第 33 卷第 2 期 Vol. 33 No. 2 2003 年 4 月 Apr 2003

所属栏目：其它 用毕氏酵母表达蛇岛蝮蛇毒类凝血酶发酵条件的优化

龙姝; 黄永华; 杨青(长沙电力学院侯家塘校区; 大连理工大学化工学院; 湖南长沙; 410015; 116012)

摘　要：为了提高大连蛇岛蝮蛇毒类凝血酶的表达量 ,利用 5 0 0ml摇瓶培养体系研究了在不同培养基、pH值、发酵时间等培养条件下分泌蛋白的浓度及活性 ,使发酵条件得到优化 :丰富培养基比营养缺陷型培养基更利于目的蛋白的分泌表达 ,发酵优惠条件 pH值为 6 0 ,时间为 36h ,发酵液上清能检测到靶酶的酰胺水解活性。类凝血酶的表达量约为 4mg/L。

关键词：蛇岛蝮蛇; 类凝血酶; 毕氏酵母; 分泌表达; 优化

中图分类号：q815　　文献标识码：A　　文章编号：1009-9212(2003)02-0042-03

Optimization of Fermentation Conditions for Express of Gloshedobin in Yeast Pichia Pastoris

LONG Shu 1; HUANG Yonghua 1; YANG Qing 2.（1.Dept.of Core Curriculm; Changsha University of Electric Power; Changsha; 410015; China; 2.Department of Bioengineering School of Chemical Engineering Dalian University of Technology; Dalian 116012; China）

Abstract：To increase the expression yield of Gloshedobin by Pichia pastrois,the recombinant thrombin-like enzyme was expressed ith 500ml shaking flask.By studying the concentration and activity of secreted protein under different conditions:components of medium,pH,and the time of fermentation,the optimal fermentation conditions were obtained:rich medium is more beneficial for secretion expression than minimal medium,the optimal pH 6.0, and the optimal fermentation time 36 hours.The amidolytic actvity can be detected...

Key words：gloydius shedaoensis; thrombin-like enzyme; pichia pastoris; expression; optimization

基金项目：辽宁省科学技术基金资助项目(NO.99305001)

收稿日期：2003-01-20

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