朱黄青霉α-1,6-葡聚糖酶在毕赤酵母GS115中的异源表达  

Heteroexpression of α-1,6-Dextranase from Penicillium minioluteum in Pichia pastoris

作　　者：王赟[1] 章晓庆[2] 王风清[1] 

机构地区：[1]华东理工大学鲁华生物技术研究所,上海200237 [2]宁夏夏盛实业集团有限公司,上海200237

出　　处：《食品与药品》2014年第2期77-80,共4页Food and Drug

摘　　要：目的构建毕赤酵母基因工程菌,以实现朱黄青霉α-1,6-葡聚糖酶的异源高效表达。方法人工合成朱黄青霉HI-4的α-1,6-葡聚糖酶基因ZHdex,克隆到毕赤酵母分泌型表达载体pPIC9K,电转入毕赤酵母GS115。甲醇诱导目的蛋白表达。结果成功在毕赤酵母GS115中对ZHdex基因进行表达,SDS-PAGE表明重组毕赤酵母在66×103附近有目的蛋白表达,与理论值一致。在摇瓶水平,甲醇诱导培养120 h后,α-1,6-葡聚糖酶的最高酶活达28.79 U/mL,蛋白质浓度为0.56 mg/mL。结论获得一株重组毕赤酵母GS115-ZHdex,其产物具备α-1,6-葡聚糖酶活性,成功地实现朱黄青霉α-1,6-葡聚糖酶的异源高效表达。Objective To construct the recombinant Pichia pastoris to heteroexpress the α-1,6-dextranase from Penicillium minioluteum. Methods α-1,6-Dextranase gene （ZHdex） of Penicillium minioluteum was synthesized and cloned into vector pPIC9K, then transformed into the expression host P. pastoris GS115 via electroporation. Methanol was used for protein expression. Results ZHdex was successfully expressed, and the Mr of recombinantα-1,6-dextranase was estimated to be 66×10^3, which is consistent with the theoretical value. In shaking flasks, the highest α-1,6-dextranase activity was determined to be 28.79 U/mL with a protein concentration of 0.56 mg/mL after 120 h induction with methanol. Conclusion A recombinant P. pastoris GS115-ZHdex can be obtained and its product can possessα-1,6-dextranase activity. Theα-1,6-dextranase from Penicillium minioluteum can be successfully heteroexpressed.

关 键 词：朱黄青霉 α-1 6-葡聚糖酶 异源表达 毕赤酵母 

分 类 号：R969[医药卫生—药理学]