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

K12 Standards

north-carolina-k-12-science-standard-course-of-study

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

효소 반응속도론은 효소 촉매 반응의 속도를 연구합니다. 다양한 기질 농도에서 반복되는 실험의 속도는 시간이 지남에 따라 소비된 기질 또는 형성된 제품의 농도를 측정하여 모니터링됩니다.
이러한 결과는 기질 농도가 반응 속도 또는 속도에 어떻게 영향을 미치는지 보여주기 위해 그래프로 표시할 수 있습니다.
반응 속도는 낮은 농도에서 기질의 양이 증가함에 따라 선형적으로 증가하지만 더 높은 농도에서는 안정되기 시작합니다. 속도는 최대 속도 또는 Vmax에 접근합니다 – 효소가 기질로 완전히 포화되는 속도입니다.
효소 친화도는 효소가 기질에 얼마나 강하게 또는 약하게 결합하는지를 측정하고 Michaelis 상수인 KM으로 정량화합니다. KM의 가치는 비율이 Vmax의 50%일 때 기질 농도와 동등합니다.
작은 KM은 효소가 높은 기질 친화도를 갖는다는 것을 나타내며 그 반대의 경우도 마찬가지입니다. KM이 더 큰 효소는 KM이 더 낮은 효소에 비해 최대 속도에 접근하기 위해 더 높은 기질 농도가 필요합니다.

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

효소 동역학 소개

효소 동역학은 생화학 반응 속도를 연구합니다. 과학자들은 다양한 기질 농도에서 특정 효소 반응에 대한 반응 속도를 모니터링합니다. 반응 속도에 영향을 미치는 억제제나 기타 분자를 사용한 추가 시험도 수행될 수 있습니다.

그런 다음 실험자는 기질 농도([S])에 대해 주어진 시험의 초기 반응 속도 또는 속도(Vo)를 플롯하여 반응 특성의 그래프를 얻을 수 있습니다. 단일 기질과 관련된 많은 효소 반응의 경우 이 자료는 레오노르 미카엘리스와 모드 멘텐이 도출한 방정식인 미카엘리스-멘텐 방정식에 적합합니다.

<img alt="Eq1" src="/files/ftp_upload/12164/12164_Equation 1.png" style="text-align: center; height: 70px;" />

방정식은 연구 중인 효소의 최대 속도(Vmax)와 미카엘리스 상수(KM)를 추정하며 다음 가정을 기반으로 합니다:

<ol>
	<li>반응이 시작될 때 생성물이 존재하지 않습니다.</li>
	<li>효소-기질 복합체 형성 속도는 해리 및 생성물로의 분해 속도와 동일합니다.</li>
	<li>효소 농도는 기질 농도에 비해 최소입니다.</li>
	<li>초기 반응 속도만 측정됩니다.</li>
	<li>효소는 유리 형태 또는 효소-기질 복합체로 존재합니다.</li>
</ol>

라인위버-버크, 이디-호프스티 및 한스-울프 방정식과 같은 미카엘리스-멘텐 방정식의 다양한 재배열은 운동 매개변수를 그래프로 표시하는 대체 방법입니다. 라인위버-버크 또는 이중 역수 도표는 KM 및 Vmax를 추정하는 데 종종 사용됩니다. 이 플롯은 미카엘리스-멘텐 방정식의 x축과 y축의 역수 값을 사용합니다. 수학적으로 y절편은 1/Vmax이고 x절편은 −1/KM입니다.

라인위버-버크 방정식을 사용하면 억제제 유형(경쟁적, 비경쟁적, 비경쟁적)을 시각적으로 구분할 수 있습니다. 이디-호프스티 및 한스-울프 방정식과 같은 미카엘리스-멘텐 방정식의 다양한 재배열도 운동 매개변수를 결정하는 데 사용됩니다.

효소 반응 속도학은 생화학 반응의 속도를 연구합니다. 과학자들은 다양한 기질에서 특정 효소 반응에 대한 반응 속도를 모니터링합니다 ...

Education

교육

JoVE Core

Cell Biology

세포생물학

Unit 1

제1단원

Energy and Catalysis

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KEY TERMS AND CONCEPTS

주요 용어 및 개념

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

the first law of thermodynamics

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introduction to enzyme kinetics

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...

turnover number and catalytic efficiency

How Fast Enzymes Work in Digestion

Enzymes can work at very different speeds, and their turnover number shows how fast they convert substrate molecules into products. Turnover number is the maximum number of substrate molecules an enzyme can transform per unit time. For most enzymes, this value is between 1 and 1000 molecules per second. Some enzymes are especially fast. Catalase has the highest known turnover number and can convert up to 2.8×10^6 molecules of hydrogen peroxide into water and oxygen each second. Lysozyme has...

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Diffusion-Controlled Enzymes and Reaction Speed

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key terms and concepts

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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JoVE Core Cell Biology illustrates complex cellular processes in easy-to-understand yet comprehensive animated video lessons. This collection focuses on the essential components and functions of cells, including organelles and membranes, cellular transport, and cell energetics. It is complemented by scientist-in-action videos that demonstrate the application of cell biology in real-world research experiments.

Virginia  Standards of Learning

HIgh School

BIO.2

BIO.2.c

Chemistry

CH.7

CH.7.f

Florida Department of Education

High School

Life Science

SC.912.L.18

SC.912.L.18.4

SC.912.L.18.11

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Graphing Enzyme Kinetics Data

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