卡西霉素生物合成途径中抗性基因calT的功能  

The function of the resistance gene calT in the biosynthetic pathway of calcimycin

作　　者：苟丽霞 解子晗 张文丽 宋子涵 汪志军[3] 韩铁生 Gou Li-Xia;Xie Zi-han;Zhang Wen-li;Song Zi-han;Wang Zhi-jun;Han Tie-sheng(School of Life Sciences,North China University of Science and Technology,Tangshan 063210;Hebei Province Key Laboratory of Occupational Health and Safety for Coal Industry,School of Public Health,North China University of Science and Technology,Tangshan 063210;State Key Laboratory of Microbial Metabolism,School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240)

机构地区：[1]华北理工大学生命科学学院,唐山063210 [2]华北理工大学公共卫生学院,河北省煤矿卫生与安全重点实验室,唐山063210 [3]上海交通大学生命科学技术学院微生物代谢国家重点实验室,上海200240

出　　处：《中国抗生素杂志》2023年第6期643-648,共6页Chinese Journal of Antibiotics

基　　金：国家自然科学基金项目(No.NSFC 31700083);河北省自然科学基金项目(No.H2020209001)。

摘　　要：目的研究卡西霉素产生菌教酒链霉菌NRRL 3882中编码MmpL家族同源蛋白的抗性基因calT的功能。方法利用λ-RED同源重组方法构建calT基因敲除质粒,通过接合转移和同源重组的方法得到双交换的calT基因敲除的突变株。通过HPLC分析突变株的代谢产物,并对突变株及野生菌株的细胞内及细胞外卡西霉素的分布进行分析。结果突变株GLX30(ΔcalT)中卡西霉素的产量与野生菌株相比显著降低,且胞内卡西霉素的占比与野生菌株相比增加约3倍,说明calT与卡西霉素的转运功能相关。结论CalT为转运蛋白,负责将卡西霉素转运至胞外以实现教酒链霉菌对卡西霉素的抗性。Objective To study the function of the resistance gene calT encoding MmpL family homologous protein in Streptomyces chartreusis NRRL 3882.Methods The calT gene knockout plasmid was constructed byλ-RED homologous recombination method,and the double-crossover calT gene knockout mutant was obtained by the methods of conjugation and homologous recombination.The metabolites of the mutant strains were analyzed by HPLC,and the distribution of intracellular and extracellular calcimycin in the mutant and wild strains was analyzed.Results Compared with the wild strain,the production of calcimycin in the mutant GLX30(ΔcalT)was significantly lower than that in the wild strain,and the proportion of intracellular calcimycin increased by about 3 times compared with the wild strain,indicating that calT and the wild strain related to the transport function of the hormone.Conclusion CalT is a transporter protein,which is responsible for transporting calcimycin to the outside of the cell to achieve the resistance of Streptomyces oleifera to calcimycin.

关 键 词：卡西霉素 calT MmpL家族 生物合成 

分 类 号：Q933[生物学—微生物学]