Journal: Communications Biology

Article Title: An ancient lysozyme in placozoans participates in acidic extracellular digestion

doi: 10.1038/s42003-025-09409-6

Figure Lengend Snippet: A SDS-PAGE gel of purified recombinant mPLys_M112 and pPLys. The molecular mass standard is indicated on the left and marked with the associated molecular masses in kDa. B Lysozyme assay of 32 pmol pPLys, 32 pmol mPLys_M112, and 32 pmol HEWL at pH 5.0. Reaction buffer consisted of 20 mM sodium acetate, pH 5.0 (I = 13 mM). Dependency of lysozyme activity of pPLys and mPLys_M112 on ionic strength ( C ) and pH ( D ) at room temperature. E Lysozyme activity of pPLys, mPLys_M112 and HEWL at different temperatures. All measurements were performed in triplicates. Bacterial lysis assay against B. subtilis ( F ) and E. coli D31 ( G ) using a membrane-impermeable dye (SYTOX Green). Fluorescence indicates lysis of cells. Sigmoidal functions were fitted to the dose response curves. mPLys_M112: open triangles, pPLys closed triangles, HEWL closed squares, Melittin closed circles, buffer control: crosses.

Article Snippet: 1 × 10 6 CFU/well E. coli K12 D31 and 1 × 10 5 CFU/well in mid-logarithmic growth phase were added to a dilution series of the protein of interest in a black flat-bottom 96-well plate (Sarstedt), together with the DNA binding dye SYTOX Green (Invitrogen).

Techniques: SDS Page, Purification, Recombinant, Activity Assay, Lysis, Membrane, Fluorescence, Control