农业生物技术学报2025，Vol.33Issue(1)：179-189,11.DOI:10.3969/j.issn.1674-7968.2025.01.016
核糖体蛋白RpsL调控须糖多孢菌中丁烯基多杀菌素的合成
Ribosomal Protein RpsL Regulates Butenyl-spinosyn Synthesis in Saccharopolyspora pogona
摘要
Abstract
Butenyl-spinosyn,a secondary metabolite produced by fermentation of Saccharopolyspora pogona,is a green bio-insecticide.However,due to the low yield of fermentation and can not meet the needs of industrial production,the breeding of strains with high production performance of butenyl-spinosyn is the key to realize industrial production.In this study,ribosomal protein S12(RpsL)and RpsL mutant gene overexpression engineering strains Q8(rpsl)、W43(rpslK88R)、W44(rpslR86P)、W45(rpslK88E)和 W46(rpslK43N)were constructed were constructed using Saccharopolyspora pogona as the base cells.The effect of RpsL on the synthesis of butenyl-spinosyn was studied by shaking flask fermentation and comparison of RpsL protein structure.The results showed that the engineered strains Q8,W43,W44,W45,W46 increased the yield of butenyl-spinosyn to 275％,146％,111％,122％and reduced to 75％,respectively.The overexpression of rpsl and rpsslK88R genes increased the transcription level of the synthetic pathway genes,and thus significantly increased the yield of butenyl-spinosyn.At the same time,homologous modeling of the two was conducted,and it was found that RpsLK88R mutant structure had a larger deflection of the side chain compared with RpsL,which affected the binding effect to mRNA and thus reduced the ribosome translation efficiency,so its yield extraction effect was slightly lower than RpsL.This study provides a reference for protein engineering RpsL to improve the yield of butenyl-spinosyn.关键词
丁烯基多杀菌素/核糖体蛋白/须糖多孢菌/基因突变Key words
Butenyl-spinosyn/Ribosomal protein/Saccharopolyspora pogona/Gene mutation分类
农业科技引用本文复制引用
苏畅,王靖楠,郭超,徐周钦,李春,张根林,王超..核糖体蛋白RpsL调控须糖多孢菌中丁烯基多杀菌素的合成[J].农业生物技术学报,2025,33(1):179-189,11.基金项目
国家重点研发计划(2023YFA0914700) （2023YFA0914700）
国家自然科学青年基金(22108153) （22108153）
中央公益性科研院所专项课题(JY2311) （JY2311）
工业生物催化教育部重点实验室(清华大学)开放基金资助项目(2023003) （清华大学）
