6H6V

Structure of the UbiD-class enzyme HmfF from Pelotomaculum thermopropionicum in complex with FMN

Summary for 6H6V

• Entry DOI: 10.2210/pdb6h6v/pdb
• Descriptor: 3-polyprenyl-4-hydroxybenzoate decarboxylase and related decarboxylases, FLAVIN MONONUCLEOTIDE, POTASSIUM ION, ... (6 entities in total)
• Functional Keywords: prfmn, decarboxylation, thermophilic, ubid, hmff, flavoprotein
• Biological source: Pelotomaculum thermopropionicum SI
• Total number of polymer chains: 6
• Total formula weight: 297793.01

Authors

Payne, K.A.P.,Leys, D. (deposition date: 2018-07-30, release date: 2019-02-27, Last modification date: 2024-10-23)

Primary citation

Payne, K.A.P.,Marshall, S.A.,Fisher, K.,Cliff, M.J.,Cannas, D.M.,Yan, C.,Heyes, D.J.,Parker, D.A.,Larrosa, I.,Leys, D.
Enzymatic Carboxylation of 2-Furoic Acid Yields 2,5-Furandicarboxylic Acid (FDCA).
Acs Catalysis, 9:2854-2865, 2019

PubMed Abstract: The biological production of FDCA is of considerable value as a potential replacement for petrochemical-derived monomers such as terephthalate, used in polyethylene terephthalate (PET) plastics. HmfF belongs to an uncharacterized branch of the prenylated flavin (prFMN) dependent UbiD family of reversible (de)carboxylases and is proposed to convert 2,5-furandicarboxylic acid (FDCA) to furoic acid in vivo. We present a detailed characterization of HmfF and demonstrate that HmfF can catalyze furoic acid carboxylation at elevated CO levels in vitro. We report the crystal structure of a thermophilic HmfF from , revealing that the active site located above the prFMN cofactor contains a furoic acid/FDCA binding site composed of residues H296-R304-R331 specific to the HmfF branch of UbiD enzymes. Variants of the latter are compromised in activity, while H296N alters the substrate preference to pyrrole compounds. Solution studies and crystal structure determination of an engineered dimeric form of the enzyme revealed an unexpected key role for a UbiD family wide conserved Leu residue in activity. The structural insights into substrate and cofactor binding provide a template for further exploitation of HmfF in the production of FDCA plastic precursors and improve our understanding of catalysis by members of the UbiD enzyme family.

PubMed: 31057985
DOI: 10.1021/acscatal.8b04862

Experimental method

X-RAY DIFFRACTION (2.66 Å)