Experiment No.

3
Principles of Enzyme Catalysis
June 28, 2012
Biochemistry
Abstract
Cells undergo many complicated and time consuming biological processes; however, to
significantly reduce the time consumed for such reactions enzyme catalysis is used.
Rate of enzyme catalyzed reactions vary significantly with temperature, time, pH, and
the concentrations of substrate and enzymes. Knowing the relationship of these would
be of great help in the use of enzymes in the clinical setting for diagnosis, therapy or
even for industrial pusposes.
The experiment aims to determine the correlation of time with the rate of reaction
Using a spectrophotometer (Ultrospec 1100 pro UV/Vis) set at 500 nm, the absorbance
was read from the longest catalyzed reaction and the shortest catalyzed reaction every
half minute of interval.
Background, objectives, methods, results and conclusion (word count)
Results (2nd page)

Assa
y

Assay

Volume, mL
Diluted
Con

Rxn

Abs500n

Soluti

Phospha

Soluti

Soluti

on
2.0

te Buffer
0.2

on
--

on
--

--

0.0

2.0

--

0.2

--

0.663

0.5

2.0

--

--

0.2

0.536

Tube

[peroxid
e],
Molarity
-0.00139
2
0.00112

1.0

2.0

--

--

0.2

0.472

1.5

2.0

--

--

0.2

0.412

2.0

2.0

--

--

0.2

0.355

2.5

2.0

--

--

0.2

0.304

3.0

2.0

--

--

0.2

0.246

5
0.00099
1
0.00086
5
0.00074
5
0.00063
8
0.00051
6

Enzyme Catalysis
0.001600
0.001400
0.001200
0.001000
Peroxide [M]

f(x) = - 0x + 0
R = 0.98

0.000800

Linear ()

0.000600
0.000400
0.000200
0.000000
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5
Time [Minutes]

Interpretation and Discussion of Results

By determining the correlation of the trendline with the actual values obtained in the
experiment
Conclusion

References
Murray, Robert K., D. K. Granner and V. W. Rodwell. Harpers Illustrated
Biochemistry, 27th Ed. McGraw-Hill Companies, Inc., Singapore. 2006.
EDVOTEK, The Biotechnology Education Company Manual. Advance Placement
Biology Lab 2Principles of Enzyme Catalysis. 2008.
Appendix
Appendix A
Determination of the peroxide concentration in the reaction tubes
[Peroxide]rxn tube= ( Absorbance / ) * ( 2.2 mL / 0.2 mL )
[Peroxide]rxn tube= ( Absorbance / ) * 11
where,
= molar extinction coefficient for the assay system
In this experiment,
= 5.24x103 cm-1 M-1
[Peroxide]rxn tube= ( Absorbance / 5.24x103 cm-1 M-1 ) * 11
Appendix B
Determination of the rate of enzymatic reaction from the slope of the plot obtained
m = [peroxide] / minute
correlating the values via linear regression of [peroxide] vs time (min),

= -0.9876

= 0.00131

= -2.75x10-4

the equation for the best fit line,

y = - 0.000275x + 0.00131
rounding off,
y = - 0.0003x + 0.0013
R2=0.9753
Rate = 2.75x10-4 M/min