Shearzyme plus 2x

Enzymatic extractions were performed using seven commercial glucanases: Ultraflo XL, Ultraflo Max, Ultimase BWL 40, Viscozyme L, Celluclast 1.5L, Pectinex Ultra Tropical and Shearzyme Plus 2X (Novozymes, Bagsvaerd, Denmark). Enzymatic treatments (100 mL) were performed in water at a solid-to-solvent ratio of 1:20 (w:v) in accordance with previous studies [11 (link)]. Reaction mixtures consisted of a 1% enzyme-to-lentil hull ratio (w:w) according to the manufacturer’s recommendations and were processed at 40 °C for 3 h in a Thermomixer C shaker (Eppendorf Ibérica, Madrid, Spain) at 2000 rpm. The reaction with Pectinex^® Ultra Tropical was monitored at selected times (0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5 and 4 h) by taking 2 mL aliquots. Enzymes were deactivated at 95 °C in a water bath for 5 min.

Hulls from an industrial de-hulling process that produces red football and split lentils were kindly provided by Prairie Pulse Inc. (Vanscoy, Saskatchewan, SK, Canada) in May 2019. Hulls were ground into a fine powder with a mixer mill (MM 400, Retsch, Haan, Germany) for approximately 3 min at maximum speed and stored in sealed plastic bags at 4 °C. Ultraflo XL, Ultraflo Max, Ultimase BWL 40, Viscozyme L, Celluclast 1.5 L, Pectinex^® Ultra Tropical and Shearzyme Plus 2X were obtained from Novozymes (Bagsvaerd, Denmark). Fast Blue BB (FBBB) [4-benzoylamino-2,5-dimethoxybenzenediazonium] chloride hemi-(zinc chloride), 2,2′-azinobis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2′-diazobis-(2-aminodinopropane)-di-hydrochloride (AAPH) and fluorescein were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Standards such as (+)-catechin, trans-p-coumaric acid, quercetin 3-O-rutinoside, quercetin 3-O-glucoside and kaempferol 3-O-rutinoside were provided by Extrasynthese (Lyon, Genay Cedex, France). Standards of 6-hydroxy-2,5,7,8-tetramethyl-2-carboxylic acid (Trolox), D-glucose, glycerol and gallic acid were acquired from Sigma-Aldrich Co. (St. Louis, MO, USA).