L methionine

Manufactured by Formedium

Sourced in United Kingdom

L-methionine is an essential amino acid that serves as a building block for proteins. It plays a crucial role in various metabolic processes within the body.

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2 protocols using l methionine

Gluconeogenic Precursor Screening in Hepatocytes

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Twenty-four hours after isolation, primary hepatocytes (plated in 12-well plates; 2.5 × 10⁵ cells per well) were incubated for 9 hours with Dulbecco modified Eagle medium lacking glucose, pyruvate, glutamine, and phenol red (Gibco, Waltham, MA; A14430-01). Then cells were washed with PBS and treated for 20 hours with medium containing combinations of glucagon (100 nmol/L) and corticosterone (1 μmol/L) together with one gluconeogenic precursor (20 mmol/L): pyruvate (Biological Industries; cat# 03-042-1B), glycerol (Biolab; cat# 07120501), L-ornithine (Sigma-Aldrich; cat# O2375-5G), L-asparagine (Formedium, Norfolk, UK; DOC0114), L-tyrosine (Formedium; DOC0190), L-methionine (Formedium; DOC0166), L-aspartic-acid (Formedium; DOC0166), L-serine (Formedium; DOC0178), L-alanine (Formedium; DOC0102), L-histidine (Formedium; DOC0142), L-phenylalanine (Formedium; DOC0170). After 20 hours, 40 μL of medium was sampled, and glucose was measured using the glucose oxidase colorimetric method according to the manufacturer’s instructions (Sigma-Aldrich; GAGO20).

Korenfeld N., Finkel M., Buchshtab N., Bar-Shimon M., Charni-Natan M, & Goldstein I. (2021). Fasting Hormones Synergistically Induce Amino Acid Catabolism Genes to Promote Gluconeogenesis. Cellular and Molecular Gastroenterology and Hepatology, 12(3), 1021-1036.

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Cultivation of Bacteroides thetaiotaomicron

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B. thetaiotaomicron was routinely grown on brain- heart infusion (BHI, OXOID) supplemented with hemin (1 μg/ml, Sigma-Aldrich). Agar (Fisher bioreagents), erythromycin (25 μg/ml, Duchefa Biochemies), gentamicin (200 μg/ml, Formedium) and 5-fluoro-2′-deoxyuridine (FUdR, 200 μg/ml, Sigma-Aldrich) were added when required. Minimal medium (MM) contained 7.5 mM NH₄SO₄ (Thermo Scientific), 9.5 mM Na₂CO₃ (Melford), 4 mM L-Cysteine (Melford), 100 mM KH2PO4 (pH 7.2)(Fisher Chemical), 1 μg/ml menadione (Sigma-Aldrich), 1 μg/ml hemin and a mixture of mineral salts (15 mM NaCl (Duchefa Biochemies), 0.18 mM CaCl₂•2H₂0 (Fisher Chemical), 0.1 mM MgCl2•6H₂O (Fisher Chemical), 0.5 mM MnCl₂•4H₂O (Acros Organics) and 0.04 mM CoCl2•6H₂O (Sigma-Aldrich)). Vitamin B₁₂ (Sigma-Aldrich) and L-methionine (Formedium) were added as required and described in the text. 0.5% of fructose (Thermo scientific) was added as carbon source. Cells were grown in a Whitley A35 anaerobic workstation with a mixture of gas of 80% N₂, 10% CO₂ and 10% H₂.

Abellon-Ruiz J., Jana K., Silale A., Frey A.M., Baslé A., Trost M., Kleinekathöfer U, & van den Berg B. (2023). BtuB TonB-dependent transporters and BtuG surface lipoproteins form stable complexes for vitamin B12 uptake in gut Bacteroides. Nature Communications, 14, 4714.

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