Purification and characterization of methyl mercaptan oxidase from Thiobacillus thioparus for mercaptan detection

被引：11

作者：

Lee H.-H. ^{[1
]}

Kim S.-J. ^{[2
]}

Shin H.-J. ^{[3
]}

Park J.-Y. ^{[2
]}

Yang J.-W. ^{[2
]}

机构：

[1] LG Institute of Environment, Safety and Health, Yonsei Engineering Research Center

[2] Department of Chemical and Biomolecular Engineering, KAIST

[3] EnzBank, Inc., 309 BVC, KRIBB

来源：

Biotechnology and Bioprocess Engineering | 2002年 / 7卷 / 6期

关键词：

Enzyme purification; Methyl mercaptan oxidase; Purpald; Thiobacillus thioparus;

D O I：

10.1007/BF02933525

中图分类号：

学科分类号：

摘要：

Methyl mercaptan oxidase was successfully induced in Thiobacillus thioparus TK-m using methyl mercaptan gas, and was purified for the detection of mercaptans. The purification procedure involved a DEAE (diethylaminoethyl)- Sephacel, or Superose 12, column chromatography with recovery yields of 47.5 and 48.5%, and specific activities of 374 and 1240.8 units/mg-protein, respectively. The molecular weight of the purified methyl mercaptan oxidase was 66.1kDa, as determined by SDS-PAGE. The extract, from gel filtration chromatography, oxidizes methyl mercaptan, producing formaldehyde, which can be easily detected by the purpald-coloring method. The optimized temperature for activity was found to be at 55°C. This enzyme was inhibited by both NH 4Cl and (NH4)2S04, but was unaffected by either KCl or NaCl at less than 200 mM. With K2S0 4, the activity decreased at 20 mM, but recovered at 150 mM. In the presence of methanol, full activity was maintained, but decreased in the presence of glycerin, ethanol and acetone 43, 78 and 75%, respectively.

引用

页码：375 / 379

页数：4

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