解淀粉芽孢杆菌Q-426群体感应系统ComX信息素前体肽的合成研究  

Synthesis of the ComX Pheromone Precursor Peptide in Bacillus amyloliquefaciens Q-426 Quorum Sensing System

作　　者：吴懿[1] 张丽影[1] 权春善[1] 钟美玲[1] 王路路[2] 王冠天[1] 

机构地区：[1]大连民族大学生命科学学院生物技术与资源利用国家民委教育部重点实验室,大连116600 [2]大连理工大学化工与环境生命学部,大连116024

出　　处：《中国生物工程杂志》2017年第5期38-44,共7页China Biotechnology

基　　金：国家自然科学基金(21272031;21372037);中央高校自主科研基金(DC201502020201;DC201501078)资助项目

摘　　要：采用Fmoc固相合成法合成了解淀粉芽孢杆菌Q-426群体感应系统Com X信息素的前体五肽。利用反向半制备色谱(RP-HPLC)和液质联用仪(LC-MS)对合成的五肽进行了分离纯化及纯度分析。建立了检测游离氨基酸的新方法,并详细考察了合成条件对树脂上肽链连接效率的影响。结果表明,新型NNA试剂(茚三酮+正丁醇+乙酸溶液)可替代传统的Kaiser试剂用于Fmoc固相合成中游离氨基的检测;在35℃下,使用DCM和DMF对树脂交替溶胀1h后,树脂上肽链的连接效率最高。合成的Com X信息素前体五肽粗品产率为98%,纯度为58.60%;经半制备型高效液相纯化后,纯度达99%。抑菌实验表明,Com X信息素前体五肽具有明显的生物活性。The ComX pheromone precursor peptide of Bacillus amyloliquefaciens Q-426 quorum sensing system was successfully synthesized by Fmoc solid-phase synthesis method, and then isolated and purified by semi preparative chromatography （RP-HPLC） and liquid chromatography （HPLC-MS）. A new method for the determination of free amino groups was established, and the effects of swelling process and reaction temperature on the degree of substitution were also investigated. The results showed that the traditional Kaiser reagent for the detection of free amino Fmoc in solid-phase synthesis can be replaced by the NNA reagent （ ninhydrin ＋ N- butanol ＋ acetic acid solution）. And the resins have the highest substitution degree after they were alternate swelling by DCM and DMF for lh at 35℃. The product yield and the purity of the crude precursor peptide of ComX pheromone is 98% and 58.60% respectively. And the purity of the precursor peptide of ComX pheromone is 99% after being purified by semi preparative high performance liquid. Antibacterial experiments showed that the precursor peptide of ComX pheromone had obvious biological activity.

关 键 词：Fmoc固相合成 ComX信息素前体五肽 氨基检测 分离纯化 

分 类 号：Q816[生物学—生物工程]