In situ and in vitro kinetics of phenol hydroxylase.
The half saturation constant for phenol was much lower with phenol hydroxylase in situ than with the purified enzyme, whereas the constant for NADPH was higher. In both cases, the linearized plots of the Michaelis-Menten equation were biphasic and the half saturation constants for all phenolic substrates were several times lower, when the phenol was added to the assay medium before NADPH, than when NADPH was added first. There was a similar, but much smaller, effect on the half saturation constants for NADPH. The V-values were not affected by the order of addition. The results suggest slow conformational changes in the enzyme during the overall reaction, which seem even slower, when the enzyme is measured in situ.
D002396:Catechols / Q000378:metabolism; D007700:Kinetics; D008433:Mathematics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010636:Phenols / Q000378:metabolism; D012118:Resorcinols / Q000378:metabolism; D013329:Structure-Activity Relationship
D002396:Catechols; D010636:Phenols; D012118:Resorcinols; D009249:NADP; D006899:Mixed Function Oxygenases; C022336:phenol 2-monooxygenase; C034221:catechol; C031389:resorcinol