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Sample records for acid phosphatase isolated

  1. Human prostatic acid phosphatase directly stimulates collagen synthesis and alkaline phosphatase content of isolated bone cells

    Human prostatic acid phosphatase (hPAP) directly enhances the differentiated characteristics of isolated bone cells in vitro. This enzyme, when added to cell cultures for 24 h in vitro stimulates collagen synthesis and the production of alkaline phosphatase. The effects are dose dependent, with statistically significant effects occurring from 0.1-100 nM hPAP. Concentrations higher than 100 nM do not evoke greater effects. The maximal effect of hPAP occurs between 12 and 24 h of exposure. The cells stimulated to the greatest degree are osteoprogenitor cells and osteoblasts. Fibroblasts isolated from the same tissue show a lesser sensitivity to hPAP. hPAP has no detectable effect on cell proliferation, as measured by radiolabeled thymidine incorporation or total DNA synthesis. None of the observations reported in this work can be attributed to contaminating proteins in the hPAP preparation. hPAP was radiolabeled with 125I and was used for affinity binding and cross-linking studies. Scatchard analysis of specific binding indicated the presence of 1.0 X 10(5) high affinity binding sites/cell, with a Kd of 6.5 nM. Cross-linking studies demonstrated the presence of one 320-kDa binding complex. The pH profile and kinetic determinations of Km and maximum velocity for hPAP were similar to those previously reported, except for the finding of positive cooperativity of the substrate with the enzyme under the conditions of our assay. We believe that the direct stimulation of bone-forming cells by hPAP may contribute to the sclerotic nature of skeletal bone around sites of neoplastic prostatic metastases and that the effect of the enzyme is probably mediated by a plasma membrane receptor

  2. Effecf of pH and some cations on activity of acid phosphatase secreted from Ustilago sp. isolated from acid sulphate soil

    Chairatana Nilnond

    2007-03-01

    Full Text Available Acid phosphatase secreted from Ustilago sp. is able to hydrolyze organic phosphorus. These soil yeast microorganisms were isolated from rice roots grown in acid sulphate soil that generally contains highamount of aluminum (Al, iron (Fe and manganese (Mn ions. Therefore, the objectives of this study were to examine the effect of pH and some cations on acid phosphatase activity. Two isolates of Ustilago sp., AR101and AR102, were cultured in 100 mL of modified Pikovskaya's broth containing Na-phytate, pH 4, and acid phosphatase activity was determined at pH 2.0-7.0. Effect of Al, Fe, and Mn, including calcium (Ca ions,on growth of AR101 and AR102, secreted acid phosphatase activity, and the ability of acid phosphatase on the phosphorus release from Na-phytate by Ustilago sp. were investigated. It was found that the optimum pH for acid phosphatase activity was 3.5-4.5. The activity of acid phosphatase secreted from AR101 (3,690nmol min-1 mL-1 was remarkably higher than that from AR102 (956 nmol min-1 mL-1. Aluminum, iron, manganese and calcium ions in the medium did not affect the growth of either isolate. The activity of secretedacid phosphatase of AR101 was inhibited by Al and Ca ion, and synthesis of acid phosphatase of Ustilago sp. AR102 was possibly stimulated by Fe ion. Both AR101 and AR102 solubilized Na-phytate, resulting in therelease of P. However, some amount of released P was then precipitated with Al and Fe ions as the highly insoluble Fe- or Al- phosphate.

  3. Acid phosphatase test proves superior to standard phenotypic identification procedure for Clostridium perfringens strains isolated from water

    Ryzinska-Paier, G.; Sommer, R.; Haider, J.M.; Knetsch, S.; Frick, C.; Kirschner, A.K.T.; Farnleitner, A.H.

    2011-01-01

    Clostridium perfringens is used as an indicator for persistent faecal pollution as well as to monitor the efficacy of water treatment processes. For these purposes, differentiation between C. perfringens and other Clostridia is essential and is routinely carried out by phenotypic standard tests as proposed in the ISO/CD 6461-2:2002 (ISO_LGMN: lactose fermentation, gelatine liquidation, motility and nitrate reduction). Because the ISO_LGMN procedure is time consuming and labour intensive, the acid phosphatase test was investigated as a possible and much more rapid alternative method for confirmation. The aim of our study was to evaluate and compare confirmation results obtained by these two phenotypic methods using genotypically identified strains, what to our knowledge has not been accomplished before. For this purpose, a species specific PCR method was selected based on the results received for type strains and genotypically characterised environmental strains. For the comparative investigation type strains as well as presumptive C. perfringens isolates from water and faeces samples were used. The acid phosphatase test revealed higher percentage (92%) of correctly identified environmental strains (n = 127) than the ISO_LGMN procedure (83%) and proved to be a sensitive and reliable confirmation method. PMID:21872622

  4. Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract

    RATKO M. JANKOV

    2002-09-01

    Full Text Available An acid phosphatase from an extract of mugwort (Artemisia vulgaris pollen was purified by a factor of 48 by a combination of ion exchange and gel-chromatography. The molecular weights of the enzyme were 76 kDa and 73 kDa, determined by gel filtration on a Sephadex G-100 sf column and by SDS PAGE (under reducing and non-reducing conditions, respectively. In analytical isoelectrofocusing, the enzyme appears as two very close bands, pI at about 4.2. The optimum pH for the enzyme is 5.4. The apparent Km for p-nitrophenyl phosphate was estimated to be 0.16 mM. The purified enzyme has broad specificity, and hydrolyses p-nitrophenyl phosphate and a-naphthyl phosphate. Pyrophosphate and O-phospho-L-tyrosine were estimated to be the best substrates for this enzyme as potential in vivo substrates. The enzyme is inhibited competitively by phosphate (Ki = 1.25 mM, molybdate (Ki = 0.055 mM and pyrophosphate (Ki = 6.7 mM and non-competitively by fluoride (Ki = 9.8 mM. Metal ions such as Hg2+, Cu2+ and Zn2+ express an inhibitory effect on the enzyme, while the enzyme is slightly activated by non-ionic detergents, Tween 20 and Triton X-100. There is no change in the enzyme activity in the presence of tartrate, citrate, EDTA, 1,10-phenanthroline and sulfhydryl-group modifiers such as p-chloromercuribenzoate and N-ethylmaleimide.

  5. Phosphorus compounds, proteins, nuclease and acid phosphatase activities in isolated spinach chloroplasts

    E. Mikulska

    2015-05-01

    Full Text Available This paper deals with attempts to elaborate a simple method of spinach chloroplast isolation ensuring a high proportion of intact chloroplasts. We obtained 3 preparations of isolated chloroplasts. Several preliminary analyses of the obtained chloroplast fraction were also performed. Phosphorus compounds, total protein and the enzyme activities of RNase, DNase and GPase were determined. We found: 0,36-0,59% of RNA, 0,19-0,24% of DNA, 2,1-2,9% of phospholipids and 26-28% of protein. RNase activity was very high.

  6. Primary structure of rat secretory acid phosphatase and comparison to other acid phosphatases.

    Roiko, K; Jänne, O A; Vihko, P

    1990-05-14

    Overlapping cDNA clones encoding rat prostatic acid phosphatase (rPAP) were isolated by using two human prostatic acid phosphatase (hPAP)-encoding cDNAs to screen rat prostatic cDNA libraries. The isolated cDNAs encompassed a total of 1626 nucleotides (nt), of which 1143 nt corresponded to the protein coding sequence encoding a mature polypeptide of 350 amino acids (aa) and a 31-aa long signal peptide-like sequence. The deduced Mr of the mature rPAP was 40,599. RNA blot analysis indicated the presence of three mRNA species (4.9, 2.3 and 1.5 kb in size) in the rat prostate. The deduced aa sequences of rPAP and hPAP show 75% identity, whereas the similarity between rPAP and human lysosomal acid phosphatase (hLAP) is only 45%. Furthermore, the sequence similarity between rPAP and rat lysosomal acid phosphatase (rLAP) is 46% at the aa level. Similar to hPAP, but unlike hLAP and rLAP, the rPAP sequence lacks a membrane-anchoring domain indicating the secretory character of this phosphatase. All six cysteines present in the overlapping areas of the mature rPAP, hPAP, rLAP and hLAP proteins are positionally conserved, suggesting that these residues are important for the tertiary structure of acid phosphatases (APs). The previously reported active site residues, two arginines and one histidine, are also conserved in these APs. PMID:2373368

  7. Potentiometric assay for acid and alkaline phosphatase

    Simple potentiometric kinetic assay for evaluation of acid and alkaline phosphatase activity has been developed. Enzymatically catalyzed hydrolysis of monofluorophosphate, the simplest inorganic compound containing P-F bond, has been investigated as the basis of the assays. Fluoride ions formed in the course of the hydrolysis of this specific substrate have been detected using conventional fluoride ion-selective electrode based on membrane made of lanthanum fluoride. The key analytical parameters necessary for sensitive and selective detection of both enzymes have been assessed. Maximal sensitivity of the assays was observed at monofluorophosphate concentration near 10-3 M. Maximal sensitivity of acid phosphatase assay was found at pH 6.0, but pH of 4.8 is recommended to eliminate effects from alkaline phosphatase. Optimal pH for alkaline phosphatase assay is 9.0. The utility of the developed substrate-sensor system for determination of acid and alkaline phosphatase activity in human serum has been demonstrated

  8. Mammalian-like Purple Acid Phosphatases in Plants

    2006-01-01

    @@ Introduction Purple acid phosphatases (PAPs) comprise of a family of binuclear metal-containing hydrolases, some members of which have been isolated and characterized from animal, plant and fungal sources[1]. PAPs not only catalyze the hydrolyses of a wide range of phosphate esters and anhydrides under acidic reaction conditions,but also catalyze the generation of hydroxyl radicals in a Fenton-like reaction, by virtue of the presence of a redox-active binuclear metal center.

  9. Acid phosphatase purified from Mycoplasma fermentans has protein tyrosine phosphatase-like activity.

    Shibata, K; Noda, M.; Sawa, Y; Watanabe, T.

    1994-01-01

    Acid phosphatase purified from Mycoplasma fermentans dephosphorylated phosphotyrosine-containing lysozyme and Raytide, a peptide substrate for protein tyrosine phosphatases. The optimum pH for Raytide was about 5.5. Raytide phosphatase activity was inhibited by potassium fluoride, sodium molybdate, and sodium orthovanadate and was found to exist in some mycoplasmas.

  10. Origin and production of phosphatases in the acid Lake Gardsjoen

    Olsson, H.

    1983-01-01

    The activity of acid phosphatases was followed for one year in Lake Gardsjoen as well as in the inlet and the outlet of the lake. A budget of the phosphatases was calculated, including an estimation of the production of phosphatases. The phosphatase activity was also measured in two basins upstream of L. Gardsjoen: the north basin and the south basin of L. Stora Haestevatten. The acid phosphatase activity was very high compared with reported alkaline phosphatase activities in other lakes. About 95% of the phosphatases in L. Gardsjoen was produced in the lake, and the production was highest in early summer. Small Chrysophyceae (< 10 ..mu..m) probably produced the majority of the acid phosphatases in the investigated lakes, and accordingly could be favoured in environments with low phosphorus supply due to their ability to produce large amounts of phosphatases. 10 references, 8 figures, 2 tables.

  11. Nucleotide and amino acid sequences of human intestinal alkaline phosphatase: close homology to placental alkaline phosphatase

    A cDNA clone for human adult intestinal alkaline phosphatase (ALP) [orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1] was isolated from a λgt11 expression library. The cDNA insert of this clone is 2513 base pairs in length and contains an open reading frame that encodes a 528-amino acid polypeptide. This deduced polypeptide contains the first 40 amino acids of human intestinal ALP, as determined by direct protein sequencing. Intestinal ALP shows 86.5% amino acid identity to placental (type 1) ALP and 56.6% amino acid identity to liver/bone/kidney ALP. In the 3'-untranslated regions, intestinal and placental ALP cDNAs are 73.5% identical (excluding gaps). The evolution of this multigene enzyme family is discussed

  12. Protein tyrosine phosphatase 1B inhibitors isolated from Artemisia roxburghiana.

    Shah, Muhammad Raza; Ishtiaq; Hizbullah, Syed Muhammad; Habtemariam, Solomon; Zarrelli, Armando; Muhammad, Akhtar; Collina, Simona; Khan, Inamulllah

    2016-08-01

    Artemisia roxburghiana is used in traditional medicine for treating various diseases including diabetes. The present study was designed to evaluate the antidiabetic potential of active constituents by using protein tyrosine phosphatase 1B (PTP1B) as a validated target for management of diabetes. Various compounds were isolated as active principles from the crude methanolic extract of aerial parts of A. roxburghiana. All compounds were screened for PTP1B inhibitory activity. Molecular docking simulations were performed to investigate the mechanism behind PTP1B inhibition of the isolated compound and positive control, ursolic acid. Betulinic acid, betulin and taraxeryl acetate were the active PTP1B principles with IC50 values 3.49 ± 0.02, 4.17 ± 0.03 and 87.52 ± 0.03 µM, respectively. Molecular docking studies showed significant molecular interactions of the triterpene inhibitors with Gly220, Cys215, Gly218 and Asp48 inside the active site of PTP1B. The antidiabetic activity of A. roxburghiana could be attributed due to PTP1B inhibition by its triterpene constituents, betulin, betulinic acid and taraxeryl acetate. Computational insights of this study revealed that the C-3 and C-17 positions of the compounds needs extensive optimization for the development of new lead compounds. PMID:26118418

  13. PURIFICATION AND PARTIAL CHARACTERIZATION OF AN ACID PHOSPHATASE FROM SPIRODELA OLIGORRHIZA AND ITS AFFINITY FOR SELECTED ORGANOPHOSPHATE PESTICIDES

    An acid phosphatase from the aquatic plant Spirodela oligorrhiza (duckweed) was isolated by fast protein liquid chromatography (FPLC) and partially characterized. The enzyme was purified 1871-fold with a total yield of 40%. SDS-PAGE electrophoresis of the pure acid phosphatase ...

  14. Radiation-induced alterations in splenic acid phosphatase of pigeons

    The effect of total body ν-irradiation with sub-lethal dose (400 rad) on acid phosphatase has been studied in spleen of pigeons. The specific activity of acid phosphatase increased significantly 48 hr and 72 hr after irradiation. This increase was accompanied by a substantial reduction in per cent 'bound' activity. The histochemical observation after irradiation confirmed the result obtained by quantitative biochemical study. This increase in acid phosphatase activity may be attributed to an increased permeability of lysosomal membrane caused by damaged lymphocytes (lymphocytolysis) after ν-irradiation. (author)

  15. Revisiting histidine-dependent acid phosphatases: a distinct group of tyrosine phosphatases

    Veeramani, Suresh; Lee, Ming-Shyue; Lin, Ming-Fong

    2009-01-01

    Although classical protein tyrosine phosphatase (PTP) superfamily members are cysteine-dependent, emerging evidence shows that many acid phosphatases (AcPs) function as histidine-dependent PTPs in vivo. These AcPs dephosphorylate phospho-tyrosine substrates intracellularly and could have roles in development and disease. In contrast to cysteine-dependent PTPs, they utilize histidine, rather than cysteine, for substrate dephosphorylation. Structural analyses reveal that active site histidine, ...

  16. Plasma acid and alkaline phosphatase in patients with breast cancer.

    Nguyen, M; Bonneterre, J; Hecquet, B; Desoize, B; Demaille, A

    1991-01-01

    Acid and alkaline phosphatase were determined in 107 breast cancer patients to study their potential value in case of bone metastases. The patients were divided into 4 groups: A, patients without metastases (n = 34); B, metastatic patients without bone lesions (n = 37); C, patients with metastases in and outside of bones (n = 24), D, patients with bone-only metastases (n = 12). Tartrate resistant acid phosphatase (TR-ACP), and bone alkaline phosphatase (bone-ALP) were significantly higher in patients with metastases than in patients without. However, no difference in TR-ACP was observed between subgroups of metastatic patients. PMID:2064338

  17. Phosphatase activity of Poa pratensis seeds. I. Preliminary studies on acid phosphatase II

    I. Lorenc-Kubis

    2015-05-01

    Full Text Available Acid phosphatase (EC 3.1.3.2 was extracted with 0.1 M sodium acetate buffer pH 5.1 from Poa pratensis seeds, and separated into three fractions by chromatography on DEAE cellulose. The highest activity was found in fraction Il-b (acid phosphatase II. The activity of the enzyme was optimal at pH 4.9. It hydrolyzed p-nitrophenyl phosphate most readily among the various phosphomonoesters examined. Acid phosphatase II showed also a high activity toward β-naphtyl phosphate and phenyl phosphate, very low activity towards β-glycero phosphate, 5'-GMP and no activity with glucose-1 phosphate. The enzyme was inhibited by Ca2+ and fluoride, but activated by Mg2+. EDTA had no influence on the activity of the enzyme.

  18. Acid phosphatase and lipid peroxidation in human cataractous lens epithelium

    Vasavada Abhay

    1993-01-01

    Full Text Available The anterior lens epithelial cells undergo a variety of degenerative and proliferative changes during cataract formation. Acid phosphatase is primarily responsible for tissue regeneration and tissue repair. The lipid hydroperoxides that are obtained by lipid peroxidation of polysaturated or unsaturated fatty acids bring about deterioration of biological membranes at cellular and tissue levels. Acid phosphatase and lipid peroxidation activities were studied on the lens epithelial cells of nuclear cataract, posterior subcapsular cataract, mature cataract, and mixed cataract. Of these, mature cataractous lens epithelium showed maximum activity for acid phosphatase (516.83 moles of p-nitrophenol released/g lens epithelium and maximum levels of lipid peroxidation (86.29 O.D./min/g lens epithelium. In contrast, mixed cataractous lens epithelium showed minimum activity of acid phosphatase (222.61 moles of p-nitrophenol released/g lens epithelium and minimum levels of lipid peroxidation (54.23 O.D./min/g lens epithelium. From our study, we correlated the maximum activity of acid phosphatase in mature cataractous lens epithelium with the increased areas of superimposed cells associated with the formation of mature cataract. Likewise, the maximum levels of lipid peroxidation in mature cataractous lens epithelium was correlated with increased permeability of the plasma membrane. Conversely, the minimum levels of lipid peroxidation in mixed cataractous lens epithelium makes us presume that factors other than lipid peroxidation may also account for the formation of mixed type of cataract.

  19. Association of erythrocyte acid phosphatase phenotypes with myopia

    Himabindu P

    2005-01-01

    Full Text Available Acid phosphatase is a polymorphic nonspecific orthophosphate monoesterase which catalyses the cleaving of phosphoric acid and subsequent breakdown of several monophosphoric esters under acidic pH conditions. Acid phosphatase has a physiologic function as a flavin mononucleotide phosphatase (FMN and regulates the intracellular concentrations of flavin coenzymes that are electron carriers in the oxidative phosphorylation pathway. Myopia or nearsightedness is caused by both environmental and genetic factors. Myopic eyes when subjected to excessive oxidative stress results in retinal detachments .In the present study there is a significant elevation of AA phenotype in myopes when compared to controls. The AA phenotype is more susceptible to oxidative stress and its lower enzyme activity is known to be associated with increased intrauterine growth that further results in increased axial length in progressive myopia. The AA phenotype also confers risk for myopia development in males, early age group and cases with parental consanguinity.

  20. Lipophosphoglycan and secreted acid phosphatase of Leishmania tropica share species-specific epitopes.

    Jaffe, C L; Perez, L; Schnur, L F

    1990-06-01

    Several species-specific monoclonal antibodies (T11, T13-T15) which only react with Leishmania tropica, recognize phosphorlated carbohydrate epitopes on lipophosphoglycan and the structurally related molecule, phosphoglycan, which is shed by promastigotes into spent culture medium. During immunoaffinity isolation of [32P]orthophosphate-labeled phosphoglycan on monoclonal antibody T15 conjugated to Sepharose 4B, a high-Mr component (approx. 200,000) was co-purified. The latter material is metabolically labeled with [35S]methionine and [3H]glucosamine. This glycoprotein was separated from phosphoglycan by chromatography on lentil lectin resin. The glycoprotein exhibited a L-tatrate-sensitive acid phosphatase activity, typical of secreted acid phosphatase (EC 3.1.3.2) from Leishmania. Monospecific antibodies to Leishmania donovani-secreted acid phosphatase selectively precipitated the L. tropica enzyme from immunoaffinity purified mixtures of the two antigens, and monoclonal antibodies to lipophosphoglycan precipitate the pure enzyme. Species-specific monoclonal antibodies to L. major lipophosphoglycan also recognized both L. tropica antigens. Treatment of the acid phosphatase with periodate or phosphodiesterase I abolished binding by the monoclonal antibodies to the pure enzyme. These results demonstrate that the two major secreted glycoconjugates of Leishmania tropica, the lipophosphoglycan and the acid phosphatase, share species-specific phosphorylated carbohydrate epitope(s). PMID:1697935

  1. Prostatic acid phosphatase, purification and iodination using Iodogen

    Prostatic acid phosphatase was purified from prostatic adenomas. The procedure involved chromatography on Concanavalin A-Sepharose, DEAE-cellulose, Bio-Gel P-150 and L-tartrate-Sepharose. The purified phosphatase hydrolyzed p-nitrophenyl phosphate at a rate of 270 μmol.mg-1.min-1 (250C) and showed homogeneity upon polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The final prostatic acid phosphatase preparation was pure and the antisera were monospecific as judged by the highly-sensitive technique of crossed immunoelectrophoresis. Of the procedures evaluated for the radioiodination of the purified enzyme with iodine 125, oxidation with Iodogen was found to give the best radioiodinated product, to be used in radioimmunoassay. (Auth.)

  2. Biocatalysis with Sol-Gel Encapsulated Acid Phosphatase

    Kulkarni, Suhasini; Tran, Vu; Ho, Maggie K.-M.; Phan, Chieu; Chin, Elizabeth; Wemmer, Zeke; Sommerhalter, Monika

    2010-01-01

    This experiment was performed in an upper-level undergraduate biochemistry laboratory course. Students learned how to immobilize an enzyme in a sol-gel matrix and how to perform and evaluate enzyme-activity measurements. The enzyme acid phosphatase (APase) from wheat germ was encapsulated in sol-gel beads that were prepared from the precursor…

  3. The effect of potassium iodide on the production of acid phosphatase by Sporothrix schenckii

    P. S. Grover

    2003-06-01

    Full Text Available The present study was undertaken to find out the in vitro effect of potassium iodide (KI on the production of acid phosphatase by fully characterized strain of S.schenckii isolated from a patient of Cutaneous Sporotrichosis. The enzyme acid phosphatase was estimated during the 3 phases of growth of S.schenckii, without and with three concentrations of KI incorporated in the culture medium. In the control and in the test proper, with various concentrations of KI, no adverse effect of KI was observed on the production of acid phosphatase in early and mid log phase of fungal growth. Whereas in the exponential phase in test proper, there was a statistical significant decrease in the enzyme production with 0.8% and 3.2% of KI. The low activity at 0.8% and 3.2% KI indicates that KI has inhibitory effect on the growth of S.schenckii and has led to decrease in the activity of the enzyme. (Med J Indones 2003; 12: 65-8 Keywords: S.schenckii, acid phosphatase, potassium iodide

  4. The development of determining human prostatic acid phosphatase by radioimmunoassay

    We purified human prostatic acid phosphatase (hPAP) from prostatic tissues by affinity chromatography, DEAE cellulose and gel filtration and also examined physicochemical properties of highly purified PAP. We developed a double-antibody radioimmunoassay for hPAP in serum, with use of antiserum raised in rabbit against highly purified PAP. The antiserum did not cross react with acid phosphatase from platelets and red blood cells. Experimental detail are outlined to assess the reproducibility and reliability of the method under various conditions. The upper limit of the serum PAP levels in the present assay was set at 3.0 ng/ml by 162 determinations of samples. The serum PAP levels of 2 untreated patients with prostatic carcinoma were higher than 3.0 ng/ml and 39 patients with benign prostatic hyperplasia were an average value of 1.9 ng/ml. (author)

  5. Monomeric Tartrate Resistant Acid Phosphatase Induces Insulin Sensitive Obesity

    Lång, Pernilla; van Harmelen, Vanessa; Rydén, Mikael; Kaaman, Maria; Parini, Paolo; Carneheim, Claes; Cassady, A. Ian; Hume, David A.; Andersson, Göran; Arner, Peter

    2008-01-01

    Background Obesity is associated with macrophage infiltration of adipose tissue, which may link adipose inflammation to insulin resistance. However, the impact of inflammatory cells in the pathophysiology of obesity remains unclear. Tartrate resistant acid phosphatase (TRAP) is an enzyme expressed by subsets of macrophages and osteoclasts that exists either as an enzymatically inactive monomer or as an active, proteolytically processed dimer. Principal Findings Using mice over expressing TRAP...

  6. Molecular cloning and sequence analysis of cDNA encoding human prostatic acid phosphatase.

    Vihko, P; Virkkunen, P; Henttu, P; Roiko, K; Solin, T; Huhtala, M L

    1988-08-29

    lambda gt11 clones encoding human prostatic acid phosphatase (PAP) (EC 3.1.3.2) were isolated from human prostatic cDNA libraries by immunoscreening with polyclonal antisera. Sequence data obtained from several overlapping clones indicated that the composite cDNAs contained the complete coding region for PAP, which encodes a 354-residue protein with a calculated molecular mass of 41,126 Da. In the 5'-end, the cDNA codes for a signal peptide of 32 amino acids. Direct protein sequencing of the amino-terminus of the mature protein and its proteolytic fragments confirmed the identity of the predicted protein sequence. PAP has no apparent sequence homology to other known proteins. However, both the cDNA clones coding for human placental alkaline phosphatase and PAP have an alu-type repetitive sequence about 900 nucleotides downstream from the coding region in the 3'-untranslated region. Two of our cDNA clones differed from others at the 5'-ends. RNA blot analysis indicated mRNA of 3.3 kb. We are continuing to study whether acid phosphatases form a gene family as do alkaline phosphatases. PMID:2842184

  7. Acid phosphatase complex from the freshwater snail Viviparus viviparus L. under standard conditions and intoxication by cadmium ions.

    Tsvetkov, I L; Popov, A P; Konichev, A S

    2003-12-01

    Acid phosphatases differing in both subcellular localization and substrate specificity were isolated for the first time from the liver of the freshwater snail Viviparus viviparus L. by preparative isoelectrofocusing. One of five characterized phosphatases is highly specific to ADP and the others can hydrolyze (at variable rate) a series of natural substrates. A scheme is proposed for the involvement of the studied phosphatases in carbohydrate metabolism. We have also studied some peculiarities of the effect of Cd2+ in vitro and in vivo on the activities of individual components of the acid phosphatase complex and corresponding changes in metabolism of the freshwater snail as a new test-object allowing the estimation of toxicity in water. PMID:14756629

  8. Testicular acid phosphatase induces odontoblast differentiation and mineralization.

    Choi, Hwajung; Kim, Tak-Heun; Yun, Chi-Young; Kim, Jung-Wook; Cho, Eui-Sic

    2016-04-01

    Odontoblasts differentiate from dental mesenchyme during dentin formation and mineralization. However, the molecular mechanisms controlling odontoblast differentiation remain poorly understood. Here, we show that expression of testicular acid phosphatase (ACPT) is restricted in the early stage of odontoblast differentiation in proliferating dental mesenchymal cells and secretory odontoblasts. ACPT is expressed earlier than tissue-nonspecific alkaline phosphatase (TNAP) and partly overlaps with TNAP in differentiating odontoblasts. In MDPC-23 odontoblastic cells, expression of ACPT appears simultaneously with a decrease in β-catenin activity and is abolished with the expression of Phex and Dsp. Knockdown of ACPT in MDPC-23 cells stimulates cell proliferation together with an increase in active β-catenin and cyclin D1. In contrast, the overexpression of ACPT suppresses cell proliferation with a decrease in active β-catenin and cyclin D1. Expression of TNAP, Osx, Phex and Dsp is reduced by knockdown of ACPT but is enhanced by ACPT overexpression. When ACPT is blocked with IgG, alkaline phosphatase activity is inhibited but cell proliferation is unchanged regardless of ACPT expression. These findings suggest that ACPT inhibits cell proliferation through β-catenin-mediated signaling in dental mesenchyme but elicits odontoblast differentiation and mineralization by supplying phosphate during dentin formation. Thus, ACPT might be a novel candidate for inducing odontoblast differentiation and mineralization for dentin regeneration. PMID:26547858

  9. Direct Electrochemistry of Porcine Purple Acid Phosphatase (Uteroferrin)

    Bernhardt, Paul V; Schenk, Gerhard; Wilson, Gregory J.

    2004-01-01

    Cyclic voltammetry of the non-heme diiron enzyme porcine purple acid phosphatase (uteroferrin, Uf) has been reported for the first time. Totally reversible one-electron oxidation responses (FeIII-FeII f FeIII-FeIII) are seen both in the absence and in the presence of weak competitive inhibitors phosphate and arsenate, and dissociation constants of these oxoanion complexes formed with uteroferrin in its oxidized state (Ufo) have been determined. The effect of pH on the redox potent...

  10. Acid phosphatase localization in neurons of Bulla gouldiana (Gastropoda: Opisthobranchia.

    Robles, L J; Fisher, S K

    1975-01-01

    The organization of the ganglia and the ultrastructure of the neurons of Bulla gouldiana are similar to those described for other molluscs. Acid phosphatase positive reactions were found in the large pigmented granules, small dense bodies, multivesicular bodies, and Golgi lamellae and associated vesicles. The small dense bodies and multivesicular bodies may be stages in the formation of the larger pigmented granules which are interpreted as lysosomes. Comparison is made between the pigmented granules in Bulla and the lipofuscin bodies of vertebrate neurons. The possible involvement of these pigmented granules in the hyperpolarization of Bulla and Aplysia neurons to light is discussed. PMID:1122539

  11. Study on prostatic acid phosphatase (PAP) immunoradiometric assay kit

    This coat-antibody-count PAP IRMA is a solid-phase immunoradiometric assay based on two strains of monoclonal antibodies, designed for the quantitative measurement of prostatic acid phosphatase (PAP) in serum. The minimal detectable concentration is 0.1 μg/L. The intra and inter coefficients of variation are 8.8%-9.6% and 7.7%-12.3%, respectively. The recovery is 96.3%-105.0% and the range of detection is 2.5-200.0 μg/L

  12. Cathepsin D-mediated yolk protein degradation is blocked by acid phosphatase inhibitors.

    Fialho, Eliane; Nakamura, Angelica; Juliano, Luiz; Masuda, Hatisaburo; Silva-Neto, Mário A C

    2005-04-15

    Vitellin (VT) is a lipoglycophosphoprotein stored inside the eggs of every oviparous organism during oogenesis. In the blood-sucking bug Rhodnius prolixus, VT is deposited inside growing oocytes together with two acid hydrolases: acid phosphatase (AP) and cathepsin D (CD). Egg fertilization triggers AP activity and VT proteolysis in vivo [Insect Biochem. Mol. Biol. 2002 (32) 847]. Here, we show that CD is the main protease targeting VT proteolysis during egg development. CD activity in total egg homogenates is blocked by the classical aspartyl protease inhibitor, pepstatin A. Surprisingly, AP inhibitors such as NaF, Na+/K+ tartrate, and inorganic phosphate also block VT proteolysis, whereas this effect is not observed when tyrosine phosphatase inhibitors such as vanadate and phenylarsine oxide or an inhibitor of alkaline phosphatases such as levamisole are used in a VT proteolysis assay. NaF concentrations that block isolated AP activity do not affect the activity of partially purified CD. Therefore, a specific repressor of VT proteolysis must be dephosphorylated by AP in vivo. In conclusion, these results demonstrate for the first time that acid hydrolases act cooperatively to promote yolk degradation during egg development in arthropods. PMID:15797237

  13. Isolation and characterization of a neutral phosphatase from wheat seedlings

    A neutral phosphatase was purified to homogeneity from wheat seedlings. The enzyme was a monomeric glycoprotein exhibiting a molecular weight of 35,000, frictional ratio of 1.22, Stokes' radius of 26 A, and sedimentation coefficient of 3.2 S. That the enzyme was a glycoprotein was surmised from its chromatographic property on Concanavalin A-Sepharose column. The phosphatase activity was assayed using either fructose-2,6-bisphosphate or p-nitrophenyl phosphate as substrate. The phosphatase activity was not affected by high concentrations of chelating agents and did not require the addition of Mg+2 or Ca+2 for its activity. Molybdate, orthovanadate, Zn+2, and Hg+2 were all potent inhibitors of the phosphatase activity. The inhibition by Hg+2 was reversed by dithiothreitol. The enzyme activity was stimulated by Mn+2 about 2-fold. On the other hand, 3-phosphoglycerate, fructose-6-P and Pi as well as polyamines inhibited the enzyme activity. The ability of the neutral phosphatase to dephosphorylate protein phosphotyrosine was also investigated. The phosphotyrosyl-substrates, such as [32P] phosphotyrosyl-poly(Glu, Tyr)n, -alkylated bovine serum albumin, -angiotensin-1, and -band 3 of erythrocytes, were all substrates of the phosphatase. On the other hand, the enzyme had no activity toward protein phosphoserine and protein phosphothreonine

  14. Prostatic acid phosphatase is the main acid phosphatase with 5'-ectonucleotidase activity in the male mouse saliva and regulates salivation.

    Araujo, César L; Quintero, Ileana B; Kipar, Anja; Herrala, Annakaisa M; Pulkka, Anitta E; Saarinen, Lilli; Hautaniemi, Sampsa; Vihko, Pirkko

    2014-06-01

    We have previously shown that in addition to the well-known secreted isoform of prostatic acid phosphatase (sPAP), a transmembrane isoform exists (TMPAP) that interacts with snapin (a SNARE-associated protein) and regulates the endo-/exocytic pathways. We have also shown that PAP has 5'-ectonucleotidase and thiamine monophosphatase activity and elicits antinociceptive effects in mouse models of chronic inflammatory and neuropathic pain. Therefore, to determine the physiological role of PAP in a typical exocrine organ, we studied the submandibular salivary gland (SMG) of PAP(-/-) and wild-type C57BL/6J mice by microarray analyses, microRNA sequencing, activity tests, immunohistochemistry, and biochemical and physiological analyses of saliva. We show that PAP is the main acid phosphatase in the wild-type male mouse saliva, accounting for 50% of the total acid phosphatase activity, and that it is expressed only in the granular convoluted tubules of the SMGs, where it is the only 5'-ectonucleotidase. The lack of PAP in male PAP(-/-) mice was associated with a significant increase in the salivation volume under secretagogue stimulation, overexpression of genes related to cell proliferation (Mki67, Aurkb, Birc5) and immune response (Irf7, Cxcl9, Ccl3, Fpr2), and upregulation of miR-146a in SMGs. An increased and sustained acinar cell proliferation was detected without signs of glandular hyperplasia. Our results indicate that in PAP(-/-) mice, SMG homeostasis is maintained by an innate immune response. Additionally, we suggest that in male mice, PAP via its 5'-ectonucleotidase activity and production of adenosine can elicit analgesic effects when animals lick their wounds. PMID:24717577

  15. Cervical acid phosphatase detection: A guide to abnormal cells in cytology smear screening for cervical cancer

    Deb Prabal; Iyer Venkateswaran; Bhatla Neerja; Markovic O; Verma Kusum

    2008-01-01

    Background: Cervical acid phosphatase-Papanicolaou (CAP-PAP) test has recently been described for detection of acid phosphatase enzyme in abnormal squamous cells, and has been proposed as a biomarker-based technology for the screening of cervical cancer. Materials and Methods: Eighty-one consecutive cervical smears were subjected to routine Papanicolaou (Pap) staining as well as CAP-PAP, which combined cytochemical staining for acid phosphatase with modified Pap stain. Statistical evaluation ...

  16. High degree of homology between primary structure of human lysosomal acid phosphatase and human prostatic acid phosphatase.

    Peters, C; Geier, C; Pohlmann, R; Waheed, A; von Figura, K; Roiko, K; Virkkunen, P; Henttu, P; Vihko, P

    1989-02-01

    Alignment of the amino-acid sequences of the human lysosomal acid phosphatase (LAP) and human prostatic acid phosphatase (PAP) yielded an extensive homology between the two mature polypeptide chains. In the overlapping part, which extends over the entire PAP sequence and the N-terminal 90% of the LAP sequence, the identity is 49.1%. The LAP has an additional C-terminal sequence, which is encoded by the last exon of the LAP gene. This sequence contains the transmembrane domain of LAP, which is lacking in the secretory PAP. All six cysteine residues as well as 20 out of 27 (LAP) and 26 (PAP) proline residues present in the overlapping part of the proteins are conserved, suggesting that they are involved in stabilization of the tertiary structure of both proteins. Only two out of 8 N-glycosylation sites in LAP and 3 in PAP are conserved, suggesting that the dense N-glycosylation of LAP is related to its function in lysosomes. PMID:2706086

  17. Lowering of phytic acid content by enhancement of phytase and acid phosphatase activities during sunflower germination

    Juliana da Silva Agostini; Rosicler Balduíno Nogueira; Elza Iouko Ida

    2010-01-01

    The objective of this work was to investigate the germination of hybrid sunflowers BRS191 and C11 as a means of lowering phytic acid (PA) content by enhancing the activity of endogenous phytase and acid phosphatase. The concentration of PA in hybrid sunflower achenes varied from 2.16 to 2.83g/100g of sample (p < 0.05). The phytase and acid phosphatase activities of sunflowers BRS191 and C11 were the highest on the 4th and 5th days of germination, respectively, with the release of the phosphor...

  18. Biochemical characterization and subcellular localization of the red kidney bean purple acid phosphatase

    Phosphatases are known to play a crucial role in phosphate turnover in plants. However, the exact role of acid phosphatases in plants has been elusive because of insufficient knowledge of their in vivo substrate and subcellular localization. We investigated the biochemical properties of a purple acid phosphatase isolated from red kidney bean (Phaseolus vulgaris) (KBPAP) with respect to its substrate and inhibitor profiles. The kinetic parameters were estimated for five substrates. We used 31P nuclear magnetic resonance to investigate the in vivo substrate of KBPAP. Chemical and enzymological estimation of polyphosphates and ATP, respectively, indicated the absence of polyphosphates and the presence of ATP in trace amounts in the seed extracts. Immunolocalization using antibodies raised against KBPAP was unsuccessful because of the nonspecificity of the antiserum toward glycoproteins. Using histoenzymological methods with ATP as a substrate, we could localize KBPAP exclusively in the cell walls of the peripheral two to three rows of cells in the cotyledons. KBPAP activity was not detected in the embryo. In vitro experiments indicated that pectin, a major component of the cell wall, significantly altered the kinetic properties of KBPAP. The substrate profile and localization suggest that KBPAP may have a role in mobilizing organic phosphates in the soil during germination

  19. Method for isolating nucleic acids

    Hurt, Jr., Richard Ashley; Elias, Dwayne A.

    2015-09-29

    The current disclosure provides methods and kits for isolating nucleic acid from an environmental sample. The current methods and compositions further provide methods for isolating nucleic acids by reducing adsorption of nucleic acids by charged ions and particles within an environmental sample. The methods of the current disclosure provide methods for isolating nucleic acids by releasing adsorbed nucleic acids from charged particles during the nucleic acid isolation process. The current disclosure facilitates the isolation of nucleic acids of sufficient quality and quantity to enable one of ordinary skill in the art to utilize or analyze the isolated nucleic acids for a wide variety of applications including, sequencing or species population analysis.

  20. Phosphoglycosylation of a secreted acid phosphatase from Leishmania donovani.

    Lippert, D N; Dwyer, D W; Li, F; Olafson, R W

    1999-06-01

    The secreted acid phosphatase (SAcP) of L.donovani is a heterogeneous glycoprotein that displays a wide array of N- and O-linked glycosylations. The O-linked sugars are of particular interest due to their similarity to the phosphoglycan structures of the major lipophosphoglycan surface antigen and released phosphoglycan (Turco et al., 1987; Greis et al., 1992). This study describes a structural analysis of the SAcP O-linked glycosylations using mass spectroscopy, amino acid sequencing, and enzymatic carbohydrate sequencing. Analysis of glycan chain lengths and peptide glycosylation site distribution was performed, revealing that the average O-linked structure was approximately 32 repeat units in length. Amino acid sequence analysis of glycosylated peptides showed that phosphoglycosylations did not occur randomly but were localized to specific serine residues within an array of degenerate serine/threonine-rich repeat sequences localized in the C-terminus. No evidence was obtained for modification of threonine residues. The observed pattern suggested that a consensus sequence may exist for localization of phosphoglycan structures. PMID:10336996

  1. Diagnostic value of prostatic acid phosphatase as determined by radioimmunoassay

    Serum concentrations of prostatic acid phosphatase (PAP) were determined with 4 different radioimmunoassays and with the standard enzymatic method (p-nitrophenylphosphate) in 35 patients with prostatic carcinoma. Staging of localized tumors was based on histopathological evaluation after radial prostatectomy and pelvic lymphnode dissection (pTsub(1-3), pN0). In tumor lesions Tsub(1-2) N0 M0 elevated PAP-serum concentrations were found by RIA-determination in only one patient. Increased PAP serum levels were observed in 43-78% of carcinomas stage T3 N0 M0 and in 54-83% in stage Tsub(2-4) Nsub(x) M1 tumors, depending on the test kit used for the PAP determination. Concentrations for PAP obtained with the 4 different RIA-kits used, varied significantly and thus are not comparable. No false positive results were observed in sera of 9 patients with benign prostatic hyperplasia. Elevated PAP serum levels were found in a significantly higher frequency when determined by radioimmunoassay than by the enzymatic method. The results clearly indicate, that PAP is of no value for early recognition of carcinoma of the prostate even when measured by radioimmunoassay. However, the RIA-method seems to be of clinical importance in estimating the course of advanced local and metastasizing carcinoma of the prostate. (orig.)

  2. Radioimmunoassay for human prostatic acid phosphatase: Pt.4

    After PAP RIA has been established, serum prostatic acid phosphatase concentration was measured in 40 healthy males, 20 healthy females, 57 patients with benign prostatic hyperplasia, 20 patients with prostate cancers at various stages, 11 patients with cancers after prostatectomy or orchiectomy, and 36 patients with cancers other than prostate cancer. An upper cutoff value was calculated from the x + 2S of healthy males, which yielded a value of 2.2 μg/L of serum. More male patients with cancers other than prostate cancer had serum PAP values of less than 2.2 g/L. 91% (52/57) of BPH patients had normal value, 9% (5/57) exhibited elevated serum PAP levels. If cutoff the limit of x + 2S, the calculated value from 57 BPH patients was used, i.e. 3.0 μg/L, as hormal limit, the false positives presented by BPH were almost eliminated. 95% (1/20) patients with prostate cancers demonstrated an evidently elevated PAP, only one of those patients had a normal PAP value. After prostatectomy of prostate cancer, PAP declined to normal range or near upper cutoff value. The values obtained by this PAP assay were able to distinguish patients with prostate cancer from those with benign prostatic hyperplasia, and the course of disease could be monitored by the assay. Intraindividual sequential studies of PAP could be used to evaluate therapeutic response and prognosis

  3. Synthesis of functionalized fluorescent gold nanoclusters for acid phosphatase sensing

    Sun, Jian; Yang, Fan; Yang, Xiurong

    2015-10-01

    A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by introducing an alkaline aqueous solution of MUA into the GSH-Au+ complexes or AuNC@GSH solution. Subsequently, a reliable AuNC@GSH/MUA-based real-time assay of acid phosphatase (ACP) is established for the first time, inspired by the selective coordination of Fe3+ with surface ligands of AuNCs, the higher binding affinity between the pyrophosphate ion (PPi) and Fe3+, and the hydrolysis of PPi into orthophosphate by ACP. Our fluorescent chemosensor can also be applied to assay ACP in a real biological sample and, furthermore, to screen the inhibitor of ACP. This report paves a new avenue for synthesizing AuNCs based on either the bottom-up reduction or top-down etching method, establishing real-time fluorescence assays for ACP by means of PPi as the substrate, and further exploring the sensing applications of fluorescent AuNCs.A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by

  4. Isolation, Purification and Characterization of Acid Phosphatase from Cilantro%芫荽酸性磷酸酶的提取、纯化及酶学性质研究

    王丹; 万骥; 傅婷; 唐云明

    2015-01-01

    采用硫酸铵分级沉淀、CM-Sepharose离子交换层析、Superdex-200凝胶过滤层析法,从新鲜芫荽中分离纯化出电泳纯的酸性磷酸酶(acid phosphatase,ACP).该酶的酶活回收率为14.20%、纯化倍数为238.60、酶比活力为295.87 U/mg、亚基分子质量约为53.8 kD;芫荽ACP酶学性质研究结果表明:该酶的最适反应温度为55℃,在50℃以下时较稳定,因此该酶对温度较敏感;该酶的最适反应pH值为5.8,在pH 4.0~7.0之间较稳定,表明该酶耐受于酸性环境;芫荽ACP的对硝基苯酚磷酸二钠km值为0.63 mmol/L,表明该酶与底物对硝基苯酚磷酸二钠具有较高的亲和力;甲醇、乙醇、异丙醇、抗坏血酸、草酸、Cu2+、Pb2+、Ag+对该酶具有强烈的抑制作用;Mg2+、Mn2+、Ba2+、K+对该酶具有一定的激活作用.

  5. Purification and Characterization of Two Extracellular Alkaline Phosphatases from a Psychrophilic Arthrobacter Isolate

    Prada, P; Brenchley, J E

    1997-01-01

    Two extracellular, heat-labile alkaline phosphatases were purified from a psychrophilic Arthrobacter isolate, D10. The enzymes were active over different pH ranges, used distinct substrates, and had different kinetic properties. Each enzyme reacted specifically to its own antibody during immunoblot analysis. One had both monophosphatase and diesterase activities.

  6. Tartrate resistant acid phosphatase in the immune and nervous system : Distribution and pathophysiological implications

    Lång, Pernilla

    2007-01-01

    Tartrate resistant acid phosphatase (TRAP) belongs to the family of purple acid phosphatases (PAP). It is a glycoprotein synthesized as a monomer with low enzyme activity containing a redox active diiron centre in the active site. Post-translational proteolytic processing of this monomer into a dimeric protein increases the enzyme activity. Traditionally, TRAP has been used as a marker for bone resorbing cells but the biological function of TRAP is still not fully elucidated...

  7. Effects of multivalent cations on cell wall-associated acid phosphatase activity

    Tu, S.I.; Brouillette, J.N.; Nagahashi, G.; Kumosinski, T.F.

    1988-09-01

    Primary cell walls, free from cytoplasmic contamination were prepared from corn (Zea mays L.) roots and potato (Solanum tuberosum) tubers. After EDTA treatment, the bound acid phosphatase activities were measured in the presence of various multivalent cations. Under the conditions of minimized Donnan effect and at pH 4.2, the bound enzyme activity of potato tuber cell walls (PCW) was stimulated by Cu/sup 2 +/, Mg/sup 2 +/, Za/sup 2 +/, and Mn/sup 2 +/; unaffected by Ba/sup 2 +/, Cd/sup 2 +/, and Pb/sup 2 +/; and inhibited by Al/sup 3 +/. The bound acid phosphatase of PCW was stimulated by a low concentration but inhibited by a higher concentration of Hg/sup 2 +/. On the other hand, in the case of corn root cells walls (CCW), only inhibition of the bound acid phosphatase by Al/sup 3 +/ and Hg/sup 2 +/ was observed. Kinetic analyses revealed that PCW acid phosphatase exhibited a negative cooperativity under all employed experimental conditions except in the presence of Mg/sup 2 +/. In contrast, CCW acid phosphatase showed no cooperative behavior. The presence of Ca/sup 2 +/ significantly reduced the effects of Hg/sup 2 +/ or Al/sup 3 +/, but not Mg/sup 2 +/, to the bound cell wall acid phosphatases. The salt solubilized (free) acid phosphatases from both PCW and CCW were not affected by the presence of tested cations except for Hg/sup 2 +/ or Al/sup 3 +/ which caused a Ca/sup 2 +/-insensitive inhibition of the enzymes. The induced stimulation or inhibition of bound acid phosphatases was quantitatively related to cation binding in the cell wall structure.

  8. Acrylamide gel electrophoresis of proteins, acid phosphatases and RN-ases from three potato varieties

    A. Kubicz; E. Wieczorek; B. Morawiecka

    2015-01-01

    Studies on variety differences in the protein and acid phosphatase patterns as well as ribunuclease activity distribution were carried out by disc electrophoresis on saline extracts of three varieties of the potato Solanum tuberosum (L.). The protein bands varied in number, position and relative abundance. One main zone of the acid phosphatase activity was detected consisting of 2-3 electrophoretically different bands. Variety differences were concerned with the number and relative abundance ...

  9. Effects of precipitation on soil acid phosphatase activity in three successional forests in southern China

    W. Huang; Liu, J; Zhou, G.; Zhang, D; Deng, Q

    2011-01-01

    Phosphorus (P) is often a limiting nutrient for plant growth in tropical and subtropical forests. Global climate change has led to alterations in precipitation in the recent years, which inevitably influences P cycling. Soil acid phosphatase plays a vital role in controlling P mineralization, and its activity reflects the capacity of organic P mineralization potential in soils. In order to study the effects of precipitation on soil acid phosphatase activity, an experiment with precipitation t...

  10. Prostatic Acid Phosphatase Is Expressed in Peptidergic and Nonpeptidergic Nociceptive Neurons of Mice and Rats

    Taylor-Blake, Bonnie; Zylka, Mark J.

    2010-01-01

    Thiamine monophosphatase (TMPase, also known as Fluoride-resistant acid phosphatase or FRAP) is a classic histochemical marker of small- to medium-diameter dorsal root ganglia (DRG) neurons and has primarily been studied in the rat. Previously, we found that TMPase was molecularly identical to Prostatic acid phosphatase (PAP) using mice. In addition, PAP was expressed in a majority of nonpeptidergic, isolectin B4-binding (IB4+) nociceptive neurons and a subset of peptidergic, calcitonin gene-...

  11. Binuclear Metal Centers in Plant Purple Acid Phosphatases: Fe-Mn in Sweet Potato and Fe-Zn in Soybean

    Schenk, Gerhard; Ge, Yubin; Carrington, Lyle E; Wynne, Ceridwen J.; Searle, Iain R.; Carroll, Bernard J; Hamilton, Susan E.; de Jersey, John

    1999-01-01

    Purple acid phosphatases comprise a family of binuclear metal-containing acid hydrolases, representatives of which have been found in animals, plants, and fungi. The goal of this study was to characterize purple acid phosphatases from sweet potato tubers and soybean seeds and to establish their relationship with the only well-characterized plant purple acid phosphatase, the FeIII–ZnII-containing red kidney bean enzyme. Metal analysis indicated the presence in the pu...

  12. Inhibition of acid, alkaline, and tyrosine (PTP1B) phosphatases by novel vanadium complexes.

    McLauchlan, Craig C; Hooker, Jaqueline D; Jones, Marjorie A; Dymon, Zaneta; Backhus, Emily A; Greiner, Bradley A; Dorner, Nicole A; Youkhana, Mary A; Manus, Lisa M

    2010-03-01

    In the course of our investigations of vanadium-containing complexes for use as insulin-enhancing agents, we have generated a series of novel vanadium coordination complexes with bidentate ligands. Specifically we have focused on two ligands: anthranilate (anc(-)), a natural metabolite of tryptophan, and imidizole-4-carboxylate (imc(-)), meant to mimic naturally occurring N-donor ligands. For each ligand, we have generated a series of complexes containing the V(III), V(IV), and V(V) oxidation states. Each complex was investigated using phosphatase inhibition studies of three different phosphatases (acid, alkaline, and tyrosine (PTP1B) phosphatase) as prima facia evidence for potential use as an insulin-enhancing agent. Using p-nitrophenyl phosphate as an artificial phosphatase substrate, the levels of inhibition were determined by measuring the absorbance of the product at 405nm using UV/vis spectroscopy. Under our experimental conditions, for instance, V(imc)(3) appears to be as potent an inhibitor of alkaline phosphatase as sodium orthovanadate when comparing the K(cat)/K(m) term. VO(anc)(2) is as potent an inhibitor of acid phosphatase and tyrosine phosphatase as the Na(3)VO(4). Thus, use of these complexes can increase our mechanistic understanding of the effects of vanadium in vivo. PMID:20071031

  13. Members of a unique histidine acid phosphatase family are conserved amongst a group of primitive eukaryotic human pathogens.

    Shakarian, Alison M; Joshi, Manju B; Yamage, Mat; Ellis, Stephanie L; Debrabant, Alain; Dwyer, Dennis M

    2003-03-01

    Recently, we identified and characterized the genes encoding several distinct members of the histidine-acid phosphatase enzyme family from Leishmania donovani, a primitive protozoan pathogen of humans. These included genes encoding the heavily phosphorylated/glycosylated, tartrate-sensitive, secretory acid phosphatases (Ld SAcP-1 and Ld SAcP-2) and the unique, tartrate-resistant, externally-oriented, surface membrane-bound acid phosphatase (Ld MAcP) of this parasite. It had been previously suggested that these enzymes may play essential roles in the growth, development and survival of this organism. In this report, to further examine this hypothesis, we assessed whether members of the L. donovani histidine-acid phosphatase enzyme family were conserved amongst other pathogenic Leishmania and related trypanosomatid parasites. Such phylogenetic conservation would clearly indicate an evolutionary selection for this family of enzymes and strongly suggest and support an important functional role for acid phosphatases to the survival of these parasites. Results of pulsed field gel electrophoresis and Southern blotting showed that homologs of both the Ld SAcPs and Ld MAcP were present in each of the visceral and cutaneous Leishmania species examined (i.e. isolates of L. donovani, L. infantum, L. tropica, L. major and L. mexicana, respectively). Further, results of enzyme assays showed that all of these organisms expressed both tartrate-sensitive and tartrate-resistant acid phosphatase activities. In addition, homologs of both the Ld SAcPs and Ld MAcP genes and their corresponding enzyme activities were also identified in two Crithidia species (C. fasciculata and C. luciliae) and in Leptomonas seymouri. In contrast, Trypanosoma brucei, Trypanosoma cruzi and Phytomonas serpens had only very-low levels of such enzyme activities. Cumulatively, results of this study showed that homologs of the Ld SAcPs and Ld MAcP are conserved amongst all pathogenic Leishmania sps. suggesting

  14. Golgi-mediated post-translational processing of secretory acid phosphatase by Leishmania donovani promastigotes.

    Bates, P A; Hermes, I; Dwyer, D M

    1990-03-01

    Monensin, an inhibitor of Golgi function, was used to investigate the role of this cell compartment in the glycosylation of Leishmania donovani promastigote secretory acid phosphatase (EC 3.1.3.2). Monensin-treated cells demonstrated morphological changes in the Golgi complex and secreted enzyme with an altered electrophoretic mobility: two discrete bands of approximately 95 and 110 kDa were found, as compared to the heterodisperse nature of the enzyme from untreated controls. Chemical deglycosylation by mild acid hydrolysis resulted in a similar effect on the electrophoretic mobility of purified extracellular enzyme. Acid phosphatase was also treated with N-glycosidase F (EC 3.5.1.52) to remove N-linked oligosaccharides. The altered lectin-binding properties of the enzyme after these two treatments demonstrated that an unusual type of galactose-containing acid-labile carbohydrate was present in secretory acid phosphatase in addition to the N-linked oligosaccharides. Further, experiments with 32P-labelled enzyme indicated that phosphodiester bonds were the structural component responsible for the sensitivity of this carbohydrate to mild acid hydrolysis. Cumulatively, these results demonstrated that a novel form of Golgi-mediated posttranslational modification had occurred to the secretory acid phosphatase presumably by the addition of an acid-labile phosphoglycan. PMID:2320058

  15. Vanadate inhibition of fungal phyA and bacterial appA2 histidine acid phosphatases

    The fungal PhyA protein, which was first identified as an acid optimum phosphomonoesterase (EC 3.1.3.8), could also serve as a vanadate haloperoxidase (EC 1.11.1.10) provided the acid phosphatase activity is shutdown by vanadate. To understand how vanadate inhibits both phytate and pNPP degrading ac...

  16. Crystallization of a newly discovered histidine acid phosphatase from Francisella tularensis

    A histidine acid phosphatase from the CDC Category A pathogen F. tularensis has been crystallized in space group P41212, with unit-cell parameters a = 61.96, c = 210.78 Å. A 1.75 Å resolution data set was collected at Advanced Light Source beamline 4.2.2. Francisella tularensis is a highly infectious bacterial pathogen that is considered by the Centers for Disease Control and Prevention to be a potential bioterrorism weapon. Here, the crystallization of a 37.2 kDa phosphatase encoded by the genome of F. tularensis subsp. holarctica live vaccine strain is reported. This enzyme shares 41% amino-acid sequence identity with Legionella pneumophila major acid phosphatase and contains the RHGXRXP motif that is characteristic of the histidine acid phosphatase family. Large diffraction-quality crystals were grown in the presence of Tacsimate, HEPES and PEG 3350. The crystals belong to space group P41212, with unit-cell parameters a = 61.96, c = 210.78 Å. The asymmetric unit is predicted to contain one protein molecule, with a solvent content of 53%. A 1.75 Å resolution native data set was recorded at beamline 4.2.2 of the Lawrence Berkeley National Laboratory Advanced Light Source. Molecular-replacement trials using the human prostatic acid phosphatase structure as the search model (28% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of F. tularensis histidine acid phosphatase will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative

  17. Effects of precipitation on soil acid phosphatase activity in three successional forests in southern China

    W. Huang

    2011-07-01

    Full Text Available Phosphorus (P is often a limiting nutrient for plant growth in tropical and subtropical forests. Global climate change has led to alterations in precipitation in the recent years, which inevitably influences P cycling. Soil acid phosphatase plays a vital role in controlling P mineralization, and its activity reflects the capacity of organic P mineralization potential in soils. In order to study the effects of precipitation on soil acid phosphatase activity, an experiment with precipitation treatments (no precipitation, natural precipitation and doubled precipitation in three successional forests in southern China was carried out. The three forests include Masson pine forest (MPF, coniferous and broad-leaved mixed forest (MF and monsoon evergreen broad-leaved forest (MEBF. Results showed that driven by seasonality of precipitation, changes in soil acid phosphatase activities coincided with the seasonal climate pattern, with significantly higher values in the wet season than in the dry season. Soil acid phosphatase activities were closely linked to forest successional stages, with enhanced values in the later stages of forest succession. In the dry season, soil acid phosphatase activities in the three forests showed a rising trend with increasing precipitation treatments. In the wet season, soil acid phosphatase activity was depressed by no precipitation treatment in the three forests. However, doubled precipitation treatment exerted a significantly negative effect on it only in MEBF. These results indicate that the potential transformation rate of organic P might be more dependent on water in the dry season than in the wet season. A decrease in organic P turnover would occur in the three forests if there was a drought in a whole year in the future. More rainfall in the wet season would also be adverse to organic P turnover in MEBF due to its high soil moisture.

  18. Acid phosphatase-1 1, a molecular marker tightly linked to root-knot nematode resistance in tomato.

    Aarts, J.M.M.J.G.

    1993-01-01

    Root knot nematode resistance in tomato is a genetic trait which is determined by a single dominant gene ( Mi ) on chromosome 6 of tomato. Information about the mRNA or protein product is completely lacking, which precludes the cloning of Mi by conventional strategies based on gene expression. However, an acid phosphatase-1 allozyme marker ( Aps-1 1 ) is known, which shows tight genetic linkage to the root knot nematode resistance trait. With a view to isolating Mi nucleotide sequences by a p...

  19. Effects of sub-lethal dose of gamma-irradiation on levels of acid phosphatase in cerebellum of pigeons

    The changes in the activities of acid phosphatase in the sham-irradiated and γ-irradiated cerebellum of pigeons have been studied both biochemically as well as histochemically after 400 rads. The specific activity of acid phosphatase decreased significantly after 48h and 72h of irradiation. The histochemical observations following total body irradiation confirmed the results obtained by quantitative biochemical studies. (author)

  20. Free Fatty Acids Inhibit Protein Tyrosine Phosphatase 1B and Activate Akt

    Eisuke Shibata

    2013-09-01

    Full Text Available Background/Aims: Accumulating evidence has suggested that free fatty acids (FFAs interact with protein kinases and protein phosphatases. The present study examined the effect of FFAs on protein phosphatases and Akt. Methods: Activities of protein phosphatase 1 (PP1, protein phosphatase 2A (PP2A, and protein tyrosine phosphatase 1B (PTP1B were assayed under the cell-free conditions. Phosphorylation of Akt was monitored in MSTO-211H human malignant pleural mesothelioma cells without and with knocking-down phosphatidylinositol 3 kinase (PI3K or 3-phosphoinositide-dependent protein kinase-1 (PDK1. Results: In the cell-free assay, unsaturated FFAs (uFFAs such as oleic, linoleic and linolenic acid and saturated FFAs (sFFAs such as stearic, palmitic, myristic, and behenic acid markedly reduced PTP1B activity, with the potential for uFFAs greater than that for sFFAs. All the investigated sFFAs inhibited PP2A activity, but otherwise no inhibition was obtained with uFFAs. Both uFFAs and sFFAs had no effect on PP1 activity. Oleic acid phosphorylated Akt both on Thr308 and Ser473, while stearic acid phosphorylated Akt on Thr308 alone. The effects of oleic and stearic acid on Akt phosphorylation were abrogated by the PI3K inhibitor wortmannin or the PDK1 inhibitor BX912 and also by knocking-down PI3K or PDK1. Conclusion: The results of the present study indicate that uFFAs and sFFAs could activate Akt through a pathway along a PI3K/PDK1/Akt axis in association with PTP1B inhibition.

  1. Crystallization of a newly discovered histidine acid phosphatase from Francisella tularensis

    Felts, Richard L.; Reilly, Thomas J.; Calcutt, Michael J.; Tanner, John J.

    2005-01-01

    A histidine acid phosphatase from the CDC Category A pathogen F. tularensis has been crystallized in space group P41212, with unit-cell parameters a = 61.96, c = 210.78 Å. A 1.75 Å resolution data set was collected at Advanced Light Source beamline 4.2.2.

  2. Lipid phosphate phosphatases regulate lysophosphatidic acid production and signaling in platelets: studies using chemical inhibitors of lipid phosphate phosphatase activity.

    Smyth, Susan S; Sciorra, Vicki A; Sigal, Yury J; Pamuklar, Zehra; Wang, Zuncai; Xu, Yong; Prestwich, Glenn D; Morris, Andrew J

    2003-10-31

    Blood platelets play an essential role in ischemic heart disease and stroke contributing to acute thrombotic events by release of potent inflammatory agents within the vasculature. Lysophosphatidic acid (LPA) is a bioactive lipid mediator produced by platelets and found in the blood and atherosclerotic plaques. LPA receptors on platelets, leukocytes, endothelial cells, and smooth muscle cells regulate growth, differentiation, survival, motility, and contractile activity. Definition of the opposing pathways of synthesis and degradation that control extracellular LPA levels is critical to understanding how LPA bioactivity is regulated. We show that intact platelets and platelet membranes actively dephosphorylate LPA and identify the major enzyme responsible as lipid phosphate phosphatase 1 (LPP1). Localization of LPP1 to the platelet surface is increased by exposure to LPA. A novel receptor-inactive sn-3-substituted difluoromethylenephosphonate analog of phosphatidic acid that is a potent competitive inhibitor of LPP1 activity potentiates platelet aggregation and shape change responses to LPA and amplifies LPA production by agonist-stimulated platelets. Our results identify LPP1 as a pivotal regulator of LPA signaling in the cardiovascular system. These findings are consistent with genetic and cell biological evidence implicating LPPs as negative regulators of lysophospholipid signaling and suggest that the mechanisms involve both attenuation of lysophospholipid actions at cell surface receptors and opposition of lysophospholipid production. PMID:12909631

  3. Transmembrane prostatic acid phosphatase (TMPAP interacts with snapin and deficient mice develop prostate adenocarcinoma.

    Ileana B Quintero

    Full Text Available The molecular mechanisms underlying prostate carcinogenesis are poorly understood. Prostatic acid phosphatase (PAP, a prostatic epithelial secretion marker, has been linked to prostate cancer since the 1930's. However, the contribution of PAP to the disease remains controversial. We have previously cloned and described two isoforms of this protein, a secretory (sPAP and a transmembrane type-I (TMPAP. The goal in this work was to understand the physiological function of TMPAP in the prostate. We conducted histological, ultra-structural and genome-wide analyses of the prostate of our PAP-deficient mouse model (PAP(-/- with C57BL/6J background. The PAP(-/- mouse prostate showed the development of slow-growing non-metastatic prostate adenocarcinoma. In order to find out the mechanism behind, we identified PAP-interacting proteins byyeast two-hybrid assays and a clear result was obtained for the interaction of PAP with snapin, a SNARE-associated protein which binds Snap25 facilitating the vesicular membrane fusion process. We confirmed this interaction by co-localization studies in TMPAP-transfected LNCaP cells (TMPAP/LNCaP cells and in vivo FRET analyses in transient transfected LNCaP cells. The differential gene expression analyses revealed the dysregulation of the same genes known to be related to synaptic vesicular traffic. Both TMPAP and snapin were detected in isolated exosomes. Our results suggest that TMPAP is involved in endo-/exocytosis and disturbed vesicular traffic is a hallmark of prostate adenocarcinoma.

  4. The genomic complement of purple acid phosphatase phytases in the Triticeae

    Madsen, Claus Krogh; Dionisio, Giuseppe; Holme, Inger;

    2011-01-01

    , PAPhy_b promoters contain elements typical of gibberellic acid induced germination related hydrolases. PAPhy_a promoters in contrast possess elements known from storage protein promoters. **Dionisio G, Madsen CK, Holm PB, Welinder KG, Jørgensen M, Stoger E, Arcalis E, Brinch-Pedersen H. Cloning and...... Characterization of Purple Acid Phosphatase Phytases from Wheat (Triticum aestivum L.), Barley (Hordeum vulgare L.), Maize (Zea maize L.) and Rice (Oryza sativa L.). Plant Physiol. 2011a, Jan 10.[Epub ahead of print]...

  5. Lowering of phytic acid content by enhancement of phytase and acid phosphatase activities during sunflower germination

    Juliana da Silva Agostini

    2010-08-01

    Full Text Available The objective of this work was to investigate the germination of hybrid sunflowers BRS191 and C11 as a means of lowering phytic acid (PA content by enhancing the activity of endogenous phytase and acid phosphatase. The concentration of PA in hybrid sunflower achenes varied from 2.16 to 2.83g/100g of sample (p O objetivo deste trabalho foi investigar a germinação de girassóis híbridos BRS 191 e C11 com finalidade de reduzir o teor de AF e aumentar as atividades de phytases e fosfatases endógenas. A concentração do AF nos aquênios de girassóis híbridos variou de 2,16 a 2,83 g /100g de amostra (p< 0,005. As atividades de fitases e fosfatases de girassóis BRS191 e C11 foram elevadas no 4º e 5º dia de germinação, respectivamente, com liberação do fósforo necessário para o desenvolvimento da semente. Estes resultados indicam que o AF do girassol hibrido reduz e a atividade de phytase aumenta em períodos distintos da germinação, possibilitando assim a aplicação desta enzima no controle do teor de AF em cereais, melhorando o seu valor nutricional.

  6. Study on alkaline and acid phosphatase activity in acute uranium intoxication

    The protective potential of diethyl barbituric acid sodium salt is studied, in comparison with that of acetazolamide, on kidneys under acute uranium intoxication. Experiments involved rats given intraperitoneal injections with uranyl acetate on 12 successive days up to a total dose of 0.5, 2.0 or 7.0 mg/kg. The resulting effects are measured by chemical assays of serum and urine for alkaline and acid phosphatase and histochemical assays for phosphatase activities in kidneys, kinetics being followed over a 30-day period after total dose administration. Protection of kidneys from toxic uranium effects was found to be of about the same degree with sodium diethyl barbiturate as with acetazolamide. (A.B.)

  7. Acrylamide gel electrophoresis of proteins, acid phosphatases and RN-ases from three potato varieties

    A. Kubicz

    2015-05-01

    Full Text Available Studies on variety differences in the protein and acid phosphatase patterns as well as ribunuclease activity distribution were carried out by disc electrophoresis on saline extracts of three varieties of the potato Solanum tuberosum (L.. The protein bands varied in number, position and relative abundance. One main zone of the acid phosphatase activity was detected consisting of 2-3 electrophoretically different bands. Variety differences were concerned with the number and relative abundance of these bands. RNase activity was detected in 4 main zones, in some of them additional subbands were visible. Differences between the three examined varieties were reflected in the occurence of the particular activity zones or their subbands.

  8. Increased Tartrate-Resistant Acid Phosphatase Expression in Osteoblasts and Osteocytes in Experimental Osteoporosis in Rats

    Solberg, Lene B.; Brorson, Sverre-Henning; Stordalen, Gunhild A.; Bækkevold, Espen S; Andersson, Göran; Reinholt, Finn P.

    2014-01-01

    Tartrate-resistant acid phosphatase (TRAP) is known as an osteoclast marker, but osteoblasts and osteocytes in the vicinity of bone remodeling sites also express TRAP. Cell culture studies suggest that osteoblasts endocytose osteoclastic TRAP for inactivation. To evaluate whether changes in osteoclast activity could alter TRAP expression in osteoblasts and/or osteocytes in vivo, we studied the ovariectomized and vitamin D-deficient rat (Ovx-D) and rats healing from rickets. Bone sections were...

  9. High acid phosphatase level in the gingival tissues of periodontitis subjects

    Pushparani, D. S.

    2015-01-01

    Aim: Periodontitis is one of the major problems slowly progressing and could affect 70% of the global population. The prevalence of periodontitis differs from mild to moderate forms of race and geographic region. The aim of this study is to determine the acid phosphatase (ACP) activity in the gingival tissues of periodontitis subjects. In this study, the activity of ACP in the gingival tissue of subjects with periodontitis was examined. Materials and Methods: A total of 30 subjects were selec...

  10. Nitrate reductase and acid phosphatase activities as affected by inorganic phosphate in corn roots

    Marie Kummerova; Józef Buczek

    2014-01-01

    The deficieny of inorganic phosphate in nutrient solution reduces by about 50 per cent NO3- absorption in corn seedlings, it decreases both in vitro and in vivo nitrate reductase (NR) activity, as well the potential and actual NR level and has a very weak effect on NR induction. Acid phosphatases activities increase in corn roots when the plants are grown in nutrient solution without phosphorus. We suggest that inorganic phosphate is required mainly for maintenance of NR activity rather, than...

  11. Optimal level of purple acid phosphatase5 is required for maintaining complete resistance to Pseudomonas syringae

    Ravichandran, Sridhar; Stone, Sophia L.; Benkel, Bernhard; Zhang, Junzeng; Berrue, Fabrice; Prithiviraj, Balakrishnan

    2015-01-01

    Plants possess an exceedingly complex innate immune system to defend against most pathogens. However, a relative proportion of the pathogens overcome host's innate immunity and impair plant growth and productivity. We previously showed that mutation in purple acid phosphatase (PAP5) lead to enhanced susceptibility of Arabidopsis to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Here, we report that an optimal level of PAP5 is crucial for mounting complete basal re...

  12. Expression, purification and crystallization of an atypical class C acid phosphatase from Mycoplasma bovis

    Methods for the expression, purification and crystallization of the class C acid phosphatase from M. bovis are reported. This enzyme is atypical in that it is nearly 20 kDa larger than other known class C acid phosphatases. Class C acid phosphatases (CCAPs) are 25–30 kDa bacterial surface proteins that are thought to function as broad-specificity 5′,3′-nucleotidases. Analysis of the newly published complete genome sequence of Mycoplasma bovis PG45 revealed a putative CCAP with a molecular weight of 49.9 kDa. The expression, purification and crystallization of this new family member are described here. Standard purification procedures involving immobilized metal-ion affinity chromatography and ion-exchange chromatography yielded highly pure and crystallizable protein. Crystals were grown in sitting drops at room temperature in the presence of PEG 3350 and HEPES buffer pH 7.5 and diffracted to 2.3 Å resolution. Analysis of diffraction data suggested a primitive monoclinic space group, with unit-cell parameters a = 78, b = 101, c = 180 Å, β = 92°. The asymmetric unit is predicted to contain six molecules, which are likely to be arranged as three dimers

  13. Effect of gamma-irradiation on nucleic acids, proteins, respiration and phosphatase activity of carrot callus cultures

    Callus tissue cultures were subjected to 60Co qamma irradiation at 0.5 Krad and analysed for nucleic acids, proteins, respiration rate and phosphatase activity on 0, 10, 20 and 30 days. The RNA contents and respiratory rates were enhanced as a result of irradiation. The RNA contents were reduced than their non-irradiated counterparts. The acid phosphatase activity was enhanced immediately after irradiation, declined on 10th and 20th day and more thereafter. (author)

  14. Alkaline and Acid Phosphatase Activity in Blood Plasma of Chickens Irradiated by Low dose Gamma Radiation

    Petar, K.; Marinko, V.; Saveta, M.; Miljenko, S.

    2004-07-01

    In our previous paper (Kraljevic et, al, 2000; Kraljevic et al 2002) we showed that the growth of the chickens hatched from eggs irradiated with 0.15 Gy gamma-rays before incubation was significantly higher than in controls during the fattening period (1-42 days). The concentration of total protein, glucose and cholesterol in the blood plasma of the same chickens was also significantly changed. In this paper an attempt was made to determine the effect of irradiation of eggs by low dose ionizing radiation before incubation upon activity of alkaline and acid phosphatase in the blood plasma of chickens hatched from irradiated eggs. The eggs of heavy breeding chickens were irradiated by dose of 0.15 Gy gamma radiation (60 Co) before incubation. Along with the chickens which were hatched from irradiated eggs, there was a control group of chickens hatched from nonirradiated eggs. All other conditions were the same for both groups. After hatching, blood samples were taken from the wing vein on days 1, 3, 5, 6, 10, 20, 30 and 42. The activity of both enzymes was determined spectrophotometrically by using Boehring Mannheim GmbH optimized kits. the activity of alkaline phosphatase in blood plasma was decreased on days 42, and the activity of acid phosphatase in the blood plasma of the same chickens was increased on day 42. Obtained results confirm our early obtained results that low dose of gamma radiation has effects upon metabolic processes in the chickens hatched from eggs irradiated before incubation. (Author)

  15. Alkaline and Acid Phosphatase Activity in Blood Plasma of Chickens Irradiated by Low dose Gamma Radiation

    In our previous paper (Kraljevic et, al, 2000; Kraljevic et al 2002) we showed that the growth of the chickens hatched from eggs irradiated with 0.15 Gy gamma-rays before incubation was significantly higher than in controls during the fattening period (1-42 days). The concentration of total protein, glucose and cholesterol in the blood plasma of the same chickens was also significantly changed. In this paper an attempt was made to determine the effect of irradiation of eggs by low dose ionizing radiation before incubation upon activity of alkaline and acid phosphatase in the blood plasma of chickens hatched from irradiated eggs. The eggs of heavy breeding chickens were irradiated by dose of 0.15 Gy gamma radiation (60 Co) before incubation. Along with the chickens which were hatched from irradiated eggs, there was a control group of chickens hatched from nonirradiated eggs. All other conditions were the same for both groups. After hatching, blood samples were taken from the wing vein on days 1, 3, 5, 6, 10, 20, 30 and 42. The activity of both enzymes was determined spectrophotometrically by using Boehring Mannheim GmbH optimized kits. the activity of alkaline phosphatase in blood plasma was decreased on days 42, and the activity of acid phosphatase in the blood plasma of the same chickens was increased on day 42. Obtained results confirm our early obtained results that low dose of gamma radiation has effects upon metabolic processes in the chickens hatched from eggs irradiated before incubation. (Author)

  16. Cloning and Characterization of a Novel Purple Acid Phosphatase Gene (MtPAP1) from Medicago truncatula Barrel Medic

    2006-01-01

    A novel purple acid phosphatase gene (MtPAP1) was isolated from the model legume Medicago truncatula Barrel Medic. The cDNA was 1 698 bp in length with an open reading frame (ORF) of 1 398 bp capable of encoding an N-terminal signal peptide of 23 amino acids. The transcripts of MtPAP1 were mainly detected in leaves under high-phosphate conditions, whereas under low-phosphate conditions the transcript level was reduced in leaves and increased in roots, with the strongest hybridization signal detected in roots. A chimeric gene construct fusing MtPAP1 and GFPwas made in which the fusion was driven by the CaMV35S promoter. Transgenlc Arabidopsis plants carrying the chimeric gene constructs showed that the fusion protein was mainly located at the apoplast based on confocal microscopic analysis, showing that MtPAP1 could be secreted to the outside of the cell directed by the signal peptide at the N-terminal. The coding region of MtPAP1 without signal peptide was inserted into the prokaryotic expression vector pET-30a (+) and overexpressed in Escherlchia coll BL21(DE3). The acid phosphatase (APase) proteins extracted from bacterial culture were found largely based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. An enzyme activity assay demonstrated that the APase activity in the transformed bacteria was 3.16-fold higher than that of control. The results imply that MtPAP1 functions to improve phosphorus acquisition in plants under conditions of phosphorus (P) stress.

  17. Acute inhibition of hepatic glucose-6-phosphatase does not affect gluconeogenesis but directs gluconeogenic flux toward glycogen in fasted rats - A pharmacological study with the chlorogenic acid derivative S4048

    van Dijk, TH; van der Sluijs, FH; Wiegman, CH; Baller, JFW; Gustafson, LA; Burger, HJ; Herling, AW; Kuipers, F; Meijer, AJ; Reijngoud, DJ

    2001-01-01

    Effects of acute inhibition of glucose-6-phosphatase activity by the chlorogenic acid derivative S4048 on hepatic carbohydrate fluxes were examined in isolated rat hepatocytes and in vivo in rats. Fluxes were calculated using tracer dilution techniques and mass isotopomer distribution analysis in pl

  18. Crystal structure and immunogenicity of the class C acid phosphatase from Pasteurella multocida

    Singh, Harkewal; Malinski, Thomas J.; Reilly, Thomas J.; Henzl, Michael T.; Tanner, John J.

    2011-01-01

    Pasteurella multocida is a pathogen of veterinary and medical importance. Here, we report the 1.85 Å resolution crystal structure of the class C acid phosphatase from this organism (denoted rPmCCAP). The structure shows that rPmCCAP exhibits the same haloacid dehalogenase fold and dimeric assembly as the class C enzyme from Haemophilus influenzae. Formation of the dimer in solution is demonstrated using analytical ultracentrifugation. The active site is devoid of a magnesium ion due to the pr...

  19. Tyrosine Phosphatase TpbA of Pseudomonas aeruginosa Controls Extracellular DNA via Cyclic Diguanylic Acid Concentrations

    Ueda, Akihiro; Wood, Thomas K.

    2010-01-01

    Inactivating the tyrosine phosphatase TpbA of Pseudomonas aeruginosa PA14 induces biofilm formation by 150-fold via increased production of the second messenger cyclic diguanylic acid (c-di-GMP). Here, we show the tpbA mutation reduces extracellular DNA (eDNA) and that increased expression of tpbA increases eDNA; hence, eDNA is inversely proportional to c-di-GMP concentrations. Mutations in diguanylate cyclases PA0169, PA4959, and PA5487 and phosphodiesterase PA4781 corroborate this trend. Th...

  20. Amino acid sequence of the cold-active alkaline phosphatase from Atlantic cod (Gadus morhua)

    Asgeirsson, Bjarni; Nielsen, Berit Noesgaard; Højrup, Peter

    2003-01-01

    Atlantic cod is a marine fish that lives at low temperatures of 0-10 degrees C and contains a cold-adapted alkaline phosphatase (AP). Preparations of AP from either the lower part of the intestines or the pyloric caeca area were subjected to proteolytic digestion, mass spectrometry and amino acid...... sequencing by Edman degradation. The primary structure exhibits greatest similarity to human tissue non-specific AP (80%), and approximately 30% similarity to AP from Escherichia coli. The key residues required for catalysis are conserved in the cod AP, except for the third metal binding site, where cod AP...

  1. Insulin controls subcellular localization and multisite phosphorylation of the phosphatidic acid phosphatase, lipin 1.

    Harris, Thurl E; Huffman, Todd A; Chi, An; Shabanowitz, Jeffrey; Hunt, Donald F; Kumar, Anil; Lawrence, John C

    2007-01-01

    Brain, liver, kidney, heart, and skeletal muscle from fatty liver dystrophy (fld/fld) mice, which do not express lipin 1 (lipin), contained much less Mg(2+)-dependent phosphatidic acid phosphatase (PAP) activity than tissues from wild type mice. Lipin harboring the fld(2j) (Gly(84) --> Arg) mutation exhibited relatively little PAP activity. These results indicate that lipin is a major PAP in vivo and that the loss of PAP activity contributes to the fld phenotype. PAP activity was readily detected in immune complexes of lipin from 3T3-L1 adipocytes, where the protein was found both as a microsomal form and a soluble, more highly phosphorylated, form. Fifteen phosphorylation sites were identified by mass spectrometric analyses. Insulin increased the phosphorylation of multiple sites and promoted a gel shift that was due in part to phosphorylation of Ser(106). In contrast, epinephrine and oleic acid promoted dephosphorylation of lipin. The PAP-specific activity of lipin was not affected by the hormones or by dephosphorylation of lipin with protein phosphatase 1. However, the ratio of soluble to microsomal lipin was markedly increased in response to insulin and decreased in response to epinephrine and oleic acid. The results suggest that insulin and epinephrine control lipin primarily by changing localization rather than intrinsic PAP activity. PMID:17105729

  2. Partial Purification and Properties of an Acid Phosphatase from Pearl Oyster Pinctada Fucata

    柴云峰; 谢莉萍; 张荣庆

    2003-01-01

    Acid phosphatases (ACPs) are marker enzymes for the detection of lysosomes in cell fractions.However, ACPs in sea creatures are less studied than those on land.An acid phosphatase was partially purified from pearl oyster Pinctada fucata by chromatography on Sephadex G-150 and Con A-Sepharose 4B.The specific activity was 1719 U*mg-1 and with optimum pH (5.0) and temperature (60℃).The enzyme was strongly inhibited competitively by product analog WO3-4 and MoO3-4, but less inhibited by product analog AsO3-4.The enzyme could also be strongly inhibited by heavy metal ions, such as Ag+ and Cu2+, but was not affected by Pb2+.High concentrations of ethanol (64%) and NaF (10-3 mol·L-1) could inhibit the enzyme while low concentration of NaF (<10-4 mol·L-1) could slightly activate the enzyme.Other haloids (Cl-, Br-, I-) and EDTA did not have any effect on this enzyme, while tartrate and some chemical modification reagents (bromoacetic acid, formaldehyde and dithiothreitol) could inhibit the enzyme.It is concluded that the properties of the enzyme are different from many fresh water mollusks.

  3. Association of acid phosphatase locus 1*C allele with the risk of cardiovascular events in rheumatoid arthritis patients

    Teruel, María; Martín, J. E.; González-Juanatey, C.; López-Mejias, Raquel; Miranda-Filloy, J. A.; Blanco, Ricardo; Balsa, A.; Pascual-Salcedo, Dora; Rodríguez-Rodríguez, Luis; Fernández-Gutiérrez, B.; Ortiz, A M; González-Alvaro, I.; Gómez-Vaquero, C.; Bottini, N.; Llorca, Javier

    2011-01-01

    Abstract Introduction Acid phosphatase locus 1 (ACP1) encodes a low molecular weight phosphotyrosine phosphatase implicated in a number of different biological functions in the cell. The aim of this study was to determine the contribution of ACP1 polymorphisms to susceptibility to rheumatoid arthritis (RA), as well as the potential contribution of these polymorphisms to the increased risk of cardiovascular disease (CV) observed in RA patients. Methods A set of 1,603 Spanish RA patients and 1,...

  4. cDNA isolated from a human T-cell library encodes a member of the protein-tyrosine-phosphatase family

    A human peripheral T-cell cDNA library was screened with two labeled synthetic oligonucleotides encoding regions of a human placenta protein-tyrosine-phosphatase. One positive clone was isolated and the nucleotide sequence was determined. It contained 1,305 base pairs of open reading frame followed by a TAA stop codon and 978 base pairs of 3' untranslated end, although a poly(A)+ tail was not found. An initiator methionine residue was predicted at position 61, which would result in a protein of 415 amino acid residues. This was supported by the synthesis of a Mr 48,000 protein in an in vitro reticulocyte lysate translation system using RNA transcribed from the cloned cDNA and T7 RNA polymerase. The deduced amino acid sequence was compared to other known proteins revealing 65% identity to the low Mr PTPase 1B isolated from placenta. In view of the high degree of similarity, the T-cell cDNA likely encodes a newly discovered protein-tyrosine-phosphatase, thus expanding this family of genes

  5. Control of Acid Phosphatases Expression from Aspergillus niger by Soil Characteristics

    Ely Nahas

    2015-10-01

    Full Text Available ABSTRACTThis work studied the acid phosphatase (APase activity from culture medium (extracellular, eAPase and mycelial extract (intracellular, iAPase ofAspergillus niger F111. The influence of fungus growth and phosphate concentration of the media on the synthesis and secretion of phosphatase was demonstrated. The effects of pH, substrate concentration and inorganic and organic compounds added to the reaction mixture on APase activity were also studied. Both enzymes were repressed by high concentrations of phosphate. Overexpression of iAPase in relation to eAPase was detected; iAPase activity was 46.1 times higher than eAPase. The maximal activity of eAPase was after 24h of fungus growth and for iAPase was after 96h. Optimal pH and substrate concentrations were 4.5 and 8.0 mM, respectively. Michaelis-Menten constant (Km for the hydrolysis of p-nitrophenyl phosphate was 0.57 mM with Vmax = 14,285.71 U mg-1 mycelium for the iAPase and 0.31 mM with V max = 147.06 U mg-1 mycelium for eAPase. Organic substances had little effect on acid phosphatases when compared with the salts. Both the APases were inhibited by 10 mM KH 2PO4 and 5 mM (NH42MoO4; eAPase was also inhibited by 1 mM CoCl2.

  6. Purification and characterization of 29 kda acid phosphatase from germinating melon seeds

    Not much progress on the purification and characterization of low molecular weight acid phosphatases from plants has been made as yet. In the current study a low molecular weight acid phosphatase from seedling of melon was purified about 114-fold with specific activity of 45 U/ mg of protein and a recovery of 3 %. The enzyme was found to be homogeneous and showed a single band corresponding to 29 kDa on SDS-polyacrylamide gel electrophoresis. The Km for p-nitrophenyl phosphate was found to be 0.175 mM and Vmax was 42 micro mol of substrate hydrolyzed /min/mg of protein at pH 5.5 and at 37 degree C. The enzyme showed its optimum activity at pH 5.0 and 50 degree C. The enzyme was thermostable and it retained 70 % activity for 45 min at 60 degree C. The pH stability was 4.8-6.0. Phosphate, vanadate, molybdate and fluoride acted as strong inhibitors. Metal ions such as Zn /sup +2/, Cu /sup +2/, Ag /sup +2/ and Hg /sup +2/ deactivated the enzyme while other divalent ions such as Ca /sup +2/ and Mg /sup +2/ had no effect. (author)

  7. Use of acid phosphatase as biomarker during the castor bean seeds germination (ricinus communis

    Carmen Ferreira Veríssima

    2008-12-01

    Full Text Available One of the main oil crop of prominent social and economic importance is to mamoneira (Ricinus communis L.; with countless application in the industry and agricultural. Broadly it distributed in Brazil; his cultivation can be an alternative of sustainability in the Brazilian northeast. It know the physiological and biochemical mechanisms of the germination they are important for the best utilization of the plant. The objective of this work was use acid phosphatase as biomarker during the germination. In the rough extract occurred the dosage of the activity for pNPP; Tyr-Pi and PPi; determination of protein and inorganic phosphatse. The peak of activity for pNPP was in the seventh day; for PPi and Tyr-Pi in the ninth and for PEP in the fifth. The concentration of protein increased according to the days of germination; with peak of activity in the eighth day; being coincidental with the peaks of the activities for the substrates. The content of inorganic phosphate diminished with the time of germination and after the third day occurred a fall accentuated of its concentration. We concluded that acid phosphatase is important for the germination of the seeds and his paper is related with the mobilization of inorganic phosphate; the main nutrients for the development.

  8. 18O isotope effect in 13C nuclear magnetic resonance spectroscopy. Part 9. Hydrolysis of benzyl phosphate by phosphatase enzymes and in acidic aqueous solutions

    The 18O isotope-induced shifts in 13C and 31P nuclear magnetic resonance (NMR) spectroscopy were used to establish the position of bond cleavage in the phosphatase-catalyzed and acid-catalyzed hydrolysis reactions of benzyl phosphate. The application of the 18O-isotope effect in NMR spectroscopy affords a continuous, nondestructive assay method for following the kinetics and position of bond cleavage in the hydrolytic process. The technique provides advantages over most discontinuous methods in which the reaction components must be isolated and converted to volatile derivatives prior to analysis. In the present study, [α-13C,ester-18O]benzyl phosphate and [ester-18O]benzyl phosphate were synthesized for use in enzymatic and nonenzymatic studies. Hydrolysis reactions catalyzed by the alkaline phosphatase from E. coli and by the acid phosphatases isolated from human prostate and human liver were all accompanied by cleavage of the substrate phosphorus-oxygen bond consistent with previously postulated mechanisms involving covalent phosphoenzyme intermediates. An extensive study of the acid-catalyzed hydrolysis of benzyl phosphate at 750C revealed that the site of bond cleavage is dependent on pH. At pH less than or equal to 1.3, the hydrolysis proceeds with C-O bond cleavage; at 1.3 2H]Benzyl phosphate was also synthesized. Hydrolysis of this chiral benzyl derivative demonstrated that the acid-catalyzed C-O bond scission of benzyl phosphate proceeds by an A-1 (S/sub N/1) mechanism with 70% racemization and 30% inversion at carbon. 37 references, 4 figures, 2 tables

  9. An Approach to More Accurate Model Systems for Purple Acid Phosphatases (PAPs).

    Bernhardt, Paul V; Bosch, Simone; Comba, Peter; Gahan, Lawrence R; Hanson, Graeme R; Mereacre, Valeriu; Noble, Christopher J; Powell, Annie K; Schenk, Gerhard; Wadepohl, Hubert

    2015-08-01

    The active site of mammalian purple acid phosphatases (PAPs) have a dinuclear iron site in two accessible oxidation states (Fe(III)2 and Fe(III)Fe(II)), and the heterovalent is the active form, involved in the regulation of phosphate and phosphorylated metabolite levels in a wide range of organisms. Therefore, two sites with different coordination geometries to stabilize the heterovalent active form and, in addition, with hydrogen bond donors to enable the fixation of the substrate and release of the product, are believed to be required for catalytically competent model systems. Two ligands and their dinuclear iron complexes have been studied in detail. The solid-state structures and properties, studied by X-ray crystallography, magnetism, and Mössbauer spectroscopy, and the solution structural and electronic properties, investigated by mass spectrometry, electronic, nuclear magnetic resonance (NMR), electron paramagnetic resonance (EPR), and Mössbauer spectroscopies and electrochemistry, are discussed in detail in order to understand the structures and relative stabilities in solution. In particular, with one of the ligands, a heterovalent Fe(III)Fe(II) species has been produced by chemical oxidation of the Fe(II)2 precursor. The phosphatase reactivities of the complexes, in particular, also of the heterovalent complex, are reported. These studies include pH-dependent as well as substrate concentration dependent studies, leading to pH profiles, catalytic efficiencies and turnover numbers, and indicate that the heterovalent diiron complex discussed here is an accurate PAP model system. PMID:26196255

  10. Adsorption of Acid Phosphatase on Minerals and Soil Colloids in Presence of Citrate and Phosphate

    2002-01-01

    The aim of this work was to study the influence of phosphate and citrate, which are common inorganic andorganic anions in soils, on the adsorption of acid phosphatase by kaolin, goethite and the colloids separatedfrom yellow-brown soil (YBS) and latosol (LS) in central-south China. The YBS colloid has the major claymineral composition of 1.4 nm mineral, illite and kaolinite while the LS colloid mainly contains kaolinite andoxides. The adsorption isotherm of acid phosphatase on the examined soil colloids and minerals fitted tothe Langmuir model. The amount of enzyme adsorbed in the absence of ligands was in the order of YBScolloid >LS colloid>kaolin≈goethite. In the presence of phosphate or citrate, the amounts of the enzymeadsorbed followed the sequence YBS colloid>kaolin>LS colloid>goethite. The presence of ligands alsodecreased the binding energy between the enzyme and soil colloids or minerals. With the increase of ligandconcentration from 10 mmol L-1 to 400 m mol L-1, different behaviors for the adsorption of enzyme werefound in the colloid and mineral systems studied. A sharp decrease in enzyme adsorption was observed ongoethite while gradual decreases of enzyme adsorption were recorded in the two soil colloid systems. However,no any decrease was found for the amount of enzyme adsorbed on kaolin at higher ligand concentrations.When phosphate or citrate was introduced to the system before the addition of enzyme, the ligands usuallyenhanced the adsorption of enzyme. The results obtained in this study suggested the important role ofkaolinite mineral in the adsorption of enzyme molecules in acidic soils in the presence of various ligands.

  11. 130 kDa phosphatase from the liver of labeo rohita: isolation: purification and some kinetic properties

    An isoenzyme of high molecular weight acid phosphatase (HM-ACP) from the live of fish rohu (Labeo Rohita) was isolated and purified to homogeneity. The enzyme had specific activity of 14.96 U/mg and a recovery of about 4%. The purification procedure included ammonium sulphate precipitation and series of chromatographic separations on SP-Sephadex C-50, CM-Cellulose and Sephacryl HR-200 columns. Nealry 500-folds purification was achieved. The molecular weight was estimated to be 120-130 kDa by polyacrylamide gel electrophoresis (PAGE) of native enzyme and 130 kDa by gel filtration on calibrated Sephadex G-100 column. sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) under reduced and non-reduced condition showed a band corresponding to 66 kDa confirming the dimeric nature of enzyme. para nitrophenyl phosphate and flavin mononucleotide were hydrolyzed effectively by the enzyme and found to be good substrates. Optimum temperature for the enzyme was 50 degree C and temperature stability was 0 degree-50 degree C. Similarly optimum ph for the enzyme was 5.4 and ph stability was 4.8-6.0. The K/sub m/ for the p-nitrophenyl phosphate was estimated to be 0.15 mM. The enzyme was competitively inhibited by the phosphate, vanadate, molybdate, tartrate, fluoride and pyridoxal-5-PO/sub 4/ while pyridoxamine-5-PO/sub 4/ showed poor inhibition. Metal ions such as Ag/sup +/, Cu/sup ++/ Zn/sup ++/ showed strong inhibition on the enzyme activity while other divalent ions like Mg/sup ++/, Mn/sup ++/ and Co/sup ++/ were found to be poor inhibitors. Modifiers like EDTA, methanol, ethanol, acetone and glycerol had no effect on the enzyme's activity. (author)

  12. Localization of acid phosphatase activity in the apoplast of root nodules of pea (Pisum sativum

    Marzena Sujkowska

    2011-04-01

    Full Text Available Changes in the activity of acid phosphatase (AcPase in the apoplast of pea root nodule were investigated. The activity was determined using lead and cerium methods. The results indicated a following sequence of AcPase activity appearance during the development of the infection thread: 1 low AcPase activity appears in the outer part of cells of symbiotic bacteria; 2 bacteria show increased AcPase activity, and the enzyme activity appears in the thread walls; 3 activity exhibits also matrix of the infection thread; 4 bacteria just before their release from the infection threads show high AcPase activity; 5 AcPase activity ceases after bacteria transformation into bacteroids. The increase in bacterial AcPase activity may reflect a higher demand for inorganic phosphorus necessary for propagation of the bacteria within the infection threads and/or involved in bacteria release from the infection threads.

  13. Strigolactone regulates anthocyanin accumulation, acid phosphatases production and plant growth under low phosphate condition in Arabidopsis.

    Shinsaku Ito

    Full Text Available Phosphate is an essential macronutrient in plant growth and development; however, the concentration of inorganic phosphate (Pi in soil is often suboptimal for crop performance. Accordingly, plants have developed physiological strategies to adapt to low Pi availability. Here, we report that typical Pi starvation responses in Arabidopsis are partially dependent on the strigolactone (SL signaling pathway. SL treatment induced root hair elongation, anthocyanin accumulation, activation of acid phosphatase, and reduced plant weight, which are characteristic responses to phosphate starvation. Furthermore, the expression profile of SL-response genes correlated with the expression of genes induced by Pi starvation. These results suggest a potential overlap between SL signaling and Pi starvation signaling pathways in plants.

  14. Effect of copper on acid phosphatase activity in yeast Yarrowia lipolytica

    Ito, Hiroyasu; Inouhe, Masahiro; Tohoyama, Hiroshi; Joho, Masanori [Ehime Univ., Matsuyama (Japan). Dept. of Biology

    2007-01-15

    Acid phosphatase (APase) activity of the yeast Yarrowia lipolytica increased with increasing Cu{sup 2+} concentrations in the medium. Furthermore, the enzyme in soluble form was stimulated in vitro by Cu{sup 2+}, Co{sup 2+}, Ni{sup 2+}, Mn{sup 2+} and Mg{sup 2+} and inhibited by Ag{sup +} and Cd{sup 2+}. The most effective ion was Cu{sup 2+}, especially for the enzyme from cultures in medium containing Cu{sup 2+}, whereas APase activity in wall-bound fragments was only slightly activated by Cu{sup 2+}. The content of cellular phosphate involving polyphosphate was decreased by adding Cu{sup 2+}, regardless of whether or not the medium was rich in inorganic phosphate. Overproduction of the enzyme stimulated by Cu{sup 2+} might depend on derepression of the gene encoding the APase isozyme. (orig.)

  15. Optimization of the chloramine T method for radioiodination of prostate gland by acid phosphatase

    Optimal conditions for preparation of [125I]-labelled human prostatic acid phosphatase (PAP) by chloramin T method were developed which make it possible to increase the radiochemical yield and immunoreactivity of labelled PAP. The influence of buffer system, pH of reaction mixture and molarity of borate buffer was investigated. Iodination in 0.2 M borate buffer, pH 8.0 make it possible to prepare [125I]-PAP with 90-98 immunoreactivity. Under storage of labelled PAP during 2 month immunoreactivity is decreased only by 10%. On the base of prepared 125I]-PAP high-sensitive competitive radioimmunoassay of PAP in human serum have developed

  16. Radioimmunological and enzymatic assay for prostatic acid phosphatase (PAP) in prostatic cancer

    Serum prostatic acid phosphatase (PAP) level was determined by radioimmunoassay (RIA) in 272 patients. For comparative purposes PAP was also measured by an enzymatic assay. In prostate adenocarcinoma 55% of the values were elevated. In early stages (A and B) 17% of patients were found to be positive; at later stages (C and D) the percentage increased to 78%. The enzymatic method yielded 46% positive values in these patients: 17% in the former group (A+B), and 64% in the latter one (C+D). False positive values were observed in 10% of the patients with prostate adenoma, and 22% of the patients with prostatis. The data confirm the low sensitivity of RIA (and enzymatic assay) for detecting early intracapsular disease. RIA determination of PAP is a good diagnostic tool for advanced cancer of the prostate. (author)

  17. Histochemical study on effects of low dose γ-irradiation on acid phosphatase in liver of the pigeons Columbus livia intermedia Strickland

    Effects of total body γ-irradiation with sub-lethal dose (400 rads) on acid phosphatase have been studied in the liver of pigeon. The histochemical study showed increased activity of acid phosphatase in liver after 48 hr and 72 hr of irradiation. (author)

  18. Specific activity of cell-surface acid phosphatase in different bacterioplankton morphotypes in an acidified mountain lake

    Nedoma, Jiří; Vrba, Jaroslav

    2006-01-01

    Roč. 8, č. 7 (2006), s. 1271-1279. ISSN 1462-2912 R&D Projects: GA AV ČR(CZ) IAA6017202 Institutional research plan: CEZ:AV0Z60170517 Keywords : alkaline phosphatase * bacterial morphorypes * acid ified lake Subject RIV: CE - Biochemistry Impact factor: 4.630, year: 2006

  19. The Leishmania donovani histidine acid ecto-phosphatase LdMAcP: insight into its structure and function.

    Papadaki, Amalia; Politou, Anastasia S; Smirlis, Despina; Kotini, Maria P; Kourou, Konstadina; Papamarcaki, Thomais; Boleti, Haralabia

    2015-05-01

    Acid ecto-phosphatase activity has been implicated in Leishmania donovani promastigote virulence. In the present study, we report data contributing to the molecular/structural and functional characterization of the L. donovani LdMAcP (L. donovani membrane acid phosphatase), member of the histidine acid phosphatase (HAcP) family. LdMAcP is membrane-anchored and shares high sequence identity with the major secreted L. donovani acid phosphatases (LdSAcPs). Sequence comparison of the LdMAcP orthologues in Leishmania sp. revealed strain polymorphism and species specificity for the L. donovani complex, responsible for visceral leishmaniasis (Khala azar), proposing thus a potential value of LdMAcP as an epidemiological or diagnostic tool. The extracellular orientation of the LdMAcP catalytic domain was confirmed in L. donovani promastigotes, wild-type (wt) and transgenic overexpressing a recombinant LdMAcP-mRFP1 (monomeric RFP1) chimera, as well as in transiently transfected mammalian cells expressing rLdMAcP-His. For the first time it is demonstrated in the present study that LdMAcP confers tartrate resistant acid ecto-phosphatase activity in live L. donovani promastigotes. The latter confirmed the long sought molecular identity of at least one enzyme contributing to this activity. Interestingly, the L. donovani rLdMAcP-mRFP1 promastigotes generated in this study, showed significantly higher infectivity and virulence indexes than control parasites in the infection of J774 mouse macrophages highlighting thereby a role for LdMAcP in the parasite's virulence. PMID:25695743

  20. Optimal level of Purple Acid Phosphatase5 is required for maintaining complete resistance to Pseudomonas syringae

    Sridhar eRavichandran

    2015-08-01

    Full Text Available Plants possess an exceedingly complex innate immune system to defend against most pathogens. However, a relative proportion of the pathogens overcome host’s innate immunity and impair plant growth and productivity. We previously showed that mutation in purple acid phosphatase (PAP5 lead to enhanced susceptibility of Arabidopsis to the bacterial pathogen Pseudomonas syringae pv tomato DC3000 (Pst DC3000. Here, we report that an optimal level of PAP5 is crucial for mounting complete basal resistance. Overexpression of PAP5 impaired ICS1, PR1 expression and salicylic acid (SA accumulation similar to pap5 knockout mutant plants. Moreover, plant overexpressing PAP5 was impaired in H2O2 accumulation in response to Pst DC3000. PAP5 is localized in to peroxisomes, a known site of generation of reactive oxygen species for activation of defense responses. Taken together, our results demonstrate that optimal levels of PAP5 is required for mounting resistance against Pst DC3000 as both knockout and overexpression of PAP5 lead to compromised basal resistance.

  1. Tunable phosphatase-sensitive stable prodrugs of 5-aminolevulinic acid for tumor fluorescence photodetection.

    Babič, Andrej; Herceg, Viktorija; Ateb, Imène; Allémann, Eric; Lange, Norbert

    2016-08-10

    5-Aminolevulinic acid (5-ALA) has been at the forefront of small molecule based fluorescence-guided tumor resection and photodynamic therapy. 5-ALA and two of its esters received marketing authorization but suffer from several major limitations, namely low stability and poor pharmacokinetic profile. Here, we present a new class of 5-ALA derivatives aiming at the stabilization of 5-ALA by incorporating a phosphatase sensitive group, with or without self-cleavable linker. Compared to 5-ALA hexyl ester (5-ALA-Hex), these compounds display an excellent stability under acidic, basic and physiological conditions. The activation and conversion into the 5-ALA is controlled and can be structure-tailored. The prodrugs display reduced acute toxicity compared to 5-ALA-Hex with superior dose response profiles of protoporphyrin IX synthesis and fluorescence intensity in human glioblastoma cells in vitro. Clinically relevant fluorescence kinetics in vivo shown in U87MG glioblastoma spheroid tumor model in chick embryos provide a solid basis for their further development and translation to clinical fluorescence guided tumor resection and photodynamic therapy. PMID:27235981

  2. Overexpression of OsPAP10a, A Root-Associated Acid Phosphatase, Increased Extracellular Organic Phosphorus Utilization in Rice

    Jingluan Tian; Chuang Wang; Qian Zhang; Xiaowei He; James Whelan; Huixia Shou

    2012-01-01

    Phosphorus (P) deficiency is a major limitation for plant growth and development.Among the wide set of responses to cope with low soil P,plants increase their level of intracellular and secreted acid phosphatases (APases),which helps to catalyze inorganic phosphate (Pi) hydrolysis from organophosphates.In this study we characterized the rice (Oryza sativa) purple acid phosphatase 10a (OsPAP10a).OsPAP10a belongs to group la of purple acid phosphatases (PAPs),and clusters with the principal secreted PAPs in a variety of plant species including Arabidopsis.The transcript abundance of OsPAP10a is specifically induced by Pi deficiency and is controlled by OsPHR2,the central transcription factor controlling Pi homeostasis.In gel activity assays of root and shoot protein extracts,it was revealed that OsPAP10a is a major acid phosphatase isoform induced by Pi starvation.Constitutive overexpression of OsPAP10a results in a significant increase of phosphatase activity in both shoot and root protein extracts.In vivo root 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) assays and activity measurements on external media showed that OsPAP10a is a root-associated APase.Furthermore,overexpression of OsPAP10a significantly improved ATP hydrolysis and utilization compared with wild type plants.These results indicate that OsPAP10a can potentially be used for crop breeding to improve the efficiency of P use.

  3. Ser/Thr-rich repetitive motifs as targets for phosphoglycan modifications in Leishmania mexicana secreted acid phosphatase.

    Wiese, M; Ilg, T; Lottspeich, F; Overath, P

    1995-03-15

    The insect stage of the protozoan parasite Leishmania mexicana secretes a phosphomonoesterase in the form of a filamentous complex. The polypeptide subunits of this polymer are modified by phosphoglycans and/or oligomannosyl residues linked to phosphoserine. Based on peptide sequence data of a predominant 100 kDa protein of the filamentous complex, two tandemly arranged, single copy genes, lmsap1 and lmsap2, were cloned and sequenced. lmsap1 predicts a protein with features characteristic of acid phosphatases and a remarkable serine- and threonine-rich region of 32 amino acids close to the C-terminus. In the otherwise identical lmsap2 product, this region is extended to 383 amino acids and is composed of short Ser/Thr-rich repeats. Deletion analysis demonstrates that lmsap1 encodes the major 100 kDa protein of the complex while a minor 200 kDa component is derived from the lmsap2 gene. Null mutants of either gene retain the ability to secrete acid phosphatase filaments, while a deletion of both genes results in Leishmania defective in enzyme formation. The Ser/Thr-rich domains are the targets for phosphoglycan modifications as shown by the expression of secreted fusion proteins composed of these C-terminal regions and the N-terminal domain of a lysosomal acid phosphatase. PMID:7720697

  4. Phosphatase inhibitors remove the run-down of γ-aminobutyric acid type A receptors in the human epileptic brain

    Palma, E.; Ragozzino, D. A.; Di Angelantonio, S.; Spinelli, G.; Trettel, F.; Martinez-Torres, A.; Torchia, G.; Arcella, A.; Di Gennaro, G.; Quarato, P. P.; Esposito, V.; Cantore, G.; Miledi, R.; Eusebi, F.

    2004-01-01

    The properties of γ-aminobutyric acid (GABA) type A receptors (GABAA receptors) microtransplanted from the human epileptic brain to the plasma membrane of Xenopus oocytes were compared with those recorded directly from neurons, or glial cells, in human brains slices. Cell membranes isolated from brain specimens, surgically obtained from six patients afflicted with drug-resistant temporal lobe epilepsy (TLE) were injected into frog oocytes. Within a few hours, these oocytes acquired GABAA receptors that generated GABA currents with an unusual run-down, which was inhibited by orthovanadate and okadaic acid. In contrast, receptors derived from membranes of a nonepileptic hippocampal uncus, membranes from mouse brain, or recombinant rat α1β2γ2-GABA receptors exhibited a much less pronounced GABA-current run-down. Moreover, the GABAA receptors of pyramidal neurons in temporal neocortex slices from the same six epileptic patients exhibited a stronger run-down than the receptors of rat pyramidal neurons. Interestingly, the GABAA receptors of neighboring glial cells remained substantially stable after repetitive activation. Therefore, the excessive GABA-current run-down observed in the membrane-injected oocytes recapitulates essentially what occurs in neurons, rather than in glial cells. Quantitative RT-PCR analyses from the same TLE neocortex specimens revealed that GABAA-receptor β1, β2, β3, and γ2 subunit mRNAs were significantly overexpressed (8- to 33-fold) compared with control autopsy tissues. Our results suggest that an abnormal GABA-receptor subunit transcription in the TLE brain leads to the expression of run-down-enhanced GABAA receptors. Blockage of phosphatases stabilizes the TLE GABAA receptors and strengthens GABAergic inhibition. It may be that this process can be targeted to develop new treatments for intractable epilepsy. PMID:15218107

  5. Mice deficient in transmembrane prostatic acid phosphatase display increased GABAergic transmission and neurological alterations.

    Heidi O Nousiainen

    Full Text Available Prostatic acid phosphatase (PAP, the first diagnostic marker and present therapeutic target for prostate cancer, modulates nociception at the dorsal root ganglia (DRG, but its function in the central nervous system has remained unknown. We studied expression and function of TMPAP (the transmembrane isoform of PAP in the brain by utilizing mice deficient in TMPAP (PAP-/- mice. Here we report that TMPAP is expressed in a subpopulation of cerebral GABAergic neurons, and mice deficient in TMPAP show multiple behavioral and neurochemical features linked to hyperdopaminergic dysregulation and altered GABAergic transmission. In addition to increased anxiety, disturbed prepulse inhibition, increased synthesis of striatal dopamine, and augmented response to amphetamine, PAP-deficient mice have enlarged lateral ventricles, reduced diazepam-induced loss of righting reflex, and increased GABAergic tone in the hippocampus. TMPAP in the mouse brain is localized presynaptically, and colocalized with SNARE-associated protein snapin, a protein involved in synaptic vesicle docking and fusion, and PAP-deficient mice display altered subcellular distribution of snapin. We have previously shown TMPAP to reside in prostatic exosomes and we propose that TMPAP is involved in the control of GABAergic tone in the brain also through exocytosis, and that PAP deficiency produces a distinct neurological phenotype.

  6. Comparative theoretical studies of the phosphomonoester hydrolysis mechanism by purple acid phosphatases.

    Retegan, M; Milet, A; Jamet, H

    2010-07-01

    We present here the first ONIOM (our own n-layered integrated molecular orbital + molecular mechanics method) studies of a purple acid phosphatase enzyme. Our study focused on the structures of the red kidney bean PAP (kbPAP) complexed with phosphate and with phenyl phosphate and on the mechanism of the phenyl phosphate hydrolysis by the enzyme. Density functional theory (DFT) calculations were also performed using models of different sizes for comparison purpose. Results show that the inclusion of three histidine residues, His202, His295, and His296, with their protein surrounding, is crucial to properly describe the coordination of the substrates. They induce a conformation with the substrate closer to the nucleophilic mu-hydroxyde bridge. In the mechanistic study, a transition state is stabilized by a strong hydrogen bond between His202 and the leaving group of the substrate. Consequently, a smaller value for the activation energy barrier is obtained from DFT calculations including this histidine to the same calculations without this histidine. Using the ONIOM method, this activation energy barrier is even more reduced. So the mechanism, which considers the hydroxo group bridging the two metal ions as nucleophile, becomes really convincing, contrary to the results obtained with a small model at the DFT level. PMID:20550096

  7. Cloning and Characterization of Purple Acid Phosphatase Phytases from Wheat, Barley, Maize and Rice

    Dionisio, Giuseppe; Madsen, Claus Krogh; Holm, Preben Bach;

    2011-01-01

    is demonstrated that wheat, barley, maize, and rice all possess purple acid phosphatase (PAP) genes that, expressed in Pichia pastoris, give fully functional phytases (PAPhys) with very similar enzyme kinetics. Preformed wheat PAPhy was localized to the protein crystalloid of the aleurone vacuole......Barley (Hordeum vulgare) and wheat (Triticum aestivum) possess significant phytase activity in the mature grains. Maize (Zea mays) and rice (Oryza sativa) possess little or virtually no preformed phytase activity in the mature grain and depend fully on de novo synthesis during germination. Here, it....... Phylogenetic analyses indicated that PAPhys possess four conserved domains unique to the PAPhys. In barley and wheat, the PAPhy genes can be grouped as PAPhy_a or PAPhy_b isogenes (barley, HvPAPhy_a, HvPAPhy_b1, and HvPAPhy_b2; wheat, TaPAPhy_a1, TaPAPhy_a2, TaPAPhy_b1, and TaPAPhy_b2). In rice and maize, only...

  8. Fundamental studies of three radioimmunoassay kits measuring prostatic acid phosphatase (PAP) concentrations

    Measurement of prostatic acid phosphatase (PAP) concentration is useful for the diagnosis and treatment of prostatic cancer. Fundamental studies of measurement of PAP concentration by radioimmunoassay were performed and values determined by three commercially available kits were compared, those are, PAP EIKEN RIA Kit (E kit), RIA Quant P.A.P. (M kit) and GammaDab PAP RIA Kit (C kit). Upper limits of normal PAP concentration were 3 ng/ml by E and M kits and 2 ng/ml by C kits, respectively. The reproducibility and the recovery studies of three kits were satisfactory. However, dilution curve of some patients was not strait. PAP concentration of 27 patients measured by three kits were correlated well to each other, but the discrepancy of values was noticed in high PAP concentrations. PAP concentration measured by RIA is more reliable than that by enzyme immunoassay. When keeping serum samples, the effect of time, temperature and freeze and thawing on PAP values was obvious. It is recommended that serum is separated at 40C as soon as possible after collecting blood and kept frozen until use. (author)

  9. Studies on alkaline and acid phosphatase activity of neutrophil leukicytes, 2

    With a view to analyzing the inhibiting effect of anticancer drugs and irradiation on hematopoiesis in rabbits neutrophil (pseudoeosinophil) counts and the neutrophilic activities of alkaline phosphatase (AP) and acid phosphatase (SP) were serially followed up after drug administration or irradiation. The enzym activity was estimated histochemically, using azo-dye staining. Each rabbit was given cyclophosphamid (CP) (25mg/kg x 10, at intervals of 5 - 7 days ; 50mg/kg x 5, every day; or 100mg/kg x 1, i.m.), Thio-TEPA (4mg/kg x 1, i.m.), Vinblastin (VBT) (1mg/kg x 1, i.v.), 6MP (25mg/kg x 1, p.o.), or Mitomycin C (MMC) (1.5mg/kg x 1, i.v.). The results obtained were as follows : 1) The neutrophil counts became slightly elevated at 24 hrs, reached their nadir at 48 to 72 hrs, and recovered to normal in 5 to 6 days thereafter, except with 6 MP which produced no significant change but for a temporary elevation after dosages. 2) Except in the group administrated 6MP, which caused no significant hematorogical changes, the AP changes were similar in all of the animal groups : after temporary depression, it became elevated for 5 to 6 days, and recovered to normal about 9 days thereafter. 3) SP showed no changes in the 25mg/kg x 10 CP and the 6MP groups, it became elevated in 2 or 3 days after the administration of MMC, VBT, or Thio-TEPA to recover to normal in 5 to 10 days thereafter. 4) 60Co irradiation (1,000 rad/whole body x 1) led to a temporary ascent in phil count followed by a descent from the 6th day on, and then a slow recovery to normal. AP was elevated from the third to the sixth days, and, after a depression on the tenth day, it returned to normal 24 days after irradiation, while SP showed a continued elevation from the 2nd to the 13th day. (author)

  10. Evaluation of the Staphylococcus aureus Class C Nonspecific Acid Phosphatase (SapS) as a Reporter for Gene Expression and Protein Secretion in Gram-Negative and Gram-Positive Bacteria▿

    Du Plessis, Erika; Theron, Jacques; Berger, Eldie; Louw, Maureen

    2007-01-01

    A phosphatase secreted by Staphylococcus aureus strain 154 has previously been characterized and classified as a new member of the bacterial class C family of nonspecific acid phosphatases. As the acid phosphatase activity can be easily detected with a cost-effective plate screen assay, quantitatively measured by a simple enzyme assay, and detected by zymography, its potential use as a reporter system was investigated. The S. aureus acid phosphatase (sapS) gene has been cloned and expressed f...

  11. Study of possible changes brought about by plutonium oxide in the acid phosphatase activity of alveolar macrophages of the rabbit

    This report describes the various techniques used for determining the acid phosphatase activity of alveolar rabbit macrophages after inhalation of radioactive plutonium oxide particles, exposure of the animals, removal and sampling of the alveolar cells, and technical dosage. The results obtained are presented; they do not make it possible, in this particular case, to affirm that an important change in the enzymatic activity studied occurs. (author)

  12. Tartrate resistant acid phosphatase 5a : a potential regulator of adipocyte cell number and differentiation in white adipose tissue

    Patlaka, Christina

    2015-01-01

    Tartrate- resistant acid phosphatase (TRAP) exists in two isoforms, TRAP 5a which is monomeric and TRAP 5b which is a dimer generated by proteolytic cleavage of TRAP 5a, that exhibit different functions and localizations. TRAP 5a is expressed by adipose tissue macrophages and secreted into the extracellular environment and has been shown to lead to hyperplastic insulin- sensitive obesity when over-expressed in mice. In bone, TRAP is suggested to interact with the heparan sulfat...

  13. The Effects of Two Species of Daphne, Betulin and Betulinic Acid on Alkaline Phosphatase Activity in Two Human Cancer Cell lines, K562 and MCF-7

    E Panahi Kokhdan

    2014-02-01

    Full Text Available Abstract Background & aim: Changes of alkaline phosphatase activity is one of the symptoms of many diseases. The aim of this study was to evaluate the effect of two types of Daphne, Betulin and Betulinic acid, on alkaline phosphatase activity in K562 and MCF-7 cell lines, respectively. Methods: In this study, 106 cancer cell lines of K562 and MCF-7 were cultured in presence of 5% carbon dioxide at 37 ° C. at doses near the IC50. The viability of cells, inside and outside alkaline phosphatase activity and the amount of total protein in each treatment were studied. The collected data was analyzed with a multivariate analysis of variance (Nested Design and Dunnett test. Results: The intracellular alkaline phosphatase activity of the cells showed different behavior compared to the extracellular alkaline phosphatase activity (p< 0.01. The highest increase of alkaline phosphatase activity in two cell lines (K562 and MCF-7 were 339% and 236% which was related to the treatment by macronata daphne. Conclusion: Unexpected increase in intracellular alkaline phosphatase activity in D. mucronata, D. oleides, Betulin, and Betulinic acid treatment may be due to changes in the composition of plasma membrane component and an increase the non-connected membrane of the protein which is due to the creation of more active proteins. Keywords: Daphne mucronata, Daphne oleoides, Alkaline Phosphatase, Betulinic Acid, Betulin

  14. ANTIMICROBIAL ACTIVITY OF LACTIC ACID BACTERIAL ISOLATES

    Utkarsha S. Shivsharan

    2013-08-01

    Full Text Available Micro-organisms have tendency to produce antimicrobial substances which show biological activity against other kind of micro-organisms. This phenomenon of bacterial antagonism is observed in lactic acid bacteria with competitive advantages. The lactic acid bacteria are commonly present in many fermented products, fruits and milk products. The variety of antimicrobial substances produced by lactic acid bacteria showing good inhibition capacity include production of lactic acid, acetic acid, hydrogen peroxide, carbon dioxide, diacetyl and bacteriocin. Bacteriocins produced by lactic acid bacteria are the subject of intense research because of their antimicrobial activity against food born bacteria such as Listeria monocytogenes, staphylococcus aureus, Bacillus cereus, Clostridium botulinum and several others .Bacteriocins may be bacteriostatic or bactericidal with narrow or broad range of activity. The main of the study was to study the antimicrobial activity of such lactic acid bacterial isolates.

  15. Isolation of humic acids from leonardite

    Shah, S.B.; Tartamella, T.L.; Lee, S. [Univ. of Akron, OH (United States). Dept. of Chemical Engineering; Kulik, C.J. [Ancon International, Newark, CA (United States)

    1996-12-31

    The primary interest in humic acid is its use as an effective fertilizer. Humic substances, found commonly in low-rank coals, enhance plant growth directly through positive physiological effects and indirectly by affecting the properties of the soil. Humic acids have traditionally been defined as the dark-colored organic matter that can be extracted from soil by dilute alkali and other reagents and which is insoluble in dilute acid. This paper discusses the isolation of humic acid from leonardite using the alkaline extraction method and the subsequent characterization using elemental analysis and infrared spectroscopy techniques. In this study, yields of more than 60% were obtained.

  16. Purification of a tartrate-resistant acid phosphatase (TrACP) from bovine cortical bone matrix

    It has been previously demonstrated that a partially purified bovine skeletal TrACP showed protein phosphatase (P'ase) activity that was specific for phosphotyrosyl (Ptyr) proteins. They have now purified TrACP activity from bovine cortical bone matrix to apparent homogeneity. The purification procedures included CM-Sepharose ion-exchange, cellulose phosphate affinity, sephacryl S-300 gel filtration and phenyl sepharose affinity chromatographies. Overall yield was > 25% and purification was approximately 2000-fold with a specific activity of 8.15 umol pNPP hydrolyzed/min/mg protein at 370C. The purified enzyme was judged to be homogeneous based on: (i) appearance as a single protein band on SDS-PAGE (silver staining technique) and (ii) distribution analysis of radioiodinated purified TrACP after SDS-PAGE revealing one band of radioactivity at the same positions as the TrACP protein band. M.W. of TrACP was 34,600 as assessed by gel filtration and 32,500 by SDS-PAGE, suggesting that bovine skeletal TrACP exists as active monomer. Analysis of the purified TrACP by isoelectric focusing showed at least 9 bands of enzyme activities with pIs between 4 and 5, indicating micro-heterogenecity. Substrate specificity analyses revealed that the purified TrACP also hydrolyzed nucleotide tri- and di-phosphates, but not monophosphates or other low M.W. phosphoryl esters, and was also capable of hydrolyzing phosphotyrosine (Tyr(P)) and Ptyr proteins with little activity toward other phosphoamino acids or phosphoseryl proteins. Optimal pH was 5.5 for TrACP activity, 6.0 for Tyr(P) P'ase activity and 7.0 for Ptyr protein P'ase activity. Results of these studies represent the first purification of a skeletal TrACP to apparent homogeneity

  17. Phosphate status and acid phosphatase activity in soil and ectomycorrhizas in two mature stands of scots pine (Pinus sylvestris L.) exposed to different levels of anthropogenic pollution

    Barbara Kieliszewska-Rokicka

    2014-01-01

    The relations between anthropogenic environmental pollution and the level of inorganic phosphorus in soil, enzyme activities of extracellular soil acid phosphatase and the surface acid phosphatase of excised ectomycorrhizas of Scots pine (Pinus sylvestris L.) were studied. Soil and root samples were taken from two Scots pine stands in central Poland: a polluted site exposed to long-term pollution from a steelworks and the city of Warsaw and a reference plot (control) free from direct impact o...

  18. Monoclonal antibodies directed against Leishmania secreted acid phosphatase and lipophosphoglycan. Partial characterization of private and public epitopes.

    Ilg, T; Harbecke, D; Wiese, M; Overath, P

    1993-10-15

    Leishmania promastigotes, the stage of the parasite characteristic for the sandfly vector, express an abundant glycoconjugate, called lipophosphoglycan, at their surface. Lipophosphoglycan consists of lysoalkyl-sn-glycerophosphoinositol linked to a phosphosaccharide core conserved in all species, which is connected to PO4-6Gal beta 1,4Man alpha 1 repeats with species-specific substitutions at the Gal residue; the repeats are capped by conserved and species-specific oligosaccharides. Most Leishmania species also secrete an acid phosphatase, which, in Leishmania mexicana, is a filamentous complex composed of a phosphorylated glycoprotein and non-covalently associated proteo-(high-molecular-mass)phosphoglycan. The secreted acid phosphatase complex was used as an antigen to derive a panel of monoclonal antibodies (mAbs). A total of 25 mAbs (17 novel and 8 previously described) were tested by different techniques for their specificity against lipophosphoglycan and secreted acid phosphatase from several Leishmania species. This comparison and the modification of the antigens by chemical or enzymic treatments allowed a classification of the mAbs into several groups. First, from 25 mAbs examined, 22 recognize lipophosphoglycan and the enzyme complex of L. mexicana; only three are specific for secreted acid phosphatase. Two of the latter group are also directed against carbohydrate structures, whereas the third mAb recognizes the 100-kDa polypeptide of the complex. The secreted acid-phosphatase-specific class detects antigen in the flagellar pocket of promastigotes while all anti-lipophosphoglycan mAbs bind to the cell surface. Second, all 15 anti-lipophosphoglycan mAbs investigated in detail appear to be directed against the phosphosaccharide repeats or the cap structure rather than the phosphosaccharide core. Two mAbs recognize terminal cap-structures containing Man alpha 1,2Man residues. Four antibodies are specific for L. mexicana and are probably directed against PO4

  19. Identification of genes required for secretion of the Francisella oxidative burst-inhibiting acid phosphatase AcpA

    John S Gunn

    2016-04-01

    Full Text Available Francisella tularensis is a Tier 1 bioterror threat and the intracellular pathogen responsible for tularemia in humans and animals. Upon entry into the host, Francisella uses multiple mechanisms to evade killing. Our previous studies have shown that after entering its primary cellular host, the macrophage, Francisella immediately suppresses the oxidative burst by secreting a series of acid phosphatases including AcpA-B-C and HapA, thereby evading the innate immune response of the macrophage and enhancing survival and further infection. However, the mechanism of acid phosphatase secretion by Francisella is still unknown. In this study, we screened for genes required for AcpA secretion in Francisella. We initially demonstrated that the known secretion systems, the putative Francisella-pathogenicity island (FPI-encoded Type VI secretion system and the Type IV pili, do not secrete AcpA. Using random transposon mutagenesis in conjunction with ELISA, Western blotting and acid phosphatase enzymatic assays, a transposon library of 5450 mutants was screened for strains with a minimum 1.5-fold decrease in secreted (culture supernatant AcpA, but no defect in cytosolic AcpA. Three mutants with decreased supernatant AcpA were identified. The transposon insertion sites of these mutants were revealed by direct genomic sequencing or inverse-PCR and sequencing. One of these mutants has a severe defect in AcpA secretion (at least 85% decrease and is a predicted hypothetical inner membrane protein. Interestingly, this mutant also affected the secretion of the FPI-encoded protein, VgrG. Thus, this screen identified novel protein secretion factors involved in the subversion of host defenses.

  20. Identification of Genes Required for Secretion of the Francisella Oxidative Burst-Inhibiting Acid Phosphatase AcpA.

    Hoang, Ky Van; Chen, Carolyn G; Koopman, Jacob; Moshiri, Jasmine; Adcox, Haley E; Gunn, John S

    2016-01-01

    Francisella tularensis is a Tier 1 bioterror threat and the intracellular pathogen responsible for tularemia in humans and animals. Upon entry into the host, Francisella uses multiple mechanisms to evade killing. Our previous studies have shown that after entering its primary cellular host, the macrophage, Francisella immediately suppresses the oxidative burst by secreting a series of acid phosphatases including AcpA-B-C and HapA, thereby evading the innate immune response of the macrophage and enhancing survival and further infection. However, the mechanism of acid phosphatase secretion by Francisella is still unknown. In this study, we screened for genes required for AcpA secretion in Francisella. We initially demonstrated that the known secretion systems, the putative Francisella-pathogenicity island (FPI)-encoded Type VI secretion system and the Type IV pili, do not secrete AcpA. Using random transposon mutagenesis in conjunction with ELISA, Western blotting and acid phosphatase enzymatic assays, a transposon library of 5450 mutants was screened for strains with a minimum 1.5-fold decrease in secreted (culture supernatant) AcpA, but no defect in cytosolic AcpA. Three mutants with decreased supernatant AcpA were identified. The transposon insertion sites of these mutants were revealed by direct genomic sequencing or inverse-PCR and sequencing. One of these mutants has a severe defect in AcpA secretion (at least 85% decrease) and is a predicted hypothetical inner membrane protein. Interestingly, this mutant also affected the secretion of the FPI-encoded protein, VgrG. Thus, this screen identified novel protein secretion factors involved in the subversion of host defenses. PMID:27199935

  1. Castor oil increases intestinal formation of platelet-activating factor and acid phosphatase release in the rat.

    Pinto, A; Calignano, A; Mascolo, N; Autore, G; Capasso, F

    1989-01-01

    1. When castor oil was administered by gavage to rats, the duodenum and jejunum but not ileum and colon produced large amounts (5-6 fold greater than control) of platelet activating factor (Paf). 2. Intraluminal release of acid phosphatase (AP) was also markedly increased (5-6 fold greater than control) in the duodenum and jejunum of castor oil-treated rats and there was a correlation between the elevated release of AP and intestinal hyperaemia. 3. These findings support a role for Paf as a m...

  2. Improving phosphorus acquisition of white clover (Trifolium repens L.) by transgenic expression of plant-derived phytase and acid phosphatase genes.

    Ma, Xue-Feng; Wright, Elane; Ge, Yaxin; Bell, Jeremey; Xi, Yajun; Bouton, Joseph H; Wang, Zeng-Yu

    2009-04-01

    Phosphate is one of the least available macronutrients restricting crop production in many ecosystems. A phytase gene (MtPHY1) and a purple acid phosphatase gene (MtPAP1), both isolated from the model legume Medicago truncatula, were introduced into white clover (Trifolium repens L.) by Agrobacterium-mediated transformation. The transgenes were driven by the constitutive CaMV35S promoter or the root-specific MtPT1 promoter. Transcripts were detected in roots of the transgenic plants. Phytase or acid phosphatase (APase) activities in root apoplasts of the transgenic plants were increased up to three-fold compared to the wild type control. After the plants were grown 80 days in sand pots supplied with organic phosphorus (Po) as the sole P source, dry weights of shoot tissues of the best performing transgenic plants almost doubled that of the control and were comparable to the counterparts supplied with inorganic phosphorus (Pi). Relative biomass production of the transgenics under Po treatment was over 90% and 80% of that from the Pi treatment when the plants were grown in hydroponics (40 days) and sand pots (80 days), respectively. In contrast, biomass of the wild type controls under Po treatment was only about 50% of the Pi treatment in either hydroponic cultures or sand pots. In addition, shoot P concentrations of the transgenic plants were significantly increased compared to the control. Transgenic plants accumulated much higher amounts of total P (up to 2.6-fold after 80 days of growth) than the control in Po supplied sand pots. The results showed that transgenic expression of MtPHY1 or MtPAP1 in white clover plants increased their abilities of utilizing organic phosphorus in response to P deficiency. PMID:26493137

  3. Phosphate status and acid phosphatase activity in soil and ectomycorrhizas in two mature stands of scots pine (Pinus sylvestris L. exposed to different levels of anthropogenic pollution

    Barbara Kieliszewska-Rokicka

    2014-02-01

    Full Text Available The relations between anthropogenic environmental pollution and the level of inorganic phosphorus in soil, enzyme activities of extracellular soil acid phosphatase and the surface acid phosphatase of excised ectomycorrhizas of Scots pine (Pinus sylvestris L. were studied. Soil and root samples were taken from two Scots pine stands in central Poland: a polluted site exposed to long-term pollution from a steelworks and the city of Warsaw and a reference plot (control free from direct impact of pollution. The polluted site was characterised by high concentration of trace elements (Cd, Pb, Cu, Zn, Mn, Cr and low level of inorganic phosphate in soil. This site had significantly lower enzyme activities of soil acid phosphatase (0.54 µmoles p-nitrophenol released g-1 dry weight h-1 and surface acid phosphatase of pine ectomycorrhizas (3.37 µmoles p-nitrophenol released g-1 fresh weight h-1 than the control site (1.36 µmoles p-nitrophenol released g-1 dry weight h-1 and 12.46 µmoles p-nitrophenol released g-1 fresh weight h-1, respectively. The levels of phosphate, carbon and nitrogen in pine fine roots were also analysed. Low concentrations of P04-P and high N: P ratio in pine fine roots from polluted site were found. The results suggest that soil pollutants may have a negative effect on the extracellular acid phosphatase of soil and Scots pine ectomycorrhizas and on the phosphorus status in fine roots of the plant.

  4. Uranium Biomineralization As a Result of Bacterial Phosphatase Activity: Insights From Bacterial Isolates From a Contaminated Subsurface

    Uranium contamination is an environmental concern at the Department of Energy's Field Research Center in Oak Ridge, Tennessee. In this study, we investigated whether phosphate biomineralization, or the aerobic precipitation of U(VI)-phosphate phases facilitated by the enzymatic activities of microorganisms, offers an alternative to the more extensively studied anaerobic U(VI) bioreduction. Three heterotrophic bacteria isolated from FRC soils were studied for their ability to grow and liberate phosphate in the presence of U(VI) and an organophosphate between pH 4.5 and 7.0. The objectives were to determine whether the strains hydrolyzed sufficient phosphate to precipitate uranium, to determine whether low pH might have an effect on U(VI) precipitation, and to identify the uranium solid phase formed during biomineralization. Two bacterial strains hydrolyzed sufficient organophosphate to precipitate 73-95% total uranium after 120 h of incubation in simulated groundwater. The highest rates of uranium precipitation and phosphatase activity were observed between pH 5.0 and 7.0. EXAFS spectra identified the uranyl phosphate precipitate as an autunite/meta-autunite group mineral. The results of this study indicate that aerobic heterotrophic bacteria within a uranium-contaminated environment that can hydrolyze organophosphate, especially in low pH conditions, may play an important role in the bioremediation of uranium

  5. Relation of fatty acid composition in lead-exposed mallards to fat mobilization, lipid peroxidation and alkaline phosphatase activity

    Mateo, R.; Beyer, W.N.; Spann, J.W.; Hoffman, D.J.

    2003-01-01

    The increase of n-6 polyunsaturated fatty acids (PUFA) in animal tissues has been proposed as a mechanism of Pb poisoning through lipid peroxidation or altered eicosanoids metabolism. We have studied fatty acid (FA) composition in liver and brain of mallards (Anas platyrhynchos) feeding for three weeks on diets containing combinations of low or high levels of vitamin E (20 or 200 UI/kg) and Pb (0 or 2 g/kg). Saturated FA, n-6 PUFA and total concentrations of FA were higher in livers of Pb-exposed mallards, but not in their brains. The percentage of n-6 PUFA in liver and brain was slightly higher in Pb-exposed mallards. The increase of n-6 PUFA in liver was associated with increased triglycerides and cholesterol in plasma, thus could be in part attributed to feed refusal and fat mobilization. The hepatic ratios between adrenic acid (22:4 n-6) and arachidonic acid (20:4 n-6) or between adrenic acid and linoleic acid (18:2 n-6) were higher in Pb exposed birds, supporting the existing hypothesis of increased fatty acid elongation by Pb. Among the possible consequences of increased n-6 PUFA concentration in tissues, we found increased lipid peroxidation in liver without important histopathological changes, and decreased plasma alkaline phosphatase activity that may reflect altered bone metabolism in birds.

  6. The maize (Zea mays ssp. mays var. B73 genome encodes 33 members of the purple acid phosphatase gene family

    Eliécer eGonzález Muñoz

    2015-05-01

    Full Text Available Purple acid phosphatases (PAPs play an important role in plant phosphorus nutrition, both by liberating phosphorus from organic sources in the soil and by modulating distribution within the plant throughout growth and development. Furthermore, members of the PAP protein family have been implicated in a broader role in plant mineral homeostasis, stress responses and development. We have identified 33 candidate PAP encoding gene models in the maize (Zea mays ssp. mays var. B73 reference genome. The maize Pap family includes a clear single-copy ortholog of the Arabidopsis gene AtPAP26, shown previously to encode both major intracellular and secreted acid phosphatase activities. Certain groups of PAPs present in Arabidopsis, however, are absent in maize, while the maize family contains a number of expansions, including a distinct radiation not present in Arabidopsis. Analysis of RNA-sequencing based transcriptome data revealed accumulation of maize Pap transcripts in multiple plant tissues at multiple stages of development, and increased accumulation of specific transcripts under low phosphorus availability. These data suggest the maize PAP family as a whole to have broad significance throughout the plant life cycle, while highlighting potential functional specialization of individual family members.

  7. Increased tartrate-resistant acid phosphatase (TRAP) expression in malignant breast, ovarian and melanoma tissue: an investigational study

    Tartrate-resistant acid phosphatase (TRAP) is a metalloprotein enzyme that belongs to the acid phosphatases and is known to be expressed by osteoclasts. It has already been investigated as a marker of bone metastases in cancer patients. In this study, which examined the value of serum TRAP concentrations as a marker of bone disease in breast cancer patients, we observed high concentrations of TRAP even in patients without bone metastases. To elucidate this phenomenon, we examined the expression of TRAP in breast cancer cells and the cells of several other malignancies. TRAP concentrations in the serum of tumor patients were determined by ELISA. The expression of TRAP in breast, ovarian, and cervical cancer and malignant melanoma was analyzed by immunohistochemistry. RT-PCR and immunocytology were used to evaluate TRAP expression in cultured tumor cells. A marked increase in serum TRAP concentrations was observed in patients with breast and ovarian cancer, regardless of the presence or absence of bone disease. TRAP expression was found in breast and ovarian cancers and malignant melanoma, while cervical cancer showed only minimal expression of TRAP. Expression of TRAP was absent in benign tissue or was much less marked than in the corresponding malignant tissue. TRAP expression was also demonstrated in cultured primary cancer cells and in commercially available cell lines. Overexpression of TRAP was detected in the cells of various different tumors. TRAP might be useful as a marker of progression of malignant disease. It could also be a potential target for future cancer therapies

  8. The effect of water and salt stresses on the phosphorus content and acid phosphatase activity in oilseed rape

    Stanisław Flasiński

    2014-02-01

    Full Text Available Oilseed rape plants responded to water and salt stresses (-0.5 MPa, PEG 6000 and NaCI by reduction of the fresh and dry weights of shoots and roots. When PEG was used, the ratio of dry weights of roots:shoots surpassed that of controls. The leaf protein content increased considerably. The phosphorus content decreased only in the roots, most significantly after three days of stress. Immediately after the stresses were induced, an increase in the acid phosphatase (AP activity was noted. Water and salt stresses caused four- and two-fold increases in AP activity in leaves, respectively. Changes in the enzyme activity were negligible in stems and roots. There are nine forms of AP in young leaves of oilseed rape. In the stressed plants, from No. 5 revealed lower activity and forms Nos 8 and 9, higher activities than in the control. The increase in AP activity was directly accompanied by the decrease in the water potential of the tissues. Oilseed rape is considerably less sensitive to salt stress than to water stress, which is manifested as the lower inhibition of plant growth and also by a smaller increase in acid phosphatase activity.

  9. Biosynthesis of acid phosphatase of baker's yeast . Characterization of a protoplast-bound fraction containing precursors of the exo-enzyme

    Boer, Pieter; Rijn, Herman J.M. van; Reinking, A.; Steyn-Parvé, Elizabeth P.

    1975-01-01

    1. 1.|Yest protoplasts, secreting acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum) EC 3.1.3.2) contain a small amount of firmly bound enzyme, even after lysis (Van Rijn, H.J.M.; Boer, P. and Steyn-Parvé, E.P. (1972) Biochim. Biophys. Acta 268, 431–441). The major part (70%

  10. Effect of x-irradiation on the acid and alkaline phosphatase activities in the south Indian scorpion Heterometrus fulvipes (C.L. Koch)

    Variations in the activity levels of acid phosphatase and alkaline phosphatase in the tissues of scorpion Heterometrus fulvipes after whole-body irradiation with X-rays are studied. The animals were exposed to sublethal dose (1/3 of LD50) of radiation for different periods of time ranging from 1/2h to 4h. Tissue homogenates were also irradiated and studied. The activity of phosphatases was found to decrease with increase in exposure time both at in vivo and in vitro, the per cent decrease being higher at in vitro. The activity levels of phosphatases in different tissues were in the order of hepato pancreas > heart pedipalpal > muscle > nervous tissue. (M.G.B.)

  11. Low Soil Phosphorus Availability Increases Acid Phosphatases Activities and Affects P Partitioning in Nodules, Seeds and Rhizosphere of Phaseolus vulgaris

    Jean-Jacques Drevon

    2012-06-01

    Full Text Available The effect of phosphorus (P deficiency on phosphatases activities in N2-fixing legumes has been widely studied in hydroponic culture. However, the response of acid phosphatase (APase and phytase in rhizosphere, nodules and seeds of Phaseolus vulgaris to low soil’s P-availability is not yet fully understood. In this study, six genotypes of N2-fixing P. vulgaris were grown under contrasting soil P-availabilities; i.e., low  (4.3 mg P kg−1 and sufficient (16.7 mg P kg−1 in the Haouz region of Morocco. At flowering and maturity stages, plants were harvested and analyzed for their phosphatases activities, growth and P content. Results show that, low P decreased nodulation, growth, P uptake and N accumulation in all the genotypes, but to a greater extent in the sensitive recombinant inbreed line 147. In addition, while seed P content was slightly reduced under low P soil; a higher P was noticed in the Flamingo and Contender large seeded-beans (6.15 to 7.11 mg g−1. In these latter genotypes, high APase and phytase activities in seeds and nodules were associated with a significant decline in rhizosphere’s available P. APase activity was mainly stimulated in nodules, whereas phytase activity was highly induced in seeds (77%. In conclusion, the variations of APase and phytase activities in nodules and seeds depend on genotype and can greatly influence the internal utilization of P, which might result in low P soil tolerance in N2-fixing legumes.

  12. Characterization of N-type glycosylation sites and glycan structures of Purple Acid Phosphatase Phytases from Wheat (Triticum aestivum L.)

    Dionisio, Giuseppe; Brinch-Pedersen, Henrik; Welinder, Karen Gjesing;

    2011-01-01

    Wheat (Triticum aestivum L.) possesses preformed phytase activity in the grain that is essential to make phosphate available to cell metabolism and in food and feed (Brejnholt S. et al., 2011). Cereals contain the purple acid phosphatase type of phytases, PAPhy (Dionisio G. et al., 2011a). Mature......) Cloning and Characterization of Purple Acid Phosphatase Phytases from Wheat (Triticum aestivum L.), Barley (Hordeum vulgare L.), Maize (Zea maize L.) and Rice (Oryza sativa L.). Plant Physiol. [in press, Jan 10, Epub ahead of print] Dionisio G., Brinch-Pedersen H., Welinder K.G., Jørgensen M. (2011b...

  13. Evaluating the levels of salivary alkaline and acid phosphatase activities as biochemical markers for periodontal disease: A case series

    Sarita Dabra

    2012-01-01

    Full Text Available Background: The purpose of this study was to determine the salivary levels of alkaline phosphatase (ALP and acid phosphatase (ACP activities in patients with periodontal disease and to evaluate the use of these enzymes as biochemical markers for periodontal tissue damage. Materials and Methods: In this prospective analytical study, we examined the activities of salivary ALP and ACP in patients with periodontal disease, before and after periodontal treatment. The experimental groups consisted of 20 gingivitis patients and 20 periodontitis patients and the control group had healthy subjects (20 samples. The stimulated saliva of the patient was collected in a sterile test tube and analyzed using Hitachi′s Diagnostic Automatic Analyser. Periodontal disease was determined based on clinical parameters such as gingival index, probing depth and clinical attachment loss. Patients with periodontal disease were under conventional periodontal treatment. The statistical analysis applied was Student′s t-test. Probabilities less than 0.05 (P < 0.05 were considered significant. Results: The obtained results showed statistically significant increased activities of ALP and ACP in saliva from patients with periodontal disease in relation to control group. A significant reduction in the enzyme levels was seen after conventional periodontal therapy. Conclusions: Based on these results, salivary ALP and ACP can be considered to be the biomarkers for evaluating periodontal tissue damage.

  14. Effect of inhibition of tyrosine phosphatases on voltage-operated calcium channel currents in rabbit isolated ear artery cells

    Wijetunge, S; Lymn, J S; Hughes, A.D.

    1998-01-01

    The effect of increasing cellular tyrosine phosphorylation by inhibiting endogenous tyrosine phosphatases was examined on voltage-operated calcium channel currents in vascular smooth muscle cells.In single ear artery smooth muscle cells of the rabbit, studied by the whole cell voltage clamp technique, intracellular application of the tyrosine phosphatase inhibitors, sodium orthovanadate (100 μM) and peroxyvanadate (100 μM orthovanadate+1 mM H2O2) increased voltage-operated calcium channel cur...

  15. Amylase and acid phosphatase activity in potato tubers treated with gibberellic acid and stored at 2°C and 8°C

    M. Bielińska-Czarnecka; K. Białek

    2015-01-01

    Amylase activity was higher in tubers stored at 2°C and mare marked in the soaked ones (both in water and in GA3). In the late and difficult-sprouting cv. Uran, sokaing resulted in increased amylolytic activity also at 8°C stored tubers. On the contrary, the acid phosphatase activity was a little higher at 8°C than at 2°C stored tubers. At the former temperature two peaks of activity were marked:, in November–December and February–March.

  16. Isolation and Characterization of Soil Fulvic Acid

    Mir Munsif Ali Talpur

    2016-06-01

    Full Text Available Fulvic acid was isolated from the agriculture soil of District Naushahro Feroz, Sindh, Pakistan by International Humic Substances Society (IHSS method. The nutrient contents of the soil like N. P, K, Ca, Mg, Fe and Zn were determined by using the Atomic Absorption spectrophotometer (AAS. The Spectroscopic analysis was carried out by studying the UV-Vis, FT-IR and NIR spectra of isolated compounds. The data has been compared with the literature and correlated. Moisture as well as texture shows good water holding capacity and silt- loam type of soil. pH and EC are indicators of the fertility of soil to be beneficial for plantation. The spectral data (UV-Visible, FTIR and NIR supports the characteristic functional groups (-COOH, C=O, -OH, -NH2, C=C, CH2 and Polysaccharides present in Fulvic acid. E4/E6 values depict its hydrophilic nature, having less aromatic and more aliphatic groups. The presence of metal ions indicates its chelating ability.

  17. Effect of noise exposure (85 dB ) on testicular adrenocortical steroidogenic key enzymes, acid and alkaline phosphatase activities of sex organs in mature albino rats

    2000-01-01

    Changes in the activities of △5-3β-hydroysteroid dehydrogenase (HSD) in testis and adrenal gland, 17β-hydroxysteroid dehydrogenase in testis, acid and alkaline phosphatase in testis, prostate and seminal vesicle were observed in noise exposed mature rats at the intensity of 85 dB for 8 h/day for 45 days. The results indicated that noise exposed group showed a significant diminution in the activities of androgenic key enzymes △5-3β and 17β-HSD, acid phosphatase in testis, prostate and seminal vesicle. There was a significant elevation in the activities of adrenal △5-3β-HSD, alkaline phosphatase in testis and other accessory sex organ in noise exposed group. Gonadosomatic, prostatosomatic and seminal vesiculo-somatic indexes were decreased significantly in noise exposed group. Therefore, it is evident that noise exposure at 85dB exerts a deleterious effect on testicular and adrenocortical activities.

  18. The effect of 1,25-dihydroxycholecalciferol on the multiple forms of alkaline phosphatase and the sialic acid incorporation into microsomes of chick duodenum

    Polyacrylamide disc gel electrophoresis of n-butanol solubilized alkaline phosphatase from chick duodenum revealed that the change of alkaline phosphatase induced by 1,25-(OH)2D3 involved the transformation of desialoenzyme to sialoenzyme. The initial stimulation by 1,25-(OH)2D3 of the incorporation of sialic acid into duodenal microsomes corresponded with the initial increase in calcium absorption. After this initial stimulation, there was a rapid decline in sialic acid incorporation into microsomes decreasing below control levels at 24 hr. Calcium concentration in the microsomes followed a pattern similar to the incorporation of sialic acid into microsomes. The depressed sialic acid incorporation was reversed by the addition of calcium in vitro. These results suggest that the initial action of 1,25-(OH)2D3 is to change the membrane permeability to calcium and to change the subcellular distribution of calcium in the small intestine. The accumulated calcium in the microsomes then stimulates the sialic acid incorporation into desialoenzyme. This results in the changes of isozyme pattern of alkaline phosphatase, viz, the transformation of desialoenzyme to sialoenzyme. The transformed alkaline phosphatase might be one of the factors involved more directly in the regulation of calcium transport in intestine. (auth.)

  19. Changes of the Biomass and Acid Phosphatase Activity in Maize (Zea mays L.) Lines Under Low-P Stress

    YAO Qilun

    2008-01-01

    A pot culture trial was conducted to investigate the changes of the biomass and acid phosphatase (APase) activity in 10 maize lines under low-P stress. P-deficiency significantly decreased the biomass, but induced the significant enhancement of the APase activity. Since P-deficiency had smaller effects on the low-P tolerant maize lines compared with P-sensitive lines, it was demonstrated that differences of tolerance to P-deficiency existed among 10 different maize lines. In addition, the relative biomass and APase activity changed during the vegetative stage of development, and there existed a significant correlation between the biomass and APase activity under low-P stress. These results suggest that the biomass and APase activity can be regarded as indicative traits of maize lines for tolerance to low-P stress at seedling stage.

  20. Increased tartrate-resistant acid phosphatase (TRAP expression in malignant breast, ovarian and melanoma tissue: an investigational study

    Eck M

    2006-07-01

    Full Text Available Abstract Background Tartrate-resistant acid phosphatase (TRAP is a metalloprotein enzyme that belongs to the acid phosphatases and is known to be expressed by osteoclasts. It has already been investigated as a marker of bone metastases in cancer patients. In this study, which examined the value of serum TRAP concentrations as a marker of bone disease in breast cancer patients, we observed high concentrations of TRAP even in patients without bone metastases. To elucidate this phenomenon, we examined the expression of TRAP in breast cancer cells and the cells of several other malignancies. Methods TRAP concentrations in the serum of tumor patients were determined by ELISA. The expression of TRAP in breast, ovarian, and cervical cancer and malignant melanoma was analyzed by immunohistochemistry. RT-PCR and immunocytology were used to evaluate TRAP expression in cultured tumor cells. Results A marked increase in serum TRAP concentrations was observed in patients with breast and ovarian cancer, regardless of the presence or absence of bone disease. TRAP expression was found in breast and ovarian cancers and malignant melanoma, while cervical cancer showed only minimal expression of TRAP. Expression of TRAP was absent in benign tissue or was much less marked than in the corresponding malignant tissue. TRAP expression was also demonstrated in cultured primary cancer cells and in commercially available cell lines. Conclusion Overexpression of TRAP was detected in the cells of various different tumors. TRAP might be useful as a marker of progression of malignant disease. It could also be a potential target for future cancer therapies.

  1. A study of acid phosphatase locus 1 in women with high fat content and normal body mass index.

    De Lorenzo, Antonino; Di Renzo, Laura; Puja, Alberto; Saccucci, Patrizia; Gloria-Bottini, Fulvia; Bottini, Egidio

    2009-03-01

    De Lorenzo and coworkers have recently described a class of women with normal body mass index (BMI) and high fat content (normal weight obese syndrome [NWO]). This observation prompted us to study the possible role of acid phosphatase locus 1 (ACP(1)) in the differentiation of this special class of obese subjects. Acid phosphatase locus 1 is a polymorphic gene associated with severe obesity and with total cholesterol and triglycerides levels. The enzyme is composed by 2 isoforms--F and S--that have different biochemical properties and probably different functions. The sample study was composed of 130 white women from the population of Rome. Total fat mass and percentage of fat mass were measured by dual-energy x-ray absorptiometry. Thirty-six women had a BMI less than 25 and percentage of fat mass greater than 30 (high fat, normal BMI [HFHB]), and 94 women showed a BMI greater than 25 and a percentage of fat mass greater than 30 (high fat, high BMI [HFHB]). In the whole sample, the proportion of low-activity ACP(1) genotypes (*A/*A and *B/*A) was higher than in controls. However, whereas HFNB showed a very high frequency of ACP(1) *A/*A genotype, high-fat, high-BMI women showed an increase of *B/*A genotype. These 2 genotypes differ in the concentration of F isoform and the F/S ratio, which are lower in ACP(1)*A/*A genotype than in ACP(1)*B/*A genotype. The genetic differentiation of the class of women with normal BMI and high fat content from the class showing a concordant level of the 2 parameters supports the hypothesis that HFNB class represents a special cluster of obese subjects not revealed by BMI evaluation. Because ACP(1) is present in adipocytes, the present observation suggests that F isoform may have a specific role in the regulation of quantity of adipose tissue. PMID:19217450

  2. Penostatin Derivatives, a Novel Kind of Protein Phosphatase 1B Inhibitors Isolated from Solid Cultures of the Entomogenous Fungus Isaria tenuipes

    Yu-Peng Chen

    2014-01-01

    Full Text Available Protein tyrosine phosphatase 1B (PTP1B is implicated as a negative regulator of insulin receptor (IR signaling and a potential drug target for the treatment of type II diabetes and other associated metabolic syndromes. Therefore, small molecular inhibitors of PTP1B can be considered as an attractive approach for the design of new therapeutic agents of type II diabetes diseases. In a continuing search for new protein phosphatase inhibitors from fungi, we have isolated a new compound, named penostatin J (1, together with three known ones, penostatin C (2, penostatin A (3, and penostatin B (4, from cultures of the entomogenous fungus Isaria tenuipes. The structure of penostatin J (1 was elucidated by extensive spectroscopic analysis. We also demonstrate for the first time that penostatin derivatives exhibit the best PTP1B inhibitory action. These findings suggest that penostatin derivatives are a potential novel kind of PTP1B inhibitors.

  3. CONTROL OF ALKALINE PHOSPHATASE ACTIVITY IN C3H10T1/2 CELLS: ROLE OF RETINOIC ACID AND CELL DENSITY

    The enzyme alkaline phosphatase (AP) has been shown to be lost or inappropriately expressed during carcinogenesis in some tissues. ecause retinoic acid (RA) appears to play a role in the normal regulation of the enzyme (RA up-regulates AP in a variety of cell types) we have sugge...

  4. Characterization of a soluble phosphatidic acid phosphatase in bitter melon (Momordica charantia)

    Momordica charantia is often called bitter melon, bitter gourd or bitter squash because its fruit has a bitter taste. The fruit has been widely used as vegetable and herbal medicine. Alpha-eleostearic acid is the major fatty acid in the seeds, but little is known about its biosynthesis. As an initia...

  5. Dietary free fatty acids form alkaline phosphatase-enriched microdomains in the intestinal brush border membrane

    Hansen, Gert H; Rasmussen, Karina; Niels-Christiansen, Lise-Lotte;

    2011-01-01

    Free fatty acids released during intralumenal digestion of dietary fat must pass through the enterocyte brush border membrane before triacylglycerol reassembly and subsequent chylomicron delivery to the lymph system. In the present work fluorescent BODIPY fatty acid analogs were used to study this......-linked enzyme is the membrane protein in the brush border with the highest affinity for lipid rafts, this implies that free fatty acids selectively insert stably into these membrane microdomains. We have previously shown that absorption of dietary lipids transiently induce a selective endocytosis of AP from the...... brush border, and from work by others it is known that fat absorption is accompanied by a rise in serum AP and secretion of surfactant-like particles from enterocytes. We propose that these physiological processes may be triggered by the sequestering of dietary free fatty acids in lipid raft...

  6. Valproic acid induces hair regeneration in murine model and activates alkaline phosphatase activity in human dermal papilla cells.

    Soung-Hoon Lee

    Full Text Available BACKGROUND: Alopecia is the common hair loss problem that can affect many people. However, current therapies for treatment of alopecia are limited by low efficacy and potentially undesirable side effects. We have identified a new function for valproic acid (VPA, a GSK3β inhibitor that activates the Wnt/β-catenin pathway, to promote hair re-growth in vitro and in vivo. METHODOLOGY/ PRINCIPAL FINDINGS: Topical application of VPA to male C3H mice critically stimulated hair re-growth and induced terminally differentiated epidermal markers such as filaggrin and loricrin, and the dermal papilla marker alkaline phosphatase (ALP. VPA induced ALP in human dermal papilla cells by up-regulating the Wnt/β-catenin pathway, whereas minoxidil (MNX, a drug commonly used to treat alopecia, did not significantly affect the Wnt/β-catenin pathway. VPA analogs and other GSK3β inhibitors that activate the Wnt/β-catenin pathway such as 4-phenyl butyric acid, LiCl, and BeCl(2 also exhibited hair growth-promoting activities in vivo. Importantly, VPA, but not MNX, successfully stimulate hair growth in the wounds of C3H mice. CONCLUSIONS/ SIGNIFICANCE: Our findings indicate that small molecules that activate the Wnt/β-catenin pathway, such as VPA, can potentially be developed as drugs to stimulate hair re-growth.

  7. The genetics of feto-placental development: A study of acid phosphatase locus 1 and adenosine deaminase polymorphisms in a consecutive series of newborn infants

    Bergamaschi Antonio

    2008-09-01

    Full Text Available Abstract Background Acid phosphatase locus 1 and adenosine deaminase locus 1 polymorphisms show cooperative effects on glucose metabolism and immunological functions. The recent observation of cooperation between the two systems on susceptibility to repeated spontaneous miscarriage prompted us to search for possible interactional effects between these genes and the correlation between birth weight and placental weight. Deviation from a balanced development of the feto-placental unit has been found to be associated with perinatal morbidity and mortality and with cardiovascular diseases in adulthood. Methods We examined 400 consecutive newborns from the Caucasian population of Rome. Birth weight, placental weight, and gestational length were registered. Acid phosphatase locus 1 and adenosine deaminase locus 1 phenotypes were determined by starch gel electrophoresis and correlation analysis was performed by SPSS programs. Informed verbal consent to participate in the study was obtained from the mothers. Results Highly significant differences in birth weight-placental weight correlations were observed among acid phosphatase locus 1 phenotypes (p = 0.005. The correlation between birth weight and placental weight was markedly elevated in subjects carrying acid phosphatase locus 1 phenotypes with medium-low F isoform concentration (A, CA and CB phenotypes compared to those carrying acid phosphatase locus 1 phenotypes with medium-high F isoform concentration (BA and B phenotypes (p = 0.002. Environmental and developmental variables were found to exert a significant effect on birth weight-placental weight correlation in subjects with medium-high F isoform concentrations, but only a marginal effect was observed in those with medium-low F isoform concentrations. The correlation between birth weight and placental weight is higher among carriers of the adenosine deaminase locus 1 allele*2, which is associated with low activity, than in homozygous adenosine

  8. Purification and characterization of a protein phosphatase (PP1-Arch) from the archaebacterium Sulfolobus solfataricus, isolation and expression of its gene

    Leng, Jie

    1994-01-01

    PP1-Arch was verified as a protein phosphatase by both acid molybdate extraction and thin layer electrophoresis. Soluble fraction was prepared from Sulfolobus solfataricus, from which PP1-Arch was purified over 1OOO-fold by DE-52 ion-exchange, hydroxyapatite, gel filtration (G- 100), and Mono Q FPLC chromatography. PP1-Arch was identified from the fmal purified sample by renaturation on an SDS-polyacrylamide gel. The molecular size of PP1-Arch was determined by both gel filtrat...

  9. A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

    Z. Wen

    2013-08-01

    Full Text Available Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer.

  10. Molecular Characterization and Functional Analysis of a New Acid Phosphatase Gene (Ha-acp1) from Heterodera avenae

    LIU Yan-ke; HUANG Wen-kun; LONG Hai-bo; PENG Huan; HE Wen-ting; PENG De-liang

    2014-01-01

    For sedentary endo-parasitic nematodes, parasitism genes encoding secretory protein expressed in the subventral glands cells always play an important role during the early parasitic process. A new acid phosphatase gene (Ha-acp1) expressed in the subventral glands of the cereal cyst nematode (Heterodera avenae) was cloned and the characteristics of the gene were analyzed. Results showed that the gene had a putative signal peptide for secretion and in situ hybridization showed that the transcripts of Ha-acp1 accumulated speciifcally in the subventral gland cells of H. avenae. Southern blot analysis suggested that Ha-acp1 belonged to a multigene family. RT-PCR analysis indicated that this transcription was strong at the pre-parasitic juveniles. Knocking down Ha-acp1 using RNA interference technology could reduce nematode infectivity by 50%, and suppress the development of cyst. Results indicated that Ha-acp1 could play an important role in destroying the defense system of host plants.

  11. A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

    Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D) models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer

  12. Root-exuded acid phosphatase and 32Pi-uptake kinetics of wheat, rye and triticale under phosphorus starvation

    A nutrient culture experiment was conducted with cereal species viz., wheat (Triticum aestivum L.) cv. PBW-343), rye (Secale cereale L cv. R-308) and triticale (Triticale octoploide L. cv. DT-46), a hybrid of wheat and rye, to examine the genetic variation in root-exuded acid phosphatase (ACPase) activity and kinetics of 32Pi-uptake under P deficient condition. The ACPase activity was assayed in the extract (intra-) and on surface (extra-cellular) or root, using p-nitrophenyl phosphate as substrate. Significantly higher ACPase activity was observed in wheat followed by rye and triticale both on the root surface and in root extract. In general, root surface ACPase activity was 2.2-fold higher than that in root extract. A strong correlation (r2 = 0.87**) between extra and intra-cellular ACPase activity was observed. In terms of kinetic parameters, it was observed that 32Pi uptake and Imax values were significantly higher in rye while Cmin and Km were lowest compared to wheat and triticale. The dry weights of shoot, root and total plant were significantly higher in rye compared to wheat and triticale. Rye also had higher amount of total plant P content The superiority of rye over wheat and triticale in P uptake was observed mainly due to efficient Pi-uptake system, which needs further studies to ascertain the enhancement of Pi-induced high-affinity P transporter in these cereals. (author)

  13. A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

    Wen, Z.; Liao, Q.; Hu, Y.; You, L.; Zhou, L.; Zhao, Y. [Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Tsinghua University, Beijing (China)

    2013-08-10

    Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D) models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer.

  14. Analysis of the contribution of acid phosphatase to P efficiency in Brassica napus under low phosphorus conditions

    2010-01-01

    To understand whether genotypic variation in acid phosphatase (APase) activity in rapeseed (Brassica napus L.) induced by phosphorus (P) deficiency has impact on P efficiency,soil APase activity in the rhizosphere for rapeseed P-efficient genotype 102 and P-inefficient genotype 105 was measured against organic and inorganic P sources in the pot experiment,and the activities of root-secreted APase and leaf intracellular APase were investigated in different P-starvation periods in the nutrient solution.Higher activity of root-secreted APase in B.napus was induced under low P conditions.However,P nutrition and P uptake efficiency of the plants supplied with organic P were not directly related to the activity of root-secreted APase due to several confounding factors affecting APase availability.The higher activity of leaf APase improved P remobilization in plants and played important roles in enhancing P use efficiency,shown by the significant correlation between leaf APase activity and P use efficiency in a rapeseed recombinant inbred population of 135 lines.

  15. Experimental studies on bone with focus on tartrate-resistant acid phosphatase and bone remodeling

    2014-01-01

    List of papers. Papers I and III are removed from the thesis due to publisher restrictions. Paper I : Melhus G, Solberg LB, Dimmen S, Madsen JE, Nordsletten L, Reinholt FP. Experimental osteoporosis induced by ovariectomy and vitamin D deficient diet does not markedly affect fracture healing in rats Acta Orthop 2007 Jun;78(3):393-403 doi:10.1080/17453670710013988 Paper II: Solberg LB, Brorson SH, Stordalen GA, Bækkevold E, Andersson G, Reinholt FP. Increased tartrate-resistant acid ph...

  16. The informative value of the radioimmunological determination of acid prostate phosphatase for the diagnostic of prostate carcinoma: A comparison of various reagent kits

    Patients of the university urological clinic in Freiburg gave, in all, 89 serum samples for the determination of acid prostate phosphatase using four different methods (3 radioimmunological methods and 1 enzymatic procedure) in order to compare the informative ability of these procedures in the diagnostic of prostate carcinoma. The results of the radioimmunological determinations agreed mostly with one another. They showed a higher sensitivity than the enzymatic method, but a lower specificity. The rate of false positive as well as false negative results is with both procedures too large, so that a reliable early diagnosis is not possible. Not until after metastasis of the carcinoma do all four methods give clearly pathological values. The acid prostate phosphatase determination is therefore suited for progress and therapy control. The values sink with successful therapy, rise again with recitivism. (TRV)

  17. In vivo detection of prostatic carcinoma with antibodies against prostatic acid phosphatase

    Serum prostatic acid phosphates (PAP) immunoassay is used to evaluate patients with prostatic carcinoma; however, as with other tumor markers, the enzyme levels do not necessarily reflect the presence or extent of tumor. The authors investigated the use of radiolabeled PAP antibodies for the in vivo detection of prostatic carcinoma by external scintillation imaging. Nine patients with prostatic carcinoma were entered into the study. Each received from 2.0 to 2.5 mCi of I-131 labeled antibody to PAP, administered i.v. The immunogen (PAP) was purified from normal human seminal fluid. Antiserum was prepared in rabbits by injecting the purified PAP. The antibodies were labeled with I-131 by chloramine-T method (10 to 20 Ci/g of IgG). Total body images were obtained at 24 and 48 hrs following administration of the labeled antibody. Nontarget I-131 activity was diminished by computer processing. Tumor sites detected by I-131 antibodies were correlated with other diagnostic procedures. In 7 of 9 patients primary and metastatic sites of cancer were detected by antibody imaging, however, no bone lesions were detected (6 cases). In 3 patients with concomitant pulmonary tumors, one was identified as of prostate origin. The serum PAP was normal in 4 patients; however, the primary tumor was identified in 3 of these. These findings suggest that the localization of prostatic carcinoma by means of in-vivo imaging of labeled antibodies to PAP is feasible and offers diagnostic opportunities based upon the functional characteristics

  18. Detection of Ca2+-dependent acid phosphatase activity identiifes neuronal integrity in damaged rat central nervous system after application of bacterial melanin

    Tigran R Petrosyan; Anna S Ter-Markosyan; Anna S Hovsepyan

    2016-01-01

    The study aims to confirm the neuroregenerative effects of bacterial melanin (BM) on central nervous system injury using a special staining method based on the detection of Ca2+-dependent acid phosphatase activity. Twenty-four rats were randomly assigned to undergo either unilateral destruction of sensorimotor cortex (group I;n=12) or unilateral rubrospinal tract transection at the cervical level (C3–4) (group II;n=12). In each group, six rats were randomly selected after surgery to undergo intramuscular injection of BM solution (BM subgroup) and the remaining six rats were intramuscularly injected with saline (saline subgroup). Neurological testing confirmed that BM accelerated the recovery of motor function in rats from both BM and saline subgroups. Two months after surgery, Ca2+-dependent acid phosphatase activity detection in combination with Chilingarian’s calcium adenoside triphosphate method revealed that BM stimulated the sprouting of ifbers and dilated the capillaries in the brain and spinal cord. These results sug-gest that BM can promote the recovery of motor function of rats with central nervous system injury;and detection of Ca2+-dependent acid phosphatase activity is a fast and easy method used to study the regenera-tion-promoting effects of BM on the injured central nervous system.

  19. Detection of Ca2+-dependent acid phosphatase activity identifies neuronal integrity in damaged rat central nervous system after application of bacterial melanin

    Tigran R Petrosyan

    2016-01-01

    Full Text Available The study aims to confirm the neuroregenerative effects of bacterial melanin (BM on central nervous system injury using a special staining method based on the detection of Ca2+-dependent acid phosphatase activity. Twenty-four rats were randomly assigned to undergo either unilateral destruction of sensorimotor cortex (group I; n = 12 or unilateral rubrospinal tract transection at the cervical level (C3–4 (group II; n = 12. In each group, six rats were randomly selected after surgery to undergo intramuscular injection of BM solution (BM subgroup and the remaining six rats were intramuscularly injected with saline (saline subgroup. Neurological testing confirmed that BM accelerated the recovery of motor function in rats from both BM and saline subgroups. Two months after surgery, Ca2+-dependent acid phosphatase activity detection in combination with Chilingarian's calcium adenoside triphosphate method revealed that BM stimulated the sprouting of fibers and dilated the capillaries in the brain and spinal cord. These results suggest that BM can promote the recovery of motor function of rats with central nervous system injury; and detection of Ca2+-dependent acid phosphatase activity is a fast and easy method used to study the regeneration-promoting effects of BM on the injured central nervous system.

  20. Fatty acid synthase inhibitors isolated from Punica granatum L

    Jiang, He-Zhong [School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, (China); Ma, Qing-Yun; Liang, Wen-Juan; Huang, Sheng-Zhuo; Dai, Hao-Fu; Wang, Peng-Cheng; Zhao, You-Xing, E-mail: zhaoyx1011@163.com [Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou (China); Fan, Hui-Jin; Ma, Xiao-Feng, E-mail: maxiaofeng@gucas.ac.cn [College of Life Sciences, Graduate University of Chinese Academy of Sciences, Beijing (China)

    2012-05-15

    The aim of this work is the isolation of fatty acid synthase (FAS) inhibitors from the ethyl acetate extracts of fruit peels of Punica granatum L. Bioassay-guided chemical investigation of the fruit peels resulted in the isolation of seventeen compounds mainly including triterpenoids and phenolic compounds, from which one new oleanane-type triterpene (punicaone) along with fourteen known compounds were isolated for the first time from this plant. Seven isolates were evaluated for inhibitory activities of FAS and two compounds showed to be active. Particularly, flavogallonic acid exhibited strong FAS inhibitory activity with IC{sub 50} value of 10.3 {mu}mol L{sup -1}. (author)

  1. Fatty acid synthase inhibitors isolated from Punica granatum L

    The aim of this work is the isolation of fatty acid synthase (FAS) inhibitors from the ethyl acetate extracts of fruit peels of Punica granatum L. Bioassay-guided chemical investigation of the fruit peels resulted in the isolation of seventeen compounds mainly including triterpenoids and phenolic compounds, from which one new oleanane-type triterpene (punicaone) along with fourteen known compounds were isolated for the first time from this plant. Seven isolates were evaluated for inhibitory activities of FAS and two compounds showed to be active. Particularly, flavogallonic acid exhibited strong FAS inhibitory activity with IC50 value of 10.3 μmol L-1. (author)

  2. EFEITO DE FATORES AMBIENTAIS DA FOSFATASE ÁCIDA NO FEIJOEIRO EFFECTS OF ENVIRONMENTAL FACTORS ON THE ACTIVITY OF ACID PHOSPHATASE IN COMMON BEAN

    José Renato de Freitas

    2007-09-01

    Full Text Available

    Plantas com 15 dias após a germinação foram colhidas em experimentos de campo com a finalidade de conhecer o pH, temperatura e tempo necessários para melhor expressar a atividade da fosfatase ácida em três variedades do feijoeiro (Phaseolus vulgaris L., Carioca, EMP-84 e CNF-l0, na presença e na ausência de fósforo. Os maiores valores de atividade da fosfatase ácida foram observadas quando as plantas foram colocadas em solução em pH 5,5 durante 120 minutos à temperatura de 30°C. A utilização de substâncias tamponantes como PNPP + Triton X-100 expressaram melhor a atividade da fosfatase ácida. As condições de vácuo constituíram um fator positivo para a atividade da fosfatase ácida. As plantas desenvolvidas sob estresse hídrico apresentaram menor atividade da fosfatase ácida. A relação folha-raiz da atividade da fosfatase ácida atingiu 5,72 para a variedade Carioca, 4,91 para a variedade EMP-84 e 4,36 para a variedade CNF-10.

    PALAVRAS-CHAVE: pH; temperatura; solução tamponada; tempo de reação; Phaseolus vulgaris.

    Plants with 15 days after the germination were picked in field experiments with the purpose of knowing the best pH, temperature and the necessary time to express the activity of the phosphatase acid in three bean varieties (Phaseolus vulgaris L., Carioca, EMP-84 and CNF-10, in the presence and in the phosphorus absence. The largest values of activity of the phosphatase acid were observed when the plants were tested in pH 5.5 solution during 120 minutes at the temperature of 30°C. The use of buffer substances as PNPP + Triton X-100 expressed better the activity of the phosphatase acid. The vacuum condition constituted a positive factor to express the activity of the phosphatase acid. The plants

  3. Tannic acid degradation by Klebsiella strains isolated from goat feces

    Arezoo Tahmourespour

    2016-03-01

    Full Text Available Background and Objectives: Tannins are toxic polyphenols that either bind and precipitate or condense proteins. The high tannin content of some plants is the preliminary limitation of using them as a ruminant feed. So, the aim of this study was the isolation and characterization of tannic acid degrading bacterial strains from goat feces before and after feeding on Pis- tachio-Soft Hulls as tannin rich diet (TRD.Materials and Methods: Bacterial strains capable of utilizing tannic acid as sole carbon and energy source were isolated and characterized from goat feces before and after feeding on TRD. Tannase activity, maximum tolerable concentration and biodegradation potential were assessed.Results: Four tannase positive isolates were identified as Klebsiella pneumoniae. Isolated strains showed the maximum tolerable concentration of 64g/L of tannin. The tannic acid degradation percentage at a concentration of 15.0 g/L reached a maximum of 68% after 24 h incubation, and more than 98% after 72 h incubation. The pH of the medium also decreased along with tannic acid utilization.Conclusions: It is obvious that TRD induced adaptive responses. Thus, while the bacteria were able to degrade and detoxify the tannic acids, they had to adapt in the presence of high concentrations of tannic acid. So, these isolates have an amazing potential for application in bioremediation, waste water treatment, also reduction of tannins antinutritional effects in animal feeds.Keywords: Biodegradation; Goat feces; Klebsiella strains; Tannic acid

  4. Isolation of deoxyribonucleic acids (A Review)

    The criteria of choice in this Review have been to gather some of the last advances in the methodology of DNAs isolation; also the description of the generally accepted procedures has been emphasized. Only papers published before March 1974 are reviewed, because this work has been finished during this month. (Author) 109 refs

  5. Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential.

    Mohd Adnan, Ahmad Faris; Tan, Irene K P

    2007-05-01

    Two traditional fermented food 'tapai' (fermented tapioca) and 'tempoyak' (fermented durian flesh), chilli puree and fresh goat's milk were used as sources for the isolation of lactic acid bacteria (LAB). A total of 126 isolates were obtained and by sequential screening for catalase activity and Gram-staining, 55 were determined to be LAB out of which 16 were established to be homofermentative by the gel plug test. Seven isolates were identified by use of the API 50CHL kit and two lactobacilli strains and one lactococci strain were selected to study their growth and lactic acid production profiles in a time course experiment. The lactobacilli strains, both isolated from 'tapai', produced higher amounts of cells and lactic acid from glucose as compared to the lactococci strain isolated from fresh goat's milk. PMID:16872826

  6. The myeloperoxidase-derived oxidant hypothiocyanous acid inhibits protein tyrosine phosphatases via oxidation of key cysteine residues

    Cook, Naomi L.; Moeke, Cassidy H.; Fantoni, Luca I.;

    2016-01-01

    Phosphorylation of protein tyrosine residues is critical to cellular processes, and is regulated by kinases and phosphatases (PTPs). PTPs contain a redox-sensitive active site Cys residue, which is readily oxidized. Myeloperoxidase, released from activated leukocytes, catalyzes thiocyanate ion (S...

  7. Rapid typing of herpes simplex virus isolates by deoxyribonucleic acid:deoxyribonucleic acid hybridization.

    Brautigam, A R; Richman, D D; Oxman, M N

    1980-01-01

    A method for typing clinical isolates of herpes simplex virus was developed. It utilizes hybridization between unlabeled deoxyribonucleic acid from infected cultures and tritium-labeled virus deoxyribonucleic acid, and it can be completed within a day using a single roller-tube culture of the clinical isolated. The data obtained are inherently quantitative, and the method yields unequivocal identification and typing. Thirty-nine coded clinical isolates were all correctly typed by this method.

  8. Molecular cloning of rat type 2C (IA) protein phosphatase mRNA.

    Tamura, S; Lynch, K. R.; Larner, J.; Fox, J.; Yasui, A; Kikuchi, K.; Suzuki, Y.; Tsuiki, S

    1989-01-01

    A full-length cDNA encoding rat type 2C (IA) protein phosphatase was isolated from a kidney cDNA library. The cDNA was identified by screening the library with oligonucleotides based on a partial amino acid sequence determined from purified rat liver phosphatase. This clone is 2.35 kilobase pairs long and has a single extended translation reading frame that predicts a 382-amino acid protein of 42,416 daltons. The deduced amino acid sequence contains segments corresponding to three peptides fr...

  9. Fosfatasa ácida en Oxisoles bajo cultivo de tabaco Acid phosphatase in Oxisols under tobacco cropping

    Toledo Marcela

    2010-07-01

    Full Text Available En suelos ácidos de trópicos y subtrópicos, caracterizados por una baja disponibilidad de P para las plantas, el papel de las fosfatasas ácidas en la mineralización del P orgánico es fundamental, constituyendo una variable promisoria para estimar la calidad del suelo. El objetivo del trabajo fue evaluar la actividad de la fosfatasa ácida en Oxisoles bajo uso tabacalero, como indicador sensible de calidad. En la provincia de Misiones ubicada al nordeste de la República Argentina, se estableció un ensayo sobre Eutrudoxes Ródicos, familia arcillosa fina, hipertérmica, aplicándose un diseño con cuatro bloques completos aleatorizados. Se establecieron 2 tratamientos: selva subtropical (Sv y uso tabacalero (Ta. Se tomaron muestras compuestas a 3 profundidades: 0-10; 10-20; 20-30 cm. Se determinaron las siguientes variables: actividad de la fosfatasa ácida (APA, pH, contenido de arcilla, carbono orgánico edáfico (CO, nitrógeno total (N, fósforo asimilable (P, materia orgánica particulada (MOP, y respiración del suelo (RES. En los casos estudiados, la APA fue mayor en los primeros diez centímetros de suelo, y fue disminuyendo con el aumento de la profundidad del perfil, en estrecha relación con los contenidos orgánicos del suelo. El 70% de la variabilidad de la APA se explicó por el nitrógeno total, íntimamente relacionado con la materia orgánica del suelo (pSoil biological parameters are of great value as sensitive indicators of transformations occurring under different uses and management practices (Mijangos et al., 2006. The aim of this study was to evaluate the activity of the acid phosphatase enzyme in Oxisols under tobacco cropping. The experimental design was in randomized complete blocks, with two treatments: subtropical rainforest (Sv and tobacco cropping (Ta (Nicotiana tabacum L.. Soil samples were taken from 0-10, 10 -20 and 20 -30 cm-deep layers. The variables measured were: APA, pH, clay content, total nitrogen (N

  10. CHARACTERIZATION OF LACTIC ACID BACTERIA ISOLATED FROM SUMBAWA MARE MILK

    Nengah Sujaya

    2008-06-01

    Full Text Available A study was carried out to isolate and characterize lactic acid bacteria (LAB from the Sumbawa mares milk The Isolation of LAB was conducted in Man Rogosa Sharpe (MRS agar. The isolates were characterized by standard methods, such as Gram staining, cell morphology study and fermentation activities. The ability of the isolates to inhibit some pathogenic bacteria was studied by dual culture assay. Isolates showing the widest spectrum of inhibiting pathogenic bacteria were further identified using API 50 CHL. The results showed that Sumbawa mare milk was dominated by lactobacilli and weisella/leuconostoc. As many as 26 out 36 isolates belong to homofermentative lactobacilli and another 10 isolates belong to both heterofermentative lactobacilli and weissella or leuconostoc. Twenty four isolates inhibited the growth of Escherichia coli 25922, Shigela flexneri, Salmonella typhimurium, and Staphylococcus aureus 29213. Two promising isolates with the widest spectrum of inhibiting pathogenic bacteria, Lactobacillus sp. SKG34 and Lactobacillus sp. SKG49, were identified respectively as Lactobacillus rhamnosus SKG34 and Lactobacillus ramnosus SKG49. These two isolates were specific strains of the sumbawa mare milk and are very potential to be developed as probiotic for human.

  11. Is the release of acid phosphatases by ectomycorrhizal fungi a matter of environmental conditions or species in situ?

    Plassard, Claude; Ali, Muhhammad A.; Duchemin, Myriam; Legname-Vonarx, Elvira; Louche, Julien; Cloutier-Hurteau, Benoît

    2011-01-01

    Ectomycorrhizal (ECM) fungi are able to release significant amounts of phosphatases (Pases) in their environment. These enzymes, by releasing the phosphate group of organic P, may play an important role in the recycling of P in forest soil. However, whether this enzyme release depends on the fungal diversity and / or the nutrient availability is not known. We addressed this question in the maritime pine (Pinus pinaster) forest, the first planted area in France on sandy podzol very poor in ...

  12. Screening on Gibberellic Acid Producing Activity of Azospirillum Isolates

    Six strains of Azopirillum spp were isolated from rice, sugarcane, corn, maize, sunflower and pepper roots and screened the gibberellic acid productivity. Only three strains of Azospirillum species showed the activity and were indentified by cultural, biochemical and drug sensitivity patterns. Among them,one strain isolated from rice root can produce microbial gibberellic acid. It showed greenish yellow colour in chromatogram under UV absorption. This screening method was studied from 1 to 14 days incubation. Qualitative measurement of GA productivity was determined by thin layer chromatography.

  13. Bioconversion of ferulic acid to vanillic acid by Halomonas elongata isolated from table-olive fermentation

    Abdelkafi, Slim; Sayadi, S.; Ben Ali Gam, Zouhaier; Casalot, Laurence; Labat, Marc

    2006-01-01

    Halomonas elongata strain Mar (=CCUG 52759) isolated from table-olive fermentation is the first halophilic bacterium to be shown to transform ferulic acid to vanillic acid under hypersaline conditions. During growth on ferulic acid, this strain was capable of promoting the formation of a significant amount of vanillic acid and trace quantities of vanillin. The products were confirmed by high-performance liquid chromatography and gas chromatography-mass spectrometry analyses. Based on the diff...

  14. Effect of andrographolide on phosphatases activity and cytotoxicity against Spodoptera litura

    Edwin, E.; P Vasantha-Srinivasan; A Thanigaivel; A Ponsankar; S Selin-Rani; K. Kalaivani; WB Hunter; Duraipandiyan, V; NA AlDhabi

    2016-01-01

    Andrographolide was isolated from ethanol extraction of Andrographis paniculata by column chromatography. Evaluation of larvicidal efficacy, enzymatic changes and cytotoxic activities against Spodoptera litura were conducted across a range of concentrations. The compound showed significant larvicidal activity between 5 - 25 ppm, post ingestion. Morphological deformities observed in larval-pupal stages. Enzymatic profiles were altered by reduction in acid phosphatase, ACP activity ...

  15. The Role of DmCatD, a Cathepsin D-Like Peptidase, and Acid Phosphatase in the Process of Follicular Atresia in Dipetalogaster maxima (Hemiptera: Reduviidae), a Vector of Chagas' Disease

    Leyria, Jimena; Fruttero, Leonardo L.; Nazar, Magalí; Canavoso, Lilián E.

    2015-01-01

    In this work, we have investigated the involvement of DmCatD, a cathepsin D-like peptidase, and acid phosphatase in the process of follicular atresia of Dipetalogaster maxima, a hematophagous insect vector of Chagas’ disease. For the studies, fat bodies, ovaries and hemolymph were sampled from anautogenous females at representative days of the reproductive cycle: pre-vitellogenesis, vitellogenesis as well as early and late atresia. Real time PCR (qPCR) and western blot assays showed that DmCatD was expressed in fat bodies and ovaries at all reproductive stages, being the expression of its active form significantly higher at the atretic stages. In hemolymph samples, only the immunoreactive band compatible with pro-DmCatD was observed by western blot. Acid phosphatase activity in ovarian tissues significantly increased during follicular atresia in comparison to pre-vitellogenesis and vitellogenesis. A further enzyme characterization with inhibitors showed that the high levels of acid phosphatase activity in atretic ovaries corresponded mainly to a tyrosine phosphatase. Immunofluorescence assays demonstrated that DmCatD and tyrosine phosphatase were associated with yolk bodies in vitellogenic follicles, while in atretic stages they displayed a different cellular distribution. DmCatD and tyrosine phosphatase partially co-localized with vitellin. Moreover, their interaction was supported by FRET analysis. In vitro assays using homogenates of atretic ovaries as the enzyme source and enzyme inhibitors demonstrated that DmCatD, together with a tyrosine phosphatase, were necessary to promote the degradation of vitellin. Taken together, the results strongly suggested that both acid hydrolases play a central role in early vitellin proteolysis during the process of follicular atresia. PMID:26091289

  16. Polyunsaturated fatty acids production by Schizochytrium sp. isolated from mangrove

    K.W. Fan

    2003-09-01

    Full Text Available Five Schizochytrium strains (N-1, N-2, N-5, N-6, and N-9 were isolated from fallen, senescent leaves of mangrove tree (Kandelia candel in Hong Kong. The fungi were cultivated in glucose yeast extract medium containing 60 g of glucose, 10 g of yeast extract and 1 L of 15‰ artificial seawater, initial pH 6.0, with shaking for 52 hr at 25ºC. Biomass yields of 5 isolates ranged from 10.8 to 13.2 g/l. Isolate N-2 yielding the highest dried cell mass at 13.2 g/l and isolate N-9 grew poorly with 10.8 g/l of biomass. EPA (Eicosapentaenoic acid, 20:5n-3 yield was low in most strains, while DHA (Docosahexaenoic acid, 22:6n-3 was high on the same medium. The contents of DHA in biomass varied: 174.9, 203.6, 186.1, 171.3 and 157.9 mg/g of dried-biomass for Schizochytrium isolate N-1, N-2, N-5, N-6, and N-9, respectively. Isolate N-2 had the highest proportion of DHA in fatty acid profile with 15:0, 28.7%; 16:0, 21.3%; 18:0, 0.9%; 18:3, 0.2%; 20:4, 0.3%; 20:5, 0.9%; 22:4, 6.7%; 22:6, 36.1%; and others, 9.3%. The salinity range for growth of Schizochytrium isolates was from 0-30‰ with optimum salinity for growth between 20-30‰.

  17. Reduction of Isolation Period of Coal Humic Acids

    Claudia H.H. Chen; Osumanu H. Ahmed; Nik M.A. Majid; Mohamadu B. Jalloh

    2009-01-01

    Problem statement: The isolation of Humic Acids (HA) from coal is laborious, costly and time consuming. The extraction and fractionation periods of HA vary from 4 h to 7 days. Fractionation period ranges from 12-24 h. However, most studies use 24 h as extraction period and also 24 h as fractionation period. This study was conducted to investigate whether the isolation period for HA of coals could be reduced. Approach: Different extraction periods using 0.1 M NaOH (4, 8, 12, 16, 20 and 24 h) w...

  18. Complex coordinated extracellular metabolism: Acid phosphatases activate diluted human leukocyte proteins to generate energy flow as NADPH from purine nucleotide ribose.

    Hibbs, John B; Vavrin, Zdenek; Cox, James E

    2016-08-01

    Complex metabolism is thought to occur exclusively in the crowded intracellular environment. Here we report that diluted enzymes from lysed human leukocytes produce extracellular energy. Our findings involve two pathways: the purine nucleotide catabolic pathway and the pentose phosphate pathway, which function together to generate energy as NADPH. Glucose6P fuel for NADPH production is generated from structural ribose of purine ribonucleoside monophosphates, ADP, and ADP-ribose. NADPH drives glutathione reductase to reduce an oxidized glutathione disulfide-glutathione redox couple. Acid phosphatases initiate ribose5P salvage from purine ribonucleoside monophosphates, and transaldolase controls the direction of carbon chain flow through the nonoxidative branch of the pentose phosphate pathway. These metabolic control points are regulated by pH. Biologically, this energy conserving metabolism could function in perturbed extracellular spaces. PMID:26895212

  19. Naturally Occurring Lactic Acid Bacteria Isolated from Tomato Pomace Silage

    Wu, Jing-Jing; Du, Rui-ping; Gao, Min; Sui, Yao-qiang; Xiu, Lei; Wang, Xiao

    2014-01-01

    Silage making has become a significant method of forage conservation worldwide. To determine how tomato pomace (TP) may be used effectively as animal feed, it was ensilaged for 90 days and microbiology counts, fermentation characteristics and chemical composition of tomato pomace silage (TPS) were evaluated at the 30th, 60th, and 90th days, respectively. In addition, 103 lactic acid bacteria were isolated from TPS. Based on the phenotypic and chemotaxonomic characteristics, 16S rDNA sequence ...

  20. Intravenous amino acids in third trimester isolated oligohydramnios

    To determine the efficacy of maternal administration of intravenous amino acid solution in improving amniotic fluid volume in cases of isolated oligohydramnios and to observe its impact on mode of delivery and neonatal outcome. Study Design: A prospective case series. Methodology: Forty two women with singleton pregnancy, well established gestational age and clinically and sonographically proven isolated oligohydramnios in the third trimester before 36 weeks were administered amino acid solution intravenously after excluding cases of premature rupture of membranes, congenital anomaly of fetus, maternal pulmonary, cardiovascular and hypertensive disorders, and severe placental insufficiency (raised S/D ratio). Pre-infusion and postinfusion Amniotic fluid Index (AFI) was measured and repeated weekly. Women were followed till delivery. Results: According to repeated measurement analysis of variance, mean pre-infusion AFI was 4.7 cm, mean one week postinfusion AFI was 5.8 cm, mean two week post-infusion AFI was 6.2 cm and mean three week AFI was 6.3 cm (p-value 0.029, significant). Cesarean section became a predominant mode of delivery in this group without a firm evidence of associated fetal compromise. Conclusion: Amino acid infusion is an effective therapy for raising AFI in isolated oligohydramnios in this case series. Liberal use of cesarean section in this selected group should be carefully re-evaluated. (author)

  1. Isolation of phosphatase-producing phosphate solubilizing bacteria from Loriya hot spring: Investigation of phosphate solubilizing in the presence of different parameters

    Maryam Parhamfar

    2014-04-01

    Full Text Available Introduction: Biofertilizers are the microorganisms that can convert useless nutrient to usable compounds. Unlike fertilizer, cost of biofertilizer production is low and doesn’t produce ecosystem pollution. Phosphate fertilizers can be replaced by phosphate biofertilizer to produce improvement. So, it is necessary to screen the climate-compatible phosphate solubilizing bacteria. Materials and methods: In this project samples were picked up from Loriya hot spring, which are located in Jiroft. Samples were incubated in PKV medium for 3 days. Screening of phosphate solubilizing bacteria was performed on the specific media, based on clear area diameter. The best bacterium was identified based on 16s rDNA gene. Phosphate solubilizing activity of this strain was considered in different carbon, nitrogen, phosphate and pH sources. Results: Sequence alignment and phylogenetic tree results show that B. sp. LOR033 is closely related to Bacillus licheniformis, with 97% homology. In addition, results show that maximum enzyme production was performed after 2 days that incubation pH was decreased simultaneously when the time was increased. Carbon sources investigation show that glucose is the most appropriate in enzyme production and phosphate releasing. Furthermore, results show that the optimum initial pH for phytase production was pH5.0. Different phosphate sources show that tricalcium phosphate has the suitable effect on enzyme activity in three days of incubation. Discussion and conclusion: Phosphatase enzyme production capacity, growth in acidic pH and phosphate solubilizing potential in different salt and phosphate sources show that this strain has considerable importance as biofertilizers.

  2. Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acid-etched (MSLA), laser-treated, and laser and acid-treated Ti surfaces

    Li, Lin-Jie; Kim, So-Nam

    2016-01-01

    PURPOSE In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P.05). CONCLUSION This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies. PMID:27350860

  3. The Tinkerbell (Tink) Mutation Identifies the Dual-Specificity MAPK Phosphatase INDOLE-3-BUTYRIC ACID-RESPONSE5 (IBR5) as a Novel Regulator of Organ Size in Arabidopsis

    Johnson, Kim L.; Ramm, Sascha; Kappel, Christian; Ward, Sally; Leyser, Ottoline; Sakamoto, Tomoaki; Kurata, Tetsuya; Bevan, Michael W.; Lenhard, Michael

    2015-01-01

    Mitogen-activated dual-specificity MAPK phosphatases are important negative regulators in the MAPK signalling pathways responsible for many essential processes in plants. In a screen for mutants with reduced organ size we have identified a mutation in the active site of the dual-specificity MAPK phosphatase INDOLE-3-BUTYRIC ACID-RESPONSE5 (IBR5) that we named tinkerbell (tink) due to its small size. Analysis of the tink mutant indicates that IBR5 acts as a novel regulator of organ size that c...

  4. Spatial structure of oligopeptide PAP(248-261), the N-terminal fragment of the HIV enhancer prostatic acid phosphatase peptide PAP(248-286), in aqueous and SDS micelle solutions

    Blokhin, Dmitriy S.; Filippov, Andrei V.; Antzutkin, Oleg N.; Karataeva, Farida Kh.; Klochkov, Vladimir V.

    2014-07-01

    Prostatic acid phosphatase (PAP) is an enzyme that facilitates infection of cells by HIV. Its peptide fragment PAP(248-286) forms amyloid fibrils known as SEVI, which enhance attachment of the virus by viral adhesion to the host cell prior to receptor-specific binding via reducing the electrostatic repulsion between the membranes of the virus and the target cell. The secondary structure of PAP(248-286) in aqueous and SDS solutions can be divided into an N-terminal disordered region, an α-helical central part and an α/310-helical C-terminal region (Nanga et al., 2009). In this work, we used NMR spectroscopy to study the spatial structure of the isolated N-terminal fragment of PAP(248-286), PAP(248-261) (GIHKQKEKSRLQGG), in aqueous and SDS micelle solutions. Formation of a PAP(248-261)-SDS complex was confirmed by chemical shift alterations in the 1H NMR spectra of the peptide, as well as by the signs and values of Nuclear Overhauser Effect (NOE). In addition, the PAP(248-261) peptide does not form any specified secondary structure in either aqueous or SDS solutions.

  5. Reduction of Isolation Period of Coal Humic Acids

    Claudia H.H. Chen

    2009-01-01

    Full Text Available Problem statement: The isolation of Humic Acids (HA from coal is laborious, costly and time consuming. The extraction and fractionation periods of HA vary from 4 h to 7 days. Fractionation period ranges from 12-24 h. However, most studies use 24 h as extraction period and also 24 h as fractionation period. This study was conducted to investigate whether the isolation period for HA of coals could be reduced. Approach: Different extraction periods using 0.1 M NaOH (4, 8, 12, 16, 20 and 24 h were tested. Samples were centrifuged (16,211 G for 15 min at the end of each extraction period. The dark-colored supernatant liquor containing HA was decanted and the pH of solution was adjusted to 1.0 using 6 M HCl. After acidification, the fractionation periods evaluated were 4, 8, 12, 16, 20 and 24 h. The samples were transferred to a polyethylene bottle and centrifuged (16, 211 G for 10 min after each fractionation period. The HA purification was done by suspending them in 50 mL distilled water and centrifuged (16,211 G for 10 min. HA samples were dried in an oven at 40°C to a constant weight. Standard procedures were used to characterized the HA (total carbon, E4/E6, phenolic OH, carboxylic COOH and total acidity. Results: There was significant effect of both extraction and fractionation periods on the isolation of HA from coal. The optimum period for Na ions to saturate the exchange complex of HA during the extraction process was 8 h while the optimum period for the exchanges sites of the HA to be saturated with H ions during the fractionation process was 20 h. The distilled water used in this study was able to purify HA within 1 h because it served as Bronsted-Lowry acid. Additionally, carbon, E4/E6, phenolic OH, carboxylic COOH and total acidity of the HA were typical of those reported in the literature, suggesting that that the isolation process of the HA was successful. Conclusion: The isolation period of HA from coal can be reduced to 29 h (8 h

  6. Naturally occurring lactic Acid bacteria isolated from tomato pomace silage.

    Wu, Jing-Jing; Du, Rui-Ping; Gao, Min; Sui, Yao-Qiang; Xiu, Lei; Wang, Xiao

    2014-05-01

    Silage making has become a significant method of forage conservation worldwide. To determine how tomato pomace (TP) may be used effectively as animal feed, it was ensilaged for 90 days and microbiology counts, fermentation characteristics and chemical composition of tomato pomace silage (TPS) were evaluated at the 30th, 60th, and 90th days, respectively. In addition, 103 lactic acid bacteria were isolated from TPS. Based on the phenotypic and chemotaxonomic characteristics, 16S rDNA sequence and carbohydrate fermentation tests, the isolates were identified as 17 species namely: Lactobacillus coryniformis subsp. torquens (0.97%), Lactobacillus pontis (0.97%), Lactobacillus hilgardii (0.97%), Lactobacillus pantheris (0.97%), Lactobacillus amylovorus (1.9%), Lactobacillus panis (1.9%), Lactobacillus vaginalis (1.9%), Lactobacillus rapi (1.9%), Lactobacillus buchneri (2.9%), Lactobacillus parafarraginis (2.9%), Lactobacillus helveticus (3.9%), Lactobacillus camelliae (3.9%), Lactobacillus fermentum (5.8%), Lactobacillus manihotivorans (6.8%), Lactobacillus plantarum (10.7%), Lactobacillus harbinensis (16.5%) and Lactobacillus paracasei subsp. paracasei (35.0%). This study has shown that TP can be well preserved for 90 days by ensilaging and that TPS is not only rich in essential nutrients, but that physiological and biochemical properties of the isolates could provide a platform for future design of lactic acid bacteria (LAB) inoculants aimed at improving the fermentation quality of silage. PMID:25049999

  7. Naturally Occurring Lactic Acid Bacteria Isolated from Tomato Pomace Silage

    Wu, Jing-jing; Du, Rui-ping; Gao, Min; Sui, Yao-qiang; Xiu, Lei; Wang, Xiao

    2014-01-01

    Silage making has become a significant method of forage conservation worldwide. To determine how tomato pomace (TP) may be used effectively as animal feed, it was ensilaged for 90 days and microbiology counts, fermentation characteristics and chemical composition of tomato pomace silage (TPS) were evaluated at the 30th, 60th, and 90th days, respectively. In addition, 103 lactic acid bacteria were isolated from TPS. Based on the phenotypic and chemotaxonomic characteristics, 16S rDNA sequence and carbohydrate fermentation tests, the isolates were identified as 17 species namely: Lactobacillus coryniformis subsp. torquens (0.97%), Lactobacillus pontis (0.97%), Lactobacillus hilgardii (0.97%), Lactobacillus pantheris (0.97%), Lactobacillus amylovorus (1.9%), Lactobacillus panis (1.9%), Lactobacillus vaginalis (1.9%), Lactobacillus rapi (1.9%), Lactobacillus buchneri (2.9%), Lactobacillus parafarraginis (2.9%), Lactobacillus helveticus (3.9%), Lactobacillus camelliae (3.9%), Lactobacillus fermentum (5.8%), Lactobacillus manihotivorans (6.8%), Lactobacillus plantarum (10.7%), Lactobacillus harbinensis (16.5%) and Lactobacillus paracasei subsp. paracasei (35.0%). This study has shown that TP can be well preserved for 90 days by ensilaging and that TPS is not only rich in essential nutrients, but that physiological and biochemical properties of the isolates could provide a platform for future design of lactic acid bacteria (LAB) inoculants aimed at improving the fermentation quality of silage. PMID:25049999

  8. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Delyan P Ivanov

    Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  9. Isolation and characteristics of lactic acid bacteria isolated from ripe mulberries in Taiwan

    Yi-sheng Chen

    2010-12-01

    Full Text Available The objective of this study was to isolate, characterize, and identify lactic acid bacteria (LAB from ripe mulberries collected in Taiwan. Ripe mulberry samples were collected at five mulberry farms, located in different counties of Taiwan. Eighty-eight acid-producing cultures were isolated from these samples, and isolates were divided into classes first by phenotype, then into groups by restriction fragment length polymorphism (RFLP analysis and sequencing of 16S ribosomal DNA (rDNA. Phenotypic and biochemical characteristics led to identification of four bacterial groups (A to D. Weissella cibaria was the most abundant type of LAB distributed in four mulberry farms, and Lactobacillus plantarum was the most abundant LAB found in the remaining farm. Ten W. cibaria and one Lactococcus lactis subsp. lactis isolate produced bacteriocins against the indicator strain Lactobacillus sakei JCM 1157T. These results suggest that various LAB are distributed in ripe mulberries and W. cibaria was the most abundant LAB found in this study.

  10. Simple method of isolating humic acids from organic soils

    Ahmed, O. H.; Susilawati, K.; Nik Muhamad, A. B.; Khanif, M. Y.

    2009-04-01

    Humic substances particularly humic acids (HA) play a major role in soil conditioning e.g. erosion control, soil cation exchange capacity, complexation of heavy metal ions and pesticides, carbon and nitrogen cycles, plant growth and reduction of ammonia volatilization from urea. Humified substances such as coal, composts, and peat soils have substantial amounts of HA but the isolation of these acids is expensive, laborious, and time consuming. Factors that affect the quality and yield of HA isolated from these materials include extraction, fractionation, and purification periods. This work developed a simple, rapid, and cost effective method of isolating HA from peat soils. There was a quadratic relationship between extraction period and HA yield. Optimum extraction period was estimated at 4 h instead of the usual range of 12 to 48 h. There was no relationship between fractionation period and HA yield. As such 2 h instead of the usual range of 12 to 24 h fractionation period could be considered optimum. Low ash content (5%), remarkable reduction in K, coupled with the fact that organic C, E4/E6, carboxylic COOH, phenolic OH, and total acidity values of the HA were consistent with those reported by other authors suggest that the HA dealt with were free from mineral matter. This was possible because the distilled water used to purify the HA served as Bronsted-Lowry acid during the purification process of the HA. Optimum purification period using distilled waster was 1 h instead of the usual range of 1 and 7 days (uses HF and HCl and dialysis). Humic acids could be isolated from tropical peat soils within 7 h (i.e. 4 h extraction, 2 h fractionation, and 1 h purification) instead of the existing period of 2 and 7 days. This could facilitate the idea of producing organic fertilizers such as ammonium-humate and potassium-humate from humified substances since techniques devised in this study did not alter the true nature of the HA. Besides, the technique is rapid, simple

  11. Lactic acid bacteria isolated from soy sauce mash in Thailand.

    Tanasupawat, Somboon; Thongsanit, Jaruwan; Okada, Sanae; Komagata, Kazuo

    2002-08-01

    Fourteen sphere-shaped and 30 rod-shaped lactic acid bacteria were isolated from soy sauce mash of two factories in Thailand. These strains were separated into two groups, Group A and Group B, by cell shape and DNA-DNA similarity. Group A contained 14 tetrad-forming strains, and these strains were identified as Tetragenococcus halophilus by DNA similarity. Group B contained 30 rod-shaped bacteria, and they were further divided into four Subgroups, B1, B2, B3, and B4, and three ungrouped strains by phenotypic characteristics and DNA similarity. Subgroup B1 contained 16 strains, and these strains were identified as Lactobacillus acidipiscis by DNA similarity. Subgroup B2 included two strains, and the strains were identified as Lactobacillus farciminis by DNA similarity. Subgroup B3 contained five strains. The strains had meso-diaminopimelic acid in the cell wall, and were identified as Lactobacillus pentosus by DNA similarity. The strains tested produced DL-lactic acid from D-glucose. Subgroup B4 contained four strains. The strains had meso-diaminopimelic acid in the cell wall, and they were identified as Lactobacillus plantarum by DNA similarity. Two ungrouped strains were homofermentative, and one was heterofermentative. They showed a low degree of DNA similarity with the type strains tested, and were left unnamed. The distribution of lactic acid bacteria in soy sauce mash in Thailand is discussed. PMID:12469319

  12. O- and N-glycosylation of the Leishmania mexicana-secreted acid phosphatase. Characterization of a new class of phosphoserine-linked glycans.

    Ilg, T; Overath, P; Ferguson, M A; Rutherford, T; Campbell, D G; McConville, M J

    1994-09-30

    The protozoan parasite Leishmania mexicana secretes a heavily glycosylated 100-kDa acid phosphatase (sAP) which is associated with one or more polydisperse proteophosphoglycans. Most of the glycans in this complex were released using mild acid hydrolysis conditions that preferentially cleave phosphodiester linkages. The released saccharides were shown to consist of monomeric mannose and a series of neutral and phosphorylated glycans by Dionex high performance liquid chromatography, methylation analysis, exoglycosidase digestions, and one-dimensional 1H NMR spectroscopy. The neutral species comprised a linear series of oligosaccharides with the structures [Man alpha 1-2]1-5Man. The phosphorylated oligosaccharides were characterized as PO4-6Gal beta 1-4Man and PO4-6[Glc beta 1-3]Gal beta 1-4Man. The attachment of these glycans to the polypeptide backbone via the linkage, Man alpha 1-PO4-Ser, is suggested by: 1) the finding that more than 60% of the serine residues in the polypeptide are phosphorylated and 2) the resistance of the phosphoserine residues to alkaline phosphatase digestion unless the sAP was first treated with either mild acid (to release all glycans) or jack bean alpha-mannosidase (to release neutral mannose glycans). Analysis of the partially resolved components of the complex indicated that the most of the O-linked glycans on the 100-kDa phosphoglycoprotein comprised mannose and the mannose-oligosaccharides. In contrast the major O-linked glycans on the proteophosphoglycan were short phosphoglycan chains, containing on average two repeat units per chain. In addition to the O-linked glycans, both components in the sAP complex contained N-linked glycans. The N-glycanase F-released glycans were characterized by Bio-Gel P4 chromatography and exoglycosidase digestions to be the biantennary oligomannose type with the structures Glc1Man6GlcNAc2 and Man6GlcNAc2. The O-linked glycans of the sAP complex are similar to those found in the phosphoglycan chains of

  13. ALP (Alkaline Phosphatase) Test

    ... Also known as: ALK PHOS; Alkp Formal name: Alkaline Phosphatase Related tests: AST ; ALT ; GGT ; Bilirubin ; Liver Panel ; Bone Markers ; Alkaline Phosphatase Isoenzymes; Bone Specific ALP All content on ...

  14. Ethylene signalling is involved in regulation of phosphate starvation-induced gene expression and production of acid phosphatases and anthocyanin in Arabidopsis

    Lei, Mingguang

    2010-11-30

    With the exception of root hair development, the role of the phytohormone ethylene is not clear in other aspects of plant responses to inorganic phosphate (Pi) starvation. The induction of AtPT2 was used as a marker to find novel signalling components involved in plant responses to Pi starvation. Using genetic and chemical approaches, we examined the role of ethylene in the regulation of plant responses to Pi starvation. hps2, an Arabidopsis mutant with enhanced sensitivity to Pi starvation, was identified and found to be a new allele of CTR1 that is a key negative regulator of ethylene responses. 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, increases plant sensitivity to Pi starvation, whereas the ethylene perception inhibitor Ag+ suppresses this response. The Pi starvation-induced gene expression and acid phosphatase activity are also enhanced in the hps2 mutant, but suppressed in the ethylene-insensitive mutant ein2-5. By contrast, we found that ethylene signalling plays a negative role in Pi starvation-induced anthocyanin production. These findings extend the roles of ethylene in the regulation of plant responses to Pi starvation and will help us to gain a better understanding of the molecular mechanism underlying these responses. © 2010 The Authors. New Phytologist © 2010 New Phytologist Trust.

  15. Cell- and ligand-specific dephosphorylation of acid hydrolases: Evidence that the mannose 6-phosphatase is controlled by compartmentalization

    Einstein, R.; Gabel, C.A. (Columbia Univ. College of Physicians and Surgeons, NY (USA))

    1991-01-01

    Mouse L cells that possess the cation-independent mannose 6-phosphate (Man 6-P)/insulin-like growth factor (IGF) II receptor change the extent to which they dephosphorylate endocytosed acid hydrolases in response to serum. To investigate the mechanism by which dephosphorylation competence is regulated, the dephosphorylation of individual acid hydrolases was studied in Man 6-P/IGF II receptor-positive and -deficient cell lines. 125I-labeled Man 6-P-containing acid hydrolases were proteolytically processed but remained phosphorylated when endocytosed by receptor-positive L cells maintained in the absence of serum; after the addition of serum, however, the cell-associated hydrolases were dephosphorylated. Individual hydrolases were dephosphorylated at distinct rates and to different extents. In contrast, the same hydrolases were dephosphorylated equally and completely after entry into Man 6-P/IGF II receptor-positive Chinese hamster ovary (CHO) cells. The dephosphorylation competence of Man 6-P/IGF II receptor-deficient mouse J774 cells was more limited. beta-Glucuronidase produced by these cells underwent a limited dephosphorylation in transit to lysosomes such that diphosphorylated oligosaccharides were converted to monophosphorylated species. The overall quantity of phosphorylated oligosaccharides associated with the enzyme, however, did not decrease within the lysosomal compartment. Likewise, beta-glucuronidase was not dephosphorylated when introduced into J774 cells via Fc receptor-mediated endocytosis. The CHO and J774 cell lysosomes, therefore, display opposite extremes with respect to their capacity to dephosphorylate acid hydrolases; within CHO cell lysosomes acid hydrolases are rapidly and efficiently dephosphorylated, but within J774 cell lysosomes the same acid hydrolases remain phosphorylated.

  16. Cell- and ligand-specific dephosphorylation of acid hydrolases: Evidence that the mannose 6-phosphatase is controlled by compartmentalization

    Mouse L cells that possess the cation-independent mannose 6-phosphate (Man 6-P)/insulin-like growth factor (IGF) II receptor change the extent to which they dephosphorylate endocytosed acid hydrolases in response to serum. To investigate the mechanism by which dephosphorylation competence is regulated, the dephosphorylation of individual acid hydrolases was studied in Man 6-P/IGF II receptor-positive and -deficient cell lines. 125I-labeled Man 6-P-containing acid hydrolases were proteolytically processed but remained phosphorylated when endocytosed by receptor-positive L cells maintained in the absence of serum; after the addition of serum, however, the cell-associated hydrolases were dephosphorylated. Individual hydrolases were dephosphorylated at distinct rates and to different extents. In contrast, the same hydrolases were dephosphorylated equally and completely after entry into Man 6-P/IGF II receptor-positive Chinese hamster ovary (CHO) cells. The dephosphorylation competence of Man 6-P/IGF II receptor-deficient mouse J774 cells was more limited. beta-Glucuronidase produced by these cells underwent a limited dephosphorylation in transit to lysosomes such that diphosphorylated oligosaccharides were converted to monophosphorylated species. The overall quantity of phosphorylated oligosaccharides associated with the enzyme, however, did not decrease within the lysosomal compartment. Likewise, beta-glucuronidase was not dephosphorylated when introduced into J774 cells via Fc receptor-mediated endocytosis. The CHO and J774 cell lysosomes, therefore, display opposite extremes with respect to their capacity to dephosphorylate acid hydrolases; within CHO cell lysosomes acid hydrolases are rapidly and efficiently dephosphorylated, but within J774 cell lysosomes the same acid hydrolases remain phosphorylated

  17. Isolation of lactic acid bacteria for its possible use in the fermentation of green algerian olives

    Nour-Eddine, Karam; Halima, Zadi-Karam; Mourad, Kacem

    2004-01-01

    This study was undertaken with the aim of obtaining lactic acid bacteria with the ability to ferment olives for possible use as starter cultures. For this reason, 32 isolates of lactic acid bacteria isolated from the spontaneous fermentation of green olives were characterized and identified on the basis of morphological and biochemical criteria. 14 of them were identified as Lactococcus lactis, 11 isolates as Lactobacillus plantarum and 7 isolates as Enterococcus sp. Of the 18 isolates examin...

  18. The phosphatase activity of the isolated H4-H5 loop of Na+/K+ ATPase resides outside its ATP binding site

    Ettrich, Rüdiger

    2004-01-01

    Roč. 271, č. 19 (2004), s. 3923-3939. ISSN 0014-2956 Institutional research plan: CEZ:AV0Z5011922; MSM113100003 Keywords : phosphatase Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.260, year: 2004

  19. Improvement of Student Understanding of How Kinetic Data Facilitates the Determination of Amino Acid Catalytic Function through an Alkaline Phosphatase Structure/Mechanism Bioinformatics Exercise

    Grunwald, Sandra K.; Krueger, Katherine J.

    2008-01-01

    Laboratory exercises, which utilize alkaline phosphatase as a model enzyme, have been developed and used extensively in undergraduate biochemistry courses to illustrate enzyme steady-state kinetics. A bioinformatics laboratory exercise for the biochemistry laboratory, which complements the traditional alkaline phosphatase kinetics exercise, was…

  20. Isolation and Identification of Lactic Acid Bacteria Isolated from a Traditional Jeotgal Product in Korea

    Cho, Gyu Sung; Do, Hyung Ki

    2006-06-01

    Seventeen lactic acid bacterial strains (LAB) were isolated using MRS agar medium from Jeotgal, a Korean fermented food, purchased at the Jukdo market of Pohang. To identify the strains isolated, they were tested by examining their cell morphologies, gram-staining, catalase activity, arginine hydrolase activity, D-L lactate form and carbohydrate fermentation. According to the phenotypic characteristics, three strains were tent atively identified as Lactobacillus spp., ten were Enterococcus spp. (or Streptococcus spp., or Pediococcus spp.) and the rest were Leuconostoc spp. (or Weissella spp.). Five strains among 17 were chosen by preliminary bacteriocin activity test. Four bacterial strains which inhibited both indicator microorganisms were identified by 16S rRNA sequencing. The results are as follows; Leuconostoc mesenteroides (HK 4), Leuconostoc mesenteroides (HK 5), Leuconostoc mesenteroides(HK 11), Streptococcus salivarius(HK 8). In order to check LAB which are showing a high survival rate in gut, we investigated three strains inhibiting both indicator microorganisms in artificial gastric acid and bile juice -all except HK8. The three strains mentioned above grew in extreme low acid conditions.

  1. Flotation isolation of uranium(6) from acidic waste waters with the use of potassium soaps of dialkylphosphinic acids

    The applicability of potassium soaps of dialkylphosphinic acids of R2POOH type where R is C3-C10 hydrocarbon radical as uranium flotation collectors is investigated. Under similar conditions the degree of uranium flotation isolation from nitrate solutions is notably higher than from chloride solutions, and from the latter is higher than from sulfate solutions. The method of uranium flotation isolation using dialkylphosphinic acid soaps was tested in synthetic solutions simulating acid (pH = 2-5) waste waters. Possibility of flotation method to isolate 95-99% of uranium from acid solutions using potassium soaps of dialkylphosphinic acids is demonstrated

  2. Isolation and seasonal effects on characteristics of fulvic acid isolated from an Australian floodplain river and billabong.

    McDonald, Suzanne; Pringle, Jennifer M; Bishop, Andrea G; Prenzler, Paul D; Robards, Kevin

    2007-06-15

    Fulvic acids from an Australian floodplain river and billabong were isolated using DEAE and DAX-8 resins, and characterised with the use of size exclusion chromatography and solid-state CP-MAS (13)C NMR spectroscopy. Differences between the two resin isolates were evident. Fulvic acids isolated using DEAE-cellulose had higher apparent M(n) and M(w) values, while the DAX-8 resin showed a slight preference for aliphatic components. Fulvic acids from the river and billabong had the same functional groups present, however, the river fulvic acids had higher apparent M(n) (number average molecular weight) and M(w) values (weight average molecular weight), and were more polydisperse than the billabong fulvic acid. There were no significant changes in the characteristics of the fulvic acid isolated from the river at four sampling times: summer, autumn, winter and spring. In contrast, fulvic acids isolated from a billabong displayed seasonal variation in molecular weights. This work emphasizes the importance in ecological studies of isolation procedure for the operationally defined fulvic acids. PMID:17010354

  3. Comparison of the effects of eldecalcitol with either raloxifene or bisphosphonate on serum tartrate resistant acid phosphatase-5b, a bone resorption marker, in postmenopausal osteoporosis

    Takada, Junichi; Ikeda, Satoshi; Kusanagi, Tetsuya; Mizuno, Satoshi; Wada, Hiroshi; Iba, Kousuke; Yoshizaki, Takashi; Yamashita, Toshihiko

    2016-01-01

    Summary Objective This study analyzes whether concomitant raloxifene (RLX) or bisphosphonates (BP) plus eldecalcitol (ELD) has excessive suppressive effects on a bone resorption marker during the first 6 months of treatment in postmenopausal women in real-world setting. Methods 285 postmenopausal osteoporotic patients who had been treated with RLX or BP plus ELD were evaluated the bone resorption marker, serum tartrate resistant acid phosphatase-5b (TRACP-5b), during the first 6 months of treatment. Results In drug-naïve group (not received osteoporosis medications before the administration, n=70), the concomitant RLX or BP with ELD significantly decreased levels of TRACP-5b without severe suppression. In vitamin D switch group [RLX or BP plus alfacalcidol (ALF) and then switched to RLX or BP plus ELD, n=215], the replacing ALF with ELD further and significantly decreased TRACP-5b and tertile analyses based on baseline values were significantly decreased far more in the highest, compared with the lowest tertile in the ELD+RLX and ELD+BP groups. Conclusion ELD combined with RLX or BP administered for 6 months to postmenopausal women with osteoporosis who were drug-naïve or who had switched medications significantly reduced and maintained TRACP-5b values within the reference range. PMID:27252739

  4. Effect of colchicine on the Golgi apparatus and on GERL of rat jejunal absorptive cells. Ultrastructural localization of thiamine pyrophosphatase and acid phosphatase activity.

    Pavelka, M; Ellinger, A

    1981-04-01

    Ultrastructural localization of thiamine pyrophosphatase (TTP) and acid phosphatase (AcPase) activity was performed on jejunal absorptive cells of rats pretreated with the antimicrotubular agent colchicine and of control animals. Demonstration of TPP activity showed that most of the dislocated Golgi stacks after colchicine application lacked positively staining cisternae of the mature side. This cytochemical finding is in agreement with the morphologically demonstrable changes of the Golgi stacks resulting in a loss of polarity and give evidence for a colchicine-induced deficiency of the Golgi apparatus. The cytochemical localization of AcPase activity showed deposits of reaction product over lysosomes and GERL and demonstrated a dislocation of GERL occurring concomitantly with the changes of the Golgi apparatus. The antimicrotubular effect of colchicine is well documented; thus the morphological and cytochemical changes of the Golgi apparatus and of GERL might be due to a disturbed microtubular function after application of this agent suggesting an influence of microtubules in the maintenance of the integrity of these organelles. This hypothesis includes the possibility of an involvement of microtubules in formation and differentiation of Golgi stacks and GERL as well as a kind of "skeletal"function being responsible for their characteristic structure and fashion. PMID:6113143

  5. The spatial distribution of acid phosphatase activity in ectomycorrhizal tissues depends on soil fertility and morphotype, and relates to host plant phosphorus uptake.

    Alvarez, Maricel; Huygens, Dries; Díaz, Leila Milena; Villanueva, Claudia Añazco; Heyser, Wolfgang; Boeckx, Pascal

    2012-01-01

    Acid phosphatase (ACP) enzymes are involved in the mobilization of soil phosphorus (P) and polyphosphate accumulated in the fungal tissues of ectomycorrhizal roots, thereby influencing the amounts of P that are stored in the fungus and transferred to the host plant. This study evaluated the effects of ectomycorrhizal morphotype and soil fertility on ACP activity in the extraradical mycelium (ACP(myc)), the mantle (ACP(mantle)) and the Hartig net region (ACP(Hartig)) of ectomycorrhizal Nothofagus obliqua seedlings. ACP activity was quantified in vivo using enzyme-labelled fluorescence-97 (ELF-97) substrate, confocal laser microscopy and digital image processing routines. There was a significant effect of ectomycorrhizal morphotype on ACP(myc), ACP(mantle) and ACP(Hartig), while soil fertility had a significant effect on ACP(myc) and ACP(Hartig). The relative contribution of the mantle and the Hartig net region to the ACP activity on the ectomycorrhizal root was significantly affected by ectomycorrhizal morphotype and soil fertility. A positive correlation between ACP(Hartig) and the shoot P concentration was found, providing evidence that ACP activity at the fungus:root interface is involved in P transfer from the fungus to the host. It is concluded that the spatial distribution of ACP in ectomycorrhizas varies as a function of soil fertility and colonizing fungus. PMID:21902696

  6. Biochemical effect of a histidine phosphatase acid (phytase) of Aspergillus japonicus var. Saito on performance and bony characteristics of broiler.

    Maller, Alexandre; de Quadros, Thays Cristina Oliveira; Junqueira, Otto M; Graña, Alfredo Lora; de Lima Montaldi, Ana Paula; Alarcon, Ricardo Fernandes; Jorge, João Atílio; de Lourdes T M Polizeli, Maria

    2016-01-01

    Phytases are enzymes that hydrolyze the ester linkage of phytic acid, releasing inositol and inorganic phosphate. The phytic acid (phytate) is a major form of phosphorus in plant foods. Knowing that diet for animal of production has the cereal base (corn and soybean), primarily, broilers need for an alternative to use of the phosphate present in these ingredients, since it does not naturally produce the enzyme phytase, which makes it available. The aims of this work was studding the safe supplementation of Aspergillus japonicus var. Saito crude phytase in feeding broilers and check the biochemical effect on performance and bones of these animals. The enzymatic extract did not have aflatoxins B1, B2, G2 and G1 and zearalenone and ochratoxin, and low concentrations of this extract did not have cytotoxic effects on cells derived from lung tissue. The in vivo experiments showed that the phytase supplied the available phosphate reduction in the broiler feed formulation, with a live weight, weight gain, feed intake, feed conversion, viability, productive efficiency index and carcass yield similar to the control test. Furthermore, the phytase supplementation favored the formation of bone structure and performance of the broilers. The results show the high biotechnological potential of A. japonicus phytase on broiler food supplementation to reduce phosphorus addition in the food formulation. So, this enzyme could be used as a commercial alternative to animal diet supplementation. PMID:27625972

  7. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    The overall goal of this project is to examine the role of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium [U(VI)] phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO43-. During this phase of the project we have been conducting assays to determine the effects of pH, inorganic anions and organic ligands on U(VI) mineral formation and precipitation when FRC bacterial isolates were grown in simulated groundwater medium. The molecular characterization of FRC isolates has also been undertaken during this phase of the project. Analysis of a subset of gram-positive FRC isolates cultured from FRC soils (Areas 1, 2 and 3) and background sediments have indicated a higher percentage of isolates exhibiting phosphatase phenotypes (i.e., in particular those surmised to be PO43--irrepressible) relative to isolates from the reference site. A high percentage of strains that exhibited such putatively PO43--irrepressible phosphatase phenotypes were also resistant to the heavy metals lead and cadmium. Previous work on FRC strains, including Arthrobacter, Bacillus and Rahnella spp., has demonstrated differences in tolerance to U(VI) toxicity (200 (micro)M) in the absence of organophosphate substrates. For example, Arthrobacter spp. exhibited the greatest tolerance to U(VI) while the Rahnella spp. have been shown to facilitate the precipitation of U(VI) from solution and the Bacillus spp. demonstrate the greatest sensitivity to acidic conditions and high concentrations of U(VI). PCR-based detection of FRC strains are being conducted to determine if non-specific acid phosphatases of the known molecular classes [i.e., classes A, B and C] are present in these FRC isolates. Additionally, these amplified phosphatases are being

  8. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    Martinez, Robert J.; Beazley, Melanie J.; Wilson, Jarad J.; Taillefert, Martial; Sobecky, Patricia A.

    2005-04-05

    The overall goal of this project is to examine the role of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium [U(VI)] phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO{sub 4}{sup 3-}. During this phase of the project we have been conducting assays to determine the effects of pH, inorganic anions and organic ligands on U(VI) mineral formation and precipitation when FRC bacterial isolates were grown in simulated groundwater medium. The molecular characterization of FRC isolates has also been undertaken during this phase of the project. Analysis of a subset of gram-positive FRC isolates cultured from FRC soils (Areas 1, 2 and 3) and background sediments have indicated a higher percentage of isolates exhibiting phosphatase phenotypes (i.e., in particular those surmised to be PO{sub 4}{sup 3-}-irrepressible) relative to isolates from the reference site. A high percentage of strains that exhibited such putatively PO{sub 4}{sup 3-}-irrepressible phosphatase phenotypes were also resistant to the heavy metals lead and cadmium. Previous work on FRC strains, including Arthrobacter, Bacillus and Rahnella spp., has demonstrated differences in tolerance to U(VI) toxicity (200 {micro}M) in the absence of organophosphate substrates. For example, Arthrobacter spp. exhibited the greatest tolerance to U(VI) while the Rahnella spp. have been shown to facilitate the precipitation of U(VI) from solution and the Bacillus spp. demonstrate the greatest sensitivity to acidic conditions and high concentrations of U(VI). PCR-based detection of FRC strains are being conducted to determine if non-specific acid phosphatases of the known molecular classes [i.e., classes A, B and C] are present in these FRC isolates. Additionally, these

  9. AtPP2CG1, a protein phosphatase 2C, positively regulates salt tolerance of Arabidopsis in abscisic acid-dependent manner

    Liu, Xin, E-mail: fangfei6073@126.com [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Zhu, Yanming, E-mail: ymzhu2001@neau.edu.cn [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Zhai, Hong, E-mail: Zhai.h@neigaehrb.ac.cn [Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin 150040 (China); Cai, Hua, E-mail: small-big@sohu.com [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Ji, Wei, E-mail: iwei_j@hotmail.com [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Luo, Xiao, E-mail: luoxiao2010@yahoo.cn [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China); Li, Jing, E-mail: lijing@neau.edu.cn [Plant Secondary Metabolism Laboratory, Northeast Agricultural University, Harbin 150030 (China); Bai, Xi, E-mail: baixi@neau.edu.cn [Plant Bioengineering Laboratory, Northeast Agricultural University, Harbin 150030 (China)

    2012-06-15

    Highlights: Black-Right-Pointing-Pointer AtPP2CG1 positively regulates salt tolerance in ABA-dependent manner. Black-Right-Pointing-Pointer AtPP2CG1 up-regulates the expression of marker genes in different pathways. Black-Right-Pointing-Pointer AtPP2CG1 expresses in the vascular system and trichomes of Arabidopsis. -- Abstract: AtPP2CG1 (Arabidopsis thaliana protein phosphatase 2C G Group 1) was predicted as an abiotic stress candidate gene by bioinformatic analysis in our previous study. The gene encodes a putative protein phosphatase 2C that belongs to Group G of PP2C. There is no report of Group G genes involved in abiotic stress so far. Real-time RT-PCR analysis showed that AtPP2CG1 expression was induced by salt, drought, and abscisic acid (ABA) treatment. The expression levels of AtPP2CG1 in the ABA synthesis-deficient mutant abi2-3 were much lower than that in WT plants under salt stress suggesting that the expression of AtPP2CG1 acts in an ABA-dependent manner. Over-expression of AtPP2CG1 led to enhanced salt tolerance, whereas its loss of function caused decreased salt tolerance. These results indicate that AtPP2CG1 positively regulates salt stress in an ABA-dependent manner. Under salt treatment, AtPP2CG1 up-regulated the expression levels of stress-responsive genes, including RD29A, RD29B, DREB2A and KIN1. GUS activity was detected in roots, leaves, stems, flower, and trichomes of AtPP2CG1 promoter-GUS transgenic plants. AtPP2CG1 protein was localized in nucleus and cytoplasm via AtPP2CG1:eGFP and YFP:AtPP2CG1 fusion approaches.

  10. The small chemical enzyme inhibitor 5-phenylnicotinic acid/CD13 inhibits cell migration and invasion of tartrate-resistant acid phosphatase/ACP5-overexpressing MDA-MB-231 breast cancer cells.

    Krumpel, Michael; Reithmeier, Anja; Senge, Teresa; Baeumler, Toni Andreas; Frank, Martin; Nyholm, Per-Georg; Ek-Rylander, Barbro; Andersson, Göran

    2015-11-15

    Tartrate-resistant acid phosphatase (TRAP/ACP5/uteroferrin/purple acid phosphatase/PP5) has received considerable attention as a newly discovered proinvasion metastasis driver associated with different malignancies. This renders TRAP an interesting target for novel anti-cancer therapy approaches. TRAP exists as two isoforms, 5a and 5b, where the 5a isoform represents an enzymatically less active monomeric precursor to the more enzymatically active 5b isoform generated by proteolytic excision of a repressive loop domain. Recently, three novel lead compounds were identified by fragment-based screening and demonstrated to be efficient TRAP enzyme inhibitors in vitro. We conclude that one of the three compounds i.e. 5-phenylnicotinic acid (CD13) was efficient as a TRAP inhibitor with Kic values in the low micromolar range towards the TRAP 5b isoform, but was not able to inhibit the TRAP 5a isoform. Structure-based docking revealed similar interactions of CD13 with the active site in both TRAP isoforms. In stably TRAP-overexpressing MDA-MB-231 breast cancer cells, CD13 inhibited intracellular TRAP activity and showed no cytotoxicity at 200 µM. Furthermore, CD13 selectively blocked the TRAP 5b isoform compared to the TRAP 5a in cultured cells, indicating the usefulness of CD13 for assessing the different biological functions of the two TRAP isoforms 5a and 5b in cell systems. Moreover, inhibition of cell migration and invasion of stably TRAP-overexpressing MDA-MB-231 by CD13 was observed. These data establish a proof of principle that a small chemical inhibitor of the TRAP enzyme can block TRAP-dependent functions in cancer cells. PMID:26428664

  11. Isolation, enumeration, molecular identification and probiotic potential evaluation of lactic acid bacteria isolated from sheep milk

    L.B. Acurcio

    2014-06-01

    Full Text Available Lactic acid bacteria species were molecularly identified in milk from Lacaune, Santa Inês and crossbred sheep breeds and their in vitro probiotic potential was evaluated. The species identified were Enterococcus faecium (56.25%, E. durans (31.25% and E. casseliflavus (12.5%. No other lactic acid bacteria species, such as lactobacilli, was identified. Most of the isolated enterococci were resistant to gastric pH (2.0 and to 0.3% oxgall. All tested enterococci were resistant to ceftazidime, oxacillin and streptomycin and sensible to clindamycin, erythromycin and penicillin. The resistance to ciprofloxacin, gentamicin, tetracycline and vancomycin varied among tested species. All tested enterococci strongly inhibited (P<0.05 Escherichia coli and Listeria monocytogenes, moderately inhibited E. faecalis and Staphylococcus aureus and did not inhibit Pseudomonas aeruginosa, Salmonella enterica var. Typhimurium and also one E. durans sample isolated from sheep milk. Four samples of E. faecium, one of E. durans and one of E. casseliflavus presented the best probiotic potential.

  12. ISOLATION OF PECTIC ACIDS FROM BLEACHED TMP WATER AND AGGREGATION OF MODEL AND TMP PECTIC ACIDS BY CALCIUM

    Anna C. Sundberg

    2007-11-01

    Full Text Available Pectins are important structural elements in spruce fibres. Alkaline peroxide bleaching of spruce thermomechanical pulp (TMP causes degradation and demethylation of pectins, yielding high-charge-density pectic acids. The pectic acids in fibres contribute strongly to the negative fibre charge, and the dissolved pectic acids increase the cationic demand of bleached TMP water. In this study, a method to isolate pectic acids from peroxide-bleached TMP pulp water is presented. The pectic acids were isolated and purified in good yield using a polyacrylate resin to remove lignin, a cellulose filter to remove galactoglucomannans (GGM, and an anion exchange resin to separate pectic acids from neutral carbohydrates. Salts and residual low-molar-mass carbohydrates were further removed from the isolated pectic acids by dialysis. The isolated pectic acids (>80% purity had a low molar mass and a wide polydispersity (5.9 kDa, MW/MN 3.3. The aggregation and precipitation of the isolated pectic acids, as well as citrus fruit pectic acids with well-defined molar masses, by Ca2+-ions were studied. The molar mass of pectic acids was a key factor determining the precipitation of Ca2+-pectates. Pectic acids below 6 kDa were not precipitated by Ca2+, while higher molar masses led first to partial and then to complete precipitation. The precipitated Ca2+-pectates may impair paper machine runnability and paper quality.

  13. Tartrate-resistant acid phosphatase (TRAP) co-localizes with receptor activator of NF-KB ligand (RANKL) and osteoprotegerin (OPG) in lysosomal-associated membrane protein 1 (LAMP1)-positive vesicles in rat osteoblasts and osteocytes

    Solberg, L. B.; Stang, E.; Brorson, S.-H.; Andersson, G; Reinholt, F.P.

    2014-01-01

    Tartrate-resistant acid phosphatase (TRAP) is well known as an osteoclast marker; however, a recent study from our group demonstrated enhanced number of TRAP + osteocytes as well as enhanced levels of TRAP located to intracellular vesicles in osteoblasts and osteocytes in experimental osteoporosis in rats. Such vesicles were especially abundant in osteoblasts and osteocytes in cancellous bone as well as close to bone surface and intracortical remodeling sites. To further investigate TRAP in o...

  14. Isolation and identification of indigenous lactic acid bacteria from North Sumatra river buffalo milk

    2015-01-01

    Buffalo milk is a source of various lactic acid bacteria (LAB) which is potential as culture starter as well as the probiotic. This study was conducted to isolate and identify LAB from indigenous North Sumatra river buffalo milk. Lactic acid bacteria was isolated and grown in medium De Man Rogosa Sharpe Agar (MRSA). The isolation was conducted to obtain pure isolate. The identification of LAB was studied in terms of morphology, physiology, biochemistry and survival on low pH. Morphology test...

  15. Bile Salt and Acid Tolerant of Lactic Acid Bacteria Isolated from Proventriculus of Broiler Chicken

    E. Damayanti

    2014-08-01

    Full Text Available The aim of this research was to obtain the lactic acid bacteria (LAB as probiotic candidates which have resistance to bile salt and acid condition. LAB was obtained using isolation method from proventriculus of broiler chicken. Selective MRS media with 0.2% CaCO3 addition were used for LAB isolation using pour plate sampling method under anaerobic condition. The result showed that four selected isolates had morphological and biochemical characteristics as LAB. The selected LAB was characterized as follow: antibacterial activities, antibiotic sensitivity, resistance on bile salt, gastric juice and acid condition, and biochemical identification. Antibacterial activities assay of cell free supernatant was confirmed using disc paper diffusion method which was arranged on factorial design and each treatment consisted of three replications. The cell free supernatant of LAB isolates had antibacterial activities against Escherichia coli, Pseudomonas aerugenosa, and Salmonella pullorum. Molecular identification procedure using 16S rRNA sequence analysis showed that R01 and R02 as Pediococcus acidilactici. The viability of the two isolates were tested by acid pH (pH 1, 2, and 3, gastric juice pH 2, and bile salt condition for digestives tract simulation. The result showed that R01 and R02 had a high viability percentages at pH 1, 2, and 3 (95.45%, 99.49%, 104.01%, and 67.17%, 120.74%, 103.4%, respectively and at bile salt simulation for 1-2 hours (100.35%-102.71% and 100.02%-102.65%, respectively, but at gastric juice simulation for 1-2 hours, the P. acidilactici R01 had higher viability than P. acidilactici R02 (59.69%-76.53% versus 43.57%-40.69%, respectively. In the antibiotic sensitivity test for three antibiotics (i.e. erythromicin 15 µg, penicillin G 10 µg, and streptomycin 10 µg, the P. acidilactici R02 showed resistance to Streptomycin and Penicillin. It is concluded that P. acidilactici R01 and P. acidilactici R02 isolated from proventriculus

  16. Properties of nanocellulose isolated from corncob residue using sulfuric acid, formic acid, oxidative and mechanical methods.

    Liu, Chao; Li, Bin; Du, Haishun; Lv, Dong; Zhang, Yuedong; Yu, Guang; Mu, Xindong; Peng, Hui

    2016-10-20

    In this work, nanocellulose was extracted from bleached corncob residue (CCR), an underutilized lignocellulose waste from furfural industry, using four different methods (i.e. sulfuric acid hydrolysis, formic acid (FA) hydrolysis, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-mediated oxidation, and pulp refining, respectively). The self-assembled structure, morphology, dimension, crystallinity, chemical structure and thermal stability of prepared nanocellulose were investigated. FA hydrolysis produced longer cellulose nanocrystals (CNCs) than the one obtained by sulfuric acid hydrolysis, and resulted in high crystallinity and thermal stability due to its preferential degradation of amorphous cellulose and lignin. The cellulose nanofibrils (CNFs) with fine and individualized structure could be isolated by TEMPO-mediated oxidation. In comparison with other nanocellulose products, the intensive pulp refining led to the CNFs with the longest length and the thickest diameter. This comparative study can help to provide an insight into the utilization of CCR as a potential source for nanocellulose production. PMID:27474618

  17. Alicyclobacillus fodiniaquatilis sp. nov., isolated from acid mine water.

    Zhang, Bo; Wu, Yu-Fan; Song, Jin-Long; Huang, Zhong-Sheng; Wang, Bao-Jun; Liu, Shuang-Jiang; Jiang, Cheng-Ying

    2015-12-01

    Two novel, Gram-stain-variable, moderately thermophilic, acidophilic, rod-shaped, endospore-forming bacteria, G45-16T and G45-17, were isolated from acid mine water of Zijin copper mine in Fujian Province, China. Phylogenetic analysis of 16S rRNA gene sequences showed that they were closely related to Alicyclobacillus acidoterrestris ATCC 49025T with sequence similarities of 96.8 %. Cells grew aerobically at 20-45 °C (optimum, 40 °C), at pH 2.5-5.5(optimum, pH 3.5) and in the presence of 0-4.0 % (w/v) NaCl. Strains contained MK-7 as the major menaquinone and the major cellular fatty acids were ω-cyclohexane C19 : 0 and ω-cyclohexane C17 : 0. The DNA G+C content was 51.3 and 49.8 mol% (Tm) for G45-16T and G45-17, respectively. On the basis of phenotypic, chemotaxonomic and phylogenetic comparisons with their relatives and DNA-DNA relatedness values, it is concluded that strains G45-16T and G45-17 represent a novel species within the genus Alicyclobacillus, for which the name Alicyclobacillus fodiniaquatilis sp. nov. is proposed; the type strain is G45-16T(=CGMCC 1.15049T=NBRC 111483T). PMID:26476812

  18. ISOLATION OF PECTIC ACIDS FROM BLEACHED TMP WATER AND AGGREGATION OF MODEL AND TMP PECTIC ACIDS BY CALCIUM

    Anna C. Sundberg; Andrey V. Pranovich; Ville J. Saarimaa; Bjarne R. Holmbom

    2007-01-01

    Pectins are important structural elements in spruce fibres. Alkaline peroxide bleaching of spruce thermomechanical pulp (TMP) causes degradation and demethylation of pectins, yielding high-charge-density pectic acids. The pectic acids in fibres contribute strongly to the negative fibre charge, and the dissolved pectic acids increase the cationic demand of bleached TMP water. In this study, a method to isolate pectic acids from peroxide-bleached TMP pulp water is presented. The pectic acids we...

  19. Phosphatases in plants.

    Schweighofer, Alois; Meskiene, Irute

    2015-01-01

    Reversible protein phosphorylation is an essential posttranslational modification mechanism executed by opposing actions of protein phosphatases and protein kinases. About 1,000 predicted kinases in Arabidopsis thaliana kinome predominate the number of protein phosphatases, of which there are only ~150 members in Arabidopsis. Protein phosphatases were often referred to as "housekeeping" enzymes, which act to keep eukaryotic systems in balance by counteracting the activity of protein kinases. However, recent investigations reveal the crucial and specific regulatory functions of phosphatases in cell signaling. Phosphatases operate in a coordinated manner with the protein kinases, to execute their important function in determining the cellular response to a physiological stimulus. Closer examination has established high specificity of phosphatases in substrate recognition and important roles in plant signaling pathways, such as pathogen defense and stress regulation, light and hormonal signaling, cell cycle and differentiation, metabolism, and plant growth. In this minireview we provide a compact overview about Arabidopsis protein phosphatase families, as well as members of phosphoglucan and lipid phosphatases, and highlight the recent discoveries in phosphatase research. PMID:25930691

  20. [Phosphatase activity in Amoeba proteus at pH 9.0].

    Sopina, V A

    2007-01-01

    In the free-living amoeba Amoeba proteus (strain B), after PAAG disk-electrophoresis of the homogenate supernatant, at using 1-naphthyl phosphate as a substrate and pH 9.0, three forms of phosphatase activity were revealed; they were arbitrarily called "fast", "intermediate", and "slow" phosphatases. The fast phosphatase has been established to be a fraction of lysosomal acid phosphatase that preserves some low activity at alkaline pH. The question as to which particular class the intermediate phosphatase belongs to has remained unanswered: it can be both acid phosphatase and protein tyrosine phosphatase (PTP). Based on data of inhibitor analysis, large substrate specificity, results of experiments with reactivation by Zn ions after inactivation with EDTA, other than in the fast and intermediate phosphatases localization in the amoeba cell, it is concluded that only slow phosphatase can be classified as alkaline phosphatase (EC 3.1.3.1). PMID:17933343

  1. Porównanie aktywności peroksydazy, katalazy i fosfatazy kwaśnej w liściach pora w okresie intensywnego wzrostu i pod koniec wegetacji roślin w rożnych warunkach uprawy [Dynamics of the changes of peroxidase, catalase, and acid phosphatase activities in leeks under the influence of nitrogen fertilization and irrigation

    E. Gurgul; E. Kołota; D. Ściążko

    2015-01-01

    Results obtained in a field experiment showed a high influence of irrigation and nitrogen fertilization on the activity of peroxidase, while acid phosphatase activity showed only small differences. The peroxidase activity depended to a large degree on the leeks growth stage. Maximum peroxidase, catalase and - in some cases – acid phosphatase activity were found at nitrogen doses higher than optimal for the plant growth and yield.

  2. Porównanie aktywności peroksydazy, katalazy i fosfatazy kwaśnej w liściach pora w okresie intensywnego wzrostu i pod koniec wegetacji roślin w rożnych warunkach uprawy [Dynamics of the changes of peroxidase, catalase, and acid phosphatase activities in leeks under the influence of nitrogen fertilization and irrigation

    E. Gurgul

    2015-06-01

    Full Text Available Results obtained in a field experiment showed a high influence of irrigation and nitrogen fertilization on the activity of peroxidase, while acid phosphatase activity showed only small differences. The peroxidase activity depended to a large degree on the leeks growth stage. Maximum peroxidase, catalase and - in some cases – acid phosphatase activity were found at nitrogen doses higher than optimal for the plant growth and yield.

  3. Wpływ nawożenia mineralnego i nawadniania na aktyumość peroksydazy, katalazy i fosfatazy kwaśnej w dwóch fazach wzrostu kapusty i porów [The influence of mineral fertilization and irrigation on the activity of peroxidase, catalase and acid phosphatase of cabbage and leek in two stages of growth

    E. Gurgul; E. Kołota

    2015-01-01

    During the growth of plant, the very distinct increase of enzymatic activity of peroxidase and catalase was observed, but in case of acid phosphatase in smaller degree. An irrigation caused the decreasing of activity of all tested enzymes in both stages of cabbage growth. However, in case of leeks leaves sprinkling irrigation stimulated activity of catalase and acid phosphatase in both stages and peroxidase in the second stage of growth. The effectiveness of the mineral nutritive was differen...

  4. PROBIOTIC POTENTIALS AMONG LACTIC ACID BACTERIA ISOLATED FROM CURD

    Shruthy VV

    2011-02-01

    Full Text Available Curd is a commonly used fermented milk product in India since time immemorial. The scientific use of curd as a source of probiotic (good bacteria for health has not been much examined. The yougurt (curd containing probiotics is in Indian market and highly acclaimed. Therefore the status of curd as a source of probiotics is in question and requires scientific examination of its content, so the study was carried out. Probiotic potentials of two bacterial isolates from 20 different curd samples were identified as Lactobacillus spp. by the determination of morphological, cultural, physiological and biochemical characteristics, were investigated. The antibacterial potential against diarrhoegenic bacterial pathogens was also examined. The reference strain used was Lactobacillus acidophilus, MTCC 447. The percentage survivability of the strains at pH 3.5, was found to be satisfactory (>90%. Bile salt resistance (0.3% sodium thioglycollate was found to be between 80.41% and 83.2%. The pH decrease of the strains with time showed slow acidification activity. The lactic acid production of the strains ranges from 1.83 ± 0.12 to 3.93 ± 0.07 g. The strains were β-galactosidase producer and were resistant to principal antibiotics tested. But the absence of plasmids showed that they are intrinsically resistant or chromosome encoded. Strains showed maximum inhibition zone against V. cholerae O139 (13.67 ± 0.57 to 15.33 ± 0.57 mm in comparison to other diarrhoeagenic bacteria. Only 10% of the examined curd samples had probiotic bacteria. Isolated strains of Lactobacillus spp. showed satisfactory probiotic potentials in comparison with reference strains and with antibacterial activity against diarrhoeagenic pathogens and thus maybe useful in the management of diarrhoea and also in functional food industry.

  5. Structural and biochemical characterization of human PR70 in isolation and in complex with the scaffolding subunit of protein phosphatase 2A.

    Rebecca Dovega

    Full Text Available Protein Phosphatase 2A (PP2A is a major Ser/Thr phosphatase involved in the regulation of various cellular processes. PP2A assembles into diverse trimeric holoenzymes, which consist of a scaffolding (A subunit, a catalytic (C subunit and various regulatory (B subunits. Here we report a 2.0 Å crystal structure of the free B''/PR70 subunit and a SAXS model of an A/PR70 complex. The crystal structure of B''/PR70 reveals a two domain elongated structure with two Ca2+ binding EF-hands. Furthermore, we have characterized the interaction of both binding partner and their calcium dependency using biophysical techniques. Ca2+ biophysical studies with Circular Dichroism showed that the two EF-hands display different affinities to Ca2+. In the absence of the catalytic C-subunit, the scaffolding A-subunit remains highly mobile and flexible even in the presence of the B''/PR70 subunit as judged by SAXS. Isothermal Titration Calorimetry studies and SAXS data support that PR70 and the A-subunit have high affinity to each other. This study provides additional knowledge about the structural basis for the function of B'' containing holoenzymes.

  6. Serum proteins, trace metals and phosphatases in psoriasis

    Bhatnagar M

    1994-01-01

    Full Text Available Serum proteins, zinc, copper, acid phosphatase (AcPase and alkaline phosphatase (AlPase were studied in both active and remission phases of psoriasis. Data were compared with healthy controls, ?1, ? and ? globulins were high in active phase while ?1 and ? globulins were at par in remission phase. Serum copper was low but zinc and alkaline phosphatase were significantly high in both active and remission phases of the disease. Acid phosphatase level was at par in all the experimental groups. Study suggests a positive correlation of globulin, zinc and Alpase in active and remission phase of psoriasis.

  7. Isolation of acetic, propionic and butyric acid-forming bacteria from biogas plants.

    Cibis, Katharina Gabriela; Gneipel, Armin; König, Helmut

    2016-02-20

    In this study, acetic, propionic and butyric acid-forming bacteria were isolated from thermophilic and mesophilic biogas plants (BGP) located in Germany. The fermenters were fed with maize silage and cattle or swine manure. Furthermore, pressurized laboratory fermenters digesting maize silage were sampled. Enrichment cultures for the isolation of acid-forming bacteria were grown in minimal medium supplemented with one of the following carbon sources: Na(+)-dl-lactate, succinate, ethanol, glycerol, glucose or a mixture of amino acids. These substrates could be converted by the isolates to acetic, propionic or butyric acid. In total, 49 isolates were obtained, which belonged to the phyla Firmicutes, Tenericutes or Thermotogae. According to 16S rRNA gene sequences, most isolates were related to Clostridium sporosphaeroides, Defluviitoga tunisiensis and Dendrosporobacter quercicolus. Acetic, propionic or butyric acid were produced in cultures of isolates affiliated to Bacillus thermoamylovorans, Clostridium aminovalericum, Clostridium cochlearium/Clostridium tetani, C. sporosphaeroides, D. quercicolus, Proteiniborus ethanoligenes, Selenomonas bovis and Tepidanaerobacter sp. Isolates related to Thermoanaerobacterium thermosaccharolyticum produced acetic, butyric and lactic acid, and isolates related to D. tunisiensis formed acetic acid. Specific primer sets targeting 16S rRNA gene sequences were designed and used for real-time quantitative PCR (qPCR). The isolates were physiologically characterized and their role in BGP discussed. PMID:26779817

  8. 5, 8, 11, 14-eicosatetraynoic acid suppresses CCL2/MCP-1 expression in IFN-γ-stimulated astrocytes by increasing MAPK phosphatase-1 mRNA stability

    Lee Jee

    2012-02-01

    Full Text Available Abstract Background The peroxisome proliferator-activated receptor (PPAR-α activator, 5,8,11,14-eicosatetraynoic acid (ETYA, is an arachidonic acid analog. It is reported to inhibit up-regulation of pro-inflammatory genes; however, its underlying mechanism of action is largely unknown. In the present study, we focused on the inhibitory action of ETYA on the expression of the chemokine, CCL2/MCP-1, which plays a key role in the initiation and progression of inflammation. Methods To determine the effect of ETYA, primary cultured rat astrocytes and microglia were stimulated with IFN-γ in the presence of ETYA and then, expression of CCL2/MCP-1 and MAPK phosphatase (MKP-1 were determined using RT-PCR and ELISA. MKP-1 mRNA stability was evaluated by treating actinomycin D. The effect of MKP-1 and human antigen R (HuR was analyzed by using specific siRNA transfection system. The localization of HuR was analyzed by immunocytochemistry and subcellular fractionation experiment. Results We found that ETYA suppressed CCL2/MCP-1 transcription and secretion of CCL2/MCP-1 protein through up-regulation of MKP-1mRNA levels, resulting in suppression of c-Jun N-terminal kinase (JNK phosphorylation and activator protein 1 (AP1 activity in IFN-γ-stimulated brain glial cells. Moreover, these effects of ETYA were independent of PPAR-α. Experiments using actinomycin D revealed that the ETYA-induced increase in MKP-1 mRNA levels reflected an increase in transcript stability. Knockdown experiments using small interfering RNA demonstrated that this increase in MKP-1 mRNA stability depended on HuR, an RNA-binding protein known to promote enhanced mRNA stability. Furthermore, ETYA-induced, HuR-mediated mRNA stabilization resulted from HuR-MKP-1 nucleocytoplasmic translocation, which served to protect MKP-1 mRNA from the mRNA degradation machinery. Conclusion ETYA induces MKP-1 through HuR at the post-transcriptional level in a receptor-independent manner. The mechanism

  9. Isolation, characterization and optimization of indigenous acetic acid bacteria and evaluation of their preservation methods

    K Beheshti-Maal

    2010-06-01

    Full Text Available Background and Objectives: Acetic acid bacteria (AAB are useful in industrial production of vinegar. The present study aims at isolation and identification of acetic acid bacteria with characterization, optimization, and evaluation of their acetic acid productivity."nMaterials and Methods: Samples from various fruits were screened for presence of acetic acid bacteria on glucose, yeast extract, calcium carbonate (GYC medium. Carr medium supplemented with bromocresol green was used for distinguishing Acetobacter from Gluconobacter. The isolates were cultured in basal medium to find the highest acetic acid producer. Biochemical tests followed by 16S rRNA and restriction analyses were employed for identification of the isolate and phylogenic tree was constructed. Bacterial growth and acid production conditions were optimized based on optimal inoculum size, pH, temperature, agitation, aeration and medium composition."nResults: Thirty-seven acetic acid bacteria from acetobacter and gluconobacter members were isolated. Acetic acid productivity yielded 4 isolates that produced higher amounts of acid. The highest producer of acid (10.03% was selected for identification. The sequencing and restriction analyses of 16S rRNA revealed a divergent strain of Acetobacter pasteurianus (Gene bank accession number # GU059865. The optimum condition for acid production was a medium composed of 2% glucose, 2% yeast extract, 3% ethanol and 3% acid acetic at inoculum size of 4% at 3L/Min aeration level in the production medium. The isolate was best preserved in GYC medium at 12oC for more than a month. Longer preservation was possible at -70oC."nConclusion: The results are suggestive of isolation of an indigenous acetic acid bacteria. Pilot plan is suggested to study applicability of the isolated strain in acetic acid production.

  10. Isolation and characterisation of lactic acid bacteria from donkey milk.

    Soto Del Rio, Maria de Los Dolores; Andrighetto, Christian; Dalmasso, Alessandra; Lombardi, Angiolella; Civera, Tiziana; Bottero, Maria Teresa

    2016-08-01

    During the last years the interest in donkey milk has increased significantly mainly because of its compelling functional elements. Even if the composition and nutritional properties of donkey milk are known, its microbiota is less studied. This Research Communication aimed to provide a comprehensive characterisation of the lactic acid bacteria in raw donkey milk. RAPD-PCR assay combined with 16S rDNA sequencing analysis were used to describe the microbial diversity of several donkey farms in the North West part of Italy. The more frequently detected species were: Lactobacillus paracasei, Lactococcus lactis and Carnobacterium maltaromaticum. Less abundant genera were Leuconostoc, Enterococcus and Streptococcus. The yeast Kluyveromyces marxianus was also isolated. The bacterial and biotype distribution notably diverged among the farms. Several of the found species, not previously detected in donkey milk, could have an important probiotic activity and biotechnological potential. This study represents an important insight to the ample diversity of the microorganisms present in the highly selective ecosystem of raw donkey milk. PMID:27600975

  11. Antimicrobial susceptibility of lactic acid bacteria isolated from Sombor cheese

    Bulajić Snežana

    2011-01-01

    Full Text Available Extensive literature data pointed out that some lactic acid bacteria (LABs, the predominant microbiota in fermented dairy products, may serve as reservoirs of antibiotic resistance genes potentially transferable to human pathogens. Hence, there is a growing interest in the possible role of Las vectors of antibiotic resistance determinants. This paper reports the susceptibility patterns of a number of Lspecies (belonging to the genera Lactococcus, Lactobacillus, and Enterococcus isolated from different batches of autochthonous Sombor cheese, traditionally made without the addition of starter cultures, and currently proposed as a candidate for PDO/PGI designation. The experimental work was performed to select strains that do not contain antibiotic resistance genes among those with desirable technological characteristics such as rapid acidification, proteolysis, ability to metabolise citrate and form aromogenic compounds. In addition, the results of these screening procedures could also indicate the types and degrees of antimicrobial resistance already present among the Lcommunity of Sombor cheese, which according to their geographically restricted areas of production, specific manufacturing process and characteristic aroma and appearance, represent a distinct ecological niche.

  12. Compartmentation and equilibration of abscisic acid in isolated Xanthium cells

    Bray, E.A.; Zeevaart, J.A.D.

    1986-01-01

    The compartmentation of endogenous abscisic acid (ABA), applied (+/-)-(/sup 3/H)ABA, and (+/-)-trans-ABA was measured in isolated mesophyll cells of the Chicago strain of Xanthium strumarium L. The release of ABA to the medium in the presence or absence of DMSO was used to determine the equilibration of ABA in the cells. It was found that a greater percentage of the (+/-)-(/sup 3/H)ABA and the (+/-)-trans-ABA was released into the medium than of the endogenous ABA, indicating that applied ABA did not equilibrate with the endogenous material. Therefore, in further investigations only the compartmentation of endogenous ABA was studied. Endogenous ABA was released from Xanthium cells according to the pH gradients among the various cellular compartments. Thus, darkness, high external pH, KNO/sub 2/, and drought-stress all increased the efflux of ABA from the cells. Efflux of ABA from the cells in the presence of 0.6 M mannitol occurred within 30 seconds, but only 8% of the endogenous material was released during the 20 minute treatment.

  13. Taxonomic homogeneity of a salt-tolerant lactic acid bacteria isolated from shoyu mash.

    Hanagata, Hiroshi; Shida, Osamu; Takagi, Hiroaki

    2003-04-01

    Forty-seven salt-tolerant lactic acid bacteria, which had been isolated from different places and grown in 15% NaCl, were examined to assess their taxonomic heterogeneity. Among the isolates, 42 were isolated from shoyu mash during the acid fermentation phase, 2 were from miso and 3 were from anchovy pickles. All isolates were identified as Tetragenococcus halophilus on the basis of DNA relatedness values. We further examined 102 phenotypic characteristics of them. The isolates exhibited differences in only 16, supporting the conclusion obtained from the DNA relatedness analysis. PMID:12833212

  14. Antimicrobial properties of lactic acid bacteria isolated from uruguayan artisan cheese

    Martín Fraga Cotelo; Karen Perelmuter Schein; Sheila Solange Giacaman Salvo; Pablo Miguel Zunino Abirad; Silvana Beatriz Carro Techera

    2013-01-01

    Uruguayan artisan cheese is elaborated with raw milk and non-commercial starters. The associated native microbiota may include lactic acid bacteria and also potentially pathogenic bacteria. Lactic acid bacteria were isolated from artisan cheese, raw milk, and non-commercial starter cultures, and their potential bacteriocin production was assessed. A culture collection of 509 isolates was obtained, and five isolates were bacteriocin-producers and were identified as Enterococcus durans,Lactobac...

  15. Isolation and cellular fatty acid composition of psychrotrophic Halomonas strains from Antarctic sea water

    Vipra Vijay Jadhav; Amit Yadav; Shouche, Yogesh S.; Rama Kaustubh Bhadekar

    2013-01-01

    Microorganisms from extreme environments such as Arctic, Antarctic and Polar regions modulate their membrane fatty acids to survive in such habitats. Characterization of such microorganisms helps in understanding their physiological behavior. In view of this, the present article describes isolation, characterization and cellular fatty acid composition of three bacterial isolates from Antarctic sea water samples. All the three isolates (BRI 6, 29 and 31) were psychrotrophic Gram negative rods....

  16. Assessment and kinetics of soil phosphatase in Brazilian Savanna systems.

    Ferreira, Adão S; Espíndola, Suéllen P; Campos, Maria Rita C

    2016-05-31

    The activity and kinetics of soil phosphatases are important indicators to evaluate soil quality in specific sites such as the Cerrado (Brazilian Savanna). This study aimed to determine the activity and kinetic parameters of soil phosphatase in Cerrado systems. Soil phosphatase activity was assessed in samples of native Cerrado (NC), no-tillage (NT), conventional tillage (CT) and pasture with Brachiaria brizantha (PBb) and evaluated with acetate buffer (AB), tris-HCl buffer (TB), modified universal buffer (MUB) and low MUB. The Michaelis-Menten equation and Eadie-Hofstee model were applied to obtain the kinetic parameters of soil phosphatase using different concentrations of p-nitrophenol phosphate (p-NPP). MUB showed the lowest soil phosphatase activity in all soils whereas AB in NC and NT presented the highest. Low MUB decreased interferences in the assessment of soil phosphatase activity when compared to MUB, suggesting that organic acids interfere on the soil phosphatase activity. In NC and NT, soil phosphatase activity performed with TB was similar to AB and low MUB. Km values from the Michaels-Menten equation were higher in NC than in NT, which indicate a lower affinity of phosphatase activity for the substrate in NC. Vmax values were also higher in NC than in NT. The Eadie-Hofstee model suggests that NC had more phosphatase isoforms than NT. The study showed that buffer type is of fundamental importance when assessing soil phosphatase activity in Cerrado soils. PMID:27254453

  17. Isolation of lactic acid bacteria with potential protective culture characteristics from fruits

    Hashim, Nurul Huda; Sani, Norrakiah Abdullah

    2015-09-01

    Lactic acid bacteria are also known as beneficial microorganisms abundantly found in fermented food products. In this study, lactic acid bacteria were isolated from fresh cut fruits obtained from local markets. Throughout the isolation process from 11 samples of fruits, 225 presumptive lactic acid bacteria were isolated on MRS agar medium. After catalase and oxidase tests, 149 resulted to fit the characteristics of lactic acid bacteria. Further identification using Gram staining was conducted to identify the Gram positive bacteria. After this confirmation, the fermentation characteristics of these isolates were identified. It was found that 87 (58.4%) isolates were heterofermentative, while the rest of 62 (41.6%) are homofermentative lactic acid bacteria. Later, all these isolates were investigated for the ability to inhibit growth of Staphylococcus aureus using agar spot assay method. Seven (4.7%) isolates showed strong antagonistic capacity, while 127 (85.2%) and 8 (5.4%) isolates have medium and weak antagonistic capacity, respectively. The other 7 (4.7%) isolates indicated to have no antagonistic effect on S. aureus. Results support the potential of LAB isolated in this study which showed strong antagonistic activity against S. aureus may be manipulated to become protective cultures in food products. While the homofermentative or heterofermentative LAB can be utilized in fermentation of food and non-food products depending on the by-products required during the fermentation.

  18. Comparative analysis of antimicrobial and proteolytic activity of lactic acid bacteria isolated from Zlatar cheese

    Topisirović Ljubiša; Veljović Katarina; Terzić-Vidojević Amarela; Strahinić Ivana; Kojić Milan

    2007-01-01

    Traditional artisan Zlatar cheese belongs to the group of white, semi hard home-made cheeses, which are produced from no pasteurized cow's milk, without addition of any known bacterial starter culture. In total, 253 Gram-positive and catalase negative lactic acid bacteria (LAB) were isolated. Results showed that 70 out of 253 analyzed isolates produced antimicrobial compounds known as bacteriocins. Most isolates from genera Lactococcus and Enterococcus, and isolates belonging to ...

  19. Relationship of spermatoscopy, prostatic acid phosphatase activity and prostate-specific antigen (p30) assays with further DNA typing in forensic samples from rape cases.

    Romero-Montoya, Lydia; Martínez-Rodríguez, Hugo; Pérez, Miguel Antonio; Argüello-García, Raúl

    2011-03-20

    In the forensic laboratory the biological analyses for rape investigation commonly include vaginal swabs as sample material combined to biochemical tests including sperm cytology (SC) and detection of acid phosphatase activity (AP) and prostate-specific antigen (PSA, p30) for the conclusive identification of semen components. Most reports comparing these tests relied on analysis of semen samples or donor swabs taken under controlled conditions; however their individual or combined efficacy under real live sampling conditions in different laboratories is largely unknown. We carried out SC, APA and PSA analyses in vaginal swabs collected from casework rapes submitted to Mexican Forensic Laboratories at Texcoco and Toluca. On the basis of positive and negative results from each assay and sample, data were classified into eight categories (I-VIII) and compared with those obtained in the two only similar studies reported in Toronto, Canada and Hong Kong, China. SC and APA assays had the higher overall positivity in Toluca and Texcoco samples respectively and otherwise PSA had a lower but very similar positivity between these two laboratories. When compared to the previous studies some similarities were found, namely similar frequencies (at a ratio of approximately 1 out of 3) of samples being positive or negative by all techniques (Categories I and VI respectively) and a comparable overall positivity of APA and SC but higher than that of PSA. Indeed the combined results of using SC, APA and PSA tests was considered as conclusive for semen detection from approximately 1 out of 3 cases (Category I) to approximately 1 out of 2 cases in a scenario where at least SC is positive, strongly presumptive in 2 out of 3 cases (with at least one test positive) and the remainder 1 out of 3 cases (Category VI) suggested absence of semen. By determining Y-STR polymorphisms (12-loci) in additional samples obtained at Toluca laboratory, complete DNA profiles were determined from all

  20. Age-related changes of serum tartrate-resistant acid phosphatase 5b and the relationship with bone mineral density in Chinese women

    Yue-juan QIN; Zhen-lin ZHANG; Hao ZHANG; Wei-wei HU; Yu-juan LIU; Yun-qiu HU; Miao LI; Jie-mei GU; Jin-wei HE

    2008-01-01

    Aim: Ostcoclastic activity is mainly assessed by measurement of urinary markers (eg C-terminal cross-linked telopeptides of type I collagen, N-terminal cross-linked telopeptides of type I collagen etc), the levels of which could often be affected by renal clearance. Recently, serum tartrate-resistant acid phosphatase 5b (TRACP5b) has been used as an alternative serum marker to evaluate osteoclastic activity. We investigated the age-related changes of TRACP5b level and its association with bone mineral density (BMD) in Chinese women. Methods: Seven-hundred and twenty-two Chinese mainland women aged 20-79 years were recruited in the study. Serum TRACP5b level was measured using immunoassay to evaluate the state of bone resorption. Bone mineral density (BMD) (g/cm2) at lumbar spine 1-4 and proximal femur were measured by duel-energy X-ray absorptiometry. Results: The serum TRACP5b level reached a bottom value in premenopausal women aged 30-39, gradually increased in women aged 40-49, rapidly rose in women aged 50-59, and culminated with a maximum value in women aged 60-69 before a slow drop in women aged 70-79. The average level of TRACPSb was significantly higher in postmenopausal women [(3.29±1.07) U/L] than in premenopausal women ([1.70±0.59] U/L). The levels of TRACP5b were inversely correlated with BMD at all measured sites (P<0.001). Furthermore, the level of TRACP5b was obviously higher in women with osteoporosis and osteopenia than those with normal bone mass (P<0.001). Conclusion: We have established the reference values of serum TRACPSb in Chinese mainland women, and found that postmenopausal women had higher TRACP5b concentration than younger women. The results showed that serum TRACPSb was a sensitive and useful parameter for the evaluation of age-related changes of bone absorption.

  1. Change of glutamic acid decarboxylase antibody and protein tyrosine phosphatase antibody in Chinese patients with acute-onset type 1 diabetes mellitus

    CHAO Chen; HUANG Gan; LI Xia; YANG Lin; LIN Jian; JIN Ping; LUO Shuo-ming

    2013-01-01

    Background Glutamic acid decarboxylase antibody (GADA) and protein tyrosine phosphatase antibody (IA-2A) are two major autoantibodies,which exert important roles in the process of type 1 diabetes mellitus (T1D).Our study aimed to investigate the changes in positivity and titers of GADA and IA-2A during the course of Chinese acute-onset T1D patients and their relationships with clinical features.Methods Two hundreds and forty-seven Chinese newly diagnosed acute-onset T1D patients were consecutively recruited.GADA and IA-2A were detected at the time of diagnosis,one year later,3-5 years later after diagnosis during the follow-up; all the clinical data were recorded and analyzed as well.Results During the course of acute-onset T1D,the majority of patients remained stable for GADA or IA-2A,however,a few patients changed from positivity to negativity and fewer patients converted from negativity to positivity.The prevalence of GADA was 56.3% at diagnosis,decreasing to 50.5% one year later,and 43.3% 3-5 years later while the corresponding prevalence of IA-2A were 32.8%,31.0% and 23.3%,respectively.The median GADA titers were 0.0825 at diagnosis,declining to 0.0585 one year later and 0.0383 3-5 years later (P <0.001),while the corresponding median titers were 0.0016,0.0010,0.0014 for IA-2A,respectively.Fasting C-peptide (FCP) and postprandial C-peptide 2 hours (PCP2h)levels of GADA or IA-2A negativity persistence patients were higher than those of positivity persistence and negativity conversion patients (P <0.05) which indicated GADA or IA-2A negativity persistence T1D patients had a less loss of β cell function.Conclusion Our data suggest that repeated detection of GADA and IA-2A are necessary for differential diagnosis of autoimmune diabetes and the indirect prediction of the β cell function in Chinese patients.

  2. Persistently increased intestinal fraction of alkaline phosphatase

    Nathan, E; Baatrup, G; Berg, H;

    1984-01-01

    Persistent elevation of the intestinal fraction of the alkaline phosphatase (API) as an isolated finding has to our knowledge not been reported previously. It was found in a boy followed during a period of 5.5 years. The only symptom was transient periodic fatigue observed at home, but not apparent...

  3. Isolation, structure, and synthesis of viridic acid, a new tetrapeptide mycotoxin of Penicillium viridicatum Westling

    The isolation of a new toxic metabolite, viridic acid, from Penicillium viridicatum Westling is described. The chemical and spectroscopic properties of the compound are interpreted in terms of the tetrapeptide structure (N,N-dimethyl-o-aminobenzoyl)-glycyl-(N'-methyl-L-valyl)-o-aminobenzoic acid. The structure and chirality of viridic acid were confirmed by total synthesis

  4. Concentrations of testosterone, luteal hormone and prolactin in the serum as well as comparisons of sensitivity between radioimmunoassays and enzyme assays for the detection of acid prostate phosphatase in the presence of carcinomas of the prostate

    The relationship between carcinomas of the prostate and the plasma levels of testosterone, luteal hormone and prolactin as well as the possible influence of these neoplasms on the testosterone binding capacity and free testosterone index are investigated for various tumour stages and degrees of histological differentiation, in connection with several forms of local therapy as well as a variety of contrasexual methods. The sensitivity of enzyme assays and radioimmunoassays for the detection of acid prostate phosphatase is evaluated within the framework of this study. (MBL)

  5. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either 3H-fatty acids or [3H]ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the 3H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of [3H]ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from 3H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the 3H-fatty acid and the [3H]ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the [3H]ethanolamine label from the purified alkaline phosphatase. The 3H radioactivity in alkaline phosphatase purified from [3H]ethanolamine-labeled cells comigrated with authentic [3H]ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the 3H-fatty acid and [3H]ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase

  6. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Jemmerson, R.; Low, M.G.

    1987-09-08

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  7. Rhizobiales-like Phosphatase 2 from Arabidopsis thaliana Is a Novel Phospho-tyrosine-specific Phospho-protein Phosphatase (PPP) Family Protein Phosphatase.

    Uhrig, R Glen; Labandera, Anne-Marie; Muhammad, Jamshed; Samuel, Marcus; Moorhead, Greg B

    2016-03-11

    Cellular signaling through protein tyrosine phosphorylation is well established in mammalian cells. Although lacking the classic tyrosine kinases present in humans, plants have a tyrosine phospho-proteome that rivals human cells. Here we report a novel plant tyrosine phosphatase from Arabidopsis thaliana (AtRLPH2) that, surprisingly, has the sequence hallmarks of a phospho-serine/threonine phosphatase belonging to the PPP family. Rhizobiales/Rhodobacterales/Rhodospirillaceae-like phosphatases (RLPHs) are conserved in plants and several other eukaryotes, but not in animals. We demonstrate that AtRLPH2 is localized to the plant cell cytosol, is resistant to the classic serine/threonine phosphatase inhibitors okadaic acid and microcystin, but is inhibited by the tyrosine phosphatase inhibitor orthovanadate and is particularly sensitive to inhibition by the adenylates, ATP and ADP. AtRLPH2 displays remarkable selectivity toward tyrosine-phosphorylated peptides versus serine/threonine phospho-peptides and readily dephosphorylates a classic tyrosine phosphatase protein substrate, suggesting that in vivo it is a tyrosine phosphatase. To date, only one other tyrosine phosphatase is known in plants; thus AtRLPH2 represents one of the missing pieces in the plant tyrosine phosphatase repertoire and supports the concept of protein tyrosine phosphorylation as a key regulatory event in plants. PMID:26742850

  8. Radiolytic degradation of sorbic acid in isolated systems

    Effect of Co(60) gamma-irradiation on stability of sorbic acid (SA) in solutions, dough and chapaties has been investigated. SA was highly susceptible to radiolytic degradation in aqueous systems. Rate of degradation decreased with rise in pH. Sugars, hydrocolloids except pectin, citric acid, lactic acid, malic acid, arginine and threonine, catalyzed degradation while oxalic acid, maleic acid, Cu2+, nitrite, nitrate and phthalate had protective effects. SA was more stable in alcohols and vegetable oils than in aqueous solutions. In wheat flour radiolytic degradation of SA was less at lower moisture. Relatively SA was more stable in chapaties than in dough. Gelatinization and addition of oil in dough reduced degradation of SA

  9. Colistin-oxolinic acid blood agar: a selective medium for the isolation of Gardnerella vaginalis.

    Thompson, J. S.

    1985-01-01

    Colistin-oxolinic acid medium is proposed as a selective isolation medium for Gardnerella vaginalis. The medium is effective in inhibiting staphylococci and gram-negative bacteria while allowing growth of G. vaginalis.

  10. 基于不同方法测定土壤酸性磷酸酶活性的比较%Comparison of soil acid phosphatase activity determined by different methods

    李莹飞; 耿玉清; 周红娟; 杨英

    2016-01-01

    土壤酸性磷酸酶与有机磷的矿化及植物的磷素营养关系最为密切。目前国内学者在测定酸性磷酸酶活性时主要参照关松荫《土壤酶及其研究法》中以磷酸苯二钠为基质的测定方法,而国外学者主要参照 Dick《Methods of Soil Enzymology》中以对硝基苯磷酸二钠为基质的测定方法(PNPP)。但是,在以磷酸苯二钠为基质测定生成物的过程中,常出现显色程度不明显的问题;另外,采用不同基质测定酸性磷酸酶活性也造成了测定方法选择的困难。为合理选择土壤酸性磷酸酶活性的测定方法,本研究选用酸性、中性和碱性土壤各10个土样,分别采用以磷酸苯二钠为基质,且在显色阶段分别加入 pH5.0醋酸盐缓冲液(DPP 1)和 pH9.4硼酸盐缓冲液(DPP 2)的方法,以及PNPP方法测定土壤酸性磷酸酶活性。同时也研究了不同pH缓冲液和苯酚浓度对生成物显色反应的影响。结果表明:以磷酸苯二钠为基质、在显色反应阶段加入 pH≤6的缓冲液时,苯酚和2,6-二溴苯醌氯亚胺不显色;当加入pH≥8的缓冲液时,两者之间显色且苯酚浓度和吸光值的Pearson相关系数极显著。这说明 pH 低是导致高苯酚浓度和2,6-二溴苯醌氯亚胺显色效果差的一个主要原因。此外,采用PNPP 方法测定时,在酸性、中性和碱性土壤中,10个样本酸性磷酸酶活性的变异系数分别较 DPP 2增加了70.04%、42.44%和21.17%;极差分别是DPP 2的27.18倍、26.85倍和39.43倍。总之,如果选用磷酸苯二钠为基质测定土壤酸性磷酸酶活性,应在显色阶段加入碱性硼酸盐缓冲液;选用对硝基苯磷酸二钠为基质,是更为简单和灵敏的方法。%Soil phosphatase, especially acid phosphatase, plays a critical role in the decomposition of organic phosphorus and has a major impact on plant phosphorus uptake. Most Chinese researchers refer to the book entitled Soil Enzyme and Its Research

  11. The isolation and improvement of Aspergillus niger by radiation for higher production of citric acid

    Local citric acid producer of fungal strain Aspergillus niger have been successfully isolated from stale bread and onion. The isolates, designated as SB 1 and NN I showed a potential performance for citric acid production of 49% and 52% yield respectively, in shake flask studies. The strain improvement on NN1 was carried out by radiation induced mutation by gamma rays at LD50 of 1.28 kGy

  12. Naturally fermented Jijelian black olives: microbiological characteristics and isolation of lactic acid bacteria

    Karam, Nour-Eddine; Leghouchi, Essaid; Boudjerda, Jamel; Idoui, Tayeb

    2009-01-01

    A study of the microflora of traditionally fermented black olives in Eastern Algeria is presented. A count of the following microbial groups was carried out: mesophilic bacteria, enterobacteria, lactic acid bacteria (LAB), staphylococci and yeast. In a second phase, the identification and assessment of the technological traits of LAB was performed. Seventeen lactic acid bacteria were isolated and identified. These isolates were represented by two genera: Lactobacillus and Leuconostoc. The res...

  13. ISOLATION OF LACTIC ACID BACTERIA UNDER LOW TEMPERATURE FOR THE PREPARATION OF YOGURT

    Javid Ahmad Bhat; Mohd Irfan Naik; R.K Tenguria

    2014-01-01

    An investigation of isolation of Lactic acid bacteria was carried out under low temperature for the preparation of Yogurt by using various food supplements like carrot, ground-nut and tomato juices. Methods: Various samples of Cow milk, Skimmed milk were processed along with nutrients like Carrot, ground nut and tomato juices with Tryptone glucose yeast extract agar (TGYA) at different temperatures like 50C, 150C and 220C for the isolation of Lactic acid bacteria for the preparation of yogurt...

  14. Partial purification and characterization of phosphotyrosyl-protein phosphatase(s) from human erythrocyte cytosol

    Phosphotyrosyl-protein phosphatase activity of human erythrocyte cytosol can be resolved into two fractions by DEAE-cellulose chromatography followed by P-cellulose chromatography. Both 32P-Tyr-phosphatases are able to dephosphorylate 32P-Tyr of poly (Glu-Tyr) 4:1 but no angiotensin II and synthetic peptide Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Arg-Gly, previously phosphorylated on tyrosine residues by rat spleen tyrosine-protein kinase. Both 32P-Tyr-phosphatase activities distinctly differ from either 32P-Ser-casein phosphatase activity or acid and alkaline p-nitrophenylphosphatase activities with regard to catalytic and physico-chemical properties such as substrate specificity, chromatographic behavior, response to various effectors

  15. Isolation and identification of phosphatidic acid targets from plants.

    C. Testerink; H.L. Dekker; Z.-Y. Lim; M.K. Johns; A.B. Holmes; C.G. de Koster; N.T. Ktisakis; T. Munnik

    2004-01-01

    Phosphatidic acid (PA) is emerging as an important lipid signalling molecule. In plants, it is implicated in various stress-signalling pathways and is formed in response to wounding, osmotic stress, cold stress, pathogen elicitors, Nod factors, ethylene and abscisic acid. How PA exerts its effects i

  16. Lactic acid bacteria against post-harvest moulds and ochratoxin A isolated from stored wheat

    Belkacem-Hanfi, N.; Fhoula, I.; Semmar, N.; Guesmi, A.; Perraud Gaime, Isabelle; Ouzari, H. I.; A. Boudabous; Roussos, Sevastianos

    2014-01-01

    A total of 54 lactic acid bacteria (LAB) were isolated from stored wheat samples sourced from grain silos in North Tunisia. Fifteen representative isolates were identified by 16S rDNA sequencing as Pediococcus pentosaceus, Lactobacillus plantarum, Lactobacillus graminis, Lactobacillus coryniformis and Weissella cibaria. These isolates were screened for antifungal activity in dual culture agar plate assay against eight post-harvest moulds (Penicillium expansum, Penicillium chrysogenum, Penicil...

  17. Production of bacteriocin-like substances by lactic acid bacteria isolated from regional ovine cheese

    Cássia Regina Nespolo; Adriano Brandelli

    2010-01-01

    Lactic acid bacteria (LAB) were isolated from ovine milk and cheeses manufactured in the South Region of Brazil. Among 112 bacterial isolates investigated, 59 were chosen through a screening for LAB. Among these 59 strains of LAB, 21% showed antimicrobial, proteolytic and lipolytic activities. Based on this screening, Lactobacillus plantarum LCN 17 and Lactobacillus rhamnosus LCN 43 were selected and tested for the production of bacteriocin-like substances (BLS). The BLS produced by both isol...

  18. Comparative genetic analysis of Arabidopsis purple acid phosphatases AtPAP10, AtPAP12, and AtPAP26 provides new insights into their roles in plant adaptation to phosphate deprivation

    Liangsheng Wang; Shan Lu; Ye Zhang; Zheng Li; Xiaoqiu Du; Dong Liu

    2014-01-01

    Induction and secretion of acid phosphatases (APases) is thought to be an adaptive mechanism that helps plants survive and grow under phosphate (Pi) deprivation. In Arabidopsis, there are 29 purple acid phosphatase (AtPAP) genes. To systematical y investigate the roles of different AtPAPs, we first identified knockout or knock-down T-DNA lines for al 29 AtPAP genes. Using these atpap mutants combined with in-gel and quantitative APase enzyme assays, we demonstrated that AtPAP12 and AtPAP26 are two major intracellular and secreted APases in Arabidopsis while AtPAP10 is mainly a secreted APase. On Pi-deficient (P-) medium or P-medium supplemented with the organophosphates ADP and fructose-6-phosphate (Fru-6-P), growth of atpap10 was significantly reduced whereas growth of atpap12 was only moderately reduced, and growth of atpap26 was nearly equal to that of the wild type (WT). Overexpression of the AtPAP12 or AtPAP26 gene, however, caused plants to grow better on P-or P- medium supplemented with ADP or Fru-6-P. Interest-ingly, Pi levels are essential y the same for the WT and overexpressing lines, although these two types of plants have significantly different growth phenotypes. These results suggest that the APases may have other roles besides enhancing internal Pi recycling or releasing Pi from external organophosphates for plant uptake.

  19. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    The overall objective of this project is to examine the activity of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium U(VI) phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO43- as a means to detoxify radionuclides and heavy metals. An experimental approach was designed to determine the extent of phosphatase activity in bacteria previously isolated from contaminated subsurface soils collected at the ERSP Field Research Center (FRC) in Oak Ridge, TN. Screening of 135 metal resistant isolates for phosphatase activity indicated the majority (75 of 135) exhibited a phosphatase-positive phenotype. During this phase of the project, a PCR based approach has also been designed to assay FRC isolates for the presence of one or more classes of the characterized non-specific acid phophastase (NSAP) genes likely to be involved in promoting U(VI) precipitation. Testing of a subset of Pb resistant (Pbr) Arthrobacter, Bacillus and Rahnella strains indicated 4 of the 9 Pbr isolates exhibited phosphatase phenotypes suggestive of the ability to bioprecipitate U(VI). Two FRC strains, a Rahnella sp. strain Y9602 and a Bacillus sp. strain Y9-2, were further characterized. The Rahnella sp. exhibited enhanced phosphatase activity relative to the Bacillus sp. Whole-cell enzyme assays identified a pH optimum of 5.5, and inorganic phosphate accumulated in pH 5.5 synthetic groundwater (designed to mimic FRC conditions) incubations of both strains in the presence of a model organophosphorus substrate provided as the sole C and P source. Kinetic experiments showed that these two organisms can grow in the presence of 200 (micro)M dissolved uranium and that Rahnella is much more efficient in precipitating U(VI) than Bacillus sp. The

  20. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    Robert J. Martinez; Melanie J. Beazley; Samuel M. Webb; Martial Taillefert (co-PI); and Patricia A. Sobecky

    2007-04-19

    The overall objective of this project is to examine the activity of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium [U(VI)] phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO4 3- as a means to detoxify radionuclides and heavy metals. An experimental approach was designed to determine the extent of phosphatase activity in bacteria previously isolated from contaminated subsurface soils collected at the ERSP Field Research Center (FRC) in Oak Ridge, TN. Screening of 135 metal resistant isolates for phosphatase activity indicated the majority (75 of 135) exhibited a phosphatase-positive phenotype. During this phase of the project, a PCR based approach has also been designed to assay FRC isolates for the presence of one or more classes of the characterized non-specific acid phophastase (NSAP) genes likely to be involved in promoting U(VI) precipitation. Testing of a subset of Pb resistant (Pbr) Arthrobacter, Bacillus and Rahnella strains indicated 4 of the 9 Pbr isolates exhibited phosphatase phenotypes suggestive of the ability to bioprecipitate U(VI). Two FRC strains, a Rahnella sp. strain Y9602 and a Bacillus sp. strain Y9-2, were further characterized. The Rahnella sp. exhibited enhanced phosphatase activity relative to the Bacillus sp. Whole-cell enzyme assays identified a pH optimum of 5.5, and inorganic phosphate accumulated in pH 5.5 synthetic groundwater (designed to mimic FRC conditions) incubations of both strains in the presence of a model organophosphorus substrate provided as the sole C and P source. Kinetic experiments showed that these two organisms can grow in the presence of 200 μM dissolved uranium and that Rahnella is much more efficient in precipitating U(VI) than Bacillus sp. The

  1. Improved method for simultaneous isolation of proteins and nucleic acids.

    Chey, Soroth; Claus, Claudia; Liebert, Uwe Gerd

    2011-04-01

    Guanidinium thiocyanate-phenol-chloroform extraction (GTPC extraction) is widely used in molecular biology for isolating DNA, RNA, and proteins. Protein isolation by commercially available GTPC solutions is time consuming and the resulting pellets are only incompletely soluble. In this study ethanol-bromochloropropane-water was used for precipitation of proteins from the phenol-ethanol phase after GTPC extraction of RNA and DNA. The precipitated proteins can be readily dissolved in 4% SDS for subsequent analysis. This technique allows a fast (30min) and efficient (with a protein recovery of up to 95%) extraction of proteins for the study of transcriptional and posttranscriptional events from the same sample. PMID:21094121

  2. Effect of phenylmercuric acetate injections on phosphatase activity in chickens resistant and susceptible to Leukosis

    Miller, V.L.; Bearse, G.E.; Csonka, E.

    1972-01-01

    The weighted means of liver and kidney alkaline phosphatase activity was greater in three strains of chickens classified as susceptible to limphoid leukosis than in five strains classified as resistant. On the same basis, four strains classified as susceptible to Marek's disease had more liver alkaline phosphatase activity than four strains classified as resistant. The weighted means of liver and kidney acid phosphatase activity were not different among the same strains of chickens classified similarly. Kidney alkaline phosphatase activity was the most generally inhibited by phenylmercuric acetate injections, followed by liver acid and alkaline phosphatase. Kidney acid phosphatase activity was enhanced by phenylmercuric acetate injections in three strains of chickens classified as resistant to both lymphoid leukosis and Marek's disease. Liver acid phosphatase activity was depressed in three strains classed as resistant to lymphoid leukosis.

  3. Antimicrobial properties of lactic acid bacteria isolated from uruguayan artisan cheese

    Martín Fraga Cotelo

    2013-12-01

    Full Text Available Uruguayan artisan cheese is elaborated with raw milk and non-commercial starters. The associated native microbiota may include lactic acid bacteria and also potentially pathogenic bacteria. Lactic acid bacteria were isolated from artisan cheese, raw milk, and non-commercial starter cultures, and their potential bacteriocin production was assessed. A culture collection of 509 isolates was obtained, and five isolates were bacteriocin-producers and were identified as Enterococcus durans,Lactobacillus casei, and Lactococcus lactis. No evidence of potential virulence factors were found in E. durans strains. These are promising results in terms of using these native strains for cheese manufacture and to obtain safe products.

  4. Comparative analysis of antimicrobial and proteolytic activity of lactic acid bacteria isolated from Zlatar cheese

    Topisirović Ljubiša

    2007-01-01

    Full Text Available Traditional artisan Zlatar cheese belongs to the group of white, semi hard home-made cheeses, which are produced from no pasteurized cow's milk, without addition of any known bacterial starter culture. In total, 253 Gram-positive and catalase negative lactic acid bacteria (LAB were isolated. Results showed that 70 out of 253 analyzed isolates produced antimicrobial compounds known as bacteriocins. Most isolates from genera Lactococcus and Enterococcus, and isolates belonging to species Lactobacillus plantarum and Lb. brevis, do not synthesize extracellular proteinase. In contrast, isolates from subspecies Lb. paracasei subsp. paracasei showed very good proteolytic activity. It was observed that good proteolytic activity of isolates was not in correlation with their good antimicrobial activity in the most of isolates.

  5. Detection of plasmid deoxyribonucleic acid in an isolate of Lactobacillus acidophilus.

    Klaenhammer, T R; Sutherland, S M

    1980-01-01

    Eight strains of Lactobacillus acidophilus were examined for the presence of plasmid deoxyribonucleic acid, and one, a pig intestinal isolate, showed the presence of a 13.7- and a 6.3-megadalton plasmid. This is the first reported evidence for plasmid deoxyribonucleic acid in Lactobacillus acidophilus. The functions of these plasmids are presently unknown.

  6. Absence of acetohydroxy acid synthetase in a clinical isolate of Neisseria gonorrhoeae requiring isoleucine and valine.

    Lerner, S A; Friedman, E L; Dudek, E J; Kominski, G; Bohnhoff, M.; Morello, J A

    1980-01-01

    A clinical isolate of Neisseria gonorrhoeae with an unusual growth requirement for isoleucine and valine lacked the activity of acetohydroxy acid synthetase, one of the enzymes required for the biosynthesis of these amino acids. A spontaneous mutant which no longer required isoleucine and valine had acquired this enzymatic activity.

  7. The extended human PTPome: a growing tyrosine phosphatase family.

    Alonso, Andrés; Pulido, Rafael

    2016-04-01

    Tyr phosphatases are, by definition, enzymes that dephosphorylate phospho-Tyr (pTyr) from proteins. This activity is found in several structurally diverse protein families, including the protein Tyr phosphatase (PTP), arsenate reductase, rhodanese, haloacid dehalogenase (HAD) and His phosphatase (HP) families. Most of these families include members with substrate specificity for non-pTyr substrates, such as phospho-Ser/phospho-Thr, phosphoinositides, phosphorylated carbohydrates, mRNAs, or inorganic moieties. A Cys is essential for catalysis in PTPs, rhodanese and arsenate reductase enzymes, whereas this work is performed by an Asp in HAD phosphatases and by a His in HPs, via a catalytic mechanism shared by all of the different families. The category that contains most Tyr phosphatases is the PTP family, which, although it received its name from this activity, includes Ser, Thr, inositide, carbohydrate and RNA phosphatases, as well as some inactive pseudophosphatase proteins. Here, we propose an extended collection of human Tyr phosphatases, which we call the extended human PTPome. The addition of new members (SACs, paladin, INPP4s, TMEM55s, SSU72, and acid phosphatases) to the currently categorized PTP group of enzymes means that the extended human PTPome contains up to 125 proteins, of which ~ 40 are selective for pTyr. We set criteria to ascribe proteins to the extended PTPome, and summarize the more important features of the new PTPome members in the context of their phosphatase activity and their relationship with human disease. PMID:26573778

  8. Production of conjugated linoleic acids by Lactobacillus plantarum strains isolated from naturally fermented Chinese pickles*

    Liu, Pei; Shen, Sheng-rong; Ruan, Hui; ZHOU, Qian; Ma, Liu-liu; He, Guo-qing

    2011-01-01

    Naturally fermented pickles harbour many lactic acid bacteria (LAB). Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, lp15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 µg/ml of linoleic acid (LA). Compa...

  9. Isolation and identification of indigenous lactic acid bacteria from North Sumatra river buffalo milk

    Heni Rizqiati

    2015-06-01

    Full Text Available Buffalo milk is a source of various lactic acid bacteria (LAB which is potential as culture starter as well as the probiotic. This study was conducted to isolate and identify LAB from indigenous North Sumatra river buffalo milk. Lactic acid bacteria was isolated and grown in medium De Man Rogosa Sharpe Agar (MRSA. The isolation was conducted to obtain pure isolate. The identification of LAB was studied in terms of morphology, physiology, biochemistry and survival on low pH. Morphology tests were conducted by Gram staining and cell forming; physiology tests were conducted for growing viability at pH 4.5 and temperature at 45oC; whereas biochemistry tests were conducted for CO2, dextran and NH3 productions. Determination of LAB species was conducted using Analytical Profile Index (API test CHL 50. Results of identification showed that 41 isolates were identified as LAB with Gram-positive, catalase-negative, rod and round shaped characteristics. Resistance test done to low pH (pH 2 for the lactic acid bacteria showed decrease of bacteria viability up to1.24±0.68 log cfu/ml. The resistant isolates at low pH were L12, L16, L17, L19, L20, M10, P8, S3, S19 and S20. Identification with API test CHL 50 for 10 isolates showed that four isolates were identified as Lactobacillus plantarum, L. brevis, L. pentosus and Lactococuslactis.

  10. Effects of indole-3-acetic acid on Botrytis cinerea isolates obtained from potted plants.

    Martínez, J A; Valdés, R; Gómez-Bellot, M J; Bañón, S

    2011-01-01

    We study the growth of different isolates of Botrytis cinerea collected from potted plants which were affected by Botrytis blight in southern Spain during recent years. These isolates, which show widely phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid applications and paclobutrazol, an efficient plant growth retardant and fungicide at the same time. In this work, we have evaluated the effect of the auxin indole-3-acetic acid (IAA) dose (0, 1, 10, and 100 mg/plate) on the growth of the collection of B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantona camara, and Lonicera japonica. B. cinerea produces indolacetic acid, but so far the precise biosynthetic pathway and some effects on this fungal species are still unclear, although recent studies have revealed an antifungal activity of IAA on several fungi, including B. cinerea isolated from harvested fruits. Mycelial growth curves and growth rates assessed from difference in colony areas during the both linear and deceleration phase, conidiation (measured as time of appearance), conidia length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 35 days. Mycelial growth curves fitted a typical kinetic equation of fungi grown on solid media. B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and auxin dose. This plant growth substance delayed mycelial growth during the linear phase in an isolate-dependent manner, thus isolates from C. persicum, H. macrophylla and L. camara were more affected by IAA than L. japonica. On the other hand, 100 mg of IAA was the critical dose to significantly reduce the growth rate in all isolates and to promote brown-striped hyphae development, especially in isolate from C. persicum. 10 and 100 mg

  11. Optimization of petroleum acid isolation from lower oil fractions of Vojvodina "Velebit" oil

    Ćirin-Novta Vera S.; Kevrešan Slavko E.; Kuhajda Ksenija N.; Kandrač Julijan E.; Radić Ljubica M.; Rodić Petar A.

    2003-01-01

    Petroleum acids can be obtained from oil and oil derivatives by alkaline extraction. The aim of this work was to optimize the process of alkaline extraction of petroleum acids from light commercial oil fractions of Vojvodina "Velebit" oil by changing the contact time of oil fraction with strong alkali and by changing the reaction temperature. Optimal conditions for isolation of petroleum acids have been determined to be 9 hours of contact time and reaction temperature of 90°C. Under these con...

  12. Characterization of a retinoic acid responsive element isolated by whole genome PCR.

    Costa-Giomi, M P; Gaub, M P; Chambon, P; Abarzúa, P

    1992-01-01

    We have used whole PCR in an attempt to isolate novel retinoic acid (RA) responsive genes. We cloned several small genomic fragments from total human DNA containing putative retinoic acid responsive elements (RAREs) selected by direct binding to the retinoic acid receptor alpha (RAR alpha). We report here that an oligonucleotide containing a sequence from one of the cloned human DNA fragments, and referred to as alpha 1, functions as an authentic RARE. It is shown that both RAR alpha and RAR ...

  13. Biodistribution in normal mice of an 111In-labelled prostatic acid phosphatase-specific antibody and its F(ab')2 fragments derivatized site-specifically or via bicyclic diethylenetriaminepentaacetic acid anhydride

    We examined the optimization of derivatization of monoclonal antibodies and their fragments intended for use as radiopharmaceutical in radioimaging and/or radioimmunotherapy of prostatic cancer. Two different principles were used to conjugate (DTPA) to a monoclonal antibody (Mab, subclass IgG1) raised against human prostatic acid phosphatase (PAP). In addition, the F(ab')2 fragments of this Mab were also derivatized. The biodistribution of the 111In-labelled derivatives was investigated in normal mice. All the derivatives of IgG1 demonstrated a slower blood clearance than the corresponding derivatives of the F(ab')2 fragments. This property was particularly pronounced in the site-specifically conjugated derivatives of IgG1. All the derivatives studied accumulated in the liver, kidney, and spleen. The CA-DTPA derivatives of F(ab')2 fragments showed the highest kidney-to-blood ratios of radioactivity. The derivatives of IgG1 showed a higher percentage of the injected dose in liver and spleen tissues than the derivatives of the F(ab')2 fragments. The F(ab')2 fragments studied also gave rise to site-specific derivatives, which demonstrated that carbohydrates were also present in this part of the molecule. They behaved similarly to the CA-DTPA F(ab')2 derivative in other respects, but the kidney accumulation was lower at 72 and 120 h. The F(ab')2 fragments studied would be better suited for radioimaging than the derivatives of the IgG1 studied. In contrast, the derivatives of IgG1, especially the p-NH2-Bz-DTPA conjugate, might be more suitable candidates for the development of therapeutic agents. (orig.)

  14. Cdc14 phosphatase

    Machín, Félix; Quevedo Rodriguez, Oliver; Ramos-Pérez, Cristina;

    2016-01-01

    Cycling events in nature start and end to restart again and again. In the cell cycle, whose purpose is to become two where there was only one, cyclin-dependent kinases (CDKs) are the beginning and, therefore, phosphatases must play a role in the ending. Since CDKs are drivers of the cell cycle...

  15. Isolation and characterization of halophilic lactic acid bacteria isolated from "terasi" shrimp paste: a traditional fermented seafood product in Indonesia.

    Kobayashi, Takeshi; Kajiwara, Michika; Wahyuni, Mita; Kitakado, Toshihide; Hamada-Sato, Naoko; Imada, Chiaki; Watanabe, Etsuo

    2003-10-01

    Lactic acid bacteria from "terasi" shrimp paste, a highly popular fermented seafood in Indonesia were isolated and characterized. Viable cell counts were 10(4) to 10(6) cfu/g on MRS medium. All the isolates were catalase-negative, gram-positive cocci and were able to grow at 15% NaCl. Numerical phenotypic analysis showed that the isolates clustered into one group. However, they could be classified into two types: the Tetragenococcus halophilus group and the T. muriaticus group as revealed by a restriction fragment length polymorphism (RFLP) analysis and sequencing of the 16S rRNA gene. This study is the first to show that both species of Tetragenococcus are distributed in Indonesian fermented foods. PMID:14673751

  16. Isolation and cellular fatty acid composition of psychrotrophic Halomonas strains from Antarctic sea water

    Vipra Vijay Jadhav

    2013-06-01

    Full Text Available Microorganisms from extreme environments such as Arctic, Antarctic and Polar regions modulate their membrane fatty acids to survive in such habitats. Characterization of such microorganisms helps in understanding their physiological behavior. In view of this, the present article describes isolation, characterization and cellular fatty acid composition of three bacterial isolates from Antarctic sea water samples. All the three isolates (BRI 6, 29 and 31 were psychrotrophic Gram negative rods. Their 16S rRNA sequencing (around 1200 bp revealed that all three of them belong to genus Halomonas. Each of them showed 99% sequence similarity to Halomonas neptunia Eplume1 (NR 027218, H. boliviensis LC1 (NR 029080 and H. variabilis DSM 3051 (NR 042068. The fatty acid analysis of our isolates indicated i predominance of C 18:1, C 16:0 and C16:1 fatty acids and absence of trans fatty acids in all of them and ii higher percentage of anteiso fatty acids than of iso fatty acids in BRI 6. These characteristic features may contribute to their adaptation to the Antarctic habitat.

  17. Wpływ nawożenia mineralnego i nawadniania na aktyumość peroksydazy, katalazy i fosfatazy kwaśnej w dwóch fazach wzrostu kapusty i porów [The influence of mineral fertilization and irrigation on the activity of peroxidase, catalase and acid phosphatase of cabbage and leek in two stages of growth

    E. Gurgul

    2015-06-01

    Full Text Available During the growth of plant, the very distinct increase of enzymatic activity of peroxidase and catalase was observed, but in case of acid phosphatase in smaller degree. An irrigation caused the decreasing of activity of all tested enzymes in both stages of cabbage growth. However, in case of leeks leaves sprinkling irrigation stimulated activity of catalase and acid phosphatase in both stages and peroxidase in the second stage of growth. The effectiveness of the mineral nutritive was differentiated, and often correlated with a level of soil moisture, kind of plant and its stage of growth.

  18. Effect of andrographolide on phosphatases activity and cytotoxicity against Spodoptera litura

    E Edwin

    2016-05-01

    Full Text Available Andrographolide was isolated from ethanol extraction of Andrographis paniculata by column chromatography. Evaluation of larvicidal efficacy, enzymatic changes and cytotoxic activities against Spodoptera litura were conducted across a range of concentrations. The compound showed significant larvicidal activity between 5 - 25 ppm, post ingestion. Morphological deformities observed in larval-pupal stages. Enzymatic profiles were altered by reduction in acid phosphatase, ACP activity by 69.18 %, alkaline phosphatase, ALP activity 75.3 % and 74.9 % reduction in ATPase. Binding affinity to midgut epithelium cells suggests disintegration of cellular organelles observed was directly associated with ingestion of the compound. The results suggest that andrographolide has potential for development as a significant inhibitor of development against the pest Spodoptera litura.

  19. Induction of nodD Gene in a Betarhizobium Isolate, Cupriavidus sp. of Mimosa pudica, by Root Nodule Phenolic Acids.

    Mandal, Santi M; Chakraborty, Dipjyoti; Dutta, Suhrid R; Ghosh, Ananta K; Pati, Bikas R; Korpole, Suresh; Paul, Debarati

    2016-06-01

    A range of phenolic acids, viz., p-coumaric acid, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid, protocatechuic acid, caffeic acid, ferulic acid, and cinnamic acid have been isolated and identified by LC-MS analysis in the roots and root nodules of Mimosa pudica. The effects of identified phenolic acids on the regulation of nodulation (nod) genes have been evaluated in a betarhizobium isolate of M. pudica root nodule. Protocatechuic acid and p-hydroxybenzoic acid were most effective in inducing nod gene, whereas caffeic acid had no significant effect. Phenylalanine ammonia lyase, peroxidase, and polyphenol oxidase activities were estimated, indicating regulation and metabolism of phenolic acids in root nodules. These results showed that nodD gene expression of betarhizobium is regulated by simple phenolic acids such as protocatechuic acid and p-hydroxybenzoic acid present in host root nodule and sustains nodule organogenesis. PMID:26897126

  20. Isolation of green coffee chlorogenic acids using activated carbon

    Suarez-Quiroz, Mirna Leonor; Alonso Campos, Angelina; Alfaro, Gerardo Valerio; Gonzalez-Rıos, Oscar; Villeneuve, Pierre

    2014-01-01

    Chlorogenic acids, which are interesting natural antioxidants widespread in the plant kingdom, were extracted and purified from Mexican green coffee beans (Coffea arabica) using different methods. The final objective was to find an easy way to extract high-value molecules from a complex mixture, avoiding as much as possible the use of toxic solvents. Three extraction methods (hot water at 80 8C, aqueous methanol 70% (v/v), and aqueous isopropanol 60% (v/v)) were tested in combination with two...

  1. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification

    Mauk, Michael G.; Changchun Liu; Jinzhao Song; Bau, Haim H

    2015-01-01

    Microfluidic components and systems for rapid (<60 min), low-cost, convenient, field-deployable sequence-specific nucleic acid-based amplification tests (NAATs) are described. A microfluidic point-of-care (POC) diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of “lab on a chip” NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (poly...

  2. Poly(N-vinylimidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids.

    Schemeth, Dieter; Noël, Jean-Christophe; Jakschitz, Thomas; Rainer, Matthias; Tessadri, Richard; Huck, Christian W; Bonn, Günther K

    2015-07-23

    In this study we report the novel polymeric resin poly(N-vinyl imidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids. The monomer to crosslinker ratio and the porogen composition were optimized for isolating phenolic acids diluted in acetonitrile at normal phase chromatography conditions, first. Acetonitrile serves as polar, aprotic solvent, dissolving phenolic acids but not interrupting interactions with the stationary phase due to the approved Hansen solubility parameters. The optimized resin demonstrated high loading capacities and adsorption abilities particularly for phenolic acids in both, acetonitrile and aqueous solutions. The adsorption behavior of aqueous standards can be attributed to ion exchange effects due to electrostatic interactions between protonated imidazole residues and deprotonated phenolic acids. Furthermore, adsorption experiments and subsequent curve fittings provide information of maximum loading capacities of single standards according to the Langmuir adsorption model. Recovery studies of the optimized polymer in the normal-phase and ion-exchange mode illustrate the powerful isolation properties for phenolic acids and are comparable or even better than typical, commercially available solid phase extraction materials. In order to prove the applicability, a highly complex extract of rosemary leaves was purified by poly(N-vinyl imidazole/ethylene glycol dimethacrylate) and the isolated compounds were identified using UHPLC-qTOF-MS. PMID:26231906

  3. Isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina L.)

    Saaby, Lasse; Jäger, Anna Katharina; Moesby, Lise;

    2011-01-01

    A previously published systematic review and a metaanalysis have concluded that the consumption of standardized rose hip powder (Rosa canina L.) can reduce pain in osteoarthritis patients. Synovial inflammation has been suggested to play an important role in the pathogenesis of osteoarthritis...... and mainly to involve infiltration of the synovial membrane by macrophages. Therefore, the immunomodulatory effect of standardized rose hip powder of Rosa canina L. was investigated and active principles isolated using the Mono Mac 6 cell line as a model for human macrophages. Treatment of Mono Mac 6 cells...... triterpene acids; oleanolic acid, betulinic acid and ursolic acid (IC(50) 21 +/- 6 microm). Further studies revealed that only oleanolic acid and ursolic acid, but not betulinic acid, could inhibit the lipopolysaccharide induced interleukin-6 release from Mono Mac 6 cells when tested separately. Combination...

  4. Characterization of Lactic Acid Bacteria Isolated from Sauce-type Kimchi.

    Jung, Suk Hee; Park, Joung Whan; Cho, Il Jae; Lee, Nam Keun; Yeo, In-Cheol; Kim, Byung Yong; Kim, Hye Kyung; Hahm, Young Tae

    2012-09-01

    This study was carried out to investigate the isolation and characterization of lactic acid bacteria (LAB) from naturally fermented sauce-type kimchi. Sauce-type kimchi was prepared with fresh, chopped ingredients (Korean cabbage, radish, garlic, ginger, green onion, and red pepper). The two isolated bacteria from sauce-type kimchi were identified as Pediococcus pentosaceus and Lactobacillus brevis by 16S rDNA sequencing and tentatively named Pediococcus sp. IJ-K1 and Lactobacillus sp. IJ-K2, respectively. Pediococcus sp. IJ-K1 was isolated from the early and middle fermentation stages of sauce-type kimchi whereas Lactobacillus sp. IJ-K2 was isolated from the late fermentation stage. The resistance of Pediococcus sp. IJ-K1 and Lactobacillus sp. IJ-K2 to artificial gastric and bile acids led to bacterial survival rates that were 100% and 84.21%, respectively. PMID:24471087

  5. Effect of growth conditions on expression of the acid phosphatase (cyx-appA) operon and the appY gene, which encodes a transcriptional activator of Escherichia coli

    Brøndsted, Lone; Atlung, Tove

    1996-01-01

    The expression and transcriptional regulation of the Escherichia coli cyx-appA operon and the appY gene has been investigated during different environmental conditions using single copy transcriptional lacZ fusions. The cyx-appA operon encodes acid phosphatase and a putative cytochrome oxidase.......ArcA and AppY activated transcription of the cyx-appA operon during entry into stationary phase and under anaerobic growth conditions. The expression of the cyx-appA operon was affected by the anaerobic energy metabolism.The presence of the electron acceptors nitrate and fumarate repressed the expression...... of the cyx-appA operon. The nitrate repression was partially dependent on NarL. A high expression of the operon was obtained in glucose medium supplemented with formate, where E.coli obtains energy by fermentation. The formate induction was independent of the fhlA gene product. The results presented...

  6. Presence of multiple acid phosphatases activity in seedlings of cucumber, radish and rocket salad Presença de atividade de múltiplas fosfatases ácidas em plântulas de pepino, rabanete e rúcula

    Luciane Almeri Tabaldi

    2008-06-01

    Full Text Available Acid phosphatases (3.1.3.2 are a group of enzymes widely distributed in nature, which catalyze the hydrolysis of a variety of phosphate esters in the pH range of 4-6. We confirmed the presence of acid phosphatases in seedlings of cucumber (Cucumis sativus, radish (Raphanus sativus and rocket salad (Eruca vesicaria under different assay conditions using a rapid and simple preparation. The results showed that the optimum pH and temperature used for all species were close to 5.5 and 35°C, respectively. The enzyme was inhibited by molybdate, fluoride, azide, levamisole, orthovanadate, Zn2+ and Cu2+. Suramin had no effect on enzyme activity. The acid phosphatase from cucumber, radish and rocket salad hydrolyzed a wide variety of phosphate esters and the highest activity was observed with PPi, ATP and GTP. These results demonstrate that the enzyme investigated in this study is different from well known ester phosphate cleaving plant enzymes (apyrase and inorganic pyrophosphatases and this preparation could be a useful tool to future toxicological studies and to study initially all isoforms of acid phosphatase.As fosfatases ácidas (3.1.3.2 são um grupo de enzimas amplamente distribuídas na natureza, as quais catalisam a hidrólise de uma variedade de ésteres de fosfato com uma variação de pH entre quatro e seis. Foi confirmada a presença de fosfatases ácidas em plântulas de pepino (Cucumis sativus, rabanete (Raphanus sativus e rúcula (Eruca vesicaria sob diferentes condições de ensaio usando uma preparação rápida e simples. Os resultados mostraram que o pH e a temperatura ótimos para todas as espécies foram 5,5 e 35°C, respectivamente. A enzima foi inibida por molibdato, fluoreto, azida, levamisole, ortovanadato, Zn2+ e Cu2+. O inibidor suramim não afetou a atividade enzimática. As fosfatases ácidas de pepino, rabanete e rúcula hidrolisaram uma ampla variedade de ésteres de fosfato e a maior atividade foi observada com PPi, ATP

  7. Isolation from Cussonia barteri of 1'-O-chlorogenoylchlorogenic acid and 1'-O-chlorogenoylneochlorogenic acid, a new type of quinic acid esters.

    Papajewski, S; Vogler, B; Conrad, J; Klaiber, I; Roos, G; Walter, C U; Süssmuth, R; Kraus, W

    2001-11-01

    1'-O-Chlorogenoylchlorogenic acid and 1'-O-chlorogenoylneochlorogenic acid, a new type of quinic acid esters, have been isolated, in addition to six known quinic acid esters, rutin, and a mixture of saponins, from the methanol extract of Cussonia barteri Seemann (Araliaceae) leaves collected in Cameroon. Structure determination was achieved by NMR, mass, IR, and UV spectroscopy. All compounds were tested for inhibitory activity on 5-lipoxygenase and cyclooxygenase-1, for antimicrobial activity against Bacillus subtilis, Pseudomonas fluorescens, and Cladosporium cucumerinum, and for haemolytic activity. PMID:11731915

  8. Mycophenolic acid inhibits replication of Type 2 Winnipeg, a cerebrospinal fluid-derived reovirus isolate

    Hermann, Laura L.; Coombs, Kevin M.

    2004-01-01

    BACKGROUND: The role of reoviruses in human disease is uncertain. Most identified cases are sporadic and asymptomatic or produce minor upper respiratory or gastrointestinal symptoms. In November 1997, a reovirus was isolated from the cerebrospinal fluid of a severe combined immune deficient infant in Winnipeg, Manitoba. RNA characterization and sequencing studies demonstrated this reovirus isolate to be unique. Thus, the virus was named Type 2 Winnipeg (T2W).OBJECTIVES: Mycophenolic acid (MPA...

  9. Characterization and in vitro probiotic evaluation of lactic acid bacteria isolated from idli batter

    Iyer, Bharti K.; Singhal, Rekha S.; Ananthanarayan, Laxmi

    2011-01-01

    An Indian traditional fermented food, idli batter, was used as a source for isolation of lactic acid bacteria (LAB). A total of 15 LAB strains were isolated on the basis of their Gram nature and catalase activity. Of these, one lactobacilli strain and one lactococci strain which showed antimicrobial activity were identified using biochemical characterization, sugar utilization and molecular sequencing. The microbes, labeled as IB-1 (Lactobacillus plantarum) and IB-2 (Lactococcus lactis) were ...

  10. Investigation of antibacterial, acid and bile tolerance properties of lactobacilli isolated from Koozeh cheese

    Hassan Hassanzadazar

    2012-09-01

    Full Text Available Lactobacillus strains are a major part of the probiotics, microflora of the intestine and of fermented dairy products, and are found in a variety of environments. The aim of this study was to find out the ability of bile and acid tolerance and antibacterial properties of the twenty eight isolates of three group lactobacilli namely Lactobacillus plantarum, Lactobacillus casei and Lactobacillus delbruki. For this purpose Twenty eight different Lactobacillus strains that isolated from Koozeh cheese as a traditional cheese were screened. The acid tolerance test was studied under pH 2.0 and 3.0 with 7.5 as control. The cell count for the acid tolerance test was obtained at an interval of 0, 1, 2 and 3 hours respectively and was pour plated on Man, Rogosa, and Sharpe (MRS agar to be incubated at 37 °C for 24 hours. All cells were selected for bile tolerance test in MRS broth containing bile concentrations of 0% as control and 0.3% as test. Then cell counts were enumerated after 24 hours of incubation on MRS agar. Results showed twenty seven isolates did not have ability to tolerate acid and bile salts and antimicrobial activity against four indicator bacteria included Eshirichia coli, Listeria monocytogenesis, bacillus cereus, Salmonella entritidis. Only one Isolate namely Lactobacillus casei could tolerate acid and bile salt and had antibacterial activity against of L. monocytogenesis. Therefore we can consider this strain as a native probiotic but extra examinations was required.

  11. A novel protein tyrosine phosphatase 1B inhibitor from Tinospora sinensis

    Prasoon Gupta; Upasana Sharma; Gupta, Praveen K.; Rakesh Maurya

    2012-01-01

    Bioassay-directed fractionation led to the identification of a new compound, 4-hydroxy-heptadec-6-enoic acid ethyl ester (1) together with three known compounds (2-4) from Tinospora sinensis. The structure of 1 was determined by analysis of spectroscopic data. The isolated compounds were evaluated for their protein tyrosine phosphatase 1B (PTP1B) inhibitory activity. Compounds 1 and 2 displayed significant inhibitory activity with IC 50 values 61.1 and 74.2 μM, respectively

  12. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Ghabbour, N.; Lamzira, Z.; Thonart, P.; Cidalia, P.; Markaouid, M.; Asehraoua, A.

    2011-07-01

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo-4-chloro-3-indolyl {beta}-D-glucuronide (X-Gluc) as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis. (Author).

  13. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo-4-chloro-3-indolyl β-D-glucuronide (X-Gluc) as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis. (Author).

  14. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Ghabbour, N.; Lamzira, Z.; Thonart, P.; Cidalia, P.; Markaoui, M.; Asehraou, A.

    2011-01-01

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade ole...

  15. Isolation and characterization of hyaluronic acid from the liver of marine stingray Aetobatus narinari.

    Sadhasivam, Giji; Muthuvel, Arumugam; Pachaiyappan, Abirami; Thangavel, Balasubramanian

    2013-03-01

    Although hyaluronic acid research pursuits ahead in exploring its biomedical perspective, very limited investigations were carried out in their isolation shape view point, furthermore, most of the investigations were targeted towards the terrestrial source. To swerve from that, the present study was projected through the marine superstore, where in high molecular weight hyaluronic acid of 13, 65,863 Da was isolated from the liver of stingray Aetobatus narinari. The purified HA was confirmed at the preliminary level by their stains all dye binding nature. Their analytical composition including carbon, hydrogen, nitrogen, N-acetyl glucosamine, glucuronic acid contents was analysed. The HA was characterized by agarose-gel electrophoresis, FTIR, HPTLC, and (1)H NMR. The DPPH radical scavenging activity of HA and its reducing power was evident to all the tested concentrations, but lower than that of ascorbic acid. HA showed significant inhibition against the proliferation of cells, substantiating its influence in regulation of cell functions. PMID:23220595

  16. Cellular fatty acid composition, protein profile and antimicrobial activity of Bacillus sp., isolated from fish gut

    Pushparaj Sujith; Baskaran Rohini; Singaram Jayalakshmi

    2014-01-01

    Objective: To purify and partially characterize the antimicrobial compounds from bacteriaBacillus sp., isolated from fish gut. Methods: Protein and fatty acids were isolated from the bacteria and checked for the presence of antibacterial activity. Protein has been purified to apparent homogeneity from the supernatants of culture by means of ammonium sulphate precipitation followed by dialysis. Fourier transform infrared spectroscopy analyses were performed for proteins to identify the functional groups.Results:sulfate polyacrylamide gel electrophoresis. Fatty acids were extracted and subjected to gas chromatographic analysis.Conclusions:Protein showed an apparent molecular mass 56, 47 and 39 kDa on sodium dodecyl acids and proteins which holds promise for the development of new drugs. The antimicrobial activity of the bacteria might be due to the presence of fatty acids and proteins which holds promise for the development of new drugs.

  17. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification

    Michael G. Mauk

    2015-10-01

    Full Text Available Microfluidic components and systems for rapid (<60 min, low-cost, convenient, field-deployable sequence-specific nucleic acid-based amplification tests (NAATs are described. A microfluidic point-of-care (POC diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of “lab on a chip” NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (polymerase-chain reaction-based tests in clinical laboratories can be realized with a disposable, palm-sized, plastic microfluidic chip in which: (1 nucleic acids (NAs are extracted from relatively large (~mL volume sample lysates using an embedded porous silica glass fiber or cellulose binding phase (“membrane” to capture sample NAs in a flow-through, filtration mode; (2 NAs captured on the membrane are isothermally (~65 °C amplified; (3 amplicon production is monitored by real-time fluorescence detection, such as with a smartphone CCD camera serving as a low-cost detector; and (4 paraffin-encapsulated, lyophilized reagents for temperature-activated release are pre-stored in the chip. Limits of Detection (LOD better than 103 virons/sample can be achieved. A modified chip with conduits hosting a diffusion-mode amplification process provides a simple visual indicator to readily quantify sample NA template. In addition, a companion microfluidic device for extracting plasma from whole blood without a centrifuge, generating cell-free plasma for chip-based molecular diagnostics, is described. Extensions to a myriad of related applications including, for example, food testing, cancer screening, and insect genotyping are briefly surveyed.

  18. Isolation and characterization of Halomonas sp strain IMPC, a p-coumaric acid-metabolizing bacterium that decarboxylates other cinnamic acids under hypersaline conditions

    Abdelkafi, Slim; Labat, Marc; Casalot, Laurence; Chamkha, M.; Sayadi, S.

    2006-01-01

    A moderately halophilic, mesophilic, Gram-negative, motile, nonsporulating bacterium, designated strain IMPC, was isolated from a table-olive fermentation rich in aromatic compounds, after enrichment on p-coumaric acid under halophilic conditions. Strain IMPC was able to degrade p-coumaric acid. p-hydroxybenzaldehyde and p-hydroxybenzoic acid were detected as breakdown products from p-coumaric acid. Protocatechuic acid was identified as the final aromatic product of p-coumaric acid catabolism...

  19. The aflatoxin B1 isolating potential of two lactic acid bacteria

    Adel Hamidi; Reza Mirnejad; Emad Yahaghi; Vahid Behnod; Ali Mirhosseini; Sajad Amani; Sara Sattari; Ebrahim Khodaverdi Darian

    2013-01-01

    Objective:To determine lactic acid bacteria’s capability to enhance the process of binding and isolating aflatoxin B1 and to utilize such lactic acid bacteria as a food supplement or probiotic products for preventing absorption of aflatoxin B1 in human and animal bodies. Methods: In the present research, the bacteria were isolated from five different sources. For surveying the capability of the bacteria in isolating aflatoxin B1, ELISA method was implemented, and for identifying the resultant strains through 16S rRNA sequencing method, universal primers were applied. Results: Among the strains which were isolated, two strains of Lactobacillus pentosus and Lactobacillus beveris exhibited the capability of absorbing and isolating aflatoxin B1 by respectively absorbing and discharging 17.4%and 34.7%of the aforementioned toxin existing in the experiment solution. Conclusions:Strains of Lactobacillus pentosus and Lactobacillus beveris were isolated from human feces and local milk samples, respectively. And both strains has the ability to isolate or bind with aflatoxin B1.

  20. ISOLATION AND IDENTIFICATION OF LACTIC ACID PRODUCING BACTERIA FROM CAMEL MILK

    Toqeer Ahmad, Rashida Kanwal, Izhar Hussain Athar1, Najam Ayub

    2002-01-01

    Lactic acid bacteria (LAB) were isolated from camel milk by culturing the camel milk on specific media and pure culture was obtained by sub culturing. Purification of culture was confirmed by Gram's staining and identified by different bio-chemical tests. Camel milk contains lactic acid producing bacteria including Strpptococci such as S. cremoris and S. lactis and Lactobacilli such as L. acidophilus L. acidophilus grows more rapidly in camel milk than others as its growth is supported by cam...

  1. Initial performance of corn in response to treatment of seeds with humic acids isolated from bokashi

    Marihus Altoé Baldotto; Lílian Estrela Borges Baldotto

    2016-01-01

    ABSTRACT The humified organic matter presents bioactivity similar to the auxinic effect. As bokashi is produced by a special process of humification, information is needed about the bioactive potential of its humic acids. The objective of this work was studying the initial performance of corn-indicator plants in response to the application of different concentrations of humic acids isolated from bokashi. The corn seeds were treated for 16 hours with solutions containing 0, 10, 20, 30, 40 and ...

  2. Selection of exopolysaccharide-producing lactic acid bacteria isolates from Inner Mongolian traditional yoghurt

    Zhang Chun-lei; Li Jia-qi; Guo Hai-tao; Wang Jie; Xu Ri-hua

    2014-01-01

    Lactic acid bacteria (LAB) isolated from Inner Mongolian traditional yoghurt were evaluated for the production of exopolysaccharides (EPS) by phenol-sulphuric acid method after ethanol precipitation and dialysis. Total polysaccharide was extracted from sucrose-containing MRS broth cultures of the selected LAB strains. Comparison of the EPS yields revealed that among tested LAB, strain 37 exhibited the highest production of 536.904 mg/L. The strain was identified as Leuconostoc citreum with ca...

  3. Unambiguous typing of canine adenovirus isolates by deoxyribonucleic acid restriction-endonuclease analysis.

    Assaf, R; Marsolais, G; Yelle, J; Hamelin, C

    1983-01-01

    Viral deoxyribonucleic acid extracted from a limited number of cells infected with canine adenovirus type 1 or type 2 was cleaved with several restriction endonucleases. Agarose gel electrophoresis of the limit digests showed stable differences between the canine adenovirus type 1 and type 2 cleavage patterns. Rapid and accurate typing of large numbers of clinical isolates may thus be done by deoxyribonucleic acid restriction-endonuclease analysis.

  4. Isolation and characterization of lactic acid bacteria and yeasts from the Brazilian grape sourdough

    Krischina Singer Aplevicz; Jaciara Zarpellon Mazo; Eunice Cassanego Ilha; Andréia Zilio Dinon; Ernani Sebastião Sant´Anna

    2014-01-01

    Sourdough is a mixture of flour and water fermented by lactic acid bacteria and yeast, with a large use in bakery products. This study was developed with Brazilian grape (Niagara rosada) sourdough obtained from spontaneous fermentation. The aim of this work was to characterize genotypic and phenotypically lactic acid bacteria and yeasts isolated from sourdough. The phenotypic identification for bacteria and yeasts was performed by using the kit API50CHL and 20CAUX and the genotypic characteri...

  5. Isolation of 14C labelled amino acids by biosynthesis in maize plants (Zea mais L.)

    A method of obtaining 14C labelled amino acids by biosynthesis in maize plants which had assimilated 14CO2, has been assayed. The plants were labelled for 60 minutes with 14CO2 produced from Ba 14CO3 (specific activity of 148 KBq/μmol). An extract of the soluble compounds was obtained with 80% ethanol and the amino acids were separated from the rest of the soluble compounds by ion exchange chromatography on column of Dowex 50-X8 resin. Finally, seventeen amino acids were isolated and identified from the purified extract. The acid amino acids were separated in anionic column (Dowex 1-X8) and the neutral and basic amino acids in cationic column (Dowex 50-X4). (Author) 56 refs

  6. FISH to meiotic pachytene chromosomes of tomato locates the root-knot nematode resistance gene Mi-1 and the acid phosphatase gene Aps-1 near the junction of euchromatin and pericentromeric heterochromatin of chromosome arms 6S and 6L, respectively

    Zhong, X.B.; Bodeau, J.; Fransz, P.F.; Williamson, V.M.; Kammen, van A.; Jong, de J.H.; Zabel, P.

    1999-01-01

    The root-knot nematode resistance gene Mi-1 in tomato has long been thought to be located in the pericentromeric heterochromatin region of the long arm of chromosome 6 because of its very tight genetic linkage (approx. 1 cM) to the markers Aps-1 (Acid phosphatase 1) and yv (yellow virescent). Using

  7. Isolation of the stable fraction (the core) of the humic acid.

    Adani, Fabrizio; Ricca, Giuliana; Tambone, Fulvia; Genevini, Pierluigi

    2006-11-01

    Humic acid consists of a recalcitrant (unhydrolysed fraction) (the core) and labile (hydrolysable fraction) fraction. Core-humic acid (core-HA) isolation was performed by treating source material with apolar and polar solvents (organic solvents+acid hydrolysis) before alkaline extraction. Leonardite, soil Ah horizont and dry blood were chosen for this study because of their different origin and degree of humification. Chemical analysis (elemental analysis, total acidity, E(4):E(6)), spectroscopic analysis (DRIFT and (1)H NMR), and complete mass balance were used to investigate the effect of purifying humic acids. The results obtained showed that purification produced a slight modification of Leonardite humic acids as was expected for these highly humified organic matrices. On the other hand, about 500 g kg(-1) of soil humic acids were lost by purification. The fractions lost mainly consisted of carbohydrates. Dry blood showed the presence of humic acids that contrasted with its origin, thus indicating the limitations of the common analytical methods used for HA extraction. Nevertheless, in practice, purification caused the complete disappearance (914 g kg(-1) of HA was lost) of these HAs. The results obtained in this work suggest that the HA fraction isolated (named core-HA) effectively represents the HA structure proposed by the existing literature, since the purification proposed was able to eliminate the adsorbed organic molecules (interference materials) coating the HA structure. PMID:16735055

  8. Acid-base and copper-binding properties of three organic matter fractions isolated from a forest floor soil solution

    van Schaik, Joris W. J.; Kleja, Dan B.; Gustafsson, Jon Petter

    2010-01-01

    Vast amounts of knowledge about the proton- and metal-binding properties of dissolved organic matter (DOM) in natural waters have been obtained in studies on isolated humic and fulvic (hydrophobic) acids. Although macromolecular hydrophilic acids normally make up about one-third of DOM, their proton- and metal-binding properties are poorly known. Here, we investigated the acid-base and Cu-binding properties of the hydrophobic (fulvic) acid fraction and two hydrophilic fractions isolated from ...

  9. Poly(N-vinylimidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids

    Highlights: • Free-radical polymerization of protonable vinylimidazole with EGMDA. • Polymer-optimization by maximum loading capacity of phenolic acids. • Performs better than SiO2 and Al2O3 in normal phase mode using acetonitrile. • Performs equal or even better in anion-exchange mode compared to Oasis-MAX. • Efficient purification of phenolic compounds from crude extract. - Abstract: In this study we report the novel polymeric resin poly(N-vinyl imidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids. The monomer to crosslinker ratio and the porogen composition were optimized for isolating phenolic acids diluted in acetonitrile at normal phase chromatography conditions, first. Acetonitrile serves as polar, aprotic solvent, dissolving phenolic acids but not interrupting interactions with the stationary phase due to the approved Hansen solubility parameters. The optimized resin demonstrated high loading capacities and adsorption abilities particularly for phenolic acids in both, acetonitrile and aqueous solutions. The adsorption behavior of aqueous standards can be attributed to ion exchange effects due to electrostatic interactions between protonated imidazole residues and deprotonated phenolic acids. Furthermore, adsorption experiments and subsequent curve fittings provide information of maximum loading capacities of single standards according to the Langmuir adsorption model. Recovery studies of the optimized polymer in the normal-phase and ion-exchange mode illustrate the powerful isolation properties for phenolic acids and are comparable or even better than typical, commercially available solid phase extraction materials. In order to prove the applicability, a highly complex extract of rosemary leaves was purified by poly(N-vinyl imidazole/ethylene glycol dimethacrylate) and the isolated compounds were identified using UHPLC–qTOF-MS

  10. Poly(N-vinylimidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids

    Schemeth, Dieter; Noël, Jean-Christophe [Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, CCB—Center of Chemistry and Biomedicine, Innrain 80-82, 6020 Innsbruck (Austria); Jakschitz, Thomas [Austrian Drug Screening Institute, Innrain 66a, 6020 Innsbruck (Austria); Rainer, Matthias, E-mail: m.rainer@uibk.ac.at [Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, CCB—Center of Chemistry and Biomedicine, Innrain 80-82, 6020 Innsbruck (Austria); Tessadri, Richard [Institute of Mineralogy and Petrography, Leopold-Franzens University of Innsbruck, Innrain 52, 6020 Innsbruck (Austria); Huck, Christian W. [Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, CCB—Center of Chemistry and Biomedicine, Innrain 80-82, 6020 Innsbruck (Austria); Bonn, Günther K. [Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, CCB—Center of Chemistry and Biomedicine, Innrain 80-82, 6020 Innsbruck (Austria); Austrian Drug Screening Institute, Innrain 66a, 6020 Innsbruck (Austria)

    2015-07-23

    Highlights: • Free-radical polymerization of protonable vinylimidazole with EGMDA. • Polymer-optimization by maximum loading capacity of phenolic acids. • Performs better than SiO{sub 2} and Al{sub 2}O{sub 3} in normal phase mode using acetonitrile. • Performs equal or even better in anion-exchange mode compared to Oasis-MAX. • Efficient purification of phenolic compounds from crude extract. - Abstract: In this study we report the novel polymeric resin poly(N-vinyl imidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids. The monomer to crosslinker ratio and the porogen composition were optimized for isolating phenolic acids diluted in acetonitrile at normal phase chromatography conditions, first. Acetonitrile serves as polar, aprotic solvent, dissolving phenolic acids but not interrupting interactions with the stationary phase due to the approved Hansen solubility parameters. The optimized resin demonstrated high loading capacities and adsorption abilities particularly for phenolic acids in both, acetonitrile and aqueous solutions. The adsorption behavior of aqueous standards can be attributed to ion exchange effects due to electrostatic interactions between protonated imidazole residues and deprotonated phenolic acids. Furthermore, adsorption experiments and subsequent curve fittings provide information of maximum loading capacities of single standards according to the Langmuir adsorption model. Recovery studies of the optimized polymer in the normal-phase and ion-exchange mode illustrate the powerful isolation properties for phenolic acids and are comparable or even better than typical, commercially available solid phase extraction materials. In order to prove the applicability, a highly complex extract of rosemary leaves was purified by poly(N-vinyl imidazole/ethylene glycol dimethacrylate) and the isolated compounds were identified using UHPLC–qTOF-MS.

  11. Antimicrobial Activity of Cell Free Supernatant of Irradiated Lactic Acid Bacteria Isolates

    Attempts were made to isolate bio preservatives using food wastes with no value and low cost. Whey is the raw material achieved that value. Whey and many other food wastes are used in our study to isolate Lactic acid bacteria (LAB). Cell free supernatants (CFS) of isolates are used to evaluate their antimicrobial activity against indicator pathogenic bacterial strains. CFS-9 isolate from whey has the highest inhibitory activity compared to all other isolates. The inhibitory activity of CFS-9, Nisin (400 IU / ml) and the standard Lactococcus Lactis Subsp. Lactis ATCC 11454 (Lacto) were determined. Furthermore, isolate-9 and Lacto strains were exposed to irradiation at different doses. The inhibition zones of; control isolate-9 (non-irradiated) showed the highest values against all indicator strains, CFS of irradiated Lacto at dose 250 Gy was the highest value against Bacillus cereus and Escherichia coli compared to other irradiation treatments, CFS of irradiated Lacto at dose 100 Gy was the highest value against Staph aureus, while the inhibition zone was in the highest value in CFS of irradiated Lacto at dose 500 Gy against Salmonella typhimurium. Nisin (400 IU / ml) was significantly higher than all CFS of irradiated isolate-9 while, the inhibition zones of all CFS-Lacto (irradiated and nonirradiated) are better and higher than nisin-400

  12. Isolation and characterisation of high molecular weight ( sup 3 H)hyaluronic acid

    Chabrecek, P. (Slovak Technical Univ., Bratislava (Czechoslovakia). Inst. of Biotechnology); Soltes, L.; Kallay, Z. (Slovenska Akademia Vied, Bratislava (Czechoslovakia). Ustav Experimentalnej Farmakologie); Fugedi, A. (Slovenska Akademia, Bratislava (Czechoslovakia). Computing Centre)

    1990-10-01

    A high-performance gel permeation chromatographic separation method was developed for the isolation and characterisation of high molecular weight ({sup 3}H)hyaluronic acid. The molecular characteristics of the labelled sample were M{sub w}=3.92 x 10{sup 5} Da, M{sub w}/M{sub n}=1.55. (author).

  13. Isolation and characterisation of high molecular weight [3H]hyaluronic acid

    A high-performance gel permeation chromatographic separation method was developed for the isolation and characterisation of high molecular weight [3H]hyaluronic acid. The molecular characteristics of the labelled sample were Mw=3.92 x 105 Da, Mw/Mn=1.55. (author)

  14. Characterization of airag collected in Ulaanbaatar, Mongolia with emphasis on isolated lactic acid bacteria

    Choi, Suk-Ho

    2016-01-01

    Background Airag, alcoholic sour-tasting beverage, has been traditionally prepared by Mongolian nomads who naturally ferment fresh mares’ milk. Biochemical and microbiological compositions of airag samples collected in Ulaanbaatar, Mongolia and physiological characteristics of isolated lactic acid bacteria were investigated. Methods Protein composition and biochemical composition were determined using sodium dodecyl sulfate-gel electrophoresis and high performance liquid chromatography, respe...

  15. Diacidene, a polyene dicarboxylic acid from a Micromonospora isolate from the German Wadden Sea.

    Ohlendorf, Birgit; Schulz, Dirk; Beese, Pascal; Erhard, Arlette; Schmaljohann, Rolf; Imhoff, Johannes F

    2012-01-01

    Micromonospora sp. strain DB620 was isolated from a Wadden Sea sediment sample collected near Büsum (Germany) and is closely related (99% 16S-rRNA gene sequence similarity) to Micromonospora coxensis strain MTCC8093. It produced a new polyene dicarboxylic acid named diacidene (1) and in addition a derivative of chorismic acid, the known 3-[(1-carboxyvinyl)oxy]benzoic acid. The structure elucidation of 1 was achieved by applying different 1D and 2D NMR techniques as well as mass spectrometry and UV spectroscopy. PMID:23198401

  16. Citric Acid Production by the Aspergillus niger Isolated from the Microflora of Iran

    R. Yazdanparast; and A. Bahrani

    1995-01-01

    Citric acid production by A.niger, isolated from the microflora of Iran, has been investigated in liquid and semi-solid states using growth media with different compositions. In 2% media made of Rocheh grape pomace or sabouraud dextrose, the yield of citric acid production was 0.7 g per Kg of the pomace; and the yield decreased by 50% in 2% saghal solian grape pomace medium. However, in 40% (W/W) saghal solian semi-solid medium containing 3% methanol, the yield of citric acid production has i...

  17. Isolation and preparation of cytotoxic chromanone acids from the bark of Calophyllum dongnaiense

    Nguyen, Tri Hieu; Hansen, Poul Erik; Duus, Fritz; Heilmann, Gørg; Pham, Dinh Hung; Nguyen, Lien Hoa Dieu

    2009-01-01

    Three chromanone acids, blancoic acid (1), isoblancoic acid (2) and chapelieric acid (3), were isolated from the bark of Calophyllum dongnaiense Pierre collected in Dong Nai Province. Their structures were elucidated using spectroscopic techniques (1-D and 2-D NMR, UV, IR and MS) as well as...

  18. Alkaline Phosphatase in Stem Cells

    Kateřina Štefková

    2015-01-01

    Full Text Available Alkaline phosphatase is an enzyme commonly expressed in almost all living organisms. In humans and other mammals, determinations of the expression and activity of alkaline phosphatase have frequently been used for cell determination in developmental studies and/or within clinical trials. Alkaline phosphatase also seems to be one of the key markers in the identification of pluripotent embryonic stem as well as related cells. However, alkaline phosphatases exist in some isoenzymes and isoforms, which have tissue specific expressions and functions. Here, the role of alkaline phosphatase as a stem cell marker is discussed in detail. First, we briefly summarize contemporary knowledge of mammalian alkaline phosphatases in general. Second, we focus on the known facts of its role in and potential significance for the identification of stem cells.

  19. Antilisterial activity of lactic acid bacteria isolated from vacuum-packaged brazilian meat and meat products

    Martinis Elaine C.P. De

    2001-01-01

    Full Text Available Twenty samples of Brazilian meat and meat products were screened by the agar overlay method for bacteriocin-producing lactic acid bacteria, using Lactobacillus sake ATCC 15521 as indicator strain. Based on Gram staining, KOH reaction, catalase test and fermentation of 49 carbohydrates (API 50 CH, three out of seven isolates with confirmed antagonist properties were identified as Lactobacillus curvatus, one as Leuconostoc mesenteroides and one as Leuconostoc sp. Two isolates could not be properly identified using these tests. The inhibitors produced by these strains were sensitive to proteases. Inhibition due to lytic bacteriophages was ruled out, so the isolates were classified as bacteriocin-producing lactic acid bacteria. Four of them presented antilisterial activity and a potential application as biopreservatives in meat systems.

  20. Direct determination of phosphatase activity from physiological substrates in cells.

    Zhongyuan Ren

    Full Text Available A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min(-1 mg(-1 for PPi, to 56 ± 11 nmol min(-1 mg(-1 for AMP, to 79 ± 23 nmol min(-1 mg(-1 for beta-glycerophosphate and to 73 ± 15 nmol min(-1 mg(-1 for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole--a TNAP inhibitor--served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes.

  1. SELECTED PROPERTIES OF LACTIC ACID BACTERIA ISOLATED FROM RAW COW'S MILK

    Miroslav Kročko

    2012-02-01

    Full Text Available Normal 0 21 false false false MicrosoftInternetExplorer4 For the identification of lactic acid bacteria derived from raw cow's milk, 151 colonies were isolated. The grow conditions of lactic acid bacteria were at temperature 37 °C for 3 days on MRS medium. Based on microscopical preparation, negative catalase and Gram-positive test were 81 isolates confirmed as genus Lactobacillus. Out of these, 9 isolates were evaluated for acidifying activity in UHT milk at 25 °C, 30 °C and 37 °C at regular intervals during 24 hours. The average count of NSLAB lactobacilli in raw cow's milk reached the value 1.54.104 KTJ.ml-1. It was found that all tested strains of lactobacilli did not cause significant changes of titratable acidity in milk at 25 °C and 30 °C. Only one strain significantly improved the titratable acidity of milk at 37 °C after 24 hours. The acidity reached the value from 7.5 °SH to 41.9 °SH. This strain was confirmed by PCR method as Lactobacillus helveticus.doi:10.5219/177

  2. Modulators of intestinal alkaline phosphatase.

    Bobkova, Ekaterina V; Kiffer-Moreira, Tina; Sergienko, Eduard A

    2013-01-01

    Small molecule modulators of phosphatases can lead to clinically useful drugs and serve as invaluable tools to study functional roles of various phosphatases in vivo. Here, we describe lead discovery strategies for identification of inhibitors and activators of intestinal alkaline phosphatases. To identify isozyme-selective inhibitors and activators of the human and mouse intestinal alkaline phosphatases, ultrahigh throughput chemiluminescent assays, utilizing CDP-Star as a substrate, were developed for murine intestinal alkaline phosphatase (mIAP), human intestinal alkaline phosphatase (hIAP), human placental alkaline phosphatase (PLAP), and human tissue-nonspecific alkaline phosphatase (TNAP) isozymes. Using these 1,536-well assays, concurrent HTS screens of the MLSMR library of 323,000 compounds were conducted for human and mouse IAP isozymes monitoring both inhibition and activation. This parallel screening approach led to identification of a novel inhibitory scaffold selective for murine intestinal alkaline phosphatase. SAR efforts based on parallel testing of analogs against different AP isozymes generated a potent inhibitor of the murine IAP with IC50 of 540 nM, at least 65-fold selectivity against human TNAP, and >185 selectivity against human PLAP. PMID:23860652

  3. Antibacterial Activity of Some Lactic Acid Bacteria Isolated from an Algerian Dairy Product

    Abdelkader Mezaini

    2009-01-01

    Full Text Available In the present study, the antibacterial effect of 20 lactic acid bacteria isolates from a traditional cheese was investigated. 6 isolates showed antibacterial effect against Gram positive bacteria. Streptococcus thermophilus T2 strain showed the wide inhibitory spectrum against the Gram positive bacteria. Growth and bacteriocin production profiles showed that the maximal bacteriocin production, by S. thermophilus T2 cells, was measured by the end of the late-log phase (90 AU ml−1 with a bacteriocine production rate of 9.3 (AU ml−1 h−1. In addition, our findings showed that the bacteriocin, produced by S. thermophilus T2, was stable over a wide pH range (4–8; this indicates that such bacteriocin may be useful in acidic as well as nonacidic food. This preliminarily work shows the potential application of autochthonous lactic acid bacteria to improve safety of traditional fermented food.

  4. Cellular fatty acid composition, protein profile and antimicrobial activity of Bacillus sp., isolated from fish gut

    Pushparaj Sujith

    2014-01-01

    Full Text Available Objective: To purify and partially characterize the antimicrobial compounds from bacteria Bacillus sp., isolated from fish gut. Methods: Protein and fatty acids were isolated from the bacteria and checked for the presence of antibacterial activity. Protein has been purified to apparent homogeneity from the supernatants of culture by means of ammonium sulphate precipitation followed by dialysis. Fourier transform infrared spectroscopy analyses were performed for proteins to identify the functional groups. Results: Protein showed an apparent molecular mass 56, 47 and 39 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Fatty acids were extracted and subjected to gas chromatographic analysis. Conclusions: The antimicrobial activity of the bacteria might be due to the presence of fatty acids and proteins which holds promise for the development of new drugs.

  5. Leojaponic acids A and B, two new homologous terpenoids, isolated from Leonurus japonicus.

    Wu, Han-Kui; Mao, Yan-Jun; Sun, Shan-Shan; Xu, Zhi-Yong; Ma, Ya; Cao, Jin-Xia; Qi, He; Wu, Zhi-Fu; Li, Gang; Yang, Wei-Hua

    2016-04-01

    The present study aimed at isolation and purification of the bioactive terpenoids from the herb of Leonurus japonicus by chromatographic separations such as silica gel, sephadex LH-20 and C18 reversed phase silica gel, as well as preparative HPLC. As a result, leojaponic acids A (1, C17H24O4) and B (2, C18H26O4), two homologous terpenoids, together with (-)-loliolide (3), 1-(3-ethylphenyl) ethane-1, 2-diol (4) and dibutyl phthalate (5), were isolated from the EtOH extract of L. japonicus. All the chemical structures of the isolates were elucidated on the basis of 1D and 2D NMR analyses. Compounds 1 and 2 were new terpenoids, and Compounds 3 and 4 were isolated and identified for the first time from this plant. In addition, the α-glucosidase and tyrosinase inhibitory activity of the new compounds were evaluated. PMID:27114319

  6. Characteristics of lactic acid bacteria isolates and their effect on silage fermentation of fruit residues.

    Yang, Jinsong; Tan, Haisheng; Cai, Yimin

    2016-07-01

    The natural lactic acid bacteria (LAB) population, chemical composition, and silage fermentation of fruit residues were studied. Eighty-two strains of LAB were isolated from fruit residues such as banana leaf and stem, pineapple peel, and papaya peel. All strains were gram-positive and catalase-negative bacteria, and they were divided into 7 groups (A-G) according to morphological and biochemical characters. Strains in groups A to F were rods, and group G was cocci. Group F produced gas from glucose; other groups did not. Groups A to C and F formed dl-lactic acid, whereas groups D, E, and G formed l-lactic acid. Based on the 16S rRNA gene sequence and DNA-DNA hybridization analysis, groups A to G strains were identified as Lactobacillus plantarum (54.9% of the total isolates), Lactobacillus paraplantarum (3.6%), Lactobacillus nagelii (8.5%), Lactobacillus perolens (4.9%), Lactobacillus casei (11.0%), Lactobacillus fermentum (9.8%), and Enterococcus gallinarum (7.3%), respectively. Lactobacillus plantarum and Lactobacillus casei are the most frequently isolated from fruit residues as a dominant species, and they could grow at a lower pH conditions and produce more lactic acid than other isolates. Pineapple and papaya peels contained higher crude protein (11.5-13.8%) and water-soluble carbohydrate (16.8-22.4%), but lower acid detergent fiber contents (21.2 to 26.4%) than banana stems and leaves (8.2% crude protein, 42.8% acid detergent fiber, and 5.1% water-soluble carbohydrate). Compared with banana stem and leaf silages, the pineapple and papaya peel silages were well preserved with a lower pH and higher lactate content. The study suggests that the fruit residues contain excellent LAB species and abundant feed nutrients, and that they can be preserved as silage to be potential food resources for livestock. PMID:27108171

  7. Novel 2,7-Substituted (S)-1,2,3,4-Tetrahydroisoquinoline-3-carboxylic Acids: Peroxisome Proliferator-Activated Receptor γ Partial Agonists with Protein-Tyrosine Phosphatase 1B Inhibition.

    Otake, Kazuya; Azukizawa, Satoru; Takeda, Shigemitsu; Fukui, Masaki; Kawahara, Arisa; Kitao, Tatsuya; Shirahase, Hiroaki

    2015-01-01

    A novel series of 2,7-substituted 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid derivatives were synthesized and biologically evaluated. (S)-2-(2-Furylacryloyl)-7-[2-(2-methylindane-2-yl)-5-methyloxazol-4-yl]methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid tert-butylamine salt (13jE) was identified as a potent human peroxisome proliferator-activated receptor γ (PPARγ)-selective agonist (EC50=85 nM) and human protein-tyrosine phosphatase 1B (PTP-1B) inhibitor (IC50=1.0 µM). Compound 13jE partially activated PPARγ, but not PPARα or PPARδ, and antagonized farglitazar, a full PPARγ agonist. Cmax after the oral administration of 13jE at 10 mg/kg was 28.6 µg/mL (53 µM) in male Sprague-Dawley (SD) rats. Repeated administration of 13jE and rosiglitazone for 14 d at 10 mg/kg/d decreased plasma glucose and triglyceride levels significantly in male KK-A(y) mice. Rosiglitazone, but not 13jE, significantly increased the plasma volume and liver weight. In conclusion, 13jE showed stronger hypoglycemic and hypolipidemic effects and weaker hemodilution and hepatotoxic effects than rosiglitazone, suggesting that its safer efficacy may be due to its partial PPARγ agonism and PTP-1B inhibition. PMID:26633022

  8. Antibiotic Resistance of Probiotic Strains of Lactic Acid Bacteria Isolated from Marketed Foods and Drugs

    CHANG LIU; ZHUO-YANG ZHANG; KE DONG; JIAN-PING YUAN; XIAO-KUI GUO

    2009-01-01

    Objective To identify the antimicrobial resistance of commercial lactic acid bacteria present in microbial foods and drug additives by analyzing their isolated strains used for fermentation and probioties. Methods Antimicrobial susceptibility of 41 screened isolates was tested with disc diffusion and E-test methods after species-level identification. Resistant strains were selected and examined for the presence of resistance genes by PCR. Results Distribution of resistance was found in different species. All isolates were susceptible to chloramphenicol, tetracycline, ampicillin, amoxicillin/clavulanic acid, cephalothin, and imipenem. In addition, isolates resistant to vancomycin, rifampicin, streptomycin, bacitracin, and erythromycin were detected, although the incidence of resistance to these antibiotics was relatively low. In contrast, most strains were resistant to ciprofloxacin, amikacin, trimethoprim/sulphamethoxazole, and gentamycin. The genes msrC, vanX, and dfrA were detected in strains of Enterococcus faecium, Lactobacillus plantarum, Streptococcus thermophilus, and Lactococcus lactis. Conclusion Antibiotic resistance is present in different species of probiotic strains, which poses a threat to food safety. Evaluation of the safety of lactic acid bacteria for human consumption should be guided by established criteria, guidelines and regulations.

  9. Isolation and partial characterization of the gene for goose fatty acid synthase.

    Kameda, K; Goodridge, A G

    1991-01-01

    Fatty acid synthase is regulated by diet and hormones, with regulation being primarily transcriptional. In chick embryo hepatocytes in culture, triiodothyronine stimulates accumulation of enzyme and transcription of the gene. Since the 5'-flanking region of this gene is likely involved in hormonal regulation of its expression, we have isolated and partially characterized an avian fatty acid synthase gene. A genomic DNA library was constructed in a cosmid vector and screened with cDNA clones that contained sequence complementary to the 3' end of goose fatty acid synthase mRNA. A genomic clone (approximately 35 kilobase pairs (kb] was isolated, and a 6.5-kb EcoRI fragment thereof contained DNA complementary to the 3' noncoding region of fatty acid synthase mRNA. Additional cosmid libraries were screened with 5' fragments of previously isolated genomic clones, resulting in the isolation of five overlapping cosmid DNAs. The entire region of cloned DNA spans approximately 105 kb. Exon-containing fragments were identified by hybridization with end-labeled poly(A)+ RNA and by hybridization of labeled exon-containing genomic DNA fragments to fatty acid synthase mRNA. A new set of cDNA clones spanning approximately 3.2 kb was isolated from a lambda-ZAP goose liver cDNA library using the 5'-most exon-containing fragment of the 5'-most genomic DNA clone. This region of mRNA contains a 5'-untranslated sequence and a continuous open reading frame which includes a region that codes for the essential cysteine of the beta-ketoacyl synthase domain. The entire fatty acid synthase gene spans about 50 kb. The 5' 15 kb of the gene contain 7 exons. S1 nuclease and primer extension analyses were used to identify a single site for initiation of transcription, 174 nucleotides upstream from the putative translation initiation codon. Putative "TATA" and "CCAAT" boxes are located 28 and 60 base pairs (bp), respectively, upstream of the site of initiation of transcription. The 5'-flanking 597

  10. Differentiation of isolated native of desulphurizators Pseudomonas by means of the study of the profile of fatty acids

    The content of cellular fatty acids was determined by HRGC of twelve Colombian isolated Pseudomonas aeruginosa 17,18, 19, 20, 21, 22 and 103 pseudomonas sp 23,24,25,26 and 27 with desulphurization capacity, pseudomonas aeruginosa ATCC 9027 and 10145, pseudomonas sp ATCC 39327 and pseudomonas fluorescens. Fifty-three different types of fatty acids were found, among saturated and unsaturated of lineal chain, and mainly hydroxy acids and ramified. Of these, 17 have not been described in the literature for this genus. A group of 6 acids was presented with more frequency (15:0; 16:0; 16:1; 17:1; 3-OH 16:0 and 2-OH 15:0) in more than 75% of the pseudomonas studied. The studied microorganisms are related because they share the presence of some characteristic fatty acids, that which allows assuring that they belong to same taxonomic unit. The analysis cluster developed by means of plotting in dendrograms of the qualitative and quantitative contents of the acids fatty totals showed the formation of two attaches, the I conformed by ATCC 39327 and the isolated 17 and 25, and the II by Ps. Fluorescens and the isolated one 27. The dendrogram of the hydroxy acids shows the formation of four attaches, the I attache, (isolated 17 and 20), the II A (isolated 22 and 103), the II B (isolated 27 and ps. fluorescens) and the attache III (isolated 18 and 19). In that of the ramified fatty acids the formation of a main attache is observed conformed by four sub-groups, the IA (isolated 17), the I B (isolated 18 and 24), the I C (isolated 19 and 25 and Ps. fluorescens) and I D (isolated 27). These results show that the isolated 27,25,24, 19, 18 and 17, in their order, have narrow relationship to Ps. fluorescens

  11. Leishmanial phosphatase hydrolyzes phosphoproteins and inositol phosphates

    An extensively purified preparation of the predominant, tartrate-resistant acid phosphatase (ACP) from the external surface of Leishmania donovani promastigotes form catalyzes the dephosphorylation of several phosphoproteins; these include: pyruvate kinase, phosphorylase kinase and histones. However, the protein phosphatase activity of ACP is very low compared with that of other protein phosphates known to be involved in regulating various metabolic pathways. 32P-labelled inositoltriphosphate (IP3), a well-established second messenger derived from phosphatidylinositol-4,5-diphosphate (PIP2), was a substrate for the leishmanial acid phosphatase; incubation of the IP3 preparation with 13.2 milliunits (1 unit equals 1 μmol 4-methylumbelliferyl phosphate (MUP) cleaved per min at pH 5.5) of ACP at pH 5.5 for 4 hr resulted in hydrolysis of 75% of the radiolabelled substrate resulting in a mixture of inositoldiphosphate and inositolmonophosphate. In addition PIP2 was hydrolyzed rapidly by ACP at pH 5.5 (V/sub max/, 71 units/mg protein; k/sub m/, 4.16 μM). In contrast, to MUP which is hydrolzyed most rapidly at pH 5.5, PIP2 hydrolysis was optimal at pH 6.8. These observations raise the possibility that ACP could play a role in the host-phagocyte interaction by degrading the precursor of the second messenger, PIP2 or the second messenger itself, IP3

  12. Isolation and Selection of Anti-Candida albicans Metabolites Producing Lactic Acid Bacteria from Various Sources

    Tanes SUNGSRI

    2015-02-01

    Full Text Available Five hundred and fifty-two of lactic acid bacteria (LAB have been isolated and screened from fermented foods, natural sources and dairy effluents on De Mann Rogosa Sharpe (MRS agar. Fifty-one isolates, in the percentile of 9.24, produced the secondary metabolites that could inhibit the growth of Candida albicans BCC6120 by using dual culture overlay assay. The culture broth of LAB, moreover, showed anti-C. albicans activity in acidic condition at pH range of 3.0-5.0 by using agar well diffusion method. Interestingly, the isolate L-47-2 showed much more colonization surrounding the surface of sterile toothpick and test tube when growing in MRS broth. The identification of isolate L-47-2 by morphological and biochemical characteristics using API 50 CHL Test Kit and further confirmed by 16S rRNA gene sequence analysis revealed that isolate L47-2 was similar to Lactobacillus paracasei with 99% nucleotide identity.    

  13. Isolation of a new quinic acid derivative and its antibacterial modulating activity

    A.R. Gohari

    2010-03-01

    Full Text Available "nBackground and the purpose of the study: The species Hymenocrater calycinus, belongs to the plant family Lamiaceae and grows wildly in the north-east of Iran. Previously, the antimicrobial activity of the plant extracts was reported. In the present study, the bioactivity-guided fractionation of the methanol extract of H. calycinus and the combination effects of the isolated compound with cell wall active agents against S. aureus and E. coli was investigated. "n "nMethods: Column and thin layer chromatographic methods were used for isolation and purification and spectroscopic data (MS, 1H- and 13C-NMR, HMQC, HMBC and 1H-1H COSY were employed for identification of the compound isolated from the extract. A disk diffusion method was used to determine the antibacterial activity of the isolated compound against S. aureus and E. coli in comparison with 7 different antibiotics.Results: The isolated compound 1 was identified as 3-(3, 4- dihydroxyphenyl lactic acid 2-O-quinic acid. Compound 1 (500 µg/disc enhanced antibacterial effect of ampicillin, ciprofloxacin, vancomycin and cefepime against S. aureus and activated the effects of ampicillin and vancomycin against E. coli. "nConclusion: Results showed that the compound 1 was not active against both tested strains at any concentration below 1 mg/disk, and as a result the enhancing effect of the compound could be due its association with antibiotics.

  14. Isolation of Lactic Acid Bacteria Showing Antioxidative and Probiotic Activities from Kimchi and Infant Feces.

    Ji, Keunho; Jang, Na Young; Kim, Young Tae

    2015-09-01

    The purpose of this study was to investigate lactic acid bacteria with antioxidative and probiotic activities isolated from Korean healthy infant feces and kimchi. Isolates A1, A2, S1, S2, and S3 were assigned to Lactobacillus sp. and isolates A3, A4, E1, E2, E3, and E4 were assigned to Leuconostoc sp. on the basis of their physiological properties and 16S ribosomal DNA sequence analysis. Most strains were confirmed as safe bioresources through nonhemolytic activities and non-production of harmful enzymes such as β-glucosidase, β- glucuronidase and tryptophanase. The 11 isolates showed different resistance to acid and bile acids. In addition, they exhibited antibacterial activity against foodborne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Escherichia coli. Furthermore, all strains showed significantly high levels of hydrophobicity. The antioxidant effects of culture filtrates of the 11 strains included 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity, 2.2'- azino-bis (2-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity, and superoxide dismutase activity. The results revealed that most of the culture filtrates have effective scavenging activity for DPPH and ABTS radicals. All strains appeared to have effective superoxide dismutase activity. In conclusion, the isolated strains A1, A3, S1, and S3 have significant probiotic activities applicable to the development of functional foods and health-related products. These strains might also contribute to preventing and controlling several diseases associated with oxidative stress, when used as probiotics. PMID:25951843

  15. Evaluation of Mosquito Repellent Activity of Isolated Oleic Acid, Eicosyl Ester from Thalictrum javanicum.

    Gurunathan, Abinaya; Senguttuvan, Jamuna; Paulsamy, S

    2016-01-01

    To evaluate the traditional use, the mosquito repellent property of Thalictrum javanicum and to confirm the predicted larvicidal activity of the isolated compound, oleic acid, eicosyl ester from its aerial parts by PASS software, the present study was carried out using 4th instar stage larvae of the mosquitoes, Aedes aegypti (dengue vector) and Culex quinquefasciatus (filarial vector). Insecticidal susceptibility tests were conducted and the mortality rate was observed after 24 h exposure. The chitinase activity of isolated compound was assessed by using purified β-N-acetyl glucosaminidase (chitinase). Ecdysone 20-monooxygenase assay (radioimmuno assay) was made using the same larval stage of A. aegyptiand C. quinquefasciatus. The results were compared with the crude methanol extract of the whole plant. The isolated compound, oleic acid, eicosyl ester was found to be the most effective larvicide against A. aegypti (LC50/24 h -8.51 ppm) and C. quinquefasciatus (LC50/24 h - 12.5 ppm) than the crude methanol extract (LC50/24 h - 257.03 ppm and LC50/24 h - 281.83 ppm, respectively). The impact of oleic acid, eicosyl ester on reducing the activity of chitinase and ecdysone 20-monooxygenase was most prominent in both the target species, A. aegyptiand C. quinquefasciatus than the control. The results therefore suggest that the compound, oleic acid, eicosyl ester from Thalictrum javanicum may be considered as a potent source of mosquito larvicidal property. PMID:27168688

  16. ISOLATION AND IDENTIFICATION OF LACTIC ACID PRODUCING BACTERIA FROM CAMEL MILK

    Toqeer Ahmad, Rashida Kanwal, Izhar Hussain Athar1, Najam Ayub

    2002-03-01

    Full Text Available Lactic acid bacteria (LAB were isolated from camel milk by culturing the camel milk on specific media and pure culture was obtained by sub culturing. Purification of culture was confirmed by Gram's staining and identified by different bio-chemical tests. Camel milk contains lactic acid producing bacteria including Strpptococci such as S. cremoris and S. lactis and Lactobacilli such as L. acidophilus L. acidophilus grows more rapidly in camel milk than others as its growth is supported by camel milk. A variety of food can be preserved by lactic acid fermentation, so starter culture was prepared from strains which were isolated from camel milk. Camel and buffalo's milk cheese was prepared by using starter culture. The strains isolated from camel milk were best for acid production and can coagulate the milk in less lime. Camel milk cheese was prepared and compared with buffalo's milk cheese. It is concluded that cheese can be prepared successfully from camel milk and better results can be obtained by coagulating milk with starter culture.

  17. Evaluation of mosquito repellent activity of isolated oleic acid, eicosyl ester from Thalictrum javanicum

    Abinaya Gurunathan

    2016-01-01

    Full Text Available To evaluate the traditional use, the mosquito repellent property of Thalictrum javanicumand to confirm the predicted larvicidal activity of the isolated compound, oleic acid, eicosyl ester from its aerial parts by PASS software, the present study was carried out using 4th instar stage larvae of the mosquitoes, Aedes aegypti(dengue vector and Culex quinquefasciatus(filarial vector. Insecticidal susceptibility tests were conducted and the mortality rate was observed after 24 h exposure. The chitinase activity of isolated compound was assessed by using purified β-N-acetyl glucosaminidase (chitinase. Ecdysone 20-monooxygenase assay (radioimmuno assay was made using the same larval stage of A. aegyptiand C. quinquefasciatus. The results were compared with the crude methanol extract of the whole plant. The isolated compound, oleic acid, eicosyl ester was found to be the most effective larvicide against A. aegypti (LC50/24 h -8.51 ppm and C. quinquefasciatus (LC50/24 h - 12.5 ppm than the crude methanol extract (LC50/24 h - 257.03 ppm and LC50/24 h - 281.83 ppm, respectively. The impact of oleic acid, eicosyl ester on reducing the activity of chitinase and ecdysone 20-monooxygenase was most prominent in both the target species, A. aegyptiand C. quinquefasciatusthan the control. The results therefore suggest that the compound, oleic acid, eicosyl ester from Thalictrum javanicummay be considered as a potent source of mosquito larvicidal property.

  18. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    Khalid M. Abu-Salah

    2013-05-01

    Full Text Available The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS we screened for and isolated several microalgal strains from samples collected from the Red Sea. Relying on the fluorescence of BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diazasindacene and growth performance, four promising candidates were identified and the total lipid content and fatty acid profile was assessed for biofuels production. Selected isolates were classified as chlorophytes, belonging to three different genera: Picochlorum, Nannochloris and Desmochloris. The lipid contents were assessed microscopically by means of BODIPY 505/515-associated fluorescence to detect intracellular lipid bodies, which revealed several lipid drops in all selected strains. This result was confirmed by lipid gravimetric determination, which demonstrated that all strains under study presented inner cell lipid contents ranging from 20% to 25% of the biomass dry weight. Furthermore, the fatty acid methyl esters profile of all strains seems ideal for biodiesel production due to a low degree of polyunsaturated fatty acid methyl esters and high amount of palmitic and oleic acids.

  19. Antimicrobial Property of 2-Hydroxypropane-1,2,3-Tricarboxylic Acid Isolated from Citrus microcarpa Extract

    Seong Wei Lee; Musa Najiah

    2009-01-01

    This article described antimicrobial property and structure analysis of 2-hydroxypropane-1,2,3-tricarboxylic acid isolated from the crude extract of Citrus microcarpa. Presently, there was no report on compound from C. microcarpa that possessed antimicrobial property against fish pathogenic bacteria. Therefore, in this study, the bioactive principle in C. microcarpa extract was isolated using thin layer chromatography. It's structure was elucidated based on nuclear magnetic resonance (NMR) spectroscopic data, such as proton NMR (1HNMR), correlation spectroscopy, carbon 13 NMR, and heteronuclear multiple bond correlation data. This study showed that the bioactive compound isolated from C. microcarpa was 2-hydroxypropane-1,2,3-tricarboxylic acid monohydrate. The minimum inhibitory concentration (MIC) values of crude C. microcarpa extract and its bioactive component, 2-hydroxypropane-1,2,3-tricarboxylic acid as well as commercially available synthetic 2-hydroxypropane-1,2,3-tricarboxylic acid, were determined against 18 isolates of Edwardsiella tarda and 7 bacterial reference strains, namely, Escherichia coli (ATCC 25922), Citrobacterfreundii (ATCC 8090), Aeromonas hydrophila (ATCC 49140), Pseudomonas aeruginosa (ATCC 35032), Streptococcus agalatiae (ATCC 13813), E. tarda (ATCC 15947), and Yersinia enterocolitica (ATCC 23715), using two-fold microdilution method. The MIC values for both the natural 2-hydroxypropane-1,2,3-tricarboxylic acid and the synthetic one were ranging from 15.6 to 62.5 mg mL-1, whereas that of the crude extract was ranging from 7.8 to 31.3 mg mL-1. These findings showed that both the crude extract and its bioactive component might have potential as antimicrobial agent for aquaculture use.

  20. Identification of lactic acid bacteria isolated from Tarhana, a traditional Turkish fermented food

    Sengun, Ilkin Yucel; Nielsen, Dennis Sandris; Karapinar, Mehmet;

    2009-01-01

    Tarhana is a traditional fermented product produced from a mixture of spontaneously fermented yogurt and wheat flour in Turkey. The aims of the present study were to enumerate and identify for the first time by molecular biology-based methods predominant lactic acid bacteria (LAB) isolated during...... processing of Tarhana. Samples were collected from eight different regions of Turkey. In order to explore the relationship between raw material and the microbiology of Tarhana, yogurt and wheat flour were also analyzed. A total of 226 Gram-positive and catalase-negative isolates were obtained from MRS, M17...... and S. thermophilus was found to be the yogurt....

  1. Diversity of lactic acid bacteria isolated from raw milk in Elsharkia province, Egypt

    Alnakip, Mohamed E. A.; Mohamed, Asmaa S.; Kamal, Rania M.; Elbadry, Seham

    2016-01-01

    A total of 50 raw cow’s milk samples were collected from different areas of Elsharkia province, Egypt for characterizing lactic acid bacteria (LAB) load. Using 16S rRNA gene sequencing, a total of 41 LAB isolates have been identified corresponding to Enterococcus sp. (51.22 %) as the most predominant LAB genus, followed in order by Aerococcus (26.82 %), Lactococcus (7.32 %), Lactobacillus (7.32 %), Leuconostic (4.88 %) and Pediococcus (2.44 %) genera. All isolates were identified to species l...

  2. Characterization, Identification and Application of Lactic Acid Bacteria Isolated from Forage Paddy Rice Silage

    Ni, Kuikui; Wang, Yanping; Li, Dongxia; Cai, Yimin; Pang, Huili

    2015-01-01

    There has been growing interest to develop forage rice as a new feed resource for livestock. This study was to characterize the natural population of lactic acid bacteria (LAB) and select potentially excellent strains for paddy rice silage preparation in China. One hundred and twenty-six strains were isolated and screened from paddy rice silage prepared using a small-scale fermentation system, and ninety-nine of these isolates were considered to be LAB based on their Gram-positive and catalas...

  3. Lactic Acid Bacterial Starter Culture with Antioxidant and γ-Aminobutyric Acid Biosynthetic Activities Isolated from Flatfish-Sikhae Fermentation.

    Won, Yeong Geol; Yu, Hyun-Hee; Chang, Young-Hyo; Hwang, Han-Joon

    2015-12-01

    The aim of this study is to select a lactic acid bacterial strain as a starter culture for flatfish-Sikhae fermentation and to evaluate its suitability for application in a food system. Four strains of lactic acid bacteria isolated from commercial flatfish-Sikhae were identified and selected as starter culture candidates through investigation of growth rates, salt tolerance, food safety, and functional properties such as antioxidative and antimicrobial activities. The fermentation properties of the starter candidates were also examined in food systems prepared with these strains (candidate batch) in comparison with a spontaneous fermentation process without starter culture (control batch) at 15°C. The results showed that the candidate YG331 batch had better fermentation properties such as viable cell count, pH, and acidity than the other experimental batches, including the control batch. The results are expressed according to selection criteria based on a preliminary sensory evaluation and physiochemical investigation. Also, only a small amount of histamine was detected with the candidate YG331 batch. The radical scavenging activity of the candidate batches was better compared with the control batch, and especially candidate YG331 batch showed the best radical scavenging activity. Also, we isolated another starter candidate (identified as Lactobacillus brevis PM03) with γ-aminobutyric acid (GABA)-producing activity from commercial flatfish-Sikhae products. The sensory scores of the candidate YG331 batch were better than those of the other experimental batches in terms of flavor, color, and overall acceptance. In this study, we established selection criteria for the lactic acid bacterial starter for the flatfish-Sikhae production and finally selected candidate YG331 as the most suitable starter. PMID:26348620

  4. Efficacy of Locally Isolated Lactic Acid Bacteria Against Antibiotic-Resistant Uropathogens

    Manzoor, Asma; Ul-Haq, Ikram; Baig, Shahjhan; Qazi, Javed Iqbal; Seratlic, Sanja

    2016-01-01

    Background: Antibiotic resistance represents a serious global health threat to public health, so infections such as pneumonia and urinary tract infection (UTI) are becoming harder to treat. Therefore, it is necessary to develop an action plan to restrain the problem of antibiotic resistance. One approach in UTI control could be the use of lactobacilli because these indigenous inhabitants in human intestine have been found to play an important role in protecting the host from various infections. Objectives: We sought to check the efficacy of locally isolated Lactobacillus species to eradicate antibiotic-resistant pathogenic bacteria causing UTI. Materials and Methods: Lactic acid bacteria isolated from spoiled fruits and vegetables and grown in MRS medium were screened against multi-drug-resistant Candida albicans, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Enterococcus fecalis. Results: Fifty-four lactic acid bacteria were isolated from spoiled fruits and vegetables, of which 11 Gram-positive and catalase-negative Lactobacillus isolates were identified by carbohydrate assimilation profiles as Lactobacillus acidophilus, L. paracasei, L. delbrueckii, L. casei, L. helveticus, L. brevis, L. salivarius, L. fermentum, L. rhamnosus, L. animalis, and L. plantarum. The latter organism had the highest abundance of all the samples, so its isolates were also verified through 16S rRNA gene sequencing. The isolated Lactobacilli were screened against multi-drug-resistant uropathogens, viz. C. albicans, P. aeruginosa, K. pneumoniae, E. fecalis, and E. coli. The growth inhibition zone (GIZ) was over 10 mm against all the uropathogenic test organisms, where L. fermentum and L. plantarum strains demonstrated remarkable inhibitory activities against E. coli and E. faecalis, with a GIZ up to 28 mm. The susceptibility test to 16 antibiotics showed multidrug resistance (3 to 5 antibiotics) among all the tested uropathogens. Conclusions: The obtained results

  5. In vitro differential activity of phospholipases and acid proteinases of clinical isolates of Candida

    Aurean D'Eça Júnior

    2011-06-01

    Full Text Available INTRODUCTION: Candida yeasts are commensals; however, if the balance of normal flora is disrupted or the immune defenses are compromised, Candida species can cause disease manifestations. Several attributes contribute to the virulence and pathogenicity of Candida, including the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate the in vitro activity of phospholipases and acid proteinases in clinical isolates of Candida spp. METHODS: Eighty-two isolates from hospitalized patients collected from various sites of origin were analyzed. Phospholipase production was performed in egg yolk medium and the production of proteinase was verified in a medium containing bovine serum albumin. The study was performed in triplicate. RESULTS: Fifty-six (68.3% of isolates tested were phospholipase positive and 16 (44.4% were positive for proteinase activity. C. tropicalis was the species with the highest number of positive isolates for phospholipase (91.7%. Statistically significant differences were observed in relation to production of phospholipases among species (p<0,0001 and among the strains from different sites of origin (p=0.014. Regarding the production of acid protease, the isolates of C. parapsilosis tested presented a larger number of producers (69.2%. Among the species analyzed, the percentage of protease producing isolates did not differ statistically (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901. CONCLUSIONS: The majority of C. non-albicans and all C. albicans isolates were great producers of hydrolytic enzymes and, consequently, might be able to cause infection under favorable conditions.

  6. Antibacterial activity of mupirocin (pseudomonic Acid A) against, clinical isolates of methicillin-resistant staphylococcus aureus

    Colonized patients and health care workers are the main source of spread of methicillin resistant Staphylococcus aureus (MRSA) in hospitals. The elimination of nasal colonized MRSA plays a crucial role in infection control protocols. Mupirocin (pseudomonic acid A) is used for eradication of MRSA nasal carriage. Increasing use of pseudomonic acid A (mupirocin) has led to emergence of resistance. Objective To determine low and high level resistance of MRSA isolates from clinical specimens against mupirocin. Place and duration of study: Study was conducted at Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi from July 2006 to June 2007. Material and methods Three hundred methicillin-resistant Staphylococcus aureus isolates were studied. All clinical specimens were processed for culture and sensitivity. Staphylococcus aureus isolates were tested for methicillin resistance using 1 micro g oxacillin disk. The isolates were further tested by PCR for the presence of mecA gene. Minimum Inhibitory Concentration (MIC) of mupirocin against MRSA isolates was determined using agar dilution technique. Results Out of 300 MRSA isolates, 98% were found to have MlC against mupirocin as smaller than 4 micro g/mL. Remaining 2% isolates revealed low level resistance (MIC greater than 8 micro g/mL to 256 micro g/mL), no high level resistance (MIC greater than 512 micro g/mL) against mupirocin was detected. Conclusions: High level mupirocin resistance has not emerged so far in our setup. Due to increasing use of mupirocin, emergence of resistance against mupirocin among MRSA is a strong possibility. Strategy encompassing rational use of antimicrobials, hospital infection control, surveillance for the detection of mupirocin resistance and judicious use of this agent is required. (author)

  7. Production of lactic acid using a new homofermentative Enterococcus faecalis isolate.

    Subramanian, Mohan Raj; Talluri, Suvarna; Christopher, Lew P

    2015-03-01

    Lactic acid is an intermediate-volume specialty chemical for a wide range of food and industrial applications such as pharmaceuticals, cosmetics and chemical syntheses. Although lactic acid production has been well documented, improved production parameters that lead to reduced production costs are always of interest in industrial developments. In this study, we describe the production of lactic acid at high concentration, yield and volumetric productivity utilizing a novel homofermentative, facultative anaerobe Enterococcus faecalis CBRD01. The highest concentration of 182 g lactic acid l(-1) was achieved after 38 h of fed-batch fermentation on glucose. The bacterial isolate utilized only 2-13% of carbon for its growth and energy metabolism, while 87-98% of carbon was converted to lactic acid at an overall volumetric productivity of 5 g l(-1)  h(-1). At 13 h of fermentation, the volumetric productivity of lactate production reached 10.3 g l(-1)  h(-1), which is the highest ever reported for microbial production of lactic acid. The lactic acid produced was of high purity as formation of other metabolites was less than 0.1%. The present investigation demonstrates a new opportunity for enhanced production of lactic acid with potential for reduced purification costs. PMID:24894833

  8. Radiation-induced increase in the release of amino acids by isolated, perfused skeletal muscle

    Local exposure of the hindquarter of the rat to 15Gy of gamma-radiation resulted, 4-6h after irradiation, in increased release of amino acids by the isolated, perfused hindquarter preparation, 70% of which is skeletal muscle. This increase in release involves not only alanine and glutamine, but also those amino acids not metabolized by muscle and, therefore, released in proportion to their occurrence in muscle proteins. Because metabolic parameters and content of energy-rich phosphate compounds in muscle remain unchanged, it is unlikely that general cellular damage is the underlying cause of the radiation-induced increase in amino acid release. The findings strongly favour the hypothesis that increased availability of amino acids results from enhanced protein break-down in skeletal muscle which has its onset shortly after irradiation. This radiation-induced disturbance in protein metabolism might be one of the pathogenetic factors in the aetiology of radiation myopathy. (author)

  9. Molecular characteristics of humic acids isolated from vermicomposts and their relationship to bioactivity.

    Martinez-Balmori, Dariellys; Spaccini, Riccardo; Aguiar, Natália Oliveira; Novotny, Etelvino Henrique; Olivares, Fábio Lopes; Canellas, Luciano Pasqualoto

    2014-11-26

    Vermitechnology is an effective composting method, which transforms biomass into nutrient-rich organic fertilizer. Mature vermicompost is a renewable organic product containing humic substances with high biological activity. The aim of this study was to assess the chemical characteristics and the bioactivity of humic acids isolated from different vermicomposts produced with either cattle manure, sugar cane bagasse, sunflower cake from seed oil extraction, or filter cake from a sugar cane factory. More than 200 different molecules were found, and it was possible to identify chemical markers on humic acids according to the nature of the organic source. The large hydrophobic character of humic extracts and the preservation of altered lignin derivatives confer to humic acids the ability to induce lateral root emergence in maize seedlings. Humic acid-like substances extracted from plant biomass residues represent an additional valuable product of vermicomposting that can be used as a plant growth promoter. PMID:25379603

  10. Protein phosphatase 2A associates with Rb2/p130 and mediates retinoic acid-induced growth suppression of ovarian carcinoma cells

    Vuocolo, Scott; Purev, Enkhtsetseg; Zhang, Dongmei;

    2003-01-01

    Levels of Rb2/p130 protein are increased 5-10-fold following all-trans-retinoic acid (ATRA) treatment of the retinoid-sensitive ovarian adenocarcinoma cell line CAOV3, but not the retinoid-resistant adenocarcinoma cell line SKOV3. We found that this increase in Rb2/p130 protein levels in ATRA...

  11. Reducing Ammonia Loss from Urea by Mixing with Humic and Fulvic Acids Isolated from Coal

    Ameera A. Reeza

    2009-01-01

    Full Text Available Problem statement: Ammonia volatilization is a major pathway for nitrogen loss from surface applied urea. While all top-dressed ammonia and ammonium based N fertilizers can volatilize, the potential loss is greatest with urea and fluids containing urea. As much as 20-50% of N applied to soils is lost through volatilization alone. Thus, the objective of this laboratory study was to reduce ammonia loss from urea via mixing with humic and fulvic acids isolated from coal. Approach: This study compared four different types of treatments which were urea without additives (T1, urea with humic acid-powdered form (T2, urea with fulvic acid-liquid form (T3 and urea with humic and fulvic acids-liquid form (T4. Comparisons were made based on ammonia loss, soil NH4 and NO3- contents as well as exchangeable cations in the treated soils. Soil samples from typic paleudults (Bekenu series were used. Humic substances were isolated using standard procedures. Daily ammonia loss from soil was measured using a modified closed-dynamic air flow system method. Results: All of the treatments with humic substances significantly reduced ammonia loss ranging between 13 and 25% compared to urea alone. The treatment with both humic and fulvic acids (T4 showed pronounced ammonia loss reduction. All treatments with humic substances significantly increased NH4+ and NO3- content in soil samples compared to urea alone except for treatment having humic acid alone (T2. Treatments with fulvic acid (T3 and T4 also showed significant increase in exchangeable K+ and Na+ compared to urea alone. The increase in the formation of NH4+ over NH3, soil exchangeable cations and temporary reduction of soil pH may had retarded urea hydrolysis in the immediate vicinity of the fertilizer. Conclusion: Surface applied urea fertilizer efficiency could be increased if applied together with humic and fulvic acids.

  12. ANTIBIOTIC RESISTANCE IN LACTIC ACID BACTERIA ISOLATED FROM FERMENTED DAIRY PRODUCTS AND BOZA

    Gamze Başbülbül

    2015-06-01

    Full Text Available In this study, the resistance of 83 strains of lactic acid bacteria isolated from Turkish cheese, yogurt, kefir and boza samples to 6 antibiotics (gentamicin, tetracycline, chloramphenicol, erythromycin, vancomycin and ciprofloxacin was evaluated. The 83 isolates were identified by 16S rRNA gene sequencing and according to BLAST comparisons with sequences in the data banks, those strains showing the highest similarities with the isolates were Enterococcus faecium (10, Lactococcus lactis subsp. Lactis (10, Lactobacillus fermentum (6, Lactobacillus plantarum (6, Lactobacillus coryniformis (7, Lactobacillus casei (13, Leuconostoc mesenteroides (14, Pediococcus pentosaceus (10, Weisella confusa (7. Antimicrobial resistance of strains to 6 antibiotics was determined using the agar dilution method. The antibiotic resistance among all the isolates was detected against chloramphenicol (31,3 % of the isolates, tetracycline (30,1 %, erythromycin (2,4 %, ciprofloxacin (2,41%, vancomycin (73,5 %, intrinsic resistance. Overall 19,3 % of the isolates showed resistance against multiple antibiotics. Antibiotic resistance genes were studied by PCR and the following genes were detected; tet(M gene in Lactobacillus fermentum (1, Lactobacillus plantarum (1, Pediococcus pentosaceus (5, Enterococcus faecium (2, Weisella confusa (4 and the vancomycin resistance gene van(A in one Weisella confusa strain.

  13. A Fatty Acid Glycoside from a Marine-Derived Fungus Isolated from Mangrove Plant Scyphiphora hydrophyllacea

    Wen-Li Mei

    2012-03-01

    Full Text Available To study the antimicrobial components from the endophytic fungus A1 of mangrove plant Scyphiphora hydrophyllacea Gaertn. F., a new fatty acid glucoside was isolated by column chromatography from the broth of A1, and its structure was identified as R-3-hydroxyundecanoic acid methylester-3-O-α-l-rhamnopyranoside (1 by spectroscopic methods including 1D and 2D NMR (HMQC, 1H-1H COSY and HMBC and chemical methods. Antimicrobial assay showed compound 1 possessed modest inhibitory effect on Saphylococcus aureus and methicillin-resistant S. aureus (MRSA using the filter paper disc agar diffusion method.

  14. Reduction of Ammonia Loss from Urea through Mixing with Humic Acids Isolated from Peat Soil (Saprists)

    Regis Bernard; Osumanu H. Ahmed; Nik M.A. Majid; Mohamadu B. Jalloh

    2009-01-01

    Problem statement: Application of urea as a source of nitrogen fertilizer has an adverse effect on ammoniacal loss to the environment. This study was conducted to reduce ammonia loss from urea by mixing with Humic Acids (HA) isolated from Saprists peat. Approach: The effects of urea amended with four different amounts of humic acids, 0.25, 0.50, 0.75 and 1.00 g were evaluated in laboratory conditions using a closed dynamic air flow system. The mineral soil that was used as medium for the stud...

  15. Nucleotide and amino acid sequences of a coat protein of an Ukrainian isolate of Potato virus Y: comparison with homologous sequences of other isolates and phylogenetic analysis

    Budzanivska I. G.

    2014-03-01

    Full Text Available Aim. Identification of the widespread Ukrainian isolate(s of PVY (Potato virus Y in different potato cultivars and subsequent phylogenetic analysis of detected PVY isolates based on NA and AA sequences of coat protein. Methods. ELISA, RT-PCR, DNA sequencing and phylogenetic analysis. Results. PVY has been identified serologically in potato cultivars of Ukrainian selection. In this work we have optimized a method for total RNA extraction from potato samples and offered a sensitive and specific PCR-based test system of own design for diagnostics of the Ukrainian PVY isolates. Part of the CP gene of the Ukrainian PVY isolate has been sequenced and analyzed phylogenetically. It is demonstrated that the Ukrainian isolate of Potato virus Y (CP gene has a higher percentage of homology with the recombinant isolates (strains of this pathogen (approx. 98.8– 99.8 % of homology for both nucleotide and translated amino acid sequences of the CP gene. The Ukrainian isolate of PVY is positioned in the separate cluster together with the isolates found in Syria, Japan and Iran; these isolates possibly have common origin. The Ukrainian PVY isolate is confirmed to be recombinant. Conclusions. This work underlines the need and provides the means for accurate monitoring of Potato virus Y in the agroecosystems of Ukraine. Most importantly, the phylogenetic analysis demonstrated the recombinant nature of this PVY isolate which has been attributed to the strain group O, subclade N:O.

  16. HUMIC ACID-LIKE MATTER ISOLATED FROM GREEN URBAN WASTES. PART I: STRUCTURE AND SURFACTANT PROPERTIES

    Enzo Montoneri

    2008-02-01

    Full Text Available A humic acid-like substance (cHAL2 isolated from urban green wastes before composting was compared to a humic acid-like substance (cHAL isolated from a mix of urban organic humid waste fraction and green residues composted for 15 days. cHAL2 was found to contain more aliphatic and O-alkyl C atoms relative to aromatic, phenol, and carboxyl C atoms, and to yield higher critical micellar concentration (cmc = 0.97 g L-1 and surface tension at the cmc (cmc = 37.8 mN/min water than cHAL (cmc = 0.40 g L-1; cmc = 36.1 mN/m. The results point out that biomass wastes may be an interesting source of biosurfactants with diversified properties that depend on the nature of waste and on its process of treatment.

  17. Isolation and characterization of lactic acid bacteria and yeasts from the Brazilian grape sourdough

    Krischina Singer Aplevicz

    2014-04-01

    Full Text Available Sourdough is a mixture of flour and water fermented by lactic acid bacteria and yeast, with a large use in bakery products. This study was developed with Brazilian grape (Niagara rosada sourdough obtained from spontaneous fermentation. The aim of this work was to characterize genotypic and phenotypically lactic acid bacteria and yeasts isolated from sourdough. The phenotypic identification for bacteria and yeasts was performed by using the kit API50CHL and 20CAUX and the genotypic characterization was performed by sequencing method. A total of four isolated strains were analyzed in this study. Two of these strains were phenotypically and genotypic identified as Lactobacillus paracasei and one as Saccharomyces cerevisiae. Another sample phenotypically identified as Candida pelliculosa did not show the same identity by sequencing. It shows the need to use phenotypic and genotypic characterization associated for the correct microorganism identification.

  18. Utilization of Vinegar for Isolation of Cellulose Producing Acetic Acid Bacteria

    Wastes of traditionally fermented Turkish vinegar were used in the isolation of cellulose producing acetic acid bacteria. Waste material was pre-enriched in Hestrin-Schramm medium and microorganisms were isolated by plating dilution series on HS agar plates The isolated strains were subjected to elaborate biochemical and physiological tests for identification. Test results were compared to those of reference strains Gluconacetobacter xylinus DSM 46604, Gluconacetobacter hansenii DSM 5602 and Gluconacetobacter liquefaciens DSM 5603. Seventeen strains, out of which only three were found to secrete the exopolysaccharide cellulose. The highest cellulose yield was recorded as 0.263±0.02 g cellulose L-1 for the strain AS14 which resembled Gluconacetobacter hansenii in terms of biochemical tests.

  19. Production of bacteriocin-like substances by lactic acid bacteria isolated from regional ovine cheese

    Cássia Regina Nespolo

    2010-12-01

    Full Text Available Lactic acid bacteria (LAB were isolated from ovine milk and cheeses manufactured in the South Region of Brazil. Among 112 bacterial isolates investigated, 59 were chosen through a screening for LAB. Among these 59 strains of LAB, 21% showed antimicrobial, proteolytic and lipolytic activities. Based on this screening, Lactobacillus plantarum LCN 17 and Lactobacillus rhamnosus LCN 43 were selected and tested for the production of bacteriocin-like substances (BLS. The BLS produced by both isolates showed antimicrobial activity against Listeria monocytogenes, whereas that produced by L. plantarum LCN 17 presented higher stability to different temperature, pH and enzyme treatments. These strains present potential for production of BLS, and for use as starter cultures.

  20. Surface chemical properties of sodium salts of carboxylic acids isolated from Green River shale. [Sodium carboxylates

    McKay, J.F.; Blanche, M.S.; Robertson, R.E.

    1985-12-01

    Organic material isolated from Green River shale varies substantially with the method of isolation. Short-time supercritical fluid treatment and solvent extraction of Green River shale produces large amounts of sodium carboxylates. These sodium salts were observed to form emulsions and therefore be surface active. Quantitative surface activity measurements were then determined using the shale extract. The material was found to have a limiting surface tension of about 41 dynes/cm (as expected) for carboxylates. However, the critical micelle concentration is quite high and has a measured molecular weight value of 600. This probably results from higher solubility of the lower molecular weight species. The solution did not display hysteresis. In general the carboxylic acid salts isolated from Green River shale displayed surface activity similar to those of model compounds cited in the literature.

  1. Utilization of Vinegar for Isolation of Cellulose Producing Acetic Acid Bacteria

    Aydin, Y. Andelib; Aksoy, Nuran Deveci

    2010-06-01

    Wastes of traditionally fermented Turkish vinegar were used in the isolation of cellulose producing acetic acid bacteria. Waste material was pre-enriched in Hestrin-Schramm medium and microorganisms were isolated by plating dilution series on HS agar plates The isolated strains were subjected to elaborate biochemical and physiological tests for identification. Test results were compared to those of reference strains Gluconacetobacter xylinus DSM 46604, Gluconacetobacter hansenii DSM 5602 and Gluconacetobacter liquefaciens DSM 5603. Seventeen strains, out of which only three were found to secrete the exopolysaccharide cellulose. The highest cellulose yield was recorded as 0.263±0.02 g cellulose L-1 for the strain AS14 which resembled Gluconacetobacter hansenii in terms of biochemical tests.

  2. Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows’ milk cheeses

    Terzić-Vidojević Amarela; Mihajlović Sanja; Uzelac Gordana; Golić Nataša; Fira Đ.; Kojić M.; Topisirović Lj.

    2014-01-01

    The aim of this study was to identify and characterize the lactic acid bacteria (LAB) of artisanal Golija raw and cooked cows’ milk cheeses traditionally manufactured without the addition of starter culture. A total of 188 Gram-positive and catalase-negative isolates of Golija cheeses were obtained from seven samples of different ripening time. Phenotypebased assays as well as rep-PCR and 16S rDNA sequence analysis were undertaken for all 188 Lstrains. The ...

  3. Evaluation of Mosquito Repellent Activity of Isolated Oleic Acid, Eicosyl Ester from Thalictrum javanicum

    Abinaya Gurunathan; Jamuna Senguttuvan; S Paulsamy

    2016-01-01

    To evaluate the traditional use, the mosquito repellent property of Thalictrum javanicum and to confirm the predicted larvicidal activity of the isolated compound, oleic acid, eicosyl ester from its aerial parts by PASS software, the present study was carried out using 4th instar stage larvae of the mosquitoes, Aedes aegypti (dengue vector) and Culex quinquefasciatus (filarial vector). Insecticidal susceptibility tests were conducted and the mortality rate was observed after 24 h exposure. Th...

  4. Lactic acid bacteria from Jijel's traditional butter: Isolation, identification and major technological traits

    Idoui, Tayeb; Karam, Nour-Eddine

    2008-01-01

    Twenty seven (27) lactic acid bacteria were isolated from Jijel’s traditional butter. These strains belong to three genera: Lactococcus, Lactobacillus and Leuconostoc. The results showed that Lactobacillus plantarum was the predominant species in this traditional butter. It appears that these strains have some interesting technological properties.Se aíslan veintisiete (27) bacterias acidolácticas de la mantequilla tradicional de Jijel. Éstas pertenecen a los géneros Lactococcus, Lactobacill...

  5. ISOLATION OF LACTIC ACID BACTERIA UNDER LOW TEMPERATURE FOR THE PREPARATION OF YOGURT

    Javid Ahmad Bhat

    2014-02-01

    Full Text Available An investigation of isolation of Lactic acid bacteria was carried out under low temperature for the preparation of Yogurt by using various food supplements like carrot, ground-nut and tomato juices. Methods: Various samples of Cow milk, Skimmed milk were processed along with nutrients like Carrot, ground nut and tomato juices with Tryptone glucose yeast extract agar (TGYA at different temperatures like 50C, 150C and 220C for the isolation of Lactic acid bacteria for the preparation of yogurt. The characteristic isolates were identified by using various biochemical tests and direct microscopy. Results: Lactic acid bacteria (LAB dominated the microbial population of Yogurt, and were identified according to their morphological and physiological characteristics. Among these lactobacilli were frequently occurring organisms. The most abundant species were Lactobacillus delbrueckii subspecies Bulgaricus and Streptococcus thermophilus. The Lactic Streptococci was subjected to bio-chemical tests to identify the species. Based on the biochemical reactions the species was identified as Lactococcus Lactis, sub species di-acetylactis. Isolated culture of lactic Streptococci was found to grow at low temperature. When this was used as an inoculum to prepare yogurt at 50C, 150C and 220C curdling took place in 3days time. In order to reduce the setting time, nutrients in the form of carrot, ground-nut and tomato juices were added. The yogurt was found to set at 50C in 30hrs which is considered useful. Acidity of yogurt was found to be 0.53%- 0.55%. The yogurt was found to contain di-acetyl and quality of yogurt was good.

  6. Screening of biogenic amine production by lactic acid bacteria isolated from grape musts and wine

    Moreno-Arribas, M. Victoria; Polo, María Carmen; Jorganes, Felisa; Muñoz, Rosario

    2003-01-01

    The potential to produce the biogenic amines tyramine, histamine and putrescine, was investigated for lactic acid bacteria (LAB) of various origin, including commercial malolactic starter cultures, type strains and 78 strains isolated from Spanish grape must and wine. The presence of biogenic amines in a decarboxylase synthetic broth was determined by reverse-phase high performance liquid chromatography (RP-HPLC). Tyramine was the main amine formed by the LAB strains investigated. ...

  7. Molecular characterization of lactic acid bacteria isolated from industrially fermented Greek table olives

    Doulgeraki, Agapi; Pramateftaki, Paraskevi; Argyri, Anthoula; Nychas, George John; Tassou, Chrysoula; Panagou, Efstathios

    2012-01-01

    A total of 145 lactic acid bacteria (LAB) isolates have been recovered from fermented table olives and brine and characterized at strain level with molecular tools. Pulsed-Field Gel Electrophoresis (PFGE) of ApaI macrorestriction fragments was applied for strain differentiation. Species differentiation was based either on Denaturing Gradient Gel Electrophoresis (PCR-DGGE) (black olives) or on restriction analysis of the amplified 16S rRNA gene (PCR-ARDRA) (brine and green olives). Species ide...

  8. Antifungal Activity of Phenyllactic Acid against Molds Isolated from Bakery Products

    Lavermicocca, Paola; Valerio, Francesca; Visconti, Angelo

    2003-01-01

    Phenyllactic acid (PLA) has recently been found in cultures of Lactobacillus plantarum that show antifungal activity in sourdough breads. The fungicidal activity of PLA and growth inhibition by PLA were evaluated by using a microdilution test and 23 fungal strains belonging to 14 species of Aspergillus, Penicillium, and Fusarium that were isolated from bakery products, flours, or cereals. Less than 7.5 mg of PLA ml−1 was required to obtain 90% growth inhibition for all strains, while fungicid...

  9. Detection of phosphatase activity in aquatic and terrestrial cyanobacterial strains

    Babić Olivera B.

    2013-01-01

    Full Text Available Cyanobacteria, as highly adaptable microorganisms, are characterized by an ability to survive in different environmental conditions, in which a significant role belongs to their enzymes. Phosphatases are enzymes produced by algae in relatively large quantities in response to a low orthophosphate concentration and their activity is significantly correlated with their primary production. The activity of these enzymes was investigated in 11 cyanobacterial strains in order to determine enzyme synthesis depending on taxonomic and ecological group of cyanobacteria. The study was conducted with 4 terrestrial cyanobacterial strains, which belong to Nostoc and Anabaena genera, and 7 filamentous water cyanobacteria of Nostoc, Oscillatoria, Phormidium and Microcystis genera. The obtained results showed that the activity of acid and alkaline phosphatases strongly depended on cyanobacterial strain and the environment from which the strain originated. Higher activity of alkaline phosphatases, ranging from 3.64 to 85.14 μmolpNP/s/dm3, was recorded in terrestrial strains compared to the studied water strains (1.11-5.96 μmolpNP/s/dm3. The activity of acid phosphatases was higher in most tested water strains (1.67-6.28 μmolpNP/s/dm3 compared to the activity of alkaline phosphatases (1.11-5.96 μmolpNP/s/dm3. Comparing enzyme activity of nitrogen fixing and non-nitrogen fixing cyanobacteria, it was found that most nitrogen fixing strains had a higher activity of alkaline phosphatases. The data obtained in this work indicate that activity of phosphatases is a strain specific property. The results further suggest that synthesis and activity of phosphatases depended on eco-physiological characteristics of the examined cyanobacterial strains. This can be of great importance for the further study of enzymes and mechanisms of their activity as a part of cyanobacterial survival strategy in environments with extreme conditions. [Projekat Ministarstva nauke Republike

  10. β-methyl-15-p-iodophenylpentadecanoic acid metabolism and kinetics in the isolated rat heart

    The use of 15-p-iodophenyl-β-methyl-pentadecanoic acid (βMe-IPPA) as an indicator of long chain fatty acid (LCFA) utilization in nuclear medicine studies was evaluated in the isolated, perfused, working rat heart. Time courses of radioactivity (residue curves) were obtained following bolus injections of both βMe-IPPA and its straight chain counterpart 15-p-iodophenyl-pentadecanoic acid (IPPA). IPPA kinetics clearly indicated flow independent impairment of fatty acid oxidation caused by the carnitine palmitoyltransferase I inhibitor 2[5(4-chlorophenyl)pentyl]oxirane-2-carboxylate (POCA). In contrast, βMe-IPPA kinetics were insensitive to changes in fatty acid oxidation rate and net utilization of long chain fatty acid. Analysis of radiolabeled species in coronary effluent and heart homogenates showed the methylated fatty acid to be readily incorporated into complex lipids but a poor substrate for oxidation. POCA did not significantly alter metabolism of the tracer, suggesting that the tracer is poorly metabolized beyond βMe-IPPA-CoA in the oxidative pathway. (orig.)

  11. Isolation and evaluation of antiglycation potential of polyalthic acid (furano-terpene from Daniella oliveri

    Olubunmi Atolani

    2014-12-01

    Full Text Available A furano-diterpene (polyalthic acid was isolated as a major stable compound for the first time from the oleoresin of the Daniella oliveri of the family Caesalpiniacea through column chromatography fractionation. Polyalthic acid was characterized using data obtained from EIMS, HREIMS, ESI-MS, MALDI-MS as well as 1D and 2D NMR and it was evaluated for its potential to inhibit the formation of advanced glycation end-products (AGEs using a standard in vitro antiglycation procedure. Polyalthic acid indicated a negative antiglycation potential compared to standard inhibitor that has 85% inhibition, which is an indication that polyalthic acid may not contribute to the antiglycation activity of the plant as acclaimed in folkloric medicine. The negative antiglycation observed could indicate that the polyalthic acid could trigger glycation, thereby subjecting users to various degrees of complications. The bioactivity evaluation on molinspiration evaluator indicated that polyalthic acid could be a potential drug candidate. The biological and chemical insights gained on polyalthic acid provide a good basis for future research.

  12. Ficusonic acid: a new cytotoxic triterpene isolated from Maytenus royleanus (Wall. ex M. A. Lawson) cufodontis

    Din, Ala Ud; Uddin, Ghias [Center for Phytomedicine and Medicinal Organic Chemistry, Institute of Chemical Sciences, University of Peshawar(Pakistan); Hussain, Nusrat; Choudary, Mohammad Iqbal, E-mail: allauddin77@yahoo.com [International Center for Chemical and Biological Sciences, HEJ Research Institute of Chemistry, University of Karachi (Pakistan)

    2013-04-15

    Phytochemical investigation of the roots of Maytenus royleanus resulted into the isolation of a new cytotoxic triterpene ficusonic acid, 3{beta},21{beta}-dihydroxyolean-12-en-29-oic acid, together with three known compounds, 3{alpha},22{beta}-dihydroxyolean-12-en-29-oic acid, salaspermic acid and orthosphenic acid, reported for the first time from this source. Their structures were established on the basis of extensive spectroscopic techniques. The cytotoxic activity of compound 3{beta},21{beta}-dihydroxyolean-12-en-29-oic acid was evaluated against two cancer cell lines, PC-3 prostate and HeLa cervical cancer lines. 3{beta},21{beta}-dihydroxyolean-12-en-29-oic acid showed weak activity against PC-3 (IC{sub 50} = 35.42 Greek-Small-Letter-Mu mol L{sup -1}) however against HeLa (IC{sub 50} = 20.47 Greek-Small-Letter-Mu mol L{sup -1}), its activity was moderate. (author)

  13. Protein phosphatase 1α is a Ras-activated Bad phosphatase that regulates interleukin-2 deprivation-induced apoptosis

    Ayllón, Verónica; Martínez-A, Carlos; García, Alphonse; Cayla, Xavier; Rebollo, Angelita

    2000-01-01

    Growth factor deprivation is a physiological mechanism to regulate cell death. We utilize an interleukin-2 (IL-2)-dependent murine T-cell line to identify proteins that interact with Bad upon IL-2 stimulation or deprivation. Using the yeast two-hybrid system, glutathione S-transferase (GST) fusion proteins and co-immunoprecipitation techniques, we found that Bad interacts with protein phosphatase 1α (PP1α). Serine phosphorylation of Bad is induced by IL-2 and its dephosphorylation correlates with appearance of apoptosis. IL-2 deprivation induces Bad dephosphorylation, suggesting the involvement of a serine phosphatase. A serine/threonine phosphatase activity, sensitive to the phosphatase inhibitor okadaic acid, was detected in Bad immunoprecipitates from IL-2-stimulated cells, increasing after IL-2 deprivation. This enzymatic activity also dephosphorylates in vivo 32P-labeled Bad. Treatment of cells with okadaic acid blocks Bad dephosphorylation and prevents cell death. Finally, Ras activation controls the catalytic activity of PP1α. These results strongly suggest that Bad is an in vitro and in vivo substrate for PP1α phosphatase and that IL-2 deprivation-induced apoptosis may operate by regulating Bad phosphorylation through PP1α phosphatase, whose enzymatic activity is regulated by Ras. PMID:10811615

  14. Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol

    Shu; Chen; Wang; Chen; Kai; Chang; Shu; Chang; Chan; Jiunn; Shiuh; Shieh; Chih; Kwang; Chiu; Pin-Der; Duh

    2014-01-01

    Objective:To evaluate the ability of lactic acid bacteria(LAB)strains isolated from fermented mustard to lower the cholesterol in vitro.Methods:The ability of 50 LAB strains isolated from fermented mustard on lowering cholesterol in vitro was determined by modified o-phtshalaldehyde method.The LAB isolates were analyzed for their resistance to acid and bile salt.Strains with lowering cholesterol activity,were determined adherence to Caco-2 cells.Results:Strain B0007,B0006 and B0022 assimilated more cholesterol than BCRC10474 and BCRC17010.The isolated strains showed tolerance to pH 3.0 for 3h despite variations in the degree of viability and bile-tolerant strains,with more than 10~s CFU/mL after incubation for 24 h at 1%oxigall in MRS.In addition,strain B0007 and B0022 identified as Lactobacillus plantarum with 16S rDNA sequences were able to adhere to the Caco-2 cell lines.Conclusions:These strains B0007 and B0022 may be potential functional sources for cholesterollowering activities as well as adhering to Caco-2 cell lines.

  15. Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains.

    Leite, A M O; Miguel, M A L; Peixoto, R S; Ruas-Madiedo, P; Paschoalin, V M F; Mayo, B; Delgado, S

    2015-06-01

    A total of 34 lactic acid bacteria isolates from 4 different Brazilian kefir grains were identified and characterized among a group of 150 isolates, using the ability to tolerate acidic pH and resistance to bile salts as restrictive criteria for probiotic potential. All isolates were identified by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of representative amplicons. Eighteen isolates belonged to the species Leuconostoc mesenteroides, 11 to Lactococcus lactis (of which 8 belonged to subspecies cremoris and 3 to subspecies lactis), and 5 to Lactobacillus paracasei. To exclude replicates, a molecular typing analysis was performed by combining repetitive extragenic palindromic-PCR and random amplification of polymorphic DNA techniques. Considering a threshold of 90% similarity, 32 different strains were considered. All strains showed some antagonistic activity against 4 model food pathogens. In addition, 3 Lc. lactis strains and 1 Lb. paracasei produced bacteriocin-like inhibitory substances against at least 2 indicator organisms. Moreover, 1 Lc. lactis and 2 Lb. paracasei presented good total antioxidative activity. None of these strains showed undesirable enzymatic or hemolytic activities, while proving susceptible or intrinsically resistant to a series of clinically relevant antibiotics. The Lb. paracasei strain MRS59 showed a level of adhesion to human Caco-2 epithelial cells comparable with that observed for Lactobacillus rhamnosus GG. Taken together, these properties allow the MRS59 strain to be considered a promising probiotic candidate. PMID:25841972

  16. Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol

    Shu Chen Wang; Chen Kai Chang; Shu Chang Chan; Jiunn Shiuh Shieh; Chih Kwang Chiu; Pin-Der Duh

    2014-01-01

    Objective: To evaluate the ability of lactic acid bacteria (LAB) strains isolated from fermented mustard to lower the cholesterol in vitro.Methods:The ability of 50 LAB strains isolated from fermented mustard on lowering cholesterol in vitro was determined by modified o-phtshalaldehyde method. The LAB isolates were analyzed for their resistance to acid and bile salt. Strains with lowering cholesterol activity, were determined adherence to Caco-2 cells. Results: Strain B0007, B0006 and B0022 assimilated more cholesterol than BCRC10474 and BCRC 17010. The isolated strains showed tolerance to pH 3.0 for 3 h despite variations in the degree of viability and bile-tolerant strains, with more than 108 CFU/mL after incubation for 24 h at 1%oxigall in MRS. In addition, strain B0007 and B0022 identified as Lactobacillus plantarum with 16S rDNA sequences were able to adhere to the Caco-2 cell lines.Conclusions:These strains B0007 and B0022 may be potential functional sources for cholesterol-lowering activities as well as adhering to Caco-2 cell lines.

  17. Isolation of a tannic acid-degrading Streptococcus sp. from an anaerobic shea cake digester.

    Nitiema, L W; Dianou, D; Simpore, J; Karou, S D; Savadogo, P W; Traore, A S

    2010-01-01

    An anaerobic digester fed with shea cake rich in tannins and phenolic compounds rich-shea cake and previously inoculated with anaerobic sludge from the pit of a slaughterhouse, enabled six months acclimatization of the bacteria to aromatic compounds. Afterwards, digester waste water samples were subject to successive culture on media with 1 g L(-1) tannic acid allowing the isolation of a bacterial strain coded AB. Strain AB was facultatively anaerobic, mesophilic, non-motile, non-sporulating, catalase and oxidase negative bacterium, namely strain AB, was isolated from an anaerobic digester fed with shea cake rich in tannins and phenolic compounds, after inoculation with anaerobic sludge from the pit of a slaughterhouse and enrichment on tannic acid. The coccoid cells occurred in pair, short or long chains and stained Gram-positive. Strain AB fermented a wide range of carbohydrates including glucose, fructose, galactose, raffinose, arabinose, sucrose, maltose, lactose, starch and cellulose. Optimum growth occurred with glucose and tannic acid at 37 degrees C and pH 8. The pH, temperature and salt concentration for growth ranged from 5 to 9, 20 to 45 degrees C and 0 to 15 g L(-1), respectively. Strain AB converted tannic acid to gallic acid. These features were similar to those of the Streptococcus genus. The determination of tannic acid hydrolysis end products, ability to utilize various organic acids, alcohols and peptides, GC% of the DNA, the sequencing of 16S rRNA gene and DNA-DNA hybridization will permit to confirm this affiliation and to determine the species. PMID:20415153

  18. Cloning and characterization of a novel human phosphatidic acid phosphatase type 2, PAP2d, with two different transcripts PAP2d_v1 and PAP2d_v2.

    Sun, Liyun; Gu, Shaohua; Sun, Yaqiong; Zheng, Dan; Wu, Qihan; Li, Xin; Dai, Jianfeng; Dai, Jianliang; Ji, Chaoneng; Xie, Yi; Mao, Yumin

    2005-04-01

    This study reports the cloning and characterization of a novel human phosphatidic acid phosphatase type 2 isoform cDNAs (PAP2d) from the foetal brain cDNA library. The PAP2d gene is localized on chromosome 1p21.3. It contains six exons and spans 112 kb of the genomic DNA. By large-scale cDNA sequencing we found two splice variants of PAP2d, PAP2d_v1 and PAP2d_v2. The PAP2d_v1 cDNA is 1722 bp in length and spans an open reading frame from nucleotide 56 to 1021, encoding a 321aa protein. The PAP2d_v2 cDNA is 1707 bp in length encoding a 316aa protein from nucleotide 56-1006. The PAP2d_v1 cDNA is 15 bp longer than the PAP2d_v2 cDNA in the terminal of the fifth exon and it creates different ORF. Both of the proteins contain a well-conserved PAP2 motif. The PAP2d_v1 is mainly expressed in human brain, lung, kidney, testis and colon, while PAP2d_v2 is restricted to human placenta, skeletal muscle, and kidney. The two splice variants are co-expressed only in kidney. PMID:16010976

  19. A Chronoamperometric Screen Printed Carbon Biosensor Based on Alkaline Phosphatase Inhibition for W(VI Determination in Water, Using 2-Phospho-l-Ascorbic Acid Trisodium Salt as a Substrate

    Ana Lorena Alvarado-Gámez

    2015-01-01

    Full Text Available This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3, a repeatability of 9.4% (n = 3 and a detection limit of 0.29 ± 0.01 µM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case and a dynamic range from 0.6 to 30 µM. This study was performed by means of a Lineweaver–Burk plot, showing a mixed kinetic inhibition.

  20. Tartrate-resistant acid phosphatase (TRAP) co-localizes with receptor activator of NF-KB ligand (RANKL) and osteoprotegerin (OPG) in lysosomal-associated membrane protein 1 (LAMP1)-positive vesicles in rat osteoblasts and osteocytes.

    Solberg, L B; Stang, E; Brorson, S-H; Andersson, G; Reinholt, F P

    2015-02-01

    Tartrate-resistant acid phosphatase (TRAP) is well known as an osteoclast marker; however, a recent study from our group demonstrated enhanced number of TRAP + osteocytes as well as enhanced levels of TRAP located to intracellular vesicles in osteoblasts and osteocytes in experimental osteoporosis in rats. Such vesicles were especially abundant in osteoblasts and osteocytes in cancellous bone as well as close to bone surface and intracortical remodeling sites. To further investigate TRAP in osteoblasts and osteocytes, long bones from young, growing rats were examined. Immunofluorescence confocal microscopy displayed co-localization of TRAP with receptor activator of NF-KB ligand (RANKL) and osteoprotegerin (OPG) in hypertrophic chondrocytes and diaphyseal osteocytes with Pearson's correlation coefficient ≥0.8. Transmission electron microscopy showed co-localization of TRAP and RANKL in lysosomal-associated membrane protein 1 (LAMP1) + vesicles in osteoblasts and osteocytes supporting the results obtained by confocal microscopy. Recent in vitro data have demonstrated OPG as a traffic regulator for RANKL to LAMP1 + secretory lysosomes in osteoblasts and osteocytes, which seem to serve as temporary storage compartments for RANKL. Our in situ observations indicate that TRAP is located to RANKL-/OPG-positive secretory lysosomes in osteoblasts and osteocytes, which may have implications for osteocyte regulation of osteoclastogenesis. PMID:25201349

  1. Radioimmunodetection of lymph node invasion in prostatic cancer. The use of iodine-123 (123I)-labeled monoclonal anti-prostatic acid phosphatase (PAP) 227 A F(ab')2 antibody fragments in vivo

    The therapeutic indications in prostatic cancer depend on the regional and distant extension of the cancer and are difficult to assess before lymphadenectomy. Radioimmunodetection of lymph node involvement with monoclonal anti-prostatic acid phosphatase (PAP) antibodies can be proposed as a noninvasive alternative to lymphadenectomy. Fifteen patients with various stages of histologically proven prostatic cancer were examined by immunolymphoscintigraphy (ILS) before treatment to detect lymph node metastases. These patients had Stage A (n = 7), Stage B (n = 3), Stage C (n = 2), and Stage D (n = 3) tumors. They received between 100 and 400 micrograms of monoclonal antibody 227 A in the form of F(ab')2 fragments labeled with iodine-123 (123I). The antibody was injected directly into the periprostatic area. ILS images were obtained after 1, 3, 6, and 24 hours. Three days later, each patient underwent a lymphadenectomy for histologic examination. The results of the histologic examination and ILS were compared. In ten patients, the examination did not show any images capable of being interpreted as lymphadenopathy and histologic examination confirmed the integrity of the nodes examined. In five cases, scintigraphy suggested the presence of lymph node invasion by prostatic cancer and this was confirmed by histologic examination in three of the five cases. Overall, in terms of lymphadenopathy, this examination had a sensitivity of 100% and a specificity of 83%. Therefore, ILS appears to be capable of detecting lymph node metastases in prostatic cancer

  2. Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

    Xu, Sheng Jun; Kim, Byung Sup

    2014-01-01

    In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited th...

  3. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom

    Elaine de Paula Mendonca-Franqueiro; Eliane Candiani Arantes; Marcelo Dias-Baruffi; Suely Vilela Sampaio; Raquel de Melo Alves-Paiva; Marco Aurélio Sartim; Daniel Roberto Callejon; Helder Henrique Paiva; Gilmara Ausech Antonucci; José César Rosa; Adélia Cristina Oliveira Cintra; Jo(a)o José Franco

    2011-01-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycanbinding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability,which suggested that galatrox might contribute to venom toxicity by directly inducing cell death.

  4. Characterization of lactic acid bacteria isolated from artisanal Zlatar cheeses produced at two different geographical location

    Terzić-Vidojević Amarela

    2009-01-01

    Full Text Available Eighty-one strains of lactic acid bacteria (LAB were isolated from white semi-hard homemade cheese, designated Zlatar BGNV, which was taken from household settled on Northern side of mountain Zlatar. The Zlatar BGNV cheese was manufactured from raw cow's milk without addition of the starter culture. All isolates of LAB were characterized by phenotypic and genotypic tests. Identification of strains was done by the repetitive extragenic palindromic-polymerase chain reaction (rep-PCR with (GTG5 primer and 16S rDNA sequence analysis. The most present species in Zlatar BGNV cheese were Lactobacillus casei/paracasei (65.43% and Enterococcus faecalis (29.63%. Two facultative heterofermentative rods were identified as Lactobacillus plantarum (2.47%, and two obligate hetrofermentative LAB isolates as Lactobacillus parabuchneri (2.47%. Among all 81 tested isolates, only eight enterococci were producers of antimicrobial compounds. Fourteen of 16 tested lactobacilli isolates showed medium to very good proteolytic activity. All 57 lactobacilli from the Zlatar BGNV cheese curdled milk very slowly or did not curdle milk at all. However, three isolates of enterococci, BGNV1-63, BGNV1-76 and BGNV1-80, showed very good activity in milk and curdled milk within 5 h. They showed very high proteolytic activity hydrolyzing completely αs1- and κ-casein after 3 h, and β-casein after 30 min of incubation. In addition, those three enterococcal isolates degraded gelatin. Comparing obtained results with those previously achieved in examination of LAB microflora in another Zlatar BGZLS cheese made also from raw cow's milk, it can be concluded that LAB microflora in the Zlatar BGNV cheese is less diverse.

  5. Isolation, identification and quantification of unsaturated fatty acids, amides, phenolic compounds and glycoalkaloids from potato peel.

    Wu, Zhi-Gang; Xu, Hai-Yan; Ma, Qiong; Cao, Ye; Ma, Jian-Nan; Ma, Chao-Mei

    2012-12-15

    Eleven compounds were isolated from potato peels and identified. Their structures were determined by interpretation of UV, MS, 1D, and 2D NMR spectral data and by comparison with reported data. The main components of the potato peels were found to be chlorogenic acid and other phenolic compounds, accompanied by 2 glycoalkaloids, 3 low-molecular-weight amide compounds, and 2 unsaturated fatty acids, including an omega-3 fatty acid. The potato peels showed more potent radical scavenging activity than the flesh. The quantification of the 11 components indicated that the potato peels contained a higher amount of phenolic compounds than the flesh. These results suggest that peel waste from the industry of potato chips and fries may be a source of useful compounds for human health. PMID:22980823

  6. Citric Acid Production by the Aspergillus niger Isolated from the Microflora of Iran

    R.Yazdanparast

    1995-08-01

    Full Text Available Citric acid production by A.niger, isolated from the microflora of Iran, has been investigated in liquid and semi-solid states using growth media with different compositions. In 2% media made of Rocheh grape pomace or sabouraud dextrose, the yield of citric acid production was 0.7 g per Kg of the pomace; and the yield decreased by 50% in 2% saghal solian grape pomace medium. However, in 40% (W/W saghal solian semi-solid medium containing 3% methanol, the yield of citric acid production has improved to 80 g per Kg of pomace in stationary mode of production and to 120 g per Kg of pomace in the rolling mode of fermentation.

  7. Regulation of amino acid transport in isolated rat hepatocytes during development

    Leoni, S.; Spagnuolo, S.; Dini, L.; Devirgiliis, L.C.

    1987-01-01

    The effect of amino acid depletion or supplementation and the effect of glucagon and insulin on the amino acid transport mediated by system A were investigated by determining the uptake of either 2-amino (1-/sup 14/C)isobutyric acid (AIB) or N-methyl 2-amino (1-/sup 14/C)isobutyric acid (MeAIB) in rat hepatocytes, freshly isolated at different stages of pre- and postnatal development. The data obtained show that the Na/sup +/ -dependent uptake was higher at the earliest developmental stages, and steadily decreased until the adult level. The hormones increased AIB and MeAIB uptake enhancing the V/sub max/, while the K/sub m/ was unchanged. This effect was evident in cells from adult and 18-20-day-old fetuses, while no response was present before the 18th day of fetal life and in the prenatal period. Actinomycin D or cycloheximide abolished this hormone-dependent increase. A decrease in AIB and MeAIB transport after incubation in an amino acid-rich medium was demonstrated at all ages tested, but was particularly evident in the prenatal life. The increase in the activity of the system following amino acid starvation was shown to be mostly dependent from de novo protein synthesis in the fetal life; on the contrary in the adult the increase appeared to be more linked to the release from transinhibition of the transport.

  8. Sensitive and selective determining ascorbic acid and activity of alkaline phosphatase based on electrochemiluminescence of dual-stabilizers-capped CdSe quantum dots in carbon nanotube-nafion composite.

    Ma, Xiaolong; Zhang, Xin; Guo, Xinli; Kang, Qi; Shen, Dazhong; Zou, Guizheng

    2016-07-01

    Sensitive and selective determining bio-related molecule and enzyme play an important role in designing novel procedure for biological sensing and clinical diagnosis. Herein, we found that dual-stabilizers-capped CdSe quantum dots (QDs) in composite film of multi-walled carbon nanotubes (CNTs) and Nafion, displaying eye-visible monochromatic electrochemiluminescence (ECL) with fwhm of 37nm, which offers promising ECL signal for detecting ascorbic acid (AA) as well as the activity of alkaline phosphatase (ALP) in biological samples. It was also shown that the dual-stabilizers-capped CdSe QDs can preserve their highly passivated surface states with prolonged lifetime of excited states in Nafion mixtures, and facilitate electron-transfer ability of Nafion film along with CNTs. Compared with the QDs/GCE, the ECL intensity is enhanced 1.8 times and triggering potential shifted to lower energy by 0.12V on the CdSe-CNTs-Nafion/GCE. The ECL quenching degree increases with increasing concentration of AA in the range of 0.01-30nM with a limit of detection (LOD) of 5pM. The activity of ALP was determined indirectly according to the concentration of AA, generated in the hydrolysis reaction of l-ascorbic acid 2-phosphate sesquimagnesium (AA-P) in the presence of ALP as a catalyst, with an LOD of 1μU/L. The proposed strategy is favorable for developing simple ECL sensor or device with high sensitivity, spectral resolution and less electrochemical interference. PMID:27154663

  9. Cloning and sequence analysis of a full-length cDNA of SmPP1cb encoding turbot protein phosphatase 1 beta catalytic subunit

    Qi, Fei; Guo, Huarong; Wang, Jian

    2008-02-01

    Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase 1(PP1cb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPP1cb, by the rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPP1cb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5' untranslated region and 462 bp 3' untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B). And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPP1cb is extremely conserved in both amino acid and nucleotide acid levels compared with the PP1cb of other vertebrates and invertebrates, and its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXX ATGG, which is different from mammalian in two positions A-6 and G-3, indicating the possibility of different initiation of translation in turbot, and also the 3'UTR of SmPP1cb is highly diverse in the sequence similarity and length compared with other animals, especially zebrafish. The cloning and sequencing of SmPP1cb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.

  10. Probing protein phosphatase substrate binding

    Højlys-Larsen, Kim B.; Sørensen, Kasper Kildegaard; Jensen, Knud Jørgen; Gammeltoft, Steen

    2012-01-01

    Proteomics and high throughput analysis for systems biology can benefit significantly from solid-phase chemical tools for affinity pull-down of proteins from complex mixtures. Here we report the application of solid-phase synthesis of phosphopeptides for pull-down and analysis of the affinity...... profile of the integrin-linked kinase associated phosphatase (ILKAP), a member of the protein phosphatase 2C (PP2C) family. Phosphatases can potentially dephosphorylate these phosphopeptide substrates but, interestingly, performing the binding studies at 4 °C allowed efficient binding to phosphopeptides......, without the need for phosphopeptide mimics or phosphatase inhibitors. As no proven ILKAP substrates were available, we selected phosphopeptide substrates among known PP2Cδ substrates including the protein kinases: p38, ATM, Chk1, Chk2 and RSK2 and synthesized directly on PEGA solid supports through a BAL...

  11. Structural Genomics of Protein Phosphatases

    Almo,S.; Bonanno, J.; Sauder, J.; Emtage, S.; Dilorenzo, T.; Malashkevich, V.; Wasserman, S.; Swaminathan, S.; Eswaramoorthy, S.; et al

    2007-01-01

    The New York SGX Research Center for Structural Genomics (NYSGXRC) of the NIGMS Protein Structure Initiative (PSI) has applied its high-throughput X-ray crystallographic structure determination platform to systematic studies of all human protein phosphatases and protein phosphatases from biomedically-relevant pathogens. To date, the NYSGXRC has determined structures of 21 distinct protein phosphatases: 14 from human, 2 from mouse, 2 from the pathogen Toxoplasma gondii, 1 from Trypanosoma brucei, the parasite responsible for African sleeping sickness, and 2 from the principal mosquito vector of malaria in Africa, Anopheles gambiae. These structures provide insights into both normal and pathophysiologic processes, including transcriptional regulation, regulation of major signaling pathways, neural development, and type 1 diabetes. In conjunction with the contributions of other international structural genomics consortia, these efforts promise to provide an unprecedented database and materials repository for structure-guided experimental and computational discovery of inhibitors for all classes of protein phosphatases.

  12. Characteristics of Lactic Acid Bacteria Isolated from Gastrointestinal Tract of Cemani Chicken and Their Potential Use as Probiotics

    S. N. Jannah

    2014-12-01

    Full Text Available The aims of this study were to screen and characterize lactic acid bacteria (LAB isolated from gastrointestinal (GI tract of Cemani chicken, one of Indonesian local chicken and to investigate their potential use as probiotics. LAB were isolated from GI tract using MRSA and GYPA media and incubated anaerobically. Selected LAB were determined their probiotic properties with several assays. Identification of selected LAB was based on 16S rDNA sequences, morphological and biochemical characteristics. Ninety five bacteria were isolated and characterized as lactic acid bacteria (Gram positive, catalase negative, non sporeforming and acid producing. Twenty four isolates of LAB demonstrated antimicrobial activity against Escherichia coli JCM 1649 and Salmonella enteritidis B2586, and three selected isolates, i.e. CCM011, CSP004, and CVM002 showed the highest inhibition activity. The isolates had characters of high cell surface hydrophobicity and inter-isolate coaggregation ability of LAB, high survival at low pH, high phytase and protease activity (but no amylase and lipase activity, weak coaggregation with pathogen and no resistance to the examined antibiotics. The isolates were identified based on sequence analysis of 16S rRNA gene as Lactobacillus salivarius, however, each isolate had different profiles of sugar fermentation. Therefore the three LAB isolates had potential application as probiotics for chicken.

  13. Overexpression of Human Bone Alkaline Phosphatase in Pichia Pastoris

    Karr, Laurel; Malone, Christine, C.; Rose, M. Franklin (Technical Monitor)

    2000-01-01

    The Pichiapastoris expression system was utilized to produce functionally active human bone alkaline phosphatase in gram quantities. Bone alkaline phosphatase is a key enzyme in bone formation and biomineralization, yet important questions about its structural chemistry and interactions with other cellular enzymes in mineralizing tissues remain unanswered. A soluble form of human bone alkaline phosphatase was constructed by deletion of the 25 amino acid hydrophobic C-terminal region of the encoding cDNA and inserted into the X-33 Pichiapastoris strain. An overexpression system was developed in shake flasks and converted to large-scale fermentation. Alkaline phosphatase was secreted into the medium to a level of 32mgAL when cultured in shake flasks. Enzyme activity was 12U/mg measured by a spectrophotometric assay. Fermentation yielded 880mgAL with enzymatic activity of 968U/mg. Gel electrophoresis analysis indicates that greater than 50% of the total protein in the fermentation is alkaline phosphatase. A purification scheme has been developed using ammonium sulfate precipitation followed by hydrophobic interaction chromatography. We are currently screening crystallization conditions of the purified recombinant protein for subsequent X-ray diffraction analyses. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

  14. Probiotic properties of lactic acid bacteria isolated from traditionally fermented Xinjiang cheese.

    Azat, Ramila; Liu, Yan; Li, Wei; Kayir, Abdurihim; Lin, Ding-Bo; Zhou, Wen-Wen; Zheng, Xiao-Dong

    2016-08-01

    Six lactic acid bacterial (LAB) strains were isolated from traditionally fermented Xinjiang cheese and evaluated for functional and probiotic properties and potentials as starter cultures. The isolated six LAB strains comprised Lactobacillus rhamnosus (one strain), Lactobacillus helveticus (one strain), and Enterococcus hirae (four strains). All of the six strains were tolerant to acidic and bile salt conditions. Among which, the L. rhamnosus R4 strain showed more desirable antimicrobial, auto-aggregation, and hydrophobic activity. In addition, the strain L. rhamnosus R4 exhibited the highest level of free radical scavenging activity (53.78% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals and 45.79% of hydroxyl radicals). L. rhamnosus R4 also demonstrated cholesterol and triglyceride degradation by 50.97% and 28.92%, respectively. To further examine the health-promoting effects of these LAB strains on host lifespan, Caenorhabditis elegans was used as an in vivo model. Worms fed LAB as a food source had significant differences in lifespan compared to those fed Escherichia coli OP50 (as a negative control). Feeding of L. rhamnosus R4 extended the mean lifespan of C. elegans by up to 36.1% compared to that of the control. The results suggest that the strains isolated from Xinjiang fermented dairy products have high potential as starter cultures in the cheese industry. PMID:27487805

  15. Isolation of linoleic and alpha-linolenic acids as COX-1 and -2 inhibitors in rose hip

    Jäger, Anna; Petersen, K N; Thomasen, G.;

    2008-01-01

    /2 activity-guided. The bioassay-guided fractionation led to the isolation of linoleic acid (the IC50 for COX-1 was 85 microm and 0.6 microM for COX-2) and alpha-linolenic acid (the IC50 for COX-1 was 52 microM and 12 microM for COX-2). The COX-2/COX-1 ratio was 0.007 for linoleic acid and 0.2 for alpha......-linolenic acid. Linoleic acid and alpha-linolenic acid contribute to the COX-1 and -2 inhibitory activity of rose hip....

  16. Glucose-6-phosphatase deficiency

    Labrune Philippe

    2011-05-01

    Full Text Available Abstract Glucose-6-phosphatase deficiency (G6P deficiency, or glycogen storage disease type I (GSDI, is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, between the ages of 3 to 4 months by symptoms of hypoglycemia (tremors, seizures, cyanosis, apnea. Patients have poor tolerance to fasting, marked hepatomegaly, growth retardation (small stature and delayed puberty, generally improved by an appropriate diet, osteopenia and sometimes osteoporosis, full-cheeked round face, enlarged kydneys and platelet dysfunctions leading to frequent epistaxis. In addition, in GSDIb, neutropenia and neutrophil dysfunction are responsible for tendency towards infections, relapsing aphtous gingivostomatitis, and inflammatory bowel disease. Late complications are hepatic (adenomas with rare but possible transformation into hepatocarcinoma and renal (glomerular hyperfiltration leading to proteinuria and sometimes to renal insufficiency. GSDI is caused by a dysfunction in the G6P system, a key step in the regulation of glycemia. The deficit concerns the catalytic subunit G6P-alpha (type Ia which is restricted to expression in the liver, kidney and intestine, or the ubiquitously expressed G6P transporter (type Ib. Mutations in the genes G6PC (17q21 and SLC37A4 (11q23 respectively cause GSDIa and Ib. Many mutations have been identified in both genes,. Transmission is autosomal recessive. Diagnosis is based on clinical presentation, on abnormal basal values and absence of hyperglycemic response to glucagon. It can be confirmed by demonstrating a deficient activity of a G6P system component in a liver biopsy. To date, the diagnosis is most

  17. Toward Probiotict Food Product from Meat Through Isolation and Identification Lactic Acid Bacteria As Probiotic Culture Stater

    Yunilas Yunilas; Lili Warly; Edi Mirwandhono

    2014-01-01

    Probiotic food products of meat can provide extensive benefits, to increase the shelf life and nutritional value also improve the taste. The use of lactic acid bacteria culture (LAB) derived from the isolation of the meat and the addition of probiotic cultures (Lactobacilli and Bifidobacteria) in fermented sausage processing is expected to produce a probiotic sausage products with nutritional value, and better shelf life and improve health. This study aimed to isolate and identify lactic acid...

  18. Production of conjugated linoleic acids by Lactobacillusplantarum strains isolated from naturally fermented Chinese pickles

    Pei LIU; Sheng-rong SHEN; Hui RUAN; Qian ZHOU; Liu-liu MA; Guo-qing HE

    2011-01-01

    Naturally fermented pickles harbour many lactic acid bacteria (LAB).Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines.Of these isolates,Ip15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 μg/ml of linoleic acid (LA).Compared to other strains,L.plantarum strain Ip15 showed the highest tolerance upon increased levels of LA in the medium,I.e.,up to 600 pg/ml.This strain converted about 25% of LA into CLA isomers [predominantly cis-9,trans-11 CLA (9-CLA) and trans-10,cis-12 CLA (10-CLA)],of which 75% was 9-CLA.Interestingly,though the conversion rate of LA into CLA by Ip15 remained stable between 100 to 600 μg/ml LA levels in the medium,it dropped sharply at 1000 μg/ml.Taken together,the Ip15 strain displayed relatively high LA tolerance with higher conversion rate,which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.

  19. Stereospecific analysis of fatty acid esters of chloropropanediol isolated from fresh goat milk.

    Myher, J J; Kuksis, A; Marai, L; Cerbulis, J

    1986-05-01

    The fatty acid esters of chloropropanediol isolated from goat milk fat in small quantities were subjected to a stereospecific analysis via phospholipase C and phosphocholine esters as intermediates. Synthetic rac-1-chloro-2,3-dioleoyl-propanediol was prepared by standard methods and was used as a control. The stereospecific analyses were performed following a release of the fatty acids from the primary positions of each chloropropanediol diester with pancreatic lipase. The resulting X-1-chloro-2-acylpropanediols were then converted into the corresponding phosphocholine derivatives by a stepwise reaction with phosphorus oxychloride and choline chloride. The X-1-chloro-2-acyl-3-phosphocholinepropanediols were subjected to hydrolysis with phospholipase C (C. perfringens), which hydrolyzed 50% of the phosphatide within two min and the rest of it in two hr. From previous experience with glycerol esters, it was assumed that the more rapidly hydrolyzed molecules were the sn-1-chloro-2-acyl-propanediol derivatives and the more slowly hydrolyzed ones the sn-2-acyl-3-chloropropanediol derivatives. A hydrolysis with phospholipase A2 (Crotalus adamanteus) released 50% of the total fatty acid along with the corresponding lyso compound within 10 min, after which there was no further reaction. The hydrolysis products were assayed directly by gas liquid chromatography (GLC) or were isolated by thin layer chromatography (TLC) prior to quantitation by GLC. Both naturally occurring and synthetic chloropropanediol diesters behaved similarly on stereospecific analysis and were therefore concluded to be racemic. PMID:3724368

  20. Efficient isolation of high quality nucleic acids from different tissues of Taxus baccata L.

    Abbasi Kejani, Abolghasem; Hosseini Tafreshi, Sayed Ali; Khayyam Nekouei, Sayed Mojtaba; Mofid, Mohammad Reza

    2010-02-01

    Improved and efficient methods were developed for isolating high quality DNA and RNA from different sources of Iranian Yew (Taxus baccata L.). The methods were based on CTAB extraction buffer added with high levels of polyvinylpyrrolidone (PVP) and beta-mercaptoethanol to properly remove polysaccharides and prevent oxidation of phenolics. The pellets obtained by ethanol precipitation were washed only with Chloroform: isoamyl alcohol (24:1). So, we could successfully eliminate the dangerous phenol/chloroform extraction steps from the isolation procedure. Both spectrophotometric (A(260)/A(280) and A(260)/A(230) ratios) and agarose electrophoresis analysis of isolated nucleic acids (DNA and RNA) indicated good results. DNA with the average yield of 100-300 microg/g leaf and stem tissue and total RNA with an average yield of 20-30 microg/g cell culture and 80-100 microg/g leaf and stem tissue of Iranian yew could be obtained. Successful amplification of pam and pds by PCR and RT-PCR, showed the integrity of isolated DNA and RNA, respectively. PMID:19578976

  1. Isolation and Selection of Anti-Candida albicans Producing Lactic Acid Bacteria

    Monthon LERTCANAWANICHAKUL

    2005-06-01

    Full Text Available The forty isolates of lactic acid bacteria (LAB were obtained from various fermented foods. The cross streak plate method was used to preliminary screen for antimicrobial activity. LAB were isolated by selective medium, Mann Rogosa Sharpe (MRS. Most of the isolates showed inhibition against Staphylococcus aureus TISTR 517, Bacillus subtilis TISTR 008, Micrococcus luteus TISTR 884, Escherichia coli TISTR 887, Pseudomonas aeruginosa TISTR 781, and Candida albicans DMST 5239. Only sterile culture supernatant of isolate No. L14, later identified as Lactococcus lactis, showed antifungal activity by means of agar well diffusion assay. The activity was stable during heat treatment and was retained even after autoclaving at 121 °C for 15 minutes. Maximum activity was observed at pH values between 2.5-4.0, and was lost at higher pH values. The anti-C. albicans activity was fully regained after readjustment of the pH to the initial value (pH 3.5.

  2. Enterococcus bulliens sp. nov., a novel lactic acid bacterium isolated from camel milk.

    Kadri, Zaina; Spitaels, Freek; Cnockaert, Margo; Praet, Jessy; El Farricha, Omar; Swings, Jean; Vandamme, Peter

    2015-11-01

    Four lactic acid bacteria isolates obtained from fresh dromedary camel milk produced in Dakhla, a city in southern Morocco, were characterised in order to determine their taxonomic position. The four isolates had highly similar MALDI-TOF MS and RAPD fingerprints and identical 16S rRNA gene sequences. Comparative sequence analysis revealed that the 16S rRNA gene sequence of the four isolates was most similar to that of Enterococcus sulfureus ATCC 49903(T) and Enterococcus italicus DSM 15952(T) (99.33 and 98.59% similarity, respectively). However, sequence analysis of the phenylalanyl-tRNA synthase (pheS), RNA polymerase (rpoA) and ATP synthase (atpA) genes revealed that the taxon represented by strain LMG 28766(T) was well separated from E. sulfureus LMG 13084(T) and E. italicus LMG 22039(T), which was further confirmed by DNA-DNA hybridization values that were clearly below the species demarcation threshold. The novel taxon was easily differentiated from its nearest neighbour species through sequence analysis of protein encoding genes, MALDI-TOF mass spectrometry and multiple biochemical tests, but had a similar percentage G+C content of about 39%. We therefore propose to formally classify these isolates as Enterococcus bulliens sp. nov., with LMG 28766(T) (=CCMM B1177(T)) as the type strain. PMID:26346480

  3. Anxiolytic potential of ursolic acid derivative-a stearoyl glucoside isolated from Lantana camara L. (verbanaceae)

    Imran Kazmi; Muhammad Afzal; Babar Ali; Zoheir A Damanhouri; Aftab Ahmaol; Firoz Anwar

    2013-01-01

    Objective: To investigate the anxiolytic activity of newly isolated compound by our lab called ursolic acid stearoyl glucoside (UASG) from the leaves of Lantana camara (L. camara). Methods:Column chromatography was used to isolate UASG. Anxiolytic potential was experimentally proved and demonstrated through Elevated plus-maze, Open field and light and dark test.Results:The UASG showed marked increased in time spent (%) and number of frequent movements made by animals in open arm of elevated plus-maze apparatus. In light and dark model, UASG produced marked increase in time spent by animal, number of crossing and reduced duration of immobility in light box. Conclusions: UASG showed significant increase in number of rearing, assisted rearing and number of square crossed in open field established test model. UASG showed its anxiolytic effect in dose dependent manner.

  4. The effect of cigarette smoke on the metabolism of arachidonic acid in isolated hamster lungs

    The effects of cigarette smoke on the metabolism of exogenous arachidonic acid (AA) were investigated in isolated hamster lungs. Arachidonate was injected into the pulmonary circulation and the metabolites were analysed from the nonrecirculating perfusion effluent by thin layer chromatography. After the pulmonary injection of 66 nmol of 14C-AA about 20% of the injected radioactivity appeared in the perfusion effluent mostly as metabolites in six minutes. When isolated lungs were ventilated with cigarette smoke during the perfusion, the amounts of PGF2 alpha, PGE2 and two unidentified metabolite groups increased in the lung effluent. In two other experimental series hamsters were exposed to cigarette smoke before the lung perfusion either once for 30 min or during one hour daily for ten consecutive days. Neither pre-exposures caused any changes in the amounts of arachidonate metabolites in the lung effluent

  5. Dephosphorylation of chicken cardiac myofibril C-protein by protein phosphatases 1 and 2A

    C-Protein, which is a regulatory component of cardiac muscle myofibrils, is phosphorylated in response to β-adrenergic agonists by a cAMP-dependent mechanism and dephosphorylated in response to cholinergic agonists. It is believed that the cAMP-dependent phosphorylation is due to cAMP-dependent protein kinase. The protein phosphatase(s) involved in the dephosphorylation of C-protein has not been determined. In this study, chicken cardiac C-protein was phosphorylated with the cAMP-dependent protein kinase to about 3 mol phosphate/mol C-protein. Incubation of [32P]C-protein with the catalytic subunit of protein phosphatase 1 or 2A rapidly removed 30-40% of 32[P]. Phosphopeptide maps and phosphoamino acid analysis revealed that the major site(s) dephosphorylated by either phosphatase was a phosphothreonine residue(s) located on the same tryptic peptide and on the same CNBr fragment. Increasing the incubation period or the phosphatase concentration did not result in any further dephosphorylation of C-protein by phosphatase 1, but phosphatase 2A completely dephosphorylated C-protein. Preliminary studies showed that the major protein phosphatase associated with the myofibril was phosphatase 2A. These results indicate the phosphatase 2A may be important in the regulation of the phosphorylation state of C-protein

  6. Characterization, Identification and Application of Lactic Acid Bacteria Isolated from Forage Paddy Rice Silage

    Ni, Kuikui; Wang, Yanping; Li, Dongxia; Cai, Yimin; Pang, Huili

    2015-01-01

    There has been growing interest to develop forage rice as a new feed resource for livestock. This study was to characterize the natural population of lactic acid bacteria (LAB) and select potentially excellent strains for paddy rice silage preparation in China. One hundred and twenty-six strains were isolated and screened from paddy rice silage prepared using a small-scale fermentation system, and ninety-nine of these isolates were considered to be LAB based on their Gram-positive and catalase-negative morphology and the production of most of their metabolic products as lactic acid. These isolates were divided into eight groups (A-H) on the basis of their morphological and biochemical characteristics. The Group A to H strains were identified as Lactobacillus (L.) plantarum subsp. plantarum (species ratio: 8.1%), L. casei (5.1%), Leuconostoc (Ln.) pseudomesenteroides (11.1%), Pediococcus (P.) pentosaceus (24.2%), Enterococcus (E.) mundtii (12.1%), Lactococcus (Lc.) garvieae (15.2%), E. faecium (9.1%) and Lc. lactis subsp. lactis (15.2%) based on sequence analyses of their 16S rRNA and recA genes. P. pentosaceus was the most abundant member of the LAB population in the paddy rice silage. A selected strain, namely L. casei R 465, was found to be able to grow under low pH conditions and to improve the silage quality with low pH and a relatively high content of lactic acid. This study demonstrated that forage paddy rice silage contains abundant LAB species and its silage can be well preserved by inoculation with LAB, and that strain R 465 can be a potentially excellent inoculant for paddy rice silage. PMID:25803578

  7. Isolation of a foot-and-mouth disease polyuridylic acid polymerase and its inhibition by antibody.

    Polatnick, J

    1980-01-01

    A template-dependent polyuridylic acid [poly(U)] polymerase has been isolated from BHK cells infected with foot-and-mouth disease virus (FMDV). Enzyme activity in a 20,000 x g supernatant of a cytoplasmic extract was concentrated by precipitation with 30 to 50% saturated ammonium sulfate. The poly(U) polymerase was freed of membranes by sodium dodecyl sulfate and 1,1,2-trichlorotrifluoroethane extraction, and RNA was removed by precipitation with 2 M LiCl. The solubilized poly(U) polymerase r...

  8. Isolation and fractionation of soil humin using alkaline urea and dimethylsulphoxide plus sulphuric acid

    Song, Guixue; Hayes, Michael H. B.; Novotny, Etelvino H.; Simpson, Andre J.

    2011-01-01

    Humin, the most recalcitrant and abundant organic fraction of soils and of sediments, is a significant contributor to the stable carbon pool in soils and is important for the global carbon budget. It has significant resistance to transformations by microorganisms. Based on the classical operational definition, humin can include any humic-type substance that is not soluble in water at any pH. We demonstrate in this study how sequential exhaustive extractions with 0.1 M sodium hydroxide (NaOH) + 6 M urea, followed by dimethylsulphoxide (DMSO) + 6% ( v/ v) sulphuric acid (H2SO4) solvent systems, can extract 70-80% of the residual materials remaining after prior exhaustive extractions in neutral and aqueous basic media. Solid-state 13C NMR spectra have shown that the components isolated in the base + urea system were compositionally similar to the humic and fulvic acid fractions isolated at pH 12.6 in the aqueous media. The NMR spectra indicated that the major components isolated in the DMSO + H2SO4 medium had aliphatic hydrocarbon associated with carboxyl functionalities and with lesser amounts of carbohydrate and peptide and minor amounts of lignin-derived components. The major components will have significant contributions from long-chain fatty acids, waxes, to cuticular materials. The isolates in the DMSO + H2SO4 medium were compositionally similar to the organic components that resisted solvation and remained associated with the soil clays. It is concluded that the base + urea system released humic and fulvic acids held by hydrogen bonding or by entrapment within the humin matrix. The recalcitrant humin materials extracted in DMSO + H2SO4 are largely biological molecules (from plants and the soil microbial population) that are likely to be protected from degradation by their hydrophobic moieties and by sorption on the soil clays. Thus, the major components of humin do not satisfy the classical definitions for humic substances which emphasise that these arise from

  9. Anti-listerial inhibitory lactic acid bacteria isolated from commercial cold smoked salmon

    Tome, Elisabetta; Teixeira, Paula; Gibbs, Paul A.

    2006-01-01

    The natural microflora of cold-smoked fish at the end of shelf-life are lactic acid bacteria (LAB). Some of these display a capacity to inhibit spoilage as well as several strains of pathogenic micro-organisms, e.g. Listeria monocytogenes which is isolated frequently from cold-smoked salmon (CSS). Eight batches of sliced vacuum-packed CSS from Norway, Scotland and Spain were collected at retail. Packs were stored at 5 1C and examined for chemical and microbiological characteristic...

  10. Lactic acid bacteria from Jijel's traditional butter: Isolation, identification and major technological traits

    Idoui, Tayeb

    2008-12-01

    Full Text Available Twenty seven (27 lactic acid bacteria were isolated from Jijel’s traditional butter. These strains belong to three genera: Lactococcus, Lactobacillus and Leuconostoc. The results showed that Lactobacillus plantarum was the predominant species in this traditional butter. It appears that these strains have some interesting technological properties.Se aíslan veintisiete (27 bacterias acidolácticas de la mantequilla tradicional de Jijel. Éstas pertenecen a los géneros Lactococcus, Lactobacillus y Leuconostoc. Los resultados muestran que Lactobacillus plantarum es la especie predominante en dicha mantequilla. Diversas cepas presentan algunas propiedades tecnológicas interesantes.

  11. Catabolism of 15-(p-iodophenyl)-R,S-β-methylpentadecanoic acid (BMIPP) by isolated rat hearts

    The aim of the studies with radioiodinated long chain fatty acids was to investigate their use as potential probes for β-oxidation, or net energy-producing LCFA utilization, because such probes would be useful in studies of cardiac biochemistry and its derangement in conditions such as ischaemic heart disease. A major result of the work in the isolated working rat heart is that BMIPP kinetics is insensitive to the large decline in β-oxidation rate induced by the carnitine palmitoyltransferase I (CPT I) inhibitor 2-[5-(4-chlorophenyl)-pentyl]oxirane-2-carboxylate (POCA). (orig./MG)

  12. Behavior of Psychrotrophic Lactic Acid Bacteria Isolated from Spoiling Cooked Meat Products

    Hamasaki, Yoshikatsu; Ayaki, Mitsuko; Fuchu, Hidetaka; Sugiyama, Masaaki; Morita, Hidetoshi

    2003-01-01

    Three kinds of lactic acid bacteria were isolated from spoiling cooked meat products stored below 10°C. They were identified as Leuconostoc mesenteroides subsp. mesenteroides, Lactococcus lactis subsp. lactis, and Leuconostoc citreum. All three strains grew well in MRS broth at 10°C. In particular, L. mesenteroides subsp. mesenteroides and L. citreum grew even at 4°C, and their doubling times were 23.6 and 51.5 h, respectively. On the other hand, although the bacteria were initially below the...

  13. Selection of exopolysaccharide-producing lactic acid bacteria isolates from Inner Mongolian traditional yoghurt

    Zhang Chun-lei

    2014-11-01

    Full Text Available Lactic acid bacteria (LAB isolated from Inner Mongolian traditional yoghurt were evaluated for the production of exopolysaccharides (EPS by phenol-sulphuric acid method after ethanol precipitation and dialysis. Total polysaccharide was extracted from sucrose-containing MRS broth cultures of the selected LAB strains. Comparison of the EPS yields revealed that among tested LAB, strain 37 exhibited the highest production of 536.904 mg/L. The strain was identified as Leuconostoc citreum with carbohydrate assimilation profiling, 16S rRNA and pheS gene sequencing. The Ln. citreum 37 was found to be a novel EPS producing strain. It was found that there was no direct linear relation between the colony size and EPS yield, so the colony size could not to be used to screen EPS-producing strains.

  14. Nanocellulose prepared by acid hydrolysis of isolated cellulose from sugarcane bagasse

    Wulandari, W. T.; Rochliadi, A.; Arcana, I. M.

    2016-02-01

    Cellulose in nanometer range or called by nano-cellulose has attracted much attention from researchers because of its unique properties. Nanocellulose can be obtained by acid hydrolysis of cellulose. The cellulose used in this study was isolated from sugarcane bagasse, and then it was hydrolyzed by 50% sulfuric acid at 40 °C for 10 minutes. Nanocellulose has been characterized by Transmission Electron Microscope (TEM), Particle Size Analyzer (PSA), Fourier Transform Infrared Spectroscopy (FTIR) and X-Ray Diffraction (XRD). Analysis of FTIR showed that there were not a new bond which formed during the hydrolysis process. Based on the TEM analysis, nano-cellulose has a spherical morphology with an average diameter of 111 nm and a maximum distribution of 95.9 nm determined by PSA. The XRD analysis showed that the crystallinity degree of nano-cellulose was higher than cellulose in the amount of 76.01%.

  15. Effects of tumour mass and circulating antigen on the biodistribution of 111In-labelled F(ab')2 fragments of human prostatic acid phosphatase monoclonal antibody in nude mice bearing PC-82 human prostatic tumor xenografts

    We have evaluated the effects of tumour mass and circulating antigen (prostatic acid phosphatase, PAP) on the biodistribution and the incorporation of 111In-labelled F(ab')2 monoclonal antibody (MoAb) fragments directed against human PAP into human prostatic tumours (PC-82; 0.1-8.9 g) growing in nude mice. The radioactivities in the blood, liver, spleen, kidney and tumour were compared at 1, 3, 4 and 6 days after the intravenous administration of the antibody fragments. There was a significant correlation between the tumour size and the serum PAP concentration in the model employed. Even tissue of a small tumour (111In-labelled F(ab')2 fragments. This relationship had levelled off by 72 h and most likely reflected a better vascularisation of the smaller tumours. Our results show that the increase in tumour size and in the concentration of circulating antigen in the blood led to decreased tumour-to-blood ratios, since there was a tendency for higher blood activities in mice with larger tumours and higher serum PAP concentrations. There was no correlation between tumour size and label uptake by the liver during the follow-up over 144 h, although serum PAP concentrations ranged from 3.1 μg/l to 352 μg/l. On the other hand, when compared with our previous data obtained with non-tumour-bearing mice, there was a significant increase in the uptake by the liver and spleen. These results indicate that even a small concentration of circulating antigen was able to trigger an abnormal change in the biodistribution of MoAbs. (orig.)

  16. New method for the rapid extraction of natural products: efficient isolation of shikimic acid from star anise.

    Just, Jeremy; Deans, Bianca J; Olivier, Wesley J; Paull, Brett; Bissember, Alex C; Smith, Jason A

    2015-05-15

    A new, practical, rapid, and high-yielding process for the pressurized hot water extraction (PHWE) of multigram quantities of shikimic acid from star anise (Illicium verum) using an unmodified household espresso machine has been developed. This operationally simple and inexpensive method enables the efficient and straightforward isolation of shikimic acid and the facile preparation of a range of its synthetic derivatives. PMID:25938329

  17. Uptake of ascorbic acid by freshly isolated cells and secretory granules from the intermediate lobe of ox hypophyses

    Zhou, A; Matsumoto, T; Farver, O;

    1990-01-01

    Mechanically isolated cells from the intermediate lobe of ox hypophyses contained 40.6 +/- 3.7 nmol mg-1 protein (mean +/- SE, n = 5) of ascorbic acid. They accumulated radioactivity time dependently, on incubation with L-[14C]ascorbic acid in ionic medium dominated by NaCl. No definite saturation...

  18. Draft Genome Sequences of Gluconobacter cerinus CECT 9110 and Gluconobacter japonicus CECT 8443, Acetic Acid Bacteria Isolated from Grape Must

    Sainz, Florencia

    2016-01-01

    We report here the draft genome sequences of Gluconobacter cerinus strain CECT9110 and Gluconobacter japonicus CECT8443, acetic acid bacteria isolated from grape must. Gluconobacter species are well known for their ability to oxidize sugar alcohols into the corresponding acids. Our objective was to select strains to oxidize effectively d-glucose. PMID:27365351

  19. Cholesterol assimilation by lactic acid bacteria and bifidobacteria isolated from the human gut.

    Pereira, Dora I A; Gibson, Glenn R

    2002-09-01

    The objective of this study was to evaluate the effect of human gut-derived lactic acid bacteria and bifidobacteria on cholesterol levels in vitro. Continuous cultures inoculated with fecal material from healthy human volunteers with media supplemented with cholesterol and bile acids were used to enrich for potential cholesterol assimilators among the indigenous bacterial populations. Seven potential probiotics were found: Lactobacillus fermentum strains F53 and KC5b, Bifidobacterium infantis ATCC 15697, Streptococcus bovis ATCC 43143, Enterococcus durans DSM 20633, Enterococcus gallinarum, and Enterococcus faecalis. A comparative evaluation regarding the in vitro cholesterol reduction abilities of these strains along with commercial probiotics was undertaken. The degree of acid and bile tolerance of strains was also evaluated. The human isolate L. fermentum KC5b was able to maintain viability for 2 h at pH 2 and to grow in a medium with 4,000 mg of bile acids per liter. This strain was also able to remove a maximum of 14.8 mg of cholesterol per g (dry weight) of cells from the culture medium and therefore was regarded as a candidate probiotic. PMID:12200334

  20. Effect of lactic acid bacteria isolated from fermented mustard on immunopotentiating activity

    Chen-Kai; Chang; Shu-Chen; Wang; Chih-Kwang; Chiu; Shih-Ying; Chen; Zong-Tsi; Chen; Pin-Der; Duh

    2015-01-01

    Objective: To investigate the effect of lactic acid bacteria isolated from fermented mustard on immunopotentiating activity Methods: One hundred and fifty nine strains of lactic acid bacteria isolated from traditional Taiwan fermented mustard were evaluated for their immunopotentiating activity on a murine macrophage cell line RAW 264.7.Results: Of the strains, pronounced increases in the levels of nitric oxide(NO), tumor necrosis factor-α and interleukin-6 were observed in strains B0040, B0110 and B0145. Among them,strain B0145 had the highest NO and tumor necrosis factor-α generation in RAW 264.7 cells;strains B0040 and B0110 were also superior to that of Lactobacillus casei. These results demonstrated that NO and cytokines were effectively induced when the bacterial stimulants were treated with macrophages. In addition, strains B0040 and B0110 were identified as Lactobacillus plantarum, and B0145 as Weissella cibaria using 16 S rDNA analysis.Conclusions: The results implicated selected strains may be regarded as a biological response modifier and had a broad application prospects in exploiting new functional food or as a feed additive.

  1. Effect of Lanthanum on Acid Secretion from Isolated Mouse Stomach in Vitro

    徐项桂; 夏洪涛; 芮光; 胡翠英; 袁福根

    2004-01-01

    To explore the effect and the mechanism of La3+ on gastric acid secretion (GAS) of isolated mouse stomach with perfused lumen, 12 cm H2O column intragastric pressure-provided, whole stomach preparations from mice were incubated in buffer at 37 ℃ in vitro, and perfusate was measured for pH with a pHS-3 type pH meter. The results show that La3+ (0.41~820×10-6 mol*L-1) significantly promotes GAS in a concentration-dependant manner. Proglutamine, a blocker of gastrin receptor, potently inhibits GAS, and it may block the promotive effect of La3+ on GAS, and this effect increases with the increase of proglutamin concentration. Cimetidine, a blocker of histamine H2 receptor, also potently inhibits GAS, and blocks the promotive effect of La3+ on GAS in the same manner with proglutamine. These results suggest that La3+ promotes GAS in isolated stomach possibly by stimulating the releases of gastrin from G cell and Histamine from ECL cell or by activating the gastrin receptors and Histamine H2 receptors on the parietal cell, thereby accelerating the acid secretion of parietal cells in stomach.

  2. Novel Simplified and Rapid Method for Screening and Isolation of Polyunsaturated Fatty Acids Producing Marine Bacteria

    Ashwini Tilay

    2012-01-01

    Full Text Available Bacterial production of polyunsaturated fatty acids (PUFAs is a potential biotechnological approach for production of valuable nutraceuticals. Reliable method for screening of number of strains within short period of time is great need. Here, we report a novel simplified method for screening and isolation of PUFA-producing bacteria by direct visualization using the H2O2-plate assay. The oxidative stability of PUFAs in growing bacteria towards added H2O2 is a distinguishing characteristic between the PUFAs producers (no zone of inhibition and non-PUFAs producers (zone of inhibition by direct visualization. The confirmation of assay results was performed by injecting fatty acid methyl esters (FAMEs produced by selected marine bacteria to Gas Chromatography-Mass Spectrometry (GCMS. To date, this assay is the most effective, inexpensive, and specific method for bacteria producing PUFAs and shows drastically reduction in the number of samples thus saves the time, effort, and cost of screening and isolating strains of bacterial PUFAs producers.

  3. Naturally fermented Jijelian black olives: microbiological characteristics and isolation of lactic acid bacteria

    Karam, Nour-Eddine

    2009-12-01

    Full Text Available A study of the microflora of traditionally fermented black olives in Eastern Algeria is presented. A count of the following microbial groups was carried out: mesophilic bacteria, enterobacteria, lactic acid bacteria (LAB, staphylococci and yeast. In a second phase, the identification and assessment of the technological traits of LAB was performed. Seventeen lactic acid bacteria were isolated and identified. These isolates were represented by two genera: Lactobacillus and Leuconostoc. The results showed that Lactobacillus plantarum was the predominant species in this traditional product.Un estudio sobre la microflora de aceitunas negras fermentada por métodos tradicionales en el Este de Argelia es presentado. Se realizo el siguiente recuento de grupos de microorganismos: bacterias mesófilas, enterobacterias, bacterias ácido lácticas (LAB, staphylococcus y levaduras. En una segunda fase, la identificación y evaluación de aspectos tecnológicos de LAB fue realizada. Setenta bacterias ácido lácticas fueron aisladas e identificadas. Estos aislados contenían principalmente dos géneros: Lactobacillus y Leuconostoc. Los resultados mostraron que Lactobacillus plantarum fue la especie predominante en este producto tradicional.

  4. Biodegradation of dimethyl phthalate by Sphingomonas sp. isolated from phthalic-acid-degrading aerobic granules.

    Zeng, Ping; Moy, Benjamin Yan-Pui; Song, Yong-Hui; Tay, Joo-Hwa

    2008-10-01

    Phthalic acid esters (PAEs) contamination in water, air, and soil is one of the major environmental concerns in many countries. Besides the PAE biodegradation process, the PAE degrading bacteria have become one of the focuses of study. This study reports the successful isolation of one kind of indigenous bacterium PA-02 from phthalic acid (PA)-degrading aerobic granules. Based on its 16S ribosomal DNA sequence, isolate PA-02 was identified as Sphingomonas genus with 100% similarity to Sphingomonas sp. strain D84532. Strain PA-02 was a Gram-negative, rod-shaped bacterium with strong auto-aggregation ability. In particular, the strain PA-02 possessed PAE-degrading ability without acclimation. Results of growth tests showed that strain PA-02 could degrade dimethyl phthalate (DMP), dibutyl phthalate, and diethylhexyl phthalate. The specific degradation rates of DMP and PA were concentration-dependent with maximum values of 0.4 g-DMP g(-1) biomass h(-1) and 1.3 g-PA g(-1) biomass h(-1), respectively. Kinetic studies also revealed that PA-02 was robust under high concentrations of DMP and PA. Even when the PA concentration was increased to 1,000.0 mg l(-1), the specific PA degradation rate was about 0.25 g-PA g(-1) biomass h(-1). The corresponding value for DMP was 0.067 g-DMP g(-1) biomass h(-1) at 1,000 mg l(-1). PMID:18751698

  5. Indole Acetic Acid Production by the Indigenous Isolates of Azotobacter and Fluorescent Pseudomonas in the Presence and Absence of Tryptophan

    Ahmad, Farah; Ahmad, Iqbal; KHAN, Mohd Saghir

    2005-01-01

    A total of 21 bacterial isolates (Azotobacter sp., 10 and fluorescent Pseudomonas sp., 11) were isolated from different rhizospheric soils in the vicinity of Aligarh city and characterized as per standard methods. These isolates were further tested for the production of indole acetic acid (IAA) in a medium with 0, 1, 2 and 5 mg/ml of tryptophan. A low amount (2.68-10.80 mg/ml) of IAA production was recorded by Azotobacter strains without tryptophan addition. Seven Azotobacter isolates showed ...

  6. Isolation of chlorogenic acid from Mutellina purpurea L. herb using high-performance counter-current chromatography.

    Sieniawska, Elwira; Skalicka-Woźniak, Krystyna

    2014-01-01

    The aim of the study was to explore proper isolation conditions of chlorogenic acid from the herb of Mutelina purpurea L. - a new source of this bioactive molecule. The accelerated solvent extraction (ASE) with 40% aqueous solution of methanol combined with high-performance counter-current chromatography (HPCCC) was utilised for the efficient extraction and the separation of chlorogenic acid from the M. purpurea herb in less than 30 min. The structure of the obtained compound was confirmed by mass spectrometry and NMR analysis. The preparative HPCCC was performed using the mixture of ethyl acetate, butanol and water (4:1:5, v/v/v) in the reverse-phase mode. The chlorogenic acid was isolated from this herb for the first time, yielding 96% purity. The ASE with 40% methanol combined with HPCCC separation was proven to be a useful tool for quick and efficient isolation of chlorogenic acid from M. purpurea. PMID:25185707

  7. Isolation and characterization of fatty acid methyl ester (FAME)-producing Streptomyces sp. S161 from sheep (Ovis aries) faeces.

    Lu, Y; Wang, J; Deng, Z; Wu, H; Deng, Q; Tan, H; Cao, L

    2013-09-01

    An actinomycete producing oil-like mixtures was isolated and characterized. The strain was isolated from sheep faeces and identified as Streptomyces sp. S161 based on 16S rRNA gene sequence analysis. The strain showed cellulase and xylanase activities. The (1) H nuclear magnetic resonance (NMR) spectra of the mixtures showed that the mixtures were composed of fatty acid methyl esters (52·5), triglycerides (13·7) and monoglycerides (9·1) (mol.%). Based on the gas chromatography-mass spectrometry (GC-MS) analysis, the fatty acid methyl esters were mainly composed of C14-C16 long-chain fatty acids. The results indicated that Streptomyces sp. S161 could produce fatty acid methyl esters (FAME) directly from starch. To our knowledge, this is the first isolated strain that can produce biodiesel (FAME) directly from starch. PMID:23692633

  8. Determination of trace alkaline phosphatase by affinity adsorption solid substrate room temperature phosphorimetry based on wheat germ agglutinin labeled with 8-quinolineboronic acid phosphorescent molecular switch and prediction of diseases

    Liu, Jia-Ming; Gao, Hui; Li, Fei-Ming; Shi, Xiu-Mei; Lin, Chang-Qing; Lin, Li-Ping; Wang, Xin-Xing; Li, Zhi-Ming

    2010-09-01

    The 8-quinolineboronic acid phosphorescent molecular switch (abbreviated as PMS-8-QBA. Thereinto, 8-QBA is 8-quinolineboronic acid, and PMS is phosphorescent molecular switch) was found for the first time. PMS-8-QBA, which was in the "off" state, could only emit weak room temperature phosphorescence (RTP) on the acetyl cellulose membrane (ACM). However, PMS-8-QBA turned "on" automatically for its changed structure, causing that the RTP of 8-QBA in the system increased, after PMS-8-QBA-WGA (WGA is wheat germ agglutinin) was formed by reaction between -OH of PMS-8-QBA and -COOH of WGA. More interesting is that the -NH 2 of PMS-8-QBA-WGA could react with the -COOH of alkaline phosphatase (AP) to form the affinity adsorption (AA) product WGA-AP-WGA-8-QBA-PMS (containing -NH-CO- bond), which caused RTP of the system to greatly increase. Thus, affinity adsorption solid substrate room temperature phosphorimetry using PMS-8-QBA as labelling reagent (PMS-8-QBA-AA-SSRTP) for the determination of trace AP was established. The method had many advantages, such as high sensitivity (the detection limit (LD) was 2.5 zg spot -1. For sample volume of 0.40 μl spot -1, corresponding concentration was 6.2 × 10 -18 g ml -1), good selectivity (the allowed concentration of coexisting material was higher, when the relative error was ±5%), high accuracy (applied to detection of AP content in serum samples, the result was coincided with those obtained by enzyme-linked immunoassay), which was suitable for the detection of trace AP content in serum samples and the forecast of human diseases. Meanwhile, the mechanism of PMS-8-QBA-AASSRTP was discussed. The new field of analytical application and clinic diagnosis technique of molecule switch are exploited, based on the phosphorescence characteristic of PMS-8-QBA, the AA reaction between WGA and AP, as well as the relation between AP content and human diseases. The research results promote the development and interpenetrate among molecule

  9. Characterization of lactic acid bacteria isolated from Bukuljac, a homemade goat's milk cheese.

    Nikolic, Milica; Terzic-Vidojevic, Amarela; Jovcic, Branko; Begovic, Jelena; Golic, Natasa; Topisirovic, Ljubisa

    2008-02-29

    The Bukuljac cheese is traditionally homemade cheese, produced from heat-treated goat's milk without the addition of any bacterial starter culture. The presence of lactic acid bacteria (LAB) in Bukuljac cheese has been analyzed by using a polyphasic approach including microbiological and molecular methods such as rep-PCR with (GTG)5 primer. Lactobacillus paracasei subsp. paracasei represents a dominant strain in the microflora of analyzed cheese. Out of 55 Gram-positive and catalase-negative isolates, 48 belonged to L. paracasei subsp. paracasei species. Besides lactobacilli, five Lactococcus lactis subsp. lactis and two Enterococcus faecalis were found. Results of PCR-denaturing gradient gel electrophoresis (DGGE) of DNA extracted directly from the fresh cheese revealed the presence of Leuconostoc mesenteroides. Only lactobacilli showed a high proteolytic activity and hydrolyzed alpha(s1)- and beta-caseins. They are also producers of diacetyl. In addition, 34 out of 55 isolates, all determined as lactobacilli, showed the ability of auto-aggregation. Among 55 isolates, 50 also exhibited antimicrobial activity. PMID:18177967

  10. Inhibition of Paenibacillus larvae by lactic acid bacteria isolated from fermented materials.

    Yoshiyama, Mikio; Wu, Meihua; Sugimura, Yuya; Takaya, Noriko; Kimoto-Nira, Hiromi; Suzuki, Chise

    2013-01-01

    We evaluated the potential application of lactic acid bacteria (LAB) isolated from fermented feeds and foods for use as probiotics against Paenibacillus larvae, the causal agent of American foulbrood (AFB) in vitro. We also assessed the ability of LAB to induce the expression of antimicrobial peptide genes in vivo. Screening of the 208 LAB isolated from fermented feeds and foods revealed that nine strains inhibited the in vitro growth of P. larvae. The LAB strains were identified by 16S rRNA gene sequencing as Enterococcus sp., Weissella sp. and Lactobacillus sp. These strains were screened for their abilities of immune activation in honeybees by real-time RT-PCR using antimicrobial peptide genes as markers. After oral administration of several of the screened LAB to larvae and adults, the transcription levels of antimicrobial peptide genes, such as abaecin, defensin and hymenoptaecin, were found to increase significantly. These findings suggested that selected LAB stimulate the innate immune response in honeybees, which may be useful for preventing bacterial diseases in honeybees. This is the first report to characterize the probiotic effects of LAB isolated from fermented feeds and foods in honeybees. PMID:23000777

  11. MAP kinase phosphatase 2 regulates macrophage-adipocyte interaction.

    Huipeng Jiao

    Full Text Available Inflammation is critical for the development of obesity-associated metabolic disorders. This study aims to investigate the role of mitogen-activated protein kinase phosphatase 2 (MKP-2 in inflammation during macrophage-adipocyte interaction.White adipose tissues (WAT from mice either on a high-fat diet (HFD or normal chow (NC were isolated to examine the expression of MKP-2. Murine macrophage cell line RAW264.7 stably expressing MKP-2 was used to study the regulation of MKP-2 in macrophages in response to saturated free fatty acid (FFA and its role in macrophage M1/M2 activation. Macrophage-adipocyte co-culture system was employed to investigate the role of MKP-2 in regulating inflammation during adipocyte-macrophage interaction. c-Jun N-terminal kinase (JNK- and p38-specific inhibitors were used to examine the mechanisms by which MKP-2 regulates macrophage activation and macrophage-adipocytes interaction.HFD changed the expression of MKP-2 in WAT, and MKP-2 was highly expressed in the stromal vascular cells (SVCs. MKP-2 inhibited the production of proinflammatory cytokines in response to FFA stimulation in macrophages. MKP-2 inhibited macrophage M1 activation through JNK and p38. In addition, overexpression of MKP-2 in macrophages suppressed inflammation during macrophage-adipocyte interaction.MKP-2 is a negative regulator of macrophage M1 activation through JNK and p38 and inhibits inflammation during macrophage-adipocyte interaction.

  12. Isolation and characterization of lipid-associated and neurosecretory polypeptides

    Stark, Margareta

    2000-01-01

    Lipid-interacting proteins play important roles in all living organisms. This thesis focuses on isolation and characterization of an enzyme in the triacylglycerol biosynthesis (phosphatidic acid phosphatase, PAP), hydrophobic polypeptides in bile, and polypeptides in cerebrospinal fluid. These fields constitute methodological challenges and mean development of suitable tools in between lipid and protein biochemistry. Two methods used to measure PAP activity were compared. I...

  13. Antimicrobial Potentials of Lactic Acid Bacteria Isolated From a Nigerian Menstruating Woman

    Funmilola Abidemi Ayeni

    2013-06-01

    Full Text Available ABSTRACT Background: Racial differences affect the composition of lactic acid bacteria (LAB in women’s vagina. However, the bacteria present in women’s vagina exert protective effect against invading uropathogens through production of several inhibitory compounds. The LAB composition of the vagina of a menstruating Nigerian woman was examined to detect any difference between the subject’s vaginal LAB flora and reported cases of women from western world and to investigate the antimicrobial activities of these lactic acid bacteria against potential uropathogens and enteropathogens with analysis of possible compounds that may be responsible for inhibition. Methods: Informed consent was obtained from the subject. LAB were identified by partially sequencing the 16S rRNA gene. The organic acids were detected through High Performance Liquid Chromatography (HPLC while the volatile compounds were detected by gas chromatography. The hydrogen peroxide production was assayed through enzymatic reactions. Results: Enterococcus faecalis FAA025 and Streptococcus equines FAA026 were the only bacterial strains isolated. The two LAB strains inhibited the growth of all tested uropathogens and enteropathogens to remarkable degree. Both strains produced high quantities of lactic acid while high quantities of hydrogen peroxide, acetic acid and ethanol were only observed in Streptococcus equines FAA026. Conclusions: The results of this study suggest that in spite of absence of lactobacilli during menstruation in the subject, other LAB present (Enterococcus faecalis FAA025 and Streptococcus equines FAA026 can exert protective effects against invading uropathogens. Also, the LAB composition of the Nigerian woman is similar to her counterparts in the West. [TAF Prev Med Bull 2013; 12(3.000: 283-290

  14. Identification and Antimicrobial Activity Detection of Lactic Acid Bacteria Isolated from Corn Stover Silage

    Li, Dongxia; Ni, Kuikui; Pang, Huili; Wang, Yanping; Cai, Yimin; Jin, Qingsheng

    2015-01-01

    A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC 43971T, Micrococcus luteus ATCC 4698T and Escherichia coli ATCC 11775T were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at 100°C), but the antimicrobial activity was eliminated after treatment at 121°C for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory. PMID:25924957

  15. A specific acid [alpha]-glucosidase in lamellar bodies of the human lung

    Vries, A.C.J. de; Schram, A.W.; Tager, J.M.; Batenburg, J.J.

    2006-01-01

    In the present investigation, we have demonstrated that three lysosomal-type hydrolases, alpha-glucosidase, alpha-mannosidase and a phosphatase, are present in lamellar bodies isolated from adult human lung. The hydrolase activities that were studied, all showed an acidic pH optimum, which is charac

  16. Identification of okadaic acid production in the marine dinoflagellate Prorocentrum rhathymum from Florida Bay

    An, Tianying; Winshell, Jamie; Scorzetti, Gloria; Fell, Jack W.; Rein, Kathleen S.

    2009-01-01

    Extracts of fifty-seven newly isolated strains of dinoflagellates and raphidophytes were screened for protein phosphatase (PP2A) inhibition. Five strains, identified by rDNA sequence analysis as Prorocentrum rhathymum, tested positive and the presence of okadaic acid was confirmed in one strain by HPLC-MS/MS and by HPLC with fluorescence detection and HPLC-MS of the okadaic acid ADAM derivative. Quantitation of the ADAM derivative indicated that the concentration of okadaic acid in the cultur...

  17. Asp1 from Schizosaccharomyces pombe binds a [2Fe-2S](2+) cluster which inhibits inositol pyrophosphate 1-phosphatase activity.

    Wang, Huanchen; Nair, Vasudha S; Holland, Ashley A; Capolicchio, Samanta; Jessen, Henning J; Johnson, Michael K; Shears, Stephen B

    2015-10-27

    Iron-sulfur (Fe-S) clusters are widely distributed protein cofactors that are vital to cellular biochemistry and the maintenance of bioenergetic homeostasis, but to our knowledge, they have never been identified in any phosphatase. Here, we describe an iron-sulfur cluster in Asp1, a dual-function kinase/phosphatase that regulates cell morphogenesis in Schizosaccharomyces pombe. Full-length Asp1, and its phosphatase domain (Asp1(371-920)), were each heterologously expressed in Escherichia coli. The phosphatase activity is exquisitely specific: it hydrolyzes the 1-diphosphate from just two members of the inositol pyrophosphate (PP-InsP) signaling family, namely, 1-InsP7 and 1,5-InsP8. We demonstrate that Asp1 does not hydrolyze either InsP6, 2-InsP7, 3-InsP7, 4-InsP7, 5-InsP7, 6-InsP7, or 3,5-InsP8. We also recorded 1-phosphatase activity in a human homologue of Asp1, hPPIP5K1, which was heterologously expressed in Drosophila S3 cells with a biotinylated N-terminal tag, and then isolated from cell lysates with avidin beads. Purified, recombinant Asp1(371-920) contained iron and acid-labile sulfide, but the stoichiometry (0.8 atoms of each per protein molecule) indicates incomplete iron-sulfur cluster assembly. We reconstituted the Fe-S cluster in vitro under anaerobic conditions, which increased the stoichiometry to approximately 2 atoms of iron and acid-labile sulfide per Asp1 molecule. The presence of a [2Fe-2S](2+) cluster in Asp1(371-920) was demonstrated by UV-visible absorption, resonance Raman spectroscopy, and electron paramagnetic resonance spectroscopy. We determined that this [2Fe-2S](2+) cluster is unlikely to participate in redox chemistry, since it rapidly degraded upon reduction by dithionite. Biochemical and mutagenic studies demonstrated that the [2Fe-2S](2+) cluster substantially inhibits the phosphatase activity of Asp1, thereby increasing its net kinase activity. PMID:26422458

  18. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic

    YAYAN SOFYAN

    2010-01-01

    Full Text Available Sarkono, Faturrahman, Sofyan Y. 2010. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic. Nusantara Bioscience 2: 38-42. The purpose of this study was to isolate, select and characterize lactic acid bacteria (LAB from abalone as a potential candidate probiotic in abalone cultivation system. Selective isolation of LAB performed using de Man Rogosa Sharpe medium. LAB isolate that potential as probiotics was screened. Selection was based on its ability to suppress the growth of pathogenic bacteria, bacterial resistance to acidic conditions and bacterial resistance to bile salts (bile. Further characterization and identification conducted to determine the species. The results showed that two of the ten isolates potential to be developed as probiotic bacteria that have the ability to inhibit several pathogenic bacteria such as Eschericia coli, Bacillus cereus dan Staphylococus aureus, able to grow at acidic condition and bile tolerance during the incubation for 24 hour. Based on the API test kit, the both of isolate identified as members of the species Lactobacillus paracasei ssp. paracasei.

  19. Metabolic activity and symbiotic interactions of lactic acid bacteria and yeasts isolated from water kefir.

    Stadie, Jasmin; Gulitz, Anna; Ehrmann, Matthias A; Vogel, Rudi F

    2013-09-01

    Water kefir is a mildly sour and alcoholic drink fermented by a stable microbial multispecies community. With its high sugar content and low amino acid concentration water kefir medium represents a demanding habitat. In this ecological niche only well adapted microorganisms which are fit to the consortium are able to grow and mutually provide essential nutrients. The synergism between main representatives of water kefir yeasts and lactobacilli was studied in a co-culture model system. Co-cultivation of yeasts and lactobacilli in water kefir medium significantly increased cell yield of all interaction partners, delineating the interaction of these water kefir isolates as mutualism. The support of Zygotorulaspora (Z.) florentina was due to the acidification of the medium by the lactobacilli, whereas lactobacilli are improved in growth by the disposal of essential nutrients produced by yeasts. The trophic interaction between Lactobacillus (Lb.) hordei and yeasts is constituted by the release of amino acids and Vitamin B6 from yeasts, whereas Lb. nagelii is supported in growth by their production of amino acids. The interaction of Z. florentina and Lb. nagelii was further examined to reveal that co-cultivation induced the yeast to release arginine, which was essential for Lb. nagelii. PMID:23664259

  20. Characterization of a retinoic acid responsive element isolated by whole genome PCR.

    Costa-Giomi, M P; Gaub, M P; Chambon, P; Abarzúa, P

    1992-01-01

    We have used whole PCR in an attempt to isolate novel retinoic acid (RA) responsive genes. We cloned several small genomic fragments from total human DNA containing putative retinoic acid responsive elements (RAREs) selected by direct binding to the retinoic acid receptor alpha (RAR alpha). We report here that an oligonucleotide containing a sequence from one of the cloned human DNA fragments, and referred to as alpha 1, functions as an authentic RARE. It is shown that both RAR alpha and RAR beta produced in Cos cells as well as in vitro translated RAR alpha bind directly and sequence-specifically to the alpha 1RARE. By mutational analysis it is demonstrated that the alpha 1RARE consists of an imperfect direct repeat of the estrogen- and thyroid hormone-related AGGTCA half-site motif separated by a 5 bp spacer. The orientation and spacing of the half-site repeats are shown to play a critical role in RAR recognition. When cloned upstream of a TK-Luc reporter, the alpha 1RARE is shown to confer responsiveness to RA in an orientation-independent fashion in F9 and CV-1 cells. The magnitude of the RA response mediated by the alpha 1RARE differed in these cell lines. Images PMID:1320257

  1. Isolation of bacterial cellulose nanocrystalline from pineapple peel waste: Optimization of acid concentration in the hydrolysis method

    Anwar, Budiman; Rosyid, Nurul Huda; Effendi, Devi Bentia; Nandiyanto, Asep Bayu Dani; Mudzakir, Ahmad; Hidayat, Topik

    2016-02-01

    Isolation of needle-shaped bacterial cellulose nanocrystalline with a diameter of 16-64 nm, a fiber length of 258-806 nm, and a degree of crystallinity of 64% from pineapple peel waste using an acid hydrolysis process was investigated. Experimental showed that selective concentration of acid played important roles in isolating the bacterial cellulose nanocrystalline from the cellulose source. To achieve the successful isolation of bacterial cellulose nanocrystalline, various acid concentrations were tested. To confirm the effect of acid concentration on the successful isolation process, the reaction conditions were fixed at a temperature of 50°C, a hydrolysis time of 30 minutes, and a bacterial cellulose-to-acid ratio of 1:50. Pineapple peel waste was used as a model for a cellulose source because to the best of our knowledge, there is no report on the use of this raw material for producing bacterial cellulose nanocrystalline. In fact, this material can be used as an alternative for ecofriendly and cost-free cellulose sources. Therefore, understanding in how to isolate bacterial cellulose nanocrystalline from pineapple peel waste has the potential for large-scale production of inexpensive cellulose nanocrystalline.

  2. Selection of probiotic lactic acid bacteria able to inhibit β-hemolytic Escherichia coli isolated from diarrheal piglets

    Malawach, T.

    2007-05-01

    Full Text Available A total of 306 isolates of lactic acid bacteria were isolated from 250 samples of piglet faeces. The strains were investigated for preliminary probiotic properties based on their stability in bile salts (0.30%and high acidity (pH 3.0. Ability to utilize protein, fat and starch, growth in the absence of vitamin B12 and growth with both aerobic and anaerobic conditions were also considered. As a result of above criteria, 20isolates were selected. Using an agar spot method, all isolates were able to inhibit β-hemolytic Escherichia coli 240/2 under aerobic and anaerobic condition. A further investigation using a co-culture techniqueshowed that only six isolates inhibited β-hemolytic E. coli 240/2, E. coli K88 and E. coli K 99 by more than 90 percent. The selected isolates were resistant to the following antibiotics: amikacin, polymyxin B andnalidixic acid; however the strains were susceptible to erythromycin and chloramphenicol. All six active isolates were identified as Lactobacillus plantarum by API 50 CH system.

  3. Isolation, modification, and aldose reductase inhibitory activity of rosmarinic acid derivatives from the roots of Salvia grandifolia.

    Kang, Jie; Tang, Yanbo; Liu, Quan; Guo, Nan; Zhang, Jian; Xiao, Zhiyan; Chen, Ruoyun; Shen, Zhufang

    2016-07-01

    To find aldose reductase inhibitors, two previously unreported compounds, grandifolias H and I, and five known compounds, including rosmarinic acid and rosmarinic acid derivatives, were isolated from the roots of Salvia grandifolia. A series of rosmarinic acid derivatives was obtained from rosmarinic acid using simple synthetic methods. The aldose reductase inhibitory activity of the isolated and synthesized compounds was assessed. Seven of the tested compounds showed moderate aldose reductase inhibition (IC50=0.06-0.30μM). The structure-activity relationship of aldose reductase inhibitory activity of rosmarinic acid derivatives was discussed for the first time. This study provided useful information that will facilitate the development of aldose reductase inhibitors. PMID:27233987

  4. Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus

    Gilbert Christophe

    2008-04-01

    Full Text Available Abstract Background Reversible phosphorylation events within a polymerisation complex have been proposed to modulate capsular polysaccharide synthesis in Streptococcus pneumoniae. Similar phosphatase and kinase genes are present in the exopolysaccharide (EPS biosynthesis loci of numerous lactic acid bacteria genomes. Results The protein sequence deduced from the wzb gene in Lactobacillus rhamnosus ATCC 9595 reveals four motifs of the polymerase and histidinol phosphatase (PHP superfamily of prokaryotic O-phosphatases. Native and modified His-tag fusion Wzb proteins were purified from Escherichia coli cultures. Extracts showed phosphatase activity towards tyrosine-containing peptides. The purified fusion protein Wzb was active on p-nitrophenyl-phosphate (pNPP, with an optimal activity in presence of bovine serum albumin (BSA 1% at pH 7.3 and a temperature of 75°C. At 50°C, residual activity decreased to 10 %. Copper ions were essential for phosphatase activity, which was significantly increased by addition of cobalt. Mutated fusion Wzb proteins exhibited reduced phosphatase activity on p-nitrophenyl-phosphate. However, one variant (C6S showed close to 20% increase in phosphatase activity. Conclusion These characteristics reveal significant differences with the manganese-dependent CpsB protein tyrosine phosphatase described for Streptococcus pneumoniae as well as with the polysaccharide-related phosphatases of Gram negative bacteria.

  5. Characterization of fatty acid modifying enzyme activity in staphylococcal mastitis isolates and other bacteria

    Lu Thea

    2012-06-01

    Full Text Available Abstract Background Fatty acid modifying enzyme (FAME has been shown to modify free fatty acids to alleviate their bactericidal effect by esterifying fatty acids to cholesterol or alcohols. Although it has been shown in previous studies that FAME is required for Staphylococcus aureus survival in skin abscesses, FAME is poorly studied compared to other virulence factors. FAME activity had also been detected in coagulase-negative staphylococci (CNS. However, FAME activity was only surveyed after a bacterial culture was grown for 24 h. Therefore if FAME activity was earlier in the growth phase, it would not have been detected by the assay and those strains would have been labeled as FAME negative. Results Fifty CNS bovine mastitis isolates and several S. aureus, Escherichia coli, and Streptococcus uberis strains were assayed for FAME activity over 24 h. FAME activity was detected in 54% of CNS and 80% S. aureus strains surveyed but none in E. coli or S. uberis. While some CNS strains produced FAME activity comparable to the lab strain of S. aureus, the pattern of FAME activity varied among strains and across species of staphylococci. All CNS that produced FAME activity also exhibited lipase activity. Lipase activity relative to colony forming units of these CNS decreased over the 24 h growth period. No relationship was observed between somatic cell count in the milk and FAME activity in CNS. Conclusions Some staphylococcal species surveyed produced FAME activity, but E. coli and S. uberis strains did not. All FAME producing CNS exhibited lipase activity which may indicate that both these enzymes work in concert to alter fatty acids in the bacterial environment.

  6. Method and Apparatus for Automated Isolation of Nucleic Acids from Small Cell Samples

    Sundaram, Shivshankar; Prabhakarpandian, Balabhaskar; Pant, Kapil; Wang, Yi

    2014-01-01

    RNA isolation is a ubiquitous need, driven by current emphasis on microarrays and miniaturization. With commercial systems requiring 100,000 to 1,000,000 cells for successful isolation, there is a growing need for a small-footprint, easy-to-use device that can harvest nucleic acids from much smaller cell samples (1,000 to 10,000 cells). The process of extraction of RNA from cell cultures is a complex, multi-step one, and requires timed, asynchronous operations with multiple reagents/buffers. An added complexity is the fragility of RNA (subject to degradation) and its reactivity to surface. A novel, microfluidics-based, integrated cartridge has been developed that can fully automate the complex process of RNA isolation (lyse, capture, and elute RNA) from small cell culture samples. On-cartridge cell lysis is achieved using either reagents or high-strength electric fields made possible by the miniaturized format. Traditionally, silica-based, porous-membrane formats have been used for RNA capture, requiring slow perfusion for effective capture. In this design, high efficiency capture/elution are achieved using a microsphere-based "microfluidized" format. Electrokinetic phenomena are harnessed to actively mix microspheres with the cell lysate and capture/elution buffer, providing important advantages in extraction efficiency, processing time, and operational flexibility. Successful RNA isolation was demonstrated using both suspension (HL-60) and adherent (BHK-21) cells. Novel features associated with this development are twofold. First, novel designs that execute needed processes with improved speed and efficiency were developed. These primarily encompass electric-field-driven lysis of cells. The configurations include electrode-containing constructs, or an "electrode-less" chip design, which is easy to fabricate and mitigates fouling at the electrode surface; and the "fluidized" extraction format based on electrokinetically assisted mixing and contacting of microbeads

  7. Technological and safety properties of lactic acid bacteria isolated from Spanish dry-cured sausages.

    Landeta, G; Curiel, J A; Carrascosa, A V; Muñoz, R; de las Rivas, B

    2013-10-01

    Technological and safety-related properties were analyzed in lactic acid bacteria isolated from Spanish dry-cured sausages in order to select them as starter cultures. In relation to technological properties, all the strains showed significative nitrate reductase activity; Lactobacillus plantarum, Lactobacillus paracasei and 52% of the Enterococcus faecium strains showed lipolytic activity and only Lactobacillus sakei strains (43%) were able to form biofilms. Related to safety aspects, E. faecium strains were the most resistant to antibiotics, whereas, L. sakei strains were the most sensitive. In relation to virulence factors, in the E. faecium strains analyzed, only the presence of efaA gene was detected. The analysis of biogenic amine production showed that most E. faecium strains and L. sakei Al-142 produced tyramine. In conclusion, L. paracasei Al-128 and L. sakei Al-143 strains possess the best properties to be selected as adequate and safe meat starter cultures. PMID:23743032

  8. Isolation and Crystal Structure of 1′,4′-Trans-diol of Abscisic Acid

    WANG Tian-Shan; ZHOU Jin-Yan; TAN Hong

    2006-01-01

    1 ′,4′-Trans-diol of abscisic acid was isolated from botrytis cinerea as a colorless crystal. The molecular and crystal structures have been determined by X-ray diffraction analysis. It crystallizes in orthorhombic system, space group P212121 with a = 6.724(3), b = 17.559(6), c =12.265(2) (A), a = β = y = 90°, V = 1448.1(8) (A)3, Z = 4, Dx = 1.222 g/cm3, F(000) = 576 and μ(MoKa) = 0.087 mm-1. The final R = 0.0628 and wR = 0.1604 for 2501 independent reflections with Rint = 0.0160 and 1679 observed reflections with I >2σ(Ⅰ). There are three intermolecular hydrogen bonds in a unit cell.

  9. In vitro effects of glycyrrhetinic acid on the growth of clinical isolates of Candida albicans.

    Pellati, Donatella; Fiore, Cristina; Armanini, Decio; Rassu, Mario; Bertoloni, Giulio

    2009-04-01

    Compounds derived from Glycyrrhiza glabra L. root have been used widely for centuries for their numerous therapeutic properties. The present study aimed to test the in vitro activity against Candida albicans strains of the compound 18-beta glycyrrhetinic acid (18-beta GA), derived from the root of Glycyrrhiza species. This antimicrobial activity was assessed using the National Committee for Clinical Laboratory Standards (NCCLS) method on C. albicans strains that were isolated from patients with recurrent vulvovaginal candidiasis (RVVC). The in vitro growth of the C. albicans strains was markedly reduced, in a pH-dependent manner, by relatively low doses (6.2 microg/mL) of 18-beta GA. The results demonstrate that 18-beta GA is a promising biological alternative for the topical treatment of recurrent vulvovaginal candidiasis (RVVC). PMID:19067381

  10. Diversity and Antimicrobial Properties of Lactic Acid Bacteria Isolated from Rhizosphere of Olive Trees and Desert Truffles of Tunisia

    Imene Fhoula; Afef Najjari; Yousra Turki; Sana Jaballah; Abdelatif Boudabous; Hadda Ouzari

    2013-01-01

    A total of 119 lactic acid bacteria (LAB) were isolated, by culture-dependant method, from rhizosphere samples of olive trees and desert truffles and evaluated for different biotechnological properties. Using the variability of the intergenic spacer 16S-23S and 16S rRNA gene sequences, the isolates were identified as the genera Lactococcus, Pediococcus, Lactobacillus, Weissella, and Enterococcus. All the strains showed proteolytic activity with variable rates 42% were EPS producers, while onl...

  11. Isolation of an acidic protein from cholesterol gallstones, which inhibits the precipitation of calcium carbonate in vitro.

    Shimizu, S.; Sabsay, B; Veis, A.; Ostrow, J D; Rege, R V; Dawes, L G

    1989-01-01

    In seeking to identify nucleating/antinucleating proteins involved in the pathogenesis of cholesterol gallstones, a major acidic protein was isolated from each of 13 samples of cholesterol gallstones. After the stones were extracted with methyl t-butyl ether to remove cholesterol, and methanol to remove bile salts and other lipids, they were demineralized with EDTA. The extracts were desalted with Sephadex-G25, and the proteins separated by PAGE. A protein was isolated, of molecular weight be...

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

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

    2008-01-01

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

  13. Acid-Tolerant Moderately Thermophilic Methanotrophs of the Class Gammaproteobacteria Isolated From Tropical Topsoil with Methane Seeps

    Islam, Tajul; Torsvik, Vigdis; Larsen, Øivind; Bodrossy, Levente; Øvreås, Lise; Birkeland, Nils-Kåre

    2016-01-01

    Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here, we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and uti...

  14. Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows’ milk cheeses

    Terzić-Vidojević Amarela

    2014-01-01

    Full Text Available The aim of this study was to identify and characterize the lactic acid bacteria (LAB of artisanal Golija raw and cooked cows’ milk cheeses traditionally manufactured without the addition of starter culture. A total of 188 Gram-positive and catalase-negative isolates of Golija cheeses were obtained from seven samples of different ripening time. Phenotypebased assays as well as rep-PCR and 16S rDNA sequence analysis were undertaken for all 188 Lstrains. The most diverse species were isolated from 20-day-old BGGO8 cheese (Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus casei/paracasei, Lactobacillus sucicola, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis bv. diacetylactis, Enterococcus faecium, Enterococcus durans and Leuconostoc mesenteroides. In other Golija cheeses Lactobacillus reuteri, Lactobacillus curvatus, Lactobacillus rhamnosus, Lactococcus lactis subsp. cremoris, Lactococcus garvieae, Streptococcus thermophilus and Leuconostoc pseudomesenteroides were found. Pronounced antimicrobial properties showed enterococci (13/42 and lactococci (12/31, while the good proteolytic activity demonstrated lactococci (13/31 and lactobacilli (10/29. [Projekat Ministarstva nauke Republike Srbije, br. 173019

  15. Lactic acid bacteria from "Sheep's Dhan", a traditional butter: Isolation, identification and major technological traits

    2009-06-01

    Full Text Available Twenty six lactic acid bacteria were isolated from sheep’s Dhan, a traditional butter made from sheep’s milk in Jijel (East of Algeria. These strains belong to three genera: Lactococcus, Leuconostoc and Lactobacillus. The results showed that Lactococcus lactis ssp diacetylactis was the predominant species in this traditional butter. The results of the assessment of the technological aptitude indicate that a major strain has a good acidification aptitude, some of them show good proteolytic activity and only Leuconostoc mesenteroides ssp. dextranicum isolates were able to produce exopolysaccharide.

    Veintiséis bacterias lácticas fueron aisladas de “Sheep´s Dhan”, una mantequilla tradicional hecha con leche de oveja en Jijel (al Este de Argelia. Estas cepas pertenecen a tres géneros: Lactococcus, Leuconostoc y Lactobacillus. Los resultados mostraron que Lactococcus lactis ssp diacetylactis fue la especie predominante en esta mantequilla tradicional. Los resultados de la evaluación de la aptitud tecnológica indican que la principal cepa tiene una buena aptitud de acidificación, algunas de ellas mostraron una buena actividad proteolítica y únicamente Leuconostoc mesenteroides ssp. dextranicum fue capaz de producir exopolisacárido.

  16. Isolation, Identification and Characterization of Two Aluminum-Tolerant Fungi from Acidic Red Soil.

    He, Genhe; Wang, Xiaodong; Liao, Genhong; Huang, Shoucheng; Wu, Jichun

    2016-09-01

    Acidic red soil from a forest in Jiangxi Province was selected to isolate aluminum (Al)-resistant microbes, from which eight fungi were isolated. Two strains (S4 and S7) were found to be extremely tolerant to Al concentrations of up to 550 mmol L(-1) and could grow at low pH levels (3.20-3.11). Morphological and 26S rDNA sequence analyses indicated that strain S4 belonged to Eupenicillium, while strain S7 was an unclassified Trichocomaceae. Further investigation showed that both strains were endowed with the ability to resist Al; strain S4 accumulated such a substantial amount of Al that its growth was limited to a larger extent than strain S7. The lower amounts of Al adsorbed in the mycelium and the much larger amounts of Al retained in the medium, in addition to the color change of the culture solution, implied that these two strains may resist Al by preventing Al from entering the cell and by chelating Al by secreting unique metabolites outside of the cell. PMID:27407299

  17. Isolation of Endotoxin Eliminating Lactic Acid Bacteria and a Property of Endotoxin Eliminating Protein.

    Kondo, Ayaka; Asami, Kyoko; Suda, Yoshihito; Shimoyamada, Makoto; Kanauchi, Makoto

    2016-06-01

    Recently, many scholars have reported lactic acid bacteria (LAB) functions, such as anticancer activity and anti-inflammatory activity for intestines. To decrease inflammatory substances such as endotoxins, LAB consumed safely with meals were isolated from food and food ingredients. First, LAB were isolated as 168 strains of bacillus LAB (49 strain) and coccus LAB (119 strains) from food ingredients and fermented foods such as rice, rice bran, malt, grains, miso soy paste, and some pickles. Their LAB (168 strains) were cultivated in medium containing endotoxin from Escherichia coli O18 LPS at 15 and 30 °C for 64 h to identify endotoxin-eliminating LAB. Consequently, the AK-23 strain was screened as an endotoxin-eliminating LAB strain. The strain decreased endotoxin in YP medium without sugar at 30 °C for 64 h until 9% of endotoxin. The strain was identified as Pediococcus pentosaceus according to morphological characteristics such as its cell shape, physiological characteristics related to its fermentation type, assimilation of sugars, pH tolerance, optimum growth temperature, and molecular biological characteristics as its homology to 16S rRNA. To investigate the location of the endotoxin-eliminating substance, 4 fractions were separated from AK-23 cells as extracellular, cell wall digestion, cytoplasm, and cell membrane fractions. The endotoxin-decreasing substance, located on a cell wall, was identified as a 217 kDa protein. PMID:27096744

  18. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Ghabbour, N.

    2011-03-01

    Full Text Available A total of 177 strains of lactic acid bacteria (LAB were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%, Lactobacillus pentosus (25.99%, Lactobacillus brevis (9.61% and Pediococcus pentosaceus (19.77%. All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3% were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo- 4-chloro-3-indolyl β-D-glucuronide (X-Gluc as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis.

    Un total de 177 cepas de bacterias ácido lácticas (LAB fueron aisladas en las primeras etapas de la fermentación de aceitunas verdes marroquíes Picholine, incluyendo Lactobacillus plantarum (44.63%, Lactobacillus pentosus (25.99%, Lactobacillus brevis (9.61% y Pediococcus pentosaceus (19.77%. Todos los aislados fueron evaluados mediante su tolerancia a extractos de hojas de olivo y oleuropeína. La mayoría de los aislados (85,3% degradaron oleuropeína, cuando fueron evaluados usando oleuropeína o 5-Bromo-4-cloro- 3-indolil β-D-glucuronido (X-Gluc como sustrato. La capacidad de biodegradación de las cepas seleccionadas para cada especie fue confirmada mediante análisis por HPLC.

  19. Isolation of actinides and lanthanides by flotation from nitric acid solutions in the form of the complexes with diphosphinedioxides

    Isolation of europium, plutonium (4) and americium by ionic flotation from nitric acid solutions (1-5 mol/l HNO3), using diphosphinedioxides as surfactant-precipitator, was investigated. Increase in the metal isolation degree with an increase in flotation agent consumption was ascertained. When flotation agent is in excess, required for precipitation completeness, the metals studied are noticeably extracted from moderately concentrated solutions of nitric acid (3-3.5 mol/l), moreover, tolyl-containing diphosphinedioxide per one flotation operation extracts ∼90 % of plutonium, ∼75 % of americium and >75 % of europium

  20. Extreme Elevation of Alkaline Phosphatase in a Pregnancy Complicated by Gestational Diabetes and Infant with Neonatal Alloimmune Thrombocytopenia.

    Lozo, Svjetlana; Atabeygi, Amir; Healey, Michael

    2016-01-01

    There have been few case reports of isolated elevation of alkaline phosphatase beyond the normal physiologic amount with subsequent return to baseline after delivery. Here we present a similar case of extreme elevation of alkaline phosphatase in a pregnancy complicated by gestational diabetes and subsequently by neonatal alloimmune thrombocytopenia (NAIT). PMID:27610256

  1. Toward Probiotict Food Product from Meat Through Isolation and Identification Lactic Acid Bacteria As Probiotic Culture Stater

    Yunilas Yunilas

    2014-01-01

    Full Text Available Probiotic food products of meat can provide extensive benefits, to increase the shelf life and nutritional value also improve the taste. The use of lactic acid bacteria culture (LAB derived from the isolation of the meat and the addition of probiotic cultures (Lactobacilli and Bifidobacteria in fermented sausage processing is expected to produce a probiotic sausage products with nutritional value, and better shelf life and improve health. This study aimed to isolate and identify lactic acid bacteria (LAB of meat as a starter culture fermented sausages. The parameters observed were gram test, catalase, motility, gas production, type of fermentation, growth at various temperatures and pH. The results were obtained 28 isolates, 17 isolates were able to produce acid and 8 of them are lactic acid bacteria (LAB with the characteristics of gram-positive, catalase negative, not motile, produces gas, are hetero and homo fermentative, optimum growth temperature of 300C and a few of them are able to grow on pH 3.5. Lactic acid bacteria that able to be combined with probiotics as sausage starter culture to the probiotic food products of meat.

  2. Desulfurella amilsii sp. nov., a novel acidotolerant sulfur-respiring bacterium isolated from acidic river sediments.

    Florentino, Anna P; Brienza, Claudio; Stams, Alfons J M; Sánchez-Andrea, Irene

    2016-03-01

    A novel acidotolerant and moderately thermophilic sulfur-reducing bacterium was isolated from sediments of the Tinto River (Spain), an extremely acidic environment. Strain TR1T stained Gram-negative, and was obligately anaerobic, non-spore-forming and motile. Cells were short rods (1.5-2 × 0.5-0.7 μm), appearing singly or in pairs. Strain TR1T was catalase-negative and slightly oxidase-positive. Urease activity and indole formation were absent, but gelatin hydrolysis was present. Growth was observed at 20-52 °C with an optimum close to 50 °C, and a pH range of 3-7 with optimum between pH 6 and 6.5. Yeast extract was essential for growth, but extra vitamins were not required. In the presence of sulfur, strain TR1T grew with acetate, formate, lactate, pyruvate, stearate, arginine and H2/CO2. All substrates were completely oxidized and H2S and CO2 were the only metabolic products detected. Besides elemental sulfur, thiosulfate was used as an electron acceptor. The isolate also grew by disproportionation of elemental sulfur. The predominant cellular fatty acids were saturated components: C16 : 0, anteiso-C17 : 0 and C18 : 0. The only quinone component detected was menaquinone MK-7(H2). The G+C content of the genomic DNA was 34 mol%. The isolate is affiliated to the genus Desulfurella of the class Deltaproteobacteria, sharing 97 % 16S rRNA gene sequence similarity with the four species described in the genus Desulfurella. Considering the distinct physiological and phylogenetic characteristics, strain TR1T represents a novel species within the genus Desulfurella, for which the name Desulfurella amilsii sp. nov. is proposed. The type strain is TR1T ( = DSM 29984T = JCM 30680T). PMID:26704766

  3. Trends for isolated amino acids and dipeptides: Conformation, divalent ion binding, and remarkable similarity of binding to calcium and lead

    Ropo, Matti; Baldauf, Carsten

    2016-01-01

    We derive structural and binding energy trends for twenty amino acids, their dipeptides, and their interactions with the divalent cations Ca$^{2+}$, Ba$^{2+}$, Sr$^{2+}$, Cd$^{2+}$, Pb$^{2+}$, and Hg$^{2+}$. The underlying data set consists of 45,892 first-principles predicted conformers with relative energies up to about 4 eV (about 400kJ/mol). We show that only very few distinct backbone structures of isolated amino acids and their dipeptides emerge as lowest-energy conformers. The isolated amino acids predominantly adopt structures that involve an acidic proton shared between the carboxy and amino function. Dipeptides adopt one of two intramolecular-hydrogen bonded conformations C$_5$ or equatorial C$_7$. Upon complexation with a divalent cation, the accessible conformational space shrinks and intramolecular hydrogen bonding is prevented due to strong electrostatic interaction of backbone and side chain functional groups with cations. Clear correlations emerge from the binding energies of the six divalent ...

  4. Pantethine inhibits cholesterol and fatty acid syntheses and stimulates carbon dioxide formation in isolated rat hepatocytes.

    Cighetti, G; Del Puppo, M; Paroni, R; Fiorica, E; Galli Kienle, M

    1987-02-01

    The effects of pantethine on cholesterol and fatty acid metabolism were investigated in isolated rat hepatocytes. Preincubation of the cells with pantethine induced a concentration-dependent decrease of the radioactivity incorporated into carbon dioxide and lipids in incubations with [2-14C]acetate. When pantethine and the labeled substrate were simultaneously added to the cell suspension, there was an enhancement of carbon dioxide radioactivity at short incubation time (5 min) whereas, at longer incubation time, values were comparable to those of controls; lipid radioactivity, instead, was dramatically reduced by pantethine even at short incubation time and decreased further during the incubation, being 23% of that of controls at 60 min. Analysis of the incubation medium showed that pantethine induced a concentration- and time-dependent release of acetate into the medium. Results of the effect of the acetate concentration on the incorporation of [2-14C]acetate radioactivity into CO2 and lipids in control hepatocytes allowed the conclusion that the above-described modifications induced by pantethine are only partially attributable to the dilution of the labeled substrate, and that catabolism of acetate to carbon dioxide is stimulated by the disulphide pantethine, whereas cholesterol and fatty acid syntheses are inhibited. PMID:3106549

  5. Reduction of Ammonia Loss from Urea through Mixing with Humic Acids Isolated from Peat Soil (Saprists

    Regis Bernard

    2009-01-01

    Full Text Available Problem statement: Application of urea as a source of nitrogen fertilizer has an adverse effect on ammoniacal loss to the environment. This study was conducted to reduce ammonia loss from urea by mixing with Humic Acids (HA isolated from Saprists peat. Approach: The effects of urea amended with four different amounts of humic acids, 0.25, 0.50, 0.75 and 1.00 g were evaluated in laboratory conditions using a closed dynamic air flow system. The mineral soil that was used as medium for the study was Bekenu series (typic paleudults. Amnonia loss, soil pH, exchangeable ammonium, available nitrate, exchangeable K, Ca, Mg and Na were determined using standard procedures. Results: All the treatments with HA significantly reduced ammoinia loss compared to urea alone. Increasing the amount of HA also significantly retained soil exchangeable ammonium and available nitrate. Treatments with HA had no significant effect on the concentrations of Mg, K and Ca, except for Na. The effect of HA in the mixtures on ammonia loss was related to their effect on the formation of ammonium over ammonia. Conclusion: Surface-applied urea fertilizer efficiency could be increased when coated with 1.00 g of HA.

  6. Fatty acid synthase inhibitors from plants: isolation, structure elucidation, and SAR studies.

    Li, Xing-Cong; Joshi, Alpana S; ElSohly, Hala N; Khan, Shabana I; Jacob, Melissa R; Zhang, Zhizheng; Khan, Ikhlas A; Ferreira, Daneel; Walker, Larry A; Broedel, Sheldon E; Raulli, Robert E; Cihlar, Ronald L

    2002-12-01

    Fatty acid synthase (FAS) has been identified as a potential antifungal target. FAS prepared from Saccharomyces cerevisiae was employed for bioactivity-guided fractionation of Chlorophora tinctoria,Paspalum conjugatum, Symphonia globulifera, Buchenavia parviflora, and Miconia pilgeriana. Thirteen compounds (1-13), including three new natural products (1, 4, 12), were isolated and their structures identified by spectroscopic interpretation. They represented five chemotypes, namely, isoflavones, flavones, biflavonoids, hydrolyzable tannin-related derivatives, and triterpenoids. 3'-Formylgenistein (1) and ellagic acid 4-O-alpha-l-rhamnopyranoside (9) were the most potent compounds against FAS, with IC(50) values of 2.3 and 7.5 microg/mL, respectively. Furthermore, 43 (14-56) analogues of the five chemotypes from our natural product repository and commercial sources were tested for their FAS inhibitory activity. Structure-activity relationships for some chemotypes were investigated. All these compounds were further evaluated for antifungal activity against Candida albicans and Cryptococcus neoformans. Although there were several antifungal compounds in the set, correlation between the FAS inhibitory activity and antifungal activity could not be defined. PMID:12502337

  7. A mutation in protein phosphatase 2A regulatory subunit A affects auxin transport in Arabidopsis

    Garbers, C.; DeLong, A.; Deruere, J.; Bernasconi, P.; Soll, D.; Evans, M. L. (Principal Investigator)

    1996-01-01

    The phytohormone auxin controls processes such as cell elongation, root hair development and root branching. Tropisms, growth curvatures triggered by gravity, light and touch, are also auxin-mediated responses. Auxin is synthesized in the shoot apex and transported through the stem, but the molecular mechanism of auxin transport is not well understood. Naphthylphthalamic acid (NPA) and other inhibitors of auxin transport block tropic curvature responses and inhibit root and shoot elongation. We have isolated a novel Arabidopsis thaliana mutant designated roots curl in NPA (rcn1). Mutant seedlings exhibit altered responses to NPA in root curling and hypocotyl elongation. Auxin efflux in mutant seedlings displays increased sensitivity to NPA. The rcn1 mutation was transferred-DNA (T-DNA) tagged and sequences flanking the T-DNA insert were cloned. Analysis of the RCN1 cDNA reveals that the T-DNA insertion disrupts a gene for the regulatory A subunit of protein phosphatase 2A (PP2A-A). The RCN1 gene rescues the rcn1 mutant phenotype and also complements the temperature-sensitive phenotype of the Saccharomyces cerevisiae PP2A-A mutation, tpd3-1. These data implicate protein phosphatase 2A in the regulation of auxin transport in Arabidopsis.

  8. Protein tyrosine phosphatases expression during development of mouse superior colliculus

    Reinhard, Jacqueline; Horvat-Bröcker, Andrea; Illes, Sebastian; Zaremba, Angelika; Knyazev, Piotr; Ullrich, Axel; Faissner, Andreas

    2009-01-01

    Protein tyrosine phosphatases (PTPs) are key regulators of different processes during development of the central nervous system. However, expression patterns and potential roles of PTPs in the developing superior colliculus remain poorly investigated. In this study, a degenerate primer-based reverse transcription-polymerase chain reaction (RT-PCR) approach was used to isolate seven different intracellular PTPs and nine different receptor-type PTPs (RPTPs) from embryonic E15 mouse superior col...

  9. [Study of pantothenic acid derivatives as cardiac protectors in a model of experimental ischemia and reperfusion of the isolated heart].

    Kumerova, A O; Utno, L Ia; Lipsberga, Z E; Shkestere, I Ia

    1992-04-01

    An isolated heart model with experimental ischemia and reperfusion was used to show effective decrease in lactate, increase in ATP content and prevention of conjugated dienes accumulation in the myocardium by derivatives of pantothenic acid: panthenol (9.0 mg/kg), calcium pantothenate (15.6 mg/kg) and by these ones applied simultaneously as ingredients of perfusate (25 microM) in postischemic period. In that way derivatives of pantothenic acid should be regarded as cardiac protectors. PMID:1391892

  10. 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

    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.

  11. Asiatic acid uncouples respiration in isolated mouse liver mitochondria and induces HepG2 cells death.

    Lu, Yapeng; Liu, Siyuan; Wang, Ying; Wang, Dang; Gao, Jing; Zhu, Li

    2016-09-01

    Asiatic acid, one of the triterpenoid components isolated from Centella asiatica, has received increasing attention due to a wide variety of biological activities. To date, little is known about its mechanisms of action. Here we examined the cytotoxic effect of asiatic acid on HepG2 cells and elucidated some of the underlying mechanisms. Asiatic acid induced rapid cell death, as well as mitochondrial membrane potential (MMP) dissipation, ATP depletion and cytochrome c release from mitochondria to the cytosol in HepG2 cells. In mitochondria isolated from mouse liver, asiatic acid treatment significantly stimulated the succinate-supported state 4 respiration rate, dissipated the MMP, increased Ca(2+) release from Ca(2+)-loaded mitochondria, decreased ATP content and promoted cytochrome c release, indicating the uncoupling effect of asiatic acid. Hydrogen peroxide (H2O2) produced by succinate-supported mitochondrial respiration was also significantly inhibited by asiatic acid. In addition, asiatic acid inhibited Ca(2+)-induced mitochondrial swelling but did not induce mitochondrial swelling in hyposmotic potassium acetate medium which suggested that asiatic acid may not act as a protonophoric uncoupler. Inhibition of uncoupling proteins (UCPs) or blockade of adenine nucleotide transporter (ANT) attenuated the effect of asiatic acid on MMP dissipation, Ca(2+) release, mitochondrial respiration and HepG2 cell death. When combined inhibition of UCPs and ANT, asiatic acid-mediated uncoupling effect was noticeably alleviated. These results suggested that both UCPs and ANT partially contribute to the uncoupling properties of asiatic acid. In conclusion, asiatic acid is a novel mitochondrial uncoupler and this property is potentially involved in its toxicity on HepG2 cells. PMID:27288117

  12. Fatty Acid Profiling of Lipid A Isolated from Indigenous salmonella typhi strain by gas chromatography mass spectrometry

    Typhoid, caused by Salmonella enterica serovar Typhi (S. Typhi), is a major health problem worldwide especially in developing countries. Lipopolysaccharides are one of the main virulence factors of S. Typhi. Hydrophobic lipid A anchors the lipopolysaccharides into the bacterial outer membrane and also serves as the epicenter of endotoxicity, which is linked to the presence of several fatty acid chains. Fatty acid profiling is, therefore, very important to understand the endotoxicity of these pathogenic bacteria. To profile lipid A with respect to its fatty acid constituents, a S. Typhi was isolated from blood culture of a typhoid patient from the Faisalabad region of Pakistan. After its complete identification using biochemical and molecular techniques, this bacterium was cultivated in a fermentor. The cell pellet obtained was subjected to hot phenol process to extract and purify lipopolysaccharides. Acid hydrolysis of the lipopolysaccharides yielded lipid A, which was subjected to analyses using GC-MS after derivatization into their fatty acid methyl esters. The fatty acid methyl esters were identified on the basis of their retention times, compared with standards as well as characteristic mass fragmentation patterns of their respective mass spectra. This fatty acid profiling revealed the occurrence of dodecanoic acid (C12:0), tetradecanoic acid (C14:0), 3-hydroxy tetradecanoic acid (3-OH C14:0) and hexadecanoic acid (C16:0) in the lipid A component of S. Typhi strain with the relative percentage abundances 8.5%, 12.5%, 55.9% and 23.1%, respectively. (author)

  13. Catalytic DNA with phosphatase activity

    Chandrasekar, Jagadeeswaran; Silverman, Scott K.

    2013-01-01

    Catalytic DNA sequences (deoxyribozymes, DNA enzymes, or DNAzymes) have been identified by in vitro selection for various catalytic activities. Expanding the limits of DNA catalysis is an important fundamental objective and may facilitate practical utility of catalysts that can be obtained from entirely unbiased (random) sequence populations. In this study, we show that DNA can catalyze Zn2+-dependent phosphomonoester hydrolysis of tyrosine and serine side chains (i.e., exhibit phosphatase ac...

  14. Structure of a novel multidrug resistance modulator, irciniasulfonic acid, isolated from a marine sponge, Ircinia sp.

    A. Kawakami; T. Miyamoto; R. Higuchi; T. Uchiumi; M. Kuwano; R.W.M. van Soest

    2001-01-01

    Irciniasulfonic acid was obtained from a marine sponge of Ircinia sp. Spectroscopic and chemical analyses revealed its structure consists of three different kinds of acids, i.e. common fatty acids, a novel unsaturated branched C-10 fatty acid and an isethionic acid. Irciniasulfonic acid and deacyl i

  15. Control of placental alkaline phosphatase gene expression in HeLa cells: induction of synthesis by prednisolone and sodium butyrate

    HeLa S3 cells produce an alkaline phosphatase indistinguishable from the enzyme from human term placenta. The phosphatase activity in these cells was induced by both prednisolone and sodium butyrate. Both agents stimulated de novo synthesis of the enzyme. The increase in phosphatase activity paralleled the increase in immunoactivity and biosynthesis of placental alkaline phosphatase. The fully processed phosphatase monomer in control, prednisolone-treated or butyrate-treated cells was a 64.5 K polypeptide, measured by both incorporation of L-[35S]methionine into enzyme protein and active-site labeling. The 64.5K polypeptide was formed by the incorporation of additional N-acetylneuraminic acid moieties to a precursor polypeptide of 61.5K. However, this biosynthetic pathway was identified only in butyrate-treated cells. In prednisolone-treated cells, the processing of 61.5K to 64.5K monomer was accelerated, and the presence of the 61.5 precursor could only be detected by either neuraminidase or monensin treatment. Phosphatase mRNA which comigrated with the term placental alkaline phosphatase mRNA of 2.7 kilobases was induced in the presence of either prednisolone or butyrate. Alkaline phosphatase mRNA is untreated HeLa S3 cells migrated slightly faster than the term placental alkaline phosphatase mRNA. Butyrate also induced a second still faster migrating alkaline phosphatase mRNA. Both prednisolone and butyrate increased the steady-state levels of placental alkaline phosphatase mRNA. The data indicate that the increase in phosphatase mRNA by prednisolone and butyrate resulted in the induction of alkaline phosphatase activity and biosynthesis in HeLa S3 cells. Furthermore, both agents induced the expression of different alkaline phosphatase gene transcripts without altering its protein product

  16. Acid-tolerant moderately thermophilic methanotrophs of the class Gammaproteobacteria isolated from tropical topsoil with methane seeps

    Tajul eIslam

    2016-06-01

    Full Text Available Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and utilized methane and methanol as sole carbon and energy source. Isolates grew at pH range 4.2–7.5 (optimal 5.5–6.0 and at a temperature range of 30–60oC (optimal 51–55oC. 16S rRNA gene phylogeny placed them in a well-separated branch forming a cluster together with the genus Methylocaldum as the closest relatives (93.1–94.1% sequence similarity. The genes pmoA, mxaF, and cbbL were detected, but mmoX was absent. Strains BFH1 and BFH2 are, to our knowledge, the first isolated acid-tolerant moderately thermophilic methane oxidizers of the class Gammaproteobacteria. Each strain probably denotes a novel species and they most likely represent a novel genus within the family Methylococcaceae of type I methanotrophs. Furthermore, the isolates increase our knowledge of acid-tolerant aerobic methanotrophs and signify a previously unrecognized biological methane sink in tropical ecosystems.

  17. Acid-Tolerant Moderately Thermophilic Methanotrophs of the Class Gammaproteobacteria Isolated From Tropical Topsoil with Methane Seeps

    Islam, Tajul; Torsvik, Vigdis; Larsen, Øivind; Bodrossy, Levente; Øvreås, Lise; Birkeland, Nils-Kåre

    2016-01-01

    Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here, we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and utilized methane and methanol as sole carbon and energy source. Isolates grew at pH range 4.2–7.5 (optimal 5.5–6.0) and at a temperature range of 30–60°C (optimal 51–55°C). 16S rRNA gene phylogeny placed them in a well-separated branch forming a cluster together with the genus Methylocaldum as the closest relatives (93.1–94.1% sequence similarity). The genes pmoA, mxaF, and cbbL were detected, but mmoX was absent. Strains BFH1 and BFH2 are, to our knowledge, the first isolated acid-tolerant moderately thermophilic methane oxidizers of the class Gammaproteobacteria. Each strain probably denotes a novel species and they most likely represent a novel genus within the family Methylococcaceae of type I methanotrophs. Furthermore, the isolates increase our knowledge of acid-tolerant aerobic methanotrophs and signify a previously unrecognized biological methane sink in tropical ecosystems. PMID:27379029

  18. Acid-Tolerant Moderately Thermophilic Methanotrophs of the Class Gammaproteobacteria Isolated From Tropical Topsoil with Methane Seeps.

    Islam, Tajul; Torsvik, Vigdis; Larsen, Øivind; Bodrossy, Levente; Øvreås, Lise; Birkeland, Nils-Kåre

    2016-01-01

    Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here, we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and utilized methane and methanol as sole carbon and energy source. Isolates grew at pH range 4.2-7.5 (optimal 5.5-6.0) and at a temperature range of 30-60°C (optimal 51-55°C). 16S rRNA gene phylogeny placed them in a well-separated branch forming a cluster together with the genus Methylocaldum as the closest relatives (93.1-94.1% sequence similarity). The genes pmoA, mxaF, and cbbL were detected, but mmoX was absent. Strains BFH1 and BFH2 are, to our knowledge, the first isolated acid-tolerant moderately thermophilic methane oxidizers of the class Gammaproteobacteria. Each strain probably denotes a novel species and they most likely represent a novel genus within the family Methylococcaceae of type I methanotrophs. Furthermore, the isolates increase our knowledge of acid-tolerant aerobic methanotrophs and signify a previously unrecognized biological methane sink in tropical ecosystems. PMID:27379029

  19. Estudio de la fosfatasa ácida y alcalina en suelos de la Región Pampeana Norte del área sojera argentina Study of acid and alkaline phosphatase in soils of the Pampean North Region from argentine soybean area

    Leticia Andrea Fernández

    2008-07-01

    ón de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po.Transformation of organic phosphorus (Po into soluble inorganic phosphorus (Pi is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB and cultivated fungi (CF was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5 CFU g-1 soil, while alkaline activity was 5.80 10(5 CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5 mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

  20. Emerging nalidixic acid and ciprofloxacin resistance in non-typhoidal Salmonella isolated from patients having acute diarrhoeal disease

    Non-typhoidal Salmonella are one of the key etiological agents of diarrhoeal disease. The appearence of multiple drung resistance along with resistance to quinolones in this bacterium poses a serious therapeutic problem. We determined the prevalence of nalidixic acid and ciprofloxacin resistance in non-typhodial Salmonella isolated from faecal samples of patients with acute diarroheal disease attending the outpatient and inpatient department of a hospital in Saudi Arabia during the years 1999 to 2002. Non-typhodial Salmonella were isolated from faecal samples. Antimicrobial susceptibility was tested by the disc diffusion test. MICs to nalidixic acid and ciprofloxacinwere determined by the agar dilution method. During the study period , 524 strains of non-typhoidal Salmonella were isolated. Strains belonging to serogroup C1were the commonest (41.4%) followed by serogroups B and D (15.6% and 14.5%, respectively). Resistance to ampicillin was observed in 22.9% and to trimethoprim/sulphamethoxazole in 18.5%of the strains. Nalidixic acid resistance was encounterd in 9.9% and ciprofloxacin esistance in 2.3% of the strains. Resistance to nalidixic acid significantly increased from 0.1% in 1999 to 5.51% in 2002 ( p=0.0007)and ciprofloxacin resistance increased significantly from 0.1% in 1999 to 0.9% in 2002( p=0.0001). MICs to nalidixic acid and ciprofloxacin were determined among 29 nalidixic acid-resistant strains of non-typhoidal salmonella isolated during 2002. The MIC was >256 ug /ml to nalidixic acid and 8 to 16 ug/ml to ciprofloxacin. The increasing rate of antimicrobial resistance encountered among non-tyophoidal Salmonella necessiate the judicious use of these drugs in humans. Moreover, these findings support the concern that the use of quinolones in animal feed may lead to an increasein resistance and should should be restricted. (author)

  1. Isolation from swine feces of a bacterium which decarboxylates p-hydroxyphenylacetic acid to 4-methylphenol (p-cresol).

    L. A. Ward; Johnson, K A; Robinson, I.M.; Yokoyama, M T

    1987-01-01

    An obligate anaerobe has been isolated from swine feces which decarboxylates p-hydroxyphenylacetic acid to 4-methylphenol (p-cresol). The bacterium was an ovoid rod, gram positive, nonsporeforming, and nonmotile. Lactate and acetate were major end products of glucose fermentation. Based on its characteristics, the bacterium is tentatively assigned to the genus Lactobacillus.

  2. Determination of Minimal Duration Essential for Isolation of Humic Acids From Soils in Forest Restoration Programmes

    Mohd R. N. Hanisah

    2008-01-01

    Full Text Available This study was conducted to investigate whether a simple and rapid method could be developed for extracting, fractionating and purifying soil HA in forest rehabilitation programmes. Humic acids from 10 g of soil were extracted with 100 mL of 0.10 M NaOH. Different extraction periods (4, 8, 12, 16, 20 and 24 h were tested. Samples were centrifuged (16,211 G for 15 min at the end of each extraction period. The dark-coloured supernatant liquor containing HA was decanted and the pH of the solution adjusted to 1.0 using 6 M HCl. After acidification, the fractionation periods evaluated were 4, 8, 12, 16, 20 and 24 h. After each fractionation period, the sample was transferred to a polyethylene bottle and centrifuged (16,211 G for 10 min. The HA were purified by suspending them in 100 mL distilled water, centrifuged (16,211 G for 10 min. After repeating this procedure three times, the supernatant was analyzed for Na, Mg and K. Standard procedures were used to characterize the HA (C, E4/E6, phenolic OH, carboxylic COOH, total acidity and soil (pH, C, organic matter. Although there was significant effect of different extraction periods on yield of HA, there was no significant relationship between fractionation period and yield of HA. There was also no significant relationship between fractionation periods and yield of HA for different extraction periods studied. In terms of purification, the distilled water used in this study was able to effectively purify HA (e.g., reduction in mineral matter such as Na+ of the soil without altering the true nature of HA as C, E4/E6, phenolic OH, carboxylic COOH, total acidity values of the acids were consistent with those reported in the literature. The significance of this work is that it enables the isolation of HA from soil within 9 h (4 h extraction period, 4 h fractionation period and 1 h purification period instead of the existing range of 2-7 days, hence helping in facilitating the idea of producing for

  3. Polyploid genome of Camelina sativa revealed by isolation of fatty acid synthesis genes

    Shewmaker Christine K

    2010-10-01

    Full Text Available Abstract Background Camelina sativa, an oilseed crop in the Brassicaceae family, has inspired renewed interest due to its potential for biofuels applications. Little is understood of the nature of the C. sativa genome, however. A study was undertaken to characterize two genes in the fatty acid biosynthesis pathway, fatty acid desaturase (FAD 2 and fatty acid elongase (FAE 1, which revealed unexpected complexity in the C. sativa genome. Results In C. sativa, Southern analysis indicates the presence of three copies of both FAD2 and FAE1 as well as LFY, a known single copy gene in other species. All three copies of both CsFAD2 and CsFAE1 are expressed in developing seeds, and sequence alignments show that previously described conserved sites are present, suggesting that all three copies of both genes could be functional. The regions downstream of CsFAD2 and upstream of CsFAE1 demonstrate co-linearity with the Arabidopsis genome. In addition, three expressed haplotypes were observed for six predicted single-copy genes in 454 sequencing analysis and results from flow cytometry indicate that the DNA content of C. sativa is approximately three-fold that of diploid Camelina relatives. Phylogenetic analyses further support a history of duplication and indicate that C. sativa and C. microcarpa might share a parental genome. Conclusions There is compelling evidence for triplication of the C. sativa genome, including a larger chromosome number and three-fold larger measured genome size than other Camelina relatives, three isolated copies of FAD2, FAE1, and the KCS17-FAE1 intergenic region, and three expressed haplotypes observed for six predicted single-copy genes. Based on these results, we propose that C. sativa be considered an allohexaploid. The characterization of fatty acid synthesis pathway genes will allow for the future manipulation of oil composition of this emerging biofuel crop; however, targeted manipulations of oil composition and general

  4. Rhizobium acidisoli sp. nov., isolated from root nodules of Phaseolus vulgaris in acid soils.

    Román-Ponce, Brenda; Jing Zhang, Yu; Soledad Vásquez-Murrieta, María; Hua Sui, Xin; Feng Chen, Wen; Carlos Alberto Padilla, Juan; Wu Guo, Xian; Lian Gao, Jun; Yan, Jun; Hong Wei, Ge; Tao Wang, En

    2016-01-01

    Two Gram-negative, aerobic, non-motile, rod-shaped bacterial strains, FH13T and FH23, representing a novel group of Rhizobium isolated from root nodules of Phaseolus vulgaris in Mexico, were studied by a polyphasic analysis. Phylogeny of 16S rRNA gene sequences revealed them to be members of the genus Rhizobium related most closely to 'Rhizobium anhuiense' CCBAU 23252 (99.7 % similarity), Rhizobium leguminosarum USDA 2370T (98.6 %), and Rhizobium sophorae CCBAU 03386T and others ( ≤ 98.3 %). In sequence analyses of the housekeeping genes recA, glnII and atpD, both strains formed a subclade distinct from all defined species of the genus Rhizobium at sequence similarities of 82.3-94.0 %, demonstrating that they represented a novel genomic species in the genus Rhizobium. Mean levels of DNA-DNA relatedness between the reference strain FH13T and the type strains of related species varied between 13.0 ± 2.0 and 52.1 ± 1.2 %. The DNA G+C content of strain FH13T was 63.5 mol% (Tm). The major cellular fatty acids were 16 : 0, 17 : 0 anteiso, 18 : 0, summed feature 2 (12 : 0 aldehyde/unknown 10.928) and summed feature 8 (18 : 1ω7c). The fatty acid 17 : 1ω5c was unique for this strain. Some phenotypic features, such as failure to utilize adonitol, l-arabinose, d-fructose and d-fucose, and ability to utilize d-galacturonic acid and itaconic acid as carbon source, could also be used to distinguish strain FH13T from the type strains of related species. Based upon these results, a novel species, Rhizobium acidisoli sp. nov., is proposed, with FH13T ( = CCBAU 101094T = HAMBI 3626T = LMG 28672T) as the type strain. PMID:26530784

  5. Antimicrobial Activity and Antibiotic Sensitivity of Three Isolates of Lactic Acid Bacteria From Fermented Fish Product, Budu

    Liasi, S. A.

    2009-01-01

    Full Text Available Three isolates of lactic acid bacteria (LAB from the fermented food product, Budu, were identified as genus lactobacillus (Lactobacillus casei LA17, Lactobacillus plantarum LA22 and L. paracasei LA02, and the highest population was Lb. paracasei LA02. The antibacterial agent produced by the isolates inhibited the growth of a range of gram-positive and gram-negative microorganisms. Antimicrobial sensitivity test to 18 different types of antibiotic were evaluated using the disc diffusion method. Inhibition zone diameter was measured and calculated from the means of five determinations and expressed in terms of resistance or susceptibility. All the LAB isolates were resistant to colestin sulphate, streptomycin, amikacin, norfloxacin, nalidixic acid, mecillinam, sulphanethoxazole/ trimethoprim, kanamycin, neomycin, bacitracin and gentamycin but susceptible to erythromycin, penicillin G, chloramphenicol, tetracycline, ampicillin and nitrofurantion.

  6. In vitro antifungal potentials of bioactive compound oleic acid, 3-(octadecyloxy) propyl ester isolated from Lepidagathis cristata Willd. (Acanthaceae) inflorescence

    Maghdu Nainamohamed Abubacker; Palaniyappan Kamala Devi

    2014-01-01

    Objective: To identify bioactive compound oleic acid, 3-(octadecyloxy) propyl ester from Lepidagathis cristata Willd. (L. cristata) and to assess antifungal potentials of the isolated compound. Methods: Aqueous extracts of L. cristata inflorescence were used for this study. The major bioactive compound isolated was tested for antifungal activities. Results: The major bioactive compound oleic acid, 3-(octadecyloxy) propyl ester was isolated from the inflorescence of L. cristata. The bioactive compound was tested for antifungal potentials and found to be highly effective to plant pathogenic fungi Colletotrichum fulcatum NCBT 146, Fusarium oxysporum NCBT 156 and Rhizoctonia solani NCBT 196 as well as for the human pathogenic fungi Curvularia lunata MTCC 2030 and Microsporum canis MTCC 2820. Conclusions: The results justify the antifungal potentials of both plant and human pathogenic fungi. The plant bioactive compound will be helpful in herbal antifungal formulations.

  7. Diversity of lactic acid bacteria isolated from Brazilian water buffalo mozzarella cheese.

    Silva, Luana Faria; Casella, Tiago; Gomes, Elisangela Soares; Nogueira, Mara Correa Lelles; De Dea Lindner, Juliano; Penna, Ana Lúcia Barretto

    2015-02-01

    The water buffalo mozzarella cheese is a typical Italian cheese which has been introduced in the thriving Brazilian market in the last 10 y, with good acceptance by its consumers. Lactic acid bacteria (LAB) play an important role in the technological and sensory quality of mozzarella cheese. In this study, the aim was to evaluate the diversity of the autochthones viable LAB isolated from water buffalo mozzarella cheese under storage. Samples were collected in 3 independent trials in a dairy industry located in the southeast region of Brazil, on the 28th day of storage, at 4 ºC. The LAB were characterized by Gram staining, catalase test, capacity to assimilate citrate, and production of CO2 from glucose. The diversity of LAB was evaluated by RAPD-PCR (randomly amplified polymorphic DNA-polymerase chain reaction), 16S rRNA gene sequencing, and by Vitek 2 system. Twenty LAB strains were isolated and clustered into 12 different clusters, and identified as Streptococcus thermophilus, Enterococcus faecium, Enterococcus durans, Leuconostoc mesenteroides subsp. mesenteroides, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus helveticus. Enterococcus species were dominant and citrate-positive. Only the strains of L. mesenteroides subsp. mesenteroides and L. fermentum produced CO2 from glucose and were citrate-positive, while L. casei was only citrate positive. This is the first report which elucidates the LAB diversity involved in Brazilian water buffalo mozzarella cheese. Furthermore, the results show that despite the absence of natural whey cultures as starters in production, the LAB species identified are the ones typically found in mozzarella cheese. PMID:25597646

  8. Isolation and Molecular Screening of Glucansucrase Gene Harboring-Lactic Acid Bacteria

    Ajitya Kurnia Hermawati

    2010-04-01

    Full Text Available Exopolysaccharides (EPS have been possessed to be used in pharmaceutical, cosmetic and food industries. Lactic acid bacteria (LAB produce a wide variety of exopolysaccharides and have been well reported carrying sucrase genes glucansucrase/ glucosyltransferase (gtf and fructansucrase/fructosyltransferases (ftf, enzymes that are able to produce EPS. In this study, the isolation and screening of EPS producing-LAB (EPS-LAB were carried out on modified de Mann-Rogosa-Sharpe (MRS agar medium supplemented with 10% of sucrose on LAB isolated from various unique sugar containing-foods and -beverages originated from local sources. Besides obtaining EPS-LAB, this study aimed to screen for gtf gene as well as to molecular identify strains by using PCR technique. Degenerate primer pairs DegFor and DegRev which targeted the conserved region of gtf genes catalytic domain were used, whereas LABfw and LABrv were used to molecular identify strains using 16S rRNA gene. An approximately 660 base pairs (bp amplicons which targeted gtf gene were obtained from 13 out of 16 srains chosen. Moreover, from PCR of 16S rRNA gene identification on gtf positive strains result, all strains were molecular identified as LAB after DNA sequencing analysis of 700 bp amplicons by using blastn. A rare EPS-producing LAB were obtain from both foods and beverages i.e. Weissella. Results revealed that strains obtained in this study are potential sources for exploring novel sucrase gene/s and obtain unique EPS polymer product/s.

  9. Antibacterial and Antioxidant Activities of Acid and Bile Resistant Strains of Lactobacillus fermentum Isolated from Miang

    Srikanjana Klayraung

    2009-12-01

    Full Text Available Miang is a kind of traditional fermented tea leaves, widely consumed in northern Thailand as a snack. It contains several kinds of Lactobacilli spp. The aim of this study was to isolate strains of Lactobacillus fermentum from miang and to investigate their antibacterial and antioxidant activities. The agar spot and well assays were used for determination of antibacterial power. The antibacterial mechanism was investigated by cell morphologic change under scanning electron microscope (SEM. Antioxidant activity was studied by means of free radical scavenging and ferric reducing power assays. The acid and bile screening tests indicated that L. fermentum FTL2311 and L. fermentum FTL10BR presented antibacterial activity against several pathogenic bacteria: Listeria monocytogenes DMST 17303, Salmonella Typhi DMST 5784, Shigella sonnei DMST 561 (ATCC 11060and Staphylococcus aureus subsp. aureus DMST 6512 (ATCC 6538Ptm. The results from SEM suggested that the antibacterial action was due to the destruction of cell membrane which consequently caused the pathogenic cell shrinking or cracking. The antioxidant study suggested that both L. fermentum FTL2311 and L. fermentum FTL10BR strains could liberate certain substances that possessed antioxidant activity expressed as trolox equivalent antioxidant capacity (TEAC and equivalent concentration (EC values for free radical scavenging and reducing mechanisms, respectively. The supernatant of L. fermentum FTL2311 broth revealed TEAC and EC values of 22.54±0.12 and 20.63±0.17 µM.mg-1 respectively, whereas that of L. fermentum FTL10BR yielded TEAC and EC values of 24.09±0.12 and 21.26±0.17 µM.mg-1 respectively. These two strains isolated from miang present high potential as promising health-promoting probiotics.

  10. Identification, stress tolerance, and antioxidant activity of lactic acid bacteria isolated from tropically grown fruits and leaves.

    Fessard, Amandine; Bourdon, Emmanuel; Payet, Bertrand; Remize, Fabienne

    2016-07-01

    From 6 samples of tropically grown fruits and leaves, 10 lactic acid bacteria belonging Leuconostoc, Weissella, and Lactobacillus species were isolated and identified by 16S rRNA gene sequencing and (GTG)5 fingerprinting. Acidification kinetics determined from BHI broth cultures showed genus-related patterns. In particular, Weissella cibaria appeared to act as a potent acidifier. Tolerance of isolates to acid, oxidative, or salt stress was highly variable and strain dependent. Isolate S14 (Leuconostoc pseudomesenteroides) growth was not affected by the presence of 0.05% H2O2, while Lactobacillus spp. isolates (S17 and S29) were the most tolerant to pH 4.5. The growth of 4 isolates, S5 (Leuconostoc mesenteroides), S14 and S10 (Leuconostoc pseudomesenteroides), and S27 (W. cibaria), was not affected by 5% NaCl. Nutritional beneficial properties were examined through measurement of antioxidant activities of short-term fermented pineapple juice, such as LDL oxidation and polyphenol content, and through exopolysaccharide formation from sucrose. Two isolates, S14 and S27, increased the antioxidant capacity of pineapple juice. The robust capacity of W. cibaria and of Leuconostoc pseudomesenteroides for vegetable lactic fermentation aimed to ameliorate food nutritional and functional quality was highlighted. PMID:27197991

  11. Investigation of antibacterial activity of Lactic Acid Bacteria isolated from traditional kordish cheese in comparison with commercial strains

    Fereshteh Tofangsazan

    2013-12-01

    Full Text Available Background and Aim: The health benefits of lactic acid bacteria in human, especially their anti-pathogenic properties has been the focus of recent interests. The objective of this study was to investigate the antibacterial activity of lactic acid bacteria (LAB isolated from traditional Kurdish cheese against a few bacterial pathogens. Materials and Methods: The cell free culture supernatant of LAB isolated from Kurdish cheese which was treated with heat and NaOH were tested for their antibacterial activity by Agar Disk Diffusion method. Moreover, Minimum Inhibition Concentration and Co-aggregation of LAB against pathogens were determined. Each test was repeated for three times. Results: The LAB isolates, in comparison with commercial lactic acid bacteria, showed suitable antibacterial activity. Heating the bacterial supernatant eliminated its anti-bacterial property; however, alkali treatment did not have any effect. The Minimum Inhibition Concentration did not show significant differences between native and commercial lactic acid bacteria; however, the native LAB showed suitable co-aggregation with pathogens. Conclusion: Traditional lactic acid bacteria and their metabolites can inhibit growth of pathogens. This shows the positive role of LAB in human health which necessitates their increase usage as natural antimicrobial agent.

  12. Inositol monophosphate phosphatase genes of Mycobacterium tuberculosis

    Parish Tanya

    2010-02-01

    Full Text Available Abstract Background Mycobacteria use inositol in phosphatidylinositol, for anchoring lipoarabinomannan (LAM, lipomannan (LM and phosphatidylinosotol mannosides (PIMs in the cell envelope, and for the production of mycothiol, which maintains the redox balance of the cell. Inositol is synthesized by conversion of glucose-6-phosphate to inositol-1-phosphate, followed by dephosphorylation by inositol monophosphate phosphatases (IMPases to form myo-inositol. To gain insight into how Mycobacterium tuberculosis synthesises inositol we carried out genetic analysis of the four IMPase homologues that are present in the Mycobacterium tuberculosis genome. Results Mutants lacking either impA (Rv1604 or suhB (Rv2701c were isolated in the absence of exogenous inositol, and no differences in levels of PIMs, LM, LAM or mycothiol were observed. Mutagenesis of cysQ (Rv2131c was initially unsuccessful, but was possible when a porin-like gene of Mycobacterium smegmatis was expressed, and also by gene switching in the merodiploid strain. In contrast, we could only obtain mutations in impC (Rv3137 when a second functional copy was provided in trans, even when exogenous inositol was provided. Experiments to obtain a mutant in the presence of a second copy of impC containing an active-site mutation, in the presence of porin-like gene of M. smegmatis, or in the absence of inositol 1-phosphate synthase activity, were also unsuccessful. We showed that all four genes are expressed, although at different levels, and levels of inositol phosphatase activity did not fall significantly in any of the mutants obtained. Conclusions We have shown that neither impA, suhB nor cysQ is solely responsible for inositol synthesis. In contrast, we show that impC is essential for mycobacterial growth under the conditions we used, and suggest it may be required in the early stages of mycothiol synthesis.

  13. Alkaline phosphatase for immunocytochemical labelling: problems with endogenous enzyme activity.

    Bulman, A. S.; Heyderman, E

    1981-01-01

    Alkaline phosphatase may be used as a label for immunocytochemistry and can be demonstrated in tissue sections using the single step naphthol phosphate method. Endogenous enzyme activity may not be destroyed by fixation in formalin, formol alcohol, Carnoy's or Baker's solutions and should be inhibited before results are assessed. Either Bouin's solution or periodic acid followed by potassium borohydride are satisfactory inhibitor and do not adversely affect immunocytochemical results.

  14. Bioengineered protein phosphatase 2A: Update on need

    Rubiolo, Juan A.; López-Alonso, Henar; Alfonso, Amparo; Vega, Félix V.; Vieytes, Mercedes Rodríguez; Botana, Luis M

    2013-01-01

    Harmful algal blooms caused by phytoplankton can occur in all aquatic environments. Some of the algae present in these blooms are capable of producing extremely potent toxins. Due to climate change and eutrophication, harmful algal blooms are increasing on a global scale. One kind of toxin producing algae are those that produce okadaic acid, its derivatives (dinophysistoxin-1 and 2), and microcystins. These toxins are potent inhibitors of protein phosphatase 2A, so this protein is used to det...

  15. Methods to distinguish various types of protein phosphatase activity

    To distinguish the action of protein Tyr(P) and protein Ser(P)/Thr(P) phosphatases on 32P-labeled phosphoproteins in subcellular fractions different inhibitors and activators are utilized. Comparison of the effects of added compounds provides a convenient, indirect method to characterize dephosphorylation reactions. Protein Tyr(P) phosphatases are specifically inhibited by micromolar Zn2+ or vanadate, and show maximal activity in the presence of EDTA. The other class of cellular phosphatases, specific for protein Ser(P) and Thr(P) residues, are inhibited by fluoride and EDTA. In this class of enzymes two major functional types can be distinguished: those sensitive to inhibition by the heat-stable protein inhibitor-2 and not stimulated by polycations, and those not sensitive to inhibition and stimulated by polycations. Preparation of 32P-labeled Tyr(P) and Ser(P) phosphoproteins also is presented for the direct measurement of phosphatase activities in preparations by the release of acid-soluble [32P]phosphate

  16. Anti-hepatoma activity and mechanism of ursolic acid and its derivatives isolated from Aralia decaisneana

    Ze Tian; Geng Lin; Rui-Xia Zheng; Feng Huang; Meng-Su Yang; Pei-Gen Xiao

    2006-01-01

    AIM: To investigate the anti-tumor activity of ursolic acid (UA) and its derivatives isolated from Aralia decaisneana on hepatocellular carcinoma both in vitro andin vivo.METHODS: In vivo cytotoxicity was first screened by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. Morphological observation, DNA ladder, flow cytometry analysis, Western blot and real time PCR were employed to elucidate the cytotoxic mechanism of UA. Implanted mouse hepatoma H22 was used to evaluate the growth inhibitory effect of UA in vivo.RESULTS: UA could significantly inhibit the proliferation of HepG2 and its drug-resistance strain, R-HepG2 cells,but had no inhibitory effect on primarily cultured normal mouse hepatocytes whereas all the six derivatives of UA could not inhibit the growth of all tested cell lines. Further study on mechanism demonstrated that apoptosis and G0/G1 arrest were involved in the cytotoxicity and cleavage of poly-(ADP-ribose)-polymerase (PARP). Downregulation of cyclooxygenase-2(COX-2) protein and upregulation of heat shock protein (HSP) 105 mRNA correlated to the apoptosis of HepG2cells treated with UA. In addition, UA also could inhibit the growth of H22 hepatoma in vivo.CONCLUSION: UA is a promising anti-tumor agent, but further work needs to be done to improve its solubility.

  17. Isolation and partial characterization of halotolerant lactic acid bacteria from two Mexican cheeses.

    Morales, Fredy; Morales, Jesús I; Hernández, César H; Hernández-Sánchez, Humberto

    2011-07-01

    Isolated strains of halotolerant or halophilic lactic acid bacteria (HALAB) from Cotija and doble crema cheeses were identified and partially characterized by phenotypic and genotypic methods, and their technological abilities were studied in order to test their potential use as dairy starter components. Humidity, a(w), pH, and salt concentration of cheeses were determined. Genotypic diversity was evaluated by randomly amplified polymorphic DNA-polymerase chain reaction. Molecular identification and phylogenetic reconstructions based on 16S rRNA gene sequences were performed. Additional technological abilities such as salt tolerance, acidifying, and proteolytic and lipolytic activities were also investigated. The differences among strains reflected the biodiversity of HALAB in both types of cheeses. Lactobacillus acidipiscis, Tetragenococcus halophilus, Weissella thailandensis, and Lactobacillus pentosus from Cotija cheese, and L. acidipiscis, Enterococcus faecium, Lactobacillus plantarum, Lactobacillus farciminis, and Lactobacillus rhamnosus from doble crema cheese were identified based on 16S rRNA. Quantitative and qualitative assessments showed strains of T. halophilus and L. plantarum to be proteolytic, along with E. faecium, L. farciminis, and L. pentosus to a lesser extent. Lipolytic activity could be demonstrated in strains of E. faecium, L. pentosus, L. plantarum, and T. halophilus. Strains belonging to the species L. pentosus, L. plantarum, and E. faecium were able to acidify the milk media. This study evidences the presence of HALAB that may play a role in the ripening of cheeses. PMID:21327742

  18. The metabolism of arachidonic acid in isolated perfused fetal and neonatal rabbit lungs

    The developmental pattern of fetal and neonatal rabbit lungs to metabolize arachidonic acid (AA) to different cyclo-oxygenase products was studied in isolated rabbit lungs, which were perfused with Krebs bicarbonate buffer. 14C-AA (66 nmol) was injected into the pulmonary circulation and the nonrecirculating perfusion effluent was collected for four minutes. About ten per cent of the injected radioactivity was found in the 0-4 min perfusion effluent. The metabolites of AA in the effluent were analyzed by thin layer chromatography. The major metabolites of AA were PGE2 and its 15-keto-derivates, but also PGF2 alpha and its 15-keto-derivates, TXB2 and 6-keto-PGF1 alpha were found in the effluent. The most drastic developmental change was the increase in the amount of 15-keto-metabolites of PGE2 from late fetal period to the lungs of one day old rabbits (1.8 fold increase between birth and first postnatal day). Smaller changes were detected in the amounts of other cyclo-oxygenase products

  19. Proteinase-producing halophilic lactic acid bacteria isolated from fish sauce fermentation and their ability to produce volatile compounds.

    Udomsil, Natteewan; Rodtong, Sureelak; Tanasupawat, Somboon; Yongsawatdigul, Jirawat

    2010-07-15

    Halophilic lactic acid bacteria were isolated from fish sauce mashes fermented at 1 to 12 months. Seven out of sixty-four isolates were selected according to their proteolytic activity and growth at 25% NaCl for characterization and investigation of volatile compound production. All selected isolates were Gram-positive cocci with pairs/tetrads and grew at 0-25% NaCl, pH 4.5-9.0. Results of 16S rRNA gene sequence analysis showed 99% homology to Tetragenococcus halophilus ATCC 33315. The restriction fragment length polymorphism (RFLP) patterns of all isolates were also similar to those of T. halophilus ATCC 33315. These isolates were, thus, identified as T. halophilus. All isolates hydrolyzed fish protein in the medium containing 25% NaCl. Intracellular aminopeptidase of 7 isolates exhibited the highest activity of 2.85-3.67 U/ml toward Ala-p-nitroanilide (Ala-pNA). T.halophilus strains MS33 and M11 showed the highest alanyl aminopeptidase activity (Phalophilus MS33 and MRC5-5-2 were 1-propanol, 2-methylpropanal, and benzaldehyde, corresponding to major volatile compounds in fish sauce. T.halophilus appeared to play an important role in volatile compound formation during fish sauce fermentation. PMID:20541276

  20. Solid and liquid media for isolating and cultivating acidophilic and acid-tolerant sulfate-reducing bacteria.

    Ňancucheo, Ivan; Rowe, Owen F; Hedrich, Sabrina; Johnson, D Barrie

    2016-05-01

    Growth media have been developed to facilitate the enrichment and isolation of acidophilic and acid-tolerant sulfate-reducing bacteria (aSRB) from environmental and industrial samples, and to allow their cultivation in vitro The main features of the 'standard' solid and liquid devised media are as follows: (i) use of glycerol rather than an aliphatic acid as electron donor; (ii) inclusion of stoichiometric concentrations of zinc ions to both buffer pH and to convert potentially harmful hydrogen sulphide produced by the aSRB to insoluble zinc sulphide; (iii) inclusion of Acidocella aromatica (an heterotrophic acidophile that does not metabolize glycerol or yeast extract) in the gel underlayer of double layered (overlay) solid media, to remove acetic acid produced by aSRB that incompletely oxidize glycerol and also aliphatic acids (mostly pyruvic) released by acid hydrolysis of the gelling agent used (agarose). Colonies of aSRB are readily distinguished from those of other anaerobes due to their deposition and accumulation of metal sulphide precipitates. Data presented illustrate the effectiveness of the overlay solid media described for isolating aSRB from acidic anaerobic sediments and low pH sulfidogenic bioreactors. PMID:27036143