食品与发酵工业2025，Vol.51Issue(24)：1-8,8.DOI:10.13995/j.cnki.11-1802/ts.042318
重组溶菌酶在谷氨酸棒杆菌中的分泌表达
Secretion of recombinant lysozyme in Corynebacterium glutamicum
摘要
Abstract
Lysozyme,also known as cell wall hydrolase or peptidoglycan hydrolase,is a natural antimicrobial enzyme with broad-spectrum antibacterial activity and has been widely used in food preservation.Corynebacterium glutamicum,a food-safe strain,shows sig-nificant potential for lysozyme production.This study aimed to achieve the secretion and expression of recombinant lysozyme in C.glutami-cum CGMCC1.15647.First,the expression of lysozymes from different sources in C.glutamicum was analyzed,and the lysozyme with op-timal expression efficiency was selected as the target protein.The pGX19 vector was employed for secretory expression.Subsequently,multi-level optimizations were performed,including signal peptide screening,promoter and ribosome binding site(RBS)combinations,and enhancement of the transport system.Results showed that the Tat pathway signal peptide CgR0949 significantly improved recombinant lysozyme expression.After promoter and RBS optimization,the expression level increased by 2.3-fold.Overexpression of the Tat transport system from diverse sources further enhanced recombinant lysozyme production by 1.5-fold.Through these optimizations,the shake-flask fermentation yield of recombinant lysozyme reached 0.2 g/L,representing an 8.3-fold improvement over the initial strain.Finally,the re-combinant protein was purified,and its enzymatic properties were characterized,revealing optimal activity at pH 7.5 and 35 ℃.This stud-y is the first to achieve secretory expression of recombinant lysozyme in C.glutamicum.By integrating signal peptide screening,transcrip-tion-translation element optimization,and transport system reinforcement,it provides a novel strategy for efficient lysozyme production.关键词
谷氨酸棒杆菌/溶菌酶/表达元件优化/分泌表达/信号肽优化Key words
Corynebacterium glutamicum/lysozyme/expression element optimization/secretory expression/signal peptide optimization引用本文复制引用
田明科,邹宇,武小月,白仲虎,李康,刘秀霞..重组溶菌酶在谷氨酸棒杆菌中的分泌表达[J].食品与发酵工业,2025,51(24):1-8,8.基金项目
国家自然科学基金项目(22378167) （22378167）
