Activities of SAG "soluble lysin"
500 ml of crude soluble lysin (green) was centrifuged 10 min at 5K. The supernatant was brought to 30% ammonium sulfate and centrifuged 10 min at 6K. The supernatant was adjusted to 65% ammonium sulfate and centrifuged 10 min at 6K. The pellet was dissolved in 25 ml of pH 6 MES buffer (25mM KOH, 50mM MES). The solution scattered light and was reasonably viscous. It was centrifuged 30 min at 30K. The supernatant ("soluble lysin") was stored at 5C.
10 microliter aliquots were added to various 4MU substates and incubated for 3 hrs at 37C along with blanks containing substrate alone.
The lysin contains substantial beta-NAcetyl- glucosaminidase, alpha-glucosidase ane beta-galactosidase activities and detectable cellobiosidase (cellulase).
In future preparations it will be advisable to give preparations a more substantial initial centrifugation to remove intact virions as well as subcellular debris.
