL and heat-shock proteins (HSP-90 and HSP-70). Rusvinoxidase treatment resulted in increase of the pro-apoptotic protein Bax, subsequently leading to the release of cytochrome c from mitochondria to the cytosol and activating caspase-9, which in turn stimulated effector caspase-7. Rusvinoxidase at a dose of 4 mg/kg was non-toxic in mice, indicating that it may be useful as a model for the development of peptide-based anticancer drugs. PMID:26319994

  2. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical synthesis of ceramide is a costly process, and developments of alternative cost......-efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through enzymatic hydrolysis of sphingomyelin have been studied. Sphingomyelin (SM) is a ubiquitous membrane-lipid and dairy products or by-products is a rich source of sphingomyelin. In...

  3. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Zhang, Long; Hellgren, Lars; Xu, Xuebing

    Sphingomyelin (SM) is a ubiquitous component of animal cell membranes, and it is the most abundant sphingolipid. Ceramide, a hydrolysis product from SM, has an important role in cellular signaling, and especially in the regulation of apoptosis, cell differentiation, transformation and proliferation....... Therefore, it is desirable to develop alternative cost-efficient, high yield production methods. This study optimized the enzymatic production of ceramide from SM. Phospholipase C from Clostridium perfringens was chosen to catalyze the reaction. Several important factors were considered in optimization....

  4. Enzymatic cascade bioreactor

    Science.gov (United States)

    Simmons, Blake A.; Volponi, Joanne V.; Ingersoll, David; Walker, Andrew

    2007-09-04

    Disclosed is an apparatus and method for continuously converting sucrose to .beta.-D-glucose. The method comprises a three stage enzymatic reactor in which an aqueous solution of sucrose is first converted into a solution of fructose and .alpha.-D-glucose by passing it through a porous, packed column containing an inert media on which invertase is immobilized. This solution is then sent through a second packed column containing glucose isomerase and finally a third packed column containing mutarotase. Solution temperature and pH are adjusted to maximize glucose output.

  5. Ultrasonic-assisted enzymatic extraction of silymarin from the Silybum marianum seed shell and evaluation of its antioxidant activity in vitro

    OpenAIRE

    Zhao, Fei; Li, Xinhua

    2015-01-01

    This study revealed the optimal conditions for the Ultrasonic-Assisted Enzymatic Extraction (UAEE) of silymarin, and include: the concentration of ethanol, 50 %; enzyme concentration, 30 U/mg; liquid-solid ratio, 6:1; an extraction time of 120 min; and the ultrasonic power at 180 W. The extraction rate was 7.86 %, which is higher, by 74.67 %, than that of the silymarin extract from the Silybum marianum meal prepared by a distinct approach. SEM micrographs of the inner and outer surfaces of th...

  6. Electrical activity of ferroelectric biomaterials and its effects on the adhesion, growth and enzymatic activity of human osteoblast-like cells

    Science.gov (United States)

    Vaněk, P.; Kolská, Z.; Luxbacher, T.; García, J. A. L.; Lehocký, M.; Vandrovcová, M.; Bačáková, L.; Petzelt, J.

    2016-05-01

    Ferroelectrics have been, among others, studied as electroactive implant materials. Previous investigations have indicated that such implants induce improved bone formation. If a ferroelectric is immersed in a liquid, an electric double layer and a diffusion layer are formed at the interface. This is decisive for protein adsorption and bioactive behaviour, particularly for the adhesion and growth of cells. The charge distribution can be characterized, in a simplified way, by the zeta potential. We measured the zeta potential in dependence on the surface polarity on poled ferroelectric single crystalline LiNbO3 plates. Both our results and recent results of colloidal probe microscopy indicate that the charge distribution at the surface can be influenced by the surface polarity of ferroelectrics under certain ‘ideal’ conditions (low ionic strength, non-contaminated surface, very low roughness). However, suggested ferroelectric coatings on the surface of implants are far from ideal: they are rough, polycrystalline, and the body fluid is complex and has high ionic strength. In real cases, it can therefore be expected that there is rather low influence of the sign of the surface polarity on the electric diffusion layer and thus on the specific adsorption of proteins. This is supported by our results from studies of the adhesion, growth and the activity of alkaline phosphatase of human osteoblast-like Saos-2 cells on ferroelectric LiNbO3 plates in vitro.

  7. Biofunctional Properties of Enzymatic Squid Meat Hydrolysate

    Science.gov (United States)

    Choi, Joon Hyuk; Kim, Kyung-Tae; Kim, Sang Moo

    2015-01-01

    Squid is one of the most important commercial fishes in the world and is mainly utilized or consumed as sliced raw fish or as processed products. The biofunctional activities of enzymatic squid meat hydrolysate were determined to develop value-added products. Enzymatic squid hydrolysate manufactured by Alcalase effectively quenched 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and hydrogen peroxide radical with IC50 values of 311, 3,410, and 111.5 μg/mL, respectively. Angiotensin I-converting enzyme inhibitory activity of squid hydrolysate was strong with an IC50 value of 145.1 μg/mL, while tyrosinase inhibitory activity with an IC50 value of 4.72 mg/mL was moderately low. Overall, squid meat hydrolysate can be used in food or cosmetic industries as a bioactive ingredient and possibly be used in the manufacture of seasoning, bread, noodle, or cosmetics. PMID:25866752

  8. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Camila Ramos; Meza, Andreia Navarro [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Giesel, Guilherme Menegon; Verli, Hugo [Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Squina, Fabio Marcio [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Prade, Rolf Alexander [Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK (United States); Murakami, Mario Tyago, E-mail: mario.murakami@lnbio.org.br [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil)

    2010-12-10

    Research highlights: {yields} The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. {yields} Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. {yields} Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 {sup o}C, and exclusively xylobiose at 90 {sup o}C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  9. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    International Nuclear Information System (INIS)

    Research highlights: → The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. → Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. → Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 oC, and exclusively xylobiose at 90 oC as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  10. Selective Enzymatic Reduction of Aldehydes

    Directory of Open Access Journals (Sweden)

    Patrizia Di Gennaro

    2006-05-01

    Full Text Available Highly selective enzymatic reductions of aldehydes to the corresponding alcohols was performed using an E. coli JM109 whole cell biocatalyst. A selective enzymatic method for the reduction of aldehydes could provide an eco-compatible alternative to chemical methods. The simplicity, fairly wide scope and the very high observed chemoselectivity of this approach are its most unique features.

  11. Enzymatic radioassay for gentamicin

    International Nuclear Information System (INIS)

    An enzymatic radiochemical assay procedure for measuring serum gentamicin by use of gentamicin 3-acetyltransferase and [acetyl-14C]acetyl-Coenzyme A is described and evaluated. The enzyme stoichiometrically and quantitatively transfers a radioactive label to the analyte during a 10-min incubation at 370C. The labeled gentamicin is then adsorbed onto phosphocellulose paper discs, which are washed to remove unreacted [acetyl-14C]acetyl-Coenzyme A and counted in a liquid scintillation counter for 1 min each. The assay detects gentamicin in concentrations as low as 0.2 mg/liter and gives a linear response to concentrations as high as 20 mg/liter. Sisomicin, a structural analog of gentamicin, is measured by the procedure, and tobramycin and netilmicin are slightly reactive. No other interferents were found among other aminoglycosides, other antibiotics, or substances endogenous to serum. Results by the new method are compared to those by radioimmunoassay and a microbiological method

  12. Specific enzymatic dephosphorylation of the retinoblastoma protein.

    OpenAIRE

    Ludlow, J W; Glendening, C L; Livingston, D M; DeCarprio, J A

    1993-01-01

    The retinoblastoma gene product (RB) undergoes cell cycle-dependent phosphorylation and dephosphorylation. Pulse-chase experiments revealed that the change in RB gel electrophoretic migration which occurs near mitosis is due to enzymatic dephosphorylation (J. W. Ludlow, J. Shon, J. M. Pipas, D. M. Livingston, and J. A. DeCaprio, Cell 60:387-396, 1990). To determine the precise timing of RB dephosphorylation and whether a specific phosphatase is active in this process, we have utilized a nocod...

  13. Screening of pumpkin cellulose degradation bacterium and its enzymatic activity determination%降解南瓜纤维素菌株的分离筛选及酶活测定

    Institute of Scientific and Technical Information of China (English)

    尚婷婷; 李思杨; 杨瑞学; 李全宏

    2012-01-01

    The objective of this article is to select the bacterial strains, which is capable of degrading pumpkin cellulose effectively, The experimental method is to extract the strains of degraded pumpkin cellulose from the soil, rotten leaves and fruit. From the beginning, transparent Congo red hydrolysis circles with the function of dyeing are operated to carry out the first selection, and next, CMC-Na, filter paper, pumpkin dregs are utilized to test for carbon source the enzymatic activity of carboxymethylcellulose from all the single and mixed bacterial strains. Ultimately, the strains with the strongest activity are taken as the representative to implement the activities on the next stage. The achievement of this experiment is to have chosen out five bacterial and seven fungi with the effective capacity of degrading cellulose. By means of the measurement of the enzymatic activity in the single and mixed strains, a result has been achieved that the enzymatic activity in mixed strains high three times than that of the single ones. Consequently, the conclusion is that the enzymatic activity in mixedstrains is much stronger than that of the single ones, fermentation cultivation 72 h pumpkin polysaccharide degradation soluble slag is the strongest force.%目的:筛选出能高效降解南瓜纤维素的菌株,以制备南瓜可溶性膳食纤维。方法:从土壤、腐烂的树叶和水果上分离出具有降解南瓜纤维素的茵株,用刚果红染色透明水解圈进行初筛,然后用CMC-Na、滤纸和南瓜渣为碳源测所有菌株及混合菌株的羧甲基纤维素酶活力,最终选出1株酶活力较高菌株进行下一步实验。结果:共分离到能够有效降解纤维素的共有5株细菌和7株真菌,通过测单菌株和混合菌株的酶活力,表明混合菌株的酶活力最大值比单菌株的酶活力最大值要高3倍之多。结论:混合菌株的酶活力比单株茵酶活力值高,发酵培养72h南瓜渣可溶性多糖降解力最强。

  14. Graphene based enzymatic bioelectrodes and biofuel cells

    Science.gov (United States)

    Karimi, Anahita; Othman, Ali; Uzunoglu, Aytekin; Stanciu, Lia; Andreescu, Silvana

    2015-04-01

    The excellent electrical conductivity and ease of functionalization make graphene a promising material for use in enzymatic bioelectrodes and biofuel cells. Enzyme based biofuel cells have attracted substantial interest due to their potential to harvest energy from organic materials. This review provides an overview of the functional properties and applications of graphene in the construction of biofuel cells as alternative power sources. The review covers the current state-of-the-art research in graphene based nanomaterials (physicochemical properties and surface functionalities), the role of these parameters in enhancing electron transfer, the stability and activity of immobilized enzymes, and how enhanced power density can be achieved. Specific examples of enzyme immobilization methods, enzyme loading, stability and function on graphene, functionalized graphene and graphene based nanocomposite materials are discussed along with their advantages and limitations. Finally, a critical evaluation of the performance of graphene based enzymatic biofuel cells, the current status, challenges and future research needs are provided.

  15. Homogeneous, Heterogeneous, and Enzymatic Catalysis.

    Science.gov (United States)

    Oyama, S. Ted; Somorjai, Gabor A.

    1988-01-01

    Discusses three areas of catalysis: homegeneous, heterogeneous, and enzymatic. Explains fundamentals and economic impact of catalysis. Lists and discusses common industrial catalysts. Provides a list of 107 references. (MVL)

  16. Enzymatic reactions in confined environments

    Science.gov (United States)

    Küchler, Andreas; Yoshimoto, Makoto; Luginbühl, Sandra; Mavelli, Fabio; Walde, Peter

    2016-05-01

    Within each biological cell, surface- and volume-confined enzymes control a highly complex network of chemical reactions. These reactions are efficient, timely, and spatially defined. Efforts to transfer such appealing features to in vitro systems have led to several successful examples of chemical reactions catalysed by isolated and immobilized enzymes. In most cases, these enzymes are either bound or adsorbed to an insoluble support, physically trapped in a macromolecular network, or encapsulated within compartments. Advanced applications of enzymatic cascade reactions with immobilized enzymes include enzymatic fuel cells and enzymatic nanoreactors, both for in vitro and possible in vivo applications. In this Review, we discuss some of the general principles of enzymatic reactions confined on surfaces, at interfaces, and inside small volumes. We also highlight the similarities and differences between the in vivo and in vitro cases and attempt to critically evaluate some of the necessary future steps to improve our fundamental understanding of these systems.

  17. Non-enzymatic browning reaction of glucosamine at mild conditions: Relationship between colour formation, radical scavenging activity and α-dicarbonyl compounds production.

    Science.gov (United States)

    Hong, Pui Khoon; Betti, Mirko

    2016-12-01

    Glucosamine (GlcN, 5% w/v) was incubated in either phosphate buffer or ammonium hydroxide solutions at 40 and 60°C for up to 48h in order to yield caramel solutions. Non-enzymatic browning was monitored via changes in absorption at 280, 320 and 420nm and the physico-chemical properties as well as the generation of short chain α-dicarbonyl compounds were evaluated. Accumulation of GlcN autocondensation products (280nm) proceeded in parallel with the development of pre-melanoidins (320nm) and melanoidins (420nm). The reactive α-dicarbonyls were detected at temperature as low as 40°C within 3h with a maximum level of diacetyl recorded at 6h. The caramel solutions showed a high efficacy in scavenging DPPH and ABTS radicals in accordance with the increasing browning intensity. The results suggest that GlcN browning can be modulated according to the specific desired properties to produce a multi-functional food ingredient that has health-promoting effects. PMID:27374528

  18. Enzymatic Synthesis of the Flavone Glucosides, Prunin and Isoquercetin, and the Aglycones, Naringenin and Quercetin, with Selective α-L-Rhamnosidase and β-D-Glucosidase Activities of Naringinase

    Directory of Open Access Journals (Sweden)

    Hélder Vila-Real

    2011-01-01

    Full Text Available The production of flavonoid glycosides by removing rhamnose from rutinosides can be accomplished through enzymatic catalysis. Naringinase is an enzyme complex, expressing both α-L-rhamnosidase and β-D-glucosidase activities, with application in glycosides hydrolysis. To produce monoglycosylated flavonoids with naringinase, the expression of β-D-glucosidase activity is not desirable leading to the need of expensive methods for α-L-rhamnosidase purification. Therefore, the main purpose of this study was the inactivation of β-D-glucosidase activity expressed by naringinase keeping α-L-rhamnosidase with a high retention activity. Response surface methodology (RSM was used to evaluate the effects of temperature and pH on β-D-glucosidase inactivation. A selective inactivation of β-D-glucosidase activity of naringinase was achieved at 81.5∘C and pH 3.9, keeping a very high residual activity of α-L-rhamnosidase (78%. This was a crucial achievement towards an easy and cheap production method of very expensive flavonoids, like prunin and isoquercetin starting from naringin and rutin, respectively.

  19. Identification of essential residues in 2',3'-cyclic nucleotide 3'-phosphodiesterase. Chemical modification and site-directed mutagenesis to investigate the role of cysteine and histidine residues in enzymatic activity.

    Science.gov (United States)

    Lee, J; Gravel, M; Gao, E; O'Neill, R C; Braun, P E

    2001-05-01

    2',3'-Cyclic nucleotide 3'-phosphodiesterase (CNP; EC ) catalyzes in vitro hydrolysis of 3'-phosphodiester bonds in 2',3'-cyclic nucleotides to produce 2'-nucleotides exclusively. N-terminal deletion mapping of the C-terminal two-thirds of recombinant rat CNP1 identified a region that possesses the catalytic domain, with further truncations abolishing activity. Proteolysis and kinetic analysis indicated that this domain forms a compact globular structure and contains all of the catalytically essential features. Subsequently, this catalytic fragment of CNP1 (CNP-CF) was used for chemical modification studies to identify amino acid residues essential for activity. 5,5'-Dithiobis-(2-nitrobenzoic acid) modification studies and kinetic analysis of cysteine CNP-CF mutants revealed the nonessential role of cysteines for enzymatic activity. On the other hand, modification studies with diethyl pyrocarbonate indicated that two histidines are essential for CNPase activity. Consequently, the only two conserved histidines, His-230 and His-309, were mutated to phenylalanine and leucine. All four histidine mutants had k(cat) values 1000-fold lower than wild-type CNP-CF, but K(m) values were similar. Circular dichroism studies demonstrated that the low catalytic activities of the histidine mutants were not due to gross changes in secondary structure. Taken together, these results demonstrate that both histidines assume critical roles for catalysis. PMID:11278504

  20. Enzymatic synthesis of spacer-linked divalent glycosides carrying N-acetylglucosamine and N-acetyllactosamine: analysis of cross-linking activities with WGA.

    Science.gov (United States)

    Misawa, Yoshinori; Akimoto, Takashi; Amarume, Satoshi; Murata, Takeomi; Usui, Taichi

    2008-01-01

    Divalent glycosides carrying N-acetyl-d-glucosamine (GlcNAc) and N-acetyllactosamine (LacNAc) were designed and prepared as glycomimetics. First, hexan-1,6-diyl bis-(2-acetamido-2-deoxy-beta-d-glucopyranoside) (GlcNAc-Hx-GlcNAc) and 3,6-dioxaoct-1,8-diyl bis-(2-acetamido-2-deoxy-beta-d-glucopyranoside) (GlcNAc-Doo-GlcNAc) were enzymatically synthesized by transglycosylation of an N,N'N'',N'''-tetraacetylchitotetraose [(GlcNAc)(4)] donor with a primary diol acceptor, utilizing a chitinolytic enzyme from Amycolatopsis orientalis. The resulting divalent glycosides were further converted to the respective hexan-1,6-diyl bis-[beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside] (LacNAc-Hx-LacNAc) and 6-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-hexyl beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside (LacNAc-Hx-GlcNAc), and respective 3,6-dioxaoct-1,8-diyl bis-[beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside] (LacNAc-Doo-LacNAc) and 8-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-3,6-dioxaoctyl beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside (LacNAc-Doo-GlcNAc) by galactosyltransferase. The interaction of wheat germ agglutinin (WGA) with a series of divalent glycosides and related compounds were studied using a biosensor based on surface plasmon resonance (SPR) and by precipitation analysis. Our results demonstrated that divalent glycosides carrying GlcNAc on both sides and GlcNAc and LacNAc on each side are capable of precipitating WGA as divalent ligands, but that the corresponding monovalent controls and divalent glycosides carrying LacNAc on both sides are unable to precipitate the lectin and bind as univalent ligands. PMID:17977857

  1. STABILIZATION OF MICROBIAL ENZYMATIC PREPARATIONS USED IN FEED INDUSTRY

    Directory of Open Access Journals (Sweden)

    MONICA DRAGOMIRESCU

    2013-12-01

    Full Text Available The enzymatic compound with protease activity obtained by Bacillus licheniformis CMIT 1.33 cells fermentation was entrapped in sol-gel matrixes, using TEOS as precursor. The sol-gel technique is a soft method for silica glass synthesis at room temperature. In silica gels, the enzymes maintain their catalytic activity for a longer period of time and present high stability at environmental factors. Bacillus licheniformis CMIT 1.33 enzyme was also immobilized by entrapment in silica gels deposited on a ceramic support. The optimal temperature and pH of the native and immobilized enzyme did not vary significantly. At temperature and pH values lower than the optimum, the relative activities have been higher for the immobilized compound. The immobilization of the enzymatic compound increased the stability. The enzymatic compounds with protease activity obtained by silica gels entrapment and entrapment/deposition on ceramic support can be used as forage additive.

  2. Alteração da atividade enzimática em organismos aquáticos por poluentes de origem agrícola: uma abordagem geral e sobre a suscetibilidade da fosfatase ácida Alteration of enzymatic activity in aquatic organisms by agricultural pollutants: a general approach and the susceptibility of the acid phosphatase

    Directory of Open Access Journals (Sweden)

    Claudio Martín Jonsson

    2010-01-01

    Full Text Available The input of agrochemicals in the aquatic compartment can results in biochemical injuries for living organisms. In this context, the knowledge of alterations of enzymatic activities due the presence of agriculture pollutants contributes for the elucidation of the mechanisms of toxicity, implementation of economic methods for monitoring purposes and establishment of maximum allowed concentrations. In the present work, the above considerations are discussed, and data concerning changes in enzymatic function by pesticides and fertilizer contaminants are reviewed. Also, we focused on the acid phosphatase due its susceptibility to several pollutants and diversity in cellular functions.

  3. Enzymatic and bacterial conversions during sourdough fermentation.

    Science.gov (United States)

    Gänzle, Michael G

    2014-02-01

    Enzymatic and microbial conversion of flour components during bread making determines bread quality. Metabolism of sourdough microbiota and the activity of cereal enzymes are interdependent. Acidification, oxygen consumption, and thiols accumulation by microbial metabolism modulate the activity of cereal enzymes. In turn, cereal enzymes provide substrates for bacterial growth. This review highlights the role of cereal enzymes and the metabolism of lactic acid bacteria in conversion of carbohydrates, proteins, phenolic compounds and lipids. Heterofermentative lactic acid bacteria prevailing in wheat and rye sourdoughs preferentially metabolise sucrose and maltose; the latter is released by cereal enzymes during fermentation. Sucrose supports formation of acetate by heterofermentative lactobacilli, and the formation of exopolysaccharides. The release of maltose and glucose by cereal enzymes during fermentation determines the exopolysaccharide yield in sourdough fermentations. Proteolysis is dependent on cereal proteases. Peptidase activities of sourdough lactic acid bacteria determine the accumulation of (bioactive) peptides, amino acids, and amino acid metabolites in dough and bread. Enzymatic conversion and microbial metabolism of phenolic compounds is relevant in sorghum and millet containing high levels of phenolic compounds. The presence of phenolic compounds with antimicrobial activity in sorghum selects for fermentation microbiota that are resistant to the phenolic compounds. PMID:24230468

  4. Enzymatic hydrolysis of polyester fabrics

    International Nuclear Information System (INIS)

    Enzymatic hydrolysis of polyester fabrics has been investigated, using different treatment times, temperature and concentration of enzymes. The effects of hydrolysis on samples were evaluated by measurement of weight loss, moisture regain, breaking load of warp yarns, thickness and Ftir spectroscopy. Results show that hydrolysis under mild conditions can improve moisture absorption of the samples. If the applied temperature, treatment time and concentration exceeded some specific range, the moisture regain would be affected negatively. The Ftir spectrums showed an increase in functional groups specially hydroxyl. However the effects of enzymatic hydrolysis on weight loss, tensile strength and thickness of polyester fabrics were negligible

  5. The dependence of the discharge of nitrous oxide by ordinary chernozem steppe of the Central-Chernozem Region of Russia from the content of humus, nitrogen and enzymatic activity

    Science.gov (United States)

    Avksentev, Alexey; Negrobova, Elena; Kramareva, Tatiana; Moiseeva, Evgenya

    2016-04-01

    The dependence of the discharge of nitrous oxide by ordinary chernozem steppe of the Central-Chernozem Region of Russia from the content of humus, nitrogen and enzymatic activity Alexey Avksentev, Elena Negrobova, Tatiana Kramareva, Evgenya Moiseeva 394000 Voronezh, Universitetskaya square, 1 Voronezh State University Nitrous oxide is emitted by soil as a result of microbiological processes, ranks third in the list of aggressive greenhouse gas after carbon dioxide and methane. Nitrous oxide is formed during nitrification and denitrification of ammonia that enters the soil during microbial decomposition of complex organic compounds. Denitrification can be direct and indirect. In the microbiological process of recovery of nitrates involved of the organic substance. In aerobic conditions microorganisms denitrificator behave like normal saprotrophs and oxidize organic matter in the act of breathing oxygen. Thus, they operate at different times two enzyme systems: the electron transport chain with an oxygen acceptor in aerobic and restoration of nitrates under anaerobic conditions. Investigation of the emission of nitrous oxide by ordinary Chernozem steppe of the Central-Chernozem Region showed that it depends on the type of cenosis and the content of available forms of nitrogen. Natural ecosystems emit nitrous oxide more than the soil of arable land. The dependence of the emission of nitrous oxide from the humus content shows positive trend, but the aggregation of data, significant differences are not detected. Research shows that nitrous oxide emissions are seasonal. So the autumn season is characterized by nitrous oxide emissions than spring. Enzymatic processes are an important link in the biological cycle of elements and, consequently, participate in the process of decomposition of organic matter, nitrification and other processes. Analysis of the data on enzyme activity of ordinary Chernozem and the intensity of emission of N20 shows a clear relationship between

  6. Enzymatic properties of the glycine D-alanine [corrected] aminopeptidase of Aspergillus oryzae and its activity profiles in liquid-cultured mycelia and solid-state rice culture (rice koji).

    Science.gov (United States)

    Marui, Junichiro; Matsushita-Morita, Mayumi; Tada, Sawaki; Hattori, Ryota; Suzuki, Satoshi; Amano, Hitoshi; Ishida, Hiroki; Yamagata, Youhei; Takeuchi, Michio; Kusumoto, Ken-Ichi

    2012-01-01

    The gdaA gene encoding S12 family glycine-D-alanine aminopeptidase (GdaA) was found in the industrial fungus Aspergillus oryzae. GdaA shares 43% amino acid sequence identity with the D-aminopeptidase of the Gram-negative bacterium Ochrobactrum anthropi. GdaA purified from an A. oryzae gdaA-overexpressing strain exhibited high D-stereospecificity and efficiently released N-terminal glycine and D-alanine of substrates in a highly specific manner. The optimum pH and temperature were 8 to 9 and 40°C, respectively. This enzyme was stable under alkaline conditions at pH 8 to 11 and relatively resistant to acidic conditions until pH 5.0. The chelating reagent EDTA, serine protease inhibitors such as AEBSF, benzamidine, TPCK, and TLCK, and the thiol enzyme inhibitor PCMB inhibited the enzyme. The aminopeptidase inhibitor bestatin did not affect the activity. GdaA was largely responsible for intracellular glycine and D-alanine aminopeptidase activities in A. oryzae during stationary-phase growth in liquid media. In addition, the activity increased in response to the depletion of nitrogen or carbon sources in the growth media, although the GdaA-independent glycine aminopeptidase activity highly increased simultaneously. Aminopeptidases of A. oryzae attract attention because the enzymatic release of a variety of amino acids and peptides is important for the enhancement of the palatability of fermented foods. GdaA activity was found in extracts of a solid-state rice culture of A. oryzae (rice koji), which is widely used as a starter culture for Japanese traditional fermented foods, and was largely responsible for the glycine and D-alanine aminopeptidase activity detected at a pH range of 6 to 9. PMID:22005737

  7. Enzymatic Processing of Bioactive Glycosides from Natural Sources

    Science.gov (United States)

    Weignerová, Lenka; Křen, Vladimír

    A number of biologically active natural products are glycosides. Often, the glycosidic residue is crucial for their activity. In other cases, glycosylation only improves their pharmacokinetic parameters. Enzymatic modification of these glycosides - both extension of the glycoside moiety and its selective trimming - is advantageous due to their selectivity and mildness of the reaction conditions in the presence of reactive and sensitive complex aglycones. Enzymatic reactions enable the resulting products to be used as "natural products", e.g., in nutraceuticals. This chapter concentrates on naturally occurring glycosides used in medicine but also in the food and flavor industry (e.g., sweeteners). Both "classical" and modern methods will be discussed.

  8. A Comparison between the Effects of Albendazole and Meben¬ dazole on the Enzymatic Activity of Excretory / Secretory Prod-ucts of Echinococcus granulosus Protoscoleces in Vitro

    Directory of Open Access Journals (Sweden)

    Seyed Jafar ADNANI SADATI

    2016-02-01

    Full Text Available Background: Hydatid cysts are formed in human body can be treated clinically by surgery or drugs such as albendazole (ABZ and mebendazole (MBZ. The purpose of this study was comparing the effects of ABZ and MBZ on glutathione-S-transferase, alkaline phosphatase and protease enzymes activities in protoscoleces of hydatid cyst. Methods: The culture supernatants containing the parasite Excretory / Secretory (E/S products were collected every 12 h for 72 h. The E/S products of treated samples with 1µg/ml ABZ and MBZ and the control one were collected and after centrifugation then protein concentrations were measured according to Bradford method. GST, ALP and protease activities of E/S products were assessed photometrically.Results: The mean of GST specific activity level in treated protoscoleces with ABZ and MBZ and in control group were obtained 69.44, 132.83 and 225.47U/mg/protein/ml respectively. The mean ALP activity level in treated protoscoleces with ABZ and MBZ and in control group were detected 19.22, 22.27 and 27.85 U/mg/protein/ml respectively. The protease activity level in treated protoscoleces with ABZ and MBZ were not detected. While the mean of protease activity level in control group was 7.61U/mg/proteins. Statistical analysis showed the significant difference between protein concentrations, the specific activities of GST, ALP and protease enzymes in treated protoscoleces in comparison with control group (P<0.05. Also, the significant difference were seen between specific activities of GST and ALP enzymes in treated protoscoleces with ABZ in comparison with treated group with MBZ (P<0.05.Conclusion: ABZ is more effective on the enzymes activities (GST and ALP as compared with MBZ. Keywords: Hydatid cyst protoscoleces, Albendazole, Mebandazole, Protease, Glutathione S-Transferase, Alkaline phosphatase

  9. Enhanced removal of detergent and recovery of enzymatic activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis: UUse of casein in gel wash buffer

    International Nuclear Information System (INIS)

    The inclusion of 1% casein or bovine serum albumin in buffer used to reactivate enzymes subjected to sodium dodecyl sulfate (SDS)-polyacrylamide electrophoresis resulted in accelerated removal of SDS and restoration of nuclease and beta-galactosidase enzyme activities. Nuclease and beta-galactosidase activities which are absent from gels after longer wash procedures are detectable with this technique. Enzyme activity in gels prepared with SDS which contained inhibitory contaminants was partially restored by the casein wash procedure. The threshold of detection of two-dimensionally separated deoxyribonuclease I using the casein wash procedure was 1 picogram

  10. Analysis on enzymatic browning in pine needles

    Energy Technology Data Exchange (ETDEWEB)

    Kong, K.H.; Park, H.J.; Choi, S.S.; Cho, S.H. [Chung-Ang University, Seoul (Korea); Kim, Y.T. [Aoyama Gakuin University, Tokyo (Japan)

    1999-06-01

    Tyrosinases are related to the enzymatic browning of plants and attract the major scientific interest for the prevention of it. Three tyrosinase isozymes (P{sub 1}, P{sub 2} and P{sub 3}) from pine needles were purified to homogeneity and characterized the factors that affect their activities. The L-ascorbic acid and {beta}-mercaptoethanol notably inhibited the enzymatic activities of the three isozymes. The sodium diethyldithiocarbamate was a competitive inhibitor of isozymes with the K{sub i} values of P{sub 1}(0.30 mM), P{sub 2}(0.015 mM) and P{sub 3}(0.019 mM), respectively. Their enzyme activities were however, increased by the addition of most metal ions. The optimum pH for the three isozymes was 9.0{approx}9.5 and the optimum temperatures ranged from 55 to 60{sup o} C using L-DOPA as substrate. 15 refs., 3 figs., 2 tabs.

  11. Inhibition of Class II Major Histocompatibility Complex Antigen Processing by Escherichia coli Heat-Labile Enterotoxin Requires an Enzymatically Active A Subunit

    Science.gov (United States)

    Matousek, Milita P.; Nedrud, John G.; Cieplak, Witold; Harding, Clifford V.

    1998-01-01

    Escherichia coli heat-labile enterotoxin (LT) and cholera toxin (CT) were found to inhibit intracellular antigen processing. Processing was not inhibited by mutant LT with attenuated ADP-ribosyltransferase activity, CT B or LT B subunit, which enhanced presentation of preexisting cell surface peptide-class II major histocompatibility complex complexes. Inhibition of antigen processing correlated with A subunit ADP-ribosyltransferase activity. PMID:9632629

  12. Bioluminescence methods for enzymatic determinations

    International Nuclear Information System (INIS)

    An enzymatic method for continuous, on-line and rapid detection of diagnostically useful biomarkers, which are symptomatic of disease or trauma-related tissue damage, is disclosed. The method is characterized by operability on authentic samples of complex biological fluids which contain the biomarkers

  13. Bioluminescence methods for enzymatic determinations

    Science.gov (United States)

    Bostick, William D.; Denton, Mark S.; Dinsmore, Stanley R.

    1982-01-01

    An enzymatic method for continuous, on-line and rapid detection of diagnostically useful biomarkers, which are symptomatic of disease or trauma-related tissue damage, is disclosed. The method is characterized by operability on authentic samples of complex biological fluids which contain the biomarkers.

  14. Imbalance in the enzymatic system of production and consumption of active oxygen species in liver of AKR mice with spontaneous leucosis.

    Science.gov (United States)

    Vartanyan, L S; Gurevich, S M; Kozachenko, A I; Nagler, L G; Burlakova, E B

    2001-07-01

    Activities of protective antioxidant enzymes, the rate of superoxide formation (v) in microsomal membranes and submitochondrial particles (SMP), and the concentrations of reduced and oxidized glutathione in cytosol were studied in the liver of AKR mice during the development of spontaneous leucosis. It was found that in the latent period of leucosis (mice of 3-6 months of age) the glutathione reductase (GR) activity in cytosol and mitochondria decreased and v in SMP increased. The increase in v in SMP did not result in the induction of Mn-SOD. In this stage of leucosis, the activities of Cu,Zn-SOD, GSH-Px, and G-6-PDH in cytosol were unchanged; at the same time, the GR activity and the concentration of reduced glutathione smoothly decreased. In the stage of developed leucosis (mice of 7-9 months of age), non-synchronous changes in the antioxidant system resulting in the shift of metabolism towards the prooxidant state were found. Comparison of our findings and the literature data demonstrates that the observed decrease in the SOD/GSH-Px ratio, the decrease in GR activity, and the increase in the v/Mn-SOD activity ratio are typical for pre-neoplastic changes in cell metabolism. PMID:11563951

  15. Enzymatic Activities of Human Cytomegalovirus Maturational Protease Assemblin and Its Precursor (pPR, pUL80a): Maximal Activity of pPR Requires Self-Interaction through Its Scaffolding Domain▿

    OpenAIRE

    Brignole, Edward J.; Gibson, Wade

    2007-01-01

    Herpesviruses encode an essential, maturational serine protease whose catalytic domain, assemblin (28 kDa), is released by self-cleavage from a 74-kDa precursor (pPR, pUL80a). Although there is considerable information about the structure and enzymatic characteristics of assemblin, a potential pharmacologic target, comparatively little is known about these features of the precursor. To begin studying pPR, we introduced five point mutations that stabilize it against self-cleavage at its intern...

  16. A correlation between the fate and non-extractable residue formation of 14C-metalaxyl and enzymatic activities in soil.

    Science.gov (United States)

    Botterweck, Jens; Claßen, Daniela; Zegarski, Thordis; Gottfroh, Christian; Kalathoor, Roshni; Schäffer, Andreas; Schwarzbauer, Jan; Schmidt, Burkhard

    2014-01-01

    Extracellular, oxidative soil enzymes like monophenol oxidases and peroxidases play an important role in transformation of xenobiotics and the formation of organic matter in soil. Additionally, these enzymes may be involved in the formation of non-extractable residues (NERs) of xenobiotics during humification processes. To examine this correlation, the fate of the fungicide (14)C metalaxyl in soil samples from Ultuna (Sweden) was studied. Using different soil sterilization techniques, it was possible to differentiate between free, immobilized, and abiotic ("pseudoenzyme"-like) oxidative activities. A correlation between the formation of metalaxyl NER and soil organic matter content, biotic activities, as well as extracellular phenoloxidase and peroxidase activities in the bulk soil and its particle size fractions was determined. Extracellular soil-bound enzymes were involved in NER formation (up to 8% of applied radioactivity after 92 days) of the fungicide independently from the presence of living microbes and different distributions of the NER in the soil humic subfractions. PMID:24328538

  17. Variation in metabolic enzymatic activity in white muscle and liver of blue tilapia, Oreochromis aureus, in response to long-term thermal acclimatization

    Science.gov (United States)

    Younis, Elsayed M.

    2015-05-01

    The effects of rearing temperature on white muscle and hepatic phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were examined in fingerlings of blue tilapia, Oreochromis aureus. The experiment was conducted for 14 weeks at temperatures of 18, 22, 26, 30, and 34°C. The activity of the glycolytic enzymes PFK, PK, and LDH in white muscle increased significantly with increase in water temperature. A reverse trend was observed for these enzymes in the liver, except for LDH, which behaved in the same manner as in white muscle. Cytosolic AST and ALT activity increased in both white muscle and liver in response to warm thermal acclimatization, while a reduction in mitochondrial AST and ALT activity was noticed at high temperatures in comparison with those at a lower temperature.

  18. The ubiquitin ligase HERC3 attenuates NF-κB-dependent transcription independently of its enzymatic activity by delivering the RelA subunit for degradation.

    Science.gov (United States)

    Hochrainer, Karin; Pejanovic, Nadja; Olaseun, Victoria A; Zhang, Sheng; Iadecola, Costantino; Anrather, Josef

    2015-11-16

    Activation of NF-κB-dependent transcription represents an important hallmark of inflammation. While the acute inflammatory response is per se beneficial, it can become deleterious if its spatial and temporal profile is not tightly controlled. Classically, NF-κB activity is limited by cytoplasmic retention of the NF-κB dimer through binding to inhibitory IκB proteins. However, increasing evidence suggests that NF-κB activity can also be efficiently contained by direct ubiquitination of NF-κB subunits. Here, we identify the HECT-domain ubiquitin ligase HERC3 as novel negative regulator of NF-κB activity. We find that HERC3 restricts NF-κB nuclear import and DNA binding without affecting IκBα degradation. Instead HERC3 indirectly binds to the NF-κB RelA subunit after liberation from IκBα inhibitor leading to its ubiquitination and protein destabilization. Remarkably, the regulation of RelA activity by HERC3 is independent of its inherent ubiquitin ligase activity. Rather, we show that HERC3 and RelA are part of a multi-protein complex containing the proteasome as well as the ubiquitin-like protein ubiquilin-1 (UBQLN1). We present evidence that HERC3 and UBQLN1 provide a link between NF-κB RelA and the 26S proteasome, thereby facilitating RelA protein degradation. Our findings establish HERC3 as novel candidate regulating the inflammatory response initiated by NF-κB. PMID:26476452

  19. High-Pressure-Assisted Enzymatic Release of Peptides and Phenolics Increases Angiotensin Converting Enzyme I Inhibitory and Antioxidant Activities of Pinto Bean Hydrolysates.

    Science.gov (United States)

    Garcia-Mora, Patricia; Peñas, Elena; Frias, Juana; Zieliński, Henryk; Wiczkowski, Wiesław; Zielińska, Danuta; Martínez-Villaluenga, Cristina

    2016-03-01

    Pinto bean protein concentrate was hydrolyzed by subtilisins at 0.1, 100, and 200 MPa and 50 °C for 15 min. Alcalase hydrolysis at 100 MPa led to higher ACE inhibition, reducing power, and free radical scavenging activity of hydrolysates. However, hydrolysate obtained by Savinase at 200 MPa showed the best ACE-inhibitory and radical scavenging activities. Proteolysis by Savinase at 200 MPa was considered the most effective treatment to increase small peptides (benefits in the production of functional hydrolysates providing higher functionality and added value to the resulting hydrolysate due to synergistic effects of bioactive peptides and soluble phenolics. PMID:26857428

  20. AKTIVITAS DAN STABILITAS RADICAL SCAVENGING L-ASKORBIL PALMITAT HASIL SINTESIS SECARA ENZIMATIK [Activity and Stability of Radical Scavenging of L- Palmitate Synthesized Enzymatically

    Directory of Open Access Journals (Sweden)

    Tri Agus Siswoyo1*

    2009-12-01

    Full Text Available L-ascorbyl palmitate (AsA-Pal-Enz was synthesized by using an immobilized lipase from Aspergillus niger. A comparison of antioxidative effects between L-ascorbic acid (AsA and AsA-Pal-Enz was determined in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH radical–scavenging. The results indicate that the AsA-Pal-Enz was effective in preventing lipid oxidation, while the antioxidative activity in authentic AsA-Pal was lower. The activity of AsA-Pal-Enz was very stable than AsA-Pal standard during heating.

  1. AKTIVITAS DAN STABILITAS RADICAL SCAVENGING L-ASKORBIL PALMITAT HASIL SINTESIS SECARA ENZIMATIK [Activity and Stability of Radical Scavenging of L- Palmitate Synthesized Enzymatically

    OpenAIRE

    Tri Agus Siswoyo; Tri Ardiyati 2)

    2009-01-01

    L-ascorbyl palmitate (AsA-Pal-Enz) was synthesized by using an immobilized lipase from Aspergillus niger. A comparison of antioxidative effects between L-ascorbic acid (AsA) and AsA-Pal-Enz was determined in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical–scavenging. The results indicate that the AsA-Pal-Enz was effective in preventing lipid oxidation, while the antioxidative activity in authentic AsA-Pal was lower. The activity of AsA-Pal-Enz was very stable than AsA-Pal standard duri...

  2. Enzymatic Oxidation of Methane

    Energy Technology Data Exchange (ETDEWEB)

    Sirajuddin, S; Rosenzweig, AC

    2015-04-14

    Methane monooxygenases (MMOs) are enzymes that catalyze the oxidation of methane to methanol in methanotrophic bacteria. As potential targets for new gas-to-liquid methane bioconversion processes, MMOs have attracted intense attention in recent years. There are two distinct types of MMO, a soluble, cytoplasmic MMO (sMMO) and a membrane-bound, particulate MMO (pMMO). Both oxidize methane at metal centers within a complex, multisubunit scaffold, but the structures, active sites, and chemical mechanisms are completely different. This Current Topic review article focuses on the overall architectures, active site structures, substrate reactivities, proteinprotein interactions, and chemical mechanisms of both MMOs, with an emphasis on fundamental aspects. In addition, recent advances, including new details of interactions between the sMMO components, characterization of sMMO intermediates, and progress toward understanding the pMMO metal centers are highlighted. The work summarized here provides a guide for those interested in exploiting MMOs for biotechnological applications.

  3. A versatile biosensing system for DNA-related enzyme activity assay via the synthesis of silver nanoclusters using enzymatically-generated DNA as template.

    Science.gov (United States)

    Yuan, Yijia; Li, Wenhua; Liu, Zhuoliang; Nie, Zhou; Huang, Yan; Yao, Shouzhuo

    2014-11-15

    In the present day, oligonucleotide-encapsulated silver clusters (DNA-AgNCs) have been widely applied into bio-analysis as a signal producer. Herein, we developed a novel method to synthesize DNA-AgNCs encapsulated by long-chain cytosine (C)-rich DNA. Such DNA was polymerized in a template-free way by terminal deoxynucleotidyl transferase (TdT). We demonstrated that TdT-polymerized long chain C-rich DNA can serve as an excellent template for AgNCs synthesis. Based on this novel synthesis strategy, we developed a label-free and turn-on fluorescence assay to detect TdT activity with ultralow limit of detection (LOD) of 0.0318 U and ultrahigh signal to background (S/B) of 46.7. Furthermore, our proposed method was extended to a versatile biosensing strategy for turn-on nucleases activity assay based on the enzyme-activated TdT polymerization. Two nucleases, EcoRI and ExoIII as model of endonuclease and exonuclease, respectively, have been detected with high selectivity and competitive low LOD of 0.0629 U and 0.00867 U, respectively. Our work demonstrates the feasibility of TdT polymerization-based DNA-AgNCs synthesis strategy as a versatile and potent biosensing platform to detect the activity of DNA-related enzymes. PMID:24907540

  4. Soil microbiological properties and enzymatic activities of long-term post-fire recovery in dry and semiarid Aleppo pine (Pinus halepensis M. forest stands

    Directory of Open Access Journals (Sweden)

    J. Hedo

    2014-10-01

    Full Text Available Wildfires affecting forest ecosystems and post-fire silvicultural treatments may cause considerable changes in soil properties. The capacity of different microbial groups to recolonize soil after disturbances is crucial for proper soil functioning. The aim of this work was to investigate some microbial soil properties and enzyme activities in semiarid and dry Aleppo pine (Pinus halepensis M. forest stands. Different plots affected by a wildfire event 17 years ago without or with post-fire silvicultural treatments five years after the fire event were selected. A mature Aleppo pine stand unaffected by wildfire and not thinned was used as a control. Physicochemical soil properties (soil texture, pH, carbonates, organic matter, electrical conductivity, total N and P, soil enzymes (urease, phosphatase, β-glucosidase and dehydrogenase activities, soil respiration and soil microbial biomass carbon were analysed in the selected forests areas and plots. The main finding was that long time after this fire event produces no differences in the microbiological soil properties and enzyme activities of soil after comparing burned and thinned, burned and not thinned, and mature plots. Thus, the long-term consequences and post-fire silvicultural management in the form of thinning have a significant effect on the site recovery after fire. Moreover, significant site variation was generally seen in soil enzyme activities and microbiological parameters. We conclude that total vegetation restoration normalises microbial parameters, and that wildfire and post-fire silvicultural treatments are not significant factors of soil properties after 17 years.

  5. Soil microbiological properties and enzymatic activities of long-term post-fire recovery in dry and semiarid Aleppo pine (Pinus halepensis M.) forest stands

    Science.gov (United States)

    Hedo, J.; Lucas-Borja, M. E.; Wic, C.; Andrés-Abellán, M.; de Las Heras, J.

    2015-02-01

    Wildfires affecting forest ecosystems and post-fire silvicultural treatments may cause considerable changes in soil properties. The capacity of different microbial groups to recolonise soil after disturbances is crucial for proper soil functioning. The aim of this work was to investigate some microbial soil properties and enzyme activities in semiarid and dry Aleppo pine (Pinus halepensis M.) forest stands. Different plots affected by a wildfire event 17 years ago without or with post-fire silvicultural treatments 5 years after the fire event were selected. A mature Aleppo pine stand, unaffected by wildfire and not thinned was used as a control. Physicochemical soil properties (soil texture, pH, carbonates, organic matter, electrical conductivity, total N and P), soil enzymes (urease, phosphatase, β-glucosidase and dehydrogenase activities), soil respiration and soil microbial biomass carbon were analysed in the selected forests areas and plots. The main finding was that long time after this fire event produces no differences in the microbiological soil properties and enzyme activities of soil after comparing burned and thinned, burned and not thinned, and mature plots. Moreover, significant site variation was generally seen in soil enzyme activities and microbiological parameters. We conclude that total vegetation recovery normalises post-fire soil microbial parameters, and that wildfire and post-fire silvicultural treatments are not significant factors affecting soil properties after 17 years.

  6. Small-Angle Neutron Scattering Reveals pH-Dependent Conformational Changes in Trichoderma reesei Cellobiohydrolase I: Implications for Enzymatic Activity

    Energy Technology Data Exchange (ETDEWEB)

    Pingali, Sai Venkatesh [ORNL; O' Neill, Hugh Michael [ORNL; McGaughey, Joseph [ORNL; Urban, Volker S [ORNL; Rempe, Caroline S [ORNL; Petridis, Loukas [ORNL; Smith, Jeremy C [ORNL; Evans, Barbara R [ORNL; Heller, William T [ORNL

    2011-01-01

    Cellobiohydrolase I (Cel7A) of the fungus Trichoderma reesei (now classified as an anamorph of Hypocrea jecorina) hydrolyzes crystalline cellulose to soluble sugars, making it of key interest for producing fermentable sugars from biomass for biofuel production. The activity of the enzyme is pH-dependent, with its highest activity occurring at pH 4 5. To probe the response of the solution structure of Cel7A to changes in pH, we measured small angle neutron scattering of it in a series of solutions having pH values of 7.0, 6.0, 5.3, and 4.2. As the pH decreases from 7.0 to 5.3, the enzyme structure remains well defined, possessing a spatial differentiation between the cellulose binding domain and the catalytic core that only changes subtly. At pH 4.2, the solution conformation of the enzyme changes to a structure that is intermediate between a properly folded enzyme and a denatured, unfolded state, yet the secondary structure of the enzyme is essentially unaltered. The results indicate that at the pH of optimal activity, the catalytic core of the enzyme adopts a structure in which the compact packing typical of a fully folded polypeptide chain is disrupted and suggest that the increased range of structures afforded by this disordered state plays an important role in the increased activity of Cel7A through conformational selection.

  7. Mapping the Reaction Coordinates of Enzymatic Defluorination

    Energy Technology Data Exchange (ETDEWEB)

    Chan, Peter W.Y.; Yakunin, Alexander F.; Edwards, Elizabeth A.; Pai, Emil F. (Toronto)

    2011-09-28

    The carbon-fluorine bond is the strongest covalent bond in organic chemistry, yet fluoroacetate dehalogenases can readily hydrolyze this bond under mild physiological conditions. Elucidating the molecular basis of this rare biocatalytic activity will provide the fundamental chemical insights into how this formidable feat is achieved. Here, we present a series of high-resolution (1.15-1.80 {angstrom}) crystal structures of a fluoroacetate dehalogenase, capturing snapshots along the defluorination reaction: the free enzyme, enzyme-fluoroacetate Michaelis complex, glycolyl-enzyme covalent intermediate, and enzyme-product complex. We demonstrate that enzymatic defluorination requires a halide pocket that not only supplies three hydrogen bonds to stabilize the fluoride ion but also is finely tailored for the smaller fluorine halogen atom to establish selectivity toward fluorinated substrates. We have further uncovered dynamics near the active site which may play pivotal roles in enzymatic defluorination. These findings may ultimately lead to the development of novel defluorinases that will enable the biotransformation of more complex fluorinated organic compounds, which in turn will assist the synthesis, detoxification, biodegradation, disposal, recycling, and regulatory strategies for the growing markets of organofluorines across major industrial sectors.

  8. Monitoring of Enzymatic Proteolysis Using Self-Assembled Quantum Dot-Protein Substrate Sensors

    Directory of Open Access Journals (Sweden)

    Aaron R. Clapp

    2008-07-01

    Full Text Available We have previously utilized hybrid semiconductor quantum dot- (QD- peptide substrates for monitoring of enzymatic proteolysis. In this report, we expand on this sensing strategy to further monitor protein-protease interactions. We utilize QDs self-assembled with multiple copies of dye-labeled proteins as substrates for the sensing of protease activity. Detection of proteolysis is based on changes in the rate of fluorescence resonance energy transfer (FRET between the QDs and the proximal dye-labeled proteins following protein digestion by added enzyme. Our study focused on two representative proteolytic enzymes: the cysteine protease papain and the serine protease endoproteinase K. Analysis of the enzymatic digestion allowed us to estimate minimal values for the enzymatic activities of each enzyme used. Mechanisms of enzymatic inhibition were also inferred from the FRET data collected in the presence of inhibitors. Potential applications of this technology include drug discovery assays and in vivo cellular monitoring of enzymatic activity.

  9. Avaliação do tempo de secagem e da atividade de óxido-redutases de yacon (Smallanthus sonchifolius sob tratamento químico Drying evaluation time and yacon (Smallanthus sonchifolius enzymatic activity inhibition under chemical treatment

    Directory of Open Access Journals (Sweden)

    Vivianne Montarroyos Padilha

    2009-10-01

    project aimed to evaluate the use of chemical agents in yacon processing to obtain flour in a way that inhibits enzymatic darkening of the product besides determining the enzymatic activity in these treatments. Samples of yacon without chemical inhibitions, yacon treated with 1.0g 100g-1 calcium chloride for 30 seconds and yacon treated with 0.5g 100g-1 potassium metabisulfite for 5 minutes were dried at 55oC in a ventilated greenhouse and the proportions of humidity and drying curves were determined. The peroxidase activities and polyphenol oxidase enzymes were checked before and after being dried with an enzymatic darkening possible biochemist marker of this tubercle. Regarding humidity Parameter all the three treatment were equivalent, but treatment 2 (calcium chloride reduced the humidity in lower time. Before and after the thermal treatment the enzymatic activity was higher in treatment 3 (potassium metabisulfite. The thermal action did not inhibit completely polyphenol oxidase and peroxidase. The treatment with calcium chloride at 1.0g 100g-1 for 30 minutes to obtain yacon flour was the one with better result despite the fact that it did not inhibit completely peroxidase and polyphenol oxidase enzymes activity, offering a better drying time better material of row firmness , facilitating the process for obtaining the meal.

  10. 同时具有光敏感性和酶催化功能的复合敏感膜%Both light sensitivity and enzymatic activity of the composite sensitive film

    Institute of Scientific and Technical Information of China (English)

    王海; 黄俊; 王超

    2011-01-01

    采用溶液浇铸法成膜,把荧光指示剂和葡萄糖氧化物酶(GOD)同时固定于醋酸纤维素膜上,得到同时具有光敏感性和酶催化能力的复合敏感膜.利用SEM和紫外-可见光分光光度计对复合敏感膜表面形貌和酶活性进行了分析.荧光指示剂没有泄漏,固定化酶的稳定性高于游离酶,表明此醋酸纤维素膜可作为一种优良的荧光指示剂和酶固定化载体.%Cellulose acetate film was prepared by solution casting method. Composite sensing film can be obtained via immobilization of fluorescent indicator and glucose oxidase on cellulose acetate film. Surface morphology of composite sensitive film and enzymatic activity were analyzed by scanning electron microscope and UV-visible spectrometer. Fluorescent indicator does not leak. The stability of the immobilized enzyme was higher than the free enzyme. All above indicate this kind of cellulose acetate film can be a good immobilization carrier of fluorescent indicator and enzyme.

  11. Enzymatic and toxinological activities of Hypnale hypnale (hump-nosed pit viper venom and its fractionation by ion exchange high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    CH Tan

    2011-01-01

    Full Text Available Hypnale hypnale (hump-nosed pit viper has been recently identified as one of the medically important venomous snakes in Sri Lanka and on the southwestern coast of India. The characterization of its venom is essential for understanding the pathophysiology of envenomation and for optimizing its management. In the present study, the biological properties of Hypnale hypnale venom and venom fractions obtained using Resource Q ion exchange chromatography were determined. The venom exhibited toxic activities typical of pit viper venom, comparable to that of its sister taxon, the Malayan pit viper (Calloselasma rhodostoma. Particularly noteworthy were its high activities of thrombin-like enzyme, proteases, phospholipase A2, L-amino acid oxidase and hyaluronidase. The thrombin-like enzyme was mainly acidic and distributed over several chromatography fractions, indicating its existence in multiple isoforms. The hemorrhagic and necrotic activities of the venom were likely associated with the proteolytic enzyme found mainly in the basic fraction. Phospholipase A2 and phosphomonoesterase exist in both acidic and basic isoforms, while L-amino acid oxidase and hyaluronidase are highly acidic. The venom clotting activity on fibrinogens showed distinct species specificity in the following increasing order for clotting time: bovine < rabbit < goat < human < horse < < dog, and was comparable to that of C. rhodostoma venom. Its clot formation on human fibrinogen is gradual and prolonged, a phenomenon suggestive of consumptive coagulopathy as a complication observed clinically. At an intramuscular sublethal dose, the venom did not cause acute kidney injury in a rodent model, contrary to the positive control group treated with Daboia russelii venom. Nephrotoxicity may result from higher venom doses in the context of coagulopathy, as a complication provoked by venom hematoxicity.

  12. Long-Term Reduction of Cocaine Self-Administration in Rats Treated with Adenoviral Vector-Delivered Cocaine Hydrolase: Evidence for Enzymatic Activity

    OpenAIRE

    Zlebnik, Natalie E.; Brimijoin, Stephen; Gao, Yang; Saykao, Amy T.; Parks, Robin J.; Carroll, Marilyn E.

    2014-01-01

    A new pharmacokinetic approach treating cocaine addiction involves rapidly metabolizing cocaine before it reaches brain reward centers using mutated human butyrylcholinesterase (BChE) or cocaine hydrolase (CocH). Recent work has shown that helper-dependent adenoviral (hdAD) vector-mediated plasma CocH reduced the locomotor-activating effects of cocaine and prevented reinstatement of cocaine-seeking behavior up to 6 months in rats. The present study investigated whether hdAD-CocH could decreas...

  13. Identification of an Iron-Sulfur Cluster That Modulates the Enzymatic Activity in NarE, a Neisseria meningitidis ADP-ribosyltransferase*

    OpenAIRE

    Del Vecchio, Mariangela; Pogni, Rebecca; Baratto, Maria Camilla; Nobbs, Angela; Rappuoli, Rino; Pizza, Mariagrazia; Balducci, Enrico

    2009-01-01

    In prokaryotes, mono-ADP-ribose transfer enzymes represent a family of exotoxins that display activity in a variety of bacterial pathogens responsible for causing disease in plants and animals, including those affecting mankind, such as diphtheria, cholera, and whooping cough. We report here that NarE, a putative ADP-ribosylating toxin previously identified from Neisseria meningitidis, which shares structural homologies with Escherichia coli heat labile enterotoxin and toxin from Vibrio chole...

  14. Polyphenolic content and antioxidant properties of Moringa oleifera leaf extracts and enzymatic activity of liver from goats supplemented with Moringa oleifera leaves/sunflower seed cake.

    Science.gov (United States)

    Moyo, B; Oyedemi, S; Masika, P J; Muchenje, V

    2012-08-01

    The study investigated antioxidant potency of Moringa oleifera leaves in different in vitro systems using standard phytochemical methods. The antioxidative effect on the activities of superoxide dismutase (SOD), catalase (CAT), lipid peroxidation (LPO) and reduced glutathione (GSH) were investigated in goats supplemented with M. oleifera (MOL) or sunflower seed cake (SC). The acetone extract had higher concentrations of total flavonoids (295.01 ± 1.89 QE/g) followed by flavonols (132.74 ± 0.83 QE/g), phenolics (120.33 ± 0.76 TE/g) and then proanthocyanidins (32.59 ± 0.50 CE/g) than the aqueous extract. The reducing power of both solvent extracts showed strong antioxidant activity in a concentration dependent manner. The acetone extract depicted higher percentage inhibition against DPPH, ABTS and nitric oxide radicals which were comparable with reference standard antioxidants (vitamin C and BHT). MOL increased the antioxidant activity of GSH (186%), SOD (97.8%) and catalase (0.177%). Lipid peroxidation was significantly reduced by MOL. The present study suggests that M. oleifera could be a potential source of compounds with strong antioxidant potential. PMID:22465510

  15. Production Response and Digestive Enzymatic Activity of the Pacific White Shrimp Litopenaeus vannamei (Boone, 1931 Intensively Pregrown in Microbial Heterotrophic and Autotrophic-Based Systems

    Directory of Open Access Journals (Sweden)

    Manuel J. Becerra-Dórame

    2012-01-01

    Full Text Available Shrimp postlarvae were reared into different microcosm systems without water exchange; a traditional system based on simple fertilization to improve microalgae concentration (control, an autotrophic system (AS based on the promotion of biofloc and biofilm by the addition of fertilizer and artificial substrates and a heterotrophic system (HS based on the promotion of heterotrophic bacteria by the addition of nitrogenous and carbonaceous sources and artificial substrates. Better growth performance and survival were registered in shrimp from the AS and HS compared to the control. Feed conversion ratios were below 0.7 for all treatments, but AS and HS were significantly lower than the control. Regarding digestive performance, no significant differences were observed for trypsin, amylase and lipase activities among AS and control shrimp; however, shrimp from HS showed a higher trypsin and amylase activities, suggesting a higher digestive activity caused by the presence of microbial bioflocs. The presence of biofilm and bioflocs composed by either autotrophic or heterotrophic organisms in combination with formulated feed improved the growth performance and survival of shrimp. Apparently, such combination fits the nutritional requirements of shrimp.

  16. A novel familial mutation in the PCSK1 gene that alters the oxyanion hole residue of proprotein convertase 1/3 and impairs its enzymatic activity.

    Directory of Open Access Journals (Sweden)

    Michael Wilschanski

    Full Text Available Four siblings presented with congenital diarrhea and various endocrinopathies. Exome sequencing and homozygosity mapping identified five regions, comprising 337 protein-coding genes that were shared by three affected siblings. Exome sequencing identified a novel homozygous N309K mutation in the proprotein convertase subtilisin/kexin type 1 (PCSK1 gene, encoding the neuroendocrine convertase 1 precursor (PC1/3 which was recently reported as a cause of Congenital Diarrhea Disorder (CDD. The PCSK1 mutation affected the oxyanion hole transition state-stabilizing amino acid within the active site, which is critical for appropriate proprotein maturation and enzyme activity. Unexpectedly, the N309K mutant protein exhibited normal, though slowed, prodomain removal and was secreted from both HEK293 and Neuro2A cells. However, the secreted enzyme showed no catalytic activity, and was not processed into the 66 kDa form. We conclude that the N309K enzyme is able to cleave its own propeptide but is catalytically inert against in trans substrates, and that this variant accounts for the enteric and systemic endocrinopathies seen in this large consanguineous kindred.

  17. Enzymatic oxidation of methane.

    Science.gov (United States)

    Sirajuddin, Sarah; Rosenzweig, Amy C

    2015-04-14

    Methane monooxygenases (MMOs) are enzymes that catalyze the oxidation of methane to methanol in methanotrophic bacteria. As potential targets for new gas-to-liquid methane bioconversion processes, MMOs have attracted intense attention in recent years. There are two distinct types of MMO, a soluble, cytoplasmic MMO (sMMO) and a membrane-bound, particulate MMO (pMMO). Both oxidize methane at metal centers within a complex, multisubunit scaffold, but the structures, active sites, and chemical mechanisms are completely different. This Current Topic review article focuses on the overall architectures, active site structures, substrate reactivities, protein-protein interactions, and chemical mechanisms of both MMOs, with an emphasis on fundamental aspects. In addition, recent advances, including new details of interactions between the sMMO components, characterization of sMMO intermediates, and progress toward understanding the pMMO metal centers are highlighted. The work summarized here provides a guide for those interested in exploiting MMOs for biotechnological applications. PMID:25806595

  18. EDTA 对镉胁迫下小白菜幼苗保护酶活性的影响%Effects of EDTA on the Protective Enzymatic Activities of Brassica Campestris L.chinensis under Cadmium Stress

    Institute of Scientific and Technical Information of China (English)

    赵辉; 郝振萍

    2015-01-01

    以3种小白菜为试验材料,探讨了乙二胺四乙酸(EDTA)对镉胁迫下小白菜幼苗保护酶活性的影响。结果表明:外施一定浓度的 EDTA 后,可以缓解镉胁迫对小白菜造成的伤害;当 EDTA 浓度为7.5 mmol·L-1时,小白菜 SOD、POD 酶活性最高,MDA 含量最低;当 EDTA 浓度为5 mmol·L-1时,CAT 酶活性最高;不同品种抗性表现不同,供试的3种小白菜品种中,抗热605的抗性最强。%Three species of Brassica campestris L.chinesis were used in this study to learn the effects of EDTA on the protective enzymatic activities of seedling under cadmium stress.The results showed that EDTA reduced the hurt from cadmium stress under special concentration. When the EDTA concentration was 7.5 mmol·L-1 ,the activities of SOD and POD were at their highest level and the content of MDA was the lowest.When the EDTA concentration was 5 mmol·L-1 ,the activity of CAT was the highest.Different species showed different reactions to cadmium stress,among which kangre 605 was the best during three species.

  19. 冰温贮藏对绿芦笋品质及酶活性的影响%Effect of ice-temperature preservation on quality and enzymatic activity of Green Asparagus

    Institute of Scientific and Technical Information of China (English)

    宋秀香; 鲁晓翔; 陈绍慧; 李江阔

    2013-01-01

    To 4℃ refrigerated storage for comparison,the effect of ice-temperature preservation on quality and enzymatic activity of Green Asparagus was investigated.The results showed that the storage life of Green asparagus was increased 14d,and its organoleptic quality was improved by ice-temperature preservation.While the firmness and tincture of Green asparagus was kept,the decrease of the V c content was slowed down,and its commodity value was improved by ice-temperature preservation.The activity of PPO and POD of Green asparagus was increased,and the activity of PAL was decreased under ice-temperature.So the protection ability of Green asparagus was enhanced,and the aging of Green asparagus was delayed.The control effect of ice-temperature preservation on respiration and ethylene production was unconspicuous.%以4℃冷藏作为对照,研究了冰温贮藏对绿芦笋品质及酶活性的影响.结果表明:冰温贮藏延长了绿芦笋贮藏期14d,提高其感官品质;同时,冰温贮藏保持了绿芦笋的硬度和色泽,减缓了Vc含量的降低,提高了其商品价值;冰温条件下绿芦笋多酚氧化酶(PPO)和过氧化物酶(POD)的活性提高,苯丙氨酸解氨酶(PAL)活性降低,增强了自身保护能力,延缓绿芦笋衰老;但冰温贮藏对绿芦笋呼吸作用和乙烯生成量的抑制效果不明显.

  20. Crystal Structure of Full-length Mycobacterium tuberculosis H37Rv Glycogen Branching Enzyme; Insights of N-Terminal [beta]-Sandwich in Sustrate Specifity and Enzymatic Activity

    Energy Technology Data Exchange (ETDEWEB)

    Pal, Kuntal; Kumar, Shiva; Sharma, Shikha; Garg, Saurabh Kumar; Alam, Mohammad Suhail; Xu, H. Eric; Agrawal, Pushpa; Swaminathan, Kunchithapadam (NU Sinapore); (Van Andel); (IMT-India)

    2010-07-13

    The open reading frame Rv1326c of Mycobacterium tuberculosis (Mtb) H37Rv encodes for an {alpha}-1,4-glucan branching enzyme (MtbGlgB, EC 2.4.1.18, Uniprot entry Q10625). This enzyme belongs to glycoside hydrolase (GH) family 13 and catalyzes the branching of a linear glucose chain during glycogenesis by cleaving a 1 {yields} 4 bond and making a new 1 {yields} 6 bond. Here, we show the crystal structure of full-length MtbGlgB (MtbGlgBWT) at 2.33-{angstrom} resolution. MtbGlgBWT contains four domains: N1 {beta}-sandwich, N2 {beta}-sandwich, a central ({beta}/{alpha}){sub 8} domain that houses the catalytic site, and a C-terminal {beta}-sandwich. We have assayed the amylase activity with amylose and starch as substrates and the glycogen branching activity using amylose as a substrate for MtbGlgBWT and the N1 domain-deleted (the first 108 residues deleted) Mtb{Delta}108GlgB protein. The N1 {beta}-sandwich, which is formed by the first 105 amino acids and superimposes well with the N2 {beta}-sandwich, is shown to have an influence in substrate binding in the amylase assay. Also, we have checked and shown that several GH13 family inhibitors are ineffective against MtbGlgBWT and Mtb{Delta}108GlgB. We propose a two-step reaction mechanism, for the amylase activity (1 {yields} 4 bond breakage) and isomerization (1 {yields} 6 bond formation), which occurs in the same catalytic pocket. The structural and functional properties of MtbGlgB and Mtb{Delta}108GlgB are compared with those of the N-terminal 112-amino acid-deleted Escherichia coli GlgB (EC{Delta}112GlgB).

  1. The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme: GLYCOSYLATION IS REQUIRED FOR ENZYMATIC ACTIVITY BUT NOT FOR OLIGOMERIZATION*

    OpenAIRE

    Chen, Vicky P.; Choi, Roy C. Y.; Chan, Wallace K. B.; Leung, K. Wing; Guo, Ava J. Y.; Gallant K. L. Chan; Luk, Wilson K. W.; Tsim, Karl W. K.

    2011-01-01

    Acetylcholinesterase (AChE) anchors onto cell membranes by a transmembrane protein PRiMA (proline-rich membrane anchor) as a tetrameric form in vertebrate brain. The assembly of AChE tetramer with PRiMA requires the C-terminal “t-peptide” in AChE catalytic subunit (AChET). Although mature AChE is well known N-glycosylated, the role of glycosylation in forming the physiologically active PRiMA-linked AChE tetramer has not been studied. Here, several lines of evidence indicate that the N-linked ...

  2. Phytase production by Rhizopus microsporus var. microsporus biofilm: characterization of enzymatic activity after spray drying in presence of carbohydrates and nonconventional adjuvants.

    Science.gov (United States)

    Sato, Vanessa Sayuri; Jorge, João Atílio; Oliveira, Wanderley Pereira; Souza, Claudia Regina Fernandez; Guimarães, Luis Henrique Souza

    2014-02-28

    Microbial phytases are enzymes with biotechnological interest for the feed industry. In this article, the effect of spray-drying conditions on the stability and activity of extracellular phytase produced by R. microsporus var. microsporus biofilm is described. The phytase was spray-dried in the presence of starch, corn meal (>150 μm), soy bean meal (SB), corn meal (Spray drying of the R. microsporus var. microsporus phytase using different drying adjuvants showed interesting results, being quite feasible with regards their biotechnological applications, especially for poultry diets. PMID:24196167

  3. Recent Advances in Enzymatic Fuel Cells: Experiments and Modeling

    Directory of Open Access Journals (Sweden)

    Ivan Ivanov

    2010-04-01

    Full Text Available Enzymatic fuel cells convert the chemical energy of biofuels into electrical energy. Unlike traditional fuel cell types, which are mainly based on metal catalysts, the enzymatic fuel cells employ enzymes as catalysts. This fuel cell type can be used as an implantable power source for a variety of medical devices used in modern medicine to administer drugs, treat ailments and monitor bodily functions. Some advantages in comparison to conventional fuel cells include a simple fuel cell design and lower cost of the main fuel cell components, however they suffer from severe kinetic limitations mainly due to inefficiency in electron transfer between the enzyme and the electrode surface. In this review article, the major research activities concerned with the enzymatic fuel cells (anode and cathode development, system design, modeling by highlighting the current problems (low cell voltage, low current density, stability will be presented.

  4. Enzymatic hydrolysis of pretreated barley and wheat straw

    DEFF Research Database (Denmark)

    Rosgaard, Lisa

    2007-01-01

    work involved evaluation of 1) possible ways to increase the glucose release from the commercial cellulase product Celluclast by boosting with other enzyme activities to increase the enzymatic hydrolysis, 2) comparing differently pretreated feedstock substrates and 3) evaluating a fed-batch substrate...... feeding strategy to increase the substrate loading in the hydrolysis reaction. The substrate for the enzymatic hydrolysis was primarily steam pretreated wheat and barley straw since these substrates were the primary feedstocks for the Babilafuente Bioethanol process. The initial work showed that there was...... different pretreatment conditions; hot water extraction and acid- or water impregnation followed by steam explosion showed there were slight differences between the effect of pretreatment conditions in relation to the overall yield from enzymatic hydrolysis. The highest glucose concentration was found for...

  5. Metal complexes with 2-acetylpyridine-N(4)-orthochlorophenylthiosemicarbazone: cytotoxicity and effect on the enzymatic activity of thioredoxin reductase and glutathione reductase.

    Science.gov (United States)

    Parrilha, Gabrieli L; Ferraz, Karina S O; Lessa, Josane A; de Oliveira, Kely Navakoski; Rodrigues, Bernardo L; Ramos, Jonas P; Souza-Fagundes, Elaine M; Ott, Ingo; Beraldo, Heloisa

    2014-09-12

    Metal complexes with 2-acetylpyridine-N(4)-orthochlorophenylthiosemicarbazone (H2Ac4oClPh) were assayed for their cytotoxicity against MCF-7 breast adenocarcinoma and HT-29 colon carcinoma cells. The thiosemicarbazone and most of the complexes were highly cytotoxic. H2Ac4oClPh and its gallium(III) and tin(IV) complexes did not show any inhibitory activity against thioredoxin reductase (TrxR) and glutathione reductase (GR). The palladium(II), platinum(II) and bismuth(III) complexes inhibited TrxR at micromolar concentrations but not GR. The antimony(III) and gold(III) complexes strongly inhibited TrxR at submicromolar doses with GR inhibition at higher concentrations. The selectivity of these complexes for TrxR suggests metal binding to a selenol residue in the active site of the enzyme. TrxR inhibition is likely a contributing factor to the mode of action of the gold and antimony derivatives. PMID:25058344

  6. Differences in Enzymatic Properties of the Saccharomyces kudriavzevii and Saccharomyces uvarum Alcohol Acetyltransferases and Their Impact on Aroma-Active Compounds Production

    Science.gov (United States)

    Stribny, Jiri; Querol, Amparo; Pérez-Torrado, Roberto

    2016-01-01

    Higher alcohols and acetate esters belong to the most important yeast secondary metabolites that significantly contribute to the overall flavor and aroma profile of fermented products. In Saccharomyces cerevisiae, esterification of higher alcohols is catalyzed mainly by the alcohol acetyltransferases encoded by genes ATF1 and ATF2. Previous investigation has shown other Saccharomyces species, e.g., S. kudriavzevii and S. uvarum, to vary in aroma-active higher alcohols and acetate esters formation when compared to S. cerevisiae. Here, we aimed to analyze the enzymes encoded by the ATF1 and ATF2 genes from S. kudriavzevii (SkATF1, SkATF2) and S. uvarum (SuATF1, SuATF2). The heterologous expression of the individual ATF1 and ATF2 genes in a host S. cerevisiae resulted in the enhanced production of several higher alcohols and acetate esters. Particularly, an increase of 2-phenylethyl acetate production by the strains that harbored ATF1 and ATF2 genes from S. kudriavzevii and S. uvarum was observed. When grown with individual amino acids as the nitrogen source, the strain that harbored SkATF1 showed particularly high 2-phenylethyl acetate production and the strains with introduced SkATF2 or SuATF2 revealed increased production of isobutyl acetate, isoamyl acetate, and 2-phenylethyl acetate compared to the reference strains with endogenous ATF genes. The alcohol acetyltransferase activities of the individual Atf1 and Atf2 enzymes measured in the cell extracts of the S. cerevisiae atf1 atf2 iah1 triple-null strain were detected for all the measured substrates. This indicated that S. kudriavzevii and S. uvarum Atf enzymes had broad range substrate specificity as S. cerevisiae Atf enzymes. Individual Atf1 enzymes exhibited markedly different kinetic properties since SkAtf1p showed c. twofold higher and SuAtf1p c. threefold higher Km for isoamyl alcohol than ScAtf1p. Together these results indicated that the differences found among the three Saccharomyces species during the

  7. Enzymatic hydrolysis of potato pulp

    OpenAIRE

    Mariusz Lesiecki; Wojciech Białas; Grażyna Lewandowicz

    2012-01-01

    Background. Potato pulp constitutes a complicated system of four types of polysaccharides: cellulose, hemicellulose, pectin and starch. Its composition makes it a potential and attractive raw material for the production of the second generation bioethanol. The aim of this research project was to assess the usefulness of commercial enzymatic preparations for the hydrolysis of potato pulp and to evaluate the effectiveness of hydrolysates obtained in this way as raw materials for ethanol ferment...

  8. Enzymatic fuel cells: Recent progress

    International Nuclear Information System (INIS)

    There is an increasing interest in replacing non-selective metal catalysts, currently used in low temperature fuel cells, with enzymes as catalysts. Specific oxidation of fuel and oxidant by enzymes as catalysts yields enzymatic fuel cells. If the catalysts can be immobilised at otherwise inert anode and cathode materials, this specificity of catalysis obviates the requirement for fuel cell casings and membranes permitting fuel cell configurations amenable to miniaturisation to be adopted. Such configurations have been proposed for application to niche areas of power generation: powering remotely located portable electronic devices, or implanted biomedical devices, for example. We focus in this review on recent efforts to improve electron transfer between the enzymes and electrodes, in the presence or absence of mediators, with most attention on research aimed at implantable or semi-implantable enzymatic fuel cells that harvest the body's own fuel, glucose, coupled to oxygen reduction, to provide power to biomedical devices. This ambitious goal is still at an early stage, with device power output and stability representing major challenges. A comparison of performance of enzymatic fuel cell electrodes and assembled fuel cells is attempted in this review, but is hampered in general by lack of availability of, and conformity to, standardised testing and reporting protocols for electrodes and cells. We therefore highlight reports that focus on this requirement. Ultimately, insight gained from enzymatic fuel cell research will lead to improved biomimetics of enzyme catalysts for fuel cell electrodes. These biomimetics will mimic enzyme catalytic sites and the structural flexibility of the protein assembly surrounding the catalytic site.

  9. Rutin ameliorates glycemic index, lipid profile and enzymatic activities in serum, heart and liver tissues of rats fed with a combination of hypercaloric diet and chronic ethanol consumption.

    Science.gov (United States)

    Chuffa, Luiz Gustavo A; Fioruci-Fontanelli, Beatriz A; Bordon, Juliana G; Pires, Rafaelle B; Braga, Camila P; Seiva, Fábio R F; Fernandes, Ana Angélica H

    2014-06-01

    Alcoholism and obesity are strongly associated with several disorders including heart and liver diseases. This study evaluated the effects of rutin treatment in serum, heart and liver tissues of rats subjected to a combination of hypercaloric diet (HD) and chronic ethanol consumption. Rats were divided into three groups: Control: rats fed a standard diet and drinking water ad libitum; G1: rats fed the HD and receiving a solution of 10% (v/v) ethanol; and G2: rats fed the HD and ethanol solution, followed by injections of 50 mg/kg(-1) rutin as treatment. After 53 days of HD and ethanol exposure, the rutin was administered every three days for nine days. At the end of the experimental period (95 days), biochemical analyses were carried out on sera, cardiac and hepatic tissues. Body weight gain and food consumption were reduced in both the G1 and G2 groups compared to control animals. Rutin effectively reduced the total lipids (TL), triglycerides (TG), total cholesterol (TC), VLDL, LDL-cholesterol and glucose levels, while it increased the HDL-cholesterol in the serum of G2 rats, compared to G1. Although rutin had no effect on total protein, albumin, uric acid and cretinine levels, it was able to restore serum activities of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine kinase (CK) in animals fed HD and receiving ethanol. Glycogen stores were replenished in both hepatic and cardiac tissues after rutin treatment. Moreover, rutin consistently reduced hepatic levels of TG and TC and cardiac AST, ALT and CK activities. Thus, rutin treatment was effective in reducing the risk factors for cardiac and hepatic disease caused by both HD and chronic ethanol consumption. PMID:25204084

  10. Effect of the human therapeutic drug diltiazem on the haematological parameters, histology and selected enzymatic activities of rainbow trout Oncorhynchus mykiss.

    Science.gov (United States)

    Steinbach, Christoph; Burkina, Viktoriia; Schmidt-Posthaus, Heike; Stara, Alzbeta; Kolarova, Jitka; Velisek, Josef; Randak, Tomas; Kroupova, Hana Kocour

    2016-08-01

    Diltiazem is a pharmaceutical belonging to a group of calcium channel blockers (CCB) that is widely used in the treatment of angina pectoris and hypertension. The objective of the present study was to assess the effect of diltiazem on rainbow trout (Oncorhynchus mykiss). Juvenile trout were exposed for 21 and 42 days to three nominal concentrations of diltiazem: 0.03 μg L(-1) (environmentally relevant concentration), 3 μg L(-1), and 30 μg L(-1) (sub-lethal concentrations). The number of mature neutrophilic granulocytes was significantly increased by 450 and 400% in fish exposed to 3 μg L(-1) and 30 μg L(-1) diltiazem compared to the control, respectively. Antioxidant enzyme activity was affected in liver and gills of fish exposed to all tested concentrations of diltiazem but the changes were mostly transient and not concentration dependent. Creatine kinase activity was markedly increased (ranging from 520 to 845%) at all tested diltiazem concentrations at the end of the exposure indicating muscle and/or kidney damage. The highest concentration was associated with histological changes in heart, liver, and kidney. These alterations can be attributed to the effects of diltiazem on the cardiovascular system, similar to those observed in the human body, as well as to its metabolism. At the environmentally relevant concentration, diltiazem was found to induce some alterations in the blood, gills, and liver of fish, indicating its potential for adverse effects on non-target organisms in the aquatic environment. PMID:27208646

  11. Enzymatic transesterification of used frying oils

    Energy Technology Data Exchange (ETDEWEB)

    Kovacs, S.; Hancsok, J. (Univ. of Pannonia, Veszprem (HU)), Email: hancsokj@almos.uni-pannon.hu

    2009-07-01

    The research of converting used frying oils to less harmful products with much higher value was forced by environmental, human biological and economical reasons. One possible pathway of the transformation is the enzymatic transesterification. Through the research work used frying oils (UFO) and sunflower oils (SO) from different origins were first properly pre-treated. Then the previously mentioned feeds and different mixtures of them were transesterified in the presence of Novozym 435 enzyme catalyst under different process conditions. Characteristics of the produced methyl esters were evaluated according to the requirements of EN 14214:2009 standard. We determined that the transesterification of used frying oils is not expediential in the presence of enzyme catalyst because the significant decreasing of catalyst activity. We have found proper UFO and SO mixtures and combination of process conditions (pressure: atmospheric, temperature: 54 +-1 deg C; methanol to triglyceride molar ratio: 4:1; reaction time: 16 hours) resulting in high (>90 %) yield of monoesters. We clearly established that the best results through the enzymatic transesterification were obtained with the improved sunflower oils containing the highest amount (>88 %) of oleic acid and the used frying oils originated from this source. (orig.)

  12. Enzymatic production of human milk oligosaccharides

    DEFF Research Database (Denmark)

    Guo, Yao

    Enzymatic treatment of biomass is an environmentally friendly method to obtain a range of value- added products, such as biofuels, animal feed or food ingredients. The objective of this PhD study was to biocatalytically produce biofunctional food ingredients – human milk oligosaccharides decorated...... with sialic acid from casein glycomacropeptide obtained from dairy side streams. In addition, the biocatalysts employed in this study, i.e. , a sialyltransferase and a sialidase, were subjected to protein engineering to alter the enzyme’s regioselectivity and to improve hydrolase activity, respectively...... glycomacropeptide as a sialyl donor. This is the first study reporting α-2,6-trans-sialidase activity of this enzyme. Using response surface design allowed identification of two differently optimised conditions for PmST-catalysed production of 3'-sialyllactose and 6'-sialyllactose, giving maximum yields of 2.8 m...

  13. Antioxidant Enzymatic Activities and Gene Expression Associated with Heat Tolerance in the Stems and Roots of Two Cucurbit Species (“Cucurbita maxima” and “Cucurbita moschata” and Their Interspecific Inbred Line “Maxchata”

    Directory of Open Access Journals (Sweden)

    Neelam Ara

    2013-12-01

    Full Text Available The elucidation of heat tolerance mechanisms is required to combat the challenges of global warming. This study aimed to determine the antioxidant enzyme responses to heat stress, at the enzymatic activity and gene expression levels, and to investigate the antioxidative alterations associated with heat tolerance in the stems and roots of squashes using three genotypes differing in heat tolerance. Plants of heat-tolerant “C. moschata”, thermolabile “C. maxima” and moderately heat-tolerant interspecific inbred line “Maxchata” genotypes were exposed to moderate (37 °C and severe (42 °C heat shocks. “C. moschata” exhibited comparatively little oxidative damage, with the lowest hydrogen peroxide (H2O2, superoxide (O2− and malondialdehyde (MDA contents in the roots compared to stems, followed by “Maxchata”. The enzyme activities of superoxide dismutase (SOD, ascorbate peroxidase (APX, catalase (CAT and peroxidase (POD were found to be increased with heat stress in tolerant genotypes. The significant inductions of FeSOD, MnSOD, APX2, CAT1 and CAT3 isoforms in tolerant genotypes suggested their participation in heat tolerance. The differential isoform patterns of SOD, APX and CAT between stems and roots also indicated their tissue specificity. Furthermore, despite the sequence similarity of the studied antioxidant genes among “C. maxima” and “Maxchata”, most of these genes were highly induced under heat stress in “Maxchata”, which contributed to its heat tolerance. This phenomenon also indicated the involvement of other unknown genetic and/or epigenetic factors in controlling the expression of these antioxidant genes in squashes, which demands further exploration.

  14. Organic chelants-mediated enhanced lead (Pb) uptake and accumulation is associated with higher activity of enzymatic antioxidants in spinach (Spinacea oleracea L.).

    Science.gov (United States)

    Khan, Imran; Iqbal, Muhammad; Ashraf, Muhammad Yasin; Ashraf, Muhammad Arslan; Ali, Shafaqat

    2016-11-01

    The spinach was tested in the present studies for its phytoextraction potential. Furthermore, the study assessed whether organic chelants could reduce oxidative stress, and thus enhance growth of spinach plants under 2.42 and 4.83mM Pb regimes. Different organic chelates viz. ethylenediamine tetra acetic acid, (EDTA), citric acid (CA), oxalic acid (OA), tartaric acid (TA) and malic acid (MA) were applied separately in addition to control (without chelating agents) under different Pb regimes. The low (2.42mM) Pb regime increased biological yield (kgha(-1)). All the chelates except OA increased biological yield under low Pb regime. In contrast, TA caused less decrease in biomass under high (4.83mM) Pb regime. The chelate-assisted rise in the antioxidant activities substantially contributed to reactive oxygen species (ROS) neutralization. Of the chelates, TA was the most effective in improving Pb uptake and its root to shoot translocation. Overall, the chelate-assisted buildup of Pb in the spinach did not exhibit inhibitory effects on the plant growth possibly due to their potential to decrease Pb-induced oxidative damage. The results elaborated the potential of TA in increasing root to shoot translocation of Pb, biomass, and thus suggested its use for phytoextraction of Pb using spinach in Pb contaminated environments. PMID:27318732

  15. Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS.

    Science.gov (United States)

    Hernández-Ledesma, Blanca; Dávalos, Alberto; Bartolomé, Begoña; Amigo, Lourdes

    2005-02-01

    We have investigated the antioxidant activity of hydrolysates from whey proteins bovine alpha-lactalbumin (alpha-La) and beta-lactoglobulin A (beta-Lg A) by commercial proteases (pepsin, trypsin, chymotrypsin, thermolysin, and Corolase PP). Corolase PP was the most appropriate enzyme to obtain antioxidant hydrolysates from alpha-La and beta-Lg A (ORAC-FL values of 2.315 and 2.151 micromol of Trolox equivalent/mg of protein, respectively). A total of 42 peptide fragments were identified by HPLC-MS/MS in the beta-Lg A hydrolysate by Corolase PP. One of the sequences (Trp-Tyr-Ser-Leu-Ala-Met-Ala-Ala-Ser-Asp-Ile) possessed radical scavenging (ORAC-FL value of 2.621 micromol of Trolox equivalent/micromol of peptide) higher than that of butylated hydroxyanisole (BHA). Our results suggest that whey protein hydrolysates could be suitable as natural ingredients in enhancing antioxidant properties of functional foods and in preventing oxidation reaction in food processing. PMID:15686406

  16. Application of photocatalytic cadmium sulfide nanoparticles to detection of enzymatic activities of glucose oxidase and glutathione reductase using oxidation of 3,3′,5,5′-tetramethylbenzidine

    Energy Technology Data Exchange (ETDEWEB)

    Grinyte, Ruta; Garai-Ibabe, Gaizka; Saa, Laura; Pavlov, Valeri, E-mail: vpavlov@cicbiomagune.es

    2015-06-30

    Highlights: • The light-powered nanosensor fabricated by enzymatic reactions was reported. • The sensor use energy of photons for oxidation of chromogenic enzymatic substrates. • Enzymatic assays for glucose oxidase and glutathione reductase were developed. - Abstract: It was found out that semiconductor CdS nanoparticles (NPs) are able to catalyze photooxidation of the well known chromogenic enzymatic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) by oxygen. The photocatalytical oxidation of TMB does not require hydrogen peroxide and its rate is directly proportional to the quantity of CdS NPs produced in situ through the interaction of Cd{sup 2+} and S{sup 2−} ions in an aqueous medium. This phenomenon was applied to development of colorimetric sensitive assays for glucose oxidase and glutathione reductase based on enzymatic generation of CdS NPs acting as light-powered catalysts. Sensitivity of the developed chromogenic assays was of the same order of magnitude or even better than that of relevant fluorogenic assays. The present approach opens the possibility for the design of simple and sensitive colorimetric assays for a number of enzymes using inexpensive and available TMB as a universal chromogenic compound.

  17. Application of photocatalytic cadmium sulfide nanoparticles to detection of enzymatic activities of glucose oxidase and glutathione reductase using oxidation of 3,3′,5,5′-tetramethylbenzidine

    International Nuclear Information System (INIS)

    Highlights: • The light-powered nanosensor fabricated by enzymatic reactions was reported. • The sensor use energy of photons for oxidation of chromogenic enzymatic substrates. • Enzymatic assays for glucose oxidase and glutathione reductase were developed. - Abstract: It was found out that semiconductor CdS nanoparticles (NPs) are able to catalyze photooxidation of the well known chromogenic enzymatic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) by oxygen. The photocatalytical oxidation of TMB does not require hydrogen peroxide and its rate is directly proportional to the quantity of CdS NPs produced in situ through the interaction of Cd2+ and S2− ions in an aqueous medium. This phenomenon was applied to development of colorimetric sensitive assays for glucose oxidase and glutathione reductase based on enzymatic generation of CdS NPs acting as light-powered catalysts. Sensitivity of the developed chromogenic assays was of the same order of magnitude or even better than that of relevant fluorogenic assays. The present approach opens the possibility for the design of simple and sensitive colorimetric assays for a number of enzymes using inexpensive and available TMB as a universal chromogenic compound

  18. Method to Improve Enzymatic Activity during Beer Brewing%改善啤酒酿造过程酶活力方法的研究

    Institute of Scientific and Technical Information of China (English)

    刘乃侨; 孙丽华

    2011-01-01

    以国外大麦Gairdner为原料,分别用麦芽厂所用的空白水、pH值4.0盐酸溶液、pH值4.0硫酸溶液、100 mg/L钙离子溶液、1μg/L赤霉素溶液使大麦浸渍、发芽来达到降低绿麦芽内部pH值的目的,实验结果表明,虽然硫酸、盐酸2种强酸物质具备降低大麦发芽内部pH值的能力,但其食品安全性差、对设备腐蚀大,而用钙离子和赤霉素溶液培养大麦发芽,虽然在大麦发芽过程中对pH值的变化影响不是很稳定,但均能使大麦发芽结束点即绿麦芽的pH值降低,所以对于后期啤酒的酿造均能够使淀粉酶、蛋白酶等活力近于最适pH值状态,为在啤酒酿造过程中减少外源添加酸的使用量,或为温和型弱酸或酸性中草药等物质的添加提供条件.%The aim of the experiment was to reduce pH value inside green malt during barley germination using Gaird-ner barley as raw material, soaking respectively with fresh water used in the malt factory, pH 4. 0 hydrochloric acid solution, pH 4.0 sulphuric acid solution, 100 mg/L calcium ion solution, and 1 u.g/L gibberellin solution. The results showed that although strong acids of sulphuric acid and hydrochloric acid possesses the capabilities to reduce pH value inside the barley during the germination, yet the food security is poor, and they corrode the facilities greatly, though while using calcium ion and/or gibberellin solution to cultivate barley to germinate had unstable pH changes during the barley germination, but they all could reduce pH value when reaching the end of barley germination I. E. Green malt. Therefore, at the late stage of beer brewing it could reach closely to the most suitable pH value for the activities of amylase and protease, to reduce the adding amount of acid during the beer brewing process, or provide conditions of adding mild weak acids and/or acidic Chinese medical herbs, and other materials.

  19. 酶法水解花生蛋白及产物抗氧化活性的研究%Enzymatic Hydrolysis of Peanut Protein and Its Hydrolysate of Antioxidant Activity

    Institute of Scientific and Technical Information of China (English)

    罗世龙; 张榴萍; 刘锦

    2014-01-01

    Taking extracted peanut protein from peanut meal as raw material, variations of degree of hydrolysis, soluble nitrogen content in trichloroacetic acid, and antioxidant activity at different time were studied by using Alcalase, Protamex, and Papain, respectively. Results showed that degree of hydrolysis of enzymatic hydrolysis product stabilized after increased with extension of hydrolysis time. Soluble nitrogen content in trichloroacetic acid increased first and then decreased. And superoxide anion free radical scavenging capacity increased first and then slightly decreased. There were not directly related between antioxidant properties of different hydrolysis products from different enzyme types and their hydrolysis ability.%本文以花生粕中提取的花生蛋白为原料,分别使用Alcalase、 Protamex和Papain酶解,研究不同酶解时间酶解产物的水解度、三氯乙酸中可溶性氮含量和抗氧化活性的变化规律。结果表明:随着酶解时间的延长, Alcalase、 Protamex和Papain酶解花生蛋白的酶解产物的水解度先逐渐增加后趋于稳定,三氯乙酸中可溶性氮含量先增加后减小,超氧阴离子自由基清除能力先增加后有小幅降低。不同酶的酶解产物的抗氧化性能与其水解能力并不直接相关。

  20. Mechanistic investigation in ultrasound induced enhancement of enzymatic hydrolysis of invasive biomass species.

    Science.gov (United States)

    Borah, Arup Jyoti; Agarwal, Mayank; Poudyal, Manisha; Goyal, Arun; Moholkar, Vijayanand S

    2016-08-01

    This study has assessed four invasive weeds, viz. Saccharum spontaneum (SS), Mikania micrantha (MM), Lantana camara (LC) and Eichhornia crassipes (EC) for enzymatic hydrolysis prior to bioalcohol fermentation. Enzymatic hydrolysis of pretreated biomasses of weeds has been conducted with mechanical agitation and sonication under constant (non-optimum) conditions. Profiles of total reducible sugar release have been fitted to HCH-1 model of enzymatic hydrolysis using Genetic Algorithm. Trends in parameters of this model reveal physical mechanism of ultrasound-induced enhancement of enzymatic hydrolysis. Sonication accelerates hydrolysis kinetics by ∼10-fold. This effect is contributed by several causes, attributed to intense micro-convection generated during sonication: (1) increase in reaction velocity, (2) increase in enzyme-substrate affinity, (3) reduction in product inhibition, and (4) enhancement of enzyme activity due to conformational changes in its secondary structure. Enhancement effect of sonication is revealed to be independent of conditions of enzymatic hydrolysis - whether optimum or non-optimum. PMID:26898160

  1. On the enzymatic hydrolysis of various starches

    Energy Technology Data Exchange (ETDEWEB)

    Tegge, G.; Richter, G.

    1986-10-01

    The behaviour of different commercial starches to amylolytic enzyme preparations and of their hydrolyzates during raffination was investigated. No significant differences in final degree of saccharification of starches from yellow maize, waxy maize, amylo-maize, potatoes and wheat were observed. The lower DE-values of waxy maize hydrolyzates after liquefaction were completely compensated during final saccharification phase. Determinations of viscosity after liquefaction and saccharification always showed highest viscosity in raw hydrolyzates these differences in viscosity were no more observed. Addition of pentosanase during saccharification period did not affect viscosity and filtration of the hydrolyzates. Glucoamylase with increased pentosanase activity affected filtration of wheat starch hydrolyzates positively; viscosity kept unchanged. Development of enzymatic liquefaction of the individual starches was studied by means of a Brabender Viscograph. By this informative differences between potatoe and waxy maize starches on one side and maize and wheat starches on the other side were observed.

  2. Enzymatic activity of mycorrhizal fungi. II

    Directory of Open Access Journals (Sweden)

    R. Pachlewski

    2014-08-01

    Full Text Available An attempt bas been made to evaluate the participation of mycorrhizal fungi in the process of degradation of some aromatic compounds. 24 strains of ectomycorrhizal fungi, l ectendomycorrhizal stram and 11 nonmycorrhizal strains which decomposed wood were investigated. The observations were aimed at showing the synthesis by these fungi of laccase, peroxidase and tyrosinase. None of these fungi synthetized peroxidase.

  3. Ultrasound-enhanced enzymatic hydrolysis of poly(ethylene terephthalate).

    Science.gov (United States)

    Pellis, Alessandro; Gamerith, Caroline; Ghazaryan, Gagik; Ortner, Andreas; Herrero Acero, Enrique; Guebitz, Georg M

    2016-10-01

    The application of ultrasound was found to enhance enzymatic hydrolysis of poly(ethylene terephthalate) (PET). After a short activation phase up to 6.6times increase in the amount of released products was found. PET powder with lower crystallinity of 8% was hydrolyzed faster when compared to PET with 28% crystallinity. Ultrasound activation was found to be around three times more effective on powders vs. films most likely due to a larger surface area accessible to the enzyme. PMID:27481467

  4. Enzymatic synthesis of magnetic nanoparticles.

    Science.gov (United States)

    Kolhatkar, Arati G; Dannongoda, Chamath; Kourentzi, Katerina; Jamison, Andrew C; Nekrashevich, Ivan; Kar, Archana; Cacao, Eliedonna; Strych, Ulrich; Rusakova, Irene; Martirosyan, Karen S; Litvinov, Dmitri; Lee, T Randall; Willson, Richard C

    2015-01-01

    We report the first in vitro enzymatic synthesis of paramagnetic and antiferromagnetic nanoparticles toward magnetic ELISA reporting. With our procedure, alkaline phosphatase catalyzes the dephosphorylation of l-ascorbic-2-phosphate, which then serves as a reducing agent for salts of iron, gadolinium, and holmium, forming magnetic precipitates of Fe45±14Gd5±2O50±15 and Fe42±4Ho6±4O52±5. The nanoparticles were found to be paramagnetic at 300 K and antiferromagnetic under 25 K. Although weakly magnetic at 300 K, the room-temperature magnetization of the nanoparticles found here is considerably greater than that of analogous chemically-synthesized LnxFeyOz (Ln = Gd, Ho) samples reported previously. At 5 K, the nanoparticles showed a significantly higher saturation magnetization of 45 and 30 emu/g for Fe45±14Gd5±2O50±15 and Fe42±4Ho6±4O52±5, respectively. Our approach of enzymatically synthesizing magnetic labels reduces the cost and avoids diffusional mass-transfer limitations associated with pre-synthesized magnetic reporter particles, while retaining the advantages of magnetic sensing. PMID:25854425

  5. Enzymatic Synthesis of Magnetic Nanoparticles

    Directory of Open Access Journals (Sweden)

    Arati G. Kolhatkar

    2015-04-01

    Full Text Available We report the first in vitro enzymatic synthesis of paramagnetic and antiferromagnetic nanoparticles toward magnetic ELISA reporting. With our procedure, alkaline phosphatase catalyzes the dephosphorylation of l-ascorbic-2-phosphate, which then serves as a reducing agent for salts of iron, gadolinium, and holmium, forming magnetic precipitates of Fe45±14Gd5±2O50±15 and Fe42±4Ho6±4O52±5. The nanoparticles were found to be paramagnetic at 300 K and antiferromagnetic under 25 K. Although weakly magnetic at 300 K, the room-temperature magnetization of the nanoparticles found here is considerably greater than that of analogous chemically-synthesized LnxFeyOz (Ln = Gd, Ho samples reported previously. At 5 K, the nanoparticles showed a significantly higher saturation magnetization of 45 and 30 emu/g for Fe45±14Gd5±2O50±15 and Fe42±4Ho6±4O52±5, respectively. Our approach of enzymatically synthesizing magnetic labels reduces the cost and avoids diffusional mass-transfer limitations associated with pre-synthesized magnetic reporter particles, while retaining the advantages of magnetic sensing.

  6. Enzymatic Modification of Polyethersulfone Membranes

    Directory of Open Access Journals (Sweden)

    Karin Schroën

    2012-11-01

    Full Text Available Enzymatic modification of polyethersulfone (PES membranes has been found not only feasible, but also an environmentally attractive way to vary surface properties systematically. In this paper, we summarize the effect of modification layers on protein adsorption and bacterial adhesion on PES membranes and surfaces. The enzyme laccase was used to covalently bind (polyphenolic acids to the membrane, and compared to other membrane modification methods, this method is very mild and did not influence the mechanical strength negatively. Depending on the conditions used during modification, the modification layers were capable of influencing interactions with typical fouling species, such as protein, and to influence attachment of microorganisms. We also show that the modification method can be successfully applied to hollow fiber membranes; and depending on the pore size of the base membrane, proteins were partially rejected by the membrane. In conclusion, we have shown that enzymatic membrane modification is a versatile and economically attractive method that can be used to influence various interactions that normally lead to surface contamination, pore blocking, and considerable flux loss in membranes.

  7. SOIL QUALITY ASSESSMENT BASED ON CHEMICAL, ENZYMATIC AND BACTERIOLOGICAL ANALYSIS

    Directory of Open Access Journals (Sweden)

    Sofia-Paulina BALAURE

    2012-01-01

    Full Text Available This study highlights the problem of soil pollution as the result of human activities. Soil pollutans may be either chemicals or biological in nature. microbial enzymatic activities are often proposed as indicators of environmental stress. The soil samples were submitted by chemical, microbiological and enzymatic analyses. Chemical analyses were been made for determinating the heavy metals. Heavy metals from the forest soil were represented by Cu, Zn, Mn, Ni, Pb, Cd and Cr. To evaluate the concentration in heavy metals from the filtrate, we used a acetylene-nitrous oxide flame atomic absorption spectrophotometry. Potential dehydrogenase activity, the only indicator of the possible sources of pollution, excluded the presence of either chemical or biological pollution. The number of bacteria involved in the biogeochemical cycle of nitrogen in the analyzed soil indicated a high efficiency regarding the mineralization of the organic residues of plant and animal origin.

  8. Ultrasound assisted enzymatic depolymerization of aqueous guar gum solution.

    Science.gov (United States)

    Prajapat, Amrutlal L; Subhedar, Preeti B; Gogate, Parag R

    2016-03-01

    The present work investigates the effectiveness of application of low intensity ultrasonic irradiation for the intensification of enzymatic depolymerization of aqueous guar gum solution. The extent of depolymerization of guar gum has been analyzed in terms of intrinsic viscosity reduction. The effect of ultrasonic irradiation on the kinetic and thermodynamic parameters related to the enzyme activity as well as the intrinsic viscosity reduction of guar gum using enzymatic approach has been evaluated. The kinetic rate constant has been found to increase with an increase in the temperature and cellulase loading. It has been observed that application of ultrasound not only enhances the extent of depolymerization but also reduces the time of depolymerization as compared to conventional enzymatic degradation technique. In the presence of cellulase enzyme, the maximum extent of depolymerization of guar gum has been observed at 60 W of ultrasonic rated power and ultrasonic treatment time of 30 min. The effect of ultrasound on the kinetic and thermodynamic parameters as well as the molecular structure of cellulase enzyme was evaluated with the help of the chemical reaction kinetics model and fluorescence spectroscopy. Application of ultrasound resulted in a reduction in the thermodynamic parameters of activation energy (Ea), enthalpy (ΔH), entropy (ΔS) and free energy (ΔG) by 47%, 50%, 65% and 1.97%, respectively. The changes in the chemical structure of guar gum treated using ultrasound assisted enzymatic approach in comparison to the native guar gum were also characterized by FTIR. The results revealed that enzymatic depolymerization of guar gum resulted in a polysaccharide with low degree of polymerization, viscosity and consistency index without any change in the core chemical structure which could make it useful for incorporation in food products. PMID:26584988

  9. Role of enzymatic and non enzymatic antioxidant in ameliorating salinity induced damage in nostoc muscorum

    International Nuclear Information System (INIS)

    Presence of high salt concentration in the growth medium adversely affected the plant growth and productivity by altering its metabolic activities. Experiments were conducted on cyanobacteriaum Nostoc muscorum grown in nitrogen free medium supplemented with 250 mM NaCl to evaluate the salt stress induced changes in growth, antioxidants and lipid composition. Salt stress significantly reduced the growth and physio-biochemical attributes. Salt stress increased malonaldehyde content thereby causing alterations in the lipid fraction. Significant reduction in polyunsaturated fatty acids including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI) and phosphatidylserine (PS) was observed. Where as diacylglycerol, sterol ester and non-esterified fatty acids were increased. Activities of antioxidant enzymes and contents of non-enzymatic antioxidants including glutathione enhanced due to salt stress. An increase in accumulation of proline was also observed. Hence increased activity of antioxidants and altered fatty acid composition was observed in salt stressed Nostoc muscorum. (author)

  10. Synthesis of monoacylglycerols by enzymatic methods

    Directory of Open Access Journals (Sweden)

    Bradić Milena R.

    2010-01-01

    Full Text Available Monoacylglycerols are non-ionic surfactants widely used in the food industry. They are also important in cosmetic and pharmaceutical industries as drug carriers and for the consistency improvements in creams and lotions. Current process for their production is based on the glycerolysis of natural fats and oils in the presence of inorganic catalysts at temperatures higher than 220 oC. The major drawbacks of this process include high-energy consumption, low yield, and poor product quality. The use of lipases for the monoacylglycerols production offers environmental advantages and a reduction in energy consumption. Besides, the same surfactants prepared by the enzymatic synthesis may be labeled as “natural”. Recent progress in the application of highly-stable lipases in the organic solvents offers the possibility of employing various methods to the enzyme-catalyzed synthesis of monoacylglycerols, such as selective hydrolysis of fats and oils using 1,3-regiospecific lipases, the esterification of glycerol with fatty acids and the glycerolysis of fats or oils. In this review, different reaction systems such as aqueous-organic two-phase systems, microemulsions and reverse micelles systems, anhydrous organic solvents, solvent-free systems with free or immobilized lipases, as well as the use of two-phase membrane reactor systems are presented. We discuss some of the key factors, such as the control of water content, removing of the products from reaction system, and the effects of solvent on the lipase activity and selectivity, that must be addressed in order to obtain an efficient reaction system with high yields of monoacylglycerols. Engineering of the enzymatic monoacylglycerols synthesis processes requires also optimization of other factors as: molar ratio of substrates, temperature, type of lipase immobilization and supports (if any, reactor design and operating regime.

  11. Characterizing Enzymatic Deposition for Microelectrode Neurotransmitter Detection

    Energy Technology Data Exchange (ETDEWEB)

    Hosein, W. K. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Yorita, A. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Tolosa, V. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2016-08-12

    The enzyme immobilization process, one step in creating an enzymatic biosensor, was characterized and analyzed as a function of its physical properties. The neural glutamic biosensor is a flexible device, effectively minimizing trauma to the area of implantation. The Multielectrode Array (MEA) is composed primarily of a proprietary polymer which has been successfully implanted into human subjects in recent years. This polymer allows the device the pliability that other devices normally lack, though this poses some challenges to implantation. The electrodes are made of Platinum (Pt), and can range in number from eight to thirty two electrodes per device. These electrodes are electroplated with a semipermeable polymer layer to improve selectivity of the electrode to the neurotransmitter of interest, in this case glutamate. A signal is created from the interaction of glutamate in the brain with the glutamate oxidase (GluOx) which is immobilized on the surface of the electrode by using crosslinking chemistry in conjunction with glutaraldehyde and Bovine Serum Albumin (BSA). The glutamate is oxidized by glutamate oxidase, producing α-ketoglutarate and hydrogen peroxide (H2O2) as a by-product. The production of H2O2 is crucial for detection of the presence of the glutamate within the enzymatic coating, as it diffuses through the enzyme layer and oxidizes at the surface of the electrode. This oxidation is detectable by measurable change in the current using amperometry. Hence, the MEA allows for in vivo monitoring of neurotransmitter activity in real time. The sensitivity of the sensor to these neurotransmitters is dependent on the thickness of the layer, which is investigated in these experiments in order to optimize the efficacy of the device to detecting the substrate, once implanted.

  12. ENZYMATIC HYDROLYSIS OF AGRICULTURAL LIGNOCELLULOSIC BIOMASS

    Directory of Open Access Journals (Sweden)

    S. STRAVA

    2009-05-01

    Full Text Available The yield, productivity and cost for the enzymatic hydrolysis of cellulose to glucoseare crucial for the production of second generation ethanol. In the first study wehave evaluated the activity of several commercial cellulolytic enzymes and a crudeextract of a local strain of Trichoderma viride. The load used was 15 U ofcellulase/gram cellulose and 90 U of cellobiase/gram cellulose. The hydrolysis wascarried out at 50oC and pH 4,8 for 96 hours. The best cellulose hydrolysis yield of58% was obtained with the cocktail formed of crude cellulases from T. virideCMIT3.5 combined with Novozyme 188. This cocktail was used in the second study,when alkaline-steam pretreated wheat straw and corn stover where hydrolyzed at pH4,8 for 96 hours. The temperature was set at 50oC and 40oC. The hydrolysis at lowertemperature was tested for a future experiment of simultaneous hydrolysis andfermentation. An enzymatic assay using glucose-6-phosphate dehydrogenase wasused to determine exclusively glucose, instead of wide-range sugar DNS assay.Reporting to 100 grams of wet pretreated biomass, the following results wereobtained: 14.4 g% glucose for corn stover at 50oC and 13,0 g% at 40oC; 13,1 g%glucose for wheat straw at 50oC and 10.3 g% at 40oC. Considering that wheat strawcontain 36.6% glucose-based carbohydrates, the hydrolysis yields are between39.3% and 28.1%. Further studies, concerning the optimal parameters for cellulasecocktail will be made.

  13. Enzymatic acylglycerol synthesis in membrane reactor systems.

    NARCIS (Netherlands)

    Padt, van der A.

    1993-01-01

    Up till twenty years ago, only chemical modifications of agricultural oils for novel uses were studied. Because of the instability of various fatty acids, enzymatic biomodifications can have advantages above the chemical route. Nowadays, enzymatic catalysis can be used for the modification of oils a

  14. Non-enzymatic amperometric glucose biosensor from zinc oxide nanoparticles decorated multi-walled carbon nanotubes.

    Science.gov (United States)

    Baby, Tessy Theres; Ramaprabhu, S

    2011-06-01

    The present work describes the development of novel ZnO dispersed multi-walled carbon nanotubes (MWNT) based non-enzymatic glucose biosensor with 1 M NaOH solution as the supporting electrolyte. For a comparison, the same material has been used for the fabrication of enzymatic biosensor and studied its electrochemical activity with phosphate buffer solution as the electrolyte. MWNT have been synthesized by catalytic chemical vapor decomposition (CCVD) and a simple sol-gel method was used for decorating crystalline ZnO nanoparticles on MWNT. Cyclic voltammetry and chronoamperometry were used to study and optimize the electrochemical performance of the resulting enzymatic and non-enzymatic ZnO/MWNT biosensors. The non enzymatic Nafion/ZnO/MWNT/GC electrode shows linearity in the range 700 nM to 31 mM with the detection limit of 500 nM. Similarly enzymatic biosensor fabricated using Nafion/GOD/ZnO/MWNT on glassy carbon electrode (GCE) shows a linearity from 1 microM to 22 mM. This excellent performance of non enzymatic Nafion/ZnO/MWNT/GC is due to high surface area, good electron transfer rate of ZnO/MWNT and the high electrochemical catalytic activity of ZnO in NaOH solution. PMID:21770093

  15. Enzymatic hydrolysis of steryl glycosides for their analysis in foods.

    Science.gov (United States)

    Münger, Linda H; Nyström, Laura

    2014-11-15

    Steryl glycosides (SG) contribute significantly to the total intake of phytosterols. The standard analytical procedure involving acid hydrolysis fails to reflect the correct sterol profile of SG due to isomerization of some of the labile sterols. Therefore, various glycosylases were evaluated for their ability to hydrolyse SG under milder conditions. Using a pure SG mixture in aqueous solution, the highest glycolytic activity, as demonstrated by the decrease in SG and increase in free sterols was achieved using inulinase preparations (decrease of >95%). High glycolytic activity was also demonstrated using hemicellulase (63%). The applicability of enzymatic hydrolysis using inulinase preparations was further verified on SG extracted from foods. For example in potato peel Δ(5)-avenasteryl glucoside, a labile SG, was well preserved and contributed 26.9% of the total SG. Therefore, enzymatic hydrolysis is suitable for replacing acid hydrolysis of SG in food lipid extracts to accurately determine the sterol profile of SG. PMID:24912717

  16. Enzymatic Catalysis of Proton Transfer and Decarboxylation Reactions

    OpenAIRE

    Richard, John P.

    2011-01-01

    Deprotonation of carbon and decarboxylation at enzyme active sites proceed through the same carbanion intermediates as for the uncatalyzed reactions in water. The mechanism for the enzymatic reactions can be studied at the same level of detail as for nonenzymatic reactions, using the mechanistic tools developed by physical organic chemists. Triosephosphate isomerase (TIM) catalyzed interconversion of D-glyceraldehyde 3-phosphate and dihydroxyacetone phosphate is being studied as a prototype f...

  17. Pre and Postharvest Enzymatic Activity in Gulupa (Passiflora edulis Sims Fruits from the Colombian Lower Montane Rain Forest / Actividad Enzimática Precosecha y Poscosecha en Frutos de Gulupa (Passiflora edulis Sims, en Condiciones del Bosque Húmedo Mo

    Directory of Open Access Journals (Sweden)

    Germán Franco

    2014-03-01

    Full Text Available “High-Andean fruits” are deemed important because oftheir potential for domestic consumption and exportation. Among them, gulupa (Passiflora edulis Sims is an exotic fruit of good acceptance in European markets. However, the technological support associated with the crop is incipient and its short shelf life leads to rapid deterioration of the product. This fact makes it necessary to investigate the physical, physiological and biochemical processes that characterize fruit ripening, in order to take actions to ensure that it arrives in its best possible condition to the consumer. In this context, the current study aimed at identifying enzymatic activity in gulupa fruits during pre and postharvest. Plant materialfrom the Colombian Gene Bank (administered by Corpoica was used. Fruits of known age were periodically harvested to determine the activity of the enzymes α-amylase, polygalacturonase (PG, pectinmethylesterase (PME and polyphenol oxidase (PPO through destructive samplings. It was found that α-amylase and PG are linked to the increase of soluble solids, which favors the sweet taste of the fruit. In turn, the low activity of PPO enables agroindustrial processing without severe fruit browning. / Los “frutales alto-andinos”, se consideran importantes por su potencial de consumo nacional y exportación. Entre ellos está la gulupa (Passiflora edulis Sims, reconocida como un frutal exótico de buena aceptación en mercados europeos. Sin embargo, el respaldo tecnológico asociado al cultivo, es incipiente y su corta vida poscosecha conduce al rápido deterioro del producto. Esto hace necesario plantear estudios de los procesos físicos, fisiológicos y bioquímicos que caracterizan la maduración, con el fin de procurar que el fruto llegue en las mejores condiciones de calidad a los consumidores. El estudio tuvo como objetivo conocer la actividad enzimática en los frutos de gulupa en precosecha y en poscosecha, con el

  18. Enzymatic Extraction and Antibacterial Activity of Aucubin from Eucommia ulmoides Leaves%杜仲叶桃叶珊瑚苷的酶法提取及其抑菌活性

    Institute of Scientific and Technical Information of China (English)

    郑杰; 刘端; 赵肃清; 苏娟; 颜秋萍; 陈磊; 肖奕; 张春梅

    2012-01-01

    Objective: To investigate the technology of Aucubin in Eucommia ulmoides leaves extracted by enzymatic method and its antibacterial activity. Methods: Aucubin in Eucommia ulmoides leaves was extracted by cellulase method. The extraction technology was optimized using the content of Aucubin as index. The antibacterial activity of Aucubin was determined. Results:The results showed that the optimum technology was as follows; The solid-liquid ratio was 1:12, extracted for 50 min by 0.4% enzyme at 50 ℃ in pH 6.0. The extraction rate of Aucubin was as high as 17. 892 mg/g. The Aucubin extracted could obviously inhibit the growth of Escherichia coli and Staphylococcus aureus, the MIC of Aucubin against Staphylococcus aureus and Escherichia coli were 4.832 mg/mL and 9.664 mg/mL respectively. However, Aucubin presented weak inhibitory effect on Streptococcus pneumonia and MG-hemolytic streptococcus, the MIC of Aucubin against them were all 28.946 mg/mL. Conclusion:The extraction technology obtained in this experiment is reasonable and feasible with high extraction rate, and the Aucubin has some antibacterial activity.%目的:研究杜仲叶桃叶珊瑚苷的酶法提取工艺及其抑菌活性.方法:采用纤维素酶法,以桃叶珊瑚苷提取率为指标,优选最佳提取条件,并对其抑菌效果进行研究.结果:杜仲叶桃叶珊瑚苷的最佳提取工艺为:料液比1:12,酶解pH6.0,酶用量0.4%,酶解温度50℃,酶解时间50 min,溶出量可达17.892 mg/g.桃叶珊瑚苷提取液对大肠杆菌和金黄色葡萄球菌的抑菌作用较强,对金黄色葡萄球菌的MIC为4.832 mg/mL,对大肠杆菌的MIC为9.664 mg/mL,但对肺炎链球菌和MG溶血性链球菌的抑菌作用不明显,MIC均为28.946 mg/mL.结论:该提取工艺合理可行,提取率高,提取得到的桃叶珊瑚苷具有一定的抑菌作用.

  19. Production of Fish Hydrolysates Protein from Waste of Fish Carp (Cyprinus Carpio) By Enzymatic Hydrolysis

    OpenAIRE

    Dede Saputra; Tati Nurhayati

    2016-01-01

    Fish Protein Hydrolysates (FPH) is the mixed products of polypeptide, dipeptides, and amino acid. It can be produced from materials that contained of protein by acid reaction, base reaction or enzymatic hydrolysis. The objectives of this study were to study the production of FPH from fish carp meat at post rigor phase and viscera by enzymatic hydrolysis, to determine the specific activity of papain enzyme, and to determine the solubility of FPH. Capacity of fish hydrolyzing can be...

  20. Physiological and enzymatic analyses of pineapple subjected to ionizing radiation

    International Nuclear Information System (INIS)

    The physiological and enzymatic post-harvest characteristics of the pineapple cultivar Smooth Cayenne were evaluated after the fruits were gamma-irradiated with doses of 100 and 150 Gy and the fruits were stored for 10, 20 and 30 days at 12 deg C (±1) and relative humidity of 85% (±5). Physiological and enzymatic analyses were made for each storage period to evaluate the alterations resulting from the application of ionizing radiation. Control specimens showed higher values of soluble pectins, total pectins, reducing sugars, sucrose and total sugars and lower values of polyphenyloxidase and polygalacturonase enzyme activities. All the analyses indicated that storage time is a significantly influencing factor. The 100 Gy dosage and 20-day storage period presented the best results from the standpoint of maturation and conservation of the fruits quality. (author)

  1. IMPORTANCE OF ENZYMATIC BIOTRANSFORMATION IN IMMUNOTOXICOLOGY

    Science.gov (United States)

    Many immunotoxic compounds, such as benzene and other organic solvents, pesticides, mycotoxins and polycyclic aromatic hydrocarbons, can alter immune function only after undergoing enzyme-mediated reactions within various tissues. In the review that follows, the role of enzymatic...

  2. Enzymatic hydrolysis of plant extracts containing inulin

    Energy Technology Data Exchange (ETDEWEB)

    Guiraud, J.P.; Galzy, P.

    1981-10-01

    Inulin-rich extracts of chicory and Jerusalem artichoke are a good potential source of fructose. Total enzymatic hydrolysis of these extracts can be effected by yeast inulinases (EC 3.2.1.7). Chemical prehydrolysis is unfavourable. Enzymatic hydrolysis has advantages over chemical hydrolysis: it does not produce a dark-coloured fraction or secondary substances. It is possible to envisage the preparation of high fructose syrups using this process. (Refs. 42).

  3. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    OpenAIRE

    Neeharika, T. S.V.R.; Lokesh, P.; Prasanna Rani, K. N.; Prathap Kumar, T.; Prasad, R. B.N.

    2015-01-01

    Ricinoleic acid is an unsaturated hydroxy fatty acid that naturally occurs in castor oil in proportions of up to 85–90%. Ricinoleic acid is a potential raw material and finds several applications in coatings, lubricant formulations and pharmaceutical areas. Enzymatic hydrolysis of castor oil is preferred over conventional hydrolysis for the preparation of ricinoleic acid to avoid estolide formation. A kinetics analysis of the enzymatic hydrolysis of Methyl Ricinoleate in the presence of Candi...

  4. Enzymatic modification and characterization of xanthan

    OpenAIRE

    Kool, M.M.

    2014-01-01

    In this thesis an enzymatic approach for the modification and characterization of xanthans was introduced. Complete backbone degradation of xanthan by cellulases was obtained independent on the molar composition of a xanthan sample. It was shown that only xanthan segments that occurred in a disordered xanthan conformation were susceptible to enzymatic backbone degradation. HILIC-ELSD-MS analysis revealed the presence of six different xanthan repeating units (RUs). All RUs consisted of the sam...

  5. Preparation and Enzymatic Degradation of Porous Crosslinked Polylactides of Biomass Origin

    Directory of Open Access Journals (Sweden)

    Yuya Kido

    2014-06-01

    Full Text Available To understand the enzymatic degradation behavior of crosslinked polylactide (PLA, the preparation and enzymatic degradation of both thermoplastic (linear and crosslinked PLAs that have pore structures with different dimensions were carried out. The porous structures of the linear PLA samples were of micro and nanoporous nature, and prepared by batch foaming with supercritical CO2 and compared with the porous structures of crosslinked PLA (Lait-X created by the salt leaching method. The surface and cross-sectional morphologies of the porous structures were investigated by using scanning electron microscopy. The morphological analysis of porous Lait-X showed a rapid loss of physical features within 120 h of exposure to proteinase-K enzymatic degradation at 37 °C. Due to the higher affinity for water, enhanced enzymatic activity as compared to the linear PLA porous structures in the micro and nanoporous range was observed.

  6. Enzymatic technologies for remediation of hydrophobic organic pollutants in soil.

    Science.gov (United States)

    Eibes, G; Arca-Ramos, A; Feijoo, G; Lema, J M; Moreira, M T

    2015-11-01

    Worldwide there are numerous contaminated sites as a result of the widespread production and use of chemicals in industrial and military activities as well as poor schemes of waste disposal and accidental spillages. The implementation of strategies for decontamination and restoration of polluted sites has become a priority, being bioremediation with biological agents a promising alternative. Enzyme-based technologies offer several advantages over the use of microbial cells, provided that the biocatalyst meets specific requirements: efficiency to remove the target pollutant/s, non-dependency on expensive coenzymes or cofactors, enzyme stability, and an affordable production system. In this mini-review, the direct application of enzymes for in situ soil bioremediation is explored, and also novel ex situ enzymatic technologies are presented. This new perspective provides a valuable insight into the different enzymatic alternatives for decontamination of soils. Examples of recent applications are reported, including pilot-scale treatments and patented technologies, and the principles of operation and the main requirements associated are described. Furthermore, the main challenges regarding the applicability of enzymatic technologies for remediation of hydrophobic organic pollutants from soil are discussed. PMID:26293336

  7. Enzymatic hydrolysis of protein:mechanism and kinetic model

    Institute of Scientific and Technical Information of China (English)

    Qi Wei; He Zhimin

    2006-01-01

    The bioreaction mechanism and kinetic behavior of protein enzymatic hydrolysis for preparing active peptides were investigated to model and characterize the enzymatic hydrolysis curves.Taking into account single-substrate hydrolysis,enzyme inactivation and substrate or product inhibition,the reaction mechanism could be deduced from a series of experimental results carried out in a stirred tank reactor at different substrate concentrations,enzyme concentrations and temperatures based on M-M equation.An exponential equation dh/dt = aexp(-bh) was also established,where parameters a and b have different expressions according to different reaction mechanisms,and different values for different reaction systems.For BSA-trypsin model system,the regressive results agree with the experimental data,i.e.the average relative error was only 4.73%,and the reaction constants were determined as Km = 0.0748 g/L,Ks = 7.961 g/L,kd = 9.358/min,k2 =38.439/min,Ea= 64.826 kJ/mol,Ed= 80.031 kJ/mol in accordance with the proposed kinetic mode.The whole set of exponential kinetic equations can be used to model the bioreaction process of protein enzymatic hydrolysis,to calculate the thermodynamic and kinetic constants,and to optimize the operating parameters for bioreactor design.

  8. Recent Research Trends on the Enzymatic Synthesis of Structured Lipids.

    Science.gov (United States)

    Kim, Byung Hee; Akoh, Casimir C

    2015-08-01

    Structured lipids (SLs) are lipids that have been chemically or enzymatically modified from their natural biosynthetic form. Because SLs are made to possess desired nutritional, physicochemical, or textural properties for various applications in the food industry, many research activities have been aimed at their commercialization. The production of SLs by enzymatic procedures has a great potential in the future market because of the specificity of lipases and phospholipases used as the biocatalysts. The aim of this review is to provide concise information on the recent research trends on the enzymatic synthesis of SLs of commercial interest, such as medium- and long-chain triacylglycerols, human milk fat substitutes, cocoa butter equivalents, trans-free or low-trans plastic fats (such as margarines and shortenings), low-calorie fats/oils, health-beneficial fatty acid-rich fats/oils, mono- or diacylglycerols, and structurally modified phospholipids. This limited review covers 108 research articles published between 2010 and 2014 which were searched in Web of Science. PMID:26189491

  9. 鳄鱼血蛋白酶解产物抗氧化特性和血管紧张素转化酶抑制活性研究%Study on the antioxidant properties and angiotensin-converting enzyme inhibitory activity of crocodile blood protein enzymatic hydrolysate

    Institute of Scientific and Technical Information of China (English)

    黄和平; 陈孙福; 罗永康

    2014-01-01

    研究了鳄鱼血蛋白酶解产物的抗氧化特性和对血管紧张素转化酶( ACE)的抑制活性。利用木瓜蛋白酶酶解鳄鱼血浆蛋白和血球蛋白,用分光光度法测定了酶解产物的抗氧化能力和用高效液相色谱( HPLC)测定其ACE的抑制率。结果显示:鳄鱼血浆和血球蛋白酶解产物的亚铁离子螯合能力差异性不显著( P>0.05);在0~5 mg/mL的浓度范围内,血球蛋白酶解产物清除ABTS自由基的能力大于血浆蛋白酶解产物,且在浓度为1 mg/mL时,两者清除ABTS自由基的能力差异性极显著( P<0.01);血浆蛋白酶解产物清除DPPH自由基的能力在0~5 mg/mL的浓度范围内随着蛋白浓度的增加而升高,血球蛋白酶解产物在蛋白浓度为4 mg/mL处达到最大清除率,之后下降;在0~20 mg/mL的浓度范围内,两种酶解产物的还原力随着蛋白浓度的提高显著升高,但两者还原力的差异性不显著( P>0.05);鳄鱼血浆和血球蛋白酶解产物对ACE具有良好的抑制力,其最大抑制率可分别达到75.56%和86.42%。研究表明,鳄鱼血蛋白酶解产物在体外具有抗氧化和抑制ACE的活性。%The antioxidant properties and angiotensin-converting enzyme ( ACE) inhibitory activity of enzymatic hydrolysate from crocodile blood protein were analyzed. The crocodile plasma and blood cell protein were hydrolyzed by papain, and then antioxidant properties and ACE inhibition rate of enzymatic hydrolysate were measured by spectrophotometer and HPLC. Results showed that there were no statistical significance ( P>0.05) between the enzymatic hydrolysate of crocodile plasma and blood cell protein; the ABTS radical-scavenging ability of enzymatic hydrolysate from blood cell protein was higher than that from crocodile plasma protein from 0 to 5 mg/mL of protein concentration, and there were statistical significance at P0.05) between them ; crocodile plasma

  10. Enhancing fermentable sugar yield from cassava pulp for bioethanol production: microwave-coupled enzymatic hydrolysis approach.

    Science.gov (United States)

    Sudha, A; Sivakumar, V; Sangeetha, V; Devi, K S Priyenka

    2015-08-01

    Cassava pulp, a potential biological feedstock for ethanol production has been subjected to microwave-assisted alkali pretreatment and microwave-coupled enzymatic hydrolysis. Microwave pretreatment may be a good alternative as it can reduce the pretreatment time and improve the enzymatic activity during hydrolysis. Liquid to solid ratio for the pretreatment of cassava pulp was found to be 20:1. Cassava pulp was pretreated at various NaOH concentration, microwave temperature and gave maximum yield of reducing sugar with 1.5% NaOH at 90 °C in 30 min than conventional alkali pretreatment after enzymatic hydrolysis. The subsequent enzymatic saccharification of pretreated cassava pulp using α amylase dosage of 400 IU at microwave temperature of 90 °C resulted in highest reducing sugar yield of 723 mg/g pulp. Microwave-assisted alkali pretreatment improved the enzymatic saccharification of cassava pulp by increasing its accessibility to hydrolytic enzymes. Microwave-assisted alkali pretreatment and microwave-coupled enzymatic hydrolysis are found to be efficient for improving the yield of reducing sugar. PMID:25832789

  11. Operation and Control of Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Huusom, Jakob Kjøbsted; Nordblad, Mathias;

    This work explores the control of biodiesel production via an enzymatic catalyst. The process involves the transesterification of oils/fats with an alcohol (usually methanol or ethanol), using enzymatic catalysts to generate mono-alkyl esters (the basis of biodiesel) and glycerol as by......-product. Current literature indicates that enzymatic processing of oils and fats to produce biodiesel is technically feasible and developments in immobilization technology indicate that enzyme catalysts can become cost effective compared to chemical processing. However, with very few exceptions, enzyme technology...... is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process designs, which do not use the full potential of the catalysts in a cost-efficient way. Furthermore is it unclear what process variables need to be monitored and controlled to ensure optimal economics...

  12. Enzymatic biodiesel production: Technical and economical considerations

    DEFF Research Database (Denmark)

    Munk Nielsen, Per; Brask, Jesper; Fjerbæk, Lene

    2008-01-01

    It is well documented in the literature that enzymatic processing of oils and fats for biodiesel is technically feasible. However, with very few exceptions, enzyme technology is not currently used in commercial-scale biodiesel production. This is mainly due to non-optimized process design...... and a lack of available costeffective enzymes. The technology to re-use enzymes has typically proven insufficient for the processes to be competitive. However, literature data documenting the productivity of enzymatic biodiesel together with the development of new immobilization technology indicates...... that enzyme catalysts can become cost effective compared to chemical processing. This work reviews the enzymatic processing of oils and fats into biodiesel with focus on process design and economy....

  13. Enzymatic synthesis of bioactive compounds with high potential for cosmeceutical application.

    Science.gov (United States)

    Antonopoulou, Io; Varriale, Simona; Topakas, Evangelos; Rova, Ulrika; Christakopoulos, Paul; Faraco, Vincenza

    2016-08-01

    Cosmeceuticals are cosmetic products containing biologically active ingredients purporting to offer a pharmaceutical therapeutic benefit. The active ingredients can be extracted and purified from natural sources (botanicals, herbal extracts, or animals) but can also be obtained biotechnologically by fermentation and cell cultures or by enzymatic synthesis and modification of natural compounds. A cosmeceutical ingredient should possess an attractive property such as anti-oxidant, anti-inflammatory, skin whitening, anti-aging, anti-wrinkling, or photoprotective activity, among others. During the past years, there has been an increased interest on the enzymatic synthesis of bioactive esters and glycosides based on (trans)esterification, (trans)glycosylation, or oxidation reactions. Natural bioactive compounds with exceptional theurapeutic properties and low toxicity may offer a new insight into the design and development of potent and beneficial cosmetics. This review gives an overview of the enzymatic modifications which are performed currently for the synthesis of products with attractive properties for the cosmeceutical industry. PMID:27276911

  14. Detecting platform for phenolic compounds-characteristic of enzymatic electrode

    Science.gov (United States)

    Cabaj, Joanna; Chyla, Antoni; Jędrychowska, Agnieszka; Olech, Kamila; Sołoducho, Jadwiga

    2012-08-01

    We report here on simple and universal method for the highly efficient, electrolytic immobilization of tyrosinase (from Agaricus bisporus), for amperometric biosensing purposes. Tyrosinase has been successfully deposited on the surface of thin, ordered films of copolymerized derivative of thiophene (3-methylthiophene/3-thiopheneacetic acid/bis(ethylenedioxythiophene)diphenylamine). The tyrosinase retains well its activity well within the fabricated copolymer matrix. Reduction peaks, observed in cyclic voltammetry at 0.125 to +0.07 V, were attributed to the reduction of enzymatically liberated quinone species. Considering the fact, that immobilization strategy showed high efficiency, obtained results suggest that the method for phenoloxidase immobilization has a great potential for fabrication of bioelectronics' devices.

  15. An enzymatic signal amplification system for calorimetric studies of cellobiohydrolases

    DEFF Research Database (Denmark)

    Murphy, Leigh; Baumann, Martin Johannes; Borch, Kim;

    2010-01-01

    The study of cellulolytic enzymes has traditionally been carried out using endpoint measurements by quantitation of reaction products using high-performance liquid chromatography (HPLC) or overall determination of produced reducing ends. To measure catalytic activity, model substrates such as...... is heat production. This can be converted to the rate of reaction and allows direct and continuous monitoring of the hydrolysis of complex substrates. To overcome the low molar enthalpy of the hydrolysis of the glycosidic bond, which is typically on the order of −2.5 kJ mol−1, an enzymatic signal...

  16. Multifunctional modification of wool using an enzymatic process in aqueous-organic media.

    Science.gov (United States)

    Hossain, Kh M Gaffar; González, María Díaz; Lozano, Guillem Rocasalbas; Tzanov, Tzanko

    2009-04-20

    An enzymatic method using laccases for grafting the water insoluble phenolic compound lauryl gallate on wool fabric was developed. To find the compromise conditions at which the substrate is soluble while the enzyme remains active, the reaction was carried out in an 80/20 (v/v, %) aqueous-ethanol mixture, where the enzyme retains 75-80% of its activity. The enzymatic coating of wool with lauryl gallate provided in a one-step process a multifunctional textile material with antioxidant, antibacterial and water repellent properties. PMID:19428731

  17. In-vitro antioxidant and antibacterial properties of fermentatively and enzymatically prepared chicken liver protein hydrolysates.

    Science.gov (United States)

    Chakka, Ashok Kumar; Elias, Mercy; Jini, R; Sakhare, P Z; Bhaskar, N

    2015-12-01

    Protein hydrolysates were prepared from chicken liver using fermentation and enzymatic hydrolysis. The lactic acid bacteria Pediococcus acidilactici NCIM5368 was employed in the fermentation process and a commercial protease (Alcalase® 2.5) was used in enzymatic hydrolysis. Chicken liver hydrolysates prepared by fermentation (FCLH) and enzymatic hydrolysis (ECLH) revealed appreciable amounts of protein [55.85 and 61.34 %; on dry weight basis, respectively]. Fermentation and enzymatic hydrolysis resulted in 14.3 and 26.12 % of degree of hydrolysis. Total antioxidant activity, reducing power, scavenging of superoxide, 2- diphenyl-1-picrylhydrazyl (DPPH) and 2, 2-azino-bis-3-ethyl-benzthiazoline-6-sulphonic acid (ABTS) radicals were determined for both FCLH & ECLH. FCLH & ECLH showed total antioxidant activity of 0.99 and 1.13 μg AAE mg(-1) proteins, respectively; while, they scavenged 96.14 and 92.76 % of DPPH radicals respectively. FCLH showed higher ABTS radical scavenging activity (32.16 %) than ECLH (19.29 %). Superoxide anion scavenging activity of FCLH & ECLH were found to be 95.02 & 88.94 %, respectively. Residues obtained after both treatments also exhibited antioxidant activities. FCLH reported highest antagonistic activity against Listeria monocytogenes (30 mm); while, ECLH showed antibacterial activity only against Micrococcus luteus (12 mm). Both hydrolysates have the potential to be a protein rich ingredient for use in formulated foods and possible help in reduction of oxidative stress. PMID:26604378

  18. Kinetics of enzymatic hydrolysis of methyl ricinoleate

    Directory of Open Access Journals (Sweden)

    Neeharika, T. S.V.R.

    2015-12-01

    Full Text Available Ricinoleic acid is an unsaturated hydroxy fatty acid that naturally occurs in castor oil in proportions of up to 85–90%. Ricinoleic acid is a potential raw material and finds several applications in coatings, lubricant formulations and pharmaceutical areas. Enzymatic hydrolysis of castor oil is preferred over conventional hydrolysis for the preparation of ricinoleic acid to avoid estolide formation. A kinetics analysis of the enzymatic hydrolysis of Methyl Ricinoleate in the presence of Candida antarctica Lipase B was carried out in this study by varying reaction temperature (40–60 °C and enzyme concentration (2–5%. The optimal conditions were found to be 6 h reaction time, temperature 60°C, buffer to methyl ricinoleate ratio 2:1(v/w and 4% enzyme concentration to achieve a maximum conversion of 98.5%. A first order reversible reaction kinetic model was proposed to describe this reaction and a good agreement was observed between the experimental data and the model values. The effect of temperature on the forward reaction rate constant was determined by fitting data to the Arrhenius equation. The activation energy for forward reaction was found to be 14.69 KJ·mol−1.El ácido ricinoleico es un hidroxiácido insaturado que se produce naturalmente en el aceite de ricino en proporciones de hasta el 85–90%. El ácido ricinoleico es una materia prima con gran potencial y tiene aplicaciones en revestimientos, formulaciones lubricantes y en áreas farmacéuticas. Para la preparación del ácido ricinoleico se prefiere la hidrólisis enzimática del aceite de ricino a la hidrólisis convencional, para evitar la formación de estólidos. En este estudio se llevó a cabo la cinética de la hidrólisis enzimática del ricinoleato de metilo en presencia de lipasa de Candida antarctica B mediante la variación de la temperatura de reacción (40–60 °C y la concentración de la enzima (2–5%. Las condiciones óptimas de la reacción para

  19. Enzymatic hydrolysis of pretreated soybean straw

    International Nuclear Information System (INIS)

    In order to produce lactic acid, from agricultural residues such as soybean straw, which is a raw material for biodegradable plastic production, it is necessary to decompose the soybean straw into soluble sugars. Enzymatic hydrolysis is one of the methods in common use, while pretreatment is the effective way to increase the hydrolysis rate. The optimal conditions of pretreatment using ammonia and enzymatic hydrolysis of soybean straw were determined. Compared with the untreated straw, cellulose in straw pretreated by ammonia liquor (10%) soaking for 24 h at room temperature increased 70.27%, whereas hemicellulose and lignin in pretreated straw decreased to 41.45% and 30.16%, respectively. The results of infrared spectra (IR), scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis also showed that the structure and the surface of the straw were changed through pretreatment that is in favor of the following enzymatic hydrolysis. maximum enzymatic hydrolysis rate of 51.22% was achieved at a substrate concentration of 5% (w/v) at 50 deg. C and pH 4.8 using cellulase (50 fpu/g of substrate) for 36 h

  20. Starch: chemistry, microstructure, processing and enzymatic degradation

    Science.gov (United States)

    Starch is recognized as one of the most abundant and important commodities containing value added attributes for a vast number of industrial applications. Its chemistry, structure, property and susceptibility to various chemical, physical and enzymatic modifications offer a high technological value ...

  1. Enzymatic production of polysaccharides from gum tragacanth

    DEFF Research Database (Denmark)

    2014-01-01

    Plant polysaccharides, relating to the field of natural probiotic components, can comprise structures similar to human milk oligosaccharides. A method for enzymatic hydrolysis of gum tragacanth from the bush-like legumes of the genus Astragalus, using a combination of pectin hydrolases and a...

  2. Tandem and sequential multi-enzymatic syntheses

    NARCIS (Netherlands)

    B.G. Kim; J.H. Ahn; G. Sello; P. Di Gennaro; T. van Herk; A.F. Hartog; R. Wever; I. Oroz-Guinea; I. Sánchez-Moreno; E. García-Junceda; B. Wu; W. Szymanski; B.L. Feringa; D.B. Janssen; L. Villo; M. Kreen; M. Kudryashova; A. Metsala; S. Tamp; ü. Lille; T. Pehk; O. Parve; K. McClean; P. Eddowes

    2012-01-01

    This chapter contains sections titled: Production of Isorhamnetin 3-O-Glucoside in Escherichia coli Using Engineered Glycosyltransferase Multienzymatic Preparation of (−)-3-(Oxiran-2-yl)Benzoic Acid Enzymatic Synthesis of Carbohydrates from Dihydroxyacetone and Aldehydes by a One Pot Enzyme Cascade

  3. Enzymatic assay for methotrexate in erythrocytes

    DEFF Research Database (Denmark)

    Schrøder, H; Heinsvig, E M

    1985-01-01

    Methotrexate (MTX) accumulates in erythrocytes in MTX-treated patients. We present a modified enzymatic assay measuring MTX concentrations between 10 and 60 nmol/l in erythrocytes, adapted for a centrifugal analyser (Cobas Bio). About 40 patient's samples could be analysed within 1 h. The detection...

  4. Improved enzymatic production of phenolated glycerides through alkyl phenolate intermediate

    DEFF Research Database (Denmark)

    Yang, Zhiyong; Feddern, Vivian; Glasius, Marianne; Guo, Zheng; Xu, Xuebing

    2011-01-01

    This work reported a novel approach for synthesis of dihydrocaffoylated glycerides, consisting of 2 steps: enzymatic synthesis of octyl dihydrocaffeate (as a synthetic intermediate) from octanol and dihydrocaffeic acid (DHCA), and enzymatic interesterification of triglycerides with octyl dihydroc...

  5. Current-Voltage Modeling of the Enzymatic Glucose Fuel Cells

    OpenAIRE

    Vladimir Zeev Rubin

    2015-01-01

    Enzymatic fuel cells produce electrical power by oxidation of renewable energy sources. An enzymatic glucose biofuel cell uses glucose as fuel and enzymes as biocatalyst, to convert biochemical energy into electrical energy. The applications which need low electrical voltages and low currents have much of the interest in developing enzymatic fuel cells. An analytical modelling of an enzymatic fuel cell should be used, while developing fuel cell, to estimate its various parameters, to attain t...

  6. Alkali pretreated of wheat straw and its enzymatic hydrolysis

    OpenAIRE

    Lirong Han; Juntao Feng; Shuangxi Zhang; Zhiqing Ma; Yonghong Wang; Xing Zhang

    2012-01-01

    The efficiency of enzymatic hydrolysis of cellulose can be improved by various pretreatments of the substrate. In order to increase the efficiency of enzymatic saccharification of the wheat straw, we determined the effect of different pretreatments on the physical structure, chemical components and enzymatic saccharification of wheat straw. Our results showed that combination of grinding and sodium hydroxide (NaOH) treatment had high effect on the enzymatic hydrolysis of wheat straws. The opt...

  7. Soil solid materials affect the kinetics of extracellular enzymatic reactions

    Science.gov (United States)

    Lammirato, C.; Miltner, A.; Kästner, M.

    2009-04-01

    INTRODUCTION Soil solid materials affect the degradation processes of many organic compounds by decreasing the bioavailability of substrates and by interacting with degraders. The magnitude of this effect in the environment is shown by the fact that xenobiotics which are readily metabolized in aquatic environments can have long residence times in soil. Extracellular enzymatic hydrolysis of cellobiose (enzyme: beta-glucosidase from Aspergillus niger) was chosen as model degradation process since it is easier to control and more reproducible than a whole cell processes. Furthermore extracellular enzymes play an important role in the environment since they are responsible for the first steps in the degradation of organic macromolecules; beta-glucosidase is key enzyme in the degradation of cellulose and therefore it is fundamental in the carbon cycle and for soil in general. The aims of the project are: 1) quantification of solid material effect on degradation, 2) separation of the effects of minerals on enzyme (adsorption →change in activity) and substrate (adsorption →change in bioavailability). Our hypothesis is that a rate reduction in the enzymatic reaction in the presence of a solid phase results from the sum of decreased bioavailability of the substrate and decreased activity of enzyme molecules. The relative contribution of the two terms to the overall effect can vary widely depending on the chemical nature of the substrate, the properties of the enzyme and on the surface properties of the solid materials. Furthermore we hypothesize that by immobilizing the enzyme in an appropriate carrier the adsorption of enzymes to soil materials can be eliminated and that therefore immobilization can increase the overall reaction rate (activity loss caused by immobilization activity loss caused by adsorption to soil minerals). MATERIALS AND METHODS Enzymatic kinetic experiments are carried out in homogeneous liquid systems and in heterogeneous systems where solid

  8. Quantificação dos níveis endógenos de auxinas e da actividade enzimática das polifenoloxidases em oliveira (Olea europaea L. Quantification of endogenous auxin levels and polyphenoloxidase enzymatic activity in olive (Olea europaea L.

    Directory of Open Access Journals (Sweden)

    C. Serra

    2007-01-01

    Full Text Available A actividade enzimática de polifenoloxidases foi avaliada em folhas e na zona apical, média e basal de ramos de duas cultivares de oliveira (Olea europaea L., comuns no Alentejo (‘Galega vulgar’ e ‘Cobrançosa’, mostrando que a actividade enzimática nas folhas foi muito superior à encontrada em tecidos de ramos do ano. Maior actividade enzimática foi também detectada na variedade ‘Cobrançosa’ versus ‘Galega vulgar’. As condições óptimas para a determinação da actividade enzimática foram: pH= 5.5 e T= 40 ºC, com 20 mM de 4-metilcatecol em tampão acetato. Nestas condições o KM determinado foi: 2,60 e 3,48 mM com o método de Michaelis-Menten e Lineweaver-Burk, respectivamente. A melhor recuperação das auxinas AIA (ácido indol-3-acético e AIB (ácido indolbutírico em material vegetal foi conseguida através da extracção das amostras com acetona. A separação, identificação e quantificação do AIA e AIB em padrões, material vegetal dopado (tecidos de oliveira dopados com uma concentração conhecida de padrão e não dopado, foi efectuada por técnicas cromatográficas (HPLC-DAD e LCMS, mostrando os resultados taxas de recuperação superiores a 40% para o AIA e 60% para o AIB.The poliphenoloxidase enzymatic activity was evaluated in two olive cultivars (Olea europaea L. widespread in Alentejo (‘Galega vulgar and ‘Cobrançosa. Leaves and apical, medium and basal regions of the year stems were used as sample material. When compared with the different regions of the year stem, the results have shown that enzymatic activity was significantly higher in the leaves of both cultivars. Between cultivars, it was observed that ‘Cobrançosa’ presented higher enzymatic activity than ‘Galega vulgar’. The pH at 5.5 and 40 ºC temperature, using 20 mM of 4methylcatecol in acetate buffer were the optimized conditions for the enzymatic analysis. Under these conditions, the measured KM was 2,60 and 3,48 m

  9. Enzymatic biodegradation of pharmaceutical wastewater

    OpenAIRE

    Uwadiae S. E, Yerima Y, Azike R.U

    2011-01-01

    The present effort is an attempt to reduce pollution caused by the discharge of untreated wastewater (effluents) to the environment by using a low cost method. The effluent was bio-remediated using yeast and amylase as the active agents. The greater the decomposable matters present in an effluent, the greater the oxygen demand; the greater the Biological Oxygen Demand (BOD) and Chemical Oxygen Demand(COD) values, the less Dissolved Oxygen(DO) values. 10g of yeast and amylase were added to 100...

  10. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey

    2012-01-01

    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05.  Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.  

  11. Pregnancy Exercise Increase Enzymatic Antioxidant In Pregnant Women

    Directory of Open Access Journals (Sweden)

    Wagey Freddy Wagey

    2012-01-01

    Full Text Available Objectives: Pregnancy is a vulnerable condition to all kinds of "stress", resulting in changes of physiological and metabolic functions. This research aims to determine effect of exercise during pregnancy in increasing enzymatic antioxidant marked by increase of superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalase (CAT levels. Methods: Randomized pre and posttest control group design was employed in this study. A number of 66 pregnant women were recruited in this study and grouped into two groups, i.e 30 of them as control group and the rest as treatment group. Pregnancy exercise was performed to all 36 pregnant women from 20 weeks gestation on treatment group. The exercise was performed in the morning for about 30 minutes, twice a weeks. On the other hand, daily activities was sugested for control group. Student’s t-test was then applied to determine the mean different of treatment and control group with 5 % of significant value. Results: This study reveals that there were significantly higher increase of (superoxide dismutase (SOD, gluthation peroxidase (GSHPx, and catalse (CAT levels of treatment group compare to control group. These enzymatic antioxidant increase among these two group were around 1.36 mg/gHb for SOD; 1.14 IU/gHb for GSHPx; and 0.97 IU/gHb for CAT, (p < 0.05. Clinical outcomes, such as strengten of pelvic muscle and quality of life of treatment group were significantly better compared to control group (p < 0.05. Conclusions: This means that exercise during pregnancy ages of 20 weeks increase enzymatic antioxidant levels SOD, GSHPx, and CAT higher compare to control group without exercise.

  12. Atividade enzimática de isolados de rizóbia nativos da Amazônia Central crescendo em diferentes níveis de acidez Enzymatic activity of native Central Amazonian rhizobia strains grown in different levels of acidity

    Directory of Open Access Journals (Sweden)

    Arlem Nascimento de Oliveira

    2006-03-01

    Full Text Available A importância das bactérias conhecidas como rizóbia no estabelecimento de leguminosas tem sido amplamente reconhecida. Porém, poucas são as informações referentes ao perfil enzimático dessas bactérias benéficas. O estudo objetivou investigar a influência do pH do meio sólido sobre a atividade enzimática de rizóbios nativos da Amazônia Central. Essa triagem constitui o primeiro passo no processo de seleção de microorganismos benéficos, como produtores de enzimas de aplicação biotecnológica. Nesse estudo, 64 isolados de rizóbia foram testados para a produção extracelular de amilase, lipase, pectinase e protease, em meio YMA modificado. Excetuando a atividade pectinolítica, todas as outras enzimas (amilase, lipase e protease foram detectadas nos isolados investigados. Dois isolados (INPA R-975 e INPA R-926 exibiram atividades amilolíticas, lipolíticas e proteolíticas. Os índices enzimáticos amilolíticos e proteolíticos variaram significativamente entre os isolados e as condições de pH do meio de cultura. De maneira geral, as maiores atividades amilolíticas e proteolíticas foram exibidas pelos isolados INPA R-957, INPA R-915B e INPA R-991 em pH 6,5. O isolado INPA R-957 também se mostrou amilolítico e proteolítico nos pHs 5,0 e 8,0. Esse estudo mostrou que alguns rizóbios nativos da Amazônia representam fontes promissoras de amilase e protease de uso biotecnológico, sobretudo na tecnologia de alimentos.The importance of rhizobia bacteria in the establishment of legume plants has been widely recognized. However, information is scarce regarding the enzymatic profiles of these beneficial bacteria. The objective of this study was to investigate the influence of pH of solid medium on enzymatic activity of native Central Amazonian rhizobia strains. This screening constitutes the first step in selecting beneficial microorganisms as enzyme producers. In this study, 64 strains of rhizobia were screened for

  13. Enzymatic induction of supramolecular order and bioactivity

    Science.gov (United States)

    Yang, Chengbiao; Ren, Xinrui; Ding, Dan; Wang, Ling; Yang, Zhimou

    2016-05-01

    We showed in this study that enzymatic triggering is a totally different pathway for the preparation of self-assembling nanomaterials to the heating-cooling process. Because the molecules were under lower energy levels and the molecular conformation was more ordered during the enzymatic triggeration under mild conditions, nanomaterials with higher supramolecular order could be obtained through biocatalytic control. In this study, nanoparticles were obtained by an enzymatic reaction and nanofibers were observed through the heating-cooling process. We observed a distinct trough at 318 nm from the CD spectrum of a particle sample but not a fiber sample, suggesting the long range arrangement of molecules and helicity in the nanoparticles. The nanoparticles with higher supramolecular order possessed much better potency as a protein vaccine adjuvant because it accelerated the DC maturation and elicited stronger T-cells cytokine production than the nanofibers. Our study demonstrated that biocatalytic triggering is a useful method for preparing supramolecular nanomaterials with higher supramolecular order and probably better bioactivity.We showed in this study that enzymatic triggering is a totally different pathway for the preparation of self-assembling nanomaterials to the heating-cooling process. Because the molecules were under lower energy levels and the molecular conformation was more ordered during the enzymatic triggeration under mild conditions, nanomaterials with higher supramolecular order could be obtained through biocatalytic control. In this study, nanoparticles were obtained by an enzymatic reaction and nanofibers were observed through the heating-cooling process. We observed a distinct trough at 318 nm from the CD spectrum of a particle sample but not a fiber sample, suggesting the long range arrangement of molecules and helicity in the nanoparticles. The nanoparticles with higher supramolecular order possessed much better potency as a protein vaccine

  14. Effects of sonication and high-pressure carbon dioxide processing on enzymatic hydrolysis of egg white proteins

    OpenAIRE

    Knežević-Jugović Zorica D.; Stefanović Andrea B.; Žuža Milena G.; Milovanović Stoja L.; Jakovetić Sonja M.; Manojlović Verica B.; Bugarski Branko M.

    2012-01-01

    The objectives of this study were to examine the effect of sonication and high-pressure carbon dioxide processing on proteolytic hydrolysis of egg white proteins and antioxidant activity of the obtained hydrolysates. It appeared that the ultrasound pretreatment resulted in an increase in the degree of hydrolysis of the enzymatic reaction while the high-pressure carbon dioxide processing showed an inhibition effect on the enzymatic hydrolysis of egg white proteins to some extent. The ant...

  15. Temperature induced decoupling of enzymatic hydrolysis and carbon remineralization in long-term incubations of Arctic and temperate sediments

    OpenAIRE

    Robador, Alberto; Brüchert, Volker; Steen, Andrew; Arnosti, Carol

    2010-01-01

    Udgivelsesdato: 2010 Extracellular enzymatic hydrolysis of high-molecular weight organic matter is the initial step in sedimentary organic carbon degradation and is often regarded as the rate-limiting step. Temperature effects on enzyme activities may therefore exert an indirect control on carbon mineralization. We explored the temperature sensitivity of enzymatic hydrolysis and its connection to subsequent steps in anoxic organic carbon degradation in long-term incubations of sediments fr...

  16. Enzymatic hydrolysis of lignocelluloses: Identification of novel cellulase genes from filamentous fungi

    DEFF Research Database (Denmark)

    Kolasa, Marta; Ahring, Birgitte Kiær; Lübeck, Peter Stephensen;

    2010-01-01

    bonds. Cellulose can be degraded to simple sugar components by means of enzymatic hydrolysis. However, due to its complex, crystalline structure it is difficult to break it down and the cooperative action of a variety of cellulolytic enzymes is necessary. Fungi are known to have potential in production...... of a variety of cellulolytic enzymes. The aim of this work is to discover new thermostable and robust cellulolytic enzymes for improved enzymatic hydrolysis of biomass. For this purpose two screening methods are applied in different fungal strains with high cellulolytic activities: an expression...

  17. Simple enzymatic procedure for l‐carnosine synthesis: whole‐cell biocatalysis and efficient biocatalyst recycling

    OpenAIRE

    Heyland, Jan; Antweiler, Nicolai; Lutz, Jochen; Heck, Tobias; Geueke, Birgit; Kohler, Hans‐Peter E.; Blank, Lars M.; Schmid, Andreas

    2009-01-01

    Summary β‐Peptides and their derivates are usually stable to proteolysis and have an increased half‐life compared with α‐peptides. Recently, β‐aminopeptidases were described as a new enzyme class that enabled the enzymatic degradation and formation of β‐peptides. As an alternative to the existing chemical synthesis routes, the aim of the present work was to develop a whole‐cell biocatalyst for the synthesis and production of β‐peptides using this enzymatic activity. For the optimization of th...

  18. Structured Triacylglycerol of Palm-based Margarine Fat by Enzymatic Interesterification

    DEFF Research Database (Denmark)

    Ibrahim, Nuzul Amri Bin

    2008-01-01

    The effect of enzymatic interesterification of a margarine formulation containing fish oil was studied by comparing the physical characteristics of the interesterified products with the same formulation without any modification process. Based on a response surface formulation design, thirteen oil...... stronger influence than PKO was observed in the interesterified product. Interesterified products contained higher solid fat at a temperature range from 5 to 30 C than the blend. Enzymatic interesterification has an advantage compared to blending since the former led to products having a sharp melting...... immobilized enzymes, led to an improvement of the enzymatic activity in a batch reactor. An improvement was also observed in reaction by lipase AK Amano 20 mixed with Lipozyme TL IM, a combination of non-immobilized with immobilized lipases. The co-immobilization action from the carrier of the immobilized...

  19. Chemo‐enzymatic epoxidation–process options for improving biocatalytic productivity

    DEFF Research Database (Denmark)

    Hagström, Anna E. V.; Törnvall, Ulrika; Nordblad, Mathias;

    2011-01-01

    The reactor choice is crucial when designing a process where inactivation of the biocatalyst is a problem. The main bottleneck for the chemo‐enzymatic epoxidation has been found to be enzyme inactivation by the hydrogen peroxide, H2O2, substrate. In the work reported here, the effect of reaction...... at 1.5 M resulted in increased productivity, up to 76 grams of product per gram of biocatalyst with higher retention of enzyme activity. Further investigation included a multistage design that separated the enzymatic reaction and the saturation of the RME substrate with H2O2 into different vessels....... This setup showed that the reaction rate as well as enzyme inactivation is strongly dependent on the H2O2 concentration. A 20‐fold improvement in enzymatic efficiency is required for reaching an economically feasible process. This will require a combination of enzyme modification and careful process design...

  20. Enzymatic hydrolysis of corn bran arabinoxylan

    DEFF Research Database (Denmark)

    Agger, Jane

    This thesis concerns enzymatic hydrolysis of corn bran arabinoxylan. The work has focused on understanding the composition and structure of corn bran with specific interest in arabinoxylan with the main purpose of targeting enzymatic hydrolysis for increased yields. Corn bran has been used as a...... model substrate because it represents a readily available agroindustrial side product with upgrading potentials. Corn bran originates from the wet-milling process in corn starch processing, is the outmost layers of the corn kernel and is particularly rich in pentose monosaccharides comprising the major...... components of arabinoxylan. Corn bran is one of the most recalcitrant cereal byproducts with arabinoxylans of particular heterogeneous nature. It is also rich in feruloyl derived substitutions, which are responsible for extensive cross-linking between arabinoxylan molecules and thereby participate in a...

  1. Enzymatic degradation of polycaprolactone-gelatin blend

    Science.gov (United States)

    Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

    2015-04-01

    Blends of polycaprolactone (PCL), a synthetic polymer and gelatin, natural polymer offer a optimal combination of strength, water wettability and cytocompatibility for use as a resorbable biomaterial. The enzymatic degradation of PCL, gelatin and PCL-gelatin blended films was studied in the presence of lipase (Novozym 435, immobilized) and lysozyme. Novozym 435 degraded the PCL films whereas lysozyme degraded the gelatin. Though Novozym 435 and lysozyme individually could degrade PCL-gelatin blended films, the combination of these enzymes showed the highest degradation of these blended films. Moreover, the enzymatic degradation was much faster when fresh enzymes were added at regular intervals. The changes in physico-chemical properties of polymer films due to degradation were studied by scanning electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. These results have important implications for designing resorbable biomedical implants.

  2. Enzymatic Aqueous Extraction of Soybean Oil

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The procedure of enzymatic aqueous extraction of soybean oil was assessed when two-step controlled enzymatic hydrolysis was applied. With aqueous extraction of soybean oil-containing protein, the highest yield of oil was 96.1% at the optimized conditions studied. Soybean oil-containing protein was hydrolyzed and resulted in releasing part of oil. The separated protein that contained 40% oil was enriched due to its adsorption capacity of released oil, the average oil extraction yeild reached 93.5%. Then the high oil content protein was hydrolyzed again to release oil by enzyme, the oil extraction yeild was 80.4%. As a result, high quality of soybean oil was obtained and the content of total oil yield was 74.4%.

  3. Production of MAG via enzymatic glycerolysis

    Science.gov (United States)

    Jamlus, Norul Naziraa Ahmad; Derawi, Darfizzi; Salimon, Jumat

    2015-09-01

    Enzymatic glycerolysis of a medium chain methyl ester, methyl laurate was performed using lipase Candida antarctica (Novozyme 435) for 6 hours at 55°C. The percentage of components mixture of product were determined by using gas chromatography technique. The enzymatic reaction was successfully produced monolaurin (45.9 %), dilaurin (47.1 %) and trilaurin (7.0 %) respectively. Thin layer chromatography (TLC) plate also showed a good separation of component spots. Fourier transformation infra-red (FTIR) spectrum showed the presence of ester carbonyl at wavenumber 1739.99 cm-1 and hydrogen bonded O-H at 3512.03 cm-1. The product is potentially to be used as emulsifier and additive in food industry, pharmaceutical, as well as antibacterial.

  4. Process design for enzymatic peptide synthesis in near-anhydrous organic media

    NARCIS (Netherlands)

    Vossenberg, P.; Beeftink, H.H.; Stuart, M.A.C.; Tramper, J.

    2013-01-01

    This work is a case study on a process design for enzymatic peptide synthesis, which is based on and inspired by previously established data about the Alcalase-catalyzed coupling of an amino acid amide and a chemically synthesized activated N-protected amino acid carbamoylmethyl ester in near-anhydr

  5. Expression of enzymatically inactive wasp venom phospholipase A1 in Pichia pastoris

    DEFF Research Database (Denmark)

    Borodina, Irina; Jensen, Bettina M; Wagner, Tim; Hachem, Maher A; Søndergaard, Ib; Poulsen, Lars K

    2011-01-01

    in yeast Pichia pastoris, which can give high yields of correctly folded protein on defined minimal medium and secretes relatively few native proteins simplifying purification.Residual amounts of enzymatically active phospholipase A1 could be expressed, but the venom protein had a deleterious effect...

  6. Expression of enzymatically inactive wasp venom phospholipase A1 in Pichia pastoris

    DEFF Research Database (Denmark)

    Borodina, Irina; Jensen, Bettina M.; Wagner, Tim; Abou Hachem, Maher; Søndergaard, Ib; Poulsen, Lars Kærgaard

    in yeast Pichia pastoris, which can give high yields of correctly folded protein on defined minimal medium and secretes relatively few native proteins simplifying purification. Residual amounts of enzymatically active phospholipase A1 could be expressed, but the venom protein had a deleterious effect...

  7. Expression of Enzymatically Inactive Wasp Venom Phospholipase A1 in Pichia pastoris

    DEFF Research Database (Denmark)

    Borodina, Irina; Jensen, Bettina M.; Wagner, Tim; Abou Hachem, Maher; Søndergaard, Ib; Poulsen, Lars Kærgaard

    2011-01-01

    in yeast Pichia pastoris, which can give high yields of correctly folded protein on defined minimal medium and secretes relatively few native proteins simplifying purification.Residual amounts of enzymatically active phospholipase A1 could be expressed, but the venom protein had a deleterious effect...

  8. Release of Antioxidant Capacity from Five Plant Foods during a Multistep Enzymatic Digestion Protocol

    NARCIS (Netherlands)

    Papillo, V.A.; Vitaglione, P.; Graziani, G.; Gokmen, V.; Fogliano, V.

    2014-01-01

    This study aimed at elucidating the influence of food matrix on the release of antioxidant activity from five plant foods (apple, spinach, walnut, red bean, and whole wheat). To this purpose a protocol based on sequential enzymatic digestion was adopted. The total antioxidant capacity (TAC) of both

  9. Enzymatic biodegradation of pharmaceutical wastewater

    Energy Technology Data Exchange (ETDEWEB)

    Uwadiae, S.E.; Yerima, Y.; Azik, R.U. [Department of Chemical Engineering, Igbinedion University, Okada, P.M.B. 0006, Benin City, Edo State (Nigeria)

    2011-07-01

    The present effort is an attempt to reduce pollution caused by the discharge of untreated wastewater (effluents) to the environment by using a low cost method. The effluent was bio-remediated using yeast and amylase as the active agents. The greater the decomposable matters present in an effluent, the greater the oxygen demand; the greater the Biological Oxygen Demand (BOD) and Chemical Oxygen Demand(COD) values, the less Dissolved Oxygen(DO) values. 10g of yeast and amylase were added to 1000ml each of pharmaceutical effluent. 150 ml of the effluent (from the yeast and amylase) dosed was withdrawn weekly for analysis alongside with the effluent without enzymes for turbidity, DO, BOD and COD. After a period of six weeks the effluent dosed with yeast gave the highest performance followed by that dosed with amylase. The result shows that as time increases, the amount of oxygen demand reduces while the dissolved oxygen content of the effluent increases. This indicates that the yeast enzyme was able to aid remediation of the pharmaceutical effluent.

  10. Enzymatic biodegradation of pharmaceutical wastewater

    Directory of Open Access Journals (Sweden)

    Uwadiae S. E, Yerima Y, Azike R.U

    2011-07-01

    Full Text Available The present effort is an attempt to reduce pollution caused by the discharge of untreated wastewater (effluents to the environment by using a low cost method. The effluent was bio-remediated using yeast and amylase as the active agents. The greater the decomposable matters present in an effluent, the greater the oxygen demand; the greater the Biological Oxygen Demand (BOD and Chemical Oxygen Demand(COD values, the less Dissolved Oxygen(DO values. 10g of yeast and amylase were added to 1000ml each of pharmaceutical effluent. 150 ml of the effluent (from the yeast and amylase dosed was withdrawn weekly for analysis alongside with the effluent without enzymes for turbidity, DO, BOD and COD. After a period of six weeks the effluent dosed with yeast gave the highest performance followed by that dosed with amylase. The result shows that as time increases, the amount of oxygen demand reduces while the dissolved oxygen content of the effluent increases. This indicates that the yeast enzyme was able to aid remediation of the pharmaceutical effluent.

  11. Enzymatic treatment of wool: A review

    OpenAIRE

    Mojsov, Kiro; Janevski, Aco; Andronikov, Darko

    2014-01-01

    The tendency of wool to felt and shrink is mainly due to its scaly structure. The enzymatic treatment of textiles significantly improves some of their properties as well as increases their aesthetic values and comfort of use. The application of enzymes in the wool modification process was studied, and it was proven that the application of enzymes has an important influence on changes in the surface structure. However, although proteases are large molecules, their attack is not onl...

  12. ENZYMATIC DEINKING AGENTS FOR MIXED OFFICE WASTEPAPER

    Institute of Scientific and Technical Information of China (English)

    HuayuQiu; ChuanfuLiu; XiaokeMa; YingjuanFu

    2004-01-01

    This article focused on deinking agents for enzymatic deinking of MOW (mixed office wastepaper). The deinking performances of many series of surfactants were discussed at the experimental conditions, and finally some surfactants, which had good deinking effect, were selected. Then two-composed deinking agents were discussed. The deinkability of the deinking agents, e.g. deinking agents containing T-123 50% and P-10 50%, T-123 70% and O-15 30%, were better than that of the imported product.

  13. ENZYMATIC DEINKING AGENTS FOR MIXED OFFICE WASTEPAPER

    Institute of Scientific and Technical Information of China (English)

    Huayu Qiu; Chuanfu Liu; Xiaoke Ma; Yingjuan Fu

    2004-01-01

    This article focused on deinking agents for enzymatic deinking of MOW (mixed office wastepaper). The deinking performances of many series of surfactants were discussed at the experimental conditions, and finally, some surfactants, which had good deinking effect, were selected. Then two-composed deinking agents were discussed. The deinkability of the deinking agents, e.g. deinking agents containing T-123 50% and P-10 50%, T-123 70% and O-1530%, were better than that of the imported product.

  14. The Enzymatic Oxidation of Graphene Oxide

    OpenAIRE

    Kotchey, Gregg P.; Allen, Brett L.; Vedala, Harindra; Yanamala, Naveena; Kapralov, Alexander A.; Tyurina, Yulia Y.; Klein-Seetharaman, Judith; Kagan, Valerian E.; Star, Alexander

    2011-01-01

    Two-dimensional graphitic carbon is a new material with many emerging applications, and studying its chemical properties is an important goal. Here, we reported a new phenomenon – the enzymatic oxidation of a single layer of graphitic carbon by horseradish peroxidase (HRP). In the presence of low concentrations of hydrogen peroxide (~40 µM), HRP catalyzed the oxidation of graphene oxide, which resulted in the formation of holes on its basal plane. During the same period of analysis, HRP faile...

  15. Enzymatic Degradation of Ovalbumin by Various Proteases

    OpenAIRE

    Matsumoto, Kiyoshi; Yoshimaru, Tetsuro; Matsui, Toshiro; Osajima, Yutaka

    1997-01-01

    An investigation was made of the enzymatic hydrolysis of ovalbumin (OVA), a major allergen in egg white, by various acid and alkaline proteases. Protease YP-SS (acid protease) from Aspergillus niger and alcalase (alkaline protease) from BacilLus licheniformis were found to be useful for the degradation of OVA, respectively. OVA was almost totally hydrolyzed within 15 hr at 37℃ by alcalase. Alcalase acted rapidly to hydrolyze OVA, with about 90% of OVA being hydrolyzed within 30min., the react...

  16. Enzymatic Synthesis of Biobased Polyesters and Polyamides

    Directory of Open Access Journals (Sweden)

    Yi Jiang

    2016-06-01

    Full Text Available Nowadays, “green” is a hot topic almost everywhere, from retailers to universities to industries; and achieving a green status has become a universal aim. However, polymers are commonly considered not to be “green”, being associated with massive energy consumption and severe pollution problems (for example, the “Plastic Soup” as a public stereotype. To achieve green polymers, three elements should be entailed: (1 green raw materials, catalysts and solvents; (2 eco-friendly synthesis processes; and (3 sustainable polymers with a low carbon footprint, for example, (biodegradable polymers or polymers which can be recycled or disposed with a gentle environmental impact. By utilizing biobased monomers in enzymatic polymerizations, many advantageous green aspects can be fulfilled. For example, biobased monomers and enzyme catalysts are renewable materials that are derived from biomass feedstocks; enzymatic polymerizations are clean and energy saving processes; and no toxic residuals contaminate the final products. Therefore, synthesis of renewable polymers via enzymatic polymerizations of biobased monomers provides an opportunity for achieving green polymers and a future sustainable polymer industry, which will eventually play an essential role for realizing and maintaining a biobased and sustainable society.

  17. New approaches to enzymatic glycoside synthesis through directed evolution.

    Science.gov (United States)

    Kittl, Roman; Withers, Stephen G

    2010-07-01

    The expanding field of glycobiology requires tools for the synthesis of structurally defined oligosaccharides and glycoconjugates, while any potential therapeutic applications of sugar-based derivates would require access to substantial quantities of such compounds. Classical chemical approaches are not well suited for such large-scale syntheses, thus enzymatic approaches are sought. Traditional routes to the enzymatic assembly of oligosaccharides have involved the use of either Nature's own biosynthetic enzymes, the glycosyl transferases, or glycosidases run in transglycosylation mode. However, each approach has drawbacks that have limited its application. Glycosynthases are mutant glycosidases in which the catalytic nucleophile has been replaced by mutation, inactivating them as hydrolases. When used in conjunction with glycosyl fluorides of the opposite anomeric configuration to that of the substrate, these enzymes function as highly efficient transferases, frequently giving stoichiometric yields of products. Further improvements can be obtained through directed evolution of the gene encoding the enzyme in question, but this requires the ability to screen very large libraries of catalysts. In this review we survey new screening methods for the formation of glycosidic linkages using high-throughput techniques, such as FACS, chemical complementation, and robot-assisted ELISA assays. Enzymes were evolved to have higher catalytic activity with their natural substrates, to show altered substrate specificities or to be promiscuous for efficient application in oligosaccharide, glycolipid, and glycoprotein synthesis. PMID:20427037

  18. Enzymatic processes in alternative reaction media: a mini review

    Directory of Open Access Journals (Sweden)

    Mansour Ghaffari-Moghaddam

    2015-08-01

    Full Text Available Biocatalysis is a growing field in the production of fine chemicals and will most probably increase its share in the future. Enzymatic reactions are carried out under mild conditions, i.e., non-toxic solvents, low temperature and pressure, which eliminates most environmental drawbacks associated with conventional production methods. The superiority of chemo-, regio- and enantioselectivity of enzymes exhibit significant advantages over conventional catalysts for production of fine chemicals, flavors, fragrances, agrochemicals and pharmaceuticals. Enzymes can function both in aqueous and non-aqueous solvents. As a result of the growing scientific and industrial interest towards green chemistry, green solvent systems, which are mainly water, supercritical fluids, ionic liquids, fluorinated solvents, and solvent-free systems have become more popular in biocatalysis. However, the activity and selectivity of an enzyme is heavily dependent on solvent properties. In this review, various green solvents were classified and some of their influential features on enzyme activity were discussed.

  19. The enzymatic activity and genetic analysis of a family with one patient who have both systemic lupus erythematosus with secondary Sj(o)gren's syndrome and Fabry disease%系统性红斑狼疮继发干燥综合征合并Fabry病患者家系的酶活性和基因分析

    Institute of Scientific and Technical Information of China (English)

    马亚; 焦洋; 赵久良; 文煜冰; 张为民; 曾学军

    2012-01-01

    Objective To analyze the clinical information of a family with one patient who have systemic lupus erythematosus (SLE) and Fabry disease,as well as the enzymatic activity and gene mutation in these family members.Methods Clinical characteristics were collected from the proband and her family members.Peripheral blood samples from three members of this family were collected and the enzymatic activity was measured by fluorimetrie substrate assay.Genomic DNA was extracted from one male member with significantly decreased enzyme activity,the 7 exons and their flanking introns of GLA gene were amplified by PCR and directly sequenced.Results The enzyme activity of two family members was significantly decreased,the genetic analysis of the male member revealed a missense mutation in exon 2:c.334C>T (CGC>TGC)( p.R112C ).Family members except the proband had no definite evidence to support the presence of SLE.Conclusion The coexistence of SLE and Fabry disease is extremely rare.Immunological test,enzymatic activity and gene mutation analysis seem to be helpful for the differential diagnosis.%目的 分析1例系统性红斑狼疮(SLE)合并Fabry病患者家系临床资料、酶学和基因学的特点.方法 收集患者及其家系成员的临床资料,抽取患者家系中3名成员外周血,底物荧光法进行酶活性检测,并对其中1名酶活性显著降低的男性家系成员进行基因组DNA提取,对GLA基因的7个外显子和旁侧内含子的序列进行聚合酶链反应(PCR)产物直接测序,检测是否存在突变.结果 家系中2名成员外周血白细胞酶活性降低;其中的男性家系成员的基因检测显示GLA基因外显子2有错义突变:c.334C>T( CGC>TGC)(p.R112C);除先证者本人外,其他家系成员均无明确的SLE证据.结论 SLE合并Fabry病极其罕见;对于难以鉴别的患者需及时进行自身抗体及GLA酶学或者基因学的检测.

  20. Enzymatic generation of galactose-rich oligosaccharides/oligomers from potato rhamnogalacturonan I pectic polysaccharides.

    Science.gov (United States)

    Khodaei, Nastaran; Karboune, Salwa

    2016-04-15

    Potato pulp by-product rich in galactan-rich rhamnogalacturonan I (RG I) was investigated as a new source of oligosaccharides with potential prebiotic properties. The efficiency of selected monocomponent enzymes and multi-enzymatic preparations to generate oligosaccharides/oligomers from potato RG I was evaluated. These overall results of yield were dependent on the activity profile of the multi-enzymatic preparations. Highest oligo-RG I yield of 93.9% was achieved using multi-enzymatic preparation (Depol 670L) with higher hydrolytic activity toward side chains of RG I as compared to its backbone. Main oligo-RG I products were oligosaccharides with DP of 2-12 (79.8-100%), while the oligomers with DP of 13-70 comprised smaller proportion (0.0-20.2%). Galactose (58.9-91.2%, w/w) was the main monosaccharide of oligo-RG I, while arabinose represented 0.0-12.1%. An understanding of the relationship between the activity profile of multi-enzymatic preparations and the yield/DP of oligo-RG I was achieved. This is expected to provide the capability to generate galacto- and galacto(arabino) oligosaccharides and their corresponding oligomers from an abundant by-product. PMID:26616968