Purification of L-asparaginase from a bacteria Erwinia carotovora and effect of a dihydropyrimidine derivative on some of its kinetic parameters.
L-Asparaginase shows antileukemic activity and is generally administered in the body in combination with other anticancer drugs like pyrimidine derivatives. In the present study, L-asparaginase was purified from a bacteria Erwinia carotovora and the effect of a dihydropyrimidine derivative (1-amino-6-methyl-4-phenyl-2-thioxo, 1,2,3,4-tetrahydropyrimidine-5-carboxylic acid methyl ester) was studied on the kinetic parameters Km and Vmax of the enzyme using L-asparagine as substrate. The enzyme had optimum activity at pH 8.6 and temperature 35 degrees C, both in the absence and presence of pyrimidine derivative and substrate saturation concentration at 6 mg/ml. For the enzymatic reaction in the absence and presence (1 to 3 mg/ml) of dihydropyrimidine derivative, Km values were 7.14, 5.26, 4.0, and 5.22 M, and Vmax values were 0.05, 0.035, 0.027 and 0.021 mg/ml/min, respectively. The kinetic values suggested that activity of enzyme was enhanced in the presence of dihydropyrimidine derivative.
Kamble
V P
VP
School of Life Sciences, Biotechnology Research Laboratory, Swami Ramanand Teerth Marathwada University, Nanded (MS) 431 606.
Rao
R Srinivasa
RS
Borkar
Prita S
PS
Khobragade
C N
CN
Dawane
B S
BS
D001215:Asparaginase / Q000302:isolation & purification* / Q000502:physiology*; D007700:Kinetics; D016973:Pectobacterium carotovorum / Q000201:enzymology*; D011743:Pyrimidines / Q000494:pharmacology*
D011743:Pyrimidines; D001215:Asparaginase