重组枯草芽孢杆菌的构建及对甾体的C_(1，2)位脱氢  

Researches on construction of recombinant Bacillus subtilis and C_(1,2)dehydrogenation of steroids

作　　者：李玉[1] 路福平[1] 王稳航[1] 杜连祥[1] 

机构地区：[1]天津市工业微生物重点实验室天津科技大学生物工程学院,天津300222

出　　处：《现代生物医学进展》2006年第9期15-17,23,共4页Progress in Modern Biomedicine

摘　　要：本文将简单节杆茵3-甾酮-1-脱氢酶基因克隆到分泌表达载体pWB980上,并转入枯草芽孢杆菌WB600中,得到重组芽孢杆菌菌株。重组芽孢杆菌表达出的目的蛋白的分子量为55KDa,分光光度法检测到胞内和胞外可溶性部分的酶活分别为110±0．5mU和15±0．6mU每毫克蛋白,对甾体底物4-AD的转化率为45．3%,比出发茵简单节杆菌提高了将近10倍。3 - ketostemid - △^1 - dehydrogenase gene from Arthrobaeter Simplex was cloned onto the expression vector pWB980 and then was transformed into Bacillus subtihsWB600. The molecular weight of the enzyme expressed by recombinant Bacillus subfilis was about 55KDa. The activity assayed by speetrophotometry of extracellular and intracellular soluble enzyme was 110 ± 0.5mU and 15 ± 0.6mU per milligram of protein respectively, and the transformation rate of androst - 4- ene - 3, 17 - dione was 45.3 %. Compared with Arthrobaeter Simplex, the transformation level of androst - 4 - ene - 3,17 - dione of Bacillus subfihs recombination cells was improved about ten-fold.

关 键 词：3-甾酮-1-脱氢酶 重组枯草芽孢杆菌 甾体转化 4-AD 

分 类 号：Q789[生物学—分子生物学]