(A) In vitro competition assay with 1 mM of 5-ASA,
4-ASA, procainamide, hydralazine and isoniazide demonstrates relative
specificity of the Firmicutes CAG:176 thiolase
(FcTHL) for 5-ASA. N=3 biologically independent samples
per enzyme/condition. (B) Shown is a representative Michaelis-Menten plot of
n=1 thiolase enzyme preparation, conducted in technical triplicates at each
concentration of 5-ASA; summary data is from n=5 biologically independent
experiments each conducted in technical triplicate. (C) Live culture of
Oscillibacter sp., strain KLE 1745 encoding another predicted thiolase gene
(UniRef90 R6TIX3) was capable of acetylation of 5-ASA to
N-acetyl 5-ASA. N=3 biologically independent samples per
condition, representative data from n=2 independent experiments, unpaired,
two-sided T-test, *** = p = 0.0015. Data are presented as mean values
+/− SEM.