高密度发酵重组大肠杆菌产海藻糖合成酶  

High cell density cultivation of Escherichia coli for the production of trehalose synthase

作　　者：晏星辰 黄璞 王鑫沂 姜艳芝 李斯斯 辛松 江凌[2] 黄和[3] YAN Xingchen;HUANG Pu;WANG Xinyi;JIANG Yanzhi;LI Sisi;XIN Song;JIANG Ling;HUANG He(College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211800,China;College of Food Science and Light Industry,Nanjing Tech University,Nanjing 211800,China;School of Pharmaceutical Sciences,Nanjing Tech University,Nanjing 211800,China)

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211800 [2]南京工业大学食品与轻工学院,江苏南京211800 [3]南京工业大学药学院,江苏南京211800

出　　处：《生物加工过程》2019年第4期385-391,共7页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金联合基金(U1603112);江苏省环保厅项目(2016053);江苏省“六大人才高峰”高层次人才项目(2015-JY-009);江苏省自然基金面上基金(BK20171461);江苏省先进生物制造创新中心项目(XTE1838)

摘　　要：酶法催化是目前生产海藻糖的主要手段。本文中,笔者通过高密度发酵重组大肠杆菌产海藻糖合成酶,进而以麦芽糖为底物,催化生产海藻糖。首先根据大肠杆菌高密度发酵条件要求,在揺瓶中对基因工程菌进行了培养基、发酵条件和诱导条件的逐一优化;然后在5L和50L发酵罐中进行批次补料发酵的放大实验;最后采用变指数恒pH法的策略发酵重组大肠杆菌,结果OD600达到97,海藻糖合成酶酶活达到(24000±350)U/mL,实现了海藻糖合成酶的重组大肠杆菌高密度发酵生产,极大提高了海藻糖的生产效率。Enzymatic catalysis is the main process for the trehalose production.In this study,we used high cell density cultivation of recombinant Escherichia coli to produce trehalose synthase and synthesized trehalose using maltose as a substrate.Firstly,according to conditions of high cell density culture of E.coli,we optimized the culture medium,fermentation conditions and induction conditions for producing trehalose synthase.Then,the scale-up experiments of fed-batch fermentation were conducted in 5 L and 50 L fermentors.Finally the fed-batch fermentation strategy-changed exponential-pH-stat feeding was carried out to produce trehalose synthase.The cell density and enzyme activity was 97 (OD600) and (24 000±350) U/mL,respectively,which greatly improved the production efficiency of trehalose.

关 键 词：海藻糖 海藻糖合成酶 大肠杆菌 高密度发酵 变指数恒pH法 

分 类 号：Q939.9[生物学—微生物学]