Liquid cultures of Micrococcus strains were grown for
24–48 hours at 37°C in BHI. Cultures were normalized to 0.05
optical density at A600nm (OD600) and incubated with
indicated molar concentrations of progesterone (Tocris Bioscience) and
17β-estradiol (Tocris Bioscience) or equal volume of absolute ethanol
vehicle (Sigma Aldrich), in respective culture media (see above). To test
whether bacterial isolates were capable of growth with progesterone and
17β-estradiol as the sole carbon source, bacterial growth curves were
performed in freshly prepared mineral salt media63 supplemented with
1×10−5M progesterone and
1×10−6M 17β-estradiol or equal volume of
absolute ethanol vehicle at a normalized starting OD600 of 0.1.
Bacterial cultures were then incubated in a Cytation3 spectrophometer (BioTek)
at 37°C for 35 hours, and OD600 was recorded every 15
minutes.
24–48 hours at 37°C in BHI. Cultures were normalized to 0.05
optical density at A600nm (OD600) and incubated with
indicated molar concentrations of progesterone (Tocris Bioscience) and
17β-estradiol (Tocris Bioscience) or equal volume of absolute ethanol
vehicle (Sigma Aldrich), in respective culture media (see above). To test
whether bacterial isolates were capable of growth with progesterone and
17β-estradiol as the sole carbon source, bacterial growth curves were
performed in freshly prepared mineral salt media63 supplemented with
1×10−5M progesterone and
1×10−6M 17β-estradiol or equal volume of
absolute ethanol vehicle at a normalized starting OD600 of 0.1.
Bacterial cultures were then incubated in a Cytation3 spectrophometer (BioTek)
at 37°C for 35 hours, and OD600 was recorded every 15
minutes.