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Lab Report Bio462 - Exp 3

This document summarizes experiments 3 and 4 from the Biochemistry course BIO462. In experiment 3, students determined the optimum parameters for enzyme activity by investigating the effect of enzyme concentration, substrate concentration, and pH on the rate of reaction. The results showed inconsistencies compared to expected trends. In experiment 4, students determined the activity of the enzyme acetylcholinesterase and generated a standard curve for bovine serum albumin (BSA) concentration, which also showed errors. References are provided for expected results on the effect of concentration on enzyme activity.

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126 views

Lab Report Bio462 - Exp 3

This document summarizes experiments 3 and 4 from the Biochemistry course BIO462. In experiment 3, students determined the optimum parameters for enzyme activity by investigating the effect of enzyme concentration, substrate concentration, and pH on the rate of reaction. The results showed inconsistencies compared to expected trends. In experiment 4, students determined the activity of the enzyme acetylcholinesterase and generated a standard curve for bovine serum albumin (BSA) concentration, which also showed errors. References are provided for expected results on the effect of concentration on enzyme activity.

Uploaded by

Ms. Nisa
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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BIO462 - BIOCHEMISTRY

Experiment 3 and 4 (Enzymology)


Date of Experiment: 20/11/2023 (Monday)
Class : AS2571A
Lecturer: Prof. Dr. Farida Zuraina Mohd Yusof

Group Member:
NO NAME STUDENT ID

1 NURFARISYA ATIRAH BINTI AHMAD FAUZI 2023492164


2 NUR IMANINA JEMILA BINTI MOHD HISHAM 2023225404
3 WAN NURUL NAJWA BINTI WAN KHAIRUZZAMAN 2023600974
4 ATIKAH SYAZANA BINTI ABD HALIM 2023686716
5 SITI AISHAH BINTI AZLEE SHAH 2023213918
6 NURAIN NISA BINTI MOHD SATAR 2023660438
RESULTS :

Experiment 3 : Enzymology (Determination of Optimum Parameter)

A. Calibration and setting for spectrophotometer

Reading 1: 0.367
Reading 2: 0.311
Reading 3: 0.301

Show that the spectrophotometer is calibrated because each three readings is


approximately.

B. The effect of enzyme concentration on the rate of reaction

Cuvette OD OD OD Mean +- S.D


(replicate 1) (replicate 2) (replicate 3)

1 0.470 0.437 0.430 0.441

2 0.395 0.195 0.331 0.307

3 0.260 0.106 0.215 0.174

4 0.140 0.097 0.206 0.148

C. The effect of substrate concentration on the rate of reaction

Cuvette OD OD OD Mean +- S.D


(replicate 1) (replicate 2) (replicate 3)

1 0.100 0.105 0.100 0.102

2 0.110 0.112 0.113 0.112

3 0.170 0.173 0.170 0.171

4 0.250 0.248 0.254 0.251

5 0.394 0.394 0.369 0.386

6 0.594 0.604 0.600 0.599

7 0.835 0.830 0.837 0.834


D. The effect of pH on enzyme activity

Cuvette OD OD OD Mean +- S.D


(replicate 1) (replicate 2) (replicate 3)

1 0.134 0.178 0.185 0.17

2 0.157 0.165 0.171 0.16

3 0.134 0.156 0.158 0.15

4 0.328 0.421 0.426 0.41

5 0.220 0.279 0.339 0.28

6 0.298 0.265 0.252 0.27

Graph I :

Graph 1.0 : Obtained results


Graph 1.1 : Expected results
(Libretexts, 2021)

The obtained result between enzyme concentration (X-axis) and rate of reaction (Y-axis)
sloped down horizontally differ from expected results. This may happened due to
interference during performing experiments. As enzyme concentration increase in the
solution, the rate of reaction are supposed to be increasing then flattened out as the
substrate has finished to be react.

Graph II :
Graph III :
Experiment 4 : Enzymology (Determination of Enzyme’s activity)

A. Absorbance of BSA

Concentration OD OD OD Mean (± S.D)


of BSA (replicate 1) (replicate 2) (replicate 3)

0.2 mg/ml 0.002 0.091 0.020 0.038

0.4 mg/ml 0.027 0.006 0.120 0.051

0.6 mg/ml 0.015 0.017 0.003 0.012

0.8 mg/ml 0.025 0.037 0.024 0.029

1.0 mg/ml 0.047 0.050 0.039 0.045

B. Enzyme activity

OD OD OD Mean (± S.D)
(replicate 1) (replicate 2) (replicate 3)

Acetylcholinesterase 0.120 0.125 0.121 0.122


The expected result of standard curve of BSA should be increasing but from the experiment
showed error in the standard curve.Inaccurate readings of the absorbance might be caused by
the spectrophotometer's instrumentation being inaccurate or being insufficiently sensitive to
detect absorbance accurately.

REFERENCES :

1. Libretexts. (2021, January 15). 19.5: Effect of Concentration on Enzyme Activity.

Chemistry LibreTexts.

https://chem.libretexts.org/Courses/Saint_Francis_University/Chem_114%3A_Human_C

hemistry_II_(Muino)/19%3A_Enzymes_and_Vitamins/19.05%3A_Effect_of_Concentrat

ion_on_Enzyme_Activity

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