Milli q reference

S. enterica serovar Enteritidis strain 3934 (Solano et al., 1998 ), a clinical isolate described previously, was used for HTS experiments. The clinical isolates of S. enterica serovar Enteritidis HC164372.7 and S. enterica serovar Typhimurium HC142156206 (Microbiology Laboratory at the Hospital Clinic of Barcelona, Spain) and strain S. enterica serovar Typhimurium ATCC 14028 (obtained from the American Type Culture Collection) were used to validate the effect of culture media on Salmonella biofilms.
Cultures of the different S. enterica strains were grown in LB-agar (10 g/l NaCl, 10 g/l tryptone, 5 g/l yeast extract, and 15 g/l agar), CFA (10 g/l casamino acids, 1.5 g/l yeast extract, 0.4 mM MgSO₄, and 0.4 mM MgCl₂; Suzuki et al., 2002 (link)) or E medium. E minimal medium was prepared by diluting to 1X a stock of Vogel-Bonner salts 50X (0.8 mM MgSO₄, 9.5 mM citrate, 57 mM K₂HPO₄, and 17 mM NaNH₄HPO₄; Vogel and Bonner, 1956 ) with sterile Milli-Q water (Milli-Q Reference, Millipore).

Human serum albumin (HSA) was purchased from the Japan Blood Products Organization. Hemoglobin (Hb) was purified from bovine red blood cells purchased from Tokyo Shibaura Zouki Co. Ltd. Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) was purchased from Wako Pure Chemical Industries Ltd. 1,11-Bismaleimido-triethyleneglycol (BMTEG) was purchased from Thermo Fischer Scientific K.K. The water was deionized (18.2 MΩcm) using water purification systems (Elix UV and Milli Q Reference; Millipore Corp.). Native-PAGE and SDS-PAGE were conducted using an electrophoresis power supply (EPS 301; GE Healthcare) with precast 5‒20% polyacrylamide gel (Hi-QRAS Gel N 5‒20%: Kanto Chemical Co. Inc.). Isoelectric focusing (IEF) was performed using an electrophoresis power supply (EPS 601; GE Healthcare) with a pH 3–10 IEF protein gel (Novex; Thermo Fischer Scientific K.K.). The voltage was raised gradually to 500 V for 2 h.

The chemical compounds used for the antioxidant methods, the reagents for LOX and AChE inhibition assays and reference antibacterial and antifungal compounds were purchased from Sigma-Aldrich (Madrid, Spain). All compounds were of analytical grade (purity higher than 95%). All culture media were acquired from VWR Chemicals (Barcelona, Spain): Mueller-Hinton agar (MHA), Mueller-Hinton broth (MHB), Roswell Park Memorial Institute medium (RPMI-1640), Sabouraud dextrose agar (SDA), tryptic soy broth (TSB) and yeast peptone dextrose (YPD). Solvents of analytic grade and buffers were purchased from Merck (Madrid, Spain). Type I (18 MΩ·cm) deionized water (MilliQ-Reference, Millipore, Madrid, Spain) was used in this work.

Bovine Hb was purified from fresh bovine blood purchased from Tokyo Shibaura Zouki Co. Ltd. Purification procedures of native CSA from canine plasma are presented in Supplementary Materials. All other reagents were purchased from commercial sources as special grades and were used without further purification. Deionized water (18.2 MΩ·cm) was prepared using water purification systems (Elix UV and Milli Q Reference; Millipore Corp.). SDS-PAGE and Native-PAGE were performed using an electrophoresis power supply (EPS 301; GE Healthcare UK Ltd.) and 5–20% polyacrylamide precast gradient gel (Hi-QRAS gel N; Kanto Chemical Co. Inc.). Isoelectric focusing (IEF) was conducted using an electrophoresis power supply (EPS 601; GE Healthcare UK Ltd.) and IEF protein gels (Novex pH 3–10; Thermo Fisher Scientific Inc.). The UV-visible absorption spectra were recorded using a UV-visible spectrophotometer (8543; Agilent Technologies Inc.) connected with a temperature control unit (89090A; Agilent Technologies Inc.). Circulation dichroism (CD) spectra were recorded using a spectrophotometer (J-820; Jasco Corp.). Mass spectra were obtained using a MALDI–TOF mass spectrometer (autoflex equipped with a pulsed N₂ laser (337 nm); Bruker Daltonics K.K.). As the matrix, 0.1% sinapic acid, 0.05% trifluoroacetic acid, and 50% acetonitrile solution were used.

AmB was procured from North China Pharmaceutical Huasheng Co. Ltd. (Hebei, China). Hydrogenated soy phosphatidylcholine (HSPC), distearoylphosphatidylglycerol (DSPG), cholesterol, and hydrochloric acid (37%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hanmi Gas Co. Ltd. supplied carbon dioxide with a high purity of 99.9% (Seoul, Republic of Korea). Ethyl alcohol (purity 99.9%), mEthyl alcohol (MeOH, purity 99.5%), chloroform (CHCl₃, purity 99.5%), dimethyl sulfoxide (DMSO, purity 99.5%), N,N-dimethylacetamide (DMA, purity 99.5%), and N,N-dimethylformamide (DMF, purity 99.5%) were purchased from Samchun Pure Chemical (Pyeongtaek-si, Republic of Korea). All other chemicals and reagents were of analytical grade. Purified water of Milli-Q quality (Milli-Q Reference, Millipore^®, Molshiem, France) was used throughout the study.