Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Basedon HaloTag

Data underlying the figures in the publication “Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on Halotag” published in ACS Catal., 2021, 11, 6343−6347. The data contains an excel file with the curate data of the graphs from figures 2, 3 and 4. Data for Figure 2: Data processing: the data collection began from SDS gels (a representative gel is shown in the Supporting Information and at the bottom of this table). The total fluorescence of HaloTag was measured using the fluorescence-compatible gel imager. The signal in each lane was quantified with ImageJ. The signal was processed as indicated in the supporting information. Data for Figure 3: Data processing for fluorescence measurements: the data processing began with fluorescence intensities measured using a fluorescence plate reader. A calibration curve containing varying product concentrations was present on every plate. A linear fit of the calibration curve (fluorescence intensity vs. product concentration) was used to calculate the concentration of product in each experimental well. A sample of the calibration curves is supplied in the supporting information. The turnover number (TON) of each well was determined by dividing the product concentration by the concentration of the catalyst. Data processing for MS measurements: the data processing began with chromatogram from the GC-MS or the UPLC-MS. The peak areas corresponding to the desired product were integrated. A calibration curve containing varying product concentrations was completed for each desired product. A linear fit of the calibration curve (peak area vs. product concentration) was used to calculate the concentration of product in each reaction. A sample of the calibration curves is supplied in the supporting information. The turnover number (TON) of each well was determined by dividing the product concentration by the concentration of the catalyst. Data for Figure 4: Data processing for MS measurements: the data processing began with chromatogram from the UPLC-MS. The peak areas corresponding to the desired product were integrated. A calibration curve containing varying product concentrations was completed for each desired product. A linear fit of the calibration curve (peak area vs. product concentration) was used to calculate the concentration of product in each reaction. A sample of the calibration curves is supplied in the supporting information. The turnover number (TON) of each well was determined by dividing the product concentration by the concentration of the catalyst. The TON relative to wild-type enzyme was determined by dividing the TON of each variant by the TON of wild-type HT with the corresponding cofactor (Mes8 or N8)