Complete Switch of Reaction Specificity of an Aldolase by Directed Evolution In Vitro: Synthesis of Generic Aliphatic Aldol Products
Abstract: A structure‐guided engineering of fructose‐6‐phosphate aldolase was performed to expand its substrate promiscuity toward aliphatic nucleophiles, that is, unsubstituted alkanones and alkanals. A “smart” combinatorial library was created targeting residues D6, T26, and N28, which form a binding pocket around the nucleophilic carbon atom. Double‐selectivity screening was executed by high‐performance TLC that allowed simultaneous determination of total activity as well as a preference for acetone versus propanal as competing nucleophiles. D6 turned out to be the key residue that enabled activity with non‐hydroxylated nucleophiles. Altogether 25 single‐ and double‐site variants (D6X and D6X/T26X) were discovered that show useful synthetic activity and a varying preference for ketone or aldehyde as the aldol nucleophiles. Remarkably, all of the novel variants had completely lost their native activity for cleavage of fructose 6‐phosphate.
- Standort
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Deutsche Nationalbibliothek Frankfurt am Main
- Umfang
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Online-Ressource
- Sprache
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Englisch
- Erschienen in
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Complete Switch of Reaction Specificity of an Aldolase by Directed Evolution In Vitro: Synthesis of Generic Aliphatic Aldol Products ; volume:57 ; number:32 ; year:2018 ; pages:10153-10157 ; extent:5
Angewandte Chemie / International edition. International edition ; 57, Heft 32 (2018), 10153-10157 (gesamt 5)
- Urheber
- DOI
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10.1002/anie.201804831
- URN
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urn:nbn:de:101:1-2022081608175379620869
- Rechteinformation
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Letzte Aktualisierung
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15.08.2025, 07:34 MESZ
Datenpartner
Deutsche Nationalbibliothek. Bei Fragen zum Objekt wenden Sie sich bitte an den Datenpartner.
Beteiligte
- Junker, Sebastian
- Roldan, Raquel
- Joosten, Henk‐Jan
- Clapés, Pere
- Fessner, Wolf-Dieter
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Direct Crossed-Aldol Reaction of Acetaldehyde
Homo-aldol Reactions of Unsaturated Carbamates
Biocatalytic Transamination of Aldolase‐Derived 3‐Hydroxy Ketones
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Thermal Properties of Aliphatic Polypeptoids
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