High-throughput colorimetric assays optimized for detection of ketones and aldehydes produced by microbial cell factories.

ABSTRACT: Randomized strain and pathway engineering are critical to improving microbial cell factory performance, calling for the development of high-throughput screening and selection systems. To facilitate this effort, we have developed two 96-well plate format colorimetric assays for reliable quantification of various ketones and aldehydes from culture supernatants, based on either a vanillin-acetone reaction or the 2,4-dinitrophenylhydrazine (2,4-DNPH) reagent. The vanillin-acetone assay enabled accurate and selective measurement of acetone titers up to 2 g l^-1 in a minimal culture medium. The 2,4-DNPH-based assay can be used for a wide range of aldehydes and ketones, shown here through the optimization of conditions for 15 different compounds. Both assays were implemented to improve

SUBMITTER: Kozaeva E 

PROVIDER: S-EPMC9437884 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature