Nitrile hydratase of Brevibacterium R312--purification and characterization

Nitrile hydratase was purified and crystallized from the crude extract of Brevibacterium R312 and found to be homogeneous by the results of disc gel electrophoresis, analytical ultracentrifuge and double diffusion in agarose. The enzyme has a molecular mass of about 85,000 Da and contains approximately 3 g atoms iron/mol enzyme. The enzyme was composed of two kinds of subunits, of which molecular masses were 26,000 Da and 27,500 Da. The concentrated solution of the enzyme had a pronounced greyish green color and exhibited a broad absorption in visible range with a absorption maxima at 712 nm. The enzyme was active toward various aliphatic nitriles.