Plant flavokinase: affinity-chromatographic procedure for the purification of the enzyme from mung-bean (Phaseolus aureus) seeds and conformational changes on its interaction with orthophosphate

Abstract

Flavokinase was purified, for the first time from a plant source [mung bean (Phaseolus aureus)] by affinity chromatography in the presence of orthophosphate and by using C-8 ATP-agarose (ATP linked through the C-8 position to beaded agarose), Cibacron Blue and riboflavin-Sepharoses. An altered substrates-saturation pattern was observed in the presence of K₂HPO₄. The conformational changes of the enzyme in the presence of K₂HPO₄ were monitored by fluorescence spectroscopy. These results highlight the regulatory nature of this enzyme.