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Lab 8 - Result and Calculation
Lab 8 - Result and Calculation
0 RESULTS
Table 7.1: The values of absorbance Optical Density at different glucose concentration.
Glucose concentration (g/L) Absorbance Optical Density (OD) (nm)
0 0
600 3.57
700 2.98
900 4.87
6
Absorbance Optical Density (OD)(nm)
y = 0.0052x + 0.0072
5 R² = 0.9471
0
0 100 200 300 400 500 600 700 800 900 1000
Glucose concentration (g/L)
Figure 7.1: The standard curve of absorbance Optical Density (OD) (nm) against glucose
concentration (g/L)
EFFECT OF TEMPERATURE:
Table 7.2: The values of Absorbance Optical Density (OD) (nm) at different temperature.
Temperature (oC) Absorbance Optical Density (OD)
30 8.72
40 7.46
50 9.05
60 8.65
70 9.78
12
Absorbance Optical Density (OD) (nm)
10
0
0 10 20 30 40 50 60 70 80
Temperature (oC)
Figure 7.2: The values of Absorbance Optical Density (OD) (nm) against temperature (oC)
Table 7.3: The values of enzyme activity at different temperatures.
Temperature Absorbance Optical Glucose concentration, Glucose Enzyme activity,
(oC) Density (OD) X (g/mL) released (mol) V (mol/min)
0.0000012
0.000001
Enzyme activity, V (mol/min)
0.0000008
0.0000006
0.0000004
0.0000002
0
0 10 20 30 40 50 60 70 80
Temperature (oC)
Table 7.4: The values of Absorbance Optical Density (OD) (nm) at different pH values.
pH value Absorbance Optical Density (OD)
5 10.58
6 8.13
7 9.47
8 9.90
12
10
Absorbance Optical Density (OD)(nm)
0
0 1 2 3 4 5 6 7 8 9
pH Values
Figure 7.4: The values of Absorbance Optical Density (OD) (nm) against pH values
Table 7.5: The values of enzyme activity at different pH values.
pH Absorbance Optical Glucose concentration, Glucose Enzyme activity,
Value Density (OD) X (g/mL) released (mol) V (mol/min)
0.0000012
0.000001
Enzyme activity, V (mol/min)
0.0000008
0.0000006
0.0000004
0.0000002
0
0 1 2 3 4 5 6 7 8 9
pH Values
Table 7.6: The values of Absorbance Optical Density (OD) (nm) at different substrate
concentration.
Substrate Concentration (%) Absorbance Optical Density (OD)
0.5 10.57
1.5 7.80
2.0 9.32
2.5 8.88
3.0 7.92
12
10
Absorbance Optical Density (OD) (nm)
0
0 0.5 1 1.5 2 2.5 3 3.5
Substrate Concentration (%)
Figure 7.6: The values of Absorbance Optical Density (OD) (nm) against substrate concentration
Table 7.7: The values of enzyme activity at different substrate concentration and the data for
Michaelis – Menton
1.5 7.80 0.00150 8.33 × 10-6 8.33 × 10-7 1.20 × 106 0.67
2.0 9.32 0.00179 9.94 × 10-6 9.94 × 10-7 1.01 × 106 0.50
2.5 8.88 0.00171 9.49 × 10-6 9.49 × 10-7 1.05 × 106 0.40
3.0 7.92 0.00152 8.44× 10-6 8.44× 10-7 1.18 × 106 0.33
0.0000012
0.000001
Enzyme activity, V (mol/min)
0.0000008
0.0000006
0.0000004
0.0000002
0
0 0.5 1 1.5 2 2.5 3 3.5
Substrate Concentration (%)
1.20E+06
y = -137657x + 1E+06
R² = 0.5439
1.00E+06
8.00E+05
1/V
6.00E+05
4.00E+05
2.00E+05
0.00E+00
0 0.5 1 1.5 2 2.5
1/S
Y = 0.0052x
Where:
X = protein concentration
Y= Absorbance reading
0.00168 𝑔/𝑚𝐿
Moles of glucose released (mol) = × 1 mL
180.1559 g/mol
𝑉𝑚𝑎𝑥 [𝑆]
V=
𝐾𝑚+𝑆
Double reciprocal:
1 𝐾𝑚 1 1
= +
𝑉 𝑉𝑚𝑎𝑥 𝑆 𝑉𝑚𝑎𝑥
1
y - axis =
𝑉
1
x - axis =
𝑆
1
Intercept =
𝑉𝑚𝑎𝑥
𝐾𝑚
Slope =
𝑉𝑚𝑎𝑥
1
= 1×106
𝑉𝑚𝑎𝑥
𝐾𝑚
= -137657
𝑉𝑚𝑎𝑥
Km = (-137657) × (1 × 10-6)
Km = -0.1377