FIG 9.

Phenotypic analysis of EVT strains. (A) Comparison of levels of growth in LB plus 10 mM vanillin at 6 h after inoculation (OD₆₀₀) of BW25113 wild-type parent and EVT strains 1 to 4. A value of 100% corresponds to an OD₆₀₀ of 0.49 ± 0.04. (B) Specific activity of citrate synthase (GltA) in wild-type and EVT strain cell extracts (cells were grown in LB without vanillin). (C) Comparison of levels of growth in LB plus 10 mM vanillin at 6 h after inoculation (OD 600 nm) of BW25113 wild-type parent and a single gene deletion mutant in the rob gene, derived from the Keio collection. A value of 100% corresponds to an OD₆₀₀ of 0.49 ± 0.04. In panels A to C, the data represent means of results from three independent cultures or assays, with standard deviations shown as error bars. *, P = 0.01 to 0.05; **, P = 0.001 to 0.01; ****, P < 0.0001 (one-way ANOVA [A and B] or Student's t test [C]). (D) The ∼10-kb chromosome deletion in EVT4 and the effect of individual single-gene knockouts in this region on vanillin tolerance. (Upper panel) The 9,699-bp region deleted in EVT4, encompassing genes yneG to eamA with partial deletion of dcgF, as shown by the sequence read density (gray vertical bars). (Lower panel) Comparison of levels of growth in LB plus 10 mM vanillin at 6 h after inoculation (OD₆₀₀) of BW25113 wild-type parent and single-gene-deletion strains from the Keio collection. A value of 100% corresponds to an OD₆₀₀ of 0.49 ± 0.04. Red bars indicate significantly decreased growth (P < 0.05 by one-way ANOVA), and green bars significantly increased growth (P < 0.05 by one-way ANOVA) compared to the wild type. Gray bars indicate no significant change.