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

›› 2011, Vol. 19 ›› Issue (4): 644-648.

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

杨革^1,2, 张营¹, 白艳芬²

1. College of Textiles Tianjin Polytechnic University, Tianjin 300160, China;
2. Key Lab of Biogeology and Environmental Geology of Ministry of Education, China University of Geosciences, Wuhan 430074, China

收稿日期:2010-04-07 修回日期:2011-04-18 出版日期:2011-08-28 发布日期:2011-08-28
通讯作者: YANG Ge,E-mail:yangge100@126.com
基金资助:
Supported by the Fund of Open Subject of Key Lab of Biogeology and Environmental Geology of Ministry of Education(BGEG1006)

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

YANG Ge^1,2, ZHANG Ying¹, BAI Yanfen²

1. College of Textiles Tianjin Polytechnic University, Tianjin 300160, China;
2. Key Lab of Biogeology and Environmental Geology of Ministry of Education, China University of Geosciences, Wuhan 430074, China

Received:2010-04-07 Revised:2011-04-18 Online:2011-08-28 Published:2011-08-28
Supported by:
Supported by the Fund of Open Subject of Key Lab of Biogeology and Environmental Geology of Ministry of Education(BGEG1006)

摘要/Abstract

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

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

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.

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.

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

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

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