Isolation and purification of methyl mercaptan oxidase fromRhodococcus rhodochrous for mercaptan detection |
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Authors: | Sang-Joon Kim Hyun-Jae Shin Yeu-Chun Kim Dae-Sil Lee Ji-Won Yang |
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Institution: | (1) Department of Chemical Engineering, KAIST, 305-701 Taejon, Korea;(2) EnzBank, Inc., KRIBB, 309 BVC, 305-333 Taejon, Korea |
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Abstract: | Methyl mercaptan oxidase was successfully induced fromRhodococcus rhodochrous IGTS8 using methyl mercaptan gas and purified to homogeneity for the detection of mecrcaptans. The purification procedure
involved DEAE-Sephacel and Superose 12 column chromatography with recovery yields of 85.8 and 83.3%, and a specific activity
of 92.7 and 303.4 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan, oxidase was determined
to be 64.5 kDa by SDS-PAGE. The extract from gel filtration chromatography oxidizes methyl mercaptan to produce formaldehyde,
which can be easily detected by the purpald-coloring method. Optimum temperature for activity was achieved at 60°C. This enzyme
was inhibited by both K2SO4 and NaCl at concentration of less than 100 mM and recovered to original activity at concentration of 200 mM. In the presence
of methanol, the activity decreased by 33%. |
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Keywords: | methyl mercaptan oxidase Rhodococcus rhodochrous purpald enzyme purification |
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