食品与发酵工业2024，Vol.50Issue(24)：76-83,8.DOI:10.13995/j.cnki.11-1802/ts.038642
γ-谷氨酰转肽酶在枯草芽孢杆菌中高效表达及发酵培养基优化
Efficient expression of γ-glutamyl transpeptidase in Bacillus subtilis and optimization of enzyme-producing culture medium
摘要
Abstract
γGlutamyl transpeptidase is an enzyme that can catalyze glutamine to generate a y-glutamyl-enzyme complex.Then the complex can be nucleophilically substituted by an acceptor substrate such as an amino acid or a short peptide,resulting in a transpeptida-tion reaction that forms a γ-glutamyl peptide.This enzyme has significant application value in the fields of food and medicine.In this re-search,Bacillus subtilis SCK6 was used as the expression host,based on 173 signal peptides of Bacillus subtilis,a signal peptide screening library was constructed,and the optimal signal peptide(SPcotC)was identified.Promoter screening was then carried out to obtain the opti-mal promoter(PaprE).Finally,with the optimal signal peptide and prompter,a recombinant strain BS-gts3 was constructed which could express and secrete γ-glutamyl transpeptidase.The maximum extracellular enzyme activity reached 5.04 U/mL in shake flask fermenta-tion.The optimal medium was further determined through one-way and response surface tests.The optimal composition of the medium was 31.73 g/L of lactose,50.26 g/L of FM802,1.24 g/L of MgSO4,1.25 g/L of KH2PO4,and 0.75 g/L of K2HPO4.The culture was then scaled up in a 5 L fermenter,and the maximum extracellular enzyme activity reached 28.26 U/mL after 38 h fermentation,which was the highest reported level.This study provides a foundation for the development and application of γ-glutamyl transpeptidase.关键词
枯草芽孢杆菌/γ-谷氨酰转肽酶/启动子/信号肽/重组表达Key words
Bacillus subtilis/γ-glutamyl transpeptidase/promoter/signal peptide/recombined expression引用本文复制引用
张祖政,毛泽敬,余华顺,张彦,杨潇,龚大春,郑贤良..γ-谷氨酰转肽酶在枯草芽孢杆菌中高效表达及发酵培养基优化[J].食品与发酵工业,2024,50(24):76-83,8.基金项目
国家重点研发计划资助(2021YFC2100300) （2021YFC2100300）
山东省重大科技创新工程项目(2022CXGC020712) （2022CXGC020712）
