Tutorial #4: Enzyme Kinetic and Medicine This document was produced using the

LATEX typesetting language with the

Barry Linkletter Tufte-handout document class. Images
of proteins were created using UCSF
Chimera. Chemical diagrams were made
Many drugs act by inhibiting the action of enzymes. Sometimes this can be a
with ChemDoodle and further edited with
problem. Affinity Designer.

Enzymes and Drugs

Cytochrome P450 (cyp450) is a very important enzyme. It catalyzes the oxidation
of organic molecules by molecular oxygen. Oxygen will oxidize everything
on the surface of the Earth in about one million years, but this enzyme makes
that inevitable process much faster for specific molecules at specific chemical
sites within those molecules.
This enzyme is important is several biosynthetic pathways, but it is an
obsession of medicinal chemists due to its ability to oxidize many different
organic molecules. cyp450 is often one of the first steps in destroying drug
molecules in metabolism. The liver is famous for “detoxifying” the body
and one way it does this is through the activity of this enzyme. cyp450 is
responsible for the removal of drug activity in many cases – however, it may
also be responsible for the formation of toxic products that are harmful. Your
drug may be fine, but cyp450 may turn it into a poison. What works in the
test tube might be a disaster in the clinic.

Always be Prepared
To prepare for the quiz in this tutorial you should review the topic of enzyme
inhibition. And now is the time to know your amino acids.

Acetaminophen Toxicity
For example, the common drug acetaminophen, is mostly disposed of in our
bodies by conjugation with sugars in reactions catalyzed by several versions
of glucuronosyltransferase (EC 2.4.1.17).1 This makes the acetaminophen more 1
“The UDP-glucuronosyltransferases:
soluble in water and easily disposed of in urine. However, high concentrations Their role in drug metabolism and
detoxification”, A. Rowland, J.O. Miners,
of acetaminophen can be substrates for two versions of cyp450 – 2E1 and P.I. Mackenzie, J. Biochem. Cell. Biol.,
3A4. Both are fairly non-specific and act on small hydrophobic molecules. 2013, 45, 1121-32. https://doi.org/10.
1016/j.biocel.2013.02.019.
The site of oxidation is at the nitrogen. This leads to the production of
N-acetyl-p-benzoquinone imine (NAPQI), a toxic molecule.2 You can kill 2
“Pathways of acetaminophen metabolism
yourself with Tylenol, however you are more likely to just damage your liver at the therapeutic versus toxic doses”, L.L.
Mazaleuskaya, K. Sangkuhl, C.F. Thorn,
and suffer for the rest of your life. G.A. FitzGerald, R.B. Altman, T.E. Klein,
Today, we will consider an exercise calculating relative rates of these two Pharmacogenet. Genomics, 2015, 25, 416-
426. https://dx.doi.org/10.1097/FPC.
pathways at various concentrations of acetaminophen. 0000000000000150
 tutorial #4: enzyme kinetic and medicine 2

Figure 1: Some pathways for acetamin-

ophen in the liver. Glucuronidation via
Drug Interactions glucuronosyltransferase (EC 2.4.1.17) and
oxidation by molecular oxygen via cyp450
(EC 1.14.14.x). The relative rates of each
cyp450 is a major part of drug degradation in the liver. Figure 2 on the facing pathway will determine the harm to your
page shows several drugs that are known to be oxidized with the help of liver.
cyp450.3 There are many versions of cyp450 (more than 50 genes have been
identified) and it is often observed that one version has far more activity for a
particular drug than the others. Sometimes, another drug that is not oxidized 3
“Cytochrome P4502C9: an enzyme
as quickly by that version may, by chance, be an inhibitor of it. The presence of major importance in human drug
metabolism”, J.O. Miners, D.J. Birkett,
of the second drug slows removal of the first. Unless you are aware of this Br. J. Clin. Pharmacol., 1998, 45, 525-
interaction, you might be on your way to an overdose. 538. https://doi.org/10.1046/j.
1365-2125.1998.00721.x
Most inhibitors of cyp450 are competitive inhibitors. They occupy the
active site and block the substrate. A few are non-competitive inhibitors;
they bind elsewhere but reduce the reactivity of the catalytic enzyme-substrate
complex. Consider the crystal structure in figure 3. We see how a drug
has bound in the active site.4 This drug is actually inhibiting this version of 4
“Crystal Structure of CYP2C9*2 in
cyp450 by strongly binding to the active site and preventing oxygen from Complex with Losartan”, S.J. Parikh,
C.M. Evans, J.O. Obi, Q. Zhang,
associating with the iron of the heme. It won’t be oxidized, and it also K. Maekawa, K.C. Glass, M.B. Shah,
prevents any other organic molecules from entering (along with oxygen) to Molecular Pharmacology, 2020, 98, 529-539.
https://doi.org/10.1124/molpharm.
be hydroxylated. 120.000042
Today, we will explore an exercise in identifying competitive and non-
competitive inhibitors for cyp450.

Resources
Please share any useful resources that you find on the moodle forum.
tutorial #4: enzyme kinetic and medicine 3

Figure 2: Substrates for cytochrome P450.

The site of oxidation is indicated by the
arrows.3

Figure 3: Cytochrome P450 2C9 with

losartan.4 This is a mutant that binds
losartan in a way in which this enzyme
cannot perform the oxidation. It illustrates
how a drug might block cyp450.
 tutorial #4: enzyme kinetic and medicine 4

Tutorial #4 Plan

The Exercises
Today in tutorial we will enjoy a quiz that focuses on enzyme inhibition and
then explore enzyme kinetics as described above.

The Report
You are an employee of a major international pharma concern. Your job is to
evaluate a new drug for possible interactions with a common blood thinner.
If the concentrations of blood thinner increases unexpectedly the patient may
die of internal bleeding before they even know anything is wrong. We want
to know if our drug interferes with the action of cyp450 on warfarin.
For the report you will interpret the results of an enzyme experiment. Use
the data set that matches your student number. The data will include a series
of enzyme kinetics experiments with a drug being oxidized with catalysis by
cyp450. Then there will be a second set of data with the addition of a second
drug that acts as an inhibitor (we will assume that this second drug is oxidized
very slowly by ctyP450 and that process does not matter in the time frame
of or experiment).

• Present the structures of warfarin and your drug. Use your favourite
chemical drawing program. All drugs in the data sets are shown in figure 2
of this document.

• Using the conc. vs. time data from each experiment calculate the rate of
oxidation. Present a plot that shows all conc. vs. time plots in a single
graph.

• Plot rate vs. substrate concentration and report the KM and Vmax values
for warfarin with the version of cyp450 that you are investigating. Present
the Michaelis-Menten plot and any plots that you used to determine the
parameters.

• You will know the concentration of cyp450, so calculate and report kcat
and kcat / KM .

• Repeat the first two steps with the experimental data that includes the
presence of your new drug. Report the apparent KM and Vmax values for
warfarin in the presence of the new drug.

• Determine if the new drug is an inhibitor of the activity of cyp450 with

warfarin. If so, what kind of inhibitor – competitive or non-competitive?

• Calculate the Ki for the inhibitor. If the expected drug concentrations

in patients is well below the Ki , then the interaction may result in only a
small effect. We might still be able to sell it (with a long list of caveats and
lots of small print).
 tutorial #4: enzyme kinetic and medicine 5

The report should have a clear and well-organized appearance. All figures
should have legends (just look at the legends in this document for guidelines).
You might choose to present your final data in a table so that the executives
who make decisions don’t have to search through the report to quickly evaluate
app app
the conclusions. KM , Vmax , KM , Vmax , and Ki should be reported. Also
report the kcat and kcat /KM values for the uninhibited system.
The higher-ups at your behemoth corporation now have all the information
that they need to make a decision. Congratulations, you won’t be fired. . . at
least not today.

Python Challenges
We have had our first taste of using Python in the last tutorial. This time
we can type in all the data again and use the linear() function from the
previous tutorial to plot and curve fit each line. With the recorded slopes of
each run, you can calculate the rates (look at the units of the slopes – do you
need to change them?). Then you can type the numbers back in and use the
MMfit() function.
That seems tedious. Can we import the data from a text file? Can all this
be automated? Examine and steal the code provided in tutorial and change
it to work with your data set. Automate yourself out of your job and then
continue to be paid for it while you do other things. Computers are not
going to take your job – they are going to enable you to be paid for three
jobs at once. Stay on top of technology and you will stay on top of the heap.
 tutorial #4: enzyme kinetic and medicine 6

Data Tables
Below are the data tables. All data sets are available as text files in csv format.
Note: Look carefully to units. Mole, mmol, μmol, and nmol span orders
of magnitude that could describe the difference between you and the whole
solar system.

Data Set #0
Testing for the effect of amitriptyline on warfarin oxidation by cytochrome
P450. In all experiments the concentration of the enzyme was set to 1.0 nM

Table 1: Data Set #0: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.15 0.25 0.35 0.50 1.00 1.50 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 3.9 5.5 6.6 7.3 10.3 11.4
2.0 7.9 11.0 12.5 16.1 20.1 22.8
3.0 11.4 14.6 17.9 21.8 29.7 35.1
4.0 14.9 22.0 25.0 32.9 45.3 43.8
5.0 16.2 28.5 29.1 35.8 47.2 60.3
7.0 24.7 39.5 49.1 58.9 79.4 88.8
8.0 27.8 40.1 55.5 58.1 82.2 85.7

Table 2: Data Set #0: Amitriptyline

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.20 0.40 1.10 1.50 2.40 3.30 concentration of the potential inhibitor is
1.0 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 1.5 2.6 4.9 6.2 8.6 10.3
2.0 2.6 5.1 11.0 12.9 15.5 20.4
3.0 4.2 7.8 15.4 17.9 25.3 28.8
4.0 6.2 10.2 21.1 25.0 31.2 39.1
5.0 6.9 12.4 25.0 30.6 41.8 47.8
7.0 9.8 17.1 33.0 45.3 55.2 59.5
8.0 11.0 20.4 45.7 50.8 66.3 67.5
 tutorial #4: enzyme kinetic and medicine 7

Data Set #1
Testing for the effect of diclofenac on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 3: Data Set #1: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.12 0.20 0.28 0.40 0.80 1.20 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 4.0 6.7 6.9 10.0 12.3 13.7
2.0 8.3 12.7 16.7 19.0 24.2 29.6
3.0 12.9 18.8 21.3 27.4 39.2 38.3
4.0 17.1 24.1 28.3 35.4 44.9 53.9
5.0 20.4 32.5 34.5 45.8 66.1 71.2
7.0 29.4 46.3 51.2 64.0 82.0 90.2
8.0 33.9 54.0 64.4 76.1 97.2 115.9

Table 4: Data Set #1: Diclofenac

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.20 0.40 1.10 1.50 2.40 3.30 concentration of the potential inhibitor is
3.0 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 1.8 3.0 6.3 7.1 8.8 12.0
2.0 3.6 6.0 13.5 15.5 19.9 23.6
3.0 4.7 9.4 19.6 22.7 27.7 33.3
4.0 6.8 13.5 24.7 28.1 35.3 40.6
5.0 8.7 15.2 33.5 41.6 49.9 56.7
7.0 11.7 22.3 44.2 48.5 69.7 75.3
8.0 13.5 22.4 53.3 61.4 74.5 86.6
 tutorial #4: enzyme kinetic and medicine 8

Data Set #2
Testing for the effect of fluoxetine on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 5: Data Set #2: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.27 0.45 0.63 0.9 1.8 2.7 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 5.6 7.5 10.2 11.1 15.7 16.6
2.0 11.8 17.2 19.5 26.0 32.2 37.2
3.0 16.4 22.7 32.1 38.0 48.7 49.9
4.0 23.2 29.5 43.3 52.6 64.8 71.1
5.0 29.6 39.7 45.1 65.2 81.3 93.5
7.0 37.5 51.0 69.3 84.8 117.2 119.4
8.0 44.4 61.5 83.0 87.0 129.8 134.7

Table 6: Data Set #2: Fluoxetine

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.09 0.18 0.45 0.63 0.99 1.35 concentration of the potential inhibitor is
1.0 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 0.76 1.4 2.7 3.3 4.4 4.6
2.0 1.3 2.9 4.8 6.8 8.3 9.6
3.0 2.0 4.0 7.3 10.4 13.5 15.3
4.0 2.7 5.8 11.4 12.0 17.7 19.6
5.0 3.8 7.2 13.1 16.8 20.7 24.5
7.0 5.0 9.4 17.3 25.0 29.9 33.5
8.0 5.8 10.1 19.7 26.3 30.2 40.4
 tutorial #4: enzyme kinetic and medicine 9

Data Set #3
Testing for the effect of mefenamic acid on warfarin oxidation by cytochrome
P450. In all experiments the concentration of the enzyme was set to 1.0 nM

Table 7: Data Set #3: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.24 0.40 0.56 0.80 1.60 2.40 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 3.5 5.0 6.6 7.3 11.0 10.5
2.0 6.8 10.7 11.7 15.6 20.3 22.7
3.0 11.0 15.1 19.7 22.9 29.5 34.3
4.0 12.6 18.0 26.8 30.6 39.6 49.1
5.0 18.7 25.8 31.7 39.4 53.3 53.3
7.0 25.5 37.4 40.1 50.2 73.5 73.1
8.0 25.3 40.4 44.8 59.5 80.1 83.1

Table 8: Data Set #3: Mefenamic acid

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.10 0.20 0.40 0.60 0.90 1.20 concentration of the potential inhibitor is
1.2 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 0.5 0.8 1.7 2.1 2.8 2.9
2.0 0.8 1.7 3.4 3.8 5.4 5.6
3.0 1.3 2.6 4.7 6.7 8.4 9.8
4.0 1.6 3.1 7.2 8.0 9.9 12.9
5.0 2.3 4.1 8.8 9.4 14.2 13.9
7.0 3.1 6.4 12.0 15.7 16.5 22.6
8.0 3.9 7.0 12.4 17.9 22.6 22.7
 tutorial #4: enzyme kinetic and medicine 10

Data Set #4
Testing for the effect of piroxicam on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 9: Data Set #4: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.60 1.00 1.40 2.00 4.00 6.00 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 5.8 8.6 11.0 12.6 17.1 19.4
2.0 10.3 17.2 20.4 23.6 36.0 34.2
3.0 15.6 24.1 31.0 39.7 45.5 54.1
4.0 23.4 32.2 39.8 48.6 65.9 76.3
5.0 28.2 40.4 48.8 60.0 88.1 92.9
7.0 38.1 62.1 64.4 90.8 107.2 137.1
8.0 48.1 59.5 86.2 106.1 139.2 143.1

Table 10: Data Set #4: Piroxicam

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.10 0.20 0.40 0.60 0.90 1.20 concentration of the potential inhibitor is
1.2 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 2.2 4.2 8.8 9.5 13.4 13.3
2.0 4.1 8.0 15.5 18.8 25.2 29.4
3.0 7.1 11.1 22.5 33.0 37.9 44.3
4.0 8.7 16.6 31.9 41.8 52.4 64.8
5.0 11.1 22.5 42.1 52.1 59.7 72.4
7.0 15.9 28.5 62.3 64.4 91.6 111.4
8.0 17.9 33.6 71.7 84.0 108.9 115.7
 tutorial #4: enzyme kinetic and medicine 11

Data Set #5
Testing for the effect of phenytoin on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 11: Data Set #5: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.63 1.05 1.47 2.1 4.2 6.3 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 2.7 3.7 4.5 5.8 7.4 8.5
2.0 5.2 6.6 8.2 11.1 15.3 15.5
3.0 7.1 11.7 12.9 15.1 21.1 25.5
4.0 10.6 14.6 16.9 23.3 29.2 29.9
5.0 11.4 16.5 23.6 29.4 35.9 39.5
7.0 18.1 25.6 28.1 37.3 49.5 57.5
8.0 20.9 29.9 38.5 43.7 57.1 61.5

Table 12: Data Set #5: Phenytoin

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.21 0.42 1.05 1.47 2.31 3.15 concentration of the potential inhibitor is
1.1 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 0.35 0.77 1.4 1.7 2.0 2.3
2.0 0.80 1.5 2.8 3.7 4.1 4.8
3.0 1.2 2.2 3.9 5.0 6.5 7.2
4.0 1.4 2.8 5.2 7.6 9.5 9.9
5.0 2.0 3.4 6.5 9.4 10.3 11.6
7.0 2.9 4.5 10.1 11.4 16.8 18.3
8.0 2.8 5.8 11.7 14.1 17.4 19.6
 tutorial #4: enzyme kinetic and medicine 12

Data Set #6
Testing for the effect of seratrodast on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 13: Data Set #6: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.93 1.55 2.17 3.10 6.20 9.30 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 3.6 4.6 6.4 7.2 9.3 11.5
2.0 6.4 9.2 11.0 15.5 18.4 21.8
3.0 9.4 14.8 17.2 22.5 30.9 29.2
4.0 14.3 18.4 22.2 30.6 38.3 42.1
5.0 18.0 23.7 32.1 38.5 50.8 52.4
7.0 24.0 30.7 44.5 48.5 65.4 82.5
8.0 27.2 38.6 43.3 57.5 75.3 81.8

Table 14: Data Set #6: Seratrodast

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.31 0.62 1.55 2.17 3.41 4.65 concentration of the potential inhibitor is
1.1 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 0.65 1.1 2.1 2.7 3.6 4.2
2.0 1.2 2.4 4.6 6.0 7.8 8.0
3.0 1.8 3.5 6.3 8.4 10.6 12.2
4.0 2.4 5.0 8.9 12.0 15.7 15.6
5.0 3.3 5.9 12.2 13.6 17.7 19.6
7.0 4.4 7.4 16.5 19.3 24.5 26.8
8.0 5.2 10.0 19.0 22.0 26.4 34.0
 tutorial #4: enzyme kinetic and medicine 13

Data Set #7
Testing for the effect of tenoxicam on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 15: Data Set #7: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
2.70 4.50 6.30 9.00 18.0 27.0 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 5.1 8.8 11.0 13.1 15.8 20.3
2.0 10.7 17.8 21.0 23.8 32.3 40.2
3.0 15.9 24.8 31.6 38.7 53.2 55.3
4.0 24.2 35.4 39.0 48.1 61.0 70.7
5.0 26.1 40.3 46.5 58.0 76.6 88.7
7.0 41.5 58.6 67.9 86.3 107.4 133.2
8.0 42.4 66.5 78.7 102.2 125.7 149.2

Table 16: Data Set #7: Tenoxicam

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.90 1.80 4.50 6.30 9.90 13.5 concentration of the potential inhibitor is
2.0 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 1.0 2.1 4.3 5.0 6.0 7.1
2.0 2.3 4.0 7.6 9.6 13.8 15.5
3.0 3.3 6.1 11.6 15.7 20.7 22.3
4.0 4.2 8.5 16.5 20.6 23.4 27.5
5.0 5.6 10.5 22.5 26.8 32.7 37.3
7.0 7.4 13.6 26.1 37.5 49.5 47.4
8.0 9.5 16.4 31.4 41.8 54.9 54.4
 tutorial #4: enzyme kinetic and medicine 14

Data Set #8
Testing for the effect of tolbutamide on warfarin oxidation by cytochrome
P450. In all experiments the concentration of the enzyme was set to 1.0 nM

Table 17: Data Set #8: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
2.25 3.75 5.25 7.50 15.0 22.5 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 4.9 7.2 10.3 12.1 13.7 18.2
2.0 10.9 13.7 20.3 21.0 32.1 31.1
3.0 15.2 24.4 26.3 31.5 45.4 53.4
4.0 19.6 31.9 37.4 49.8 62.1 71.8
5.0 24.3 41.7 44.8 58.3 73.3 82.8
7.0 38.3 58.4 68.7 76.4 104.4 122.2
8.0 45.0 60.7 67.9 84.4 131.1 143.8

Table 18: Data Set #8: Tolbutamide

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.75 1.5 3.75 5.25 8.25 11.25 concentration of the potential inhibitor is
2.2 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 0.95 1.7 3.5 3.9 5.2 5.6
2.0 1.9 3.2 6.5 8.8 11.7 12.1
3.0 2.7 4.9 10.6 11.6 17.7 18.9
4.0 3.8 7.2 12.9 15.7 19.7 23.6
5.0 4.9 8.2 18.7 22.8 26.4 28.8
7.0 7.0 11.6 26.3 29.0 39.6 44.3
8.0 6.8 14.6 27.6 34.1 43.1 53.0
 tutorial #4: enzyme kinetic and medicine 15

Data Set #9
Testing for the effect of torsemide on warfarin oxidation by cytochrome P450.
In all experiments the concentration of the enzyme was set to 1.0 nM

Table 19: Data Set #9: Oxidation of

Conc. of warfarin /mmol L−1 warfarin with cyp450. No potential
0.90 1.50 2.10 3.00 6.00 9.00 inhibitor is present.

time (min) Conc. of product /μmol L−1

1.0 4.0 5.3 6.6 8.3 11.9 13.0
2.0 7.3 12.4 13.6 18.0 21.3 26.6
3.0 13.1 17.0 23.1 25.4 38.2 38.6
4.0 16.2 23.2 26.6 33.6 47.9 52.3
5.0 20.6 26.6 35.6 43.3 53.1 67.0
7.0 26.6 38.1 45.4 57.4 78.7 97.3
8.0 33.2 50.3 57.9 76.2 98.9 100.6

Table 20: Data Set #9: Torsemide

Conc. of warfarin /mmol L−1 interaction with warfarin metabolism. The
0.55 1.10 2.75 3.85 6.05 8.25 concentration of the potential inhibitor is
1.5 mmol L −1
time (min) Conc. of product /μmol L−1
1.0 1.7 3.2 6.1 6.9 10.0 11.5
2.0 3.2 5.5 11.8 13.9 16.6 22.8
3.0 5.1 8.7 18.4 19.5 30.0 31.9
4.0 6.9 10.7 21.6 26.8 38.9 38.1
5.0 8.5 15.0 29.3 32.7 42.1 52.8
7.0 10.5 21.0 40.0 50.9 69.7 67.7
8.0 12.7 24.5 47.0 56.4 77.3 83.6