Bbl middlebrook broth

We used the following bacterial species: Streptococcus agalactiae ATCC 12403, Staphylococcus aureus MRSA ATCC 43300, Klebsiella pneumoniae Extended Spectrum β-Lactamase (ESBL) ATCC 700603, Acinetobacter baumanii ATCC19606, and Enterococcus faecium (VRE) DSM17050. Bacteria were cultured at 37 °C/5% CO₂ in liquid THY broth (Todd-Hewitt Broth, Oxoid, Hampshire, UK) supplemented with 0.5% yeast extract (BD, Miami, FA, USA). Pseudomonas aeruginosa PA14 (DSM 19882) was cultivated in liquid Müller–Hinton–Broth (Carl Roth, Karlsruhe, Germany) at 37 °C, under orbital shaking at 160 rpm.
In addition, the employed Mycobacterium tuberculosis ATCC 27294 (Mtb) was grown in 7H9-medium containing 7H9 BBL Middlebrook broth (BD), glycerol (Honeywell, Charlotte, NC, USA) OADC (Oelic Albumin Dextrose Catalase; BD), Tween 80 (Roth, Karlsruhe, Germany), and ddH₂O. The pH was adjusted between 7.2–7.4 and sterile filtration was performed with a 0.2-µm filter membrane (Thermo Scientific^TM Nalgene^TM Rapid-Flow^TM, Thermo Scientific, Bremen, Germany).

Four bacterial species and one yeast species were used to evaluate the antimicrobial activity of the putative AMPs. Pseudomonas aeruginosa ATCC 29213, Listeria monocytogenes ATCC BAA-679/EGD-e and Klebsiella pneumoniae ATCC 70,063 were cultured at 37 °C in a 5% CO₂ atmosphere overnight as preculture inoculum in Todd–Hewitt broth. Yeast Candida albicans (ATCC 90028) was grown in Sabouraud dextrose medium at 37 °C for 48 h as preculture inoculum. Mycobacterium tuberculosis ATCC 27,294 was grown in 7H9 medium containing 7H9 BBL Middlebrook broth (BD), glycerol, OADC (Oelic Albumin Dextrose Catalase; BD), Tween 80 and ddH₂O. The pH was adjusted between 7.2–7.4, and sterile filtration was performed with a 0.2 µm filter membrane (Thermo ScientificTM NalgeneTM Rapid-FlowTM; Thermo Fisher Scientific GmbH, Dreieich, Germany).