anti clc 2  (Alomone Labs)

Journal: mBio

Article Title: Matrix-independent screening of defluorination in vitro and in vivo

doi: 10.1128/mbio.01798-25

Figure Lengend Snippet: Measuring substrate defluorination in vivo by pH change using a recombinant Pseudomonas expressing a defluorinase, DEF1. The reactions and cell growth were monitored by changes in bromothymol blue absorbance at 615 nm and turbidity at 700 nm (OD 700 ), respectively. The blue curves show the change in pH as a drop in absorbance at 615 nm when cells are present. The magenta curve shows absorbance at 615 nm with substrate alone, which controlled for the dissolution of carbon dioxide from the atmosphere, which gives background acidification. The orange curve shows the increase in OD 700 due to cell growth. ( A ) Reactions and growth with α-fluorophenylacetic acid. ( B ) Reactions and growth with 2-fluoropropionic acid. ( C ) Comparison of the rates of defluorination measured in a previous study with purified enzyme in vitro with the in vivo rate determined here via acidification by the change in absorbance at 615 nm. The rate was determined from the linear regions of the blue curves. The background absorbance change (magenta curve) was subtracted to correct for the control pH change. The rates were multiplied by 10 to make the numeric comparison easier. Error bars shown in parts A and B represent standard deviations from triplicate determinations.

Article Snippet: The P. putida ATCC 12633 DEF1 + CLC F psy strain was grown overnight to the stationary phase in LB with antibiotics as described above.

Techniques: In Vivo, Recombinant, Expressing, Dissolution, Comparison, Purification, In Vitro, Control