食品科学

• 基础研究 • 上一篇    下一篇

蛹虫草菌糠多糖的分离纯化及结构组成分析

张 颖,曾 艳,张丽姣,崔世茂,孙媛霞   

  1. 1.内蒙古农业大学农学院,内蒙古 呼和浩特 010010;2.中国科学院天津工业生物技术研究所,天津 300308
  • 出版日期:2014-07-15 发布日期:2014-07-18

Separation, Purification and Structural Analysis of Polysaccharides from Spent Mushroom Substrate of Cordyceps militaris

ZHANG Ying, ZENG Yan, ZHANG Li-jiao, CUI Shi-mao, SUN Yuan-xia   

  1. 1. Agricultural College, Inner Mongolia Agricultural University, Huhhot 010010, China;
    2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
  • Online:2014-07-15 Published:2014-07-18

摘要:

以蛹虫草菌糠为原料,利用纤维素酶酶解提取多糖,通过乙醇分级醇沉与Sevag法脱蛋白对所提多糖进行初级分离纯化,得到4 种多糖(P1、P2、P3、P4)。利用凝胶色谱、气相色谱-质谱联用技术(gas chromatographymassspectrometry,GC-MS)对这4 种多糖的分子质量分布与单糖组成进行分析;进一步利用Superdex 200凝胶柱层析对P2进行纯化精制,通过高碘酸钠氧化、Smith降解、红外光谱和核磁共振(nuclear magnetic resonance,NMR)技术表征了精多糖P2的一级结构。结果表明:乙醇分级沉淀与Sevag脱蛋白方法的结合能满足蛹虫草菌糠多糖初级纯化的要求,得到了较纯的P2、P3、P4组分,分子质量分别为35.9、9.4、3.7 kD;其中P2是由葡萄糖组成的葡聚糖,其主链结构主要为α-(1→4)吡喃葡聚糖。P3和P4是由甘露糖、葡萄糖和半乳糖组成的杂多糖,且单糖组成均以葡萄糖为主。

关键词: 蛹虫草, 菌糠多糖, 分离纯化, 结构分析

Abstract:

Four polysaccharides (P1, P2, P3, and P4) were prepared from the spent mushroom substrate of Cordyceps
militaris after the extraction with cellulase, fractional precipitation with ethanol and deproteinization with Sevag regent.
Their molecular weight distribution and monosaccharide composition were analyzed by gel chromatography and gas
chromatography-mass spectrometry (GC-MS). The polysaccharide P2 was further purified by Superdex 200 gel-filtration
column and its primary structure was studied by sodium periodate oxidation, Smith degradation, infrared spectroscopy as
well as nuclear magnetic resonance. The results showed that the fractional precipitation by ethanol in combination with
deproteinization by Sevag regent could provide preliminary purification for the polysaccharides, resulting in 3 relatively
pure polysaccharides (P2, P3, and P4) with molecular weight of 35.9, 9.4 and 3.7 kD, respectively. The structural analysis
demonstrated that P2 was a glucan with a chain mainly comprised of α-(1→4) pyran glucan. Both P3 and P4 were
heteropolysaccharides consisting of mannose, galactose and glucose with glucose being the main monosaccharide.

Key words: Cordyceps militaris, polysaccharides from spent mushroom substrate, separation and purification, structural analysis