摘要
目的:从鲜地黄中提取并初步纯化α-半乳糖苷酶和β-葡萄糖苷酶。方法:以总酶活为考察指标,通过正交试验确定两种酶的最佳提取条件。以酶活力和蛋白质含量为考察指标,确定两步盐析法的硫酸铵饱和度。结果:提取温度和溶剂量对α-Gal的提取具有显著性影响,而提取溶剂对-βG lu的提取具有显著性影响,因此确定以3倍量(v/w)磷酸盐缓冲液,4℃提取4 h为两种酶的最佳提取条件。以30%和60%为两步盐析的最佳硫酸铵饱和点。结论:鲜地黄中存在α-半乳糖苷酶和β-葡萄糖苷酶,两步盐析法可以使二者得到初步纯化。
Objective： To extract and preliminarily purify α-galactosidase and β-glucosidase from the fresh roots of Rehmannia glntinosa. Methods： With the enzyme activity as a criterion, the best procedure of extraction was selected though orthogonal design method, and the desired saturation of ammonium sulfate in two-step salting-out was settled by gradient sedimentation of root extract according to enzyme activity and protein content. Results ： Temperature and solvent volume affect the extraction of α-galactosidase significantly, while solvent type to β-glucosidase. Therefore the procedure for extracting two enzymes was decided as mixing comminuted fresh ront with 3 times phosphate buffer, and placing the mixture in refrigeratory at 4℃ for 4 hours, and then obtaining the enzyme liquid by centrifuging at 4℃. 30% and 60% saturation was defined as the lower and upper point for two-step ammonium sulfate sahingout of tbe two enzymes. Conclusion： α-Galactosidase and β-glueosidase exist in the fresh roots, anti can be preliminarily purified through two-step salting-out.
出处
《中药材》
CAS
CSCD
北大核心
2006年第2期137-139,共3页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(No.30472189)
