Substrate-Independent High-Throughput Assay for the Quantification of Aldehydes

Anna K. Ressmann, Daniel Schwendenwein, Simon Leonhartsberger, Marko D. Mihovilovic, Uwe T. Bornscheuer, Margit Winkler, Florian Rudroff

Research output: Contribution to journalArticle

Abstract

The selective and direct reduction of carboxylic acids into the corresponding aldehydes by chemical methods is still a challenging task in synthesis. Several reductive and oxidative chemical methods are known to produce aldehydes, but most of them require expensive reagents, special reaction conditions, are two-step procedures and often lack chemoselectivity. Nature provides an elegant tool, so called carboxylic acid reductases (CARs) for the direct reduction of carboxylic acids to aldehydes. Discovery as well as engineering of novel CAR enzymes necessitates a robust, product selective high-throughput assay (HTA). We report a simple and fast HTA that allows the substrate-independent and chemoselective quantification of aldehydes (irrespective of their chemical structure) and is sensitive to the nM range. The HTA was validated by NMR and GC analyses and in microbial cells by reexamination of the substrate scope of CAR from Nocardia iowensis (CAR Ni ). The results were fully consistent with reported data. (Figure presented.).

Original languageEnglish
Pages (from-to)2538-2543
Number of pages6
JournalAdvanced Synthesis & Catalysis
Volume361
Issue number11
Early online date2019
DOIs
Publication statusPublished - 6 Jul 2019

Keywords

  • dihydroquinazoline
  • high-throughput screening
  • 2-amino-benzamidoxime
  • aldehyde quantification
  • cell compatible assay

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

Fields of Expertise

  • Human- & Biotechnology

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