ISOLATION OF CHITIN DEACETYLASE FROM MUCOR-ROUXII BY IMMUNOAFFINITY CHROMATOGRAPHY

The purification of chitin deacetylase from Mucor rouxii to homogeneit y employing conventional methods has already been described. However, a lengthy protocol is required resulting in a low yield and specific a ctivity for the enzyme. A 169-fold one-step purification of chitin dea cetylase by immunoaffinity chromatography is reported, resulting in a homogeneous enzyme preparation. The enzyme purified using this procedu re was judged to be electrophoretically homogeneous as tested by both native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl su lphate PAGE. Using antibodies of lower affinity, less severe chemical conditions were required for the desorption of immunoadsorbents. Chiti n deacetylase purified by immunoaffinity chromatography exhibited a sp ecific activity of 13 U mg-1 while a 30% yield was obtained, both much higher than the respective values obtained using conventional methodo logy.