1.College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China; 2.State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, Xiamen University, Xiamen 361102, China
Abstract: A hybrid antimicrobial peptide sequence ofscygonadin-hepcidin3(scy-hep3) was obtained by overlap extension PCR using cDNA sequences of Scygonadin(Scy) and Hepcidin 3(Hep3) as the templates, and cloned into yeastPichiapastorisby the secreted expression vector of pPIC9K. The constructed plasmid of pPIC9K-scy-hep3 was transformed into the yeast GS115 cells by electroporation, and the hybrid antimicrobial peptide Scy-Hep3 was obtained and its antibacterial activity was analyzed after methanol induction and affinity purification in order to obtain stable expression of a new antibacterial peptide with high antibacterial activity. The results showed that the recombinant plasmid pPIC9K-scy-hep3 was constructed successfully, and the clone of GS115/pPIC9K-scy-hep3 was induced by 0.5% methanol for 24 h, a stable expressed product with molecular weight of about 24 000 being obtained, consistent with the calculated molecular weight of Scy-Hep3. Antibacterial assays demonstrated that the recombinant product of Scy-Hep3 had broad spectra of antimicrobial properties, especilally antibacterial activity against three Gram-positive bacteriaMicrococcuslysodeikticus,CorynebacteriumglutamicumandStaphylococcusaureus, and four Gram-negative bacteriaPseudomonasstutzeri,Pseudomonasfluorescens,AeromonashydrophilaandShigellaflexneri[minimal inhibitory concentration(MIC)<12 μmol/L]. There was higher antibacterial activity in the hybrid antibacterial peptide Scy-Hep3 than that in the single peptide of Scy and Hep3. The findings are first report on the hybrid antimicrobial peptide Scy-Hep3, and beneficial to production and application of Scy-Hep3 in future.
2018, Vol. 33 
