Cloning and expression of l-Asparaginase gene from Aspergillus terreus in E.coli

In the present study, L-Asparaginase was isolated from fungal source (Aspergillus terreus) and it is used as the treatment of malignancies. The synthesis of L-asparaginase from two mutant strains, WF and 933 of Serratia marcescens is reported and different media composition alters the levels of enzyme production. The phenotype was conducted by using a light microscope. While the genotype of the strain was also identified. The 18S rDNA sequences of the strain were determined by PCR. The enzyme was purified using ammonium sulphate precipitation, Sephadex G100, CM-cellulose and DEAE Sephadex chromatography.. The molecular weight of the l-Asparaginase from E.coli was determined by SDS-PAGE gel electrophoresis and it was found to be a single protein band with approximately 43 kDa.

Keywords:

cloning , PCR , component , L-Asparaginase