β-1,3-1,4葡聚糖酶在大肠杆菌中的高效分泌表达

Efficient secretory expression of β-1, 3-1, 4-glucanase in E.coli

摘要: 在大肠杆菌中实现β-1,3-1,4-葡聚糖酶的高效分泌表达。将实验室自主开发的信号肽ff53与β-1,3-1,4-葡聚糖酶成熟肽基因（bgl）进行融合连接到p ET-28a（+）上;通过优化诱导表达条件,28℃、8 g/L乳糖诱导10 h,重组菌E.coli BL21/p ET-ff53-bgl发酵液上清中β-1,3-1,4-葡聚糖酶活力达到1093 U/m L,与IPTG诱导的重组菌E.coli BL21/p ET-bgl（583 U/m L）相比,提高了0.87倍。本研究为高密度发酵制备该酶奠定了基础。

Abstract: In this study, the highly secretory expression of β- 1, 3- 1, 4- glucanase was achieved in E. coli. Signal peptide ff53 developed by our team was fused with mature peptide gene ( bgl) of β- 1, 3- 1, 4- glucanase, then cloned into the p ET- 28a ( +) plasmid. Efficient extracellular expression was obtained under following optimized conditions: 8g / L lactose at 28 ℃ for 10 h, resulting in the enzyme activity of E.coli BL21 / p ET- ff53- bgl in culture medium reached 1093 U / m L, which increased 0.87 folds compared with enzyme activity of E.coli BL21 / p ET- bgl ( 583 U/m L) induced with IPTG.The study laid the foundation of high density fermentation of β-1, 3-1, 4-glucanase production.