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Substrate‐Independent High‐Throughput Assay for the Quantification of Aldehydes
Author(s) -
Ressmann Anna K.,
Schwendenwein Daniel,
Leonhartsberger Simon,
Mihovilovic Marko D.,
Bornscheuer Uwe T.,
Winkler Margit,
Rudroff Florian
Publication year - 2019
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.201900154
Subject(s) - chemoselectivity , chemistry , substrate (aquarium) , reagent , carboxylic acid , combinatorial chemistry , aldehyde , high throughput screening , organic chemistry , catalysis , biochemistry , oceanography , geology
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.