Fig. 3
Selected hits from the lysate activity screens of E. coli transformant libraries generated by site-saturation mutagenesis. Potential hits for the F336 and Q334 libraries were selected based on two individual criteria: (I) high KIC consumption and (II) altered consumption for one or both substrates. A. Screening result for the selected colonies from the Q334X mutant library. Relative consumption rate of KIC (green) and HPP (blue) by Q334X mutants in comparison to WT with the introduced codon and translated amino acid below. Relative consumption rate with respect to the WT (set to 1.0) was calculated by normalizing the consmuption over time based on OD600. B. Screening result for the selected colonies from the F336X mutant library. Relative consumption rate of HPP (blue) and KIC (light green) by F336X mutants in comparison to WT with the introduced mutation below. WT and empty vector control (EVC) for comparison are shown as average with a standard deviation from eight wells per 96-well plate