卷曲乳杆菌类细菌素的分离纯化及表征  

Isolation,purification and characterization of bacteriocin-like substance from Lactobacillus crispatus

作　　者：邹达 唐美 黄雄辉 邹俊娜 王艳[1] ZOU Da;TANG Mei;HUANG Xiong-hui;ZOU Jun-na;WANG Yan(Chemistry and Chemical Engineering of Central South University,Changsha 410083)

机构地区：[1]中南大学化学化工学院,长沙410083

出　　处：《中南药学》2023年第7期1744-1750,共7页Central South Pharmacy

摘　　要：目的初步分离纯化并鉴定卷曲乳杆菌类细菌素,为研发针对念珠菌性阴道炎(VVC)的药物奠定基础。方法以氟康唑作为标准药物,测定卷曲乳杆菌发酵上清液(CFS)对于白假丝酵母菌的抑菌活性,研究温度、pH值和酶解处理对于CFS抑菌活性的影响;采用硫酸铵沉淀法、疏水柱层析法及透析法对CFS中的抑菌活性物质进行初步分离纯化;采用苯酚-硫酸法、双缩脲反应、UV、FTIR、HPLC和SDS-PAGE对抑菌活性物质进行初步鉴定。结果卷曲乳杆菌CFS中存在耐酸碱、热稳定性好的糖蛋白或糖肽类抑菌物质,初步纯化之后CFS比活力从218.6 U·mg^(-1)提升到700.7 U·mg^(-1),提高了3.2倍,初步鉴定为分子量均为55 kD左右的两种糖蛋白或糖肽。结论卷曲乳杆菌CFS中含有糖蛋白或糖肽类细菌素,对白假丝酵母菌具有抑制作用,有望将其开发成药物。Objective To isolate and identify bacteriocin-like substances produced by the Lactobacillus crispatus,and lay the foundation for research and drug development for vulvovaginal candidiasis(VVC).Methods Fluconazole was used as the standard drug,the bacteriostatic activity of cell-free supernatant of Lactobacillus crispatus(CFS)for Candida albicans was determined.The effect of temperature,pH value and enzymatic hydrolysis on the antibacterial activity of CFS was determined.The antibacterial active substances in CFS were preliminarily separated and purified by ammonium sulfate precipitation,hydrophobic column chromatography and dialysis.Phenol-sulfuric acid method,biuret test,UV,FTIR,HPLC and SDS-PAGE were used to identify the antibacterial active substances.Results Glycoprotein or glycopeptide antibacterial substances were found with good acid-base resistance and thermal stability in CFS of Lactobacillus crispatus.After the purification,the specific activity increased from 218.6 U·mg^(-1) to 700.7 U·mg^(-1),by 3.2 times.Two glycoproteins with molecular weight of about 55 kD were identified.Conclusion CFS of Lactobacillus crispatus contains glycoprotein or glycopeptide bacteriocin,which can inhibit Candida albicans and as a prospective drug.

关 键 词：卷曲乳杆菌 类细菌素 分离纯化 表征 

分 类 号：R915[医药卫生—微生物与生化药学]