A zymographic assay for detection of hyaluronidase activity on polyacrylamide gels and its application to enzymatic activity found in bacteria.

A zymographic assay for the determination of hyaluronidase activity in cell-free extracts on native polyacrylamide gels has been developed. In this assay an agarose replica of the polyacrylamide gel which contains hyaluronic acid and bovine serum albumin (BSA) was used. After an incubation at 37 degrees C to allow transfer and development of enzymatic activity, the hyaluronic acid and BSA were precipitated in the agarose gel with 2 M acetic acid. Areas of enzymatic activity appeared as clear zones in the agarose replica. The assay was sensitive and was used to demonstrate hyaluronidase activity in cell-free extracts from a number of bacterial and mammalian species.