Cellic ctec2 enzymatic cocktail

Wheat Bran (WB) was provided by Chamtor, Bazancourt, France. For the preparation of DWB, WB was washed with water at 40°C during 10 min (solid/liquid ratio: 1/10) and filtrate through a glass filter (40–100 μm pososity). Four successive washings were performed to obtain a total destarching. [C2mim][OAc] were purchased from Solvionic S.A. (Verniole, France). Standards (arabinose, galactose, glucose, xylose, cellobiose, and cellotriose) were purchased from Sigma Aldrich (Steinheim, Germany). All other chemical reagents were purchased from commercial sources in France and used as received. Cellic CTec2 enzymatic cocktail including both cellulolytic and hemicellulolytic activities was supplied by Novozymes (Bagsvaerd, Denmark) and prepared at 15 FPU/g of WB or DWB for the production of hydrolysates rich in glucose and xylose. The value of 2.0 mg of reducing sugar as glucose from 50 mg of filter paper (4% conversion) in 60 min has been designated as the intercept for calculating filter paper cellulase units (FPU) by IUPAC.

Orange waste used for laccase production was donated by a local restaurant and subsequently dried and milled. Cellic CTec2 enzymatic cocktail and commercial laccase (Novozymes NS-22127) were kindly donated by Novozymes^® (Bagsvaerd, Denmark). The chromatography columns Hiprep Q FF and Superdex 75 10/300 GL were acquired from GE Healthcare Life Science (Chicago, IL, USA). Aminex HPX-87P column and Precision Plus Protein TM Standards were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Malt Extract Agar, Vanillin, 3,5-dinitrosalicylic acid (DNS), sodium carbonate, and the substrates for enzymatic activities, 3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,6-dimethoxyphenol (DMP), xylan beechwood, carboxymethylcellulose (CMC), locust bean, debranched arabinan, β-glucan, p-nitrophenyl-α-L-arabinofuranoside, p-nitrophenyl-β-d-galactopyranoside, p-nitrophenyl-β-d-glycopyranoside, p-nitrophenyl-β-D-xylanopyranoside, and p-nitrophenyl-β-d-cellobioside, were purchased from Sigma-Aldrich (St. Louis, MO, USA). All reagents used for the assays were of analytical grade.