Isolation and Characterization of L- Asparaginase Producing Endophytic Bacteria from Simarouba Gluaca

Abstract

Cancer cells differentiate themselves from normal cells in diminished expression of L-asparaginase. It is the enzyme that catalyzes the hydrolysis of L-Asparagine to L-aspartic and ammonia, because of these it is used as a medication and in food manufacturing. As a medication L-aspraginase is used to treat various types of leukemia such as, acute lymphoblastic leukemia, acute myeloma leukemia and non Hodgkin’s lymphoma. Hence they are not capable of producing L-asparaginase and mainly depend on the L-asparagine from circulating plasma pools. The clinical action of this enzyme is attributed to the reduction of L-asparaginase, since tumor cells unable to synthesis these amino acids are selectively killed by L-asparaginase depravation. This enzyme is widely distributed, being found in as it is widely distributed bacteria as well as found in animals, microbes and plant sources. In the present study L-asparaginase producing bacteria was isolated from Simarouba glauca . It was grown on the modified M9 medium in which L- asparagine was the major source of L- asparaginase production was detected by the formation of pink colored zones on the medium. After the partial purification of the L-Asparaginase enzyme, the enzyme activity was found to be 155.83 Units/ml and specific activity of 779.15. The optimum pH was to be found at pH 8 at a temperature of 37°C in the presence of 10mM Mg 2+ . The molecular identification was done by16S rDNA, PCR and sequence analysis by BLAST further confirmed that Bacillus cereus .