Protein Engineering by Structure‐Guided SCHEMA Recombination

Saab‐Rincón, Gloria; Li, Yougen; Meyer, Michelle M.; Carbone, Martina; Landwehr, Marco; Arnold, Frances H.

doi:10.1002/9783527634026.ch20

Cited by 3 publications

(3 citation statements)

References 32 publications

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“…During the last five years this picture has changed, because researchers began to consider "quality not quantity", [59g] which signaled the call for methodology development in directed evolution. [4,18,31,[58][59][60][61][62][63] It is not just industry that is in need of "fast" and practical protein engineering methods, basic research leads the way. [4] Our contribution regarding this crucial goal is iterative saturation mutagenesis (ISM) as a means to control the stereoselectivity [18,31,[58][59][60] and the substrate scope of enzymes, [18,58] as well as to increase the thermostability [20,77,120] of proteins in general.…”

Section: Discussionmentioning

confidence: 99%

“…[1] We have defined the quality of a mutant library in terms of the frequency of hits and their degree of catalyst improvement, [58,60] which in turn touches on the screening problem. [10] Some research groups have utilized various computational aids in this endeavor, with prominent examples being SCHEMA, [61] ProSAR, [62] FamClash, [63a] GLUE-IT and PEDEL-AA, [63b] and others [63c-h] such as CASTER and B-FITTER developed in my research group. [20,60] In this Review, the focus is primarily on the development of particularly effective methods in directed evolution for controlling stereoselectivity, substrate scope, and thermostability.…”

Section: Early Examples Of Directed Evolution Of Enantioselective Enzymesmentioning

confidence: 99%

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Laboratory Evolution of Stereoselective Enzymes: A Prolific Source of Catalysts for Asymmetric Reactions

2010

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Asymmetric catalysis plays a key role in modern synthetic organic chemistry, with synthetic catalysts and enzymes being the two available options. During the latter part of the last century the use of enzymes in organic chemistry and biotechnology experienced a period of rapid growth. However, these biocatalysts have traditionally suffered from several limitations, including in many cases limited substrate scope, poor enantioselectivity, insufficient stability, and sometimes product inhibition. During the last 15 years, the genetic technique of directed evolution has been developed to such an extent that all of these long-standing problems can be addressed and solved. It is based on repeated cycles of gene mutagenesis, expression, and screening (or selection). This Review focuses on the directed evolution of enantioselective enzymes, which constitutes a fundamentally new approach to asymmetric catalysis. Emphasis is placed on the development of methods to make laboratory evolution faster and more efficient, thus providing chemists and biotechnologists with a rich and non-ending source of robust and selective catalysts for a variety of useful applications.

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Section: Discussionmentioning

confidence: 99%

Section: Early Examples Of Directed Evolution Of Enantioselective Enzymesmentioning

confidence: 99%

Laboratory Evolution of Stereoselective Enzymes: A Prolific Source of Catalysts for Asymmetric Reactions

2010

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“…Vergleich der Kristallstruktur von PAMO [39] (links) mit dem Homologiemodell von CHMO [40] [61] ProSAR, [62] FamClash, [63a] GLUE-IT und PEDEL-AA [63b] und andere [63c-h] sowie die von uns vorgestellten Verfahren CASTER und B-FITTER. [20,60] In diesem Aufsatz liegt der Schwerpunkt auf unseren Beiträgen zur Entwicklung effizienterer Methoden in der gerichteten Evolution, insbesondere zur Kontrolle der Stereoselektivität und der Substratbreite (Aktivität).…”

Section: Erste Beispiele Für Gerichtete Evolution Enantioselektiver Eunclassified

Gerichtete Evolution stereoselektiver Enzyme: Eine ergiebige Katalysator‐Quelle für asymmetrische Reaktionen

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2010

Angewandte Chemie

130

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Asymmetrische Katalyse spielt eine Schlüsselrolle in der modernen synthetischen organischen Chemie, wobei künstliche Katalysatoren und Enzyme die zwei wichtigsten Optionen darstellen. Die Verwendung von Enzymen in der organischen Chemie und Biotechnologie hat in der zweiten Hälfte des vergangenen Jahrhunderts stark zugenommen, oft blieb jedoch der Anwendungsbereich aus mehreren Gründen eingeschränkt. Dazu gehören begrenzte Substratakzeptanz, geringe oder fehlende Stereoselektivität, unzureichende Stabilität und manchmal Produktinhibierung. Während der vergangenen 15 Jahre wurde die genetische Technik der gerichteten Evolution so weit entwickelt und verfeinert, dass all diese Probleme angegangen und in der Regel gelöst werden können. Sie basiert auf wiederholende Zyklen von Genmutagenese, Expression und Screening (oder Selektion). Der vorliegende Aufsatz fokussiert auf gerichtete Evolution stereoselektiver Enzyme, ein fundamental neuer Ansatz zur asymmetrischen Katalyse. Der Schwerpunkt liegt auf Methodenentwicklung in dem Bestreben, diese Art der Katalysator‐Optimierung einfacher, schneller und effizienter als in der Vergangenheit zu gestalten. Die neueren Methoden bescheren dem Chemiker und dem Biotechnologen robuste und selektive Katalysatoren für eine Vielzahl von nützlichen Anwendungen.

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Using the Evolutionary History of Proteins to Engineer Insertion-Deletion Mutants from Robust, Ancestral Templates Using Graphical Representation of Ancestral Sequence Predictions (GRASP)

Ross

Foley

Bodén

et al. 2021

Methods in Molecular Biology

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Protein Engineering by Structure‐Guided SCHEMA Recombination

Cited by 3 publications

References 32 publications

Laboratory Evolution of Stereoselective Enzymes: A Prolific Source of Catalysts for Asymmetric Reactions

Laboratory Evolution of Stereoselective Enzymes: A Prolific Source of Catalysts for Asymmetric Reactions

Gerichtete Evolution stereoselektiver Enzyme: Eine ergiebige Katalysator‐Quelle für asymmetrische Reaktionen

Using the Evolutionary History of Proteins to Engineer Insertion-Deletion Mutants from Robust, Ancestral Templates Using Graphical Representation of Ancestral Sequence Predictions (GRASP)

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