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引用本文:苏倡,刘艳,蒋鑫,苏秀榕.基于FT-IR、GC及核磁共振的仿刺参(Apostichopus japonicus)消化道多糖结构解析研究.海洋与湖沼,2019,50(4):886-893.
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基于FT-IR、GC及核磁共振的仿刺参(Apostichopus japonicus)消化道多糖结构解析研究
苏倡1, 刘艳1, 蒋鑫2, 苏秀榕2
1.宁波职业技术学院 乙烯工程副产物高质化利用浙江省应用技术协同创新中心 宁波 315800;2.宁波大学海洋学院 宁波 315211
摘要:
仿刺参(Apostichopus japonicus)因其高营养价值并富含多种生物活性成分,具有重要的经济价值。为了充分利用仿刺参加工下脚料,提高养殖经济效益,本文采用复合蛋白酶解法提取仿刺参消化道多糖,研究了酶解温度、时间、pH及加酶量对仿刺参消化道多糖得率的影响,并通过正交实验对仿刺参消化道多糖提取工艺进行优化,利用DEAE-sepharose Fast Flow层析柱对多糖进行纯化,通过傅里叶红外光谱(FT-IR)、气相色谱(GC)及核磁共振(13C,1H谱)对多糖的结构及组成进行分析。结果表明,复合蛋白酶在pH 7,加酶量1.2×104 U/g,温度60℃酶解6h;利用Sevage法脱蛋白提取得到的仿刺参消化道多糖纯度均一,效果最佳。通过FT-IR结果判断其为糖类化合物,含有β型葡萄毗喃糖苷键;利用气相色谱确定该多糖主要由鼠李糖、岩藻糖、阿拉伯糖、甘露糖、葡萄糖和半乳糖六种单糖组成,其摩尔比为1︰32.86︰14.56︰1.66︰0.78︰10.43,其中岩藻糖的含量最高;核磁共振综合分析结果进一步证实仿刺参消化道多糖含有β型毗喃已糖,且主要是以l,4-糖苷键相连接。
关键词:  仿刺参(Apostichopus japonicus)消化道  复合蛋白酶  正交实验  多糖制备  结构解析
DOI:10.11693/hyhz20190300063
分类号:Q539
基金项目:浙江省访问工程师资助项目,FG2017055号;国家农业科技成果转化项目,2007GB2C220号。
PREPARATION AND STRUCTURE ANALYSIS OF APOSTICHOPUS JAPONICUS DIGESTIVE TRACT
SU Chang1, LIU Yan1, JIANG Xin2, SU Xiu-Rong2
1.Zhejiang Collaborative Innovation Center for High Value Utilization of Byproducts from Ethylene Project, Ningbo Polytechnic College, Ningbo 315800, China;2.School of Marine Science, Ningbo University, Ningbo 315211, China
Abstract:
In this study, polysaccharide in Apostichopus japonicus digestive tract was extracted in complex protease hydrolysis extraction. The effects of extraction temperature, extraction time, pH, and enzyme amount on the yields of polysaccharides were investigated. The extraction process of polysaccharide was optimized through the orthogonal experimental. The structure and component of A. japonicus digestive tract polysaccharide was characterized using DEAE-sepharose Fast Flow, FT-IR, GC, and 13C, 1H nuclear magnetic resonance. The best extract conditions for A. japonicus polysaccharide were at 60℃ in enzyme amount of 1.2×104 U/g at pH 7.0 for treatment 6h and using Sevage method to todeproteinization. From FT-IR, it was a polysaccharide and contained β-D-glucopyranoside bond. The analysis of monosaccharide composition in the polysaccharide by GC revealed that it was composed of rhamnose, fucose, arabinose, mannose, glucose, and galactose in the molar ratios of 1︰32.86︰14.56︰1.66︰0.78︰10.43. Nuclear magnetic resonance analysis result shows that the structure of polysaccharide was mainly composed with 1→4 glycosidic bond.
Key words:  Apostichopus japonicus digestive tract  compound protease  orthogonal experiment  polysaccharide preparation  structural analysis
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