Optimization of expression conditions for signal molecule AI-2 synthetic protein LuxS by response surface methodology
Quorum sensing is a density-dependent gene expression regulation method mediated by autoinducers(AIs)in bacteria,which has been proven to participate in the regulation of multiple physiological functions of bacteria.The LuxS/AI-2 type quorum sensing system mediated by signal molecule AI-2 is widely present in Gram positive bacteria and negative bacteria,which has received considerable attentions.The synthesis of AI-2 depends on the catalytic action of S-ribosylhomocysteinase(LuxS),and the LuxS protein is obtained through transcription and translation of the luxS gene.The purpose of this study is to explore the optimal expression conditions of LuxS protein in the expression strain Escherichia coli BL21(DE3),thereby ob-taining a high yield of LuxS protein with certain biological activity.This study first predicted the structure of LuxS protein and determined that E.coli BL21(DE3)was used as the expression strain.Secondly,response surface methodology was further used to optimize the expression conditions of LuxS protein based on single factor optimization.The highest expression condition of LuxS protein was obtained under the conditions of bacterial concentration with OD600 of 0.5,induction temperature of 37 ℃,IPTG concentration of 0.5 mmol/L,and induction time of 31 h.This study successfully optimized the expression conditions of LuxS protein in E.coli BL21(DE3),and obtained high yield of LuxS protein with biological activity,laying a foundation for the synthesis of AI-2 in vitro.
quorum sensingAI-2LuxS proteinprokaryotic expressionoptimization of conditions
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维普
