摘要
利用Pseudoalterom onassp.AJ5-13菌株所产生的κ-卡拉胶酶降解κ-卡拉胶制备κ-卡拉胶寡糖,通过电喷雾离子化飞行时间质谱(ESI-TOF-MS)和核磁共振波谱(13C-NMR)分析,该酶的水解产物主要是硫酸κ-新卡拉二糖、硫酸κ-新卡拉四糖、硫酸κ-新卡拉六糖、硫酸κ-新卡拉八糖和硫酸κ-新卡拉十糖,确定该κ-卡拉胶酶专门水解κ-卡拉胶3,6-内醚-D-半乳糖和4-硫酸-D-半乳糖之间的β-1,4糖苷键,产生3,6-内醚-D-半乳糖作为非还原端,D-半乳糖作为还原端的κ-新卡拉寡糖。采用体外Vero细胞培养法和四甲基偶氮唑盐(MTT)比色法研究了寡糖抗标准单纯疱疹病毒1型(HSV-1)活性。结果表明,κ-新卡拉寡糖对Vero细胞毒性极低,可干扰HSV-1毒株向Vero细胞的吸附。
κ-Carrageenan oligosaceharides were prepared by the method of enzymatic hydrolysis using κ- carrageenase from Pseudoalteromonas sp. AJ5 - β. With the analysis of composition and structure by u- sing ESI-TOF-MS and 13C-NMR, the main hydrolyzed products were demonstrated as κ-neocarrabiose, - tetraose, -hexaose, -oetaose, and -decaose sulfates with 3-linked 13-D-galactopyranose 4-sulfate as the reducing end. It indicated that the κ-carrageenase from Pseudoalteromonas sp. AJ5-z13 specifically hydrolyzed the 13-1, 4 glycosidic linkages of κ-carrageenan, producing κ-neocarraoligosaccharides with the 3, 6- anhydro-D-galaetose as the non reducing end and D-galactose as the reducing end. The anti-HSV-1 activities of κ-neocarraoligosaccharides in vitro were determined by Vero cell culture and MTT assay. The results showed that κ-neocarraoligosaccharides had little cytotoxic effect on Vero ceils and could interfere the absorption of HSV-1 to Vero cells.
出处
《中山大学学报(自然科学版)》
CAS
CSCD
北大核心
2009年第6期105-108,137,共5页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
国家自然科学基金资助项目(40506027)
农业部海洋水产增养殖学与生物技术重点开放实验室开放课题资助项目(K2006-12)