Media Optimization, Isolation and Purification of L-Asparaginase from Marine Isolate
Keywords:16S rDNA, Bacillus pumilus, enzyme purification, glutaminase side activity, L-asparaginase, marineecosystem
L-Asparaginase is an enzyme based drug used for the treatment of Acute Lymphoblastic Leukemia (ALL). In the current study, a bacterial strain was isolated from marine sediment, and screened for L-asparaginase production. On the basis of biochemical tests and 16S rDNA sequencing the organism was identified as Bacillus pumilus. The nutritional parameters affecting the L-asparaginase production by the strain were optimized. Maximum yield of Lasparaginase (157.03 ± 2.81 IU/ml) was obtained after 48h of incubation of medium supplemented with Galactose (2%) and asparagine (0.1%) as carbon and nitrogen sources, respectively. The enzyme extracted from Bacillus pumilus was partially purified by ammonium sulphate fractionation (70%), and DEAE cellulose chromatography. The apparent molecular weight of the enzyme and its subunit was 136 kDa and 71 kDa, respectively. The glutaminase activity of the enzyme was found to be 3.5 times lower than its asparaginase activity.
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