DEAE-Sepharose Fast Flow分离纯化刺芹侧耳木质素降解酶

Purification of Ligninolytic Enzymes from Pleurotus eryngii by DEAE-Sepharose Fast Flow

[目的]为Pleurotus eryngii—Co60-7木质素降解酶的分离纯化和综合利用提供试验依据。[方法]采用DEAE—Sepharose^TM Fast Flow离子交换介质，分别考察缓冲液pH值、流速和洗脱方式等对刺芹侧耳木质素降解酶分离纯化的影响，确定了最佳分离纯化层析条件。[结果]DEAE-Sephalose^TM Fast Flow分离纯化Pleurotus eryngii-Co60-7木质素降解酶的最佳层析条件为：选择20mmol／L，pH值为5．0醋酸钠一醋酸缓冲体系，3ml／min的流速，进行分步洗脱（100、200～300和1000mmoL／L NaCl的三步洗脱），可较好地实现刺芹侧耳发酵液木质素降解酶初分，该纯化操作目标蛋白回收率达85％，纯化分离因素为2．71。[结论]该技术在分离纯化刺芹侧耳木质素降解酶上可行，具有潜在的工业应用价值。

[ Objective] The aim of the research was to provide the experimental basis for the purification and the comprehensive utilization of the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain. [ Method ] Purification of the Ligninolytic enzymes from Pleurotus eryngii-Co60- 7 strain was investigated by the anion-exchange media DEAE-SepharoseTM Fast Flow, by optimizing pH value of the buffer solution, flow rate and elution condition, and its optimal condition of the chromatography was confirmed. [ Results ] The optimal technical condition of DEAE- SepharoseTM Fast Flow for purification of the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain was as follows： the purification factor was 2.71 and Ligninolytic enzyme activity recovery was 85% through 20 mmol/L sodium acetate optimal buffer solution, pH value 5.0, 3 ml/min and 100,200 -300 mmol/L, 1 000 mmol/L three-step NaCl elution. [ Conclusion] The technique of separating the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain feasible and have potential applied value in dustry.