| 白明月,王清池,邓世明,等.平颏海蛇皮中新型硫酸皮肤素的分离纯化和结构表征[J].中国海洋药物,2018,37(1):7-14. |
| 平颏海蛇皮中新型硫酸皮肤素的分离纯化和结构表征 |
| Isolation, purification and structural characterization of a novel dermatan sulfate from Lapemis curtus skin |
| 投稿时间：2017-07-26 修订日期：2017-09-24 |
| DOI： |
| 中文关键词: 平颏海蛇 糖胺聚糖 分离纯化 二糖组成 |
| English Keywords:Lapemis curtus,glycosaminoglycans,Isolation and purification,disaccharide composition |
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| 中文摘要: |
| 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。 |
| English Summary: |
| 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. |
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