Cloning and Recombinant Expression of L-asparaginase from Bacillus cereus

L-Asparaginase plays a vital role as a therapeutic enzyme in the treatment of acute lymphoblastic leukemia (۱). The L-asparaginase enzyme catalyzes the conversion of the amino acid L-asparagine to L-aspartic in addition to ammonia (۲). This enzyme has been widely applied in the pharmaceutical and food industries (۳). Asparaginase is distributed widely among microorganisms, plants and animals, microorganisms are considered an excellent source for producing the L-asparaginase due to the convenience for large-scale production and purification (۴). In this reaserch ,the asparaginase gene was isolated, cloned and expressed from Bacillus cereus. For this purpose, the relevent gene was first isolated by PCR and then cloned in pET۱۹b vector with appropriate restriction enzymes, and then expressed in the E.coli BL۲۱ expression system.The optimal expression of the soluble enzyme was expressed at ۲۵ ℃ for ۴ hours. The analysis of the expression results on ۱۲% SDS-PAGE gel confirmed the presence of a protein with an approximate molecular weight of ۳۵ kDa. The high expression rate of the recombinant enzyme and the appropriate ratio of its soluble to insoluble expression indicates the potential of this enzyme for further biochemical studies and its high-volume production with application in various biological industries

کلیدواژه ها:

Bacillus cereus ، L-Asparaginase ، Recombinant expression ، pET۱۹b