Purification and characterization of nitroreductase from red alkaliphilic bacterium Aquiflexum sp. DL6

Misal, Santosh A.; Bajoria, V. D.; Lingojwar, Devendra; Gawai, Kachru R.

doi:10.1134/s0003683813030125

Cited by 8 publications

(12 citation statements)

References 24 publications

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“…Similarly, a yield of 20% with 80 fold purification was recorded for nitrore ductase of Aquiflexum sp. DL6 [12]. A low specific activity of 1.47 U/mg with 10.1 fold increase was observed for the purification of NAD(P)H dependent nitroreductase of Pseudomonas sp.…”

Section: Resultsmentioning

confidence: 90%

“…C1 [8], Pseudomonas pseudoalcali genes JS45 [5], Aquiflexum sp. DL6 [12] and Pseudomonas sp. HK 6 [10].…”

Section: Resultsmentioning

confidence: 99%

“…reduction to 6 aminopendimethalin, and oxidative dealkylation to 3,4 dimethyl 2,6 dinitroaniline [15]. There are few reports on the purification and characterization of nitroreductases from microorganisms [5,8,11,12] but no information is available for the purification and characterization of pendimethalin nitroreductase from bacteria and fungi. The aim of this study was the purification and characterization of pendimethalin nitroreductase from Bacillus circulans.…”

mentioning

confidence: 98%

“…Some reports for the purification and biochemical properties of the nitroreductases obtainable from bacterial sources are available [1,[4][5][6][7][8][9][10][11][12]. Ammonium sulfate is often used as a precipitating agent for nitroreductases, whereas ion exchange chro matography and/or gel filtration chromatography are used to purify various nitroreductases [5,8,11,12].…”

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confidence: 99%

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Purification and properties of pendimethalin nitroreductase from Bacillus circulans

More¹,

Tallur²,

Ninnekar

et al. 2015

Appl Biochem Microbiol

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A pendimethalin nitroreductase was purified from Bacillus circulans to 52 fold increase with a recovery of 21.1% and had a specific activity of 4.16 U/mg. The molecular weight was 27 kDa. The optimum pH and temperature were 7.5 and 35°C, respectively. Fe 2+ , Ca 2+ , Mg 2+ and Mn 2+ did not cause any effect on the enzyme activity, whereas Ag + , Hg 2+ significantly inhibited it. The pendimethalin nitroreductase from B. circulans required NADPH as cofactor. It was not affected by GSH, DTT, L Cys and β mercaptoethanol but inhibited by p hydroxymercuribenzoate, N ethylmaleimide, and iodoacetamide. EDTA, 1,10 phenan throline and α,α' dipyridyl had a moderate effect on the enzyme activity. The nitroreductase reduced all the tested nitroaromatic compounds but was more active towards pendimethalin and trifluralin. The K M and V max for the enzyme reaction using pendimethalin as a substrate were 4.35 μM and 620 μmoles mg -1 min -1 , respectively. The broad substrate specificity towards dinitroaniline herbicides suggested the enzyme to be used as a potent reducing agent of these toxic compounds. This is the first report for the purification and charac terization of pendimethalin nitroreductase from any bacterial or fungal species.

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Section: Resultsmentioning

confidence: 90%

“…C1 [8], Pseudomonas pseudoalcali genes JS45 [5], Aquiflexum sp. DL6 [12] and Pseudomonas sp. HK 6 [10].…”

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 98%

mentioning

confidence: 99%

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Purification and properties of pendimethalin nitroreductase from Bacillus circulans

More¹,

Tallur²,

Ninnekar

et al. 2015

Appl Biochem Microbiol

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“…The azoreductase catalyze the reduction of azo dyes in presence of flavin and/or nicotinamide adenine dinucleotide/ nicotinamide adenine dinucleotide phosphate (NADH /NADPH) as an electron equivalent [13,14]. In our previous reports, we demonstrated that some of the nitro aromatic compounds can also be reduced by neutrophilic azoreductase by two electron reaction mechanism to its respective amines [15,16].…”

Section: Introductionmentioning

confidence: 99%

Biotransformation of Nitro Aromatic Compounds by Flavin-Free NADHAzoreductase

Misal¹

2015

J Bioremed Biodeg

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Nitro aromatic compounds are the potential toxic organic pollutant released into the environment and are often resistant to degradation under normal environmental conditions. The biotransformation and/or detoxification of these compounds can be possible by microbial azoreductase enzyme. Azoreductase enzyme has an ability to reduce the toxic nitro group to corresponding amino group. In present report, the flavin-free NADH azoreductase was isolated and purified from alkaliphilic bacteria Bacillus badius. The enzyme was purified by a combination of ammonium sulphate precipitation and size exclusion chromatography. The purified azoreductase has efficiently demonstrated both azoreductase and nitroreductase activities. The biochemical properties of the azoreductase including cofactor requirement, substrate specificity and enzyme inhibition have been studied. The biotransformation of some selected nitro aromatic compounds like 3-nitro benzoic acid, 4-nitro toluene, 3-nitro toluene and 1-chloro-2-nitro benzene by purified azoreductase was carried out at 37°C. The reduction products of these nitro aromatic compounds were analyzed by IR and NMR spectroscopy.

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Characterization and Purification of Membrane‐Bound Azoreductase From Azo Dye Degrading Shewanella sp. Strain IFN4

Imran

Negm

Hussain³

et al. 2016

CLEAN Soil Air Water

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Azo dyes can be degraded by azoreductases that achieve initial decolorization of the dye molecule through cleavage of the azo bond. Here we characterized membrane-bound azoreductase enzyme expressed by Shewanella sp. strain IFN4, which is a particularly efficient azo dye degrading bacterium. Initial studies showed that this bacterium was capable of decolorizing a wide variety of azo dyes including Reactive Black 5, Acid Red 88, Direct Red 81, Acid Yellow 19, and Disperse Orange 3. These structurally different azo dyes were also found to be reduced by the membrane-bound azoreductase showing the wider substrate specificity of the enzyme. Maximum enzyme activity was observed at pH 8 and 45°C with Reactive Black 5 as a substrate, and was stimulated on the addition of flavin or quinone compounds. The azoreductase was partially purified using a combination of ammonium sulfate precipitation followed by anion exchange chromatography. The cell membrane azoreductase was identified by non-denaturing gel electrophoresis and was shown to have a molar mass of 33 AE 0.5 kDa. Analysis of protein fragments by mass spectrometry showed high similarity with a Na(þ)translocating NADH-quinone reductase, previously identified in different species of the genus Shewanella. This enzyme is the first to be identified as a primary functional membrane-bound azoreductase used by members of the genus Shewanella to degrade azo dyes.Abbreviations: AQDS, anthraquinone-2,6-disulfonic acid; AQS, anthraquinone-2-sulfonic acid; AR 88, acid red 88; AY 19, acid yellow 19; CID, collision-induced dissociation; DO 3, disperse orange 3; DR 81, direct red 81; DTT, dithiothreitol; EDTA, ethylenediaminetetraacetic acid; MudPIT, multidimensional protein identification technology; Nano-LC/MS, nano liquid chromatography-mass spectrometery; NADH, nicotinamide adenine dinucleotide (reduced); RB 4, reactive blue 4; RB 5, reactive black 5; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis. 1523

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Purification and characterization of nitroreductase from red alkaliphilic bacterium Aquiflexum sp. DL6

Cited by 8 publications

References 24 publications

Purification and properties of pendimethalin nitroreductase from Bacillus circulans

Purification and properties of pendimethalin nitroreductase from Bacillus circulans

Biotransformation of Nitro Aromatic Compounds by Flavin-Free NADHAzoreductase

Characterization and Purification of Membrane‐Bound Azoreductase From Azo Dye Degrading Shewanella sp. Strain IFN4

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