杀念菌素/FR-008生物合成途径中转运基因fscTⅠ与fscTII的功能  

Function of transporter genes fscTⅠ and fscTⅡ in the biosynthetic cluster of candicidin/FR-008

作　　者：雷璇[1] 孔令新[1] 张晨[1] 由德林[1] 邓子新[1] 

机构地区：[1]微生物代谢国家重点实验室,上海交通大学生命科学技术学院,上海200030

出　　处：《微生物学报》2012年第12期1458-1466,共9页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31170085);国家"973项目"--国家重点基础研究发展计划(2012CB721004)~~

摘　　要：【目的】分析杀念菌素/FR-008生物合成途径中转运基因fscTⅠ和fscTⅡ的功能。【方法】构建转运基因fscTⅠ和fscTⅡ的敲除质粒pJTU4137,并通过接合转移和同源重组双交换的方法得到转运基因缺失突变株。转运基因fscTⅠ和fscTII也被克隆到高拷贝质粒pJTU1278上用于在链霉菌FR-008(Streptomyces sp.FR-008)的衍生菌株ZYJ-6中进行转运蛋白的过量表达。【结果】获得了转运蛋白缺失的双交换突变株LX10,发酵结果显示该突变株不再产生杀念菌素及其衍生物;过量表达转运蛋白的基因工程菌株LX11,其杀念菌素的产量约是对照菌株的1.5倍。【结论】体内遗传实验进一步证实FR-008生物合成途径中的fscTⅠ和fscTⅡ是ATP依赖的ABC转运基因,fscTⅠ与fscTⅡ的过量表达增加了杀念菌素的产量,为利用此方法提高其它多烯类抗生素的产量提供了例证。[Objective]To investigate function of transporter genes fscTⅠ and fscTⅡ in the biosynthetic gene cluster of candicidin/FR-008.[Methods]We constructed a plasmid pJTU4137 for disruption of transporter genes fscTⅠ and fscTⅡ by conjugation and homologous recombinant.The transporter genes were also PCR amplified and cloned into the high-copy plasmid pJTU1278 for overexpression in strain ZYJ-6 derived from Streptomyces sp.FR-008.[Results]The disruption mutant LX10 was unable to produce candicidin and its analogues.Overexpression of FscTI and FscTⅡ in ZYJ-6 caused a 1.5-fold increase in FR-008-Ⅲ production compared with the control.[Conclusion]We confirmed that fscTⅠ and fscTⅡ are function as ATP dependent ATP binding cassetle（ABC） transporters in the biosynthetic gene cluster of FR-008.Furthermore,a positive example was provided for improving antibiotic production in other polyene producing strains based on the results that overexpression of fscTⅠ and fscTⅠ increased candicidin production.

关 键 词：多烯类抗生素 ABC转运蛋白 基因敲除 基因过量表达 

分 类 号：Q933[生物学—微生物学]