PMID: 16617782Apr 19, 2006Paper

Extraction and salting-out purification of alpha-galactosidase and beta-glucosidase from fresh roots of Rehmannia glutinosa

Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials
Yu ZhaoWei-hong Wu

Abstract

To extract and preliminarily purify alpha-galactosidase and beta-glucosidase from the fresh roots of Rehmannia glutinosa. 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. Temperature and solvent volume affect the extraction of alpha-galactosidase significantly, while solvent type to beta-glucosidase. Therefore the procedure for extracting two enzymes was decided as mixing comminuted fresh root with 3 times phosphate buffer, and placing the mixture in refrigeratory at 4 degrees C for 4 hours, and then obtaining the enzyme liquid by centrifuging at 4 degrees C. 30% and 60% saturation was defined as the lower and upper point for two-step ammonium sulfate salting-out of the two enzymes. alpha-Galactosidase and beta-glucosidase exist in the fresh roots, and can be preliminarily purified through two-step salting-out.

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