Directed evolution of prenylated FMN-dependent Fdc supports efficient in vivo isobutene production

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 decarbox...

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Bibliographic Details
Published in:Nature communications 2021-09, Vol.12 (1), p.5300-5300, Article 5300
Main Authors: Saaret, Annica, Villiers, Benoît, Stricher, François, Anissimova, Macha, Cadillon, Mélodie, Spiess, Reynard, Hay, Sam, Leys, David
Format: Article
Language:English
Subjects:
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Acids
Acrylic acid
Computer applications
Conjugation
Crystal structure
Decarboxylation
Directed evolution
Enzymes
Evolution
Ferulic acid
Fossil fuels
Fuel additives
Humanities and Social Sciences
Mevalonate pathway
Mevalonic acid
multidisciplinary
Science
Science (multidisciplinary)
Selectivity
Substrates
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Summary: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. Isobutene is a high value gaseous alkene that is widely used as fuel additive and a chemical building block. Here, the authors report an alternative pathway for isobutene bioproduction by directed evolution of prenylated FMN-dependent ferulic acid decarboxylase.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-021-25598-0