Fig. 2: Evolution of AtaPT for efficient synthesis of prenylated kempferol derivatives. | Nature Communications

Fig. 2: Evolution of AtaPT for efficient synthesis of prenylated kempferol derivatives.

From: Tuning aromatic cage occupancy in prenyltransferases enables selective and efficient production of rare c-prenylated flavonoids

Fig. 2: Evolution of AtaPT for efficient synthesis of prenylated kempferol derivatives.

a Reactions starting with KAE to form 8P-KAE and Icaritin. b HPLC chromatogram showing the conversion of 1a with DMAPP to products 2a, 3a, 4a, and 5a catalyzed by AtaPT-WT. c HPLC chromatogram of 1a with GPP resulting in the formation of product 6a catalyzed by AtaPT-WT. d Improvement in catalytic activity across five rounds of directed evolution using site-saturation mutagenesis (SSM), iterative saturation mutagenesis (ISM), and focused rational iterative site mutagenesis (FRISM). WT: wild type, M2: S177G/P324H, M3: S177G/P324H/N328M, M5: E169A/S170Q/S177A/P324H/N328M, M7: I167T/E169A/S170Q/S177G-/L323Q/P324H/N328M and M8: I167A/E169A/S170Q/C175M/S177A/L323Q/P324H/N328R. e Rescreening of mutants using GPP to enhance the conversion efficiency of (6a). f HPLC chromatogram showing the conversion of 1a with DMAPP to products 2a, catalyzed by AtaPT-M8. g HPLC chromatogram of 1a with GPP resulting in the formation of product 6a catalyzed by AtaPT-M7. Relative activities are reported as mean ± SD of three independent biological replicates (n = 3). The yield was determined by substituting the peak area into the standard calibration curve.

Back to article page