L leucine

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

L-leucine is a naturally occurring amino acid that plays a key role in protein synthesis and cellular function. It is an essential nutrient required for various physiological processes. As a laboratory reagent, L-leucine is commonly used in cell culture media and other biochemical applications.

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Lab products found in correlation

Maltodextrin, by Merck Group (2 mentions) Reagent grade ethanol, by Merck Group (2 mentions) Sucrose, by Thermo Fisher Scientific (2 mentions) Methanol, by Avantor (2 mentions) Hydrochloric acid (hcl), by Avantor (2 mentions) Ethanol, by Avantor (2 mentions) Potassium phosphate monobasic, by Thermo Fisher Scientific (2 mentions) L phenylalanine, by Thermo Fisher Scientific (2 mentions) Thiamine, by Thermo Fisher Scientific (1 mentions) Yeast extract, by Thermo Fisher Scientific (1 mentions) Tetracycline, by Merck Group (1 mentions) H3bo3, by Merck Group (1 mentions) Feso4 7h2o, by Merck Group (1 mentions) Mnso4 h2o, by Merck Group (1 mentions) K2hpo4 3h2o, by Merck Group (1 mentions) Na2hpo4 12h2o, by Merck Group (1 mentions) Mgso4 7h2o, by BD (1 mentions) Cacl2 2h2o, by Thermo Fisher Scientific (1 mentions) Znso4 7h2o, by Avantor (1 mentions) Nh4 2so4, by Merck Group (1 mentions)

16 protocols using l leucine

Optimized Minimal Media Formulation

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The composition of the aqueous phase in all experiments was either luria broth or as described by Wubbolts et al30 (link). KH₂PO₄, 4 g/L; K₂HPO₄ (3H₂O), 15.9 g/L; Na₂HPO₄ (12H₂O), 7 g/L; (NH₄)₂SO₄, 1.2 g/L; NH₄Cl, 0.2 g/L (all from Sigma Aldrich); yeast extract (Oxoid), 5 g/L; L-leucine, 0.6 g/L; L-proline, 0.6 g/L; thiamine, 5 mg/L (All from Alfa Aesar). After autoclaving, MgSO₄ (7H₂O) (BDH), 1 g/L (BDH); 1 mL of trace minerals (composition below); 1 mL of 4% (w/v) CaCl₂(2H₂O) (Alfa Aesar) and 10 g/L D-glucose (Sigma Aldrich) were added having all been heat sterilized separately. 1 ml of 10 mg/ml filter sterilised tetracycline (Sigma Aldrich) was also added.
The solution of trace minerals contained per liter of 5 M HCL: FeSO₄ (7H₂O) (Sigma Aldrich), 40 g; MnSO₄ (H₂O) (Sigma Aldrich), 10 g; CoCl₂ (6H₂O) (Fluka), 4.75 g; ZnSO₄ (7H₂O) (VWR), 2 g; MoO₄Na₂ (2H₂O) (Sigma Aldrich), 2 g; CuCl₂, (2H₂O) (Riedel-de Haen), 1 g; H₃BO₃ (Sigma Aldrich), 0.50 g.

Grant C., Deszcz D., Wei Y.C., Martínez-Torres R.J., Morris P., Folliard T., Sreenivasan R., Ward J., Dalby P., Woodley J.M, & Baganz F. (2014). Identification and use of an alkane transporter plug-in for applications in biocatalysis and whole-cell biosensing of alkanes. Scientific Reports, 4, 5844.

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Spectinamide 1599·2HCl Synthesis and Materials

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Spectinamide 1599·2HCl as a hydrate with 2.1–2.7 H₂O content was synthesized as previously described (19 (link)). L-leucine was purchased from Alfa Aesar (Ward Hill, MA, USA). Dosator supplies were purchased from McMaster-Carr (Atlanta, GA, USA)

Stewart I.E., Lukka P.B., Liu J., Meibohm B., Gonzalez-Juarrero M., Braunstein M.S., Lee R.E, & Hickey A.J. (2019). Development and Characterization of a Dry Powder Formulation for Anti-Tuberculosis Drug Spectinamide 1599. Pharmaceutical research, 36(9), 136.

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Bedaquiline Fumarate Formulation Protocol

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Bedaquiline fumarate (BDQ) was purchased from BOC Sciences (Shirley, NY, USA). 2-hydroxypropyl-β-cyclodextrin was purchased from Cayman Chemical Company (Ann Arbor, MI, USA). L-leucine (99%) was purchased from Alfa Aesar (Ward Hill, MA, USA). Sucrose, common lab salts, and common solvents were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Heparin for maintaining intravenous catheter patency was purchased from APP Pharmaceuticals (Schaumburg, IL, USA). Unless otherwise specified, all reagents were used as received without further purification. Nitrogen (N2) gas cylinders were purchased from Airgas National Welders (Raleigh, NC, USA).

Maloney S.E., Stewart I.E., Podell B.K., Gary H.E., Mecham J.B., Berube B.J., Baldwin S.L., Coler R.N, & Hickey A.J. (2023). Preparation Strategies of the Anti-Mycobacterial Drug Bedaquiline for Intrapulmonary Routes of Administration. Pharmaceuticals, 16(5), 729.

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Synthesis and Characterization of n-Propyl Pyrazinoate

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n-Propyl pyrazinoate (PAE) used in this study was prepared by previously established methods and confirmed by NMR [51 (link)]. Maltodextrin, POA and reagent grade ethanol were obtained from Sigma-Aldrich (St. Louis, MO). L-leucine was obtained from Alfa Aesar (Ward Hill, MA). PZA was obtained from ACROS Organics(Morris Plains, NJ).

Montgomery S.A., Young E.F., Durham P.G., Zulauf K.E., Rank L., Miller B.K., Hayden J.D., Lin F.C., Welch J.T., Hickey A.J, & Braunstein M. (2018). Efficacy of pyrazinoic acid dry powder aerosols in resolving necrotic and non-necrotic granulomas in a guinea pig model of tuberculosis. PLoS ONE, 13(9), e0204495.

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Intracellular M. tuberculosis infection

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The M. tuberculosis H37Rv-derived auxotroph strain mc²6206 (Sampson et al. 2004 (link), Mouton et al. 2019 (link)) was grown in Middlebrook 7H9 medium (BD Biosciences, Franklin Lakes, NJ) supplemented with 0.2% glycerol (Fisher Chemical, Waltham, MA), 0.05% Tween-80 (Acros Organics, Fair Lawn, NJ), 10% OADC (BD Biosciences), 24 μg/ml d-pantothenic acid (Alfa Aesar), and 50 μg/ml l-leucine (Alfa Aesar). Liquid Mtb cultures were maintained at 37°C with slow shaking (50 rpm).
Mtb mc²6206 growing in log-phase was quantified by optical density measurement at 600 nm using the conversion of OD 1 = 3 × 10⁸ Mtb bacteria per ml. The number of bacteria required for an MOI of 10 was washed and resuspended in RPMI 1640 cell culture media without antibiotics. Bacteria were added to the differentiated THP–WT, THP–△PPM1A, or THP–PPM1A⁺ macrophages and incubated at 37°C for 16 h. Three phosphate buffered saline (PBS) washes were then performed to remove extracellular, nonphagocytosed bacteria and infection was continued at 37°C for a total infection time of 48 h. Noninfected control cells underwent identical wash and incubation steps.

Madden K., El Hamra R., Berton S., Felker J., Alvarez G.G., Blais A, & Sun J. (2023). Mycobacterium tuberculosis infection triggers epigenetic changes that are enriched in a type I IFN signature. microLife, 4, uqad006.

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Synthesis of n-Propyl Pyrazinoate (PAE)

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n-Propyl pyrazinoate (PAE) was synthesized for these experiments using a method previously employed by one of the authors (Cynamon et al., 1992 (link)) and confirmed by proton NMR. Maltodextrin, POA and reagent grade ethanol were obtained from Sigma-Aldrich (St. Louis, MO, USA). L-leucine was obtained from Alfa Aesar (Ward Hill, MA, USA).

Durham P.G., Young E.F., Braunstein M.S., Welch J.T, & Hickey A.J. (2016). A dry powder combination of pyrazinoic acid and its n-propyl ester for aerosol administration to animals. International journal of pharmaceutics, 514(2), 384-391.

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Synthesis and Characterization of Antibacterial Compounds

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Tigecycline (TGC) was purchased from ChemShuttle (Burlingame, CA).
Lactose monohydrate (LAC) was purchased from Mallinckrodt, Inc. (Paris, KY).
l-leucine (99%) (LEU) was purchased from Alfa Aesar (Ward Hill,
MA). Common laboratory salts and solvents were acquired from Thermo Fisher
Scientific (Waltham, MA). Unless otherwise specified, all reagents were used as
received without further purification. Nitrogen (N₂) gas cylinders
were purchased from Airgas National Welders (Raleigh, NC).

Rowan N.J., MacGregor S.J., Anderson J.G., Fouracre R.A., McIlvaney L, & Farish O. (1999). Pulsed-Light Inactivation of Food-Related Microorganisms. Applied and Environmental Microbiology, 65(3), 1312-1315.

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Nanoparticle-Mediated Gene Delivery Protocol

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Triphosgene was bought from TCI (Tokyo, Japan), hexamethyldisilazane (HMDS), H-Lys(Z)-OH (N6-carbobenzyloxy-l-lysine), and l-leucine were purchased from Alfa Aesar (Haverhill, MA, USA). Tetrahydrofuran (THF), dimethylformamide (DMF), and diethyl ether were bought from Sinopharm Chemical Reagent Co. (Shanghai, China), Ltd. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Hoechst 33342, hydrogen bromide (33% in acetic acid), chloroquine phosphate, trypsin-EDTA solution 0.25%, branched polyethylenimine (PEI, M_n = 10 kDa, M_w = 25 kDa, impurities ≤1% water), and rhodamine B isothiocyanate were bought from Sigma-Aldrich (St. Louis, MO, USA). BCA protein kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Luciferase assay kit and plasmid DNA (pDNA) pGL3-control vector (5256 bp) for transfection were bought from Promega (Madison, WI, USA). All the cells were obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China).

Zhang Y., Zhou Z, & Chen M. (2018). The Length of Hydrophobic Chain in Amphiphilic Polypeptides Regulates the Efficiency of Gene Delivery. Polymers, 10(4), 379.

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Dry Powder Inhaler Formulation Development

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Mannitol was supplied from Roquette (Lestrem, France; purity 99.9%), salbutamol sulfate from L.B. (Bohle, Germany; purity, 99.9%), lactose from DFE Pharma (Gosh, Germany; purity, 99.9%), and L-leucine (purity, 99.9%) and the monobasic potassium phosphate by Acros Organics (Geel, Belgium; purity, 99.9%). All solvents (m ethanol, ethanol, and hydrochloric acid) were purchased from VWR International Ltd. (Leighton Buzzard, UK) and were high-performance liquid chromatography (HPLC) grade. Fisher Scientific™ (UK) provided the 0.22-μm filters.

Ferdynand M.S, & Nokhodchi A. (2020). Co-spraying of carriers (mannitol-lactose) as a method to improve aerosolization performance of salbutamol sulfate dry powder inhaler. Drug Delivery and Translational Research, 10(5), 1418-1427.

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Dry Powder Inhaler Formulation Composition

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α-Lactose monohydrate (DFE Pharma, Gosh, Germany, purity 99 + %; lactose is one of the main excipients approved to be used in DPI formulation), salbutamol sulfate (L.B. Bohle, Germany; purity 99%), and _L-leucine (Acros Organics, Geel, Belgium, purity 99 + %; it is used as anti-adherent and lubricant) were used in their purest form. Monobasic potassium phosphate (Fisher Scientific, UK) was used for the preparation of the mobile phase for high-pressure liquid chromatography (HPLC). All solvents (m ethanol, ethanol, and hydrochloric acid) were purchased from VWR International Ltd. (Leighton Buzzard, UK) and were of HPLC grade.

Molina C., Kaialy W., Chen Q., Commandeur D, & Nokhodchi A. (2017). Agglomerated novel spray-dried lactose-leucine tailored as a carrier to enhance the aerosolization performance of salbutamol sulfate from DPI formulations. Drug Delivery and Translational Research, 8(6), 1769-1780.

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