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

Qu, G., Li, A., Acevedo-Rocha, C. G., Sun, Z., & Reetz, M. T. (2020). The Crucial Role of Methodology Development in Directed Evolution of Selective Enzymes. Angewandte Chemie, International Edition, 59(32), 13204-13231. doi:10.1002/anie.201901491.

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 Creators:
Qu, Ge1, Author
Li, Aitao2, Author
Acevedo-Rocha, Carlos G.3, Author
Sun, Zhoutong1, Author
Reetz, Manfred T.1, 4, 5, Author           
Affiliations:
1Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308 China, ou_persistent22              
2State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei University, 368 Youyi Road, Wuchang Wuhan, 430062 China, ou_persistent22              
3Biosyntia ApS, 2100 Copenhagen, Denmark, ou_persistent22              
4Research Department Reetz, Max-Planck-Institut für Kohlenforschung, Max Planck Society, ou_1445588              
5Department of Chemistry, Hans-Meerwein-Strasse 4, Philipps-University, 35032 Marburg, Germany, ou_persistent22              

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Free keywords: directed evolution; iterative saturation mutagenesis; machine learning; stereoselectivity; synthetic genes
 Abstract: Directed evolution of stereo‐, regio‐ and chemoselective enzymes provides a means to generate biocatalysts for synthetically interesting transformations. In order for this protein engineering technique to be efficient, fast and reliable, and of relevance to organic chemistry, methodology development was and still is necessary. Following a description of early contributions, this review focuses on recent developments originating from various groups. It includes optimization of molecular biological methods for gene mutagenesis and the design of efficient strategies for their application, resulting in notable reduction of the screening effort (bottleneck of directed evolution). When aiming for laboratory evolution of selectivity and activity, the newest versions of Combinatorial Active‐Site Saturation Test (CAST) and Iterative Saturation Mutagenesis (ISM) have emerged as preferred approaches, flanked by in silico methods such as machine learning. On‐chip solid‐phase chemical gene synthesis for rapid library construction enhances library quality notably, the future of directed evolution?

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Language(s): eng - English
 Dates: 2019-02-022019-07-022020-08-03
 Publication Status: Published in print
 Pages: 28
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/anie.201901491
 Degree: -

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Title: Angewandte Chemie, International Edition
  Abbreviation : Angew. Chem., Int. Ed.
Source Genre: Journal
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Publ. Info: Weinheim : Wiley-VCH
Pages: - Volume / Issue: 59 (32) Sequence Number: - Start / End Page: 13204 - 13231 Identifier: ISSN: 1433-7851
CoNE: https://pure.mpg.de/cone/journals/resource/1433-7851