Strategy for identification of -dihydrodiendiol-degrading dehydrogenases in BW25113.

-Dihydrodiendiols are valuable compounds, finding multiple application as chiral synthons in organic chemistry. The biotechnological route for the generation of -dihydrodiendiols involves the dihydroxylation of aromatic compounds, catalyzed by Rieske non-heme iron dioxygenases. To date, numerous examples of recombinant , harboring such dioxygenases, can be found in the literature. Nevertheless, there is only a minor number of publications, addressing the catalyzed degradation of -dihydrodiendiols into catechols dehydrogenases. Identification and elimination of such dehydrogenase catalyzed degradation is key for the establishment of enhanced recombinant platforms pursuing the production of -dihydrodiendiols. Here, we provide a fast and easy strategy for the identification of promiscuous alcohol dehydrogenases in BW25113, catalyzing the degradation of -dihydrodiendiols into catechols. This approach is based on the screening of dehydrogenase deficient KEIO strains, regarding their incapability of degrading a -dihydrodiendiol of choice.•Novel screening strategy for BW25113 dehydrogenase knock-outs, incapable of degrading -dihydrodiendiols was validated for -1,2-dihydrocatechol as substrate•Corresponding knock-outs can be used for recombinant production of -dihydrodiendiols•Simple analysis based on liquid chromatography with diode array detector (HPLC-DAD).