ISOLATION OF CHITIN DEACETYLASE FROM MUCOR-ROUXII BY IMMUNOAFFINITY CHROMATOGRAPHY

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