GLUTAMINASE AND UREASE-FREE ANTITUMOR EFFECTIVE L-ASPARAGINASE-PRODUCING MICROORGANISMS ISOLATION

L-asparaginase (EC 3.5.1.1) is a biopharmaceutical enzyme that catalyzes the hydrolysis of Lasparagine

to L-aspartic acid and ammonia. Currently, L-asparaginase is used clinically, for the

chemotherapeutic treatment of childhood acute lymphoblastic leukemia (ALL) and

lymphoblastic lymphoma. Recently, there had been preclinical studies showing that the antitumor

activity of L-asparaginase actually covers a wider range of cancer types (i.e., ovarian

cancer, breast cancer and glioma). L-Asparaginase also reduces acrylamide formation in food by

selectively hydrolysing asparagine to aspartic acid and ammonia without affecting other amino

acids, retaining food quality. L-Asparaginase is widely present in plants, animals and microbes

but not in humans. Microorganisms are a better source for the production of enzyme as they are

easy to cultivate and manipulate. For successful clinical applications, it is necessary to identify

glutaminase-free L-asparaginase, which is obtained from a novel producer organism. The aims

of the current study were isolation and identification of glutaminase and urease free Lasparaginase

producing novel bacteria from the digestive systems of some animals and soil. The

selection of L-asparaginase, L-glutaminase and urease producing strains was carried out by the

private plate assay containing phenol red (0.009%) as an indicator. L-asparaginase-producing

isolates that do not produce glutaminase and urease were identified by 16s rRNA sequencing of

Bacillus atrophaeus strain AspK1, Bacillus atrophaeus strain AspK3 and Brevibacterium

frigoritolerance strain AspS1.