兽疫链球菌变异株产生的透明质酸的纯化及表征  

Purification and Structure Characterization of the Hyaluronic Acid Produced by a Streptococcus Zooepidemicus Mutation Strain

作　　者：洪水声[1] 陈佳[1] 滕利荣[1] 孟庆繁[1] 张建国[2] 刘兰英[1] 

机构地区：[1]吉林大学生命科学学院,长春130023 [2]中国科学院长春应用化学研究所电分析化学国家重点实验室,长春130022

出　　处：《高等学校化学学报》2004年第5期853-857,共5页Chemical Journal of Chinese Universities

基　　金：国家高等教育科学"十五"规划项目 (批准号 :2 0 0 2 ;2 0 -5 8-4 7);吉林省教育科学十五规划重点项目(批准号 :2 0 0 1-11-0 15 3 3 8)资助

摘　　要：用 N-甲基 -N′-硝基 -N-亚硝基胍 ( NTG)对兽疫链球菌进行诱变 ,获得高产菌株 .经过对该菌株的发酵培养 ,将产生的多糖经 Savage法、季铵复合物沉淀法、DEAE-纤维素 ( DE5 2 )离子交换层析法及 SephadexG-75凝胶过滤法分离纯化 .纯化的多糖结构经化学组成分析、核磁共振波谱、红外光谱及圆二色谱鉴定 ,证明了诱变得到的高产菌株 ( Streptococcuszooepidemicus J1 8)再经发酵 ,得到的多糖为透明质酸 .通过刚果红实验证明了透明质酸的构象为单股螺旋结构 ,其平均分子量约为 1 .1 6× 1 0 6 .A mutated strain of Streptococcus zooepidemicus capable of producing the polysaccharide in a high yield was obtained after being mutated by the chemical agent N-methyl-N′-nitro-N- nitrosoguanidine(NTG). Polysaccharide produced by the mutated strain of Streptococcus zooepidemicus was purified by the procedure including Savage method, quaternary ammonium compound precipitation, DEAE-cellulose(DE52) column chromatograph and Sephadex G-75 gel filtration. The purified polysaccharide was proved to be hyaluronic acid(HA) by means of chemical composition analysis, 13C NMR, Infrared spectra, and circular dichroism. The single helix conformation of the purified HA was studied by Congo red experiment. The molecular weight of the HA was about 1.16×10 6, which was measured by viscosity method.

关 键 词：病原菌 链球菌 变异株 透明质酸 分离 纯化 表征 诱变育种 发酵 

分 类 号：TQ929[轻工技术与工程—发酵工程]