rLysozyme Solution: Stabilized Recombinant Lysozyme

The rLysozyme Solution contains a highly purified and stabilized recombinant lysozyme that can be used for lysis of E. coli. The enzyme catalyzes the hydrolysis of N-acetylmuramide linkages in bacterial cell walls. The specific activity ofrLysozyme solution (1,700 KU/mg) for E. coli lysis is 250 times greater than that of chicken egg white lysozyme. The rLysozyme solution is optimally active at physiological pH. Small amounts of rLysozymesolution (3–5,000 U/gram cell paste) enhance the efficiency of protein extraction with BugBuster®, BugBuster® HT and PopCulture Reagents. In the absence of protein extraction reagents, direct lysis of E. coli can be achieved by treatment of 1.0 gram cell paste with 45–60 KU rLysozyme solution, which is supplied ready-to-use at 30 KU/µL in 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton X-100. The rLysozyme solution is stable at –20 °C.

Unit Definition: One unit of rLysozyme solution is defined as the amount of enzyme necessary to cause a decrease of 0.025 A450 units per minute at 25 °C in a 1.0 mL suspension (1 mg/mL) of Tuner(DE3) cells in 0.5 × BugBuster® reagent diluted with 50 mM Tris-HCl, pH 7.5.

 

Comparison of chicken egg white lysozyme and rLysozyme activities

Comparison of chicken egg white lysozyme and rLysozyme solution activities. Activities were measured in a standard activity assay.

Materials