3. Pyrozole has been proposed as a possible nontoxic inhibitor of LADH-catalyzed ethanol oxidation. Its kinetics have been studied and, in 2 separate experiments, the velocity of the reaction was measured as a function of [C₂H5OH] and of [NAD+]. The data is given in the tables below. [C₂H5OH] varies, [LADH]= 4µg/ml, [NAD+] = 350 µM, pH=7.4, 23.5C [C₂H5OH] Vo (relative units) Vo (relative units) 1x10-5 M pyrazole M No pyrazole 2.1 1.8 1.5 1.35 0.008 0.002 0.001 0.00067 1.7 1.0 Vo (relative units) No pyrazole 0.00011 1.7 0.000056 1.4 0.000033 1.3 0..000017 0.95 0.69 0.52 [NAD+] varies, [LADH]= 4µg/ml, [C₂H5OH]= 5 mM, pH=7.4, 23.5C [NAD+] Vo (relative units) 4x10-5 M pyrazole 0.85 0.71 0.63 0.47 a. Plot the data as Lineweaver-Burke plots on two separate graphs. b. Determine the Km values for [C₂H5OH] and of [NAD+]. (You can't find Vmax because the exact units of Vo are not provided) c. What type of inhibition is exhibited by pyrazole against C₂H5OH is a substrate? Describe what is going on in this type of inhibition.
3. Pyrozole has been proposed as a possible nontoxic inhibitor of LADH-catalyzed ethanol oxidation. Its kinetics have been studied and, in 2 separate experiments, the velocity of the reaction was measured as a function of [C₂H5OH] and of [NAD+]. The data is given in the tables below. [C₂H5OH] varies, [LADH]= 4µg/ml, [NAD+] = 350 µM, pH=7.4, 23.5C [C₂H5OH] Vo (relative units) Vo (relative units) 1x10-5 M pyrazole M No pyrazole 2.1 1.8 1.5 1.35 0.008 0.002 0.001 0.00067 1.7 1.0 Vo (relative units) No pyrazole 0.00011 1.7 0.000056 1.4 0.000033 1.3 0..000017 0.95 0.69 0.52 [NAD+] varies, [LADH]= 4µg/ml, [C₂H5OH]= 5 mM, pH=7.4, 23.5C [NAD+] Vo (relative units) 4x10-5 M pyrazole 0.85 0.71 0.63 0.47 a. Plot the data as Lineweaver-Burke plots on two separate graphs. b. Determine the Km values for [C₂H5OH] and of [NAD+]. (You can't find Vmax because the exact units of Vo are not provided) c. What type of inhibition is exhibited by pyrazole against C₂H5OH is a substrate? Describe what is going on in this type of inhibition.
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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![**Transcription for Educational Website**
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### Pyrazole as a Nontoxic Inhibitor of LADH-Catalyzed Ethanol Oxidation
Pyrazole has been studied as a potential nontoxic inhibitor for LADH-catalyzed ethanol oxidation. In two separate experiments, the reaction velocity was measured as a function of ethanol concentration \([C_2H_5OH]\) and NAD\(^+\). The data is presented below.
#### Experiment 1: [C₂H₅OH] Variation
- **Conditions**: \([LADH] = 4 \, \mu g/ml, [NAD^+] = 350 \, \mu M, \, \text{pH}=7.4, \, 23.5°C\)
| \([C_2H_5OH]\) (M) | \(V_o\) (relative units) No pyrazole | \(V_o\) (relative units) \(1 \times 10^{-5} \, M \) pyrazole |
|----------------|-----------------------------------|--------------------------------------|
| 0.008 | 2.1 | 1.7 |
| 0.002 | 1.8 | 1.0 |
| 0.001 | 1.5 | 0.69 |
| 0.000067 | 1.35 | 0.52 |
#### Experiment 2: [NAD⁺] Variation
- **Conditions**: \([LADH] = 4 \, \mu g/ml, [C_2H_5OH] = 5 \, mM, \, \text{pH}= 7.4, \, 23.5°C\)
| \([NAD^+]\) | \(V_o\) (relative units) No pyrazole | \(V_o\) (relative units) \(4 \times 10^{-5} \, M \) pyrazole |
|----------------|-----------------------------------|--------------------------------------|
| 0.00011 | 1.7 | 0.85 |
| 0.000056 | 1.4 | 0.71 |
| 0.000033 | 1.3 | 0.](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Ff123c09d-ccbd-492d-9b0f-c8892232526f%2Ffe10f1d3-3663-4f12-baf8-724d7e358d1d%2Fbaqkq9c_processed.png&w=3840&q=75)
Transcribed Image Text:**Transcription for Educational Website**
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### Pyrazole as a Nontoxic Inhibitor of LADH-Catalyzed Ethanol Oxidation
Pyrazole has been studied as a potential nontoxic inhibitor for LADH-catalyzed ethanol oxidation. In two separate experiments, the reaction velocity was measured as a function of ethanol concentration \([C_2H_5OH]\) and NAD\(^+\). The data is presented below.
#### Experiment 1: [C₂H₅OH] Variation
- **Conditions**: \([LADH] = 4 \, \mu g/ml, [NAD^+] = 350 \, \mu M, \, \text{pH}=7.4, \, 23.5°C\)
| \([C_2H_5OH]\) (M) | \(V_o\) (relative units) No pyrazole | \(V_o\) (relative units) \(1 \times 10^{-5} \, M \) pyrazole |
|----------------|-----------------------------------|--------------------------------------|
| 0.008 | 2.1 | 1.7 |
| 0.002 | 1.8 | 1.0 |
| 0.001 | 1.5 | 0.69 |
| 0.000067 | 1.35 | 0.52 |
#### Experiment 2: [NAD⁺] Variation
- **Conditions**: \([LADH] = 4 \, \mu g/ml, [C_2H_5OH] = 5 \, mM, \, \text{pH}= 7.4, \, 23.5°C\)
| \([NAD^+]\) | \(V_o\) (relative units) No pyrazole | \(V_o\) (relative units) \(4 \times 10^{-5} \, M \) pyrazole |
|----------------|-----------------------------------|--------------------------------------|
| 0.00011 | 1.7 | 0.85 |
| 0.000056 | 1.4 | 0.71 |
| 0.000033 | 1.3 | 0.
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