血链素的提纯和初步鉴定  

Purification and Characterization of Sanguicin——a Bacteriocin Produced by Streptococcus Sanguis

作　　者：邓辉[1] 丁一[1] 傅明德[2] 肖晓蓉[3] 刘娟[1] 周婷[1] 

机构地区：[1]四川大学华西口腔医学院口腔内科,成都610041 [2]四川大学华西基础医学与法医学院生物化学与分子生物学教研室 [3]四川大学华西口腔医学院教育部口腔生物医学工程重点实验室

出　　处：《四川大学学报（医学版）》2004年第4期555-558,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金 (批准号 3 960 0 163 )资助

摘　　要：目的　提纯血链素 ,为进一步从分子水平研究血链球菌及血链素对牙周可疑致病菌的作用机理提供实验基础。方法　利用 DEAE- Sepharose梯度洗脱和葡聚糖凝胶过滤相结合的方法 ,将 4 72 g/L 硫酸铵沉淀的血链素粗提物进一步提纯 ,测定其抑菌活性 ,并进行初步鉴定。结果　提纯获得了纯化的血链素 ,初步鉴定显示其具有蛋白质的性质 ,SDS- PAGE电泳结果显示主要区带一条 ,相对分子质量为 6 5× 1 0 3。体外抑菌试验显示提纯物对牙龈卟啉单胞菌、中间普雷沃菌和具核梭杆菌具有较强抑菌能力。结论　纯化血链素具有拮抗牙周可疑致病菌的重要作用 。Objective Streptoccus sanguis plays an important Role in maintaining the periodontal microecological balance. Previous studies have demonstrated that sanguicin——a kind of bacteriocin produced by S. sanguis has a prominent function of inhibiting the growth of putative periodontopathic bacteria (PPB). The aim of this study was to purify sanguicin and investigate its characters. Methods The raw extract of sanguicin was obtained from cells of S.sanguis by ultrasonication, salting out and dialysis. Then the diethyl aminoethyl sepharose gradual washing was done in conjunction with glucosan gel filtration for in the purfication of sanguicin. The characters of purified sanguicin were determined and its inhibitiory effect on PPB was measured by agar diffusion test. Results The purifed sanguicin was obtained; it was heat labile and proteinaceous; and by SDS PAGE, it showed a main band with the relative molecular mass of 65×10 3. The purifed sanguicin had a strong inhibitory effect on P.gingivalis, P.intermedia and F.nucleatum in vitro. Conclusion Our methods are useful for the purification of sanguicin. Sanguicin has inhibitory effect on PPB obviously and will have a good prospect in periodontal ecological therapy.

关 键 词：血链球菌 血链素 纯化SDS-聚丙烯酰胺凝胶电泳 

分 类 号：R781.4[医药卫生—口腔医学]