High-level expression and purification of active human FGF-2 in escherichia coli by codon and culture condition optimization

Abstract EN

Basic fibroblast growth factor (bFGF) is a member of a highly conserved superfamily of proteins that are involved in cell proliferation, differentiation, and migration.

Objectives: The objective of this study was to overexpress and purify the high-level active human bFGF in Escherichia coli (E.

coli).

Materials and Methods: This experimental study was conducted in the Islamic Republic of Iran.

After codon optimization and gene synthesis, the optimized FGF-2 gene was subcloned into plasmid pET-32a.

pET32-FGF-2 was transformed into E.

coli BL21 for expression.

The cultivation parameters were optimized to produce a high yield of FGF-2.

Results: The optimal conditions were determined as follows: cultivation at 37°C in TB medium, with 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG), followed by post-induction expression for 6 h.

Under the abovementioned conditions, the expression volumetric productivity of FGF-2 reached 1.48 g/L.

Conclusions: A fusion tag from the pET32 expression plasmid permits the recovery of the recombinant fusion FGF-2 from E.

coli, without affecting its biological activity.