Screening of the Bacterium for Chlorobenzene Degradation and its Enzymatic Properties

2012 ◽  
Vol 610-613 ◽  
pp. 404-408
Author(s):  
Xian Niu ◽  
Cheng Ding ◽  
Jin Long Yan ◽  
Bai Ren Yang
Keyword(s):  
Enzyme Purification ◽  
Extracellular Enzymes ◽  
Ph Value ◽  
Morphological Characteristics ◽  
Enzymatic Properties ◽  
Purification Process ◽  
Optimum Temperature ◽  
16S Rdna Sequence ◽  
Sds Page ◽  
Optimum Ph

A dominant bacterium (LW13) for the degradation of chlorobenzene was selected from maturity sludge in a novel combined bio-filter polluted by chlorobenzene gas. Based on the morphological characteristics observation, physio-biochemical characteristics and 16S rDNA sequence homology analysis, strain LW13 was identified as Lysinibacillus fusiformis. Crude enzyme from the fermentation was extracted and their enzymatic properties were also investigated. Results showed that the degradation enzyme produced by the bacteria belong to extracellular enzymes. The purity of the enzyme was determined by SDS-PAGE gel electrophoresis and the molecular weight was found to be 52 kDa. The optimum pH value was about 8.0 with the optimum temperature of 45° C. Throughout the purification process, 85-fold of enzyme purification was achieved with the recovery of 20.69%.

scholarly journals Characterization of Biosynthetic Enzymes for Ectoine as a Compatible Solute in a Moderately Halophilic Eubacterium, Halomonas elongata

1999 ◽  
Vol 181 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Hisayo Ono ◽  
Kazuhisa Sawada ◽  
Nonpanga Khunajakr ◽  
Tao Tao ◽  
Mihoko Yamamoto ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Apparent Molecular Mass ◽  
Optimum Temperature ◽  
Sds Page ◽  
Halomonas Elongata ◽  
Optimum Ph

ABSTRACT 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. The biosynthetic pathway of ectoine from aspartic β-semialdehyde (ASA), in Halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-Diaminobutyrate (DABA) aminotransferase catalyzed reversively the first step of the pathway, conversion of ASA to DABA by transamination with l-glutamate. This enzyme required pyridoxal 5′-phosphate and potassium ions for its activity and stability. The gel filtration estimated an apparent molecular mass of 260 kDa, whereas molecular mass measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 44 kDa. This enzyme exhibited an optimum pH of 8.6 and an optimum temperature of 25°C and had Km s of 9.1 mM forl-glutamate and 4.5 mM for dl-ASA. DABA acetyltransferase catalyzed acetylation of DABA to γ-N-acetyl-α,γ-diaminobutyric acid (ADABA) with acetyl coenzyme A and exhibited an optimum pH of 8.2 and an optimum temperature of 20°C in the presence of 0.4 M NaCl. The molecular mass was 45 kDa by gel filtration. Ectoine synthase catalyzed circularization of ADABA to ectoine and exhibited an optimum pH of 8.5 to 9.0 and an optimum temperature of 15°C in the presence of 0.5 M NaCl. This enzyme had an apparent molecular mass of 19 kDa by SDS-PAGE and a Km of 8.4 mM in the presence of 0.77 M NaCl. DABA acetyltransferase and ectoine synthase were stabilized in the presence of NaCl (>2 M) and DABA (100 mM) at temperatures below 30°C.

scholarly journals Purification and characterization of beta-1, 3-glucanase from the secretion of Simira glaziovii colleters (Rubiaceae)

2006 ◽  
Vol 49 (6) ◽  
pp. 881-888 ◽  
Author(s):  
Felipe Almeida Vieira ◽  
Maura da Cunha ◽  
Denise Espellet Klein ◽  
André de Oliveira Carvalho ◽  
Valdirene Moreira Gomes
Keyword(s):  
Molecular Mass ◽  
Purification Process ◽  
Purification And Characterization ◽  
Chromatographic Methods ◽  
Sds Page ◽  
Optimum Ph

In this study, beta-1,3-glucanase was isolated from Simira glaziovii secretion. The purification process was achieved by a combination of chromatographic methods and was analyzed by SDS-PAGE. The purified enzyme presented an estimated molecular mass of 35 kDa. The optimum pH of enzyme was 5.2

Inhibition by Substrates of a Coniferyl Alcohol Dehydrogenase Purified from Sugarcane Stalks

2020 ◽  
Vol 15 (3) ◽  
pp. 206-214
Author(s):  
Borja Alarcón ◽  
Roberto de Armas ◽  
Carlos Vicente ◽  
María E. Legaz
Keyword(s):  
Alcohol Dehydrogenase ◽  
Ph Value ◽  
Coniferyl Alcohol ◽  
Kinetic Properties ◽  
Optimum Temperature ◽  
Alcohol Dehydrogenases ◽  
Sinapyl Alcohol ◽  
Optimum Ph ◽  
Km Value ◽  
Similar Affinity

Aims and Objectives: This study aimed to characterize a coniferyl alcohol dehydrogenase from sugarcane stalks. Also, the purification of CAD from sugarcane stalks was also carried out to study kinetic properties and substrate specificity. Background: Sugarcane plants contain an alcohol dehydrogenase able to reduce both coniferyl and sinapyl aldehydes to their correspondent alcohols, although there are reasonable grounds for suspecting that these are two distinct enzymes. Methods: The enzyme, coniferyl alcohol dehydrogenase was 125-fold purified from sugarcane stalks. Its activity was estimated by HPLC by calculating the amount of product formed. Results: The enzyme showed an optimum pH value of 7.9, at an optimum temperature of 20-22°C and a molecular mass of 48 kDa. The Km value for coniferyl alcohol was 3.03 µM and the enzyme was shown to be inhibited by an excess of the substrate from 17 µM. This dehydrogenase showed a similar affinity to sinapyl alcohol (Km 1.78 µM). Conclusions: This paper provides circumstantial evidence about the existence of two different alcohol dehydrogenases, specific to each of the substrates.

scholarly journals Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme

2014 ◽  
Vol 32 (No. 1) ◽  
pp. 48-53 ◽  
Author(s):  
K.C. Blanco ◽  
F.F. de Moraes ◽  
N.S. Bernardi ◽  
M.H.P.B. Vettori ◽  
R. Monti ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Maximum Velocity ◽  
Temperature Stability ◽  
Cassava Starch ◽  
Optimum Temperature ◽  
Cyclodextrin Glycosyltransferase ◽  
Sds Page ◽  
Optimum Ph ◽  
Menten Constant ◽  
Cyclodextrin Production

The properties of a previously unknown enzyme, denominated cyclodextrin glycosyltransferase, produced from Bacillus lehensis, were evaluated using affinity chromatography for protein purification. Enzyme characteristics (optimum pH and temperature; pH and temperature stability), the influence of substances on the enzyme activity, enzyme kinetics, and cyclodextrin production were analysed. Cyclodextrin glycosyltransferase was purified up to 320.74-fold by affinity chromatography using β-cyclodextrin as the binder and it exhibited 8.71% activity recovery. This enzyme is a monomer with a molecular weight of 81.27 kDa, as estimated by SDS-PAGE. Optimum temperature and pH for cyclodextrin glycosyltransferase were 55°C and 8.0, respectively. The Michaelis-Menten constant was 8.62 g/l during maximum velocity of 0.858 g/l.h.  

Preparation of Genipin by Hydrolysis of Geniposide with Co-Immobilized Enzyme

2011 ◽  
Vol 236-238 ◽  
pp. 1793-1798 ◽  
Author(s):  
Hua Zheng Liang ◽  
He Chen ◽  
Jian Feng Wang ◽  
Yu Lan He
Keyword(s):  
High Performance ◽  
Immobilized Enzyme ◽  
Low Cost ◽  
Enzymatic Properties ◽  
High Yield ◽  
Cross Linking ◽  
Optimum Temperature ◽  
Environmental Friendly ◽  
Optimum Ph ◽  
Hydrolysis Of

Co-immobilize enzyme by cross-linking and embedding, optimize conditions for immobilizing, determinate the enzymatic properties of co-immobilized enzyme and study the methods for preparation of genipin using co-immobilized enzyme to hydrolyze geniposide. Optimized immobilizing conditions include glutaraldehyde concentration being 0.15%, cross-linking temperature being 20°C, cross-linking time being 2 hours, the activity of co-immobilized β-glucosidase and cell reaches to 65.33U/mg and the enzyme activity recovery being 52.63%. Enzymatic properties of co-immobilized enzyme are following: optimum temperature is 55°C and optimum pH is 5.0. The transformation experiments are carried out with co-immobilized enzyme. The results show that half-life of co-immobilized enzyme reaches around 40 days, higher than the normal immobilized enzyme. The conversion rate of geniposide is above 95% after 8 hours. The genipin is isolated, purified and recrystallized to reach more than 98% of purity by High Performance Liquid Chromatography. Advantages to prepare genipin using co-immobilized enzyme include low cost, high yield, environmental friendly and easy to manufacturing.

scholarly journals Purification, Characterization and De-Staining Potentials of a Thermotolerant Protease Produced by Fusarium oxysporum

2019 ◽  
Vol 64 (4) ◽  
pp. 539-547
Author(s):  
Mohammed Inuwa Ja’afaru ◽  
Konjerimam Ishaku Chimbekujwo ◽  
Obinna Markraphael Ajunwa
Keyword(s):  
Enzyme Activity ◽  
Fusarium Oxysporum ◽  
Treatment Time ◽  
Specific Activity ◽  
Total Yield ◽  
Tween 20 ◽  
Optimum Temperature ◽  
Industrial Enzymes ◽  
Sds Page ◽  
Optimum Ph

Proteases are important industrial enzymes and fungi prove to be good sources of such enzymes. Purification techniques are however necessary for increased specificity in activity and better industrial value. Based on this, a protease produced by a Fusarium oxysporum was purified to homogeneity by Sephadex G-200 column and α–casein agarose chromatography. The enzyme had a molecular weight of 70 kDa in SDS-PAGE. Purified Fusarium oxysporum protease had a specific activity of 93.88 U/mg protein. The purification magnitude was 7.7 and the total yield was 20 %. Purified protease had an optimum pH of 5.0 while the optimum temperature was 40 °C. The enzyme was also thermotolerant (approximately 100 % at 40 °C for 2 h). The enzyme activity was stimulated by surfactants and metal ions like, Tween-20 and Mg2+. Enzyme activity was inhibited in presence of PMSF and EDTA. Casein was found to be the best substrate for protease activity of Fusarium oxysporum FWT1. Protease were tested upon blood stain for de-clotting of blood and was found to exhibit good de-clotting and de-staining activity after 15 minutes treatment time.

scholarly journals A Metalloaminopeptidase from Flavobacterium breve: Characterization and Molecular Cloning

2019 ◽  
Vol 1 (2) ◽  
pp. 11-17
Author(s):  
Sreekumar Othumpangat ◽  
Kiyoshi Hayashi
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Maximum Activity ◽  
Optimum Temperature ◽  
High Affinity ◽  
Aeromonas Caviae ◽  
Sds Page ◽  
Optimum Ph ◽  
Tail Pcr

Aminopeptidase from Flavobacterium breve, was purified by a three step FPLC column chromatography to homogeneity from the culture filtrate. The aminopeptidase gene was cloned by using TAIL-PCR technique. The gene encodes for a polypeptide composed of 497 amino acids with a theoretical molecular weight of 58 kDa. SDS-PAGE detection revealed that the protein is of 52 kDa. The native enzyme showed high affinity to Leu-pNA (km 0.0515 mM), and kcat /km of 88.8 s-1mM-1 . The enzyme had an optimum pH 7.5 and was stable from pH 6 to 9. The purified aminopeptidase was stable up to 60 oC and the optimum temperature for the maximum activity was at 70 oC. The amino acid sequence showed 47% identity to aminopeptidase of Aeromonas caviae (family M14), a Zn2+ dependent metallozyme.

Characterization of a Laccase from White Rot Fungus Cerrena unicolor

2013 ◽  
Vol 442 ◽  
pp. 120-124
Author(s):  
Bei Bei Zhang ◽  
Yong Jian Shen ◽  
Chang Xu ◽  
Ying Li ◽  
Ming Hu Jiang
Keyword(s):  
Dye Decolorization ◽  
White Rot ◽  
White Rot Fungus ◽  
Optimum Temperature ◽  
Complex Chemical ◽  
Chemical Structures ◽  
Cerrena Unicolor ◽  
Sds Page ◽  
Optimum Ph

Dyes are usually difficult to be decolorized due to their complex chemical structures. In this work, laccase was purified from the white rot fungus Cerrena unicolor to evaluate its application in dye decolorization. SDS-PAGE analysis showed the purified laccase to be a monomeric protein of 63.7 kDa. The optimum pH for the oxidation of 2,2-azinobis-(3-ethylbenzthiaoline-6-sufonic acid) (ABTS) was 2.2 and the optimum temperature was 50°C. The activity of the purified enzyme was strongly inhibited by sodium azide and partially inhibited by cysteine, dithiothreitol. The Km values of the purified laccase for the substrate ABTS, syringaldazine and 2,6-dimethoxyphenol were 0.217, 0.306 and 0.199 mmol/L.

scholarly journals Isolasi Bakteri dan Karakterisasi Protease dari Sumber Air Rawa Indralaya

2012 ◽  
Vol 14 (1) ◽  
pp. 114
Author(s):  
Ace Baehaki ◽  
Rinto Rinto ◽  
Agus Ramiadi
Keyword(s):  
Metal Ion ◽  
Optimum Temperature ◽  
Microbial Isolation ◽  
Isolation And Characterization ◽  
Sds Page ◽  
Optimum Ph

The aim of this study was to bacteria isolation and characterization proteases from water swamp. The water swamp samples collected fromIndralaya for microbial isolation. Three isolates showed proteolytic index >1.00. The optimum pH of extraceluller proteases from A6S3,A4S3 and A15S3 were 7.5; 8.0; 8.0, respectively. The optimum temperature of A6S3, A4S3 and A15S3 protease were 40; 50; 50 oC,respectively. Effect of metal ion (Fe, K, Mn, and Zn) shown Fe and K were inhibit protease A6S3, all metal ion were inhibit protease A4S3and K, Mn and Zn inhibit protease A15S3. Study on the effect of metals ion and spesific inhibitors indicated that all protease weremetaloprotease. Their moleculer weights were determined by using SDS-PAGE and zymogram technique for A6S3 isolate were 70, 88, 106and 121 kD respectively. Whereas for isolate A4S3 was 138 kD and isolate A15S3 was 131 kD.

Study on Enzymatic Characteristics of Chitin Deacetylase

2011 ◽  
Vol 236-238 ◽  
pp. 996-999 ◽  
Author(s):  
Jun Ang Liu ◽  
Yuan Hao He
Keyword(s):  
Inhibitory Effect ◽  
Enzymatic Properties ◽  
Optimum Temperature ◽  
Chitin Deacetylase ◽  
Activation Effect ◽  
High Productivity ◽  
Enzymatic Characteristics ◽  
Optimum Ph

Chitin deacetylase (CDA) catalyzes the conversion of chitin to chitosan by the deacetylation of N-acetyl-D-glucosamine residues. The results of the fundamental enzymatic properties of chitin deacetylase producing from the high productivity chitin deacetylase strain Z7 show that, the optimum temperature of strain Z7 was 40°C, the optimum pH was 6.5, under the concentration of 1mmol/L, Mg2+, K+, Ca2+and Fe2+had activation effect, of which the strongest activation effect was Mg2+. Pb2+, Cu2+and Mn2+had inhibitory effect, of which the strongest inhibitory effect was Mn2+.

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