Cloning and expression of a bactericin sakacin P from Lactobacillus sakei in Escherichia coli

• 1、School of Food Science and Technology, Jiangnan University

Abstract：Sakacin P, a novel bactericin from Lactobacillus sakei, is a potential antimicrobial substrate, which has strong activity against food-borne pathogen eg. Listeria monocytogenes. In Lactobacillus sakei, the gene (sppA) of sakacin P was controled by strict regulation mechnism, so that the quantity of secreted sakacin P was low. In this study, we synthesized the sppA gene using splicing by overlap extension PCR (SOE-PCR), and cloned it into E. coli expression system. After the induction of IPTG, the recombinant sakacin P was successfully expressed. The collected cells were sonicated and the activity was identified by inhibition zone method using the cell-free supernatant. The results also showed that the low temperature introduction took more active sakacin P which has strong antimicrobial activity.

keywords： sakacin P expression class IIa bacteriocins

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