牛凝乳酶基因在毕赤酵母中的表达  

Expression of bovine chymosin in Pichia pastoris

作　　者：冷欢 张幸[1] 刘家书[1] 赵勇[1] 李谞[1] 江正兵[1] 宋慧婷[2] 

机构地区：[1]湖北省工业生物技术重点实验室(湖北大学),湖北武汉430062 [2]湖北大学资源环境学院,湖北武汉430062

出　　处：《湖北大学学报（自然科学版）》2014年第3期262-266,共5页Journal of Hubei University：Natural Science

基　　金：湖北省科技厅公益研究项目(2012DBA28001);武汉市科技计划项目(2013060501010165)资助

摘　　要：构建3株表达凝乳酶的重组毕赤酵母，通过甲醇诱导收集BMMY液体培养基上清后，通过ArimaK方法分别对其进行酶活测试并进行酶学性质分析，结果表明，GS115／pPICgK-chyl菌株诱导7d后，其酶活达到1.905 SU/mL重组凝乳酶的最适反应温度为45℃；最适反应pH为6；存pH4-7之间凝乳活性相对稳定；在50℃以下可以保持酶活，在55℃、处理50min后，丧欠60％的活性．Three recombinant Pichia pastoris expressing chymosin were constructed successfully in this experiment. Methanol was used to induce them, then culture supernatants BMMY were collected for further study. The enzyme activity was tested according to method of Arima K. The characterizations of chymosin from recombinant Pichia pastoris were also studied. Results showed that the optimal temperature and pH was 45 ℃ and 6.0 respectively. After GS115/pPIC9K chyl strain induced for 7 days, the enzyme activity of the supernatant reached 1. 905 SU/mL. The activity of recombinant chymosin was remained from pH 4. 0 to 7. 0 and below 50℃. When treated at 55 ℃ for ,50 minutes, the activity of recombinant chymosin was 40 percent of the original.

关 键 词：巴斯德毕赤酵母 牛凝乳酶 基因表达 凝乳酶性质 

分 类 号：Q784[生物学—分子生物学]