| 摘要 |
PURPOSE:To prepare creatinase useful for the determination of serum and creatinine, by culturing bacteria belonging to Bacillus genus and capable of producing creatinase. CONSTITUTION:Bacteria belonging to Bacillus genus and capable of producing creatinase [e.g. Bacillus sp. B-0618 (FERM-P No.4049)] are inoculated in a medium containing 0.5-1% of creatine, and cultured at 26-33 deg.C for 15-25hrs. The wet bacterial cells obtained by the solid/liquid separation of the cultured products, are suspended in a phosphoric acid buffer solution, etc., and subjected to a proper combination of conventional cell-treating procedures, e.g. lysozyme treatment, ultrasonic treatment, etc. The crude creatinase solution thus obtained is purified by conventional separation and purification procedures for proteins, enzymes, etc., and the purified product is freeze-dried in vacuum to obtain purified creatinase powder. |
