摘要:
为了能高效快速的分离纯化桦褐孔菌子实体多糖,使用DEAE-52纤维素静态法,分别研究了静态吸附过程中多糖的样品浓度、吸附时间、吸附温度和吸附振荡转速等因素对多糖吸附量的影响及解吸过程中洗脱时间和洗脱液量对多糖解吸量的影响,确定了多糖分离纯化优化工艺条件。结果表明:多糖浓度为40 mg/mL时,在30 ℃,120 r/min转速下吸附处理90 min,此时DEAE-52纤维素对多糖吸附效果最好。而分别以12倍体积的去离子水洗脱90 min后,采用11倍体积的0.2 mol/L NaCl溶液洗脱60 min时,能达到最好的解吸效果,其中去离子水洗脱多糖浓度为0.56 mg/mL,0.2 mol/L NaCl洗脱多糖浓度为0.38 mg/mL。静态洗脱出的两种多糖组分均为分子量均一分布的多糖组分,与DEAE-52纤维素柱层析分离效果一致。因此采用DEAE-52纤维素静态法分离纯化桦褐孔菌子实体多糖是可行的。
Abstract:
In order to purify the polysaccharides efficiently and rapidly from the fruiting body of Inonotus obliquus,the DEAE-52 cellulose static absorption and desorption method was used. The polysaccharide concentration,adsorption time,adsorption temperature,rotate speed,elution time and eluent volume that influence the static adsorption and desorption of polysaccharide were selected and optimized. The optimal experimental conditions for static adsorption were determined as 40 mg/mL,90 min,30 ℃,120 r/min for sample concentration,adsorption time,adsorption temperature,and rotate speed,respectively. And the desorption effect was the best when eluted with 12 times volume of deionized water for 90 min and eluted with 11 times the volume of 0.2 mol/L NaCl solution for 60 min. Under the optimized conditions,the concentration of polysaccharide in deionized water and 0.2 mol/L NaCl elution polysaccharides were 0.56 mg/mL and 0.38 mg/mL,respectively. And,the two polysaccharides that were eluted were homogeneous distribution of polysaccharide components,which was consistent with the separation of DEAE-52 cellulose column chromatography in the previous experiment. Therefore,it is feasible to isolate and purify the polysaccharides of Inonotus obliquus by DEAE-52 cellulose static method.
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