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Enzyme Lab: Virtual Enzyme Kinetics

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Enzyme Lab: Virtual Enzyme Kinetics

Khoa Nguyen        

40078865

Summary (conclusions):

An enzyme (ref. PR/05-210-101) has been assigned.

The optimum pH for the enzyme to function is 7

[pic 1]

The azide inhibits the catalytic process of the enzyme because it acts as a noncompetitive inhibition. To explain,  azide binds to an enzyme somewhere other than the active site. Then, enzyme's three-dimensional structure is changed; its active site can still bind substrate with the usual affinity, but is no longer in the optimal arrangement to stabilize the transition state and catalyze the reation. The dissociation constant for an enzyme -inhibitor complex:

[pic 2]

From: Isbu.ac.uk, Enzyme Inhibition, 2014

The precision of the measurements for the rates was seen by a standard deviation of 0.082 for the enzyme without the inhibitor. For the measurements taken for the enzyme and inhibitor the precision of the tests were seen by a 0.032 standard deviation. The precision did vary with the different rates seen because of the presence or lack of presence of the inhibitor.

Results

Without Inhibitors

Solute Effect

pH Effects

Precision

[S] (nM)

Vo (umol/min)

pH

Vo (umol/min)

Trail

Vo (umol/min)

0.3

0.3863

4

Too slow

1

3.34

50

4.5464

5

0.4248

2

3.30

0.43

0.7837

6

2.425

3

3.16

1.28

1.6646

7

4.2934

4

3.24

4.27

3.1459

8

4.266

5

3.16

12.8

4.234

9

2.1431

Variable

Constant

Variable

Constant

Variable

Constant

[S]

pH, [I], Volume

pH

[S], [I], Volume

[S], [I], pH, Volume

Estimated Km: 2.13 🡪 Sample calculation: S ((V50/V0.1) – 1) = 0.2((4.55/0.39) -1) = 2.13

Actual Km: 2.26 +/- 0.178

Vmax: 4.77 +/- 0.194

[pic 3]

Precision: Mean of 3.240 μmol/min with a Standard Error of 0.082 μmol/min

[pic 4]

With Inhibitors

Solute Effect

pH Effects

[S] (nM)

Vo (umol/min)

pH

Vo (umol/min)

0.43

Too slow

6

0.722

0.75

0.2723

6

0.7244

1.28

0.3852

6

0.6673

4.27

0.7214

6

0.7173

12.8

0.9428

7

1.2274

6.4

0.7617

7

1.2167

10

0.882

7

1.2021

7

1.2209

7

Too slow

7

1.2144

7

1.1416

Variable

Constant

Variable

Constant

[S]

pH, [I], Volume

pH

[S], [I], Volume

...

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