食品与发酵工业2026，Vol.52Issue(7)：9-18,中插1-中插3,13.DOI:10.13995/j.cnki.11-1802/ts.042772
系统代谢工程重构大肠杆菌合成路径生产L-苯丙氨酸
Systematic metabolic engineering reconfigures biosynthetic pathway in Escherichia coli for L-phenylalanine production
摘要
Abstract
L-Phenylalanine,a critical intermediate in the food and pharmaceutical industries,requires enhanced biosynthetic efficien-cy to meet industrial demands.This study engineered the L-phenylalanine biosynthetic pathway in Escherichia coli W3110 through system-atic metabolic modifications.Initially,the regulatory genes lacI and tyrR were deleted to alleviate metabolic repression,while the feedback inhibition-resistant variant aroGfbr was overexpressed to reinforce the supply of the precursor DAHP.Subsequently,inactivation of csrA and poxB optimized carbon flux distribution,achieving simultaneous improvements in biomass accumulation and substrate utilization efficiency.Further metabolic optimization involved chromosomal integration of the feedback-resistant pheAfbr,ilvE,and aromatic biosynthesis module genes(aroB,aroD,aroK,aroC),coupled with deletion of the competitive pathway gene ydiB,to reconstruct a streamlined central meta-bolic network.Finally,integration of the transporter gene yddG and deletion of aroP enhanced product secretion,yielding the engineered strain M13.Quantitative reverse transcription polymerase chain reaction(RT-PCR)analysis confirmed the enhanced mRNA abundance of key genes post-engineering.In a 5 L fed-batch fermentation system,strain M13 produced 62.7 g/L L-phenylalanine with a glucose-to-product conversion yield of 22.3％after 60 h.This study elucidated the mechanistic impact of sequential metabolic engineering on L-phen-ylalanine biosynthesis,with transcriptional profiling revealing the regulatory effects of genetic modifications on metabolic flux distribution.These findings provide critical data for overcoming bottlenecks in L-phenylalanine biosynthesis and establish a methodological framework for developing high-yield microbial platforms for aromatic amino acids and their derivatives.关键词
大肠杆菌/L-苯丙氨酸/路径改造/代谢改造/逆转录聚合酶链式反应Key words
Escherichia coli/L-phenylalanine/engineering the synthesis pathway/metabolic modification/reverse transcription polymerase chain reaction(RT-PCR)引用本文复制引用
张颖,杨凤玉,廖雅芯,饶志明,徐美娟..系统代谢工程重构大肠杆菌合成路径生产L-苯丙氨酸[J].食品与发酵工业,2026,52(7):9-18,中插1-中插3,13.基金项目
国家重点研发计划项目(2023YFD1300700) （2023YFD1300700）
国家自然科学基金项目(32270036) （32270036）
中央高校基本科研业务费专项资金(JUS-RP221012,JUSRP622022) （JUS-RP221012,JUSRP622022）
