疣孢漆斑菌(Myrothecium Verrucaria)J-1胆红素氧化酶的纯化及性质初步研究

Purification and Characteristics of Bilirubin Oxidase From Myrothecium Verrucaria J-1

从中国土样中筛选到一株能产生胆红素氧化酶的微生物(Myrothecium Verrucaria)J-1,培养后,分离纯化,最后经QAE—Sephadex A50柱层析,得到胆红素氧化酶比活为207.65 U/A 280nm,总产率为22.3％。纯酶紫外吸收峰为278 nm,凝胶电泳为单一色带。分子量估计为52000。它能迅速、特异地氧化胆红素为胆绿素,并进一步氧化成目前还不清楚的紫色化合物。最佳作用pH为7.0,最佳作用温度为40℃。

A strain (Myrothecium Verrucaria) J-1 producing bilirubin oxidase was screened from a soil sample in China. Pure bilirubin oxidase was obtained from the culture filtrate employing the QAE sephadex A50 column chromategraphy. Its specific activity was 207.65 U/A280 nm, total yield was 22.3%. The purified bilirubin oxidase exhibited characteristic UV-spectrum with a maximun at 278 nm and was homogeneous on disc gel electrophoresis.The molecular weight of the enzyme was estimated to be approximately 52000. The enzyme rapiday oxidazed bilirubin to biliverdin and farther to a pale purple pigment, the optimum temperature and pH for the enzyme reaction was about 40℃and pH 7.0 respectively.