透明颤菌血红蛋白基因vgb和腈水解酶基因bxn在毕赤酵母中的表达研究  

Expression of Vitreoscilla Hemoglobin and Nitrilase in the Yeast Pichia pastoris

作　　者：王清路[1] 张锐[1] 倪万潮[1] 陈毓荃[2] 郭三堆[3] 

机构地区：[1]江苏省农业科学院遗传生理研究所,南京210014 [2]西北农林科技大学生命科学学院,杨凌712100 [3]中国农业科学院生物技术研究所,北京100081

出　　处：《生物工程学报》2004年第5期730-735,共6页Chinese Journal of Biotechnology

基　　金：江苏省农业科学院博士后基金 (No .0 110 2 0 1)~~

摘　　要：为获得高效表达外源蛋白的Pichiapastoris菌株而设计了重组质粒pPIC9K_vgbbxn ,其中透明颤菌血红蛋白基因vgb胞内表达以提高菌体的发酵密度 ,腈水解酶基因bxn分泌表达。转化GS115菌株后 ,通过PCR、SDS_PAGE检测证实两基因已经整合进酵母基因组且能高效表达 ,以及用准确的蛋白活性测定方法成功地检测到二者所表达的产物均具有正常的活性。摇瓶发酵实验证明 。The expression of the vgb gene in vivo could improve the fermentation density and then contribute the extracellular secretion of the product of bxn gene. Constructed the recombination plasmid pPIC9K-vgbbxn and transformed into Pichia pastoris GS115 The results of PCR and SDS-PAGE indicate that the vgb gene and bxn gene had integrated into the genome of Pichia pastoris GS115 and expressed in efficient level. Also, the protein activity of their products had been verified respectively. Shake flask fermentation experiments showed that the presence of VHb in yeast Pichia pastoris efficiently enhanced cell growth and secretive expression of bxn gene under hypoxic habitats.

关 键 词：毕赤酵母 分泌型表达载体 腈水解酶 透明颤菌血红蛋白 

分 类 号：Q786[生物学—分子生物学]