Heterologous expression of human KGF/FGF7 (Keratinocyte growth factor) in Pichia pastoris

Abstract

Keratinocyte Growth Factor (KGF) is a paracrine-acting and epithelium-specific growth factor produced by cells of mesenchymal origin, play an important role in promoting proliferation, differentiation, motility of epithelial cells and stimulating regeneration of damaged epithelial tissues. Recent studies indicated that recombinant KGF is produced in many different expression systems such as bacteria, insect cells, plant and mammalian cells. However, KGF’s yields obtained from these systems is low and production’s cost is high especially in mammalian cells. In this study, the yeast Pichia pastoris was chosen as a host for KGF expression through induction of methanol by promoter AOX on pPICzαA vector system. The results demonstrated that the Pichia pastoris X33:kgf transformants secreted KGF directly into BMMY medium after inducing by 0.5% methanol. The recombinant protein was purified by heparin affinity chromatography with the yield of 1.35 mg/l and the purity of 99.89% showed by SDS-PAGE. In addition, MTT assay showed the purified recombinant KGF had a proliferation effect on A549 cell line since A549 known as a cell has KGF’s receptor.