CAJ | 학술논문

本文对烟草赤星病菌Alternuria 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 S100 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~+、Cu~(2+)和Mn~(2+)以部分抑制角质酶活性.
본문대연초적성병균Alternuria longipes산생적각질매진행료순화급성질측정.재함유평과각질(유도물)적액체배양기중배양A.longipes,제8 d체도산매고봉.발효액경류산안침정、DEAE(diethylaminoethyl)Sepharose、Sephacryl S100 HR화Phenyl-Sepharose 6 fast flow(high sub)진행순화.통과SDS-PAGE측득단백분자량위59.5 kDa.해매적최괄온도위35℃,최괄pH위9.0,재온도20℃～40℃급pH 4.0～10.0범위내,매활교은정.일사금속리자여Na~+、Cu~(2+)화Mn~(2+)이부분억제각질매활성.
Cutinase from Alternaria longipes was purified and characterized. A. longipes was grown in a liquid culture medium containing apple cutin as the inducer. After 8 d of growth, the cutinase was purified through ammonium sulphate precipitation, DEAE (diethylaminoethyl) Sepharose, Sephacryl S-100 HR, and Phenyl-Sepharose 6 fast flow (high sub). A single band was observed by SDS-PAGE, corresponding to a molecular weight of 59.5 kDa. Optimum temperature and pH for the cutinase activity were 35t and pH 9.0, respectively. The enzyme activity was stable within 201 -40℃ and pH 4. 0 -10. 0. Na~+ , Cu~(2+) and Mn~(2+) could partially inhibit the cutinase activity.