The assay was performed as described by Tian et al. (2021) (link), with some modifications. In brief, 500 µL of protein solution containing 30 μg/mL E. coli-produced BdLM1 protein was incubated with 5 mg chitin, chitosan, cellulose, or xylan (Yuanye, Shanghai, China) in a 100 rpm shaker at 4°C for 6 h. The samples were centrifuged at 13,000 g for 5 min. The supernatants were collected and concentrated to a volume of approximately 100 µL. The pellets were washed three times with incubation buffer, and then resuspended in 100 µL demineralized water. Then, 50 µL of the pellet solution or the supernatant were individually incubated with 50 µL of SDS-PAGE protein loading buffer (2×; 200 mM Tris-HCl, pH 6.5, 0.4 M dithiothreitol, 8% sodium dodecyl sulfate, 6 mM bromophenol blue, and 40% glycerol) at 95°C for 10 min. Samples were analyzed with a Western blot using anti-His antibodies. Photos were taken using Azure Biosystems (Azure, Dublin, CA, USA) in a custom setting.
Xylan
Manufactured by Yuanye Bio-Technology
Sourced in China
Xylan is a plant-based polysaccharide extracted from various sources. It functions as a key structural component of plant cell walls, providing rigidity and support. Xylan is a versatile biomaterial with potential applications in the food, pharmaceutical, and industrial sectors.
Automatically generated - may contain errors
Lab products found in correlation
Chitosan, by Yuanye Bio-Technology (2 mentions)
Chitin, by Yuanye Bio-Technology (2 mentions)
Microcrystalline cellulose, by Yuanye Bio-Technology (2 mentions)
Fructo oligosaccharide, by Yuanye Bio-Technology (1 mentions)
Fructose, by Yuanye Bio-Technology (1 mentions)
Mannose, by Yuanye Bio-Technology (1 mentions)
Arabinose, by Yuanye Bio-Technology (1 mentions)
Galactose, by Yuanye Bio-Technology (1 mentions)
Rhamnose, by Yuanye Bio-Technology (1 mentions)
Xylose, by Yuanye Bio-Technology (1 mentions)
Fucose, by Yuanye Bio-Technology (1 mentions)
Sodium acetate, by Yuanye Bio-Technology (1 mentions)
P nitrophenyl β d glucopyranoside, by Yuanye Bio-Technology (1 mentions)
Lightcycler 480 2 real time pcr system, by Roche (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Yuanye Bio-Technology (1 mentions)
Xylose, by Thermo Fisher Scientific (1 mentions)
Dmso d6, by Merck Group (1 mentions)
Microcrystalline cellulose, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Tianjin Kemiou Chemical Reagent (1 mentions)
4 propylguaiacol, by Merck Group (1 mentions)
7 protocols using xylan
1
Chitin-Binding Assay for E. coli-Produced BdLM1
2
Characterization of Carbohydrates Sourced
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Glucose (Glu), Fructose (Fru), Mannose (Man), Galactose (Gal), Xylose (Xyl), Arabinose (Ara), Rhamnose (Rha), Fucose (Fuc), Galactose Uronic acid (GalUA), glucuronic acid (GlcUA), purity ≥ 98%, β-dextran (purity ≥ 80%), fructo oligosaccharide (purity ≥ 95%), xylan (purity ≥ 98%) and polygalacturonic acid (purity ≥ 90%) were purchased from Yuanye Biotechnology Co., Ltd. D-trehalose (purity ≥ 99%) was purchased from Shanghai McLean Biochemical Technology Co., Ltd. All of other chemicals and reagents were analytical grade.
3
Enzymatic Hydrolysis and Characterization
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For this study, the following instruments were purchased: an LGJ-12 vacuum freeze-dryer from Songyuan Huaxing Technology Develop Co., Ltd. (Beijing, China), an SP-756P spectrophotometer from Shanghai Spectrum Instrument Co., Ltd. (Shanghai, China) and a LightCycler 480 II Real-Time PCR System from Roche Group (Basel, Switzerland). In addition, an AWL-1002-M Aquapro ultrapure water machine (Aquapro, United States) was also used to produce the ultrapure water used in the experiments.
Glucose (cas#: 50–99-7), carboxymethylcellulose (cas#: 9000-11-7), avicel (cas#: 9004-34-6), xylan (cas#: 9014-63-5), dinitrosalicylic acid (DNS, cas#: 609–99-4), p-nitrophenyl-β-D-glucopyranoside (cas#: 2492-87-7), 2,6-dimethoxyphenol (DMP, cas#: 91–10-1), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS, cas#: 30931–67-0) and sodium acetate (cas#: 127–09-3) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China).
Glucose (cas#: 50–99-7), carboxymethylcellulose (cas#: 9000-11-7), avicel (cas#: 9004-34-6), xylan (cas#: 9014-63-5), dinitrosalicylic acid (DNS, cas#: 609–99-4), p-nitrophenyl-β-D-glucopyranoside (cas#: 2492-87-7), 2,6-dimethoxyphenol (DMP, cas#: 91–10-1), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS, cas#: 30931–67-0) and sodium acetate (cas#: 127–09-3) were purchased from Shanghai yuanye Bio-Technology Co., Ltd. (Shanghai, China).
4
Feedstock Characterization and Gas Analysis
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Wheat straw and corncob were obtained from Anhui Province, China. Pine, birch and poplar were obtained from Liaoning Province, China. And beech was provided by Dansk Træmeland. The all feedstocks were dried at 110 °C for 24 h and milled into powder with the size <40 meshes before use. Commercial nickel and nickel-aluminum powders were purchased from Dalian Tongyong Chemical Co., Ltd. MEthanol (>99%) was purchased from TCI. Ethanol (>99%) and glucose (AR) were purchased from Tianjin Kemiou Chemical Reagent Co., Ltd. DMSO-d6 (>99.9% D) and 4-propylguaiacol were purchased from Sigma-Aldrich. Microcrystalline cellulose and xylose were purchased from Alfa Aesar. Xylan was purchased from Shanghai Yuanye biotechnology Co., Ltd. Methane (>99%), ethane (>99%), propane (>99%), butane (>99%), hydrogen (>99%), carbon dioxide (>99%), and the mixed standard gas were provided by Dalian GuangMing Special Gas Products Co., Ltd. Milli-Q water was used in all the experiments.
5
Preparation of Composite Nanofibers from Cellulose, Lignin, and Xylan
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CNFs were purchased from Intelligent Chemicals
Pty Ltd (Guangzhou, China). Their average fiber length and fiber diameter
were 4–6 μm and 10–50 nm, respectively. Lignin
(Mn: 1700 g/mol) was recovered from soda
pulping effluent of the State Key Laboratory of Pulp & Paper Engineering
(Guangzhou, China). The functional groups attached to lignin benzene
rings and side chain include −OCH3 (16.0%), −C=O,
COOH (7.2%), OH alcohol, OH phenolic (2.6%), and so on. This lignin
was hydrophobic and was well dispersed in sodium hydroxide (NaOH)
solution. Xylan (Mw: 4.9 × 104 g/mol) isolated from bagasse was purchased from Shanghai
Yuanye Biotechnology Co., Ltd. (Shanghai, China). The sugar composition
was as follows: 87.35% xylose, 9.28% arabinose, 0.81% glucose, 0.50%
galactose, and 2.06% glucuronic acid. This Xylan was hydrophobic and
well dispersed in NaOH solution. Corn starch (Mw: 2.7 × 107 g/mol) without any chemical treatment
was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai,
China). The amylose/amylopectin ratio of starch was 1:3. This starch
was hydrophilic and was well dispersed after cooking. Sodium hydroxide
(NaOH), hydrogen peroxide (H2O2), ammonia (25–28%),
copper sulfate (CuSO4·5H2O), and sodium
sulfide nonahydrate (Na2S·9H2O) were all
of analytical grade.
Pty Ltd (Guangzhou, China). Their average fiber length and fiber diameter
were 4–6 μm and 10–50 nm, respectively. Lignin
(Mn: 1700 g/mol) was recovered from soda
pulping effluent of the State Key Laboratory of Pulp & Paper Engineering
(Guangzhou, China). The functional groups attached to lignin benzene
rings and side chain include −OCH3 (16.0%), −C=O,
COOH (7.2%), OH alcohol, OH phenolic (2.6%), and so on. This lignin
was hydrophobic and was well dispersed in sodium hydroxide (NaOH)
solution. Xylan (Mw: 4.9 × 104 g/mol) isolated from bagasse was purchased from Shanghai
Yuanye Biotechnology Co., Ltd. (Shanghai, China). The sugar composition
was as follows: 87.35% xylose, 9.28% arabinose, 0.81% glucose, 0.50%
galactose, and 2.06% glucuronic acid. This Xylan was hydrophobic and
well dispersed in NaOH solution. Corn starch (Mw: 2.7 × 107 g/mol) without any chemical treatment
was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai,
China). The amylose/amylopectin ratio of starch was 1:3. This starch
was hydrophilic and was well dispersed after cooking. Sodium hydroxide
(NaOH), hydrogen peroxide (H2O2), ammonia (25–28%),
copper sulfate (CuSO4·5H2O), and sodium
sulfide nonahydrate (Na2S·9H2O) were all
of analytical grade.
6
Enzymatic Deconstruction of Chitin Oligomers
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T. gamsii R1 was isolated from shrimp shell waste and identified via ITS sequencing and then conserved in our laboratory. E. coli Dh5α competent cells were purchased from Tiangen Biotechnology (Beijing, China). PrimeSTAR® HS (Premix) DNA polymerase and PrimeScript™ RT-PCR Kits were purchased from Takara Biotechnology (Beijing, China). COSs with different degrees of polymerization (DPs), including N-acetyl-D-glucosamine (DP1), N-acetyl-chitobiose (DP2), N-acetyl-chitotriose (DP3), N-acetyl-chitotetraose (DP4), and N-acetyl-chitopentaose (DP5), were purchased from Qingdao BZ Oligo Biotech (Qingdao, China). Furthermore, GlcNAc, (GlcNAc)2, (GlcNAc)3, (GlcNAc)4, and (GlcNAc)5 represent N-acetyl-D-glucosamine, N-acetyl-chitobiose, N-acetyl-chitotriose, N-acetyl-chitotetraose, and N-acetyl-chitopentaose, respectively. Powdery chitin, xylan, microcrystalline cellulose, and chitosan with a degree of deacetylation (DDA) of 85 to 95% were purchased from Yuanye Biotechnology (Shanghai, China). The colloidal and ball milled-chitin were prepared according to a previously described method [51 (link)].
7
Enzyme-Assisted Biomass Valorization
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The PrimeSTAR® HS (Premix), Ex Taq® DNA polymerase and T4 ligase were purchased from Takara Biotechnology (Dalian, China). Substrates, such as chitin, xylan, microcrystalline cellulose, soluble starch, and powdery chitosan with 85%, 90% and 95% degrees of deacetylation (DA), were from Yuanye Biotechnology (Shanghai, China). Chitosan oligosaccharides with different degree of polymerization (DP 1–6) were purchased from Qingdao BZ Oligo Biotech (Qingdao, China). Non-GMO soybeans (protein content 38%) were originally produced in Canada and supplied by Jinzai Food Group Co., Ltd. (Yueyang, China). All other chemical reagents were of analytical grade and purchased from the Guangzhou Chemical Reagent Co., Ltd. (Guangzhou, China).
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