Production of (2S)-sakuranetin from (2S)-naringenin in Escherichia coli by strengthening methylation process and cell resistance  

作　　者：Qiumeng Sun Song Gao Shiqin Yu Pu Zheng Jingwen Zhou 

机构地区：[1]Science Center for Future Foods,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [2]Key Laboratory of Industrial Biotechnology,Ministry of Education and School of Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [3]Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [4]Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology,Jiangnan University,Wuxi,214122,China

出　　处：《Synthetic and Systems Biotechnology》2022年第4期1117-1125,共9页合成和系统生物技术（英文）

基　　金：This work was supported by the National Key Research and Development Program of China(2019YFA0904800);the National Natural Science Foundation of China(21908078).

摘　　要：(2S)-Sakuranetin is a 7-O-methylflavonoid that has anticancer,antiviral,and antimicrobial activities.Methylation process is involved in biosynthesizing(2S)-sakuranetin from(2S)-naringenin,in which S-adenosylmethionine(SAM)serves as the methyl donor.In this study,after methyl donor and substrate inhibition were identified as limiting factors for(2S)-sakuranetin biosynthesis,an efficient(2S)-sakuranetin-producing strain was constructed by enhancing methyl donor supply and cell tolerance to(2S)-naringenin.Firstly,PfOMT3 from Perilla frutescens was selected as the optimal flavonoid 7-O-methyltransferase(F7-OMT)for the conversion of(2S)-naringenin to(2S)-sakuranetin.Then,the methylation process was upregulated by regulating pyridoxal 5′-phosphate(PLP)content,key enzymes in methionine synthesis pathway,and the availability of ATP.Furthermore,genes that can enhance cell resistance to(2S)-naringenin were identified from molecular chaperones and sRNAs.Finally,by optimizing the fermentation process,681.44 mg/L of(2S)-sakuranetin was obtained in 250-mL shake flasks.The titer of(2S)-sakuranetin reached 2642.38 mg/L in a 5-L bioreactor,which is the highest titer ever reported.This work demonstrates the importance of cofactor PLP in methylation process,and provides insights to biosynthesize other O-methylated flavonoids efficiently in E.coli.

关 键 词：(2S)-Sakuranetin Flavonoid 7-O-methyltransferases METHYLATION Cell tolerance Metabolic engineering 

分 类 号：O62[理学—有机化学]