阿维菌素衍生物CHC-B1的突变生物合成  

The Mutasynthetic Production of CHC-B1 from a Streptomyces avermitilis Mutant in the Presence of Cyclohexanecarboxylic Acid

作　　者：姜薇[1,2] 汪洋[2] 张晓琳[2] 郭伟群[2] 刘娣 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]国家粮食局科学研究院,北京100037

出　　处：《中国生物工程杂志》2009年第8期68-74,共7页China Biotechnology

基　　金：国家"863"计划(2006AA10A208);"十一五"国家科技支撑计划(2006BAD08B02)资助项目

摘　　要：利用成功构建的基因缺失载体pLJ04(pKC1139∷△bkdF+△bkdH)对阿维菌素(avermectin)高产菌阿维链霉菌(Streptomyces avermitilis)76-02-e的bkdFGH基因进行缺失,获得的bkdFGH缺失突变株经过摇瓶发酵和HPLC检测,发现该突变株完全丧失了产生阿维菌素的能力。2-甲基丁酸及异丁酸的前体添加试验表明,当有外源前体存在时,突变株又能恢复阿维菌素合成的能力。将该bkdFGH基因缺失突变株命名为S.avermitilisbkd76-3。环己羧酸(CHC)前体添加试验及HPLC检测发现存在4种产物,经LC/MS分析验证,其中两种产物分别为CHC-B1和CHC-A2。Gene deletion vector pLJ04（pKC1139：：△bkdF＋△6kdH）was used to disrupt bkdFGH in Streptomyces avermitilis 76-02-e, an industrial producer of anthelmintic avermectin. The disruptants were confirmed by PCR. Shake flask experiment and HPLC analysis showed that the mutant lost the ability to produce avermectins. As it is expected, the mutant, named S. avermitilis bkd76-3, could restore the ability of producing avermectins when the feeding of methylbutyric acid and isobutyric acid to its fermentations was carried out. The addition of cyclohexanecarboxylic acid （CHC） into fermentations of the S. avermitilis bkd76-3 allowed for production of four components, two of which was confirmed as CHC -B1 and CHC-A2 by LC/MS analysis, respectively.

关 键 词：阿维链霉菌 bkdFGH 基因缺失 多拉菌素 

分 类 号：Q819[生物学—生物工程]