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 ' while a 30% yield was obtained, both much higher than the respective values obtained using conventional methodology .