7NF0

T. atroviride Fdc variant TaFdcV in complex with hydroxylated prFMN

Summary for 7NF0

• Entry DOI: 10.2210/pdb7nf0/pdb
• Descriptor: Ferulic acid decarboxylase 1, NITRATE ION, hydroxylated prenyl-FMN, ... (7 entities in total)
• Functional Keywords: fdc, prfmn, decarboxylase, isobutene production, lyase
• Biological source: Hypocrea atroviridis (strain ATCC 20476 / IMI 206040)
• Total number of polymer chains: 2
• Total formula weight: 115511.89

Authors

Saaret, A.,Leys, D. (deposition date: 2021-02-05, release date: 2021-08-04, Last modification date: 2024-01-31)

Primary citation

Saaret, A.,Villiers, B.,Stricher, F.,Anissimova, M.,Cadillon, M.,Spiess, R.,Hay, S.,Leys, D.
Directed evolution of prenylated FMN-dependent Fdc supports efficient in vivo isobutene production.
Nat Commun, 12:5300-5300, 2021

PubMed Abstract: Isobutene is a high value gaseous alkene used as fuel additive and a chemical building block. As an alternative to fossil fuel derived isobutene, we here develop a modified mevalonate pathway for the production of isobutene from glucose in vivo. The final step in the pathway consists of the decarboxylation of 3-methylcrotonic acid, catalysed by an evolved ferulic acid decarboxylase (Fdc) enzyme. Fdc belongs to the prFMN-dependent UbiD enzyme family that catalyses reversible decarboxylation of (hetero)aromatic acids or acrylic acids with extended conjugation. Following a screen of an Fdc library for inherent 3-methylcrotonic acid decarboxylase activity, directed evolution yields variants with up to an 80-fold increase in activity. Crystal structures of the evolved variants reveal that changes in the substrate binding pocket are responsible for increased selectivity. Solution and computational studies suggest that isobutene cycloelimination is rate limiting and strictly dependent on presence of the 3-methyl group.

PubMed: 34489427
DOI: 10.1038/s41467-021-25598-0

Experimental method

X-RAY DIFFRACTION (1.35 Å)