Flow cytometry-based method for isolating live bacteria with meta-cleavage activity on dihydroxy compounds of biphenyl

摘要: A new method for isolating targeted live bacterial cells was established with the use of cell sorting by flow cytometry (FCM) based on fluorescence intermediate metabolite biphenyl degradation. During degradation, a PCB degrader, Comamonas testosteroni TK102, produces meta-cleavage metabolite, 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA), which emits green fluorescence. HOPDA produced from 2,3-dihydroxy as substrate, but it not appropriate labeling because released into medium. When we used 4-n-butylbiphenyl and 4-n-heptylbiphenyl, found that accumulated metabolites. By addition synthesized 2,3-dihydroxy-4′-butylbiphenyl (2,3-DHBBP), were able to label strong fluorescence, suggesting persistence fluorescent in introduction alkyl tail. 2,3-DHBBP then strain TK102 sorted FCM. The efficiency FCM defined percentage colony numbers per events. Strain successfully enriched 4.1-fold mixture environmental indigenous bacteria 7.3%. present here serves basic technique specific direct isolation contain dioxygenases active dihydroxylated aromatic compounds.