Figure 4. An Acyl-CoA metabolic defense system in the Bacteroides.
(A) Tn-seq of Bt VPI grown in Glc −/+ butyrate. Specific genes are highlighted in green (BT_3942) and blue (ilv and leu operons). See also Table S3.
(B) Genomic locus of BT_3942 encoding Acyl-CoA thioesterase and predicted enzymatic reaction.
(C) Bt WT, Bt Δ3942 and the complemented strain grown in Glc −/+ butyrate.
(D) Tn-seq of Bv ATCC grown in Glc −/+ butyrate. Specific genes are highlighted in green (BVU_1164/ 1163 operon). See also Table S3.
(E) Genomic locus of the BVU_1164/1163 operon encoding a hypothetical protein and Acyl-CoA transferase and predicted enzymatic reaction.
(F) Bv WT, Bv Δ1163, and the complemented strain (two left), and Bt WT complemented with BVU_1163 (third from left). Bv Δ1163 complemented with BT_3942 (right) grown in Glc −/+ butyrate.
(G) Relative abundance of 13C butyryl-CoA in Bt WT, Bv WT, and their isogenic mutant strains 30 min post pulse at mid log phase in Glc with 13C butyrate. See also Figure S3E.
(H) Relative abundance of Acyl-CoAs in Bt WT, Bv WT, and their isogenic strains, as indicated, in Glc with butyrate or water control. See also Figure S3G.
In (A and D), plots are the combined fold change and p value from two independent replicates (See STAR Methods). Shown is a representative of three biological replicates in (C and F). Error bars represent mean ± SEM of two (G) and three (H) biological replicates. Nonsignificant p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; unpaired two-tailed t test in (G), two-way ANOVA with Tukey’s multiple comparisons test in (H).