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

作　　者：白明月[1] 王清池 邓世明[4] 赵峡[1,2,3] BAI Ming-yue;WANG Qing-chi;DENG Shi-ming;ZHAO Xia(Key Laboratory of Marine Drugs, Ministry of Education, Shandong Provincial Key laboratory of Glycoscienee and Glycoengineering,School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003,China;Marine Biomedical Research Institute of Qingdao, Qingdao 266071,China;Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;Ocean College of Hainan Universitv. Haikou 570090, China)

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,医药学院,山东青岛266003 [2]青岛海洋生物医药研究院,山东青岛266071 [3]青岛海洋国家实验室海洋药物与生物制品功能实验室,山东青岛266237 [4]海南大学海洋学院,海南海口570228

出　　处：《中国海洋药物》2018年第1期7-14,共8页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金委/山东省政府联合基金项目(U1606403);青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02)资助

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)法分析其二糖组成。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种二糖,主要为单硫酸化二糖(4S和6S);单糖组成中含有艾杜糖醛酸(IdoA)、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。Objective To extract and isolate glycosaminoglycans（GAGs）fromLapemis 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,after digested by chondroitinase ABC.Results Two GAGs fractions were separated fromLapemis curtus skin,and the structural features of a pure GAGs fraction（HSAP-2）were preliminarily investigated.The disaccharide compositions of HSAP-2 contained four different disaccharides,mainly 4 Sand 6 S.HSAP-2 consisted of iduronic acid（IdoA）,glucuronic acid（GlcA）and acetyl galactosamine（GalNAc）.It was a new GAGs that mainly contained hybrid dermatan sulfate（DS）.Conclusion A novel dermatan sulfate was obtained from Lapemis curtus skin,and our data provided a basis for further research on the relationship of polysaccharide structure and biological activity of Lapemis curtus.

关 键 词：平颏海蛇 糖胺聚糖 分离纯化 二糖组成 

分 类 号：R931.74[医药卫生—生药学]