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KMID : 0903519920350040332
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1992 Volume.35 No. 4 p.332 ~ p.333
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Production of cholesterol oxidase by Bacillus sphaericus
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Choi, Seung Il
Choi, Seung Chul/Son, Heung Soo
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Abstract
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It is well-known that the oxidation of cholesterol to cholest-4-en-3-one by cholesterol oxidase (cholesterol oxygen oxidoreductase EC 1.1.3.6) is the first step in the microbial decomposition of cholesterol. The application of cholesterol oxidase from microorganisms for the clinical determination of cholesterol in serum has become increasingly important. A bacterial strain capable of producing high level of intracellular cholesterol oxidase is isolated from several soil samples. The characteristics of the isolated strain No. 796 are identified to be Bacillus shaericus. The enzyme activity is increased when the microogranism is grown on cholesterol 0.3%. The maximal cholesterol oxidase production is observed in the medium containing 0.3% cholesterol, 0.25% glucose, 0.25% yeast extract, 0.25% NaCl, 0.025% K₂HPO₄, 0.025% KH₂PO₄ and 0.05% MgSO₄ with initial pH of 7.0. The optimal culture conditions for production of cholesterol oxidase are at 30℃ for 60 hrs at an agitation speed 150 rpm. The enzyme activity appears in the culture medium at the logarithmic phase, increases in parallel with cell growth and reaches the maximum at the early stationary phase.
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