Correcting Lineweaver—Burt calculations of V and Km

Glick, Ned; Landman, Amiram D.; Roufogalis, Basil D.

doi:10.1016/0968-0004(79)90182-8

Cited by 13 publications

(5 citation statements)

References 7 publications

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“…They generally agreed within 10%, and the mean was used. However, when a significant amount of substrate was consumed (≥15% at initial concentrations below K m ), the rate from each time interval was handled separately and plotted against the average substrate concentration in the time interval, as recommended by Glick et al . (1979).…”

Section: Methodsmentioning

confidence: 99%

Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form

Londesborough¹,

Vuorio²

1991

Microbiology

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Journal of General Microbio/ogyA protein of about 800 kDa with trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP) activity was purified from bakers' yeast. This TPS/P complex contained 57,86 and 93 kDa polypeptides. The 86 and 93 kDa polypeptides both appeared to be derived from a polypeptide of at least 115 kDa in the native enzyme. A TPS-activator (a dimer of 58 kDa subunits) was also purified. It decreased the Michaelis constants for both UDP-glucose (three-fold) and glucose 6-phosphate (G6P) (4-5-fold), and increased TPS activity at 5 mM-UDP-glucose/lO ~M -G~P about three-fold. It did not affect TPP activity. The purification of TPS/P included an endogenous proteolytic step that increased TPS activity about three-fold and abolished its requirement for TPSactivator, but did not change TPP activity. This activation was accompanied by a decrease of some 20 kDa in the molecular mass of a cluster of SDS-PAGE bands at about 115 kDa recognized by antiserum to pure TPS/P, but by no change in the 57 kDa band. Phosphate inhibited TPS activity (Ki about 5 mM), but increased TPP activity about six-fold (K, about 4 mM). Phosphate (6 mM) stimulated the synthesis of trehalose from G6P and UDPglucose and decreased the accumulation of trehalose 6-phosphate.

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Section: Methodsmentioning

confidence: 99%

Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form

Londesborough¹,

Vuorio²

1991

Microbiology

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“…Based on a comparison of the number of citations, Glick, Landman, and Roufogalis (1979) conclude that this analysis procedure still appears to be widely used by enzyme researchers, despite the fact that the simulation analyses by Dowd and Riggs (1965) and Colquhoun (1969) showed that this procedure is by far the worst of all proposed estimation methods. The oldest and still most widely used procedure is based on equation (2), which is known as the Lineweaver-Burk plot.…”

Section: Introductionmentioning

confidence: 96%

Statistical Analysis of the Michaelis-Menten Equation

Raaijmakers¹

1987

Biometrics

158

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An application of the method of maximum likelihood (ML) is described for analysing the results of enzyme kinetic experiments in which the Michaelis-Menten equation is obeyed. Accurate approximate solutions to the ML equations for the parameter estimates are presented for the case in which the experimental errors are of constant relative magnitude. Formulae are derived that approximate the standard errors of these estimates. The estimators are shown to be asymptotically unbiased and the standard errors observed in simulated data rapidly approach the theoretical lower bound as the sample size increases. The results of a large-scale Monte Carlo simulation study indicate that for data with a constant coefficient of variation, the present method is superior to other published methods, including the conventional transformations to linearity and the nonparametric technique proposed by Eisenthal and Cornish-Bowden (1974, Biochemical Journal 139, 715-720). Finally, the present results are extended to the analysis of simple receptor binding experiments using the general approach described by Munson and Rodbard (1980, Analytical Biochemistry 107, 220-239).

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“…There is a certain difference between Vm and Km calculated by the Lineweaver buck method and the Wilkinson statistical method. The Wilkinson statistical method is considered to be reliable [ 26 ]. Therefore, the Michaelis equation obtained by the Wilkinson statistical method is finally selected as the final result.…”

Section: Resultsmentioning

confidence: 99%

Kinetics, Thermodynamics and Mechanism of Enzymatic Degradation of Zearalenone in Degummed Corn Oil

Zhao

Xie

Jin

et al. 2022

Toxins

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The kinetics and thermodynamics of the enzymatic degradation of zearalenone (ZEN) in degummed corn oil were investigated by analyzing the impacts of temperature, pH, ZEN hydrolase dosage and ZEN concentration on the initial reaction rate. The kinetic study found that the maximum reaction rate was 0.97 μmol × kg−1 min−1, the Michaelis constant (Km) was 11,476 μmol × kg−1 and the Michaelis equation was V = 0.97[S]/(11,476 + [S]). The thermodynamic study showed that the activation energy (Ea) was 70.37 kJ·mol−1, the activation enthalpy change of the reaction (ΔH) > 0, the free energy of activation (ΔG) > 0 and the activation entropy change (ΔS) < 0, indicating the reaction could not be spontaneous. The reaction mechanism of ZEN was studied by a hybrid quadrupole orbitrap mass spectrometer. It was found that ZEN first generated the intermediate G/L/D/W-ZEN+H2O, followed by generating the intermediate W-ZEN-H2O under the action of a degrading enzyme. Then, the lactone bond was opened to produce C18H24O6, and finally the decarboxylation product C17H24O4 formed automatically.

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Correcting Lineweaver—Burt calculations of V and Km

Cited by 13 publications

References 7 publications

Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form

Trehalose-6-phosphate synthase/phosphatase complex from bakers’ yeast: purification of a proteolytically activated form

Statistical Analysis of the Michaelis-Menten Equation

Kinetics, Thermodynamics and Mechanism of Enzymatic Degradation of Zearalenone in Degummed Corn Oil

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