平颏海蛇皮中新型硫酸皮肤素的分离纯化和结构表征

目的 从平颏海蛇皮中提取分离糖胺聚糖(GAGs),并对其结构进行初步表征。方法 对海蛇皮进行脱脂处理后,采用木瓜蛋白酶和胰蛋白酶联合酶解提取粗多糖,采用氯代十六烷基吡啶(CPC)沉淀和Q-Sepharose Fast Flow离子交换树脂对粗多糖进行分离纯化。运用醋酸纤维素薄膜电泳分析其GAGs种类,采用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生高效液相色谱(HPLC)法分析其单糖组成,利用硫酸软骨素酶降解结合质谱和强阴离子交换色谱(SAX-HPLC)法分析其2糖组成。结果 获得了2种GAGs纯化组分,并对其中1种含量较高的纯化组分(HSAP-2)进行了初步结构表征。HSAP-2中含有4种2糖,主要为单硫酸化2糖(4S和6S)和少量2硫酸化2糖(46S)、非硫酸化2糖(0S)；单糖组成中含有葡萄糖醛酸(GlcA)和乙酰氨基半乳糖(GalNAc)；是1种高度杂合的新型硫酸皮肤素(DS)。结论 从平颏海蛇皮中分离纯化获得了1种结构新颖的DS,为平颏海蛇多糖结构与生物活性的深入研究提供了基础。

Isolation, purification and structural characterization of a novel dermatan sulfate from Lapemis curtus skin

Objective To extract and isolate glycosaminoglycans (GAGs) from Lapemis curtus skin, and to preliminarily investigate its structural features. Methods The skin of Lapemis curtus was defatted and digested by papain-trypsase to release glycans. Then the crude polysaccharide was separated by cetylpyridinium chloride (CPC) deposition and Q Sepharose Fast Flow anion-exchange chromatography. The GAGs types of HSAP-2 were determined by cellulose acetate membrane electrophoresis. The monosaccharides composition of HSAP-2 was determined by RP-HPLC with pre-column 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization. The disaccharide composition of HSAP-2 was determined by mass spectrometry and a SAX-HPLC method, respectively, after digested by chondroitinase ABC. Results Two GAGs fractions were separated from Lapemis curtus skin, and the structural features of a pure GAGs fraction (HSAP-2) was preliminarily investigated. The disaccharide compositions of HSAP-2 contained four different disaccharides, namely, 4S, 6S and a few of 46S, 0S. HSAP-2 was consisted of glucuronic acid (GlcA) and acetyl galactosamine (GalNAc). It is a new GAGs that mainly contained hybrid dermatan sulfate (DS). Conclusion A novel dermatan sulfate was obtained from Lapemis curtus skin, and our data provide a basis for further research on the relationship of polysaccharide structure and biological activity form Lapemis curtus.