甘露聚糖肽的结构鉴定

doi:10.7503/cjcu20160138

摘要/Abstract

摘要：

采用离子交换柱层析和凝胶柱层析从甘露聚糖肽原料中分离纯化得到4种均一糖肽MT1-A, MT1-B, MT2-A和MT2-B; 通过单糖组成分析、甲基化分析、氢核磁共振谱(¹H NMR)、红外光谱(IR)和氨基酸组成分析等手段对均一糖肽MT1-A, MT1-B和MT2-B的结构进行了鉴定. 结果表明, 3种均一糖肽的单糖组成、糖残基的连接方式与MT2-A相同, 但是分子量以及氨基酸总量有所不同. 测定了不同批次甘露聚糖肽原料的 ¹H NMR谱, 以MT2-A的 ¹H NMR谱图为标准谱, 以异头氢区域为鉴定区域, 通过计算不同批次甘露聚糖肽原料以及其它3种均一糖肽与MT2-A的 ¹H NMR谱的相关系数, 定量评价了不同批次甘露聚糖肽原料与4种均一糖肽的相似程度. 结果表明, 甘露聚糖肽是由糖链结构一致、分子量和氨基酸含量不同的几种均一糖肽构成的混合物.

关键词: 甘露聚糖肽, 多糖, 结构

Abstract:

DEAE-52 cellulose column and Sephacryl S-300 gel column were used to isolate and purify mannatide to give four homogeneous glycopeptides termed as MT1-A, MT1-B, MT2-A and MT2-B. The chemical structure of MT1-A, MT1-B and MT2-B were elucidated using monosaccharide composition analysis, methylation analysis, ¹H-nuclear magnetic resonance(¹H NMR) spectroscopy, infrared(IR) spectroscopy and amino acids analysis. It was concluded that the monosaccharide composition and connection type of sugar residues of MT1-A, MT1-B and MT2-A were the same as MT2-A reported before while the molecular weight and amino acid content of four homogeneous glycopeptides were slightly different. The ¹H NMR spectrum of MT2-A was regarded as the standard reference spectrum and the anomeric region of each spectrum(δ 4.89—5.90) was selected as the identification region. The ¹H NMR spectra of MT1-A, MT1-B, MT2-A and different batches of mannatide were compared with the standard reference spectrum, using the correlation coefficient to quantitatively calculate the similarity. The results showed that mannatide was a mixture of several kinds of homoge-neous glycopeptides with the same glycan structure but different molecular weight and amino acid content.

Key words: Mannatide, Polysaccharide, Structure

中图分类号:

O629.12

TrendMD:

沈艳红, 陈林枭, 张文清, 徐志珍, 宛燕飞, 张力, 夏玮. 甘露聚糖肽的结构鉴定. 高等学校化学学报, 2016, 37(6): 1088.

SHEN Yanhong, CHEN Linxiao, ZHANG Wenqing, XU Zhizhen, WAN Yanfei, ZHANG Li, XIA Wei. Structural Characterization of Mannatide^†. Chem. J. Chinese Universities, 2016, 37(6): 1088.

图/表 2

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Amino acid	Amino acids content/(g·100 g^-1)
Amino acid	MT1-A	MT1-B	MT2-A	MT2-B
Asparaginic acid	0.393	0.482	0.449	0.745
Threonine	0.655	0.780	0.870	1.480
Serine	0.672	1.392	0.700	1.921
Glutamic acid	0.350	0.529	0.389	0.972
Glycine	0.138	0.188	0.152	0.273
Alanine	0.477	1.036	0.470	1.254
Valine	0.219	0.287	0.258	0.534
Methionine	0.023	0.023	0.010	0.027
Isoleucine	0.087	0.130	0.112	0.207
Leucine	0.102	0.157	0.125	0.202
Tyrosine	0.078	0.097	0.102	0.165
Phenylalanine	0.067	0.090	0.0798	0.117
Histidine	0.037	0.056	0.037	0.055
Lysine	0.194	0.372	0.175	0.359
Arginine	0.047	0.087	0.036	0.066
Proline	0.340	0.444	0.447	0.734
Total content of amino acids	3.88	6.15	4.41	9.11

Amino acid	Amino acids content/(g·100 g^-1)
Amino acid	MT1-A	MT1-B	MT2-A	MT2-B
Asparaginic acid	0.393	0.482	0.449	0.745
Threonine	0.655	0.780	0.870	1.480
Serine	0.672	1.392	0.700	1.921
Glutamic acid	0.350	0.529	0.389	0.972
Glycine	0.138	0.188	0.152	0.273
Alanine	0.477	1.036	0.470	1.254
Valine	0.219	0.287	0.258	0.534
Methionine	0.023	0.023	0.010	0.027
Isoleucine	0.087	0.130	0.112	0.207
Leucine	0.102	0.157	0.125	0.202
Tyrosine	0.078	0.097	0.102	0.165
Phenylalanine	0.067	0.090	0.0798	0.117
Histidine	0.037	0.056	0.037	0.055
Lysine	0.194	0.372	0.175	0.359
Arginine	0.047	0.087	0.036	0.066
Proline	0.340	0.444	0.447	0.734
Total content of amino acids	3.88	6.15	4.41	9.11

Sample	Correlation efficient	Sample	Correlation efficient
Mannatide(G100102)	0.9937	Mannatide(140502)	0.9951
Mannatide(091202)	0.9942	MT1-A	0.9955
Mannatide(140401)	0.9761	MT1-B	0.9959
Mannatide(140501)	0.9783	MT2-B	0.9945

Sample	Correlation efficient	Sample	Correlation efficient
Mannatide(G100102)	0.9937	Mannatide(140502)	0.9951
Mannatide(091202)	0.9942	MT1-A	0.9955
Mannatide(140401)	0.9761	MT1-B	0.9959
Mannatide(140501)	0.9783	MT2-B	0.9945