ArticleProtein engineering, design & selection : PEDS2014
Computationally designed libraries for rapid enzyme stabilization.
Article in Protein engineering, design & selection : PEDS, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 82 papers.
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Who cites it
82 citing papers in PubMed, 263 citations in OpenAlex.
- Encapsulation in a Bacterial Microcompartment Shell Improves Thermal Stability of a Glycolytic Enzyme.ACS synthetic biology · 2026Article
- Introduction to the Field of Enzyme Immobilization and Stabilization.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Characterization of Xyloglucanase TpXEG12a fromInternational journal of molecular sciences · 2025Article
- Enriching stabilizing mutations through automated analysis of molecular dynamics simulations using BoostMut.Protein science : a publication of the Protein Society · 2025Article
- Buttressing ligand-binding pockets in de novo designed NTF2-like domains.Protein science : a publication of the Protein Society · 2025Article
- Semirational Protein Engineering Yields Archaerhodopsin-3-Based Fluorescent Genetically Encoded Voltage Indicators with Enhanced Brightness and Red Shifted Absorption Bands.Chem & bio engineering · 2025Article
- Enhancing recombinant growth factor and serum protein production for cultivated meat manufacturing.Microbial cell factories · 2025Review
- A Practical Guide to Computational Tools for Engineering Biocatalytic Properties.International journal of molecular sciences · 2025Review
- Structure-based self-supervised learning enables ultrafast protein stability prediction upon mutation.Innovation (Cambridge (Mass.)) · 2025Article
- Enhancement of the Thermostability ofMolecules (Basel, Switzerland) · 2024Article
- GRAPE-WEB: An automated computational redesign web server for improving protein thermostability.mLife · 2024Article
- Solvent concentration at 50% protein unfolding may reform enzyme stability ranking and process window identification.Nature communications · 2024Article
- Biosynthetic production of anticoagulant heparin polysaccharides through metabolic and sulfotransferases engineering strategies.Nature communications · 2024Article
- Application of an alchemical free energy method for the prediction of thermostable DuraPETase variants.Applied microbiology and biotechnology · 2024Article
- Computational Enzyme Redesign Enhances Tolerance to Denaturants for Peptide C-Terminal Amidation.JACS Au · 2024Article
- Computational design of an imine reductase: mechanism-guided stereoselectivity reversion and interface stabilization.Chemical science · 2024Article
- Fixing Flavins: Hijacking a Flavin Transferase for Equipping Flavoproteins with a Covalent Flavin Cofactor.Journal of the American Chemical Society · 2023Article
- A novel computationally engineered collagenase reduces the force required for tooth extraction in an ex-situ porcine jaw model.Journal of biological engineering · 2023Article
- A high-performance electrochemical biosensor using an engineered urate oxidase.Chemical communications (Cambridge, England) · 2023Article
- Recent Advances in β-Glucosidase Sequence and Structure Engineering: A Brief Review.Molecules (Basel, Switzerland) · 2023Review
22 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The ability to engineer enzymes and other proteins to any desired stability would have wide-ranging applications. Here, we demonstrate that computational design of a library with chemically diverse stabilizing mutations allows the engineering of drastically stabilized and fully functional variants of the mesostable enzyme limonene epoxide hydrolase. First, point mutations were selected if they significantly improved the predicted free energy of protein folding. Disulfide bonds were designed using sampling of backbone conformational space, which tripled the number of experimentally stabilizing disulfide bridges. Next, orthogonal in silico screening steps were used to remove chemically unreasonable mutations and mutations that are predicted to increase protein flexibility. The resulting library of 64 variants was experimentally screened, which revealed 21 (pairs of) stabilizing mutations located both in relatively rigid and in flexible areas of the enzyme. Finally, combining 10-12 of these confirmed mutations resulted in multi-site mutants with an increase in apparent melting temperature from 50 to 85°C, enhanced catalytic activity, preserved regioselectivity and a >250-fold longer half-life. The developed Framework for Rapid Enzyme Stabilization by Computational libraries (FRESCO) requires far less screening than conventional directed evolution.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.