Extraction, Purification and Characterization (A) of Arginine Deiminase Enzyme from a Local Higher Productive Isolation of Enterococcus faecium M1

Abstract

Arginine deiminase (ADI) enzyme was extracted from Enterococcus faecium M1 isolate and lysozyme was efficient in bacterial cell lyses more than Triton-X100 and Glass beads.ADI was more active in the range of (50-80%) ammonium sulfate saturations. After purification by anion exchange and gel filtration chromatography the specific activity of ADI reached to 59.2 U/mg protein with 11.23 folds and 42.93% yield. The purity of enzyme was estimated by Native-PAGE electrophoresis under non denatured conditions. The whole average molecular weight of ADI was 186 KDa, included two non- identical bands on SDS- PAGE, one of them had 52 KDa and the other had40 KDa. By concluding, ADI may contain four polypeptide subunits (tetrarmeric enzyme). Optimum pH for enzyme activity was ranged between (6.5 to 7.5) with maximum activity at pH 7.0 and still active over a wide range of pH values (4.0-10), ADI was more stable at pH level (6.0-7.5) with full remaining activity. By concluding, the maximum activity and stability of ADI in neutral pH encouraged us to use it as anticancer treatment agent and for other applications in human body.

Keywords: Arginine deiminase, E. faecium M1, Extraction, Purification, Characterization (A).