固定化亲和层析分离灵芝发酵液蛋白酶A抑制剂

Screening the inhibitors for PrA from Ganoderma lucidum by immobilized enzyme affinity chromatography

建立了一种固定化亲和层析高效分离筛选蛋白酶A抑制剂的方法,在pH为4.0,戊二醛终浓度为0.6%,给酶量为40mg/0.2g壳聚糖的条件下,固定化酶的回收率较高,达45.9%;利用此固定化蛋白酶A(PrA)亲和柱一步分离灵芝发酵全粉抽提液中的PrA抑制剂,所得抑制剂与原两步层析法分离的PrA抑制剂GLPAI在分子量、糖与蛋白质比例及抑制特性上都较为相近。

An effective method screening for the PrA by immobilized enzyme nhibitor of chromatography was set up, The affinity optimal conditions for immobilizing reaction were as follows： pH of reaction was 4.0;the concentration of the glutaraldehyde was 0.6% and the addition of PrA was 40mg/0.2g chitosan, The yield of immobilized PrA was 45.9% under the optimal conditions. The inhibitor purified by affinity chromatography and GLPA1 purified by two step chromatography were the same because they showed little difference in the molecular weight and the proportion of protein to glucose.