Viscozyme l

The lyophilized microalgal biomass of Nannochloropsis gaditana (batch L3250520) was purchased from Cianoalgae SI (Gipuzkoa, Spain). Enzymatic solutions (Viscozyme^® L, Celluclast^® 1.5 L, Saczyme^® Yield) were kindly donated by Novozymes A/S (Bagsvaerd, Denmark). The main components of the enzymatic preparations and their activities are reported in Table 2.
Chloroform, methanol, and ethanol were purchased from Fisher Scientific GmbH (Wien, Austria). Sodium hydrogen carbonate, potassium hydroxide, Triton X, and dimethylsulfoxide (DMSO) were purchased from Carl Roth (Karlsruhe, Germany). Cell culture media and respective supplements were obtained from Gibco Thermo Fisher Scientific (Waltham, MA, USA) and Szabo Scandic (Vienna, Austria). CellTiter-Blue (CTB) 10× concentrate was purchased from Promega (Waldorf, Germany). Sulforhodamine B sodium salt (SRB) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (München, Germany). Invitrogen CyQuant LDH Cytotoxicity Assay Kit was bought from Thermo Fisher Scientific (Waltham, MA, USA). Fatty acid methyl esters standard (Supelco 37 FAME Mix) was from Supelco (Bellefonte, PA, USA).

Cellic^® CTec2 preparation (Novozymes, Franklinton, NC, USA) containing a complex of cellulolytic enzymes (75 FPU/mL activity) was used in the enzymatic hydrolysis of cellulose. The preparation was applied according to the manufacturer’s instructions at pH 5.5 and 50 °C. Selected fermentation media were also prepared using Viscozyme^® L (Novozymes, Bagsvaerd, Denmark). This preparation exhibits endo-β-glucanase (xylanase, cellulase, hemicellulase) activity, 100 FBG g-1, and hydrolyses β-(1,3)- or β-(1,4)-glycosidic linkages in β-D-glucans. It was used according to the manufacturer’s instructions at a dose of 10 FGB/g DW at pH 5.5 and 50 °C.

The dried powders of crude metalloprotease, MP, were yielded from marine bacterium Flavobacterium sp. YS-80-122. A commercial metalloprotease concentrate produced from Bacillus subtilis, DENIE-B LPS-P, was purchased from Denykem Ltd. (Shanghai, China). Cell wall degrading enzyme complex from Aspergillus sp., ViscozymeL, containing a wide range of carbohydrases and metalloprotease was obtained from Novozymes, Denmark. The 4-aminophenylboronic acid, 3-aminophenylboronic acid (3-APBA), aniline and cyanuric chloride (2,4,6-trichloro-1,3,5-triazine) were purchased from Sigma-Aldrich, St. Louis, MO, USA. Activated Sepharose 6B with two different spacer arm lengths (5-atoms, 10-atoms) were from Beijing Weishibohui Chromatography Technology Co., Beijing, China. All remaining reagents were of analytical grade (Sinopharm Chemical Reagent, Shanghai, China).

If not otherwise indicated, suppliers for chemicals were Th. Geyer (Renningen, Germany) and Sigma-Aldrich (Taufkirchen, Gemany). Ultrapure water was produced with an Elga-Veolia-Purelab (Flex2) unit, while ‘Millipore-water’ came from a Millipore-Synergy-UV-system (Type 1). Compostable bags (10 different brands) designated for the collection of organic waste by the supplier were bought from different local supermarkets (Table S1). Polymer reference materials for BDP were: PLA (batch no.: GH0728B133, commercial name: Ingeo Biopolymer 4043D, supplier: NatureWork, Minnetonka, MN) and PBAT (batch no.: 95010016KO, commercial name: Ecoflex F Blend C1200, supplier: BASF). Protease A-01 (activity: > 1.100 U mL⁻¹), Pektinase L-40 (activity: > 900 U mL⁻¹, Exo PGA, > 300 U mL⁻¹ Endo PGA, > 300 U mL⁻¹ Pektinesterase), and Cellulase TXL (activity: > 30 U mL⁻¹) were from ASA Spezialenzyme GmbH (Wolfenbüttel, Germany), Viscozyme L (activity: > 100 FBG U g⁻¹) was from Novozymes A/S (Bagsværd, Denmark).

Based on the CSE (A), olive oil pomace (1 g) was placed in 30 mL of water and the pH was adjusted to 5.0 prior to the addition of 1.0% of enzyme (cellulase or viscoenzyme; Celluclast, batch: CCN03196, Viscozyme L, batch: KTN02300, Novozymes, Bagsværd, Denmark). The extractions were performed at 50 °C and 120 rpm for 120 min (repeated twice). After, the mixtures were filtered and lyophilized to obtain a dried extract [35 (link)].

Celluclast 1.5 L, Cellic CTec 2, Viscozyme L (manufactured by Novozymes Corp.), oat-spelt xylan (95590), Amberlite XAD4, and Amberlite IRA958 were purchased from Sigma (St. Louis, MO, USA). The T. reesei β-mannanase (TrMan5A) was obtained as previously described by Hägglund et al. [73 (link)]. AcGGM from thermomechanical pulping (TMP-AcGGM) was obtained as described in Andersson et al. [40 (link)]. Mannobiose (M2), mannotriose (M3), mannotetraose (M4), mannopentaose (M5), mannohexaose (M6), galactosyl-mannotriose (GM3), di-galactosyl-mannopentaose (G2M5), para-nitrophenyl (pNP)-α-D-galactopyranoside, pNP-β-D-glucopyranoside, pNP-β-D-mannopyranoside, pNP-β-D-xylopyranoside and low viscosity locust bean gum (lvLBG) were purchased from Megazyme (Bray, Ireland). Acetone and pullulan molecular weight standards were procured from Merck (Darmstadt, Germany).

Cellic® CTec2 preparation (Novozymes, Franklinton, NC, USA) containing a complex of cellulolytic enzymes with 75 FPU (filter-paper units)/mL activity was used in the enzymatic hydrolysis of cellulose. The preparation was applied according to the manufacturer’s instructions at pH 5.5 and 50 °C. In the preparation of fermentation media, Viscozyme® L (Novozymes, Bagsvaerd, Denmark) was additionally used. This preparation exhibits endo-β-glucanase (xylanase, cellulase, hemicellulase) activity, 100 FBG/g, and hydrolyses β-(1,3)- or β-(1,4)-glycosidic linkages in β-D-glucans. It was used according to the manufacturer’s instructions at a dose of 8 FGB (beta glucanase units)/g DW, at pH 5.5 and 50 °C.