After 18 h of incubation at 37AdegC, Akkermansia municiphila cells reached maximum OD600
of 2.10+-0.07 with oxyrase and OD600
of 1.66+-0.05 with Cys-HCl.
aeruginosa FRD1 cells (OD600
= 0.1) were inoculated into Jensen's chemically defined media and added to a sterile 24-well plate containing glass coverslips (Costar, USA).
After the samples were pretreated, the remaining concentration of estradiol in the medium was measured by high-performance liquid chromatography, and the cell density was measured at OD600
using a microplate reader.
The optical density (OD600
) was measured every 3 hours from zero point until 48 hours.
The optical density at 420 nm (OD420) was measured, and [beta]-galactosidase activity was normalized to the amount of cells estimated from the OD600
Cultures were then normalized for OD600
, serially diluted, and spotted onto synthetic solid media containing glucose or galactose lacking uracil and were grown at 30[degrees]C.
Percent (%) CSH was determined by using the following equation: [1 - (OD600
nm after vortexing/ OD600nm beforevortexing)] x 100.
Cells were allowed to grow in LB broth till OD600
aureus RN 7044 was cultured with aeration in MHB at 37[degrees] C up to OD600
A monoclonal strain that was picked from plate G418 was inoculated into BMGY medium [BMGY: 20 g tryptone, 10 g yeast extract, 100 mL 10 x yeast nitrogen base (YNB W/O amino acids), 2 mL 500 x D-Biotin (B), 100 mL 10 x glycerol (Gl), 100 mL 1 M phosphate buffer, 700 mL [H.sub.2]O], and cultured for 2 days (OD600
= 2-6) in the shaker.
values of the samples were measured by a spectrophotometer and samples that had OD600
values between 0.2 and 0.3 were used for spheroplasting.
Samples having an OD600
nm value of more than 0.7 were diluted with the sterile culture medium, and the corrected OD value was obtained by multiplying the dilution factor.