Purification and Characterization of Chitinases from Bacillus licheniformis Using Auricularia auricular Colloidal Substance as a Substrate

2011 ◽  
Vol 233-235 ◽  
pp. 1014-1017
Author(s):  
Yong Jun Zhang ◽  
Qiao Xian ◽  
Jie Min He ◽  
Meng Yao Zhao
Keyword(s):  
Carbon Source ◽  
Bacillus Licheniformis ◽  
Sole Carbon Source ◽  
Culture Supernatant ◽  
Purification And Characterization ◽  
Medium Ph ◽  
Molecular Weights ◽  
Sds Page ◽  
Optimized Conditions

A chitinase was purified from the culture supernatant of Bacillus licheniformis with Auricularia auricular colloidal substance as the sole carbon source. The optimized conditions of this species strain for the production of chitinases were found to be when the culture was shaken at 37°C for 3 days in medium (pH 7) containing 0.05% KH2PO4,0.5%NaNO3, 0.05% MgSO4·7H2O and 1.5% Auricularia auricular colloidal substance powder(w/v). The molecular weights of the chitinases determined by SDS-PAGE were approximately 52kDa. The chitinases was inhibited highly by Mg2+ and PMSF, whereas Co2+ and Mn2+ could increase highly the activity of the chitinase.

Acid and alkaline phosphatases of Capnocytophaga species. II. Isolation, purification, and characterization of the enzymes from Capnocytophaga ochracea

1983 ◽  
Vol 29 (10) ◽  
pp. 1361-1368 ◽  
Author(s):  
Thomas P. Poirier ◽  
Stanley C. Holt
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Alkaline Phosphatases ◽  
Purification And Characterization ◽  
Molecular Weights ◽  
Sds Page ◽  
Kinetics Of

Capnocytophaga ochracea acid (AcP; EC 3.1.3.2) and alkaline (AlP; EC 3.1.3.1) phosphatase was isolated by Ribi cell disruption and purified by sodium dodecyl sulphate – polyacrylamide gel electrophoresis (SDS–PAGE.) Both phosphatases eluted from Sephadex G-150 consistent with molecular weights (migration) of 140 000 and 110 000. SDS–PAGE demonstrated a 72 000 and 55 000 subunit molecular migration for AcP and AlP, respectively. The kinetics of activity of purified AcP and AIP on p-nitrophenol phosphate and phosphoseryl residues of the phosphoproteins are presented.

scholarly journals Biodegradation of malathion by Bacillus licheniformis strain ML-1

2016 ◽  
Vol 68 (1) ◽  
pp. 51-59 ◽  
Author(s):  
Sara Khan ◽  
Habiba Zaffar ◽  
Usman Irshad ◽  
Raza Ahmad ◽  
Abdul Khan ◽  
...  
Keyword(s):  
Carbon Source ◽  
Bacillus Licheniformis ◽  
Sole Carbon Source ◽  
Sole Source ◽  
Minimal Salt Medium ◽  
Organophosphate Pesticide ◽  
Soil Enrichment ◽  
Enrichment Technique ◽  
Malathion Carboxylesterase

Malathion, a well-known organophosphate pesticide, has been used in agriculture over the last two decades for controlling pests of economically important crops. In the present study, a single bacterium, ML-1, was isolated by soil-enrichment technique and identified as Bacillus licheniformis on the basis of the 16S rRNA technique. The bacterium was grown in carbon-free minimal salt medium (MSM) and was found to be very efficient in utilizing malathion as the sole source of carbon. Biodegradation experiments were performed in MSM without carbon source to determine the malathion degradation by the selected strain, and the residues of malathion were determined quantitatively using HPLC techniques. Bacillus licheniformis showed very promising results and efficiently consumed malathion as the sole carbon source via malathion carboxylesterase (MCE), and about 78% malathion was degraded within 5 days. The carboxylesterase activity was determined by using crude extract while using malathion as substrate, and the residues were determined by HPLC. It has been found that the MCE hydrolyzed 87% malathion within 96 h of incubation. Characterization of crude MCE revealed that the enzyme is robust in nature in terms of organic solvents, as it was found to be stable in various concentrations of ethanol and acetonitrile. Similarly, and it can work in a wide pH and temperature range. The results of this study highlighted the potential of Bacillus licheniformis strain ML-1 as a biodegrader that can be used for the bioremediation of malathion-contaminated soil.

Induction, purification, and characterization of two extracellular α-L-arabinofuranosidases from Fusarium oxysporum

2003 ◽  
Vol 49 (10) ◽  
pp. 639-644 ◽  
Author(s):  
Gianni Panagiotou ◽  
Evagelos Topakas ◽  
Lina Economou ◽  
Dimitris Kekos ◽  
Basil J Macris ◽  
...  
Keyword(s):  
Carbon Source ◽  
Sugar Beet ◽  
Fusarium Oxysporum ◽  
Enzyme Induction ◽  
Enzyme Purification ◽  
Sole Carbon Source ◽  
Purification And Characterization ◽  
Optimal Activity ◽  
Molecular Masses

In the presence of L-arabinose as sole carbon source, Fusarium oxysporum produces two α-L-arabinofuranosidases (ABFs) named ABF1 and ABF2, with molecular masses of 200 and 180 kDa, respectively. The two F. oxysporum proteins have been purified to homogeneity. The purified enzymes are composed of three equal subunits and are neutral proteins with pIs of 6.0 and 7.3 for ABF1 and ABF2, respectively. With p-nitrophenyl α-L-arabinofuranoside (pNPA) as the substrate, ABF1 and ABF2 exhibited Km values of 0.39 and 0.28 mmol·L–1, respectively, and Vmax values of 1.6 and 4.6 µmol·min–1·(mg of protein)–1, respectively, and displayed optimal activity at pH 6.0 and 50–60 °C. ABFs released arabinose only from sugar beet arabinan and not from wheat soluble and insoluble arabinoxylans. The enzymes were not active on substrates containing ferulic acid ester linked to C-5 and C-2 linkages of pNPA showing that phenolic substituents of pNPA sterically hindered the action of ABFs.Key words: α-L-arabinofuranosidase, enzyme purification, enzyme induction.

Purification and characterization of cycloisomaltotetraose-forming glucanotransferases from Agreia sp. D1110 and Microbacterium trichothecenolyticum D2006

2021 ◽  
Vol 85 (3) ◽  
pp. 600-610
Author(s):  
Akihiro Fujita ◽  
Akira Kawashima ◽  
Yuuki Mitsukawa ◽  
Noriaki Kitagawa ◽  
Hikaru Watanabe ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Culture Supernatant ◽  
Coupling Reaction ◽  
The Other ◽  
Purification And Characterization ◽  
Other Hand ◽  
Sds Page ◽  
Ph Range ◽  
Hydrolysis Of

ABSTRACT Glucanotransferases that can synthesize cyclo-{→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→} (CI4) from dextran were purified to homogeneity from the culture supernatant of Agreia sp. D1110 and Microbacterium trichothecenolyticum D2006. The molecular mass of both enzymes was estimated to be 86 kDa by SDS-PAGE. The glucanotransferase, named CI4-forming enzyme, from Agreia sp. exhibited the highest activity at pH 6.0 and 40 °C. The enzyme was stable on the pH range of 4.6-9.9 and up to 40 °C. On the other hand, the enzyme from M. trichothecenolyticum exhibited the highest activity at pH 5.7 and 40 °C. The enzyme was stable on the pH range of 5.0-6.9 and up to 35 °C. Both enzymes catalyzed 4 reactions, namely, intramolecular α-1,6-transglycosylation (cyclization), intermolecular α-1,6-transglycosylation, hydrolysis of CI4, and coupling reaction. Furthermore, the CI4-forming enzyme produced CI4 from α-1,6-linked glucan synthesized from starch by 6-α-glucosyltransferase. These findings will enable the production of CI4 from starch.

Purification and characterization of an exochitinase from Bacillus thuringiensis subsp. aizawai and its action against phytopathogenic fungi

2006 ◽  
Vol 52 (7) ◽  
pp. 651-657 ◽  
Author(s):  
Luis Morales de la Vega ◽  
J Eleazar Barboza-Corona ◽  
Maria G Aguilar-Uscanga ◽  
Mario Ramírez-Lepe
Keyword(s):  
Bacillus Thuringiensis ◽  
Sodium Dodecyl ◽  
Sclerotium Rolfsii ◽  
Phytopathogenic Fungi ◽  
Purification And Characterization ◽  
Chitinolytic Enzyme ◽  
Bacillus Thuringiensis Subsp ◽  
Sds Page ◽  
Fusarium Sp

A chitinolytic enzyme from Bacillus thuringiensis subsp. aizawai has been purified and its molecular mass was estimated ca. 66 kDa by sodium dodecyl sulfate – polyacryamide gel electrophoresis (SDS–PAGE). The enzyme was able to hydrolyze chitin to chitobiosides but not carboxymethylcellulose, cellulose, pullulan, and laminarin. Optimal pH and temperature were detected at 6 and 50 °C, respectively. Stability, in the absence of substrate, was observed at temperatures less than 60 °C and pH between 5 and 8. Enzyme activity was significantly inhibited by K+ and EDTA and completely inhibited by Hg2+. Purified chitinase showed lytic activity against cell walls from six phytopathogenic fungi and inhibited the mycelial growth of both Fusarium sp. and Sclerotium rolfsii. The biocontrol efficacy of the enzyme was tested in the protection of bean seeds infested with six phytopathogenic fungi.Key words: chitinase, Bacillus thuringiensis, purification, phytopathogenic fungi.

Purification and characterization of tuberculin-active components from BCG

1975 ◽  
Vol 21 (12) ◽  
pp. 2019-2027
Author(s):  
M. Laguerre ◽  
R. Turcotte
Keyword(s):  
Physicochemical Properties ◽  
Guinea Pigs ◽  
Gel Filtration ◽  
Biologically Active ◽  
Polyacrylamide Gels ◽  
Active Components ◽  
Purification And Characterization ◽  
Molecular Weights ◽  
Antigenic Activity

The tuberculin activity of protoplasmic extracts isolated from living BCG was purified successively by gel filtration on Sephadex G-100 and G-75, and by electrophoresis on 7.5% and on gradient (6–18%) polyacrylamide gels. The tuberculin-active fractions, as determined in BCG-sensitized guinea pigs, were used as the starting material for each of the following fractionation steps.The physicochemical properties and the antigenic activity of the biologically active fractions have shown that a single component, or only a few ones with similar properties, possessed high tuberculin activity. These active components were proteins having relatively high molecular weights (about 72 000) and could behave as antigens.

scholarly journals Purification and Characterization of 2,6-β-d-Fructan 6-Levanbiohydrolase fromStreptomyces exfoliatus F3-2

2000 ◽  
Vol 66 (1) ◽  
pp. 252-256 ◽  
Author(s):  
Katsuichi Saito ◽  
Kazuya Kondo ◽  
Ichiro Kojima ◽  
Atsushi Yokota ◽  
Fusao Tomita
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Extracellular Enzyme ◽  
Molecular Weights ◽  
Sds Page ◽  
Monomeric Structure ◽  
Ph Range ◽  
Hydrolysis Of

ABSTRACT Streptomyces exfoliatus F3-2 produced an extracellular enzyme that converted levan, a β-2,6-linked fructan, into levanbiose. The enzyme was purified 50-fold from culture supernatant to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of this enzyme were 54,000 by SDS-PAGE and 60,000 by gel filtration, suggesting the monomeric structure of the enzyme. The isoelectric point of the enzyme was determined to be 4.7. The optimal pH and temperature of the enzyme for levan degradation were pH 5.5 and 60°C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50°C. The enzyme gave levanbiose as a major degradation product from levan in an exo-acting manner. It was also found that this enzyme catalyzed hydrolysis of such fructooligosaccharides as 1-kestose, nystose, and 1-fructosylnystose by liberating fructose. Thus, this enzyme appeared to hydrolyze not only β-2,6-linkage of levan, but also β-2,1-linkage of fructooligosaccharides. From these data, the enzyme from S. exfoliatus F3-2 was identified as a novel 2,6-β-d-fructan 6-levanbiohydrolase (EC 3.2.1.64 ).

scholarly journals Isolation and Characterization of a Novel p,p’-DDT-Degrading Bacterium: Aeromonas sp. Strain MY1 from Agricultural Soil

2020 ◽  
pp. 12-22
Author(s):  
Y. Murtala ◽  
B. C. Nwanguma ◽  
L. U. S. Ezeanyika
Keyword(s):  
Carbon Source ◽  
16S Rrna ◽  
Agricultural Soil ◽  
Sole Carbon Source ◽  
Phylogenetic Analyses ◽  
Inhibitory Effect ◽  
Aerobic Conditions ◽  
Degrading Bacterium ◽  
Isolation And Characterization

Background: Despite the banned on the use of dichlorodiphenyltrichloroethane (DDT) and other Persistent Organic Pollutants (POPs) by the Stockholm Convention for their toxicity, emerging shreds of evidence have indicated that DDT is, however, still in use in developing countries. This might increase the global burden of DDT contamination and its hazardous effects. Aim: This study focused on the isolation and characterization of p,p’-DDT-degrading bacterium from a tropical agricultural soil. Methodology: Standard isolation procedure was used for the screening and isolation of the strain. The 16S rRNA and phylogenetic analyses were used to identify the isolate and established protocols were followed to characterize the strain. Results: A new strain belonging to the genus Aeromonas was isolated from agricultural soil using minimal salt-p,p’-DDT enrichment medium. The 16S rRNA sequencing was used to identify the strain and the partial sequence was deposited in the NCBI GenBank as Aeromonas sp. Strain MY1. This mesophilic isolate was capable of utilizing up to 50 mgL-1 of p,p’-DDT as the sole carbon source at an optimum pH of 7.5 and optimum temperature of 35 °C within 120 h under aerobic conditions. Fe2+ (0.2 mgL-1) demonstrated a stimulatory effect on the p,p’-DDT degradation capacity by the strain MY1. However, Zn, Cu, Pb, Hg, Ag and Cr ions have demonstrated various patterns of inhibitory effect on the p,p’-DDT degradation capacity of the isolate at 0.2 mgL-1. The strain MY1 could be a promising candidate for the bioremediation of p,p’-DDT contaminant. Conclusion: Aeromonas sp. strain MY1 was capable of utilizing p,p’-DDT as a sole carbon source under aerobic conditions. The utilization capacity of the strain was influenced by some heavy metals. Fe was found to enhance the p,p’-DDT utilization capacity of the isolate at a lower concentration. While Zn, Cu, Pb, Hg, Ag and Cr showed various patterns of inhibitory effect.

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