Expression and purification of a fusion-typed pediocin PA-1 in Escherichia coli and recovery of biologically active pediocin PA-1

被引：26

作者：

Moon, GS

Pyun, YR

Kim, WJ ^{[1
]}

机构：

[1] Korea Food Res Inst, Food Safety Res Div, Gyeonggi 463746, South Korea

[2] Yonsei Univ, Dept Biotechnol, Seoul 120749, South Korea

来源：

INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY | 2006年 / 108卷 / 01期

关键词：

pediocin PA-1; overexpression; fusion protein; heterologous expression;

D O I：

10.1016/j.ijfoodmicro.2005.10.019

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

Pediocin PA-1 is a representative class IIa bacteriocin which is small and heat-stable and has a consensus motif, -YGNGV-. The plasmid pQE40PED, encoding pediocin PA-1 fused with His-tagged mouse dihydrofolate reductase (DHFR), was constructed and introduced into Escherichia coli M15 strain. The fusion protein was overexpressed in the strain after induction of isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography. For the recovery of biologically active pediocin PA-1, the purified fusion protein was cleaved by Factor Xa protease and the liberated pediocin PA-1 was finally purified by ultrafiltration with a 75% yield. The molecular mass of the purified recombinant pediocin PA-1 was the same as that of native pediocin PA-1 oil an electrophoresis gel. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：136 / 140

页数：5

共 24 条

[1]

Ausubel F.M., 1991, CURRENT PROTOCOLS MO

[2] ANALYSIS OF THE PEDIOCIN ACH GENE-CLUSTER FROM PLASMID PSMB74 AND ITS EXPRESSION IN A PEDIOCIN-NEGATIVE PEDIOCOCCUS-ACIDILACTICI STRAIN [J].