P91: Cloning and Expression of three L-Asparaginases of Erwiniacarotovora subsp. atroseptica in Escherichia coli

L-Asparaginase (EC 3.5.1.1), which catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia, has been used as therapeutic agent for the treatment of acute lymphoblastic leukemia and lymphosarcoma. The subcellular distribution of L-asparaginase in Erwinia carotovora subsp. atroseptica confirmed that the presence of cytoplasmic, periplasmic and membrane bound enzyme and maximum was located in the cytoplasm. There are three different genes for L-asparaginase (L-assparaginase,L-asparaginase-I and L-asparaginase-II) were found in E. carotovora subsp. atroseptica.  In order to study the individual gene expression, we have successfully cloned and expressed all three genes individually in E.coli (BL21DE3). The expression levels of all three genes are much higher as compared to L-asparaginases from Erwinia carotovora subsp. atroseptica.