Characterization of the positive SARP family regulator PieR for improving piericidin A1 production in Streptomyces piomogeues var.Hangzhouwanensis  

作　　者：Yan Li Lingxin Kong Jufang Shen Qing Wang Qian Liu Weinan Yang Zixin Deng Delin You 

机构地区：[1]State Key Laboratory of Microbial Metabolism,Joint International Research Laboratory of Metabolic and Developmental Sciences,School of Life Sciences&Biotechnology,Shanghai Jiao Tong University,Shanghai,China

出　　处：《Synthetic and Systems Biotechnology》2019年第1期16-24,共9页合成和系统生物技术（英文）

基　　金：the National Natural Science Foundation of China(31630002,31700029,31770038,31470183,21661140002 and 31170085);the Ministry of Science and Technology;Shanghai Pujiang Program from the Shanghai Municipal Council of Science and Technology(12PJD021);China Postdoctoral Science Foundation(2017M620151).

摘　　要：Piericidin A1,a member ofɑ-pyridone antibiotic,exhibits various biological activities such as antimicrobial,antifungal,and antitumor properties and possesses potent respiration-inhibitory activity against insects due to its competitive binding capacity to mitochondrial complex I.The biosynthetic pathway of piericidin A1 has been reported in Streptomyces piomogeues var.Hangzhouwanensis,while the regulatory mechanism remains poorly understood.In this study,a Streptomyces antibiotic regulatory protein(SARP)family transcriptional regulator PieR was characterized.Genetic disruption and complementation manipulations revealed that PieR positively regulated the production of piericidin A1.Moreover,the overexpression of pieR contributed to the improvement of piericidin A1 productivity.The real-time quantitative PCR(RT-qPCR)was carried out and the data showed that pieR stimulated the transcription of all the biosynthesis-related genes for piericidin A1.In order to explore the regulatory mechanism,electrophoresis mobility shift assays(EMSA)and DNase I footprinting experiments have been conducted.A protected region covering 50 nucleotides within the upstream region of pieR was identified and two 5-nt direct repeat sequences(5′-CCGGA-3′)in the protected region were found.These findings,taken together,set stage for transcriptional control engineering in the view of optimizing piericidin A1 production and thus provide a viable potent route for the construction of strains with high productivity.

关 键 词：ɑ-pyridone antibiotic Transcriptional regulation SARP family regulator Secondary metabolic STREPTOMYCES 

分 类 号：F42[经济管理—产业经济]