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. 2022 Oct 21;9:1011981. doi: 10.3389/fmolb.2022.1011981

FIGURE 1.

FIGURE 1

Comparative growth assays were conducted in 96-well format between B. subtilis strain JEBS102 (Em r Cm r trpC2 ΔfadN lacA::P xyl -dcas9 amyE::P veg -sgRNA (fabF)), and the original strain BKE32840 (trpC2 ΔfadN::erm). (A) Cell growth was compared between the two strains in M9-Gluc, 5 mM L-tryptophan, 1% (w/v) D-xylose, plus the addition of 0, 5, 10, and 20 μg/ml of cerulenin. In the absence of cerulenin, BKE32840 displayed a shorter lag time but ca. 2-Fold lower cell densities that JEBS102. A cerulenin concentration of 5 μg/ml did not inhibit growth of BKE32840 but significantly inhibited JEBS102 with low turbidity developing in some wells after 37 h of incubation. Higher concentrations of cerulenin blocked growth of both strains. (B) B. subtilis strains JEBS102 and BKE32840 were grown in M9-Gluc (with 5 mM L-tryptophan), 1% (w/v) D-xylose, 20 μg/ml cerulenin, 5 g/L FA-free BSA and either with or without the addition of 8 μg/ml each of n16:0 and a15:0. The addition of FAs rescued the growth of both strains with JEBS102 displaying a slower rate of growth but higher overall cell densities. No growth occurred in the absence of exogenous FAs. Each data point represents the mean of three biological replicates with error reported as ± standard deviation.