Screening method for large numbers of dye-adsorbents for enzyme purification.
A method is described by means of which 96 different dye-adsorbents can be tested simultaneously for their ability to bind enzymes and to test their biospecific elution. Small amounts of cell-free extract are applied to dye-adsorbents which are packed in a 96-well transplate cartridge. After biospecific elution, the amount of the eluted enzyme is tested in a microtitre plate assay. The method is illustrated by the purification of glycerol dehydrogenase (E.C. 1.1.1.72), 6-phosphogluconate dehydrogenase (E.C. 1.1.1.44) and glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) from the hyphal fungus Aspergillus nidulans.
Department of Genetics, Agricultural University, HA Wageningen, The Netherlands.
D H
DH
Hondmann
J
J
Visser
D001233:Aspergillus nidulans / Q000201:enzymology* / Q000254:growth & development; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003124:Colorimetry; D004396:Coloring Agents; D004798:Enzymes / Q000302:isolation & purification*; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D010734:Phosphogluconate Dehydrogenase / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet; D013401:Sugar Alcohol Dehydrogenases / Q000302:isolation & purification
D004396:Coloring Agents; D004798:Enzymes; D011506:Proteins; D013401:Sugar Alcohol Dehydrogenases; D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; C020672:glycerol dehydrogenase