Carob galactomannan

Phosphoric acid swollen cellulose (PASC) was prepared as follows. 5 g of Avicel^® PH-101 was moistened with water and treated with 150 mL ice cold 85% phosphoric acid, stirred on an ice bath for 1 h. Then 500 mL cold acetone was added while stirring. The swollen cellulose was filtered on a glass-filter funnel and washed 3 times with 100 mL ice cold acetone and subsequently twice with 500 mL water. PASC was then suspended in 500 mL water and blended to homogeneity.
High-purity pachyman (β-d-1,3-glucan), barley β-glucan (β-d-1,3-1,4-glucan), lichenan (from Icelandic moss, β-d-1,3-1,4-glucan), mannan (borohydride reduced), konjac glucomannan (β-d-1,4), carob galactomannan, larch arabinogalactan, wheat arabinoxylan, cellotriose, cellotetraose, cellopentaose, cellohexaose, mannobiose, and xylobiose were purchased from Megazyme. Locust bean gum, carboxymethyl cellulose (CMC), beechwood xylan, and cellobiose were purchased from Sigma. 5-bromo-4-chloro-3-indolyl-β-d-cellobioside was purchased from Santa Cruz Biotechnology.

Acetylated galactoglucomannan (hereafter referred to as β-mannan) was produced in-house from steam-exploded Norway spruce, as described previously [18 (link)]. Mannopentaose, xylopentaose, cellopentaose, konjac glucomannan, carob galactomannan, and arabinogalactan from larch wood were purchased from Megazyme. Alpha-Mannan from Saccharomyces cerevisiae was purchased from Sigma. Acetylated arabinoglucuronoxylan was produced in-house following a protocol described in [19 (link)]. Glycan stocks (10 mg/mL) were prepared using purified H₂O and sterilized by filtration using a 0.22-μm syringe filter (Sarstedt AG & Co, Nümbrecht, Germany).

α-l-Arabinofuranosidase (EC 3.2.1.55), exo-β-1,3-galactanase (EC 3.2.1.145), endo-β-1,4-mannanase (EC 3.2.1.78) and α-galactosidase (EC. 3.2.1.22) were purchased from NZYtech (Lisboa, Portugal). β-Galactosidase (EC 3.2.1.23), β-glucuronidase (EC 3.2.1.31), and α-l-rhamnosidase (EC 3.2.1.40) were purchased from Megazime (Wicklow, Ireland). Larch wood arabinogalactan polysaccharide and carob galactomannan were obtained from Megazyme (Wicklow, Ireland). Deionized water was obtained from Millipore cartridge equipment (Bedford, MA). All other solvents and chemicals were obtained from Sigma Aldrich (L’lsle d’Abeau Chesnes, France).

Carboxymethyl cellulose sodium salt (CMC), Avicel (PH-101), glucose, cellobiose, salicin, locust bean gum mannan and laminarin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Konjac glucomannan (KGM), birchwood xylan, beechwood xylan, lichenan, guar galactomannan, carob galactomannan, larch arabinogalactan and wheat arabinoxylan were purchased from Megazyme (Bray, Co. Wicklow, Ireland). Whatman^® filter paper (75 mm × 100 mm) was cut into circle paper units of 0.5 mm × 0.5 mm. Kanamycin and chloramphenicol were purchased from PanReac AppliChem (Ottoweg, Darmstadt, Germany).

Konjac glucomannan and carob galactomannan were purchased from Megazyme International (Wicklow, Ireland).