中华皮肤科杂志 ›› 2000, Vol. 33 ›› Issue (5): 311-313.

• 论著 • 上一篇    下一篇

白念珠菌胞壁碱溶性β-葡聚糖的分离、纯化和分析

李岷, 陈沄, 邵凤, 沈永年, 吕桂霞, 陈伟, 郭宁如, 吴绍熙   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所, 南京210042
  • 收稿日期:1999-12-30 出版日期:2000-10-15 发布日期:2000-10-15

Isolation and Purification of the Alkali-soluble β-glucan from Cell Wall of Candida albicans

LI Min, CHEN Yun, SHAO Feng   

  1. Institute of Dermatolgoy, Chinese Academy of Medical Sciences&Peking Union Medical College, Nanjing 210042
  • Received:1999-12-30 Online:2000-10-15 Published:2000-10-15

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摘要： 目的 分离纯化白念珠菌胞壁碱溶性β-葡聚糖,并分析其物理化学性质和结构。方法与结果 白念珠菌标准株IDC1d经YEPD培养基28℃、18h培养,收集酵母相细胞,0.75mol/L NaOH液提取,乙醇沉淀,得粗多糖。采用红外光谱、苯酚-硫酸法、蒽酮-硫酸法、3,5-二硝基水杨酸法、考马斯亮蓝法定性分析粗多糖。粗多糖不含还原糖,但含有蛋白质。粗多糖经SephadexG-200凝胶柱层析纯化。纸层析、凝胶柱层析鉴定所得多糖的纯度,表明其为均一物质。凝胶柱层析测定相对分子质量约为89125.09。多糖经完全酸水解,硅胶薄层层析和气相色谱分析单糖组分,结果表明其为D-葡萄糖聚合成的多糖。红外光谱提示多糖糖苷键为β型。高碘酸氧化和Smith降解结果表明多糖包含1→3和1→6两种连接键,推测前者为主链键型,后者为支链键型。

关键词: 念珠菌,白色, β-葡聚糖

Abstract: Objective To isolate and purity the alkali-soluble β-glucan from the cell wall of Candida albicans and to analyze the physico-chemical properties and structure of it. Methods and Results After a standard strain of Candida albicans (IDC1d) was fermented on YEPD medium for 18 h at 28℃, yeast cells were digested with 3% NaOH solution and then crude glucan was extracted and precipitated by alcohol. Crude β-glucans was purified by Sephadex G-200 gel column chromatography. Paper chromatography and gel column chromatography were used to determine the purity of β-glucan, which was shown to be homogenous. The molecular weight of it was estimated to be 89 125.09 by gel column chromatography. β-glucan obtained from the cell wall of Candida albicans was composed of D-glucose only. Conclusion β-glucan is an important component of the cell wall of C.albicans.

Key words: Candida albicans, beta glucan