Name: graphing enzyme kinetics data
Uploaded: 2023-04-30T10:09:42.000Z
Duration: 1 min 19 s
Channel: JoVE

K12 Standards

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North Carolina K-12 Science Standard Course of Study

9-12

Biology

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms- Structures and Processes

LS.Bio.1.2

Introduction to Enzyme Kinetics

Die Enzymkinetik untersucht die Raten enzymkatalysierter Reaktionen. Die Häufigkeit wiederholter Experimente bei unterschiedlichen Substratkonzentrationen wird überwacht, indem die Konzentration des verbrauchten Substrats oder des im Laufe der Zeit gebildeten Produkts gemessen wird.
Diese Ergebnisse können grafisch dargestellt werden, um zu zeigen, wie sich die Substratkonzentration auf die Geschwindigkeit oder Geschwindigkeit einer Reaktion auswirkt.
Die Reaktionsgeschwindigkeit nimmt mit zunehmender Substratmenge bei niedrigen Konzentrationen linear zu, beginnt aber bei höheren Konzentrationen ein Plateau zu erreichen. Die Geschwindigkeit nähert sich einer maximalen Geschwindigkeit oder Vmax – der Rate, bei der das Enzym vollständig mit dem Substrat gesättigt ist.
Die Enzymaffinität misst, wie stark oder schwach ein Enzym an sein Substrat bindet und wird durch KM, die Michaelis-Konstante, quantifiziert. Der Wert von KM ist gleich der Substratkonzentration, wenn die Rate 50 % des V max beträgt.
Ein kleines KM zeigt an, dass ein Enzym eine hohe Substrataffinität hat und umgekehrt. Ein Enzym mit einem größeren KM benötigt im Vergleich zu einem Enzym mit einem niedrigeren KM höhere Substratkonzentrationen, um sich seiner maximalen Geschwindigkeit zu nähern.

<ol>
	<li>Voet D. und Voet J.G. Biochemie, 4. Aufl. John Wiley & Sons, 2010 pp-487-492</li>
	<li>Berg J.M., Tymoczko J.L. und Stryer L. Biochemie, 5. Aufl., W.H. Freeman and Co., Abschnitt 8.4</li>
</ol>

introduction to enzyme kinetics

Enzyme kinetics studies the rates of biochemical reactions. Scientists monitor the reaction rates for a particular enzymatic reaction at various substrate concentrations. Additional trials with inhibitors or other molecules that affect the reaction rate may also be performed.

The experimenter can then plot the initial reaction rate or velocity (Vo) of a given trial against the substrate concentration ([S]) to obtain a graph of the reaction properties. For many enzymatic reactions involving a single substrate, this data fits the Michaelis-Menten equation, an equation derived by Leonor Michaelis and Maud Menten.

<img alt="Eq1" src="/files/ftp_upload/12164/12164_Equation 1.png" style="text-align: center; height: 70px;" />

The equation estimates the maximum velocity (Vmax) and the Michaelis constant (KM) for the enzyme being studied and is based on the following assumptions:

<ol>
	<li>No product is present at the start of the reaction.</li>
	<li>The rate of enzyme-substrate complex formation equals the rate of dissociation and breakdown into products.</li>
	<li>The enzyme concentration is minimal compared to the substrate concentration.</li>
	<li>Only the initial reaction rates are measured.</li>
	<li>The enzyme is present either in the free form or in the enzyme-substrate complex.</li>
</ol>

Different rearrangements of the Michaelis-Menten equation, such as the Lineweaver-Burke, Eadie-Hofsteot, and Hanes-Woolf plots, are alternate ways to graph kinetic parameters. The Lineweaver-Burke or double reciprocal plot is often used to estimate the KM and the Vmax. The plot uses the reciprocals values of the x and y-axis from the Michaelis-Menten plot. Mathematically, the y-intercept equals 1/Vmax, and the x-intercept equals −1/KM.

The Lineweaver-Burke plot can be used to visually differentiate between inhibitor types – competitive, non-competitive, and uncompetitive. Different rearrangements of the Michaelis-Menten equation, such as the Eadie-Hofstee and Hanes-Woolf plots, are also used to determine kinetic parameters.

Die Enzymkinetik untersucht die Geschwindigkeit biochemischer Reaktionen. Wissenschaftler überwachen die Reaktionsgeschwindigkeiten für eine bestimmte enzymatische Reaktion an verschiedenen Substraten ...

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Cell Biology

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KEY TERMS AND CONCEPTS

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graphing enzyme kinetics data

Enzyme kinetics uses graphs to show how fast a biochemical reaction happens. Scientists measure the initial reaction rate, or velocity (Vo), at different substrate concentrations ([S]). They may also run trials with inhibitors or other molecules that change the reaction rate.

The results are often plotted as Vo against [S]. For many single-substrate reactions, the data fits the Michaelis-Menten equation. This equation, named after Leonor Michaelis and Maud Menten, helps estimate the maximum velocity (Vmax) and the Michaelis constant (KM) for the enzyme being studied.

The Michaelis-Menten model is based on a few assumptions. No product is present at the start of the reaction. The rate of enzyme-substrate complex formation equals the rate of its breakdown into products and its dissociation. The enzyme concentration is minimal compared with the substrate concentration. The model also uses only the initial reaction rates. In this setup, the enzyme is either free or bound in an enzyme-substrate complex.

Scientists can also use other graph forms to analyze the same data. The Lineweaver-Burke plot, also called the double reciprocal plot, uses the reciprocal values from the Michaelis-Menten graph. On this plot, the y-intercept equals 1/Vmax and the x-intercept equals -1/KM. It is often used to estimate KM and Vmax and to visually distinguish competitive, non-competitive, and uncompetitive inhibitors.

Other rearrangements of the Michaelis-Menten equation include the Eadie-Hofstee plot and the Hanes-Woolf plot. These graphs are also used to determine kinetic parameters. Together, these methods help scientists compare reaction behavior and understand how enzymes respond to substrate concentration and inhibitors.

Graphing Enzyme Kinetics Data

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Enzyme kinetics uses graphs to show how fast a biochemical reaction happens. Scientists measure the initial reaction rate, or velocity (Vo), at different substrate concentrations ([S]). They may also run trials with inhibitors or other molecules that change the reaction rate. The results are often plotted as Vo against [S]. For many single-substrate reactions, the data fits the Michaelis-Menten equation. This equation, named after Leonor Michaelis and Maud Menten, helps estimate the maximum...

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Learn how enzyme kinetics data is graphed with Michaelis-Menten, Lineweaver-Burke, Eadie-Hofstee, and Hanes-Woolf plots to estimate Vmax and KM.

Graphing Enzyme Kinetics Data | JoVE K12

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HIgh School

BIO.2

BIO.2.c

Chemistry

CH.7

CH.7.f

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Life Science

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SC.912.L.18.4

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Graphing Enzyme Kinetics Data

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