Purification and Properties of Uricase from Enterobacter cloacae |
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Authors: | Yozo Machida Tora Nakanishi |
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Institution: | Tokyo Research Laboratory, Kyowa Hakko Kogyo Co., Ltd., Machida-shi, Tokyo 194, Japan |
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Abstract: | Uricase activity was found in Enterobacter cloacae KY3074 grown on guanine, hypoxanthine, uric acid, and xanthine media. The enzyme was purified from cells grown on uric acid as a source of nitrogen. The purification procedure included ammonium sulfate fractionation, gel filtration on Sephadex G-150, and column chromatography on DEAE-cellulose and DEAE-Sephadex. The enzyme had a molecular weight of about 105,000 and was specific for uric acid. The optimum pH was around 9.5, and the activity was inhibited by the presence of potassium cyanide, Ag+ or Cu2+. This uricase can be used for estimation of uric acid. |
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