The 46th Academic Congress of Taiwan Association for Food Science and Technology

Expression of porcine antibacterial peptide PMAP-37 in Escherichia coli using SUMO fusion technology

YS Chou, HJ Lin, WC Huang, ZW Chen, JH Lin, SL Hsuan, JP Wang

PMAP-37 is an antibacterial peptide that could be isolated from porcine myeloid. The peptide is an amphiphilic α-helix with 37 amino acid residues. Previous study showed that PMAP-37 strongly inhibits the growth of several strains of Gram-negative and Gram-positive bacteria such as Escherichia coli, Pseudomonas aeruginosa, Bacillus megaterium and Staphylococcus aureus. The main objective of this study is to establish a method for production of recombinant PMAP-37 (rPMAP-37) in E. coli. The DNA sequence encoding the PMAP-37 was designed according to E.coli preferred codons and synthesized through overlapping extension polymerase chain reaction. The codon-optimized PMAP-37 gene was inserted into pET-SUMO which contains the small ubiquitin-like modifier (SUMO) gene for the construction of fusion expression vector. Then, the expression vector was transformed into E.coli. Our results showed that rSUMO-PMAP37 fusion protein was expressed in soluble form in E.coli and could be purified by immobilized metal ion affinity chromatography. This fusion protein could be digested by SUMO protease. The antibacterial activity of rPMAP37 will be further determined.