CHE 505 REACTION ENGINEERING II

TOPIC 4
BIOCATALYST
REACTION
BY: MIRADATUL NAJWA MUHD RODHI
PART 2
MECHANISTIC MODELS FOR
SIMPLE ENZYME KINETICS
 Enzyme Kinetics
Enzyme kinetics refers to the quantitative analysis of all factors that
determine the catalytic potential of an enzyme

Enzyme activity represents the maximum catalytic potential of an

enzyme that is reflected by the initial rate of the catalyzed reaction.

Several factors affect the expression of such potential, being the most
important the concentrations of active enzyme, substrates and
inhibitors, temperature and pH

Reaction rates in enzyme kinetics refer always to initial reaction rates

where the maximum catalytic potential of the enzyme is expressed and
many factors affecting it (i.e. substrate depletion, accumulation of
inhibitory products, enzyme inactivation, reverse reaction) are irrelevant.
Source from: Gardossi , L. et al (2010). GUIDELINES FOR REPORTING OF BIOCATALYTIC REACTIONS, Trends in Biotechnology
 Chemical Kinetics
• Rate: measure product
formed per second
• Rate slows as reactant
disappears
• Measure initial rate
 Enzyme Kinetics
Homogeneous Enzyme Kinetics
→catalysts occupy the same phase as the
reaction mixture

Heterogeneous Enzyme Kinetics

→catalysts occupy a different phase

Homogeneous catalysts allow for greater mixing and interaction with the reaction mixture
than heterogeneous catalysts.
 Simple Mechanisms
• Chemical mechanism

• Enzyme Catalyzed
Major approaches used in developing a rate expression of enzyme catalyzed
reactions

Rapid-Equilibrium
Henri proposed the effect of substrate concentration on enzyme kinetics :
Conversion of substrate into product involved a reversible reaction between enzyme and substrate
to form an active intermediate that brakes down delivering the product.

Michaelis and Menten (1913) who proposed the first formal hypothesis for enzyme catalysis based
on two sequential steps, as suggested by Henri:
•In the first step the substrate is captured in the active site of the enzyme
•In the second step the amino acid residues at that site chemically process the substrate to
transform it into product, which is subsequently released to let the enzyme free and available for
the next catalytic round.

Equilibrium hypothesis was formulated based on the examination of hydrolytic reactions which in
aqueous milieu are virtually irreversible. However, for reversible reactions (S↔P) that hypothesis
is not applicable and steady-state equations must be derived.
Quasi-Steady-State

A few years later, Briggs and Haldane

(1925) argued against the validity of
the rapid equilibrium hypothesis and
proposed a steady-state hypothesis
according to which, after a very short
transient phase, the ES complex
remains constant throughout the
whole reaction period.
 Michaelis-Menten Kinetics
• Km is the [S] at which the reaction
reaches half its maximum velocity
Vmax [ S ]
• Physical meaning (assuming v
equilibrium binding): Km is the Km  S
dissociation constant for ES
• Km is [S] at which enzyme is half-
bound
• Km is measure of affinity of
enzyme for S
• Low Km is tight binding
• Low [S]
– Rate very dependent on [S]
– Binding is rate limiting

• High [S]
– Rate independent
What are two things contribute to the – Saturation of E
maximum velocity limit? – Chemistry is rate limiting

• Amount of enzyme
• Chemical ability of enzyme (catalytic
constant)
Deriving Michaelis-Menten Equation

Vmax [ S ]
v
Km  S