Does the concentration of these enzymes affect their ability to reduce the visco
ID: 274115 • Letter: D
Question
Does the concentration of these enzymes affect their ability to reduce the viscosity of the glucan solutions? Why?
y = 0.0294x + 0.1145
R2 = 0.994
Y
X
Water
Protein
Absorbance (Abs)@595nm(duplicates)
Absorbance (Abs)@595nm(duplicates)
Mean Abs
the amount of BSA
total Volume
dilution factor
Final Protein (enzyme) concentration (?g/ml)
Final Protein (enzyme) concentration (mg/ml)
Dilution 1 :?-glucanase
450
50
0.386
0.43
0.408
9.983
0.5ml
10
199.66
0.19966
Dilution 2 :?-glucanase
480
20
0.225
0.229
0.227
3.827
0.5ml
25
191.35
0.19135
Dilution 1 :?-xylanase
450
50
/
0.919
0.919
27.364
0.5ml
10
547.28
0.54728
Dilution 2 :?-xylanase
480
20
0.591
0.688
0.6395
17.857
0.5ml
25
892.85
0.89285
Sample
Average time ( ?-glucanase treated)
Average time ( untreated)
Buffer time
Viscosity
Reduction Viscosity
Barley
21.42
174.397
16.082
3.37%
96.63%
Oat
17.157
58
16.082
2.56%
97.44%
Rye
60.266
68.333
16.082
84.56%
15.44%
Sample
Average time ( ?-xylanase treated)
Average time ( untreated)
Buffer time
Viscosity
Reduction Viscosity
Barley
119
174.397
16.082
65.01%
34.99%
Oat
50.24
58
16.082
81.49%
18.51%
Rye
24.977
68.333
16.082
17.02%
82.98%
y = 0.0294x + 0.1145
R2 = 0.994
Y
X
Water
Protein
Absorbance (Abs)@595nm(duplicates)
Absorbance (Abs)@595nm(duplicates)
Mean Abs
the amount of BSA
total Volume
dilution factor
Final Protein (enzyme) concentration (?g/ml)
Final Protein (enzyme) concentration (mg/ml)
Dilution 1 :?-glucanase
450
50
0.386
0.43
0.408
9.983
0.5ml
10
199.66
0.19966
Dilution 2 :?-glucanase
480
20
0.225
0.229
0.227
3.827
0.5ml
25
191.35
0.19135
Dilution 1 :?-xylanase
450
50
/
0.919
0.919
27.364
0.5ml
10
547.28
0.54728
Dilution 2 :?-xylanase
480
20
0.591
0.688
0.6395
17.857
0.5ml
25
892.85
0.89285
Explanation / Answer
Changes in the enzyme and substrate concentrations affect the rate of reaction of an enzyme-catalyzed reaction. Generally increase in substrate concentration increases the rate of reaction. This is because more substrate molecules will be colliding with enzyme molecules, so more product will be formed. Changing the concentration of a substance affects the rate of reaction if it is the limiting factor. Increase in enzyme concentration generally increases will the rate of reaction .This is because more enzymes will be colliding with substrate molecules. After the reaction, release of product from the active site is the rate-limiting step. The release of product is associated with enzymatic conformational change. Solvent characteristics such as viscosity strongly influence protein conformational changes. Viscosity results in friction against proteins and causes decrease in motion inhibiting catalysis in enzymes. A direct correlation was found between viscosity (?) and the inhibition of the maximum rate of catalysis (V max). The overall activity of an enzyme decrease at the saturation level of substrate concentration when the solvent viscosity is increased. The transfer of the reaction intermediate from one active site to another is required for the catalytic reaction.
In the present study, the concentration of the glucanase did not vary significantly among the two dilutions. In case of second enzyme xylanase, the concentration varies with dilution. If enzyme concentration is less than the viscosity reduction is more. The viscosity is also less in these cases. In contrast, the viscosity reduction is less if the enzyme concentration is more. The viscosity is also more in these cases. In case of enzyme xylanase similar trends has been noted. If the enzyme concentration is less, viscosity reduction is more and if the enzyme concentration is more, viscosity reduction is less.
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