High-level Production of Creatine Amidinohydrolase from Arthrobacter nicotianae 23710 in Escherichia coli

被引:5
作者
Dai, Jun [1 ,2 ]
Zhang, Linpei [1 ,2 ]
Kang, Zhen [1 ,2 ]
Chen, Jian [2 ,3 ]
Du, Guocheng [2 ,4 ]
机构
[1] Jiangnan Univ, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Peoples R China
[2] Jiangnan Univ, Sch Biotechnol, Wuxi 214122, Peoples R China
[3] Jiangnan Univ, Natl Engn Lab Cereal Fermentat Technol, Wuxi 214122, Peoples R China
[4] Jiangnan Univ, Key Lab Carbohydrate Chem & Biotechnol, Minist Educ, Wuxi 214122, Peoples R China
基金
美国国家科学基金会; 国家高技术研究发展计划(863计划);
关键词
Creatine amidinohydrolase; Arthrobacter nicotianae; Escherichia coli; Two-stage control strategy; PSEUDOMONAS-PUTIDA; RECOMBINANT PROTEIN; DIRECTED EVOLUTION; SARCOSINE OXIDASE; EXPRESSION; CLONING; GENE; PURIFICATION; CRYSTALLIZATION; STRATEGIES;
D O I
10.1007/s12010-014-1460-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the present study, the gene encoding creatinase was amplified from Arthrobacter nicotianae 23710 (CICC) and functionally overexpressed in Escherichia coli. By applying a two-stage temperature control strategy, the production of creatinase was increased up to 61.3 U/mL in 3-L fermentor with a high productivity of 6.1 U/mL/h. The recombinant creatinase shows excellent resistance to the chelating agent EDTA, the surfactants (Tween 20, Tween 80, and Triton X-100) and the common preservative NaN3 (20 mM). High-level expression of the recombinant creatinase will contribute to its application in clinical diagnosis of renal function.
引用
收藏
页码:2564 / 2573
页数:10
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