猴头菇多糖粗提液的壳聚糖法脱蛋白及分离纯化研究

Protein removal and purification by chitosan method of Hericium erincaceus polysaccharide extract

研究了料液比(v/v)、p H、时间及处理次数对猴头菇多糖粗提液壳聚糖法脱蛋白的影响,得到了壳聚糖法脱蛋白的最优工艺,并通过柱层析对猴头菇多糖进行了分离纯化。优化的壳聚糖法脱蛋白工艺为:料液比50∶1,p H为粗提液本身的p H,4℃条件下静置2 h,处理次数为1次;此时蛋白质去除率为60.11%±0.47%,多糖损失率为11.16%±0.48%,壳聚糖残留率为1.01%±0.005%,蛋白质去除率比Sevage法提高11.67%,且多糖损失率降低了80.87%。猴头菇粗多糖经DEAE Sepharose Fast Flow柱层析分离得到两种多糖HEP-Ⅰ和HEP-Ⅱ,Sephadex G-75柱层析、紫外检测得出HEP-Ⅰ和HEP-Ⅱ均为成分均一的多糖,红外检测得出HEP-Ⅰ为同时含有吡喃环和呋喃环的糖类物质,而HEP-Ⅱ为含有吡喃环的糖类物质。壳聚糖法脱蛋白工艺合理可行,分离纯化后的猴头菇多糖成分均一。

Effects of solid-liquid ratio（v/v）, pH, time and frequency on protein removal from Hericium erincaceus poiysaccharide extract by chitosan method were studied,and the optimum conditions were obtained,the Hericium erincaceus polysaccharide was isolated and purified by column chromatography. The results showed that the optimum conditions were the ratio of extract to chitosan solvent（v/v） was 50：1, original pH, and time of 2 h for a time.Under such condition, the removal rate of protein reached about 60.11% ± 0.47%, which was higher than Sevage method by 11.67% and the loss rate of polysaccharides was decreased to about 11.16% ±0.48%,which was lower than Sevage method by 80.87% ,and the residual of chitosan was detected about 1.01 %± 0.005%.Two kinds of polysaccharide, HEP- Ⅰ and HEP- Ⅲ, were isolated and purified from the crude Hericium erinaceus polysaccharide by DEAE Sepharose Fast Flow column chromatography.Sephadex G-75 column chromatography, UV spectrophotometer,and Fourier transform infrared spectroscopy were used to analyze HEP- Ⅰ and HEP- Ⅱ. The results showed that both of HEP- Ⅰ and HEP- Ⅱ were in unique form.And HEP- Ⅰ contain carbohydrate pyran ring and furan ring,while HEP- Ⅱ only contain carbohydrate pyran ring.Chitosan method was reasonably practicable,and Hericium erinaceus polysaccharide was in high purity after column chromatography.