ArticleScientific reports2017
Saturated mutagenesis of ketoisovalerate decarboxylase V461 enabled specific synthesis of 1-pentanol via the ketoacid elongation cycle.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 42 citations in OpenAlex.
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- Engineering New-to-Nature Biological Pathways for β,γ-Alkanediol Synthesis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Engineering of 2-ketoacid Decarboxylases for Production of Isobutanol and Other Fusel Alcohols in Saccharomyces cerevisiae.Biotechnology and bioengineering · 2026Article
- Directed evolution of α-ketoisovalerate decarboxylase for improved isobutanol and 3-methyl-1-butanol production in cyanobacteria.Biotechnology for biofuels and bioproducts · 2025Article
- Model-Guided Rational Construction of Escherichia coli Synthetic Consortia for Enhanced 2-Methylbutyric Acid Production.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Recent advances in microbial 3-methyl-1-butanol production.Frontiers in microbiology · 2025Review
- Production of (R)-citramalate by engineeredMetabolic engineering communications · 2024Article
- Novel Bioproduction of 1,6-Hexamethylenediamine from l-Lysine Based on an Artificial One-Carbon Elongation Cycle.ACS omega · 2024Article
- Decarboxylation in Natural Products Biosynthesis.JACS Au · 2024Review
- Silver Lactoferrin as Antimicrobials: Mechanisms of Action and Resistance Assessed by Bacterial Molecular Profiles.ACS omega · 2023Article
- Rational protein engineering of a ketoacids decarboxylase for efficient production of 1,2,4-butanetriol from arabinose.Biotechnology for biofuels and bioproducts · 2023Article
- Advances in biosynthesis of higher alcohols in Escherichia coli.World journal of microbiology & biotechnology · 2023Review
- Rebooting life: engineering non-natural nucleic acids, proteins and metabolites in microorganisms.Microbial cell factories · 2022Review
- A Critical Review on the Economically Feasible and Sustainable Poly(3-Hydroxybutyrate-Polymers · 2022Review
- A High-Efficiency Artificial Synthetic Pathway for 5-Aminovalerate Production From Biobased L-Lysine inFrontiers in bioengineering and biotechnology · 2021Article
- Engineering Improves Enzymatic Synthesis of L-Tryptophan by Tryptophan Synthase fromMicroorganisms · 2020Article
- Synthetic Biology and Metabolic Engineering EmployingFrontiers in bioengineering and biotechnology · 2020Review
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Iterative ketoacid elongation has been an essential tool in engineering artificial metabolism, in particular the synthetic alcohols. However, precise control of product specificity is still greatly challenged by the substrate promiscuity of the ketoacid decarboxylase, which unselectively hijacks ketoacid intermediates from the elongation cycle along with the target ketoacid. In this work, preferential tuning of the Lactococcus lactis ketoisovalerate decarboxylase (Kivd) specificity toward 1-pentanol synthesis was achieved via saturated mutagenesis of the key residue V461 followed by screening of the resulting alcohol spectrum. Substitution of V461 with the small and polar amino acid glycine or serine significantly improved the Kivd selectivity toward the 1-pentanol precursor 2-ketocaproate by lowering its catalytic efficiency for the upstream ketoacid 2-ketobutyrate and 2-ketovalerate. Conversely, replacing V461 with bulky or charged side chains displayed severely adverse effect. Increasing supply of the iterative addition unit acetyl-CoA by acetate feeding further drove 2-ketoacid flux into the elongation cycle and enhanced 1-pentanol productivity. The Kivd V461G variant enabled a 1-pentanol production specificity around 90% of the total alcohol content with or without oleyl alcohol extraction. This work adds insight to the selectivity of Kivd active site.
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