食品与发酵工业2023，Vol.49Issue(23)：16-24,9.DOI:10.13995/j.cnki.11-1802/ts.035600
代谢改造大肠杆菌合成8-异戊烯基山奈酚
Metabolic engineering of Escherichia coli for 8-prenylkaempferol synthesis
摘要
Abstract
8-Prenylkaempferol(8-pk)is a flavonoid with excellent anti-inflammatory and anticancer pharmacological activities.At present,the biosynthesis of 8-pk is mainly obtained from kaempferol(Kae)by the catalysis of plant-derived prenyltransferases(PTs).However,plant-derived PTs contain signal peptides,which greatly hinder their heterologous expression and efficient catalysis in pro-karyotes.In addition,the insufficient supply of the precursor molecule dimethylallyl pyrophosphate(DMAPP)in the catalytic pathway se-verely limits the biosynthesis yield of 8-pk.In this study,Escherichia coli was used as a chassis cell,further enhancing the supply of pre-cursor DMAPP and truncating the signal peptide of prenyltransferase for 8-pk efficient synthesis.Firstly,epimedium derived prenyltrans-ferase(EkF8DT)was expressed in E.coli BL21(DE3)and IUP pathway was introduced to increase the supply of DMAPP,with an 8-pk titer of 2.14 mg/L.Subsequently,the structure of EkF8DT was predicted by AlphaFold2.Subsequently,the structure of EkF8DT was predicted by AlphaFold2 and the highest 8-pk yield of 6.46 mg/L was achieved when the N-end truncation of EkF8DT was truncated by 60 amino acids.By fusing the protein tag to the N-end truncation of EkF8DT and optimizing fermentation conditions a titer of 24.28 mg/L was achieved.Finally,8-pk titer of 44.33 mg/L was achieved in a 5 L fermenter,which is the higher level reported.This study provides an effective strategy for 8-pk biosynthesis.关键词
8-异戊烯基山奈酚/异戊烯基转移酶/异戊烯醇利用途径/信号肽截短/蛋白质标签Key words
8-prenylkaempferol/prenyltransferases/IUP pathway/signal peptide truncation/protein tags引用本文复制引用
赵婉莹,周景文,侯颖..代谢改造大肠杆菌合成8-异戊烯基山奈酚[J].食品与发酵工业,2023,49(23):16-24,9.基金项目
国家自然科学基金创新研究群体项目(32021005) （32021005）
天津市研究生科研创新项目资助(2022SKYZ092,2022SKYZ094,2022SKYZ099) （2022SKYZ092,2022SKYZ094,2022SKYZ099）
