Steady-state kinetic properties of sorbitol dehydrogenase from chicken liver.
The steady-state kinetic properties of partially purified chicken liver sorbitol dehydrogenase (SDH) were determined spectrophotometrically at 25 degrees C, in 50 mM 3-(N-morpholino)propanesulfonic acid (MOPS) buffer, pH 8.0. In the sorbitol-to-fructose direction, analysis was based on initial rate data obtained at [NAD(+)](o)=0.1-0.4 mM and [sorbitol](o)=1.25-10 mM. The reverse process was analyzed by recording progress curves for NADH consumption, starting with [NADH](o)=0.2 mM and [fructose](o)=66.7-267 mM. The kinetics conformed to an ordered sequential model, with the cofactors adding first. The steady-state parameters in the forward direction, K(NAD(+)), K(iNAD(+)) and K(sorbitol), were found to be 210+/-62 muM, 220+/-69 microM and 3.2+/-0.54 mM, respectively. The corresponding parameters in the reverse direction were K(NADH)=240+/-58 microM, K(iNADH)=10+/-2.8 microM and K(fructose)=1000+/-140 mM. The results indicated a close parallelism with human SDH, yet up to 40-fold differences were observed when compared to related reports on other mammalian species. The structural and adaptive bases of the variation in substrate and cofactor affinities need to be accounted for.
Karacaoğlan
Volkan
V
Department of Biochemistry, School of Pharmacy, Hacettepe University, 06100 Ankara, Turkey.
Ozer
Inci
I
D000818:Animals; D002645:Chickens; D005632:Fructose / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D007064:L-Iditol 2-Dehydrogenase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D013012:Sorbitol / Q000378:metabolism
D005632:Fructose; D013012:Sorbitol; D007064:L-Iditol 2-Dehydrogenase