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Expression of antimicrobial peptide dybowskin-2CDYa and hEGF in pichia pastoris and characterization of its dual function

Li Qiang, Jin Lili, Wang Zheng, Yue Xiqing, Wang qiuyu

Dybowskin-2CDYa (Dy2) with broad spectrum antibacterial activity is a novel antimicrobial peptide identified fromskin of the frog Rana dybowskii. In order to get a dual function protein with inhibiting bacterial growth and promoting cell proliferation,we explored the fusion expression ofDy2 and human epidermal growth factor (hEGF) by yeast expression system. The fusion gene was constructed and cloned into the expression vector pPIC9K, transformed to competent P. pastorisGS115.The expressed fusion protein was purified from the culture supernatant by Sephadex G-50 and followed by C18 reverse phased HPLC and indentified by SDS-PAGE and antibacterial activity assay. Antimicrobial activity assay showed that the recombinant fusion protein could inhibit the growth of a broad spectrum of bacteria, while displaying a significant promoting effect on proliferation andmigration ofNIH3T3 cell lines compared with the control group. These results indicated that the expressed fusion protein was capable of killing baterial and promoting proliferation of cells, which suggest that it has a good prospect in the therapy of trauma and burns