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On-line Access: 2010-01-06

Received: 2010-04-26

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Crosschecked: 2010-12-09

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Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.1 P.65-71

http://doi.org/10.1631/jzus.B1000152


Fusion expression of pedA gene to obtain biologically active pediocin PA-1 in Escherichia coli


Author(s):  Shan-na Liu, Ye Han, Zhi-jiang Zhou

Affiliation(s):  School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

Corresponding email(s):   zzj@tju.edu.cn

Key Words:  Bacteriocin, Fusion expression, Inclusion body, Pediocin PA-1, Thioredoxin


Shan-na Liu, Ye Han, Zhi-jiang Zhou. Fusion expression of pedA gene to obtain biologically active pediocin PA-1 in Escherichia coli[J]. Journal of Zhejiang University Science B, 2011, 12(1): 65-71.

@article{title="Fusion expression of pedA gene to obtain biologically active pediocin PA-1 in Escherichia coli",
author="Shan-na Liu, Ye Han, Zhi-jiang Zhou",
journal="Journal of Zhejiang University Science B",
volume="12",
number="1",
pages="65-71",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000152"
}

%0 Journal Article
%T Fusion expression of pedA gene to obtain biologically active pediocin PA-1 in Escherichia coli
%A Shan-na Liu
%A Ye Han
%A Zhi-jiang Zhou
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 1
%P 65-71
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000152

TY - JOUR
T1 - Fusion expression of pedA gene to obtain biologically active pediocin PA-1 in Escherichia coli
A1 - Shan-na Liu
A1 - Ye Han
A1 - Zhi-jiang Zhou
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 1
SP - 65
EP - 71
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000152


Abstract: 
Two heterologous expression systems using thioredoxin (trxA) as a gene fusion part in Escherichia coli were developed to produce recombinant pediocin PA-1. pediocin PA-1 structural gene pedA was isolated from Pediococcus acidilactici PA003 by the method of polymerase chain reaction (PCR), then cloned into vector pET32a(+), and expressed as thioredoxin-PedA fusion protein in the host strain E. coli BL21 (DE3). The fusion protein was in the form of inclusion body and was refolded before purification by nickel-iminodiacetic acid (Ni-IDA) agarose resin column. Biological activity of recombinant pediocin PA-1 was analyzed after cleavage of the fusion protein by enterokinase. Agar diffusion test revealed that 512-arbitrary unit (AU) recombinant pediocin PA-1 was obtained from 1 ml culture medium of E. coli (pPA003PED1) using Listeria monocytogenes as the indicator strain. thioredoxin-PedA fusion gene was further cloned into pET20b(+). thioredoxin-PedA fusion protein was detected in both the periplasmic and cytoplasmic spaces. The recombinant pediocin PA-1 from the soluble fraction attained 384 AU from 1 ml culture medium of E. coli (pPA003PED2). Therefore, biologically active pediocin PA-1 could be obtained by these two hybrid gene expression methods.

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Reference

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