RAPID DETECTION OF A FUNGAL POLY (3-HYDROXYBUTYRATE) DEPOLYMERASE USING A NEW IN-SITU GEL ACTIVITY STAINING TECHNIQUE

A modified and rapid technique has been developed for activity stainin g of a fungal poly(3-hydroxybutyrate) depolymerase by incorporating po ly(3-hydroxybutyrate) (2.5% w/v) in the polyacrylamide gel under polym erisation and incubating in 100 mM Tris/HCl (pH 8.0) containing 1 mM C aCl2 at 30 degrees C. This decreased the time required for detection t o 4 h from the conventional 12-18 h using solid-plate method. The enzy me preparation failed to penetrate the gel on conventional electrophor esis but treatment with 0.5% (v/v) Triton X-100 facilitated enzyme mig ration into the gel without its activity being affected.