Fig. 1
In silico and in vitro analysis of Ar-BVMO reaction with three different alkyl levulinates: methyl (R = CH3), ethyl (R = CH2CH3) and butyl (R = CH2CH2CH2CH3) levulinate. A Reaction scheme. Ar-BVMO structure was predicted with AlphaFold2 Monomer v2.0 (https://alphafold.ebi.ac.uk/entry/D7PBN2). B Absorbance at 520 nm measured after performing a high-throughput colorimetric assay for detection of esters produced by Ar-BVMO reactions with alkyl levulinates. Data points are mean ± SD of three independent experiments. Binding and interactions (black dashed lines) within Ar-BVMO’s active-site were calculated using VINA software embedded in YASARA (https://www.yasara.org); C Methyl levulinate, D Ethyl levulinate and, E Butyl levulinate. The carbonyl group involved in the BV reaction is circled in dark green. F Distances (in Å) and interaction types of each substrate with Ar-BVMO’s active site residues, cofactor (FAD) and coenzyme (NADP+). Figures C, D and E were produced by the 3D molecular visualisation software PyMOL (https://www.pymol.org/).
