Lysis of S. aureus cell suspension

Staphylococcus aureus ATCC 25923 is selected as a model organism for evaluating enzyme ability to lyse bacteria. 100 µl of night culture were added to 5 ml of LB medium and cells were incubated at 37 °C, 200 rpm until OD600 reached 2. Bacteria were centrifuged and resuspended in reaction buffer (25 mM HEPES, 100 mM NaCl, pH 7.5). S. aureus culture was diluted to OD600 1 and 5 µM of enzymes were added to a total of 100 µl of reaction volume. Cells were incubated in 96-well plate at 37 °C and OD600 was measured every 5 minutes.