Deviation from Michaelis-Menten kinetics for fumarase

A study of the steady-state kinetics of fumarase over an extended concentration range, using novel methods of analysis, reveals an initial-rate equation of at least fourth degree for malate as substrate at pH 7.0, with no kinetically significant dead-end complex formation even up to concentrations of 100 mM. In the absence of demonstrable enzyme-aggregation phenomena, this is interpreted as indicating co-operative effects overlooked previously, although a mixture of isoenzymes, each individually of high degree and giving a complex curve, may be a contributing factor.

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Steady-state kinetics of thyroid peroxidase. evidence for a high degree rate equation using the f statistic

International Journal of Biochemistry ◽

10.1016/0020-711x(83)90239-2 ◽

1983 ◽

Vol 15 (9) ◽

pp. 1195-1200

Author(s):

Francisco Solano-Muñoz ◽

JoséL. Iborra ◽

JoséA. Lozano ◽

William G. Bardsley

Keyword(s):

Steady State ◽

Rate Equation ◽

Thyroid Peroxidase ◽

Steady State Kinetics ◽

High Degree ◽

Kinetics Of

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Steady-state kinetic analysis of aldehyde dehydrogenase from human erythrocytes

Biochemical Journal ◽

10.1042/bj2870145 ◽

1992 ◽

Vol 287 (1) ◽

pp. 145-150 ◽

Cited By ~ 8

Author(s):

G T M Henehan ◽

K F Tipton

Keyword(s):

Steady State ◽

Aldehyde Dehydrogenase ◽

Initial Rate ◽

Substrate Inhibition ◽

Human Erythrocytes ◽

Steady State Kinetics ◽

Steady State Kinetic ◽

Sequential Mechanism ◽

Kinetics Of

The steady-state kinetics of purified cytoplasmic aldehyde dehydrogenase (EC 1.2.1.3) from human erythrocytes have been studied at 37 degrees C. Previous studies of the enzyme from several mammalian sources, which used a lower assay temperature, have been difficult to interpret because of the substrate activation by acetaldehyde which led to complex kinetic behaviour. At 37 degrees C the initial-rate data do not depart significantly from Michaelis-Menten kinetics. Studies of the variation of initial rates as a function of the concentrations of both substrates and studies of the inhibition by NADH were consistent with a sequential mechanism being followed. High-substrate inhibition by acetaldehyde was competitive with respect to NAD+. The enzyme was not inhibited by the product acetate and thus the results of these studies, although consistent with an ordered mechanism in which NAD+ was the first substrate to bind, were inconclusive. That such a mechanism was followed was confirmed by determination of the initial-rate behaviour in the presence of acetaldehyde and glycolaldehyde as alternative substrates. When the reciprocal of the initial rate of NADH formation was plotted against the acetaldehyde concentration at a series of fixed ratios between that substrate and glycolaldehyde, a linear ‘mixed inhibition’ pattern was obtained, confirming the mechanism to be ordered with NAD+ being the leading substrate and with kinetically significant ternary complex-formation.

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Thermodynamic and detailed kinetic study of the formation of orthophthalaldehyde-agmatine complex by fluorescence intensities

Journal of Analytical Science & Technology ◽

10.1186/s40543-020-00238-2 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Khémesse Kital ◽

Moumouny Traoré ◽

Diégane Sarr ◽

Moussa Mbaye ◽

Mame Diabou Gaye Seye ◽

...

Keyword(s):

Complex Formation ◽

Thermodynamic Parameters ◽

Initial Rate ◽

Reaction Medium ◽

Standard Entropy ◽

Step Size ◽

Formation Reaction ◽

Different Temperatures ◽

Kinetics Of ◽

Complex Formation Process

Abstract The aim of this work is to determine the thermodynamic parameters and the kinetics of complex formation between orthophthalaldehyde (OPA) and agmatine (AGM) in an alkaline medium (pH 13). Firstly, the association constant (Ka) between orthophthalaldehyde and agmatine was determined at different temperatures (between 298 K and 338 K) with a step size of 10 K. Secondly, the thermodynamic parameters such as standard enthalpy (ΔH°), standard entropy (ΔS°),and Gibbs energy (∆G) were calculated, where a positive value of ΔH° (+45.50 kJ/mol) was found, which shows that the reaction is endothermic. In addition, the low value of ΔS°(+0.24 kJ/mol) indicates a slight increase in the disorder in the reaction medium. Furthermore, the negative values of ΔG between −35.62 kJ/mol and −26.02 kJ/mol show that the complex formation process is spontaneous. Finally, the parameters of the kinetics of the reaction between OPA and AGM were determined as follows: when the initial concentration of AGM (5 × 10−6 M) is equal to that of the OPA, the results show that the reaction follows an overall 1.5 order kinetics with an initial rate of 5.1 × 10−7Mmin−1 and a half-life of 8.12 min. The partial order found in relation to the AGM is 0.8. This work shows that the excess of OPA accelerates the formation reaction of the complex.

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