TY - JOUR
T1 - Substrate-Independent High-Throughput Assay for the Quantification of Aldehydes
AU - Ressmann, Anna K.
AU - Schwendenwein, Daniel
AU - Leonhartsberger, Simon
AU - Mihovilovic, Marko D.
AU - Bornscheuer, Uwe T.
AU - Winkler, Margit
AU - Rudroff, Florian
PY - 2019/7/6
Y1 - 2019/7/6
N2 - 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.).
AB - 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.).
KW - dihydroquinazoline
KW - high-throughput screening
KW - 2-amino-benzamidoxime
KW - aldehyde quantification
KW - cell compatible assay
UR - http://www.scopus.com/inward/record.url?scp=85064075596&partnerID=8YFLogxK
U2 - 10.1002/adsc.201900154
DO - 10.1002/adsc.201900154
M3 - Article
SN - 1615-4150
VL - 361
SP - 2538
EP - 2543
JO - Advanced Synthesis & Catalysis
JF - Advanced Synthesis & Catalysis
IS - 11
ER -