fulltext.study @t Gmail

High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
35010 45067 2010 5 PDF Available
Title
High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli
Abstract

Human insulin-like growth factor 1 (hIGF-1) is one kind of growth factor with clinical significance in medicine. The expression of TrxA-hIGF-1 fusion protein was rationally compared in three different Escherichia coli hosts (BL21 (DE3), Rosetta (DE3) and Rosetta-gami (DE3)) with the transformation of plasmid pET32-hIGF-1. The highest productivity of soluble hIGF-1 fusion protein was achieved in E. coli Rosetta-gami (DE3). Moreover, the effects of different expression conditions in this E. coli Rosetta-gami (DE3)/pET32-hIGF-1 host were systematically investigated to improve the expression level of the fusion protein. Under the optimized conditions, a high percent of the target fusion protein (96%) was expressed as soluble form with the volumetric production of soluble fusion protein reaching up to 2.06 g/L. After cell disruption, the soluble fusion protein was separated effectively by affinity chromatography and cleaved by enterokinase, with the concentration of mature hIGF-1 reaching up to 0.42 g/L in the mixture. The present work should be useful for the enhanced production of soluble protein with multiple disulfide bonds in E. coli.

Keywords
Insulin-like growth factor; hIGF-1; Soluble fusion expression; E. coli Rosetta-gami (DE3); TrxA
First Page Preview
High-level soluble expression of hIGF-1 fusion protein in recombinant Escherichia coli
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 45, Issue 8, August 2010, Pages 1401–1405
Authors
, , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering