The document discusses enzyme kinetics models including the Michaelis-Menten model and Lineweaver-Burk double reciprocal plot. The Michaelis-Menten model relates reaction rate to substrate concentration using kinetic constants Km and Vmax. It describes the enzyme-substrate reaction mechanism. The Lineweaver-Burk plot is a graphical representation that transforms the Michaelis-Menten equation into a straight line to determine Km and Vmax. It can distinguish competitive and noncompetitive enzyme inhibition patterns.
This presentation is about the kinetics of enzyme action , the Michaelis- Menten Model and kinetics of allosteric enzyme action in a simplified language.
This presentation deals with basics of enzyme kinetics and introduction to various plots which aid in understanding the mechanism of inhibition of enzymes.
This presentation is about the kinetics of enzyme action , the Michaelis- Menten Model and kinetics of allosteric enzyme action in a simplified language.
This presentation deals with basics of enzyme kinetics and introduction to various plots which aid in understanding the mechanism of inhibition of enzymes.
Some of the enzyme possess additional sites, known as allosteric sites besides the active site . Such as know as allosteric enzyme. The allosteric sites are unique place on the enzyme molecules allosteric enzyme have one or more allosteric site.
HISTRY
The term allosteric has been introduced by the two Noble Laureates JACOB AND MONOD to denote an enzyme site different from the active site which non competitively bands molecule other than the substrate and may influence the enzyme activity.
Properties of allosteric enzyme
Effector may be positive or negative, this effector regulate the enzyme activity . The enzyme activity is increased when a positive allosteric effector binds at the allosteric site known as activator site. On the other hand negative allosteric effector bind at the allosteric site called inhibitor site and inhibit the enzyme activity
Biological oxidation (part - III) Oxidative PhosphorylationAshok Katta
Biological oxidation (part - III) Oxidative Phosphorylation
- Mechanism of Oxidative Phosphorylation
-- Chemiosmotic theory
-P:O Ratio
Substrate Level Phosphorylation
Shuttle Systems for Oxidation of Extramitochondrial NADH
Active sites of the enzyme is that point where substrate molecule bind for the chemical reaction. It is generally found on the surface of enzyme and in some enzyme it is a “Pit” like structure
The active site is a three-dimensional cleft formed by groups that come from different parts of the amino acid sequence
The active site takes up a relatively small part of the total volume of an enzyme
Active sites are clefts or crevices
Substrates are bound to enzymes by multiple weak attractions.
The specificity of binding depends on the precisely defined arrangement of atoms in an active site.
it is bypass cycle of citric acid cycle.
it give the brief description of glyoxylate cycle.
it is the summary of glyoxylate cycle for m.sc, bsc, science students.
it is very important topic for entrance exam of biology stream.
Nucleotide Biosynthesis involves 2 processes. one is Denovo synthesis and other is Salvage pathway. An outline of both the processes has given in this presentation.
The flux of metabolites through metabolic pathways involves
catalysis by numerous enzymes. Active control of homeostasis is achieved by the regulation of only a small number of enzymes.
Some of the enzyme possess additional sites, known as allosteric sites besides the active site . Such as know as allosteric enzyme. The allosteric sites are unique place on the enzyme molecules allosteric enzyme have one or more allosteric site.
HISTRY
The term allosteric has been introduced by the two Noble Laureates JACOB AND MONOD to denote an enzyme site different from the active site which non competitively bands molecule other than the substrate and may influence the enzyme activity.
Properties of allosteric enzyme
Effector may be positive or negative, this effector regulate the enzyme activity . The enzyme activity is increased when a positive allosteric effector binds at the allosteric site known as activator site. On the other hand negative allosteric effector bind at the allosteric site called inhibitor site and inhibit the enzyme activity
Biological oxidation (part - III) Oxidative PhosphorylationAshok Katta
Biological oxidation (part - III) Oxidative Phosphorylation
- Mechanism of Oxidative Phosphorylation
-- Chemiosmotic theory
-P:O Ratio
Substrate Level Phosphorylation
Shuttle Systems for Oxidation of Extramitochondrial NADH
Active sites of the enzyme is that point where substrate molecule bind for the chemical reaction. It is generally found on the surface of enzyme and in some enzyme it is a “Pit” like structure
The active site is a three-dimensional cleft formed by groups that come from different parts of the amino acid sequence
The active site takes up a relatively small part of the total volume of an enzyme
Active sites are clefts or crevices
Substrates are bound to enzymes by multiple weak attractions.
The specificity of binding depends on the precisely defined arrangement of atoms in an active site.
it is bypass cycle of citric acid cycle.
it give the brief description of glyoxylate cycle.
it is the summary of glyoxylate cycle for m.sc, bsc, science students.
it is very important topic for entrance exam of biology stream.
Nucleotide Biosynthesis involves 2 processes. one is Denovo synthesis and other is Salvage pathway. An outline of both the processes has given in this presentation.
The flux of metabolites through metabolic pathways involves
catalysis by numerous enzymes. Active control of homeostasis is achieved by the regulation of only a small number of enzymes.
enzyme kinetics and michael menten’s constantManisha371125
this will give you a brief idea of how the enzyme works, how enzyme kinetics work, Michaelis constant (Km)
, Michaelis-Menten’s equation derivation, rate of reaction, Significance of Michaelis-Menten, Constant Km,
enzyme efficiently etc
What is enzyme?
How enzyme catalyze the reaction
Enzyme kinetics
History
Enzyme kinetic equation
Michaelis-menten equation
Michaelis-menten curve
Michaelis-menten equation derivation
Reversible inhibition
Two substrate reaction
Conclusion
References
A diffusion-limited enzyme catalyses a reaction so efficiently that the rate limiting step is that of substrate diffusion into the active site, or product diffusion out. This is also known as kinetic perfection or catalytic perfection. Since the rate of catalysis of such enzymes is set by the diffusion-controlled reaction, it therefore represents an intrinsic, physical constraint on evolution (a maximum peak height in the fitness landscape). Diffusion limited perfect enzymes are very rare. Most enzymes catalyse their reactions to a rate that is 1,000-10,000 times slower than this limit. This is due to both the chemical limitations of difficult reactions, and the evolutionary limitations that such high reaction rates do not confer any extra fitness.
Here you can find out the information about a very important topic of Biochemistry i.e. Enzyme Kinetics.
I have elaborated how enzymes kinetics works . The most important equation Michaelis-menten equation, and Burk plot.
#enzymes #biochemistry
Similar to Enzyme kinetics- michaelis menten model, lineweaver burk plot (20)
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
for beginners, providing thorough training in areas such as SEO, digital communication marketing, and PPC training in Noida. After finishing the program, students receive the certifications recognised by top different universitie, setting a strong foundation for a successful career in digital marketing.
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Introduction to AI for Nonprofits with Tapp NetworkTechSoup
Dive into the world of AI! Experts Jon Hill and Tareq Monaur will guide you through AI's role in enhancing nonprofit websites and basic marketing strategies, making it easy to understand and apply.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
3. INTRODUCTION
● Enzyme kinetics is the study of the rates of enzyme-catalysed chemical reactions. In
enzyme kinetics, the reaction rate is measured and the effects of varying the conditions
of the reaction are investigated. Studying an enzyme's kinetics in this way can reveal
the catalytic mechanism of this enzyme, its role in metabolism, how its activity is
controlled, and how a drug or a modifier (inhibitor or activator) might affect the rate.
● Enzymes are natural proteins which act as catalysts that speed up the rate of specific
chemical reactions. They either help create or break down molecules.
● An enzyme (E) is typically a protein molecule that promotes a reaction of another
molecule, its substrate (S).
4. ● This binds to the active site of the enzyme to produce an enzyme-substrate complex ES,
and is transformed into an enzyme-product complex EP and from there to product P, via
a transition state. The series of steps is known as the mechanism.
● There are many practical uses of enzyme kinetics. For example, the kinetic constants
can help explain how enzymes work and assist in the prediction of the behavior of
enzymes in living organisms.
5. MICHAELIS AND MENTEN
● In biochemistry, Michaelis–Menten kinetics is one of the best-known models of
enzyme kinetics. It is named after German biochemist Leonor Michaelis and Canadian
physician Maud Menten.
● The model takes the form of an equation describing the rate of enzymatic reactions, by
relating reaction rate V i.e., Enzyme velocity (rate of formation of product [P] to [S] )
and the concentration of a substrate S.
6. MICHAELIS
● Leonor Michaelis was a German biochemist, physical chemist, and physician, known
for his work with Maud Menten on enzyme kinetics in 1913, as well as for work on
enzyme inhibition, pH and quinones.
MENTEN
● Maud Leonora Menten was a Canadian bio-medical and medical researcher who made
significant contributions to enzyme kinetics and histochemistry. She is primarily known
for her work with Leonor Michaelis on enzyme kinetics in 1913.
7. ENZYME VELOCITY
➢ Enzyme Velocity is the rate of an enzyme-catalyzed reaction is often called its velocity.
Enzyme velocities are normally reported as values at time zero (initial velocity, symbol
Vo; μmol min-1). This is because the rate is fastest at the point where no product is yet
present as the substrate concentration is greatest before any substrate has been
transformed to product.
SUBSTRATE CONCENTRATION
➢ Substrate concentration is the amount of substrate present that can be turned into
product and is most commonly measured in molarity (moles per liter). The
concentration of substrates is used to measure enzyme activity
8. MICHAELIS MENTEN MODEL
● This model takes the form of an equation describing the rate of enzymatic reactions, by
relating reaction rate v (rate of formation of product [P]) to the concentration of a
substrate [S].
V0 = Vmax [S]
Km + [S]
● The Michaelis–Menten model uses the following concept of enzyme catalysis:
9. ● The enzyme (E), combines with its substrate (S) to form an enzyme–substrate complex
(ES). The ES complex can dissociate again to form E + S, or can proceed chemically to
form E and the product P.
● The rate constants k1, k2 and k3 describe the rates associated with each step of the
catalytic process.
● It is assumed that there is no significant rate for the backward reaction of enzyme and
product (E + P) being converted to ES complex. [ES] remains approximately constant
until nearly all the substrate is used, hence the rate of synthesis of ES equals its rate of
consumption over most of the course of the reaction; that is, [ES] maintains a steady
state.
● From this concept, the Michaelis–Menten equation was derived.
10. V0 = Vmax [S] = Michaelis menten equation
Km + [S]
● The normal pattern of dependence of enzyme rate on substrate concentration ([S]) is
that at low substrate concentrations a doubling of [S] will lead to a doubling of the
initial velocity (V0). However, at higher substrate concentrations the enzyme becomes
saturated and further increases in [S] lead to very small changes in V0.
● This occurs because at saturating substrate concentrations, all of the enzyme molecules
have bound substrates. The overall enzyme rate is now dependent on the rate at which
the product can dissociate from the enzyme, and adding further substrate will not affect
this. In situations where the substrate concentration is saturating, a doubling of the
enzyme concentration will lead to a doubling of V0.
12. ● The following equation, known as the Equation of enzyme machanism, is used to
describe the kinetics of enzymes:
● Where E, S, ES, and P represent enzyme, substrate, enzyme–substrate complex, and
product, respectively.
● The Kf, Kr, & Kcat denote “the rate constants for the "forward" binding” and "reverse
unbinding of substrate”, and for the “catalytic conversion of substrate into product”,
respectively.
K1 = Kr, K2 = Kf, K3 = Kcat
13. ● According to Michaelis menten equation. The first linear part shown in the graph is
known as the 1st Order Kinetics. The linear increase in Velocity with increase in
Substrate concentration.
● The graph shows a plateau region where increase in substrate concentration no longer
increases with the velocity of the reaction. At this stage velocity has reached the
maximum velocity. Which is denoted as Vmax. This plateau region is called as 0th
order kinetics which means Velocity is independent of substrate concentration.
● The Michaelis Menten equation explains this curve (plateau) mathematically. The aim
of this equation is to find the mathematical relation between V0, Vmax, and Km. Such
that, both 0 Order and 1st Order kinetics can be explained.
14. ● According to Laws of mass action,
Kf [E][S] = Kr [ES]
● If we take the ratio of this equation we will get,
[E][S] = Kr = Kd
[ES] Kf
● This Kd is called the Dissociation constant. According to the Pseudo - steady - state
hypothesis, The state of ES in the first equation remains constant.
[ES] formation = [E][S] breakdown
So,
ES formation = Kf [E][S]
ES breakdown = Kr [ES] + Kcat[ES]
15. Ie we say that, Kf [E][S] = Kr [ES] + Kcat[ES]
Taking [ES] common gives, Kf [E][S] = [ES] (Kr+Kcat)
Taking the ratio, [E][S] = Kr+Kcat = Km (Michaelis menten constant)
[ES] Kf
According to our aim, our aim is to find the mathematical relation between V0, Vmax, and
Km. We have,
[E][S] = Km.
[ES]
Velocity is equal to rate of production per unit time. Ie, V0 = d[P]
dt
Velocity Vo depends on the breakdown of the ES complex. Ie,
V0 = Kcat [ES]
16. ● There are some molecules which are not bound with the substrate and other enzyme
molecules bound with the substrate, so that total enzyme concentration can be given as
E0 = E + ES
● When all enzyme molecules are bound with the substrate there is no free enzyme left,
hence, E0 = E+ES where E=0, E0=ES. As all enzymes are occupied by the substrates
the velocity reaches maximum velocity or Vmax.
● Therefore Vmax = Kcat[E0] were E0 = ES. Now let's rearrange the equation to get one
single equation.
E0 = E+ES
Taking E on the other side, (E0 - ES) = E or E = (E0 - ES)
This E can be replaced in the equation of Km.
Km = [E][S] = (E0 - ES)[S]
[ES] [ES]
17. Now let's multiply [S] with the term E = E0 - ES, So we get
Km = (E0 - ES)[S] = [E0][S] - [ES][S] = [E0][S] - [S]
[ES] [ES] [ES] [ES] 1
Rearranging the equation,
Km = [E0][S] - [S]
[ES] 1
Now the term [E0] can be replaced by Vmax
Kcat
Km = Vmax[S] - [S]
Kcat [ES] 1
The product of Kcat [ES] = V0
Km = Vmax[S] - [S]
V0 1
18. If we take -[S] on the other side with Km, then we get,
Km + [S] = Vmax [S]
V0
Rearranging this equation finally we get,
This Equation is called the Michaelis menten equation. Which says when substrate
concentration is very large, The value of Km will be very less, as compared to the value of
S. Hence, Km can be ignored when compared to S.
Then equation becomes
V0 = Vmax [S]
[S]
[S] can be cancelled. Finally we will get
V0 = Vmax.
V0 = Vmax [S]
Km + [S]
19. The Michaelis Menten constant in turn is defined as follows:
● The Michaelis menten constant is equal to the substrate concentration at which the
enzyme converts substrates into products at half its maximal rate and hence is related to
the affinity of the substrate for the enzyme.
● The catalytic constant Kcat is the rate of product formation when the enzyme is
saturated with substrate and therefore reflects the enzyme's maximum rate. The rate of
product formation is dependent on both how well the enzyme binds substrate and how
fast the enzyme converts substrate into product once substrate is bound. For a
kinetically perfect enzyme, every encounter between enzyme and substrate leads to
product and hence the reaction velocity is only limited by the rate the enzyme
encounters substrate in solution. Hence the upper limit for Kcat/ Km is equal to the rate
of substrate diffusion.
20. Ie, Simply., V0 = Vmax
2
Applying in the main equation
V0 = Vmax = Vmax [S] = 1 = [S] _
2 Km + [S] 2 Km + [S]
Removing Vmax on both sides, and rearranging the equation
1 = 2 [S] _
Km + [S]
Km = 2 [S] - [S]
Km = [S] (definition of michaelis menten constant)
21. APPLICATIONS
➢ The constant Km is a measure of how efficiently an enzyme
converts a substrate into a product.
➢ The equation can also be used to describe the relationship
between ion channel conductivity and ligand concentration.
22. LINEWEAVER AND BURK - INTRODUCTION
● In biochemistry, the Lineweaver–Burk plot (or double reciprocal plot) is a graphical
representation of the Lineweaver–Burk equation of enzyme kinetics, described by Hans
Lineweaver and Dean Burk in 1934.
● The Lineweaver–Burk plot for inhibited enzymes can be compared to non inhibited
enzymes to determine how the inhibitor is competing with the enzyme.
23. LINEWEAVER & BURK
● Hans Lineweaver was an American physical chemist, who is credited (misleadingly)
with introducing the double-reciprocal plot or Lineweaver–Burk plot.
● The paper containing the equation was co-authored by Dr. Dean Burk,who was an
American biochemist, medical researcher, and a cancer researcher at the Kaiser
Wilhelm Institute and the National Cancer Institute, and was entitled "The
Determination of Enzyme Dissociation Constants (1934)".
24. LINEWEAVER-BURK DOUBLE RECIPROCAL PLOT
● Since, Vmax is achieved at infinite substrate concentration, it is impossible to estimate
Vmax and hence Km from a hyperbolic plot. Because of this difficulty, the
Michaelis–Menten equation was transformed into an equation for a straight line by
Lineweaver and Burk.
● The Lineweaver–Burk plot (or double reciprocal plot) is a graphical representation of
the Lineweaver–Burk equation of enzyme kinetics. This plot is a derivation of the
Michaelis–Menten equation and is represented as
Where,
● V is reaction velocity ( the reaction rate)
● Km is the Michaelis menten constant
● Vmax is the maximum reaction velocity, and
● S is the substrate concentration
25. LINEWEAVER BURK EQUATION
The plot provides a useful graphical method for analysis of the Michaelis–Menten
equation, as it is difficult to determine precisely the Vmax of an enzyme-catalysed reaction:
26. We have the Michaelis menten equation
V0 = Vmax[S] =
Km + [S]
Taking the reciprocal gives
_1 = Km + [S]
V0 Vmax[S]
_1 = Km + [S]__
V0 Vmax[S] Vmax[S]
_1 = Km + 1__
V0 Vmax[S] Vmax
27. _1 = Km + 1__
V0 Vmax[S] Vmax
If we compare this equation with equation of straight line Y = mx + c
Y = 1 _
V0
M = Km
Vmax
X = 1 _
[S]
C = 1 _
Vmax
28. ● Reversible enzyme inhibitors can be classified as either competitive or noncompetitive, and
can be distinguished via a Lineweaver–Burk plot. It is a useful way of determining how an
inhibitor binds to an enzyme.
● Competitive inhibition can be recognized by using a Lineweaver–Burk plot if V0 is
measured at different substrate concentrations in the presence of a fixed concentration of
inhibitor. A competitive inhibitor increases the slope of the line on the Lineweaver–Burk
plot, and alters the intercept on the x-axis (since Km is increased), but leaves the intercept on
the y- axis unchanged (since Vmax remains constant).
● Noncompetitive inhibition can also be recognized on a Lineweaver–Burk plot since it
increases the slope of the experimental line, and alters the intercept on the y-axis (since
Vmax is decreased), but leaves the intercept on the x-axis unchanged (since Km remains
constant).
29. APPLICATIONS
● It is used to determine important terms in enzyme kinetics, such as Km and Vmax,
before the wide availability of powerful computers and non-linear regression software.
● This plot gives a quick, visual impression of the different forms of enzyme inhibition.
30. CONCLUSION
The two main principles in Enzyme kinetics are Michaelis menten kinetics and Lineweaver burk
plot and we also discussed two constants in Enzyme kinetics ie: Km and Vmax. Michaelis–Menten
kinetics is one of the best-known models of enzyme kinetics. It is named after German biochemist
Leonor Michaelis and Canadian physician Maud Menten. The model takes the form of an equation
describing the rate of enzymatic reactions, by relating reaction rate V ( rate of formation of product
[P] to [S] ) and the concentration of a substrate S. According to the Lineweaver burk double
reciprocal plot, It is a graphical representation of the Lineweaver–Burk equation of enzyme kinetics,
described by Hans Lineweaver and Dean Burk in 1934. The Lineweaver–Burk plot for inhibited
enzymes can be compared to no inhibitor to determine how the inhibitor is competing with the
enzyme. In this plot Michaelis –Menten equation was transformed into an equation for a straight
line by Lineweaver and Burk.
31. REFERENCES
● David Hames and Nigel Hooper (2005). Biochemistry. Third ed. Taylor & Francis
Group: New York.
● Dean Burk/wikipedia
● Enzyme kinetics/Wikipedia
● Hans Lineweaver/wikipedia
● Leonor Michaelis/wikipedia
● Maud Leonora Menten/wikipedia
● Michaelis menten model/microbenotes
● Michaelis-Menten Model/wikipedia.
● Srinivasan, Bharath (2021-07-16). "A Guide to the Michaelis‐Menten equation: Steady
state and beyond"
● Stryer L, Berg JM, Tymoczko JL (2002). "Section 8.4: The Michaelis-Menten Model
Accounts for the Kinetic Properties of Many Enzymes". Biochemistry (5th ed.). San
Francisco: W.H. Freeman. ISBN 0-7167-4955-6.)