media was replaced with media containing compounds.
This media containing compounds (500 µL) was then added to the cells.
aCV-1 cells were infected with the WR strain at 2.5 pfu/cell and then media containing compounds at 50 or 25 µg/mL were added; at 16 h post-infection, the cells and media were collected for virus titration by plaque assay.
Transposon mutagenesis libraries of E. coli and Shewanella oneidensis (more than 8,000 mutants) were grown in media containing compounds of interest, and a shortened LC-MS protocol (2 min per sample) was then used to analyse the supernatant from spent cultures.
These results were somewhat surprising, since it is not clear that the media contained compounds that inhibit T. reesei.
Spent media was removed, and fresh media added containing compound at 100 μL/well (9 doses, 3-fold dilution, 0.1% DMSO) in duplicate or triplicate.
To evaluate the ability of unnatural d-alanine derivatives to access the cell wall, we first grew bacteria in media containing R-propargylglycine (compound 1, Figure 1B, abbreviated alkDala) or R-2-amino-3-azidopropanoic acid (compound 2, Figure 1B, abbreviated azDala) for one or more generations.
After 1 hour incubation with the media containing tetrazolium compound, the absorbance at 450 nm was detected for 0.1 sec with Arvo MX 1420 (Perkin Elmer, Waltham, MA, USA).
After 7 d, the media was exchanged for fresh MS media containing the compound of interest.
