High level expression of recombinant BoNT/A-Hc by high cell density cultivation of Escherichia coli

被引：22

作者：

Yari, Kheirollah ^{[1
,2
]}

Fatemi, Seyed Safa-Ali ^{[1
]}

Tavallaei, Mahmood ^{[3
]}

机构：

[1] NIGEB, Dept Ind & Environm Biotechnol, Tehran, Iran

[2] Kermanshah Univ Med Sci, Med Biol Res Ctr, Kermanshah, Iran

[3] Baqiyatallah Univ Med Sci, Genet Res Ctr, Tehran, Iran

来源：

BIOPROCESS AND BIOSYSTEMS ENGINEERING | 2012年 / 35卷 / 03期

关键词：

Clostridium botulinum; Escherichia coli; High cell density cultivation; Taguchi method; Fed-batch; BOTULINUM NEUROTOXIN SEROTYPE; HUMAN INTERFERON-GAMMA; FERMENTATION; PURIFICATION; SELECTION;

D O I：

10.1007/s00449-011-0579-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The carboxylic domain of the Clostridium botulinum neurotoxin heavy chain (BoNT/A-HC), which has been reported as a vaccine candidate, contains the principle protective antigenic determinants. In this study, the high level expression of the BoNT/A-Hc was achieved by high cell density cultivation of recombinant Escherichia coli in a 2-l batch stirred-tank bioreactor. In order to maximize protein expression, post-induction time and IPTG inducer concentration were optimized by the Taguchi statistical design method. Results showed that the middle of the logarithmic phase and an IPTG concentration of 1 mM presented the optimum conditions for the maximum expression of BoNT/A-HC. High cell density cultivation was subsequently carried out as an effective strategy for the high level expression of recombinant BoNT/A-Hc. Consequently, soluble BoNT/A-Hc was produced at the maximum level of 486 mg l(-1), at 3 h post-induction, which was approximately 9.3 and 7.8 times higher than the levels produced by the shake flask and batch culturing methods, respectively.

引用

页码：407 / 414

页数：8

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