Optimization of pullulanase production in Escherichia coli by regulation of process conditions and supplement with natural osmolytes

被引：54
作者：
Duan, Xuguo [1 ,2 ,3 ]
Chen, Jian [1 ,2 ,3 ]
Wu, Jing [1 ,2 ,3 ]
机构：
[1] Jiangnan Univ, State Key Lab Food Sci & Technol, 1800 Lihu Ave, Wuxi 214122, Peoples R China
[2] Jiangnan Univ, Minist Educ, Sch Biotechnol, Wuxi 214122, Peoples R China
[3] Jiangnan Univ, Minist Educ, Key Lab Ind Biotechnol, Wuxi 214122, Peoples R China
来源：
BIORESOURCE TECHNOLOGY | 2013年 / 146卷
基金：
中国国家自然科学基金;
关键词：
Escherichia coli; Pullulanase; Soluble expression; Betaine; I-PULLULANASE; THERMOSTABLE PULLULANASE; THERMOLEOVORANS US105; RECOMBINANT PROTEINS; MOLECULAR CHAPERONES; EXPRESSION; CLONING; STARCH; VITRO; GENE;
D O I：
10.1016/j.biortech.2013.07.074
中图分类号：
S2 [农业工程];
学科分类号：
0828 ;
摘要：
In this study, the effects of temperature, IPTG (Isopropyl beta-D-1-thiogalactopyranoside) concentration, and osmolytes (proline, K-glutamate, and betaine) on cell growth and soluble pullulanase productivity of recombinant Escherichia coli were investigated. The yield of soluble pullulanase was found to be enhanced with decrease in cultivation temperature, lower IPTG concentration, and betaine supplementation in a shake flask. In addition, a modified two-stage feeding strategy was proposed and applied in a 3-L fermentor supplied with 20 mM betaine, which achieved a dry cell weight of 59.3 g L-1. Through this cultivation approach at 25 degrees C, the total soluble activity of pullulanase reached 963.9 U mL(-1), which was 8.3-fold higher than that observed without addition of betaine at 30 degrees C (115.8 U mL(-1)). The higher expression of soluble pullulanase in a scalable semisynthetic medium showed the potential of the proposed process for the industrial production of soluble enzyme. (c) 2013 Elsevier Ltd. All rights reserved.
引用
收藏
页码：379 / 385
页数：7
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