Lactic Streptococcal Phage-Associated Lysin. II. Purification and Characterizaton

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Abstract
C10-Lysin was purified approximately 1,700-fold by a multistep procedure involving use of the ultracentrifuge, ammonium sulfate fractionation, gel filtration, and ion exchange chromatography. The purified lysin was very labile. The presence of 2-mercaptoethanol and EDTA preserved but did not increase c10-lysin activity. Both mono-and divalent cations activated c10-lysin. c10-Lysin activity was inhibited by sulfhydryl binding agents. The purified c10-lysin revealed a pH optimum of 6. 0-6. 5 Q10 and energy of activation values of 3. 9 and 25 Kcal, respectively, and migrated electrophoretically toward the cathode at pH 8. 6. The clO-lysin failed to show proteolytic activity. Crude preparations of another lactic streptococcal phage-associated lysin (c2-lysin) showed physico-chemical characteristics similar to crude and purified clO-lysin prepartions.