Purification of Arginine Deiminase From Pseudomonas alcaligenes

Key words: 
• arginine deiminase /
• purification /
• ammonium sulfate graded precipitation /
• column chromatography

Abstract: Crude arginine deiminase（ ADI） enzyme was extracted from Pseudomonas alcaligenes,purified by using ammonium sulfate graded precipitation,and then further separated and purified by using DEAE-Sephadex A-50 and Sephadex G-100 column chromatography to produce purified ADI. The purity of the purified ADI was determined by SDS-PAGE electrophoresis. The total enzyme activity of the ADI crude enzyme was detected to be 424. 43 U with a specific enzyme activity of 0. 42 U; the enzyme purified via salting out and dialysis had a total activity of 222. 10 U with a specific enzyme activity of 2. 05 U·mg^-1; the enzyme purified by ion exchange analysis showed a total activity of 105. 83 U at the active site and its specific enzyme activity was 5. 61 U·mg^-1; the enzyme purified through gel permeation chromatography had a total activity of 17. 94 U at the active site and its specific enzyme activity was 11. 80 U·mg^-1. The purified ADI had a relative purity of 96%,which can be used for electrophoresis. It is concluded that this method can be used to produce ADI with higher purity and activity.