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本文对烟草赤星病菌Alternaria longipes产生的角质酶进行了纯化及性质测定。在含有苹果角质(诱导物)的液体培养基中培养A.longipes,第8 d达到产酶高峰。发酵液经硫酸铵沉淀、DEAE(diethylaminoethyl)Sepharose、Sephacryl S-100 HR和Phenyl-Sepharose 6 fast flow(high sub)进行纯化。通过SDS-PAGE测得蛋白分子量为59.5 kDa。该酶的最适温度为35℃,最适pH为9.0,在温度20℃～40℃及pH 4.0～10.0范围内,酶活较稳定。一些金属离子如Na+、Cu2+和Mn2+可以部分抑制角质酶活性。
In this paper, the cutinase produced by Alternaria longipes was purified and characterized. A. longipes were cultured in liquid medium containing keratin (inducer), and reached peak on the 8th day. The fermentation broth was purified by ammonium sulfate precipitation, DEAE (diethylaminoethyl) Sepharose, Sephacryl S-100 HR and Phenyl-Sepharose 6 fast flow (high sub). The molecular weight of the protein was 59.5 kDa as measured by SDS-PAGE. The optimum temperature of the enzyme is 35 ℃, the optimum pH is 9.0, the enzyme activity is stable at the temperature of 20 ℃ ~ 40 ℃ and the pH value of 4.0 ~ 10.0. Some metal ions such as Na +, Cu2 + and Mn2 + can partially inhibit the cutinase activity.