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INTRODUCTION
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Int. J. Pharm. Sci. Rev. Res., 23(2), Nov Dec 2013; n 35, 198-202
Dialysis
Kinetic Characterization
Calculation of Km and Vmax values
The Km and Vmax values of L-asparaginase were calculated
from the reciprocal plots of substrate concentration
versus reaction velocity. The Lineweaver-Burk plot (LB
plot) was constructed and it was found to be linear,
suggesting a simple Michealis-Menten (MM plot) kinetics.
199
Int. J. Pharm. Sci. Rev. Res., 23(2), Nov Dec 2013; n 35, 198-202
Total
protein(mg)
Specific activity
(IU/mg)
Purification
fold
Yield (%)
Fermented broth
114.82
6.8
16.88
----
100
92.45
0.1
924.5
54.76
80.51
Dialysis
83.41
0.08
1042.6
61.76
72.64
69.0
0.05
1380
81.75
60.09
L-asparaginase activity
(IU/mL)
0.01
1.98
0.02
3.46
0.03
4.42
0.04
5.12
0.05
5.56
0.06
5.92
0.07
5.96
0.08
5.96
0.09
5.96
10
1.25
20
2.51
30
3.773
40
5.75
50
5.52
60
4.99
70
1.2
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Int. J. Pharm. Sci. Rev. Res., 23(2), Nov Dec 2013; n 35, 198-202
Table 4: Effect of pH on L-asparaginase activity
pH
4.5
1.86
5.0
2.89
5.5
3.8
6.0
4.46
6.5
5.68
7.0
6.12
7.5
6.45
8.0
5.89
8.5
5.36
9.0
4.53
9.5
3.31
10.0
2.02
LB plot
Km
0.03M
0.03M
Vmax
5.96 IU/mL*min
9.69 IU/mL*min
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Int. J. Pharm. Sci. Rev. Res., 23(2), Nov Dec 2013; n 35, 198-202
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Kamala Kumari PV, Girija Sankar G, Prabhakar T, Lasparaginase production and molecular identification of
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202