Purification and Characterization of a Nonylphenol(NP)-degrading Enzyme from Bacillus cereus.Frankland

Abstract

Abstract: Abstract>An extracellular NP-degrading enzyme secreted by Bacillus cereus.Frankland was purified to homogeneity by a combination of ammonium sulfate precipitation, Phenyl-Sepharose hydrophobic-interaction chromatography and DEAE anion-exchange chromatography.On SDS(sodium dodecyl sulfate)-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a relative molecular mass of 58.3 kDa.The depolymerzation of subunits was accompanied with the loss of NP-degrading enzyme activity, and removing denaturing factors by dialysis could restore the dimer structure and enzymatic activity.The enzyme had an isoelectric point of 5.5 and an optimal temperature of 60℃, and was the most active at pH 6.0.The enzymatic activity was stable at pH 4-8 and inhibited by Cu²⁺.TenN-terminal amino acids were determined to be ASVNSIKIGY, demonstrating that the purified enzyme was a novel one.The hydrolysis pattern of the purified enzyme indicated that the NP-degrading enzyme was an endo NP-degrading enzyme.The extraordinary thermo-stability provided the enzyme with a good prospect to be used as a new tool for clean-production process for textile industry.

Key words: nonylphenol(NP), Bacillus cereus.Frankland, NP-degrading enzyme, purification, characterization

摘要： Abstract>An extracellular NP-degrading enzyme secreted by Bacillus cereus.Frankland was purified to homogeneity by a combination of ammonium sulfate precipitation, Phenyl-Sepharose hydrophobic-interaction chromatography and DEAE anion-exchange chromatography.On SDS(sodium dodecyl sulfate)-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a relative molecular mass of 58.3 kDa.The depolymerzation of subunits was accompanied with the loss of NP-degrading enzyme activity, and removing denaturing factors by dialysis could restore the dimer structure and enzymatic activity.The enzyme had an isoelectric point of 5.5 and an optimal temperature of 60℃, and was the most active at pH 6.0.The enzymatic activity was stable at pH 4-8 and inhibited by Cu²⁺.TenN-terminal amino acids were determined to be ASVNSIKIGY, demonstrating that the purified enzyme was a novel one.The hydrolysis pattern of the purified enzyme indicated that the NP-degrading enzyme was an endo NP-degrading enzyme.The extraordinary thermo-stability provided the enzyme with a good prospect to be used as a new tool for clean-production process for textile industry.

关键词: nonylphenol(NP), Bacillus cereus.Frankland, NP-degrading enzyme, purification, characterization

YANG Ge, ZHANG Ying, BAI Yanfen. Purification and Characterization of a Nonylphenol(NP)-degrading Enzyme from Bacillus cereus.Frankland [J]. , 2011, 19(4): 644-648.

杨革, 张营, 白艳芬. Purification and Characterization of a Nonylphenol(NP)-degrading Enzyme from Bacillus cereus.Frankland [J]. , 2011, 19(4): 644-648.

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