Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1002/cyto.a.20811
Title (Primary) NBDT (3-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-3-toluene) - A novel fluorescent dye for studying mechanisms of toluene uptake into vital bacteria
Author Sträuber, H.; Hübschmann, T.; Jehmlich, N. ORCID logo ; Schmidt, F.; von Bergen, M.; Harms, H.; Müller, S.
Source Titel Cytometry Part A
Year 2010
Volume 77A
Issue 2
Page From 113
Page To 120
Language englisch
Keywords Hydrophobic substrate uptake; NBD; dye efflux; XylN protein; vital bacteria; bacterial proteom
Abstract Uptake of small hydrophobic substances such as toluene into bacteria is widely assumed to occur by passive diffusion. Some toluene degrading bacteria, however, are described to contain uptake systems which may be involved in the transport of this compound. In this study, a fluorescently labeled toluene analogue dye (3-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-3-toluene; NBDT), flow cytometry, and shot gun proteome analysis were used to follow toluene uptake into bacteria in more detail. The new dye has excitation peaks at 444 and 475 nm and an emission peak at 537 nm. The toluene-degraders P. putida mt-2 and P. putida F1 as well as P. putida KT2440 and E. coli K12 as negative controls were included. To enable quantification of NBDT uptake, carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was added to inactivate NBDT efflux pumps. The porin inhibitor cadaverine was added to study the porin-mediated influx of toluene. Cadaverine reduced NBDT uptake by toluene-grown P. putida mt-2 and F1 by 25% and 42%, respectively, thus revealing an involvement and possibly a regulatory function of porins in the uptake of the toxic substrate toluene. Shot gun proteome measurements gave evidence for the presence of toluene transporting porins in P. putida mt-2 grown on toluene but not when grown on glucose. © 2009 International Society for Advancement of Cytometry
Persistent UFZ Identifier
Sträuber, H., Hübschmann, T., Jehmlich, N., Schmidt, F., von Bergen, M., Harms, H., Müller, S. (2010):
NBDT (3-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-3-toluene) - A novel fluorescent dye for studying mechanisms of toluene uptake into vital bacteria
Cytom. Part A 77A (2), 113 - 120 10.1002/cyto.a.20811