Fig. 3: Generation of salivaricin B mutants and their antimicrobial activity.
From: Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides
a Representative MALDI-TOF-MS analyses of single-site mutagenesis at the position of Q6 or L20. b Representative MALDI-TOF-MS analyses of two-site mutagenesis at the positions of Q6 and L20. In (a) and (b), salivaricin B-1 based variants are the most abundant products in all analyses, which are indicated with the [M + H]+ ions. c Quantification of salivaricin B mutants that are synthesized in UniBioCat reactions (see Supplementary Table 5 for the yield of each peptide variant). d Antimicrobial activity assay showing ΔOD600 profiles of S. aureus RN4220 growth treated with single-site (top) and two-site (bottom) mutants. Note that cell-free reaction mixture is used directly for the assay without purification of each mutant. NC, negative control without gene templates in the cell-free reaction; Kan, kanamycin (25 μg/mL) as a positive control. Data shown representative of three independent experiments (n = 3). In (d), data are presented as mean±s.d. of two independent experiments (n = 2). Source data are provided as a Source Data file.
