食品与发酵工业2024，Vol.50Issue(11)：9-17,9.DOI:10.13995/j.cnki.11-1802/ts.038280
枯草芽孢杆菌高效分泌表达盒的构建和应用
Construction and application of high-efficiency secretory expression box for Bacillus subtilis
摘要
Abstract
The purpose of this work was to investigate the compatibility between different elements in the expression system of Bacillus subtilis,constructing an efficient protein secretion vector,which could be further applied to improve the extracellular expression level of chitosanase Csn21c.Firstly,12 plasmids containing different promoters(P43,Pshuttle09),ribosomal binding sites(RBS1,RBS5),and signal peptides(SnprB,SphoD,SymzC)were constructed,and the protein secretion levels of different element combinations were com-pared with three proteins(agarase AgWH50C,chitosanase Csn21c,and lipase Lip15)as indicator proteins,and the vectors pP435Y*K(P43-RBS5-SymzC)and pP095P*K(Pshuttle09-RBS5-SphoD)was superior to other vectors.By using vector pP435Y*K,the activity of agar AgWH50C enzyme increased by 184.25％,the activity of lipase Lip15 enzyme increased by 52.8％,and the activity of chitosanase Csn21c enzyme increased by 164.62％compared with that of the original vector(pP431N*K).Finally,by comparing five different media,it was determined that the TB medium was the most suitable fermentation medium for chitosanase Csn21c,and the highest enzyme activity of chitosanase Csn21c in the TB medium reached 21.14 U/mL,which was 128.31％higher than that of LB medium.关键词
枯草芽孢杆菌/分泌/启动子/核糖体结合位点/信号肽/壳聚糖酶Key words
Bacillus subtilis/secretion/promoter/ribosome binding site/signal peptide/chitosanase引用本文复制引用
韩瑞,初文琴,刘振..枯草芽孢杆菌高效分泌表达盒的构建和应用[J].食品与发酵工业,2024,50(11):9-17,9.基金项目
国家自然科学基金项目(32172178) （32172178）
