[Purification and characterization of streptococcal hyaluronate lyase (author's transl)]

Hyaluronate lyase of group C strain H 46A (Streptococcus equisimilis) was purified and characterized by isoelectric focusing, sodium-dodecylsulfate-acrylamide electrophoresis, polyacrylamide-gradient-electrophoresis and crossed immunoelectrophoresis. The purification of the hyaluronate lyase was performed successively by adsorption on Florisil, chromatography on DEAE-cellulose, and - for separation of streptokinase - stirring with CPG-pore-glass. The last step was Sepharose 6B. PUrified hyaluronate lyase showed a high specific activity. The purified enzyme was found to be antigenically homogeneous. No contaminating streptococcal components could be detected. The molecular size was determined as to be 90 000 dalton by gel filtration and 110 000 dalton by sodium dodecylsulfate-acrylamide electrophoresis. The amino acid composition was also determined. In the isoelectric focusing, using gels with reducing conditions, one protein band at a pI 4.95 was observed. Under nonreducing conditions two or three diffuse protein bands which showed lower enzymatic activity were found. It might be possible that the hyaluronate lyase exists in two different forms.