Directed evolution of prenylated FMN-dependent Fdc supports efficient in vivo isobutene production.
Alkenes
/ chemistry
Biocatalysis
Carboxy-Lyases
/ genetics
Crotonates
/ metabolism
Directed Molecular Evolution
/ methods
Escherichia coli
/ enzymology
Escherichia coli Proteins
/ genetics
Fermentation
Flavin Mononucleotide
/ chemistry
Glucose
/ chemistry
Hypocreales
/ enzymology
Mevalonic Acid
/ metabolism
Prenylation
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
06 09 2021
06 09 2021
Historique:
received:
02
03
2021
accepted:
29
07
2021
entrez:
7
9
2021
pubmed:
8
9
2021
medline:
6
10
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 34489427
doi: 10.1038/s41467-021-25598-0
pii: 10.1038/s41467-021-25598-0
pmc: PMC8421414
doi:
Substances chimiques
Alkenes
0
Crotonates
0
Escherichia coli Proteins
0
Flavin Mononucleotide
7N464URE7E
Carboxy-Lyases
EC 4.1.1.-
phenylacrylic acid decarboxylase
EC 4.1.1.-
ubiX protein, E coli
EC 4.1.1.-
Glucose
IY9XDZ35W2
isobutylene
QA2LMR467H
Mevalonic Acid
S5UOB36OCZ
crotonic acid
YW5WZZ4O5Q
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5300Informations de copyright
© 2021. The Author(s).
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