Chemical
Mechanism
An important goal of measuring
enzyme kinetics is to determine the
chemical mechanism of an enzyme
• Kinetic measurements taken under various solution conditions
or on slightly modified enzymes or substrates often shed light
on this chemical mechanism, as they reveal the rate-determining
step or intermediates in the reaction.
• Isotopes can also be used to reveal the fate of various parts of
the substrate molecules in the final products.
• The chemical mechanism can also be elucidated by examining
the kinetics and isotope effects under different pH conditions,
by altering the metal ions or other bound cofactors, by site-
directed mutagenesis of conserved amino acid residues, or by
studying the behaviour of the enzyme in the presence of
analogues of the substrate(s).
reaction, i.e., the sequence of chemical
steps that transform substrate into
product.
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