Fúster Fernández, Inés ; Kickstein, Michael ; Fessner, Wolf‐Dieter (2023)
Hydroxamate assays for high‐throughput screening of transketolase libraries against arylated substrates.
In: Advanced Synthesis & Catalysis, 365 (22)
doi: 10.1002/adsc.202300989
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (Abstract)
We recently reported that the transketolase from Geobacillus stearothermophilus (TKgst) upon acyl transfer to nitrosoarenes generates N‐aryl hydroxamic acids (HA). The latter are metal chelating compounds that in the presence of Fe(III) ions form deep‐red complexes. Here, we applied this principle to the development of a colorimetric assay in both solid‐ and liquid‐phase formats for the high‐throughput screening of TKgst and its variants. Screening a set of positive hits from a L382X/D470X library validated the specificity and sensitivity of the assays. The solid surface assay allows a clear distinction between positive and negative colonies by the naked eye in qualitative mode, and further also to measure activity in semi‐quantitative fashion in the liquid‐phase format. The assay will be important for engineering the TKgst enzyme towards improved conversion of aromatic aldehydes by their close structural analogy to nitrosoarenes.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2023 |
| Autor(en): | Fúster Fernández, Inés ; Kickstein, Michael ; Fessner, Wolf‐Dieter |
| Art des Eintrags: | Bibliographie |
| Titel: | Hydroxamate assays for high‐throughput screening of transketolase libraries against arylated substrates |
| Sprache: | Englisch |
| Publikationsjahr: | 21 November 2023 |
| Ort: | Weinheim |
| Verlag: | Wiley-VCH |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Advanced Synthesis & Catalysis |
| Jahrgang/Volume einer Zeitschrift: | 365 |
| (Heft-)Nummer: | 22 |
| DOI: | 10.1002/adsc.202300989 |
| Zugehörige Links: | |
| Kurzbeschreibung (Abstract): | We recently reported that the transketolase from Geobacillus stearothermophilus (TKgst) upon acyl transfer to nitrosoarenes generates N‐aryl hydroxamic acids (HA). The latter are metal chelating compounds that in the presence of Fe(III) ions form deep‐red complexes. Here, we applied this principle to the development of a colorimetric assay in both solid‐ and liquid‐phase formats for the high‐throughput screening of TKgst and its variants. Screening a set of positive hits from a L382X/D470X library validated the specificity and sensitivity of the assays. The solid surface assay allows a clear distinction between positive and negative colonies by the naked eye in qualitative mode, and further also to measure activity in semi‐quantitative fashion in the liquid‐phase format. The assay will be important for engineering the TKgst enzyme towards improved conversion of aromatic aldehydes by their close structural analogy to nitrosoarenes. |
| Freie Schlagworte: | transketolase, hydroxamic acids, iron (III) chelation, high-throughput screening, colorimetric assay |
| Zusätzliche Informationen: | This article also appears in: Hot Topic: Automated Synthesis |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie |
| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Organische Chemie |
| Hinterlegungsdatum: | 28 Mai 2024 06:48 |
| Letzte Änderung: | 28 Mai 2024 12:36 |
| PPN: | 518692884 |
| Export: | |
| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
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Hydroxamate Assays for High‐Throughput Screening of Transketolase Libraries Against Arylated Substrates. (deposited 27 Mai 2024 13:08)
- Hydroxamate assays for high‐throughput screening of transketolase libraries against arylated substrates. (deposited 28 Mai 2024 06:48) [Gegenwärtig angezeigt]
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