The preparation of an enzyme associated with aflatoxin biosynthesis by affinity chromatography.
An affinity matrix for the purification of norsolorinic acid dehydrogenase, an enzyme involved in aflatoxin biosynthesis, was prepared by coupling norsolorinic acid to an agarose gel. This matrix was found to be ineffective in isolating active enzyme, and was therefore modified by methylation, using diazomethane. The methylated matrix produced a one-step purification of the enzyme from a crude homogenate, resulting in a 138-fold purification. The active isolate was found to contain one major and two minor bands upon nondenaturing electrophoresis, and all the norsolorinic acid dehydrogenase activity was associated with the major band. It was concluded that the matrix exhibited true affinity for the enzyme, and that affinity chromatography was a valuable approach to isolating other secondary metabolic enzymes involved in the biosynthesis of the aflatoxins.
Department of Biochemistry, University of Natal, Pietermaritzburg, South Africa.
A A
AA
Chuturgoon
M F
MF
Dutton
R K
RK
Berry
D000348:Aflatoxins / Q000096:biosynthesis; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D001230:Aspergillus / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods; D005656:Fungal Proteins; D007202:Indicators and Reagents; D064430:NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases
D000348:Aflatoxins; D005656:Fungal Proteins; D007202:Indicators and Reagents; D000429:Alcohol Oxidoreductases; D064430:NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases; C062609:norsolorinic acid ketoreductase