[Extraction and salting-out purification of alpha-galactosidase and beta-glucosidase from fresh roots of Rehmannia glutinosa].
OBJECTIVE
To extract and preliminarily purify alpha-galactosidase and beta-glucosidase from the fresh roots of Rehmannia glutinosa.


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 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.


CONCLUSIONS
alpha-Galactosidase and beta-glucosidase exist in the fresh roots, and can be preliminarily purified through two-step salting-out.
Zhao
Yu
Y
School of Pharmaceutical Sciences, Shandong University, Jinan 250012.
Wen
Xue-sen
XS
Cui
Jing
J
Wu
Wei-hong
WH
D008970:Molecular Weight; D018517:Plant Roots / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*; D032267:Rehmannia / Q000737:chemistry*; D013678:Technology, Pharmaceutical / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D000519:alpha-Galactosidase / Q000302:isolation & purification* / Q000378:metabolism; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D001617:beta-Glucosidase; D000519:alpha-Galactosidase