Kh2po4

Polyvinyl alcohol (PVA, M_W 145,000, fully hydrolyzed), Ca(NO₃)₂·H₂O, Ca(NO₃)₂·4H₂O, NaCl, NaHCO₃, KCl, (NH₄)₂HPO₄, K₂HPO₄·3H₂O, MgCl₂·6H₂O (all Merck, Prague, Czech Republic), hyaluronic acid (HA, M_W 1,800,000, ZVC Dr. L. Hoffmann, Cítov pod Řípem, Czech Republic), CaCl₂, NaH₂PO₄·2H₂O (Penta, Praha, Czech Republic), KH₂PO₄ (Lach-Ner, Neratovice, Czech Republic). MgCl₂·6H₂O, HCl, Na₂SO₄, Tris (all Lachema, Brno, Czech Republic).

2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulphonic acid), (ABTS); 2,2-diphenyl-1-picrylhydrazyl hydrate free radical (DPPH^•, 95%); gallic acid (GA, 3,4,5-trihydroxybenzoic acid, 99%); 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, 97%); and Na₂CO₃ were purchased from Sigma-Aldrich (Steinheim, Germany). Folin–Ciocalteu’s phenol reagent (2 M); 2-(3-hydroxy-6-oxo-xanthen-9-yl)benzoic acid (fluorescein); and 2,2′-azobis-2-methyl-propanimidamide dihydrochloride (AAPH) were from Fluka Analytical (Bornem, Belgium). KCl, NaCl, K₂S₂O₈, and KH₂PO₄ were from Lach-Ner (Brno, Czech Republic). Na₂HPO₄ and isoamyl alcohol (a mixture of isomers) were from Merck KGaA (Darmstadt, Germany). Agricultural origin ethanol (96.3%) was from Stumbras (Kaunas, Lithuania). Liquid nitrogen was from AGA SIA (Riga, Latvia). CO₂ and N₂ gases (99.9%) were from Gaschema (Jonava region, Lithuania). Perchloric acid; 2-thiobarbituric acid; 1,1,3,3 tetraethoxypropane; butylated hydroxytoluene; sulfuric acid; and boric acid were purchased from Sigma-Aldrich Chemie (Steinheim, Germany). Sodium hydroxide (NaOH, 50%) was from Ingle AS (Ingliste, Estonia), and Kjeltabs FOSS Analytical A/S was from Oridor Eesti OÜ (Tartu, Estonia).

Trichoderma asperellum MSCL 309 isolate [35 (link)] was chosen from the Microbial Strain Collection of Latvia. The fungi were cultivated on malt extract agar (Biolife Italiana, Monza, Italy) plates for 7 days at room temperature, and then the plates were stored at −70 °C for further use.
Trichoderma asperellum was cultivated on four different media, both without (M1, M2, M3, M4) and with (MT1, MT2, MT3, MT4) the addition of 0.2% Tween 80 (Sigma-Aldrich, Saint Louis, MO, USA) in a bioreactor and 1% Tween 80 in flasks, with three replicates of each. The media compositions were as follows: malt extract broth (M1) (Bioefekts, Salaspils, Latvia) at 70 mL/L; yeast extract medium (M2) consisted of yeast extract (Biolife, Italy) at 3 g/L, sucrose (Sigma-Aldrich, Saint Louis, MO, USA) at 20 g/L; salt medium (M3)—sucrose at 10 g/L, KH₂PO₄ at 1 g/L, MgSO₄ at 0.5 g/L, FeSO₄ (Sigma-Aldrich, Saint Louis, MO, USA) at 0.02 g/L, NaNO₃ (Sigma-Aldrich, Saint Louis, MO, USA) at 2 g/L, and peptone bacteriological (Biolife Italiana, Monza, Italy) at 1.12 g/L; and sugar beet molasses medium (M4)—molasses (KGM, Ropaži, Latvia) at 10 g/L, KH₂PO₄ (Lach-Ner, Neratovice, Czechia) at 2 g/L and MgSO₄ (Chempur, Piekary Śląskie Poland) at 0.2 g/L. The media were sterilized for 20 min for flask cultivations and 30 min for bioreactor cultivations at a temperature of 121 °C and a pressure of 1 atmosphere.