Synthesis of recombinant gene for Bacillus subtilis bacteriocin subtilosin A and its expression in Pichia pastoris

In this study,the recombinant gene for subtilosin A(rsbo A) was synthesis for the sake of the expression for this bacteriocin in host cell P. pastoris. Both the results of colony PCR of positive Escherichia coli recombinants and DNA sequencing indicated that recombinant plasmid p PIC9K- sbo was constructed successfully. In addition,the results of colony PCR of positive P. pastoris recombinants indicated that recombinant Pichia GS115- p PIC9K- sbo was constructed successfully. Protein electrophoresis analysis of positive P. pastoris recombinants displayed that the recombinant gene rsbo A was secretory expressed in P. pastoris GS115.Analysis of antimicrobial activity showed that the fermentation broth of this recombinant yeast possess antibacterial activities against P. aeruginosa. The result of this work built a new strategy of bacteriocins,a small molecule peptide,in heterologous expression. The study laid a foundation for industrialized production of Bacillus subtilis antibiotic peptide subtilosin A.