Figure 1.
Antimicrobial overlay plate for detecting organisms from the feral hog ear that inhibited S. aureus growth.
Bacterial isolates were streaked in “X” shaped patterns onto half-strength TSA agar. The B. laterosporus PE36 isolate used in this investigation is indicated by the arrow.
Figure 2.
Phylogeny of B. laterosporous PE36 and several related B. laterosporous isolates.
Phylogeny is depicted by (A) maximum likelihood of 16S rRNA gene sequences and (B) a consensus network based on whole genome comparisons. Bootstrap values >70% are listed for nodes in the maximum likelihood tree with the scale bar representing 0.05 substitutions per position. Branch lengths for the genome consensus network are proportional to the number of likely inversion events of least collinear blocks inferred by Mauve.
Table 1.
Secondary metabolite biosynthetic gene clusters identified by antiSMASH.a
Figure 3.
Structures of secondary metabolites isolated from B. latersporus PE36.
Table 2.
1H (500 MHz) and 13C (125 MHz) NMR data (DMSO-d6, 25°C) for auripyrazine (5).
Table 3.
1H (500 MHz) and 13C (125 MHz) NMR data (DMSO-d6, 25°C) for the two predominant solution conformers of auriprocine (6).
Table 4.
MSn fragments used to support the planar structure of auriprocine (6).a
Figure 4.
Pymol representation of auriprocine (6).
Images generated from X-ray data for 6. The top two images depict the two opposing faces of the structure, rotated 180°. The bottom image is the helical wheel representation, viewed from the N-terminus along the helix axis. An animated image of 6 is provided in Movie S1.
Figure 5.
Interpretation of antiSMASH results and proposed biosynthetic origins of basiliskamides A (1) and B (2) and auriporcine (6).
Panel A illustrates the biosynthetic gene cluster hypothesized to be responsible for the production of 6. Adenylation domains are labeled with the amino acids they are predicted to contribute. The two domains potentially involved in loading of the L-leucine residues are contained in the shaded box. The additional domains shaded grey are believed not to be involved in the production of 6 or were incorporated, but later removed from the metabolite. Panel B shows the organization of the putative basiliskamide gene cluster, as well as a proposed biosynthetic scheme for the production of the polyketide portions of 1 and 2. The fused, tandem AT domains are separate from the other modules. The dehydrating bimodule is shown split, as the gene that encodes the two KS domains are separated from the dehydrogenase and ketoreductase. The genes encoding the predicted aminotransferase, condensation, and adenylation domains (hypothesized to transform the amide and add the cinnamic acid moieties to the polyketide chain) are labeled. The putative functions of the genes are indicated as A (adenylation), C (condensation), KR (ketoreductase), E (epimerization), GNAT (GCN5-related acetyltransferase), KS (ketosynthase), ER (enolreductase), MT (methyltransferase), DH (dehydrogenase), and AT (acetyltransferase).