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
相关论文
共 30 条
  • [11] Overproduction of alkaline polygalacturonate lyase in recombinant Escherichia coli by a two-stage glycerol feeding approach
    Fang, Shuying
    Li, Jianghua
    Liu, Long
    Du, Guocheng
    Chen, Jian
    [J]. BIORESOURCE TECHNOLOGY, 2011, 102 (22) : 10671 - 10678
  • [12] Purification and properties of a thermoactive and thermostable pullulanase from Thermococcus hydrothermalis, a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent
    Gantelet, H
    Duchiron, F
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1998, 49 (06) : 770 - 777
  • [13] Localization of functional polypeptides in bacterial inclusion bodies
    Garcia-Fruitos, Elena
    Aris, Anna
    Villaverde, Antonio
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (01) : 289 - 294
  • [14] Highly thermostable amylase and pullulanase of the extreme thermophilic eubacterium Rhodothermus marinus:: production and partial characterization
    Gomes, I
    Gomes, J
    Steiner, W
    [J]. BIORESOURCE TECHNOLOGY, 2003, 90 (02) : 207 - 214
  • [15] Hii Siew Ling, 2012, Enzyme Res, V2012, P921362
  • [16] Inhibition of protein aggregation in vitro and in vivo by a natural osmoprotectant
    Ignatova, Zoya
    Gierasch, Lila M.
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (36) : 13357 - 13361
  • [17] An amyloid-forming segment of β2-microglobulin suggests a molecular model for the fibril
    Ivanova, MI
    Sawaya, MR
    Gingery, M
    Attinger, A
    Eisenberg, D
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (29) : 10584 - 10589
  • [18] Production of soluble recombinant proteins in Escherichia coli: Effects of process conditions and chaperone co-expression on cell growth and production of xylanase
    Jhamb, Kamna
    Sahoo, Debendra K.
    [J]. BIORESOURCE TECHNOLOGY, 2012, 123 : 135 - 143
  • [19] Molecular cloning and biochemical characterization of a heat-stable type I pullulanase from Thermotoga neapolitana
    Kang, Jinho
    Park, Kyung-Min
    Choi, Kyoung-Hwa
    Park, Cheon-Seok
    Kim, Go-Eun
    Kim, Doman
    Cha, Jaeho
    [J]. ENZYME AND MICROBIAL TECHNOLOGY, 2011, 48 (03) : 260 - 266
  • [20] An Approach to the Production of Soluble Protein from a Fungal Gene Encoding an Aggregation-Prone Xylanase in Escherichia coli
    Le, Yilin
    Peng, Jingjing
    Wu, Huawei
    Sun, Jianzhong
    Shao, Weilan
    [J]. PLOS ONE, 2011, 6 (04):
123