B. theta strains were grown anaerobically (5% H2, 10% CO2, rest N2) at 37°C, overnight in brain heart infusion (BHI) supplemented media (BHIS: 37 g/L BHI [Sigma]; 1 g/L-cysteine [Sigma]; 1 mg/L Hemin [Sigma]). For enrichment cultures in plates, we used BHI-blood agar plates (37 g/L BHI [Sigma]; 1 g/L-cysteine [Sigma]; 10% v/v defribinated sheep blood [Sigma]). Antibiotics were added to liquid cultures or plates as required for strain selection: erythromycin 25 µg/L or tetracycline 2 µg/L. In the case of BHI-blood agar plates used for cloning or gut content enrichment, we additionally added gentamycin 200 µg/L to all plates to prevent growth of other microbiota species. Plates were incubated for 48–72 hr at 37°C in anaerobic conditions. For a complete list of the bacterial strains used in this study, see Key Resources Table.
Cysteine
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Switzerland, Japan, China, France, Macao
Cysteine is a non-essential amino acid used as a laboratory reagent. It is a white crystalline solid that is soluble in water and organic solvents. Cysteine plays a role in the formation of disulfide bonds in proteins and serves as a precursor to other sulfur-containing compounds.
Automatically generated - may contain errors
Lab products found in correlation
Methionine, by Merck Group (37 mentions)
Glutathione (gsh), by Merck Group (36 mentions)
Glycine, by Merck Group (27 mentions)
Histidine, by Merck Group (24 mentions)
Alanine, by Merck Group (22 mentions)
Serine, by Merck Group (22 mentions)
Glutamic acid, by Merck Group (21 mentions)
Phenylalanine, by Merck Group (20 mentions)
Valine, by Merck Group (20 mentions)
Leucine, by Merck Group (19 mentions)
Tryptophan, by Merck Group (18 mentions)
Isoleucine, by Merck Group (18 mentions)
Hemin, by Merck Group (16 mentions)
Maltose, by Merck Group (16 mentions)
Threonine, by Merck Group (15 mentions)
Papain, by Merck Group (15 mentions)
Cellobiose, by Merck Group (15 mentions)
Glutamine, by Merck Group (15 mentions)
Tyrosine, by Merck Group (14 mentions)
Asparagine, by Merck Group (14 mentions)
321 protocols using cysteine
1
Anaerobic Growth of Bacteroides theta
2
Synthesis and Purification of Manganese Porphyrins
3
Analytical Grade Chemicals Protocol
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All chemicals used in this work were of analytical grade and were used without further purification. Glycine and cysteine were purchased from Sigma Aldrich, UK. Anhydrous sodium sulphates (Na2SO4), sodium chloride (NaCl), CaCl2, NaHPO4, NaHCO3, KCl, MgCl2, were purchased from Merck, London. Potassium Chloride (KCl), were obtained from BDH. Ascorbic acid, uric acid, glutathione, albumin, cysteine, Glycine and polyethylene oxide resin were products of Sigma-Aldrich. All experimental solutions were prepared using double deionized water (MilliQ system, Millipores).
4
Biochemical Assay Materials Analysis
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Casein (from bovine milk), Coomassie brilliant blue G-250, cysteine, bovine serum albumin, Elastin-Congo Red, Hide Powder Azure, Keratin Azure, and Tris were purchased from Sigma-Aldrich (St. Louis, Missouri, USA). Ethylenediaminetetraacetic acid was purchased from Invitrogen (Carlsbad, California, USA), sodium phosphate (98%) from Carlo Erba (Rodano, MI, Italy), detergent Azymol 6SE from Pellital (Victoria, BA, Argentina). All other chemicals were of analytical grade.
5
Cultivation of Oligo-MM12 Consortium Bacteria
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A list of bacterial strains is provided in Table S4 . Bacteria were grown in an anaerobic atmosphere of 10% carbon dioxide, 5% hydrogen, and 85% nitrogen. Members of the Oligo-MM12 consortium were cultured in BHI (Becton Dickinson) supplemented with 5 μg/ml hemin (Sigma), 5 μg/ml vitamin K1 (Sigma), 250 mg/L cysteine, 250 mg/L sodium sulfide, and 4 g/L porcine mucin type 3 (Sigma) (only for Akkermansia muciniphila YL44), with the exception of Lactobacillus reuteri I49, which was grown in MRS (Becton Dickinson).
6
Simulated Infant Gut Digestion
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Sheep milk and cow milk used in this work were provided by commercial dairy farms and were frozen before use. Baby feces were donated and stored at −80 °C shortly after collection.
Chemicals used were as follows: KCl (99.5%), KH2PO4 (99.5%), MgCl2(H2O)6, orthophosphoric acid (85%), and diethyl ether (98%) were sourced from BDH Laboratory Supplies, Dorset, England. NaCl (99.6%) and CaCl2 (99%) were sourced from ThermoFisher Scientific, (Waltham, MA, USA). NaHCO3 (99%) was sourced from JT Baker, (Phillipsburg, NJ, USA). Pancreatin (37452 FIP-U/mg) was sourced from AppliChem GmbH, (Darmstadt, Germany). D2O (99.8%) was sourced from Cambridge Isotope Labs Inc, (Tewksbury, MA, USA). (NH4)2CO3 (99%), pepsin, gastric lipase (20,000 U/mg), cysteine (97%), bile salts, ethylbutyric acid (98%), N-methyl-N-t-butyldimethylsilyltrifluoroacetamide (95%), and imidazole (99%) were sourced from Sigma-Aldrich, (St. Louis, MO, USA).
Chemicals used were as follows: KCl (99.5%), KH2PO4 (99.5%), MgCl2(H2O)6, orthophosphoric acid (85%), and diethyl ether (98%) were sourced from BDH Laboratory Supplies, Dorset, England. NaCl (99.6%) and CaCl2 (99%) were sourced from ThermoFisher Scientific, (Waltham, MA, USA). NaHCO3 (99%) was sourced from JT Baker, (Phillipsburg, NJ, USA). Pancreatin (37452 FIP-U/mg) was sourced from AppliChem GmbH, (Darmstadt, Germany). D2O (99.8%) was sourced from Cambridge Isotope Labs Inc, (Tewksbury, MA, USA). (NH4)2CO3 (99%), pepsin, gastric lipase (20,000 U/mg), cysteine (97%), bile salts, ethylbutyric acid (98%), N-methyl-N-t-butyldimethylsilyltrifluoroacetamide (95%), and imidazole (99%) were sourced from Sigma-Aldrich, (St. Louis, MO, USA).
7
Redox Signaling Compound Synthesis and Detection
8
Analytical Procedures for Carotenoids and Metabolites
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Astaxanthin, lutein, zeaxanthin, -cryptoxanthin, violaxanthin, echinenone (internal standard), -carotene and -carotene were provided by Hoffmann-La Roche (Basel, Switzerland). The fatty acid methyl esters (FAME) were purchased from Supelco (Bellefonte, PA, USA). Fructose, glucose, sucrose, acetonitrile, methanol, formic acid, chloroform, ethanol, tetrahydrofuran (THF), butylated hydroxytoluene, hexane, norvaline (IS), glutamine, cysteine, asparagine and tryptophan were procured from Sigma-Aldrich (St. Louis, MO, USA). ACCQ*TAG Ultra Derivatization kit for the AA determination was purchased from Waters (Milford, USA). The kit contained AA standard (Alanine, arginine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, valine, aspartic acid and glutamic acid), derivatising reagent (6-aminoquinolyl-Nhydroxysuccinimidyl carbamate), eluents and a sub-2 µm column for AA analysis. Water was supplied through a Milli-Q apparatus (Millipore, Milford, MA, USA).
9
Biochemical Analysis of Antioxidant Compounds
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Trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), gallic acid, acetic acid, propionic acid, butyric acid, Folin-Ciocalteu reagent, sodium hydroxide, hydrochloric acid, ferric chloride hexahydrate, sodium acetate, potassium chloride, potassium dihydrogen phosphate, sodium monohydrogen carbonate, sodium chloride, magnesium chloride hexahydrate, ammonium carbonate, calcium chloride dihydrate, sodium dihydrogen phosphate, tryptone, cysteine, sodium sulphide, resazurin, salivary α-amylase, porcine pepsin, bile acids ( porcine bile extract), pancreatin and ethanol were from Sigma-Aldrich (Darmstadt, Germany).
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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