... for example, the reaction A → B →
C. Usually, equations in differential or algebraic form are fitted to indi-
vidual data sets, A, B, and C and a set of parameter estimates obtained.
26 TOOLS AND ... constant, and
only k
2
and k
3
floated. After preliminary parameter estimates are obtained
in this fashion, these parameters should be fixed as constants and the
remaining parameters estimat...
... is appropriate. When more
than two values are compared, a one-way analysis of variance (ANOVA),
TABLE 4 .2 Rate of Hydration of Fumarate to Malate by Fumarase at various
Substrate Concentrations
a
Velocity ... phenomenological irreversible inhibition mechanisms are
discussed in turn.
APPLICATIONS 67
TABLE 4.3 Estimates of the Catalytic Parameters for the Fumarase-Catalyzed
hydration of F...
... enzyme substrate complex EAB (K
AB
), and
the enzyme mass balance are, respectively,
v = k
cat
[EAB] (7 .21 )
K
A
s
=
[E] [A]
[EA]
K
AB
=
[EA][B]
[EAB]
(7 .22 )
[E
T
] = [E] + [EA] + [EAB] (7 .23 )
Normalization ... as
v
V
max
=
[B]
K
+ [B]
(7. 12)
where
V
max
=
V
max
[A]
K
BA
+ [A]
(7.13)
and
K
=
K
A
s
K
AB
+ K
AB
[A]
K
BA
+ [A]
=
K
AB
(K
A
s
+ [A] )
K
BA
+ [A]
(7.1...
... exponential term, and the shape of the
curve approaches that of a straight line. Valuable information can be
gained from analysis of the early and late stages of this reaction.
11 .2. 1 Early Stages ... plot for an interfa-
cial enzyme.
various parameters in Eq. (10.9) on the velocity of a reaction catalyzed
by an interfacial enzyme. As the enzyme interface dissociation constant
incre...
... derivation
of an enzyme catalysis model. A steady-state approximation can also be
used to obtain the rate equation for an enzyme- catalyzed reaction.
3 .2. 2 Steady-State Model
The main assumption made in ... scattered randomly about zero. A large positive value of the
SCC is indicative of a systematic deviation of the model from the data.
PRACTICAL EXAMPLE 55
TABLE 3 .2 Average and...
... major subclasses of mechanism-based inhibition.
Alternate substrates are processed by an enzyme s normal catalytic
pathway to form a stable covalent enzyme inhibitor intermediate, such
as an acyl -enzyme ... enzyme A.
12. 3 .2 Kinetic Characterization of Stability
Decreases in the activity of an enzyme as a function of time, at dif-
ferent temperatures, are shown in Table 12. 2...
... Groutas
et al., 1998; Macchia et al., 20 00; Clemente et al., 20 01). Amino acid
analysis of both native and inactivated enzyme can identify which amino
acid is modified (for examples see Pochet et al., ... k
on
and K
i
of an alter-
nate substrate. Addition of an alternate substrate inhibitor to an enzyme
assay results in an exponential decrease in rate to some final steady-
state turnove...
... lysine–proline–alanine–glutamic acid–phenylalanine–
phenylalanine (NO
2
)–alanine–leucine.
c
ss2, substrate consisting of the peptide leucine–serine–phenylalanine (NO
2
)–norleucine–leucine–
methyl ... mean of a minimum of two deter-
minations ± standard deviation.
b
ss1, substrate consisting of the peptide lysine–proline–alanine–glutamic acid–phenylalanine–
phenylalanine (NO
2
)–alan...