The
isolated strain E. bolteae CEBAS S4A9
and representative strains of the closest relatives (E. bolteae DSM 29485, DSM 15670T, Enterocloster asparagiformis DSM 15981T, Enterocloster citroniae DSM 19261T, and Enterocloster clostridioformis DSM 933T) obtained from the DSMZ culture collection were
used to investigate their capacity to produce final Uros in the presence
of EA and other Uro intermediaries. Briefly, isolated and DSMZ strains
were separately incubated on a WAM agar plate for 6 days. A single
colony was cultivated in a 5 mL WAM tube. Diluted inoculum (2 mL)
was transferred to WAM (20 mL), obtaining an initial load of 104 CFU mL–1. EA, Uro-M6, Uro-D, Uro-C, Uro-A,
IsoUro-A, and Uro-B were dissolved in propylene glycol and added to
the 20 mL cultures to obtain a final concentration of 15 μM
each. After incubation in an anoxic environment at 37 °C, aliquots
(5 mL) were taken periodically for high-performance liquid chromatography
(HPLC) analyses as described below.
isolated strain E. bolteae CEBAS S4A9
and representative strains of the closest relatives (E. bolteae DSM 29485, DSM 15670T, Enterocloster asparagiformis DSM 15981T, Enterocloster citroniae DSM 19261T, and Enterocloster clostridioformis DSM 933T) obtained from the DSMZ culture collection were
used to investigate their capacity to produce final Uros in the presence
of EA and other Uro intermediaries. Briefly, isolated and DSMZ strains
were separately incubated on a WAM agar plate for 6 days. A single
colony was cultivated in a 5 mL WAM tube. Diluted inoculum (2 mL)
was transferred to WAM (20 mL), obtaining an initial load of 104 CFU mL–1. EA, Uro-M6, Uro-D, Uro-C, Uro-A,
IsoUro-A, and Uro-B were dissolved in propylene glycol and added to
the 20 mL cultures to obtain a final concentration of 15 μM
each. After incubation in an anoxic environment at 37 °C, aliquots
(5 mL) were taken periodically for high-performance liquid chromatography
(HPLC) analyses as described below.
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