Mechanism of the reaction catalyzed by dihydrofolate reductase from Escherichia coli: pH and deuterium isotope effects with NADPH as the variable substrate

Biochemistry ◽  
1988 ◽  
Vol 27 (15) ◽  
pp. 5499-5506 ◽  
Author(s):  
John F. Morrison ◽  
Stuart R. Stone
Keyword(s):  
Escherichia Coli ◽  
Dihydrofolate Reductase ◽  
Isotope Effects ◽  
Deuterium Isotope Effects ◽  
Variable Substrate ◽  
Mechanism Of The Reaction

The kinetics, isotope effects, and mechanism of the reaction of 2,2-di(4-nitrophenyl)-1,1,1-trifluoroethane with alkoxide bases in alcohol solvents

1985 ◽  
Vol 63 (3) ◽  
pp. 576-580 ◽  
Author(s):  
Arnold Jarczewski ◽  
Grzegorz Schroeder ◽  
Wlodzimierz Galezowski ◽  
Kenneth T. Leffek ◽  
Urszula Maciejewska
Keyword(s):  
Rate Constants ◽  
Isotope Effects ◽  
Activation Parameters ◽  
Tert Butanol ◽  
Deuterium Isotope Effects ◽  
Alcohol Solvents ◽  
Multistep Reaction ◽  
Mechanism Of The Reaction

The reaction between 2,2-di(4-nitrophenyl)-1,1,1-trifluoroethane and the alkoxide bases ŌCH3, ŌC2H5, ŌnC4H9, ŌCH(CH3)2, and ŌC(CH3)3 in their corresponding alcohol solvents is a multistep reaction with several intermediates: 2,2-di(4-nitrophenyl)-1,1-difluoro-1-alkoxyethane (A), 2,2-di(4-nitrophenyl)-1-fluoro-1-alkoxyethene (B), 2,2-di(4-nitrophenyl)-1,1-dialkoxyethene (C), 2,2-di(4-nitrophenyl)-1,1-difluoroethene (D), and 4,4′-dinitrobenzophene (E). Rate constants and activation parameters have been measured for the appearance of the two stable products B and C. The kinetic deuterium isotope effects for the appearance of B fell in the range of kH/kD = 1 to 2 at 25 °C for the primary and secondary alkoxides, whereas kH/kD = 5.4 at 30 °C for the appearance of D with tert-butoxide. Exchange experiments showed that H/D exchange took place between the substrate and solvent to the extent of 100% with methoxide, 50% with ethoxide and isopropoxide, and 0% with tert-butoxide. It is concluded the HF elimination from the substrate follows an (ElcB)R mechanism with methoxide/methanol, changing to (ElcB)I or E2 with tert-butoxide/tert-butanol.

Deuterium isotope effects on β-galactosidase formation by Escherichia coli

1967 ◽  
Vol 120 (2) ◽  
pp. 316-321 ◽  
Author(s):  
T.R. Henderson ◽  
Judy M. Dacus ◽  
H.L. Crespi ◽  
J.J. Katz
Keyword(s):  
Escherichia Coli ◽  
Isotope Effects ◽  
Deuterium Isotope Effects
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