... Groutas
et al., 19 98; Macchia et al., 2000; Clemente et al., 2001). Amino acid
analysis of both native and inactivated enzyme can identify which amino
acid is modified (for examples see Pochet et al., ... 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 turnover of substra...
... TOOLS AND TECHNIQUES OF KINETIC ANALYSIS
1.4 ACID–BASE CHEMICAL CATALYSIS
Many homogeneous reactions in solution are catalyzed by acids and bases.
ABr
¨
onsted acid is a proton donor,
HA + H
2
O ... 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 TOOL...
... 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 ... K
∗
s
correspond, respectively, to apparent enzyme maxi-
mum velocity and apparent enzyme substrate dissociation constant at a
particular pH. For the model above, V
∗
max
= V
ma...
... cooperativity, which can have noninteger values.
Estimates of the parameters k
and n are obtained using standard non-
linear regression procedures available in most modern graphical software
packages. ... constant,
Eq. (7.11) can be expressed 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
+...
... 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
increas...
... expressed as
d [A]
dt
=−k
r
[A] (1 .8)
Integration of Eq. (1 .8) for the boundary conditions A = A
0
at t = 0and
A = A
t
at time t,
A
t
A
0
d [A]
[A]
=−k
r
t
0
dt(1.9)
yields the integrated rate equation ... specifically disclaim any implied
warranties of merchantability or fitness for a particular purpose. No warranty may be created or
extended by sales representatives...
... statistic:
χ
2
=
n
i=1
(y
i
−ˆy
i
)
2
s
2
i
=
n
i=1
w
i
(y
i
−ˆy
i
)
2
(1.125)
Consider a typical experiment where the value of a dependent variable is
measured several times at a particular value of the independent variable.
From these repeated determinations, a mean and variance ... Exact Analytical Solution (Non-Steady-State
Approximation)
Exact analytical solutions for the rea...
... treatment above assumes that there are no differences in heat capac-
ity between native and denatured states of an enzyme and that the heat
capacity remains constant throughout the temperature ... stability, enzyme solutions are
incubated at a particular temperature and aliquots removed at the appro-
priate times. Enzyme activity in these samples is then measured at the
enzyme s temp...
... less than either valine
and serine mutants, which had comparable catalytic constants. Compared
to valine and serine, alanine has the smallest van der Waals volume and
accessible surface area. Therefore, ... mean of a minimum of two determi-
nations ± standard deviation.
b
ss1, substrate consisting of the peptide lysine–proline–alanine–glutamic acid–phenylalanine–
phenylalanine (NO
2
)–alani...