Directed Evolution: Methodologies and Applications

Chemical Reviews, Journal Year: 2021, Volume and Issue: 121(20), P. 12384 - 12444

Published: July 23, 2021

Directed evolution aims to expedite the natural process of biological molecules and systems in a test tube through iterative rounds gene diversifications library screening/selection. It has become one most powerful widespread tools for engineering improved or novel functions proteins, metabolic pathways, even whole genomes. This review describes commonly used diversification strategies, screening/selection methods, recently developed continuous strategies directed evolution. Moreover, we highlight some representative applications nucleic acids, genetic circuits, viruses, cells. Finally, discuss challenges future perspectives

Language: Английский

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The Crucial Role of Methodology Development in Directed Evolution of Selective Enzymes

Angewandte Chemie International Edition, Journal Year: 2019, Volume and Issue: 59(32), P. 13204 - 13231

Published: July 3, 2019

Directed evolution of stereo-, regio-, and chemoselective enzymes constitutes a unique way to generate biocatalysts for synthetically interesting transformations in organic chemistry biotechnology. In order this protein engineering technique be efficient, fast, reliable, also relevance synthetic chemistry, methodology development was still is necessary. Following description early key contributions, review focuses on recent developments. It includes optimization molecular biological methods gene mutagenesis the design efficient strategies their application, resulting notable reduction screening effort (bottleneck directed evolution). When aiming laboratory selectivity activity, second-generation versions Combinatorial Active-Site Saturation Test (CAST) Iterative Mutagenesis (ISM), both involving saturation (SM) at sites lining binding pocket, have emerged as preferred approaches, aided by silico such machine learning. The recently proposed Focused Rational Site-specific (FRISM) fusion rational evolution. On-chip solid-phase chemical synthesis rapid library construction enhances quality notably eliminating undesired amino acid bias, future evolution?

Language: Английский

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Enantioselective [2+2]-cycloadditions with triplet photoenzymes

Nature, Journal Year: 2022, Volume and Issue: 611(7937), P. 715 - 720

Published: Sept. 21, 2022

Language: Английский

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Photoinduced chemomimetic biocatalysis for enantioselective intermolecular radical conjugate addition

Nature Catalysis, Journal Year: 2022, Volume and Issue: 5(7), P. 586 - 593

Published: May 2, 2022

Language: Английский

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Thermal stability enhancement: Fundamental concepts of protein engineering strategies to manipulate the flexible structure

International Journal of Biological Macromolecules, Journal Year: 2022, Volume and Issue: 214, P. 642 - 654

Published: June 27, 2022

Language: Английский

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A light-driven enzymatic enantioselective radical acylation

Nature, Journal Year: 2023, Volume and Issue: 625(7993), P. 74 - 78

Published: Dec. 18, 2023

Language: Английский

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Navigating the landscape of enzyme design: from molecular simulations to machine learning

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(16), P. 8202 - 8239

Published: Jan. 1, 2024

Global environmental issues and sustainable development call for new technologies fine chemical synthesis waste valorization. Biocatalysis has attracted great attention as the alternative to traditional organic synthesis. However, it is challenging navigate vast sequence space identify those proteins with admirable biocatalytic functions. The recent of deep-learning based structure prediction methods such AlphaFold2 reinforced by different computational simulations or multiscale calculations largely expanded 3D databases enabled structure-based design. While approaches shed light on site-specific enzyme engineering, they are not suitable large-scale screening potential biocatalysts. Effective utilization big data using machine learning techniques opens up a era accelerated predictions. Here, we review applications machine-learning guided We also provide our view challenges perspectives effectively employing design integrating molecular learning, importance database construction algorithm in attaining predictive ML models explore fitness landscape

Language: Английский

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Biological modification and industrial applications of microbial lipases: A general review

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: unknown, P. 140486 - 140486

Published: Jan. 1, 2025

Language: Английский

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Design of novel enzyme biocatalysts for industrial bioprocess: Harnessing the power of protein engineering, high throughput screening and synthetic biology

Bioresource Technology, Journal Year: 2020, Volume and Issue: 325, P. 124617 - 124617

Published: Dec. 26, 2020

Language: Английский

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Stereodivergent Carbon–Carbon Bond Formation between Iminium and Enolate Intermediates by Synergistic Organocatalysis

Journal of the American Chemical Society, Journal Year: 2020, Volume and Issue: 143(1), P. 73 - 79

Published: Dec. 24, 2020

We report here a stereodivergent method for the Michael addition of aryl acetic acid esters to α,β-unsaturated aldehydes catalyzed by combination chiral pyrrolidine and Lewis base. This reaction proceeds through synergistic catalytic cycle which consists one leading iminium electrophile second generating nucleophilic enolate construction carbon–carbon bond. By varying combinations catalyst enantiomers, all four stereoisomers products with two vicinal stereocenters are accessible high enantio- diastereoselectivity. The addition, 1,5-aldehyde esters, can be readily transformed into variety other valuable enantioenriched structures, including those bearing three contiguous in an acyclic system, thus providing efficient route array structural stereochemical diversity.

Language: Английский

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