κ-卡拉胶寡糖的酶解制备及其体外抗病毒活性

Preparation and in Vitro Antiviral Activity of kappa-Carrageenan Oligosaccharides

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.