ArticleNature communications2021
Versatile selective evolutionary pressure using synthetic defect in universal metabolism.
Article in Nature communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Crosstalk Between Lysine Lactylation and Acetylation Regulates Lactate Dehydrogenase in Streptococcus mutans.Genomics, proteomics & bioinformatics · 2025Article
- Directed evolution of hydrocarbon-producing enzymes.Biotechnology for biofuels and bioproducts · 2025Review
- High-throughput metabolic engineering ofProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Construction and Characterization of MoClo-Compatible Vectors for Modular Protein Expression inACS synthetic biology · 2025Article
- An aldolase-dependent phloroglucinol degradation pathway inApplied and environmental microbiology · 2024Article
- Growth-Coupled Evolutionary Pressure Improving Epimerases for D-Allulose Biosynthesis Using a Biosensor-Assisted In Vivo Selection Platform.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Advances in ligand-specific biosensing for structurally similar molecules.Cell systems · 2023Review
- A primer to directed evolution: current methodologies and future directions.RSC chemical biology · 2023Review
- Biotechnological production of omega-3 fatty acids: current status and future perspectives.Frontiers in microbiology · 2023Review
- A growth selection system for the directed evolution of amine-forming or converting enzymes.Nature communications · 2022Article
- Orthogonal glycolytic pathway enables directed evolution of noncanonical cofactor oxidase.Nature communications · 2022Article
- YiaC and CobB regulate lysine lactylation in Escherichia coli.Nature communications · 2022Article
- Directed evolution of phosphite dehydrogenase to cycle noncanonical redox cofactors via universal growth selection platform.Nature communications · 2022Article
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Authors and funding
3 authors.
Funding
Abstract
The non-natural needs of industrial applications often require new or improved enzymes. The structures and properties of enzymes are difficult to predict or design de novo. Instead, semi-rational approaches mimicking evolution entail diversification of parent enzymes followed by evaluation of isolated variants. Artificial selection pressures coupling desired enzyme properties to cell growth could overcome this key bottleneck, but are usually narrow in scope. Here we show diverse enzymes using the ubiquitous cofactors nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP) can substitute for defective NAD regeneration, representing a very broadly-applicable artificial selection. Inactivation of Escherichia coli genes required for anaerobic NAD regeneration causes a conditional growth defect. Cells are rescued by foreign enzymes connected to the metabolic network only via NAD or NADP, but only when their substrates are supplied. Using this principle, alcohol dehydrogenase, imine reductase and nitroreductase variants with desired selectivity modifications, and a high-performing isopropanol metabolic pathway, are isolated from libraries of millions of variants in single-round experiments with typical limited information to guide design.
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