γ-环糊精糖基转移酶在毕赤酵母中高效表达  

High-level expression of codon optimized γ-cyclodextrin glycosyltransferase by Pichia pastoris

作　　者：张燕[1] 李梦腊 张建国[1] ZHANG Yan LI Mengla ZHANG Jianguo(Institute of Food Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, P.R. Chin)

机构地区：[1]上海理工大学食品科学与工程研究所,上海200093

出　　处：《工业微生物》2017年第1期24-30,共7页Industrial Microbiology

基　　金：国家自然科学基金(No.21306112);上海市自然科学基金资助项目(No.13ZR1429100);上海高校青年教师培养资助计划(No.slg14037);教育部留学回国人员科研启动基金

摘　　要：本论文通过密码子优化、表达质粒的比较、发酵条件优化,将来源于Bacillus pseudalcaliphilus的环糊精糖基转移酶基因在毕赤酵母中高效表达。在诱导初始p H 6.0、发酵温度28℃、接种量6%、甲醇添加量1%时,γ环化活力最终达到了83.65 U/m L,比活力296.49 U/mg。通过硫酸铵分步沉淀法即可有效将上清粗蛋白盐析制备,得到68.28%γ-环糊精糖基转移酶。In this work, the gene sequence of y-cyclodextrin glycosyltransferase from Bacillus pseudalcaliphilus was optimized, the expression of Pichia pastoris was selected and its cultivation parameters were optimized for high level expression by Pichia pastoris. Under the conditions of pH 6.0, 28 ℃ , 6.0% inoeulum size and 1.0% methanol addition per 24 h, γ-cyelodextrin glycosyltransferase yield reached 83.65 U/mL, which was 8.7 folds of previous results. Finally, 68.28% γ-cyclodextrin glycosyltransferase was recovered through ammonium sulfate precipitation.

关 键 词：γ-环糊精糖基转移酶 密码子优化 毕赤酵母 发酵优化 

分 类 号：Q78[生物学—分子生物学]