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FabI was preincubated with or without equimolar concentrations of triclosan and 2 mM NAD+ for 30 min in Tris/HCl (pH 7.4) at 25 °C. The preincubated sample was then diluted 200-fold into a solution of competing NADH and crotonoyl-CoA and the dissociation of triclosan was monitored by following the enzyme activity.
