PURPOSE:To prepare sarcosine oxidase which can be used for the determination of creatinine in serum or urine, and is useful as an enzymatic clinical diagnostic agent, by the cultivation of bacterial belonging to Bacillus genus and having sarcosin oxidase-producing capability. CONSTITUTION:Sarcosine oxidase-producing bacteria belonging to Bacillus genus, e.g. Bacillus sp.B0618 (FERM-P No.4049), are inoculated in a nutrient medium, and cultured pref. in an aerated and agitated deep-tank fermentor. The cultivation product is separated into solid and liquid, and sarcosine oxidase is separated from the cells. Addition of creatine to the medium improves the yield of sarcosine oxidase. The culturing is conducted at 26-33 deg.C, for usually 15-25 hours. Sarcosine oxidase decomposes 1 mole of sarcosine into 1 mole of formaldehyde 1 mole of hydrogen peroxide, consuming 1 mole of water and 1 mole of oxygen.
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