In metabolic networks, metabolites are usually present in great excess over the enzymes that catalyze their interconversion, and describing the rates of these reactions by using the Michaelis-Menten rate law is perfectly valid. This rate law assumes that the concentration of enzyme-substrate complex (C) is much less than the free substrate concentration (S0). However, in protein interaction net...

The or-chymotrypsin-catalyzed hydrolysis of p-nitrophenyl acetate has been studied at pH 7 and 25’ in isopropyl alcoholwater and dioxane-water mixtures. It was found that the initial reaction in which the enzyme is acylated follows second order kinetics rather than Michaelis-Menten kinetics as previously reported. The second order rate constant for this reaction is markedly decreased by additio...

Recent interest in modeling biochemical networks raises questions about the relationship between often complex mathematical models and familiar arithmetic concepts from classical enzymology, and also about connections between modeling and experimental data. This review addresses both topics by familiarizing readers with key concepts (and terminology) in the construction, validation, and applica...

The enzymatic processes according Michaelis-Menten kinetics have been studied from various approaches to describe the inhibition state. Proposals for inhibition were compared from a generic process, where kinetic constants have received unitary values, and the numeric value of the concentration of substrate was ten (10) times higher than the numerical value of the concentration of enzyme. For e...

Stereoselective transport of methotrexate (L-amethopterin, L-MTX) and its antipode (D-amethopterin, D-MTX) by the proton-coupled folate transporter (PCFT) was examined using PCFT-expressing HEK293 cells (PCFT-HEK293 cells). Uptake of both L-MTX and D-MTX was pH-dependent and decreased with an increase in the extracellular pH from 5.0 to 7.4. The initial uptake rate of L-MTX into PCFT-HEK293 cel...

When sycamore cells were suspended in basal medium containing choline, the latter was taken up by the cells very rapidly. A facilitated diffusion system appertained at low concentrations of choline and exhibited Michaelis-Menten kinetics. At higher choline concentrations simple diffusion appeared to be the principal mode of uptake. Addition of choline to the perfusate of compressed sycamore cel...

An in vitro technique for perfusion of the intestinal vasculature and lumen was developed and used to measure calcium (Ca) fluxes across the intestinal mucosa of the marine teleost, the Atlantic cod {Gadus morhua). Saturable and nonsaturable components of the calcium influx and efflux were determined. The calcium influx had one passive component and one saturable component, following Michaelis-...

DL-Malic acid-grown cells of the yeast Hansenula anomala formed a saturable transport system that mediated accumulative transport of L-malic acid with the following kinetic parameters at pH 5.0: Vmax, 0.20 nmol.s-1.mg (dry weight)-1; Km, 0.076 mM L-malate. Uptake of malic acid was accompanied by proton disappearance from the external medium with rates that followed Michaelis-Menten kinetics as ...

We have measured the uptake of arginine into vacuolar membrane vesicles from Neurospora crassa. Arginine transport was found to be dependent on ATP hydrolysis, Mg2+, time, and vesicle protein with transported arginine remaining unmodified after entry into the vesicles. The Mg2+ concentration required for optimal arginine transport varied with the ATP concentration so that maximal transport occu...

This paper describes a microanalytical method for determining enzyme kinetics using a continuous-flow microfluidic system. The analysis is carried out by immobilizing the enzyme on microbeads, packing the microbeads into a chip-based microreactor (volume approximately 1.0 nL), and flowing the substrate over the packed bed. Data were analyzed using the Lilly-Hornby equation and compared to value...