Biotechnology and Bioprocess Engineering:BBE

  • 제7권6호
  • /
  • Pages.375-379
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  • 2002
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Purification and Characterization of Methyl Mercaptan Oxidase from Thiobacillus thioparus for Mercaptan Detection

  • Lee, Hyun-Ho (LG Institute of Environment, Safety & Health, Yonsei Engineering Research Center) ;
  • Kim, Sang-Joon (Department of Chemical and Biomolecular Engineering, KAIST) ;
  • Shin, Hyun-Jae (EnzBank, Inc.) ;
  • Park, Ji-Yeon (Department of Chemical and Biomolecular Engineering, KAIST) ;
  • Yang, Ji-Won (Department of Chemical and Biomolecular Engineering, KAIST)
  • 발행 : 2002.12.01
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초록

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$_4$Cl and (NH$_4$)$_2$SO$_4$, but was unaffected by either KCl or NaCl at less than 200 mM. With K$_2$SO$_4$, 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.

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