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Enzyme kinetics,

regulation
and
catalysis
Content
Active site
Coenzyme,
Nomenclature
Catalysis
Kinetics
Inhibition
Regulation
Special class
Introduction

Biocatalyst—homeostasis

External—Blood clotting

Internal—metabolism

Plasma membrane—acts to signal


Active site of enzyme
Coenzyme and cofactors
Cofactor

Organic Inorganic

Coenzyme Metal ion

Fe-S cluster
Prosthetic
group
Enzyme nomenclature

 Oxidoreductase
 Transferase
 Hydrolase
 Lyase
 Isomerase
 Ligase
Enzyme kinetics

 Equation

 Significance

 Plots
Mechanism of action
Enzyme velocity vs substrate
concentration
Michalis - Menten equation

Vmax [S]
Velosity = V =
[S] + KM
Significance of Km

 Km is [S]; when V 0 = Vmax/2

 kM= k - 1 +k + 2 /k + 1

 Km= Ks= k - 1 /k + 1 =[E][S]/[ES];

When k - 1 >>>k + 2

 k c a t / K m =K e f f
Assumptions

 The production of product


is linear

 The concentration of
substrate greater than
enzyme

 A single enzyme forms


the product
Assumptions
 There is negligible

product without enzyme

 No cooperativity

 Neither substrate nor

product -allosteric

modulator
Plots
Enzyme Inhibition

Enzyme Inhibition

Irreversible Reversible
Competitive

Un-competitive

Non-competitive
Irreversible inhibition
Reversible Competitive inhibition
Reversible
Uncompetitive inhibition
Reversible
Non competitive inhibition
Enzyme regulation

 Allosteric Control

 Covalent Modification

 Protein Processing
Allosteric control
Aspartate trans carbamoylase
Catalytic
trimer

Regulatory
Dimer
Regulatory
Dimer

Regulatory
Dimer

Catalytic
trimer
Cooperativity
Feedback Inhibition
Covalent modification of enzyme
Protein processing Zymogen
activation
Enzyme catalysis

 Bond strain

 Proximity/Orientation effect

 Electrostatic catalysis

 Covalent catalysis

 Quantum tunnelling
Bond strain
Proximity/orientation
effect
Electrostatic catalysis

 Acid base catalysis

 Metal ion catalysis


Acid-Base catalysis
Metal ion catalysis
Covalent catalysis

 Ser (OH) class—acyl enzyme,

phosphoenzyme

 Cys (SH) class---Gly-3P-DH

 His(Im) class-succinyl CoA

synthase

 Lys(NH2) class—Fru-

diphosphate-aldolase
Quantum tunnelling
Ribozyme
Abzyme
Isozyme and Allozyme
Bi-substrate reaction

 Sequential reaction--Ordered

sequential and Random

sequential

 Ping-Pong reaction
Emil Fischer
(1852-1919)
Lock and key model
D. Koshland
(1920-2007)
Leonor Michaelis Maud Menten
(1875–1949) (1879–1960)
Linear equation for
Lineweaver-Burk plot
Succinate Dehydrogenase
-competitive inhibitor
Reaction
Positive and negative
co-operativity