响应面法优化信号分子 AI-2合成蛋白 LuxS表达条件
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摘要：
群体感应是一种由自诱导物(autoinducers,A1s)介导的、细菌密度依赖的基因表达调控方式,已经被证实参与细菌多种生理功能的调控。信号分子A1-2介导的LuxS/A1-2型群体感应系统广泛存在于革兰氏阳性和阴性细菌,并备受关注,AL2 的合成依赖于 S-核糖同型半胱氨酸酶(LuxS)的催化作用,而 LuxS蛋白则是由 luxS 基因经过转录、翻译得到的,为了探索 LuxS 蛋白在表达菌株 Escherichia coli B21(DE3)中的最佳表达条件,从而获得较高产量且具有一定的生物活性的 LuxS 蛋白,首先对 LuxS蛋白结构进行预测,确定 E.coi BL21(DE3)作为表达萌株,其次,在单因素优化的基础上,进一步通过响应面法优化 Luxs 蛋白的表达条件,在菌液浓度 ODw为 0.5,诱导温变为 37 ℃、IPTG 浓度为 0.5 mmol/1,,诱导时间为31h条件下,获得Luxs 蛋白的最高表达量,研究成功优化了Luxs蛋白在 E.coli BL21(DE3)中的诱导表达条件,获得了具有生物活性的LuxS蛋白,并在后续的研究中成功合成了具有生物活性的信号分子 A1-2,为体外合成 AI-2 奠定了基础.
Quorum sensing is a density-dependent gene expression regulation method mediated by autoinducers (Als) in bacteria, which has been proven to participate in the regulation of multiple physiological functions of bacteria. The luxS/AF2 type quorum sensing system meliated by signal molecule Al-2 is widely present in Gram positive bacteria and negative bacteria which has received considerable attentions, 'The synthesis of AF2 depends on the catalytic action of $-ribosylhomocysteinase (LuxS), and the LuxS protein is obtained through transeription and translation of the luxS gene. The purpose of this study is to explore the optimal expression conditions of LuxS protein in the expression strin Escherich ia coi B2l(DE3), thereby ob.taining a high yield of luxS protein with certain biologieal activity. This study first predicted the structure of LuxS protein and determined that E. coli Bl.2l(DE3) was used as the expression strain, Secondly, response surlace methodology was further used to optimize the expression conditions of LuxS protein based on single factor optimization. The highest expression condition of Lux$ protein was obtained under the conditions of bacterial concentration with ODw, of 0.5, induction temperature of 37 IP'TG concentration of 0.5 mmol/L, and induetion time of 31 h, This study suecesfully optimized the expression conditions of LuxS protein in E. coli Bl21(DE3), and obtained high yield of Lux$ protein with biological activity, laying a foundation for the synthesis of Al-2 in vitro.
作者：
李祎,韩朔,王玉琪,秦梦园,吴晓敏
LiYi,HanShuo,WangYuqi,QinMengyuan,WuXiaomin
机构地区：
Betway官方客服生命科学学院
引用本文：
李祎,韩朔,王玉琪等.响应面法优化信号分子 AI-2合成蛋白 LuxS表达条件[J].Betway官方客服学报(自然科学版),2025,53(1):136-144.(LiYi,HanShuo,WangYuqi,etal.Optimizationofexpressionconditionsforsignal
moleculeAI-2syntheticproteinLuxSbyresponsesurfacemethodology[J].JournalofHenanNormalUniversity
(NaturalScienceEdition),2025,53(1):136-144.DOI:10.16366/j.cnki.1000-2367.2023.03.29.0001.)
基金：
国家自然科学基金
关键词：
群体感应;AI-2;LuxS蛋白;原核表达;条件优化
quorum sensing: Al-2: LuxS protein: prokaryotic expression: optimization of conditions
分类号：
Q93
