3-O-乙酰转移酶的表达及其酶活特性研究

Expression and Characterization of 3-O-Acetyltransferase

将3个不同来源的3-O-乙酰转移酶基因转入大肠杆菌表达宿主,成功实现3种3-O-乙酰转移酶的异源表达,并通过诱导条件优化提高了可溶蛋白表达量。酶活特性研究结果显示,3种不同来源的乙酰转移酶在不同温度及pH条件下活性有着显著差异,其中Fo1TRI201最适温度及pH分别为40℃及pH=5.0,有着更强的温度及pH耐受性。最后在工业物料中考察了粗酶液的脱毒效果,通过向玉米浸泡液中添加1%的Fo1TRI201粗酶液,在40℃、pH=5.0条件下24 h可将玉米浸泡液中的DON从2000μg·L^(-1)以上降解至500μg·L^(-1)以下,降解率最高达82.64%,为呕吐毒素脱毒产品和技术开发提供了选择和参考,具有一定的商业应用价值。

Three 3-O-acetyltransferase genes from different strains were transferred into the expression host of E.coli,and were successfully expressed.The expression of soluble proteins was improved by optimizing the induction conditions.The enzyme characteristics study showed that the activity of acetyltransferase from three different strains was significantly different under different temperature and pH conditions,among which the optimum temperature and pH of Fo1TRI201 were 40℃and pH=5.0,respectively,with stronger temperature and pH tolerance.Finally,the detoxification effect of the crude enzyme solution was investigated in industrial materials.By adding 1%Fo1TRI201 crude enzyme solution to the corn soaking solution,the DON in the corn soaking solution was degraded from more than 2000μg·L^(-1)to less than 500μg·L^(-1)under the condition of 40℃and pH=5.0 in 24 h,with the highest degradation rate of 82.64%.Our study provides a choice and reference for the development of detoxification products and technologies for DON,and has certain commercial application value.