ArticleACS synthetic biology2018
Development of a High-Throughput, In Vivo Selection Platform for NADPH-Dependent Reactions Based on Redox Balance Principles.
Article in ACS synthetic biology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 42 citations in OpenAlex.
- Effects of heterologous expression of glycolysis enzymes on product formation inMetabolic engineering communications · 2026Article
- Nitric Oxide Inhibition of Glycyl Radical Enzymes and Their Activases.Journal of the American Chemical Society · 2025Article
- A growth-based screening strategy for engineering the catalytic activity of an oxygen-sensitive formate dehydrogenase.Applied and environmental microbiology · 2024Article
- Strangers in a foreign land: 'Yeastizing' plant enzymes.Microbial biotechnology · 2024Review
- Automated in vivo enzyme engineering accelerates biocatalyst optimization.Nature communications · 2024Review
- Orthogonal glycolytic pathway enables directed evolution of noncanonical cofactor oxidase.Nature communications · 2022Article
- Directed evolution of phosphite dehydrogenase to cycle noncanonical redox cofactors via universal growth selection platform.Nature communications · 2022Article
- A phosphite-based screening platform for identification of enzymes favoring nonnatural cofactors.Scientific reports · 2022Article
- Increasing NADPH impairs fungal HBioresources and bioprocessing · 2022Article
- Can thiol-based redox systems be utilized as parts for synthetic biology applications?Redox report : communications in free radical research · 2021Article
- Growth-Based, High-Throughput Selection for NADH Preference in an Oxygen-Dependent Biocatalyst.ACS synthetic biology · 2021Article
- Growth-coupled selection of synthetic modules to accelerate cell factory development.Nature communications · 2021Article
- Article
- "NAD-display": Ultrahigh-Throughput in Vitro Screening of NAD(H) Dehydrogenases Using Bead Display and Flow Cytometry.Angewandte Chemie (International ed. in English) · 2021Article
- Engineering natural and noncanonical nicotinamide cofactor-dependent enzymes: design principles and technology development.Current opinion in biotechnology · 2020Review
- Leveraging Oxidative Stress to Regulate Redox Balance-Based,ACS synthetic biology · 2020Article
- In Vivo, High-Throughput Selection of Thermostable Cyclohexanone Monooxygenase (CHMO).Catalysts (Basel, Switzerland) · 2020Article
- A Growth-Based, High-Throughput Selection Platform Enables Remodeling of 4-Hydroxybenzoate Hydroxylase Active Site.ACS catalysis · 2020Article
- Redox Systems Biology: Harnessing the Sentinels of the Cysteine Redoxome.Antioxidants & redox signaling · 2020Review
- Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis.Nature chemical biology · 2020Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Bacteria undergoing anaerobic fermentation must maintain redox balance. In vivo metabolic evolution schemes based on this principle have been limited to targeting NADH-dependent reactions. Here, we developed a facile, specific, and high-throughput growth-based selection platform for NADPH-consuming reactions in vivo, based on an engineered NADPH-producing glycolytic pathway in Escherichia coli. We used the selection system in the directed evolution of a NADH-dependent d-lactate dehydrogenase from Lactobacillus delbrueckii toward utilization of NADPH. Through one round of selection, we obtained multiple enzyme variants with superior NADPH-dependent activities and protein expression levels; these mutants may serve as important tools in biomanufacturing d-lactate as a renewable polymer building block. Importantly, sequence analysis and computational protein modeling revealed that diverging evolutionary paths during the selection resulted in two distinct cofactor binding modes, which suggests that the high throughput of our selection system allowed deep searching of protein sequence space to discover diverse candidates en masse.
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