Extended Data Figure 5. Characterization of selected proteins identified in the supernatant of Leaf245.

a) List of proteins identified in the supernatant of Leaf245 and selected for heterologous expression in E. coli. For each gel band in Fig. 3d as well as the most active fractions from two independent experiments the rank abundance of the proteins based on total ion count as identified by mass spectrometry are shown. b, c, d) Leaf374 inhibition and lysis by heterologously expressed candidate proteins. b) Activity of cleared cell lysate after expression of proteins (top label) in E. coli BL21 DE3 gold. Lysate was applied on Leaf374 overlay plates directly after preparation (left panel) or on Leaf374 overlay plates that were pre-incubated for 24 h to assess lysis (right panel). Lysate concentration was normalized to the final OD₆₀₀ that each expression culture reached. Up to 50-fold dilutions in the buffer used for lysis (see methods) were prepared as indicated on the left. c) Activity of purified proteins against 24 h pre-grown Leaf374. d) Activity of ASF05_00205 native and mutant (S70L) protein against Leaf374. Concentrations of both purified proteins were normalized (0.1 mg mL^-1) and a 10- and 100- fold dilution was prepared and assayed on a Leaf374 overlay plate directly after preparation (left panel) or after the overlay was pre-incubated for 24 h (right panel) to test for growth inhibition and cell lysis, respectively. Data shown for ASF05_00205 native protein is the same as shown in Fig. 3e and panel c) of this figure.