Molecular cloning of putative L-asparaginase/glutaminase gene from Klebseilla peunomonae MGH 78578

This study is one among a few local studies that have been reported to clone the putative L-asparaginase/glutaminase produced by recombinant gene based on the use of plasmid Ecoli - Nterm 6X HN(PLEICS-01) vector. In this study, a pair of primers was designed to obtain L- glutamin-(asparagine)ase encoding gene from Klebsiella pneumoniae subsp. pneumoniae MGH 78578. The putative L-asparaginase/glutaminase gene encoding gene (KPN-01165) was expressed in E.coli BL21(D3) to make sure the presence of a functional putative L-asparaginase/glutaminase in the cloned DNA. The recombinant L- glutamin-(asparagine)ase gene was 1005 bps which encoding a protein of 334 amino acids. The recombinant enzyme containing a 6xHistidine-tag can easily purified by Ni-NTA chromatography and its molecular weight was determined to be 40 KDa by SDS-PAGE. A putative L-asparaginase/glutaminase catalysed the hydrolysis is not only of L-glutamine, but also of L-asparagine.