6F97

Crystal structure of the V465T mutant of 5-(Hydroxymethyl)furfural Oxidase (HMFO)

Summary for 6F97

• Entry DOI: 10.2210/pdb6f97/pdb
• Descriptor: 5-(hydroxymethyl)furfural oxidase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• Functional Keywords: sec-thiol oxidation, flavoprotein, alcohol oxidase, kinetic resolution, biocatalysis, enzyme engineering
• Biological source: Methylovorus sp. (strain MP688)
• Total number of polymer chains: 2
• Total formula weight: 115727.27

Authors

Pickl, M.,Swoboda, A.,Romero, E.,Winkler, C.K.,Binda, C.,Mattevi, A.,Faber, K.,Fraaije, M.W. (deposition date: 2017-12-14, release date: 2018-02-14, Last modification date: 2024-01-17)

Primary citation

Pickl, M.,Swoboda, A.,Romero, E.,Winkler, C.K.,Binda, C.,Mattevi, A.,Faber, K.,Fraaije, M.W.
Kinetic Resolution of sec-Thiols by Enantioselective Oxidation with Rationally Engineered 5-(Hydroxymethyl)furfural Oxidase.
Angew. Chem. Int. Ed. Engl., 57:2864-2868, 2018

PubMed Abstract: Various flavoprotein oxidases were recently shown to oxidize primary thiols. Herein, this reactivity is extended to sec-thiols by using structure-guided engineering of 5-(hydroxymethyl)furfural oxidase (HMFO). The variants obtained were employed for the oxidative kinetic resolution of racemic sec-thiols, thus yielding the corresponding thioketones and nonreacted R-configured thiols with excellent enantioselectivities (E≥200). The engineering strategy applied went beyond the classic approach of replacing bulky amino acid residues with smaller ones, as the active site was additionally enlarged by a newly introduced Thr residue. This residue established a hydrogen-bonding interaction with the substrates, as verified in the crystal structure of the variant. These strategies unlocked HMFO variants for the enantioselective oxidation of a range of sec-thiols.

PubMed: 29384246
DOI: 10.1002/anie.201713189

Experimental method

X-RAY DIFFRACTION (1.9 Å)