Cloning and optimizing the expression of asparaginase enzyme from Bacillus subtilis

BACKGROUND AND OBJECTIVESL-Asparaginases (L-ASNase, EC ۳.۵.۱.۱) are enzymes that catalyze the hydrolysis of L- asparagine to L-aspartic acid and are found in various organisms from microorganisms to mammals. However, they are mainly expressed and produced by microorganisms. Microbial L-asparaginases have received ongoing attention because of their unique role in the treatment of acute lymphoblastic leukemia and because they inhibited acrylamide formation during food processing.MATERIALS AND METHODSThe asparaginase enzyme gene was amplified from Bacillus bacteria and placed in the pet۲۶ vector by heat shock method and transformed into BL۲۱ bacteria and its expression was induced by EPTG.RESULTS AND DISCUSSIONTo confirm the methods of bacterial growth in two culture mediums containing the drug kanamycin and without the presence of this drug, it was observed that the transformed bacteria grew in the culture medium containing kanamycin and this method was confirmed by PCR and electrophoresis techniques.CONCLUSIONSince the asparaginase enzyme was produced well in this part of the research, it can be used to advance therapeutic goals in other aspects of research

کلیدواژه ها:

Cloning ، Expression ، Enzyme ، Asparaginase