Yeast extract

Halomonas sp. SF2003 is cultivated in Zobell medium (Bacto Tryptone, (Difco, BD, Göteborg, Sweden) 4 g/L, Yeast Extract (Fisher BioReagents, Pittsburgh, PA, USA) 1 g/L, sea salts (Aquarium systems, Instant Ocean, Blacksburg, VA, USA) 30 g/L, pH 7.5), with an orbital agitation of 200 rpm, at 30 °C. The medium is complemented with glucose (Labogros), at 10 g/L, for pre-cultures dedicated to PHA productions.
Cupriavidus necator H16, C. necator PHB¯4, PHB¯4/pBBR1-Pro_Cn-phaC1 and PHB¯4/pBBR1-Pro_Cn-phaC2 are cultivated in nutrient-rich medium (NR medium) (Meat extract (Biokar Diagnostics, Allonne, France) 10 g/L, Yeast Extract (Fisher BioReagents, USA) 2 g/L, Peptone from gelatin, enzymatic digest (Sigma-Aldrich, St. Louis, MO, USA) 10 g/L, pH (7)), with an orbital agitation of 200 rpm, at 30 °C.
The transformant strains PHB¯4/pBBR1-Pro_Cn-phaC1 and PHB¯4/pBBR1-Pro_Cn-phaC2 were selected on Simmons citrate agar plates (Thermo Scientific™, Illkirch–Graffenstaden, France), prepared following the manufacturer instructions. For the transformant strains PHB¯4/pBBR1-Pro_Cn-phaC1 and PHB¯4/pBBR1-Pro_Cn-phaC2, the media were complemented with kanamycin (Km, Gibco, Waltham, MA, USA), at 50 µg/mL.

Wildtype strain SC5314 Candida albicans was purchased from the American Type Culture Collection (American Type Culture Collection, Manassas, VA, USA). SC5314 constitutively expressing far-red fluorescent protein (C. albicans iRFP) was kindly donated by Robert Wheeler (University of Maine, Orono, ME) [33 (link)]. C. albicans was grown in YPD liquid media (yeast extract, peptone, dextrose) containing 1% yeast extract (Acros Organics, Fair Lawn, NJ, USA), 2% peptone (BD Biosciences, San Jose, CA, USA), and 2% dextrose (Sigma-Aldrich). C. albicans was cultured overnight at 30 °C on a rotating culture wheel (Thermo Fisher Scientific). The following day, C. albicans was removed from the wheel, washed twice with PBS and resuspended in PBS. C. albicans was counted using the LUNA automatic cell counter and kept on ice until the time of the assay.

Brucella abortus 2308 is a CO₂-independent, virulent, smooth strain. The B. abortus 2308 mCherry strain constitutively expresses fluorescent mCherry due to the integration of a plasmid containing the mCherry coding sequence and a kanamycin resistance marker^{95 (link)}. Cultures of Brucella were freshly inoculated from frozen stock into yeast extract and tryptone (2YT) medium [1 % yeast extract (Invitrogen, Carlsbad, CA, USA), 1.6 % bactotryptone (Invitrogen), 0.5 % NaCl (Invitrogen)) plates (supplemented with 10 µg/ml kanamycin (AppliChem Panreac) for the mCherry strain], before subcultures were grown in 2YT broth (aerobic condition, 37 °C).
All Brucella were handled under BSL-3 containment according to the Council Directive 98/81/EC of 26 October 1998, adopted by the Walloon Government (4 July 2002).

C. auris isolate 12 (NCPF 8973), derived from the South Asian/Indian lineage, was obtained from the National Mycology Reference Laboratory (United Kingdom) (21 (link)). The strain was maintained on yeast extract–peptone–dextrose (YPD) solid medium [10 g/L of yeast extract (Alfa Aesar, USA), 20 g/L of mycological peptone (Oxoid, United Kingdom), 20 g/L of dextrose, and 20 g/L of agar (VWR International LLC, Hungary), pH 5.6]. Culturing and biofilm formation were performed in RPMI-1640 (with L-glutamine and without bicarbonate, pH 7.0, and with 3-(N-morpholino) propanesulfonic acid; Merck Ltd, Budapest, Hungary). Farnesol (Merck Ltd.) was obtained as a 3-M stock solution, which was diluted to 30 mM in 100% methanol. The working concentration of farnesol (75 µM) was prepared in the RPMI-1640 medium. Drug-free RPMI-1640 controls were supplemented with 1% (vol/vol) methanol. Tyrosol [2-(4-hydroxyphenyl) ethanol] (Merck Ltd.) was prepared as a 0.1-M stock solution in sterile physiological saline. The working concentration of Tyrosol (15 mM) was prepared in RPMI-1640.

S. aureus USA500 (Diep et al., 2006 (link)) was used for construction of gene knockout and complementation strains. E. coli DC10B (Monk et al., 2012 (link)) was used for shuttle plasmid construction. Luria Broth medium was composed of 1% tryptone (Oxoid), 0.5% yeast extract (Oxoid) and 0.5% NaCl; BM (B-Medium) was composed of 1% tryptone, 0.5% yeast extract, 0.5% glucose, 0.1% K₂HPO₄ and 0.5% NaCl; BM and TSB (Tryptic soy broth, Oxoid) were used for S. aureus cultivation. Bacterial strains were inoculated in BM, and their growth rate at 37°C was monitored by measuring the OD values at 600 nm. Anhydrotetracycline (ATc) was used for induction of secY antisense RNA during gene knockout. Antibiotics were added to medium at the following concentrations: chloramphenicol, 10 μg/ml; ampicillin, 100 μg/ml, levofloxacin, 50 μg/ml.