ArticleProceedings of the National Academy of Sciences of the United States of America2012
Combinatorial reshaping of the Candida antarctica lipase A substrate pocket for enantioselectivity using an extremely condensed library.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed.
- Deciphering the evolutionary origin of the enantioselectivity of short-chain dehydrogenases from plants toward 1-borneol.Nature communications · 2026Article
- Computational and experimental engineering of aFood chemistry: X · 2026Article
- Mechanistic Elucidation and Stereochemical Consequences of Alternative Binding of Alkenyl Substrates by Engineered Arylmalonate Decarboxylase.Journal of the American Chemical Society · 2025Article
- Easy and Versatile Technique for the Preparation of Stable and Active Lipase-Based CLEA-like Copolymers by Using Two Homofunctional Cross-Linking Agents: Application to the Preparation of Enantiopure Ibuprofen.International journal of molecular sciences · 2023Article
- Geometric Remodeling of Nitrilase Active Pocket Based on ALF-Scanning Strategy To Enhance Aromatic Nitrile Substrate Preference and Catalytic Efficiency.Applied and environmental microbiology · 2023Article
- Tuning Selectivity in CalA Lipase: Beyond Tunnel Engineering.Biochemistry · 2023Article
- Engineering of Cyclodextrin Glycosyltransferase through a Size/Polarity Guided Triple-Code Strategy with Enhanced α-Glycosyl Hesperidin Synthesis Ability.Applied and environmental microbiology · 2022Article
- Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis.Nature communications · 2022Article
- Lipase-catalyzed esterification in water enabled by nanomicelles. Applications to 1-pot multi-step sequences.Chemical science · 2022Article
- Learning Strategies in Protein Directed Evolution.Methods in molecular biology (Clifton, N.J.) · 2022Review
- Thermostable lipases and their dynamics of improved enzymatic properties.Applied microbiology and biotechnology · 2021Review
- Ethyl Butyrate Synthesis Catalyzed by Lipases A and B fromInternational journal of molecular sciences · 2019Article
- Direct enzymatic ethanolysis of potentialBiotechnology for biofuels · 2019Article
- Tunnel engineering to accelerate product release for better biomass-degrading abilities in lignocellulolytic enzymes.Biotechnology for biofuels · 2019Article
- Holistic engineering of Cal-A lipase chain-length selectivity identifies triglyceride binding hot-spot.PloS one · 2019Article
- Efficient molecular evolution to generate enantioselective enzymes using a dual-channel microfluidic droplet screening platform.Nature communications · 2018Article
- Specificity Effects of Amino Acid Substitutions in Promiscuous Hydrolases: Context-Dependence of Catalytic Residue Contributions to Local Fitness Landscapes in Nearby Sequence Space.Chembiochem : a European journal of chemical biology · 2017Article
- Integrated (Meta) Genomic and Synthetic Biology Approaches to Develop New Biocatalysts.Marine drugs · 2016Review
- Yeast Kluyveromyces lactis as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from Serratia sp.Journal of industrial microbiology & biotechnology · 2015Article
- Recombinant Lipases and Phospholipases and Their Use as Biocatalysts for Industrial Applications.International journal of molecular sciences · 2015Review
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5 authors.
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Abstract
A highly combinatorial structure-based protein engineering method for obtaining enantioselectivity is reported that results in a thorough modification of the substrate binding pocket of Candida antarctica lipase A (CALA). Nine amino acid residues surrounding the entire pocket were simultaneously mutated, contributing to a reshaping of the substrate pocket to give increased enantioselectivity and activity for a sterically demanding substrate. This approach seems to be powerful for developing enantioselectivity when a complete reshaping of the active site is required. Screening toward ibuprofen ester 1, a substrate for which previously used methods had failed, gave variants with a significantly increased enantioselectivity and activity. Wild-type CALA has a moderate activity with an E value of only 3.4 toward this substrate. The best variant had an E value of 100 and it also displayed a high activity. The variation at each mutated position was highly reduced, comprising only the wild type and an alternative residue, preferably a smaller one with similar properties. These minimal binary variations allow for an extremely condensed protein library. With this highly combinatorial method synergistic effects are accounted for and the protein fitness landscape is explored efficiently.
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