提高耐热链霉菌转化泰乐菌素能力的分子改造与育种  

Molecular Manipulation and Breeding of Streptomyces thermotolerans to Increase the Bioconversion Ability of Tylosin

作　　者：何群香[1] 闵勇[2] 王华丽 梁运祥[1] 吕和平[4] 邢仁昌[4] 郑应华[1,4] 

机构地区：[1]华中农业大学农业微生物国家重点实验室,武汉430070 [2]湖北省农业科学院,武汉430064 [3]中牧股份兽药研究所,北京100091 [4]北京生物医药研究所,北京100091

出　　处：《农业生物技术学报》2008年第6期1012-1018,共7页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)(No.2006AA10A208)项目资助

摘　　要：BIB0830是将泰乐菌素转化为乙酰异戊酰泰乐菌素(AIV)的耐热链霉菌(Streptomyces thermotolerans)工业菌株。以pIJ8600为载体分别构建了acyB1-B2基因整合型表达载体pBIB425和带红霉素突变启动子ermEp*的红霉素抗性ermE基因整合型表达载体pBIB304。通过大肠杆菌(Escherichia coli)-链霉菌属间接合转移分别将pBIB425、pBIB304和pBIB308(耐热链霉菌的nsdA基因中断载体)导入工业菌株BIB0830,抗性筛选分别得到接合子BIB425、BIB304和BIB308。经PCR验证均为阳性克隆。摇瓶发酵结果表明,与出发菌株BIB0830相比,BIB425和BIB308泰乐菌素转化率分别提高了14%和22%,而BIB304则无明显影响。The industrial Streptomyces thermotolerans strain BIB0830 can bioconverse tylosin to acetyl-isovaleryl tylosin （AIV）. Recombinant plasmids pBIB425 containing structural gene acyB1-B2 and pBIB304 containing structural gene ennE with up-mutant promoter of ermE （ermEp^＊）, were derived from pLJ8600, The recombinant strain BIB425, BIB304 and BIB308 were obtained by introducing pBIB425, pBIB304 and pBIB308 containing disruption cassette of nsdA gene respectively into the industrial strain BIB0830 through intergeneric conjugal transfer. After PCR analysis, they were confirmed to be the positive clones. Compared with the starting strain BIB0830, at the level of flask, the bioconversion abilities of tylosin to acetyl-isovaleryl tylosin （AIV） of recombinant BIB425 and BIB308 increased 14% and 22%, respectively, but BIB304 was not affected.

关 键 词：acyB1-B2基因 ermE基因 nsdA基因 乙酰异戊酰泰乐菌素 耐热链霉菌 

分 类 号：S188[农业科学—农业基础科学]