7YWV

Eugenol oxidase from rhodococcus jostii: mutant S81H, D151E, A423M, H434Y, S394V, Q425S, I445D, S518P

Summary for 7YWV

• Entry DOI: 10.2210/pdb7ywv/pdb
• Descriptor: Probable vanillyl-alcohol oxidase, FLAVIN-ADENINE DINUCLEOTIDE, 2-methoxy-4-(prop-2-en-1-yl)phenol, ... (5 entities in total)
• Functional Keywords: biocatalysis, thermostability, oxidase, flavoenzyme, oxidoreductase
• Biological source: Rhodococcus jostii RHA1
• Total number of polymer chains: 8
• Total formula weight: 478278.64

Authors

Alvigini, L.,Mattevi, A. (deposition date: 2022-02-14, release date: 2022-11-16, Last modification date: 2024-01-31)

Primary citation

Guo, Y.,Alvigini, L.,Trajkovic, M.,Alonso-Cotchico, L.,Monza, E.,Savino, S.,Maric, I.,Mattevi, A.,Fraaije, M.W.
Structure- and computational-aided engineering of an oxidase to produce isoeugenol from a lignin-derived compound.
Nat Commun, 13:7195-7195, 2022

PubMed Abstract: Various 4-alkylphenols can be easily obtained through reductive catalytic fractionation of lignocellulosic biomass. Selective dehydrogenation of 4-n-propylguaiacol results in the formation of isoeugenol, a valuable flavor and fragrance molecule and versatile precursor compound. Here we present the engineering of a bacterial eugenol oxidase to catalyze this reaction. Five mutations, identified from computational predictions, are first introduced to render the enzyme more thermostable. Other mutations are then added and analyzed to enhance chemoselectivity and activity. Structural insight demonstrates that the slow catalytic activity of an otherwise promising enzyme variant is due the formation of a slowly-decaying covalent substrate-flavin cofactor adduct that can be remedied by targeted residue changes. The final engineered variant comprises eight mutations, is thermostable, displays good activity and acts as a highly chemoselective 4-n-propylguaiacol oxidase. We lastly use our engineered biocatalyst in an illustrative preparative reaction at gram-scale. Our findings show that a natural enzyme can be redesigned into a tailored biocatalyst capable of valorizing lignin-based monophenols.

PubMed: 36418310
DOI: 10.1038/s41467-022-34912-3

Experimental method

X-RAY DIFFRACTION (2.4 Å)