Escherichia coli glyoxalate carboligase. Properties and reconstitution with 5-deazaFAD and 1,5-dihydrodeazaFADH2.
Glyoxalate carboligase (EC 4.1.1.47) has been purified to electrophoretic homogeneity from Escherichia coli. The enzyme was found to be a dimer of subunits of identical molecular weight of 68,000. Resolution of the holoenzyme into apoenzyme and FAD led to a dissociation of the dimer into monomers. The apoenzyme could be reconsitituted to full catalytic activity with FAD or the flavin coenzyme analogue 5-deazaFAD. Reconstitution of the apoenzyme with the reduced flavin analogue 1,5-dihydro-5-deazaFADH2 led to the recovery of 50% of enzymatic activity. The reconstitution of apoglyoxalate carboligase with all three coenzymes followed Michaelis-Menten kinetics with Km values of 0.25, 0.74, and 0.72 muM for FAD deazaFAD, and deazaFADH2, respectively.
D001665:Binding Sites; D002262:Carboxy-Lyases / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives* / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011485:Protein Binding; D013053:Spectrophotometry; D013056:Spectrophotometry, Ultraviolet
D046911:Macromolecular Substances; D005182:Flavin-Adenine Dinucleotide; D002262:Carboxy-Lyases