一种在多糖分离纯化过程中新的脱蛋白方法

目的　寻找一种在多糖 (香菇多糖 )分离纯化过程中适宜的脱蛋白方法。方法　以正交试验法为主。结果　通过正交试验法找到了一种在多糖 (香菇多糖 )分离纯化过程中新的脱蛋白方法。 5因素 3水平正交试验的结果表明 ,影响脱蛋白率 ( DPR)的主要因素是 B(正丁醇 )、次主要因素是 D(乙醇 ) ,而因素 C(甲醇 )和 E(氯化钠 )的影响则较小 ;影响 DPR的主次顺序为 B>D>A(氯仿 ) >E>C。 DPR随着正丁醇浓度的增加而变化很大 ,开始缓慢上升 ,然后迅速降低 ;随着乙醇浓度的增大 ,DPR逐渐降低 ;而当氯仿浓度增大时 ,DPR则开始上升 ,然后下降 ;当甲醇浓度增大时 ,DPR则变化很小。脱蛋白的最佳条件是 A2 B2 C1 D1 E2 ,即在氯仿 4 0 %、正丁醇 10 %、甲醇 2 %、乙醇4 0 %、氯化钠 0 .0 1%的条件下 ,杂蛋白的去除率最高。验证性试验结果表明 ,在此条件下 ,杂蛋白的去除率达99.98 %。通过反复试验 ,还找到了适用于工业化生产的包括香菇原料的蒸煮、煮沸样品的浓缩、粗多糖的沉淀和脱色、粗多糖的干燥等在内的原料预处理的最佳工艺。结论　这种新的脱蛋白方法具有工艺简单、无需贵重设备、加工成本低、容易放大等特点 ,特别适合于多糖类产品的精制或其他需要去除杂蛋白的单元操作

A novel approach to deproteinization in isolation and purification of lentinan from Lentinus edodes

Object Looking for a suitable approach to deproteinization in isolation and purification of lentinan from Lentinus edodes. Methods The orthogonal test was mainly used. Results A new approach was established for deproteinization in isolation and purification of a lentinan by orthogonal test. The results indicated that the major factor affecting the deproteinization rate (DPR) was B (2 butanol), the sub major factor D (ethanol); while C (methanol) and E (NaCl) had little influence. The influencing order for DPR was B>D>A (chloroform) >E>C. DPR varied greatly when the amount of 2 butanol varied, going up slowly first, then going down rapidly. DPR decreased gradually as the concentration of ethanol increased. DPR went up first, and then went down, when the content of chloroform increased. And DPR altered little when the concentration of methanol increased. The optimal condition for deproteinization was A 2B 2C 1D 1E 2, the best composition for the deproteinizing agent was 40% of chloroform, 10% of 2 butanol, 2% of methanol, 40% of ethanol, 0 01% of NaCl. Further proven experiments showed that DPR was up to 99 98% under the best condition. The optimal technology of pretreatment of the dried L. edodes, such as distilling of L. edodes, concentrating of boiled sample, precipitating and decolorizating of rough polysaccharides, drying of rough polysaccharides, which applied to massive production for industry, was also researched out through repeated trials. Conclusion The novel approach to deproteinization has the superiority of simple technology, needless high price apparatus, lower cost of processing, and easily amplifying. Thus, the technology of deproteinization is of great importance to the isolation and purification of fungi and other polysaccharides rich in proteins.