Expression of porcine protegrin-1 in Pichia pastoris and its anticancer activity in vitro

• Published in: Experimental and therapeutic medicine 2015-03, Vol.9 (3), p.1075-1079
• Main Authors: NIU, MINGFU, CHAI, SHUMAO, YOU, XIAOYAN, WANG, WENHUI, QIN, CUILI, GONG, QIANG, ZHANG, TINGTING, WAN, PENG
• Format: Article
• Language: English
• Subjects: antibacterial peptide protegrin-1/His; anticancer activity; Antimicrobial peptides; Antimitotic agents; Antineoplastic agents; Bacteria; Bioengineering; Cancer; Cell culture; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA polymerase; E coli; Engineering; Enzymes; expression and purification; Gene expression; Lipids; Medical research; Medicine, Experimental; Peptides; Proteins; Studies; Yeast
• ISSN: 1792-0981; 1792-1015
• DOI: 10.3892/etm.2015.2202

Protegrin-1 (PG-1), a β-hairpin antimicrobial peptide (AMP), is amongst the shortest AMPs in sequence length while remaining active against a variety of microorganisms. The aim of this study was produce recombinant PG-1 and investigate its anticancer activity. A DNA sequence encoding the mature PG-1, fused with a 6His-tag, was cloned into the pPICZα-A vector and transformed into Pichia pastoris. Expression was induced following culture for ~96 h with 1% methanol at 28°C, and ~15.6 mg PG-1 was expressed in 100 ml culture medium. Following purification using a Ni-chelating Sepharose column, ~20 mg pure active PG-1 was obtained from 500 ml culture broth supernatant. The expressed PG-1/6His exhibited strong dose- and time-dependent anticancer activity against HepG2 cells in vitro.