雷蘑胞外多糖分离提取工艺优化研究

Technical optimization on separation and purification of extracellular polysaccharide from Clitocybe gigantean

优化了雷蘑(Clitocybe gigantean)AS 5.105深层发酵胞外多糖的分离提取工艺条件,以醇沉物脱蛋白后的纯多糖为研究对象,胞外多糖得率EPGR(Exopolysaccharide gain rate)和脱蛋白率DPR(Deproteinized rate)为评价指标,采用Mixture design和3因素3水平Box-Behnken统计学实验设计方法,确定了胞外多糖得率及Sevag法脱蛋白的最优条件。结果表明,在乙醇终浓度为77.5%～84.2%、醇沉温度为4.2℃～1.3℃时,于18.1h～24h内胞外多糖得率92.79%;在氯仿、正丁醇和透析液分别为34%、12%和54%的配比条件下,脱蛋白率预测值为98.64%。通过对脱蛋白率模型方程求解可知,脱蛋白次数、每次脱蛋白时间以及脱蛋白温度分别为8次、23.85min和14.27℃时,脱蛋白率最大可达96.99%。

In order to optimize the condition of extraction and separation of extracellular polysaccharide from Clitocybe gigantean AS 5.105, with pure polysaccharide as research objective, with the exopolysaccharide gain rate （EPGR） and deproteinized rate （DPR） as evaluation index, the optimal conditions to obtain EPGR an DPR were determined by Mixture design and three-level three-factor Box-Behnken design. The results showed that the maximum EPRG （92.79%） could be obtained in 18.1-24 h with the final ethanol concentration as 77.5%-84.2% and precipitation temperature as 4.2℃-11.3℃. The predicted DPR of 98.64% was obtained with miture of chloroform, butanol and dialysis solutions at the ratio of 34：12：54. Based on DPR model equation, the maximum predicted DPR of 96.99% was reached with the deproteinized times as 8, holding time as 23.85 min and deproteinized temperature as 14.27℃.