刺糖多孢菌高产菌株转录组及其多杀菌素合成代谢调控研究  

Metabolic regulation of Saccharopolyspora spinosa based on transcriptomic analysis

作　　者：王欣荣[1] 唐智超 曾茂 吕正 翟龙飞[1] 张新宜[1] 刘超兰[1] 褚以文[1] 王克华 黄挺 Wang Xin-rong;Tang Zhi-chao;Zeng Mao;LüZheng;Zhai Long-fei;Zhang Xin-yi;Liu Chao-lan;Chu Yi-wen;Wang Ke-hua;Huang Ting(Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province,School of pharmacy,Chengdu University,Chengdu 610052;North China Pharmaceutical Group Aino Co.,Ltd,Shijiazhuang 052165)

机构地区：[1]成都大学药学院,抗生素研究与再评价四川省重点实验室,成都610106 [2]华北制药集团爱诺有限责任公司,石家庄052165

出　　处：《中国抗生素杂志》2022年第6期561-568,共8页Chinese Journal of Antibiotics

基　　金：“重大新药创制”化学结构导向的分子设计育种体系建设(No.2018ZX09711001-007-006);四川省科技计划项目(No.2020YJ0276)。

摘　　要：目的 通过对高产多杀菌素的刺糖多孢菌进行转录组分析,挖掘多杀菌素合成相关代谢通路的差异基因,并在此基础上结合发酵优化进一步提高多杀菌素产量。方法 利用RNA-seq技术对高产株SS-168及低产野生菌株SS-WT进行比较转录组分析,通过荧光定量PCR验证差异基因表达,再通过发酵培养基优化考察高产菌株的发酵水平。结果 转录组分析表明,与低产菌株相比高产株中有1341个差异表达基因,GO注释表明DEGs主要参与氧化还原生物过程和脂质代谢及辅因子结合和辅酶结合,主要分布在甘氨酸/丝氨酸/苏氨酸代谢、脂肪酸降解等通路。荧光定量PCR结果与转录组数据一致性达到100%。采用分别含有不同浓度的甘氨酸/丝氨酸/苏氨酸的发酵培养进行筛选,多杀菌素发酵水平显著提高。结论 本研究通过新颖的转录组学方法获得了刺糖多孢菌高产菌株的差异基因,并对其高产机制和发酵优化进行了初步分析,为改良刺糖多孢菌菌株和优化发酵工艺提供重要的理论基础和参考依据。Objective Transcriptome analysis of Saccharopolyspora spinosa(S. spinosa) with high yield of spinosad was carried out to explore the differentially expressed genes(DEGs) in metabolic pathways related to spinosad synthesis, and combined with fermentation optimization to further improve the spinosad production.Methods RNA-seq technology was used for comparative transcriptome analysis of high-yield strain SS-168 and low-yield strain SS-WT. The expression of differential genes was verified by quantitative PCR. The fermentation level of high-yield strain was investigated by optimization of fermentation medium. Results Transcriptome analysis showed that 1341 DEGs were detected in SS-168 strain compared with SS-WT strain. GO annotation showed that DEGs were mainly involved in redox biological processes, lipid metabolism, cofactor binding and coenzyme binding,and mainly distributed in glycine/serine/threonine metabolism, fatty acid degradation and other pathways. In addition,the mRNA levels of 7 genes in the glycine/serine/threonine metabolism pathway were significantly up-regulated in the SS-168 strain compared to the SS-WT strain. The fermentation production of spinosad was significantly improved by adding appropriate concentrations of glycine/serine/threonine. Conclusion In this study, the DEGs of SS-168 strain were obtained by novel transcriptomics method, and the fermentation optimization were preliminarily analyzed, which provided an important strategy for improving S. spinosa strain and optimizing fermentation process.

关 键 词：多杀菌素 刺糖多孢菌 转录组 代谢调控 

分 类 号：R978.1[医药卫生—药品]