rCbpA (0.5 µM) was incubated with 0.05–9 mM cellobiose (Sigma) in 25 mM phosphate buffer pH 6 containing 0.01% (w/v) NaN3 in a final volume of 200 µL at 37 °C during 2 min. Then 50 µL of 0.5 M sodium hydroxide was added prior to analysis by HPAEC-PAD, using the same procedure as for cellodextrin content analyses, except that various concentrations of glucose, cellobiose, and α-d -glucose-l -phosphate (Sigma) were used to identify and quantify the released sugars. Activity of rCbpA on 1 mM cellotriose, cellotetraose, and cellopentaose was assayed with 1 µM of enzyme and incubation times up to 120 min at 37 °C were used.
Cellobiose
Manufactured by Merck Group
Sourced in United States, Germany, Japan, Switzerland, Ireland, India, China, France
Cellobiose is a disaccharide composed of two glucose molecules linked by a β-1,4-glycosidic bond. It is a core component found in the structure of cellulose, the primary structural polysaccharide in plant cell walls. Cellobiose serves as a fundamental building block for various lab applications that involve the study of plant biomass and cellulose-based materials.
Automatically generated - may contain errors
Lab products found in correlation
Maltose, by Merck Group (29 mentions)
Cysteine, by Merck Group (15 mentions)
Xylose, by Merck Group (13 mentions)
Arabinose, by Merck Group (11 mentions)
Galactose, by Merck Group (10 mentions)
Lactose, by Merck Group (10 mentions)
D galactose, by Merck Group (8 mentions)
Avicel ph 101, by Merck Group (7 mentions)
D xylose, by Merck Group (7 mentions)
Mannose, by Merck Group (7 mentions)
P nitrophenyl β d glucopyranoside, by Merck Group (7 mentions)
D glucose, by Merck Group (6 mentions)
Fructose, by Merck Group (6 mentions)
D mannose, by Merck Group (5 mentions)
Acetic acid, by Merck Group (5 mentions)
Formic acid, by Merck Group (5 mentions)
Cellotriose, by Merck Group (5 mentions)
Cellotriose, by Megazyme (4 mentions)
Cellotetraose, by Megazyme (4 mentions)
Raffinose, by Merck Group (4 mentions)
121 protocols using cellobiose
1
Cellobiose Hydrolysis by rCbpA
2
Enzymatic Hydrolysis of Cellobiose by Bgl1B
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To examine the enzymatic activity of recombinant Bgl1B, 2.0 nmol protein was incubated in 1 mL of 20 mM sodium phosphate buffer (pH 6.0) containing 0.1% (w/v) cellobiose (Sigma) at 40 °C for 15 min for the enzymatic hydrolysis of cellobiose by Bgl1B. The enzymatic reaction was stopped by incubating the reaction mixture in boiling water for 1 min. The enzymatic activity was measured by quantifying the amount of glucose produced from the hydrolysis of cellobiose using high-performance liquid chromatography (HPLC). One unit (U) of Bgl1B was defined as the amount of enzyme required to produce 1 μmol of glucose per min under the enzymatic reaction conditions described above.
3
Characterization of Biomass Polysaccharides
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All chemicals used were of the highest purity
available. Aqueous solutions were prepared in deionized water with
an electrical resistivity of ≥18 MΩ cm at 25 °C.
Cellobiose (C7252) was purchased from Merck. Cello-oligosaccharides
and hemicelluloses were purchased from Megazyme (Wicklow, Ireland):
cellotriose (O-CTR-50MG), cellotetraose (O-CTE-50MG), cellopentaose
(O-CPE-20MG), β-glucan from barley (high viscosity, P-BGBH),
β-glucans from barley (low viscosity, P-BGBL), glucomannan (GM)
from konjac tubers (low viscosity, P-GLCML), xyloglucan from tamarind
seeds (P-XYGLN), galactan from lupin seeds (P-GALLU), lichenan from
icelandic moss (P-LICHN), arabinoxylan from wheat flour (medium viscosity,
P-WAXYM), mannan (1,4-β-d -Mannan, P-MANCB), xylan from
birchwood (partially acetylated, P-ACXYL), xylan from beechwood (P-XYLNBE-10G),
and curdlan (P-CURDL). Phosphoric acid-swollen cellulose (PASC) was
prepared according to a published protocol63 (link) from microcrystalline cellulose (MCC) (d = 50 μm,
11365, Avicel PH-101, Merck). Crystalline nanocellulose (CNC, d = 10–20 nm × l = 300–900
nm, NG01NC0101) was purchased from Nanografi Nanotechnology (Ankara,
Turkey). Kraft lignin (370959) and horseradish peroxidase (77332)
were purchased from Merck.
available. Aqueous solutions were prepared in deionized water with
an electrical resistivity of ≥18 MΩ cm at 25 °C.
Cellobiose (C7252) was purchased from Merck. Cello-oligosaccharides
and hemicelluloses were purchased from Megazyme (Wicklow, Ireland):
cellotriose (O-CTR-50MG), cellotetraose (O-CTE-50MG), cellopentaose
(O-CPE-20MG), β-glucan from barley (high viscosity, P-BGBH),
β-glucans from barley (low viscosity, P-BGBL), glucomannan (GM)
from konjac tubers (low viscosity, P-GLCML), xyloglucan from tamarind
seeds (P-XYGLN), galactan from lupin seeds (P-GALLU), lichenan from
icelandic moss (P-LICHN), arabinoxylan from wheat flour (medium viscosity,
P-WAXYM), mannan (1,4-β-
birchwood (partially acetylated, P-ACXYL), xylan from beechwood (P-XYLNBE-10G),
and curdlan (P-CURDL). Phosphoric acid-swollen cellulose (PASC) was
prepared according to a published protocol63 (link) from microcrystalline cellulose (MCC) (d = 50 μm,
11365, Avicel PH-101, Merck). Crystalline nanocellulose (CNC, d = 10–20 nm × l = 300–900
nm, NG01NC0101) was purchased from Nanografi Nanotechnology (Ankara,
Turkey). Kraft lignin (370959) and horseradish peroxidase (77332)
were purchased from Merck.
4
Monosaccharide and Oxidation Product Analysis
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Monosaccharides were determined with the HPAEC-PAD system as described by Moilanen et al. [29 (link)]. The cellulose oxidation products were also analyzed with an HPAEC-PAD system according to the method described by Rantanen et al. [50 (link)] for analysis of oligosaccharides. The eluents for gradient analysis of the oxidation products were A: 1 M NaAc in 100 mM NaOH and B: 100 mM NaOH. The samples were analyzed with two different gradients named gradient 1 and gradient 2 (Table 1 ). D-Gluconic acid sodium salt (Sigma, France), D-glucuronic acid (Sigma, Switzerland), and a cellooligosaccharide standard containing cellobiose, cellotriose, and cellotetraose (Merck, Germany) were used as standards.
5
Antioxidant Capacity Evaluation Protocols
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Fast Blue BB (FBBB) [4-benzoylamino-2,5-dimethoxybenzenediazonium chloride hemi-(zinc chloride) (Rebello et al., 2016 (link)), bile extract porcine, pancreatin from porcine pancreas, pepsin from porcine gastric mucosa, α-amylase from human saliva, 2,2′-azinobis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2′-diazobis-(2-aminodinopropane)-dihydrochloride (AAPH), fluorescein, 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich, Co. (St. Louis, MO, United States). Standards of chlorogenic acid, 2-coumaric acid, 2,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, vanillic acid, and vanillin were provided by Extrasynthese (Lyon, Genay Cedex, France). Standards of 6-hydroxy-2,5,7, 8-tetramethyl-2-carboxylic acid (Trolox), gallic acid, caffeic acid, ferulic acid, vitexin, D-(+)-glucose, D-(+)-galactose, D-(+)-xylose, D-(–)-arabinose, and stachyose were acquired from Sigma-Aldrich, Co. (St. Louis, MO, United States). Standards of 33-α-L-plus 23-α-L-arabinofuranosyl-xylotetraose (XA3XX/XA2XX), 23,33-di-α-L-arabinofuranosyl-xylotriose (A2, 3XX), and xylotriose were obtained from Megazyme (Wicklow, Ireland). Raffinose and cellobiose standards were supplied by Merck (Darmstadt, Germany).
6
Biomass Pretreatment Reagents Protocol
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D-(+)-Xylose (≥99%), Cellobiose (≥99%), D-(+)-glucose (≥99%), L-(+)-arabinose (≥99%), D-(+)-mannose (≥99%), D-(+)-galactose (≥99%), formic acid (≥95%), acetic acid (≥99%), levulinic acid (≥98%), 5-hydroxymethylfurfural (≥99%), 2-furaldehyde (≥99%), sulfuric acid (H2SO4) (95–97%), phosphoric acid (H₃PO₄) (99%), aluminium phosphate (AlPO₄) (≥93%), calcium phosphate (Ca₃(PO₄)₂) (96%), iron(III) phosphate (FePO₄) (96%), and sodium phosphate (NaH₂PO₄) (96%) were purchased from Merck (Darmstadt, Germany) and used without further purification.
7
Measuring Cytosolic Calcium Dynamics
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Calcium concentration in the cytosol was measured using a luminometric method described previously [32 (link)]. Arabidopsis 7-day-old seedlings expressing cytosolic aequorin (a calcium-sensitive luminescent protein) were dark-incubated for 12 h with a freshly prepared 7.5mM coelenterazine (p.j.k) and GLR inhibitors in 1.5ml Eppendorf tube. The same concentrations of inhibitors and the control were used as described in Methods, in the “Preparation of inhibitor-containing media ” section. Aequorin luminescence emitted from the seedlings was measured using a GloMax 20/20 luminometer (Promega) working with the software GLOMAX SIS v1.10.0. After incubation, each tube was placed in a tube holder of the luminometer in total darkness. The basal luminescence was recorded for 30 s. 1mM cellobiose (Merck) was used as an elicitor to induce a transient calcium wave. The calcium wave was measured for 5 min. Finally, the remaining reconstituted aequorin was discharged by adding 1 M CaCl2 in 10% ethanol. The luminescence counts obtained with the luminometer were calibrated into Ca2+ concentrations using the equation of Rentel and Knight [31 (link)].
8
Cellulose Substrates Characterization
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Cellulose substrates, viz.,
SigmaCell Types 20, 50, and 101, cellobiose, β-d -glucose,
and 1,6-anhydro-β-d -glucose (LVG) were purchased from
Sigma-Aldrich. According to the purchase label, SigmaCell Types 20
and 50 have average particle sizes of 20 and 50 μm, respectively.
On the other hand, SigmaCell Type 101 is composed of highly purified
cellulose fibers and has an average particle size of 15–18
μm. Calcium carboxylate monohydrate for the calibration of CO
and CO2 gases was also purchased from Sigma-Aldrich. Deionized
water with a conductivity of <0.01 S/m was used for calibrating
its fraction in the pyrolysis vapors.
SigmaCell Types 20, 50, and 101, cellobiose, β-
and 1,6-anhydro-β-
Sigma-Aldrich. According to the purchase label, SigmaCell Types 20
and 50 have average particle sizes of 20 and 50 μm, respectively.
On the other hand, SigmaCell Type 101 is composed of highly purified
cellulose fibers and has an average particle size of 15–18
μm. Calcium carboxylate monohydrate for the calibration of CO
and CO2 gases was also purchased from Sigma-Aldrich. Deionized
water with a conductivity of <0.01 S/m was used for calibrating
its fraction in the pyrolysis vapors.
9
Synergistic Cellulose Hydrolysis by Cellulases
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Partially hydrolyzed cellulose was hydrolyzed with individual cellulase components or their mixtures at different loadings at 50 C for 1 and 24 h, respectively. After hydrolysis, the liquid and solids were separated by centrifugation, and the residual solids were washed, dried, and weighed to estimate cellulose solubilization. Glucose and cello-oligomers in hydrolysate were measured using a Dionex IC equipped with electrochemical detector and CarboPac PA100 column and calibrated by sugar standards of glucose, cellobiose, and cellotriose (Sigma-Aldrich). DP>3 cellodextrins were estimated by subtracting glucose, cellobiose, and cellotriose from total cellulose solubilization and confirmed by post-hydrolysis with dilute acid (Sluiter et al., 2010) . First hour (initial hydrolysis rate) and 24 h glucose and cellulose oligomer release patterns of partially hydrolyzed cellulose were monitored for various loadings of single enzyme components (CBH1 and EG2) and their combinations. Sugar release during cellulose hydrolysis by single or mixed CBH1 and EG2 were calculated as a percentage of total loaded cellulose. The degree of synergism (DOS) is defined by Equation (2).
in which, Y CBH1þEG2 is the sugar released by CBH1 and EG2 mixtures and Y CBH1 and Y EG2 are the sugar released by individual CBH1 and EG2, respectively.
in which, Y CBH1þEG2 is the sugar released by CBH1 and EG2 mixtures and Y CBH1 and Y EG2 are the sugar released by individual CBH1 and EG2, respectively.
10
Cultivation and Supernatant Production of Faecalibacterium prausnitzii
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Faecalibacterium prausnitzii strain A2-165 (DSM N°17677, DSMZ collection, Braunschweig, Germany) was grown in LYBHI medium (BHI, Difco, Detroit, USA) supplemented with 0.5% yeast extract, 1 mg/ml cellobiose (Sigma-Aldrich Chemie GmbH, Buchs, Switzerland), 1 mg/ml maltose and 0.5 mg/ml cysteine (Sigma-Aldrich), at 37°C in an anaerobic chamber. F. prausnitzii supernatant (SN) was recovered by centrifugation and filtered through 0.45-μm-pore-size filters (VWR, Haasrode, Belgium) and stored at −80°C.
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