Absolute ethanol

Tetraethylortosilicate (TEOS, 99%) and Chloride acid (HCl) (37%) were obtained from Alfa Aesar (Haverhill, MA, USA). Chitosan (CS; 50,000–190,000 Da; 75%–85% deacetylation degree) and 3-glycidoxypropyltrimethoxysilane (GPTMS, >98%) were purchased for Sigma Aldrich (St. Louis, MO, USA). Absolute ethanol (99.5%) was purchased from Panreac (Barcelona, Spain). Acid acetic (Reagent Grade) was purchased for Scharlau (Barcelona, Spain). HOB^® human osteoblasts, foetal calf serum and Osteoblast Growing Medium (Promocell, Heidelberg, Germany). Paraformaldehyde, PBS, Triton x-100, bovine serum albumin, Metanol, rhodamine phalloidin and monoclonal anti-vinculin FITC conjugate were all purchased from Sigma, (St. Louis, MI, USA), Vectashield^® (Vector, Burlingame, CA, USA).

All of the amines and 2-ME were of analytical grade and obtained from Sigma (Maribor, Slovenia). NaHS was of technical grade and obtained from MOLEKULA (Ljubljana, Slovenia). Absolute ethanol was obtained from Panreac (Ljubljana, Slovenia). All of the chemicals for synthesis were obtained from Aldrich (Vienna, Austria).

Chitosan (CS; 50,000–190,000 Da; 75–85% deacetylation degree) was provided by Sigma Aldrich (St. Louis, MI, USA). Tetraethyl-orthosilicate (TEOS, 99%) and hydrochloric acid (37%, Pharma grade) were supplied for Alfa Aesar (Haverhill, MA, USA). Tri-calcium phosphate (TCP, pure, pharma grade) and absolute ethanol (99.5%) were obtained from Panreac (Barcelona, Spain), HOB^® human osteoblasts, fetal calf serum, and osteoblast growing medium (Promocell, Heidelberg, Germany) paraformaldehyde, PBS, Triton x-100, bovine serum albumin, methanol, rhodamine phalloidin, and monoclonal anti-vinculin FITC conjugate were all purchased from Sigma Aldrich, (St. Louis, MI, USA) along with Hard Set Vectashield with DAPI ^® (Vector, Burlingame, CA, USA).

Sodium hydroxide and L-ascorbic acid were from Carlo Erba (Milano, Italy). Ferulic acid was purchased from Sigma-Aldrich (Steinheim, Germany). 2,4,6-Tris(2-pyridyl)-s-triazine (TPTZ) was from Fluka (Steinheim, Germany). Sulfuric acid (96%) and sodium carbonate anhydrous were from Penta (Prague, Czech Republic). The stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) was from Alfa Aesar (Karlsruhe, Germany). Anhydrous citric acid and iron chloride hexahydrate (FeCl₃•6H₂O) were from Merck (Darmstadt, Germany). Folin-Ciocalteu reagent and absolute ethanol were from Panreac (Barcelona, Spain). All solvents used for chromatographic determinations were of appropriate (HPLC) grade.

Three components representative of the E. coli cell wall were tested as conditioning agents: mannose, myristic acid and palmitic acid. Mannose is a sugar monomer which is typically found on the walls of bacterial cells [28] . Furthermore, it is one of the predominant monosaccharide components detected on the cell surfaces of various enteropathogenic E. coli serotypes [29] (link). The two saturated fatty acids palmitic acid (C16:0) and myristic acid (C14:0) are the dominant ones in the E. coli cell wall [30] (link),[31] (link), consisting of more than 50% of the fatty acid content in continuous cultures [31] (link).
D-(+)-mannose (Fluka Analytical, cat. no. 63580, USA) was prepared in sterile distilled water at concentrations of 0.5, 1, 5, 10 and 100 g l⁻¹. Given the low solubility of the palmitic (Merck KGaA, cat. no. 800508, Germany) and myristic acid (Fluka Analytical, cat. no. 70082, USA) in water, concentrated solutions (25 g l⁻¹) were prepared using absolute ethanol (PanReac AppliChem, Germany) from which working solutions of 2.5 × 10⁻⁴, 2.5 × 10⁻³, 0.025, 0.25 and 2.5 g l⁻¹ (below the micellar concentration) were prepared in distilled water. The pH of myristic acid solutions was 6.58 ± 0.16, while the pH of palmitic acid solutions was 6.68 ± 0.12.

In this study, Comercializadora Tint Café S.A.S, Villeta, Colombia, supplied coffee pulp samples of both colors (red and yellow). β-Carotene of 97.0% (w/w) purity was used from Sigma-Aldrich (product number 7235-40-7). The chemicals 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and anhydrous caffeine (C₈H₁₀N₄O₂), were also used from Sigma-Aldrich. Absolute ethanol (C₂H₅OH, 99.8% v/v), ethyl acetate (C₄H₈O₂, 99.5% v/v), acetone (C₃H₆O, 99.5% v/v), hydrochloric acid (HCl, 37.0% v/v), and sodium hydroxide (NaOH, 98.0% w/w) were obtained from Panreac (Gmbh). For the determination of total dietary fiber (TDF), insoluble dietary fiber (IDF), and soluble dietary fiber (SDF) heat-stable α-amylase, protease, and amyloglycosidase (Sigma® Life Science) were utilized for enzymatic digestion. Anhydrous disodium hydrogen phosphate (Na₂HPO₄, 99.0% w/w) and anhydrous sodium dihydrogen phosphate (NaH₂PO₄, 99.0% w/w) from Panreac were used for preparing the phosphate buffer in TDF, IDF, and SDF quantification. For the extract dilution a Don Olio (Colombia) sunflower oil was used. As wall materials, arabic gum from CIACOMEQ S.A.S. and maltodextrin from Cimpa S.A.S. were used.

DNA extraction of the field isolates was achieved by QuickGene‐810 (AutoGen, Japan) using the QuickGene DNA tissue kit S (DT‐S), which was applied according to the manufacturer's recommendations. Briefly, 3–5 fresh colonies of each sample grown on soybean casein digest agar were transferred into a sterilized microcentrifuge tube containing 180 µl MDT lysis buffer and 20 µl proteinase K, and the lysate was then centrifuged at 8,000 g for 5 min. The supernatant was transferred to a new Eppendorf tube, and 180 µl LDT buffer was added. Two hundred forty microliters of absolute ethanol (Panreac, Barcelona, Spain) was added, and the tube was properly agitated. The lysate was transferred into the cartridge supplied with the kits and then inserted into the machine. Finally, the concentration and purity of the extracted DNA were determined by the NanoDrop™ 2000 spectrophotometer (Thermo Scientific, MA, USA).