Article ID Journal Published Year Pages File Type
35010 Process Biochemistry 2010 5 Pages PDF
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.

Related Topics
Physical Sciences and Engineering Chemical Engineering Bioengineering
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