【作者】 田文敏；

【导师】 张丽萍；

【作者基本信息】 东北师范大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 黑盖木层孔菌（Phellinus nigricans）菌丝体经水提得菌丝体多糖PNW,通过正交试验确定菌丝体多糖的最优提取工艺为提取温度100℃,浸提4次,每次2h,浸提比（质量比）1:30,各因素影响大小的次序是:提取次数>浸提比>浸提时间。GC分析PNW的组成:由甘露糖和葡萄糖构成（摩尔比2.68:1.00）。经冻融分级、醇沉分级、脱蛋白、DEAE-纤维素柱层析和Sepharose CL-6B柱层析等方法,从PNW中纯化出均一多糖PNW1。HPLC检验为单一峰,平均分子量为33 KDa。GC分析PNW1由半乳糖、甘露糖、葡萄糖和少量的阿拉伯糖和岩藻糖组成,摩尔比为:8.77:6.44:3.26:1.00:1.35。对PNW1的结构分析采用了部分酸水解、高碘酸氧化、Smith降解、甲基化等化学分析方法及IR、GC、GC/MS、¹³C-NMR等仪器分析方法。确定了PNW1结构的重复单元。初步考察了PNW和PNW1对小鼠体外脾淋巴细胞增殖的影响,结果显示PNW和PNW1有增强脾淋巴细胞增殖的作用,PNW1的作用效果优于PNW。PNW和PNW1还可以促进ConA诱导的T淋巴细胞增殖作用和LPS诱导的B淋巴细胞增殖作用。更多还原

【Abstract】 The mycelia polysaccharides（PNW）were extracted from the mycelia of Phellinus nigricans using boiling water. The extracted condition of PNW was optimized by orthogonal experiment. The optimum condition was that the temperature 100℃, four times, two hours per times and the diffusion ratio 1:30.GC analysis indicated that the PNW was composed of Man and Glc with the molar ratio of 2.68: 1.00.PNW1 was purified from PNW using a combination of techniques, such as: ethanol fractional precipitation, deproteinization, DEAE cellulose column chromatography and Sepharose CL-6B column chromatography. HPLC analysis manifested that PNW1 was uniform fraction and the molecular weight was about 33 thousand Dalton. GC analysis indicated that the PNW1 was composed of Gal, Man, Glc, Ara and Xyl with the molar ratio of 8.77:6.44:3.26:1.00:1.35.The chemical and instrumental analysis methods such as : Partial acid hydrolysis, Periodate oxidation, Smith degradation, IR, GC, GC/MS and ¹³C-NMR were used on the structure analysis of PNW1.Then the repeated structural unit of PNW1 was determined.Initially investigated the effects on mouse leukomonocyte proliferation in vitro by PNW and PNW1. The results were that both PNW and PNW1 had significant effects on leukomonocyte proliferation and can promote the effects on the leukomonocyte proliferation stimulated by Con A or LPS. The effects of PNW1 on leukomonocyte proliferation were better than PNW.更多还原