双拷贝aveC基因对阿维菌素产量的影响  

Influence of doublecopy genes of aveC on avermectin production in Streptomyces avermitilis

作　　者：陈光[1] 张怡轩[1] 何建勇[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2008年第8期667-673,共7页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省企业博士后项目(辽博函2006-4);中国博士后项目(20070410356)

摘　　要：目的通过在阿维链霉菌染色体上增加aveC基因的拷贝数,提高阿维菌素"1"组分的产量。方法采用基因工程技术,在阿维链霉菌染色体DNA的bkdAB基因中插入一套aveC基因,通过同源双交换得到重组菌株,采用HPLC考察各组分的含量。结果第二套aveC基因正确地插入到基因组DNA的预定部位。然而,对重组菌株发酵产物的HPLC分析显示,阿维菌素"1"组分比例并没有明显的变化,但B1a效价却有显著提高。结论aveC基因产物的活性可能不是决定阿维菌素"1"组分比例的主要因素,但aveC基因产物的活性可能是阿维菌素生物合成的限制因素。Objective To enhance the production of avermectin component＂1＂ by increasing aveC copy in the chromosome of Streptornyces averrnitilis. Methods Second copy of aveC was inserted into bkdaB in the genomic DNA of S. avermitilis by gene manipulation;recombinant strains were obtained by homologous DNA double cross-over, and the content of each component was detected by HPLC. Results It was testified that the second aveC copy was inserted exactly into the expected site. However, HPLC analysis of fermentation products produced by recombinant strains showed that the ratio of component ＂1＂ to component ＂2＂ changed indistinctively, while Bla potency of avermectin by recombinant strains enhanced markedly. Conclusions It implies that the activity of aveC gene product may not be the main factor for the ratio of component＂1＂ to component ＂2＂, but its activity might be a restrictive factor in the biosynthesis of avermectin in S. averrnitilis.

关 键 词：阿维链霉菌 阿维菌素 aveC们基因 bkdAB基因 

分 类 号：Q939.9[生物学—微生物学]