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.
D gluconic acid sodium salt
Manufactured by Merck Group
Sourced in United States
D-Gluconic acid sodium salt is a chemical compound used as a laboratory reagent. It is the sodium salt of D-gluconic acid, a sugar acid. The compound is typically used in various analytical and experimental procedures in scientific research and development.
Automatically generated - may contain errors
Lab products found in correlation
Cellobiose, by Merck Group (1 mentions)
D glucuronic acid, by Merck Group (1 mentions)
Orotic acid, by Merck Group (1 mentions)
Lactobionic acid, by Merck Group (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
L lactic acid, by Merck Group (1 mentions)
D galactose, by Merck Group (1 mentions)
α d lactose monohydrate, by Thermo Fisher Scientific (1 mentions)
Citric acid, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Pyruvic acid, by Merck Group (1 mentions)
Magnesium chloride hexahydrate, by Merck Group (1 mentions)
β nicotinamide adenine dinucleotide phosphate, by Merck Group (1 mentions)
Polyethylene glycol, by Merck Group (1 mentions)
Sodium phosphate dibasic na2hpo4, by Merck Group (1 mentions)
Sodium phosphate monobasic monohydrate nah2po4 h2o, by Merck Group (1 mentions)
Adenosine 5 triphosphate disodium salt, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
Ulc ms grade, by Biosolve (1 mentions)
5 protocols using d gluconic acid sodium salt
1
Monosaccharide and Oxidation Product Analysis
2
Lactose and Whey-Derived Compound Analysis
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Firstly, α-d -Lactose monohydrate (98%, Acros Organics, Fair Lawn, NJ, USA), lactulose (99%, Alfa Aesar, Haverhill, MA, USA), lactobionic acid (97%, Sigma-Aldrich, St. Louis, MO, USA), D-(+)-glucose (99%, Sigma-Aldrich), D-(+)-galactose (99%, Sigma-Aldrich), D-gluconic acid sodium salt (99%, Sigma-Aldrich), pyruvic acid (98%, Sigma-Aldrich), L-(+)-lactic acid (85%, Sigma-Aldrich), formic acid (99%, Fisher Scientific, Waltham, MA, USA), citric acid (99%, Sigma-Aldrich), and orotic acid (98%, Sigma-Aldrich) were purchased from commercial suppliers. Reduced ruthenium supported on activated carbon (5% Ru/C, Alfa Aesar) was purchased from commercial suppliers, and it was used without further preparation. Sweet whey permeate (SWP) was obtained from a regional cheese factory (Valley Queen Co., Milbank, SD, USA), while the acid whey permeate (AWP) was obtained from a Greek yogurt plant (Chobani Co., Twin Falls, ID, USA).
3
Enzymatic Assay for Gluconate Metabolism
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Aniline (C6H7N), poly(2-acrylamido-2-methyl-1-propanesulfonic acid, PAAMPSA, Mw = kD), poly(ethylene glycol), diglycidyl ether (PEDGE, Mw = 500 Da) solution, adenosine 5′-triphosphate disodium salt (ATP) (99%), β-nicotinamide–adenine dinucleotide phosphate (NADP+, >90%), β-nicotinamide-adenine dinucleotide phosphate (reduced form, NADPH, >90%), magnesium chloride hexahydrate (99%), D-gluconic acid sodium salt (99%), sodium phosphate dibasic (Na2HPO4), sodium phosphate monobasic monohydrate (NaH2PO4-H2O) and potassium chloride (KCl) have been purchased from Sigma Aldrich (St. Louis, MO, USA). 6PGDH (EC 1.1.1.44, 150 U/mL) and GK (EC 2.7.1.12, 1500 U/mL) have been purchased from CPC Biotech (CPC Biotech, Napoli, Italy).
4
Growth Characterization of Salmonella Gallinarum Strains
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SG287/91 and its derivative engineered strains were cultured in 10 mL of LB broth for 24 hours at 37 °C. Next day, the cultures were streaked onto Brilliant Green agar (BGA) without antibiotics and colony morphology was assessed. In parallel, cultures were centrifuged at 3,000 x g for 10 minutes and suspended in 1 mL sterile PBS, pH 7.4 (10-fold concentrated). A volume of the concentrated cultures was inoculated in 10 mL of M9 minimal medium so that the initial OD595 was about 0.05 for every culture. M9 medium was prepared as described by Jelsbak et al. [21 (link)] saved for its supplementation, in this study, with 0.5% tryptone due to S. Gallinarum auxotrophy. D-Gluconic acid sodium salt (Sigma-Aldrich, US) was added as the sole carbon source at a 0.5% concentration. M9 cultures were incubated at 37 °C in a shaker incubator (150 rpm) and bacterial growth was monitored at 595 nm (OD595) in an ELISA plate reader (iMark™ Microplate Absorbance Reader, Bio-RAD, US) by measurement at every hour. Three independent experiments were performed and for each of which, samples were run in duplicates. Growth curves were extracted using Excel (Microsoft, San Diego, CA) along with the exponential equations and coefficients of determination (R2) (S1 File ).
5
Quantifying GA and 2-KGA in Bacterial Cultures
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Total RNA extraction for qRT-PCR gene expression analysis was performed as described previously on 18-days-old seedlings [48] . Transcript levels were calculated relative to the reference gene At1g13320 [49] using the 2 ÀDCT method [50] .
Detection of GA and 2-KGA GA and 2-KGA concentrations in bacterial culture filtrates were determined using ultra-performance liquid chromatographymass spectrometry (UPLC-MS). Compounds were separated on a Waters Acquity UPLC BEH Amide Column (130A ˚, 1.7 mm particle size, 2.1 mm X 50 mm) by an Acquity UPLC system (Waters, Milford, MA, USA). The mobile phase A was 90% water, 10% acetonitrile, 0.1% formic acid and the mobile phase B was 100% acetonitrile, 0.1% formic acid. All solutions were ULC/MS grade from Biosolve BV (Valkenswaard, the Netherlands) The gradient was set from 10% to 90% with a flow rate of 0.25 mL min -1 . The run time was 6 min and the inject volume was 1 ml. Mass spectrometric detection was performed in negative ionization mode m/z 50 -1250 and SIR of 2 channels m/z 193 and m/z 195 on a Waters Acquity QDa detector (Waters, Milford, MA, USA). GA and 2-KGA was quantified by peak area obtained from standards for D-Gluconic acid sodium salt (Sigma-Aldrich, St. Louis, MO) and 2-Keto-D-gluconic acid hemi-calcium salt hydrate (Sigma-Aldrich, St. Louis, MO).
Detection of GA and 2-KGA GA and 2-KGA concentrations in bacterial culture filtrates were determined using ultra-performance liquid chromatographymass spectrometry (UPLC-MS). Compounds were separated on a Waters Acquity UPLC BEH Amide Column (130A ˚, 1.7 mm particle size, 2.1 mm X 50 mm) by an Acquity UPLC system (Waters, Milford, MA, USA). The mobile phase A was 90% water, 10% acetonitrile, 0.1% formic acid and the mobile phase B was 100% acetonitrile, 0.1% formic acid. All solutions were ULC/MS grade from Biosolve BV (Valkenswaard, the Netherlands) The gradient was set from 10% to 90% with a flow rate of 0.25 mL min -1 . The run time was 6 min and the inject volume was 1 ml. Mass spectrometric detection was performed in negative ionization mode m/z 50 -1250 and SIR of 2 channels m/z 193 and m/z 195 on a Waters Acquity QDa detector (Waters, Milford, MA, USA). GA and 2-KGA was quantified by peak area obtained from standards for D-Gluconic acid sodium salt (Sigma-Aldrich, St. Louis, MO) and 2-Keto-D-gluconic acid hemi-calcium salt hydrate (Sigma-Aldrich, St. Louis, MO).
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