Endo arabinanase

Extrusion trials were performed using commercial chokeberry (Aronia melanocarpa) pomace powder (CPP) (Aronia Original Naturprodukte GmbH, Dresden, Germany) [20 (link)]. Chemicals and reagents of analytical purity grade were obtained from Merck KGaA (Darmstadt, Germany), unless stated otherwise. Amyloglucosidase (E-AMGDFPD, from Aspergillus niger, 36,000 U/g), α-amylase (E-PANAA, from pig pancreas 100,000 U/g), protease (E-BSPRPD from Bacillus licheniformis 9000 U/g), Celite 545 and endo-arabinanase (EC 3.2.1.99, from A. niger, 9 U/mg) were from Megazyme (Bray, Ireland) and Amberlite FPA53 and Ambersep 200 from Rohm and Haas Europe (Frankfurt, Germany). Thermostable α-amylase (Thermamyl 120 L, EC 3.2.1.1, from B. licheniformis, 120 KNU/g), protease (Alcalase 2.5 L, EC 3.4.21.62, from B. licheniformis, 2.5 AU/g), and Amyloglucosidase (AMG 300 L, EC 3.2.1.3, from A. niger, 300 AGU/g) were a gift from Novozymes, (Bagsværd, Denmark). Rotihydroquant C5 and D used for Karl Fischer titration as well as cyanidin-3-O-glucoside (≥96%), cyanidin chloride (≥97%), 5-caffeoylquinic acid (≥97%), quercetin-3-O-glucoside (≥99%), and quercetin dihydrate (≥99%) used as PP standards were obtained from Carl Roth GmbH & Co. KG (Karlsruhe, Germany). Ultrapure water was used for all experiments.

Standards used for quantitative analysis: methanol (≥99.9%, Carlo Erba, Val-de-Reuil, France) and acetic acid (≥99.8%, Sigma-Aldrich), inositol (99.5%, Sigma-Aldrich), D-(+)-galacturonic acid (>97.0%, Fluka BioChemika, Buchs, Switzerland).
Enzymes used for immunolabeling: galactanase (E-EGALN, lot 101001b), endo-arabinanase (E-EARAB, lot 111201c) and endoglucanase (E-CELTR, lot 110901a) (Megazyme). Pectin lyase was purified from the Peclyve commercial mixture [28 (link)].

If not stated otherwise, all chemicals used were of “p.a.” grade or better and were purchased from VWR (Radnor, PA, United States), Sigma-Aldrich (Schnelldorf, Germany), or Carl Roth (Karlsruhe, Germany). Thermostable α-amylase (EC 3.2.1.1, from Bacillus licheniformis, 20,000–60,000 U/mL) was purchased from Sigma. Protease (EC 3.4.21.14, subtilisin A from B. licheniformis, 300 U/mL), amyloglucosidase (EC 3.2.1.3, from Aspergillus niger, 200 U/mL), endo-arabinanase (EC 3.2.1.99, from A. niger, 9 U/mg), and endo-galactanase (EC 3.2.1.89, from A. niger, 408 U/mg) were purchased from Megazyme (Bray, Ireland).