[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...[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。展开更多
[Objective] This study aimed to establish an efficient process for separation of phycoerythrin by using Q Sepharose Fast Flow resin and verity its feasibility for scale-up. [Method] Elution gradient, sample volume and...[Objective] This study aimed to establish an efficient process for separation of phycoerythrin by using Q Sepharose Fast Flow resin and verity its feasibility for scale-up. [Method] Elution gradient, sample volume and flow rate were optimized to determine the optimal separation condition, under which the scale-up process was verified. [Result] The optimal condition for separation of phycoerythrin by using Q Sepharose FF resin was investigated： 30 ml of laver extract was loaded to the Q Sepharose FF column with a bed volume of 8 ml; subsequently, the column was stepwise eluted with 0-0.10-0.35-1.00 mol/L NaCI solution （pH 6.0） at a constant flow rate of 1 ml/min; the elution peak under 0.35 mol/L NaCI solution was collected, and the recovery rate and purity coefficient （A565/A280） of phycoerythrin were determined as 44.3 and 1.15, respectively. Based on the established process, 75 ml of phycoerythrin extract was loaded to the Q Sepharose FF column with a bed volume of 20 ml for separation, while no significant variation was observed in the separation result. [Conclusion] Phycoerythrin can be well separated from laver extract by using Q Sepharose FF resin and the process is feasible for scale-up.展开更多
目的:优选分离冬凌草多糖的方法,探究不同分离方法对冬凌草多糖分离效果的影响。方法:分别以DEAE-Sepharose Fast Flow(Cl-型)弱极性阴离子交换层析法、乙醇分级醇析法对冬凌草粗多糖进行分离纯化,以不同分离组分中多糖含量及其免疫活...目的:优选分离冬凌草多糖的方法,探究不同分离方法对冬凌草多糖分离效果的影响。方法:分别以DEAE-Sepharose Fast Flow(Cl-型)弱极性阴离子交换层析法、乙醇分级醇析法对冬凌草粗多糖进行分离纯化,以不同分离组分中多糖含量及其免疫活性比较优选冬凌草粗多糖的分离工艺。结果:DEAE-Sepharose Fast Flow(Cl-型)弱极性阴离子交换层析法分离得到5个组分,通过乙醇分级醇沉初步得到4个冬凌草多糖组分,前者分离产品纯度较高,而后者产品在体外小鼠脾淋巴T细胞的增殖实验中表现出较强生物活性。结论:为进一步研究冬凌草多糖的生物活性,可选用乙醇分级醇沉法对冬凌草多糖进行分离纯化。展开更多
文摘[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。
基金Supported by National Natural Science Foundation of China(51143012)Natural Science Foundation of Shandong Province(ZR2009BM006)~~
文摘[Objective] This study aimed to establish an efficient process for separation of phycoerythrin by using Q Sepharose Fast Flow resin and verity its feasibility for scale-up. [Method] Elution gradient, sample volume and flow rate were optimized to determine the optimal separation condition, under which the scale-up process was verified. [Result] The optimal condition for separation of phycoerythrin by using Q Sepharose FF resin was investigated： 30 ml of laver extract was loaded to the Q Sepharose FF column with a bed volume of 8 ml; subsequently, the column was stepwise eluted with 0-0.10-0.35-1.00 mol/L NaCI solution （pH 6.0） at a constant flow rate of 1 ml/min; the elution peak under 0.35 mol/L NaCI solution was collected, and the recovery rate and purity coefficient （A565/A280） of phycoerythrin were determined as 44.3 and 1.15, respectively. Based on the established process, 75 ml of phycoerythrin extract was loaded to the Q Sepharose FF column with a bed volume of 20 ml for separation, while no significant variation was observed in the separation result. [Conclusion] Phycoerythrin can be well separated from laver extract by using Q Sepharose FF resin and the process is feasible for scale-up.
文摘目的:优选分离冬凌草多糖的方法,探究不同分离方法对冬凌草多糖分离效果的影响。方法:分别以DEAE-Sepharose Fast Flow(Cl-型)弱极性阴离子交换层析法、乙醇分级醇析法对冬凌草粗多糖进行分离纯化,以不同分离组分中多糖含量及其免疫活性比较优选冬凌草粗多糖的分离工艺。结果:DEAE-Sepharose Fast Flow(Cl-型)弱极性阴离子交换层析法分离得到5个组分,通过乙醇分级醇沉初步得到4个冬凌草多糖组分,前者分离产品纯度较高,而后者产品在体外小鼠脾淋巴T细胞的增殖实验中表现出较强生物活性。结论:为进一步研究冬凌草多糖的生物活性,可选用乙醇分级醇沉法对冬凌草多糖进行分离纯化。
