Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2078994 | Chinese Journal of Biotechnology | 2007 | 5 Pages |
Abstract
Hepcidin is a liver-expressed, small peptide rich in cysteine that acts as a regulator of systemic iron homeostasis. In this work, according to the partiality codon of Pichia pastoris, a DNA fragment containing the coding sequence of hepcidin was designed and synthesized, especially a Kex2 signal cleavage site was fused in the 5' end of the antibacterial peptide genes. The modified hepcidin gene was then inserted into the P. pastoris expression vector plasmid pPICZα-A. After electroporation of the resulting vector, pPICZα-A-Hepc, into the yeast host strain GS115, transformants with high copy inserts were selected by 1500 mg/L Zeocin⢠selection. Under the control of the promoterAOX1 (alcohol oxidase 1), recombinant hepcidin secreted from P. pastoris had a molecular weight of 2.7 kD. After optimization of the flask-shaking culture fermentation, the yield of hepcidin reached 100 mg/L in the clarified broth. Through antibacterial assay, the recombinant hepcidin displayed obvious antibacterial activity against Bacillus subtilis. But it could not distinctly inhibit the growth of E. coliBL21(DE3).
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Authors
Hui ZHANG, Qi-Peng YUAN,