Production and over-expression of arginase with enhanced enzyme activity by an efficient recombinant Escherichia coli system

Our aim has been to develop an experimental system for the production and overexpression of arginase using genetically engineered recombinant DNA in transfected Escherichia coli cells. mRNA was isolated from the filamentous fungus, Neurospora crassa MTCC-858 (FGSC: 746{77:25C}), and cDNA encoding arginase was produced from it by RT-PCR. The cDNA was modified, ligated to a pARC035 vector, and transfected into E. coli GJ1158 and JM101 cells. Transformed colonies were selected by salt induction and IPTG-X-gal, respectively. Recombinant arginase from E.coli JM101 had 2.2 fold higher activity than the native enzyme, and was produced 3 times as quickly. In terms of both activity and economy, this recombinant arginase can be made available on a scale which could meet the high demand of its use as a potential therapeutic agent for treating a variety of cancers.