Methanol-d4 (MeOD, 99.8% atom D) and sodium deuteroxide (NaOD, 99.5% atom D; 40% in D2O) were obtained from Cambridge Isotope Laboratories, Inc. (Andover, MA, USA). Deuterium oxide (D2O, 99.9% atom D) including 0.05% 3-(trimethylsilyl) propionic-2, 2, 3, 3-d4 acid sodium salt (TSP) and potassium phosphate (KH2PO4) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
Sodium deuteroxide
Manufactured by Cambridge Isotopes
Sourced in United States
Sodium deuteroxide is a stable isotope of sodium, with the chemical formula NaOD. It is commonly used as a source of deuterium, which is a heavy isotope of hydrogen. Sodium deuteroxide is a white crystalline solid that is soluble in water and other polar solvents.
Automatically generated - may contain errors
Lab products found in correlation
Deuterium chloride, by Cambridge Isotopes (4 mentions)
Deuterium oxide, by Cambridge Isotopes (4 mentions)
3 trimethylsilyl propionic 2 2 3 3 d4 acid sodium salt tsp, by Merck Group (1 mentions)
Potassium phosphate kh2po4, by Merck Group (1 mentions)
Methanol d4, by Cambridge Isotopes (1 mentions)
Deuterium oxide, by Merck Group (1 mentions)
Chelex resin, by Bio-Rad (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Deuterium oxide, by Fujifilm (1 mentions)
Methanol d4, by Kanto Chemical (1 mentions)
Bovine serum albumin (bsa), by Promega (1 mentions)
Chloramphenicol, by Merck Group (1 mentions)
Ampicillin, by ICN Biomedicals (1 mentions)
Pad2hgdh pet20b plasmid, by GenScript (1 mentions)
D malate, by Thermo Fisher Scientific (1 mentions)
Luria bertani agar, by Merck Group (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
7 protocols using sodium deuteroxide
1
NMR Spectroscopy Reagent Preparation
2
Preparation and Characterization of Compounds
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Compounds KMG-301,7 (link)2 ,15 and 2a (15 ) were prepared according to reported
procedures, and their purity (>98%) was verified by reverse phase
C-18 HPLC. High-purity piperazine-N,N′-bis(2-ethanesulfonic acid) (PIPES), 37% HCl, 45% KOH, and
99.999% anhydrous MgCl2 were purchased from Aldrich. High
purity 99.999% KCl was obtained from Alfa Aesar. ATP was obtained
from Aldrich as the disodium salt. Deuterium oxide (D, 99.9%), sodium
deuteroxide (D, 99.5%, 40% w/w in D2O), and deuterium chloride
(D, 99.5%, 35% w/w in D2O) were obtained from Cambridge
Isotope Laboratories. All aqueous solutions for fluorescence experiments
were prepared using deionized water having a resistivity of ≥18
MΩ/cm. Other solvents were obtained from commercial suppliers
and used as received. Aqueous buffers were treated with Chelex resin
(Bio-Rad) according to the manufacturer’s protocol to remove
adventitious metal ions.
procedures, and their purity (>98%) was verified by reverse phase
C-18 HPLC. High-purity piperazine-N,N′-bis(2-ethanesulfonic acid) (PIPES), 37% HCl, 45% KOH, and
99.999% anhydrous MgCl2 were purchased from Aldrich. High
purity 99.999% KCl was obtained from Alfa Aesar. ATP was obtained
from Aldrich as the disodium salt. Deuterium oxide (D, 99.9%), sodium
deuteroxide (D, 99.5%, 40% w/w in D2O), and deuterium chloride
(D, 99.5%, 35% w/w in D2O) were obtained from Cambridge
Isotope Laboratories. All aqueous solutions for fluorescence experiments
were prepared using deionized water having a resistivity of ≥18
MΩ/cm. Other solvents were obtained from commercial suppliers
and used as received. Aqueous buffers were treated with Chelex resin
(Bio-Rad) according to the manufacturer’s protocol to remove
adventitious metal ions.
3
Metabolite Identification Protocol
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The solvents—methanol, chloroform, and chloroform-d (CDCl3) were purchased from Millipore Sigma (St. Louis, MO, USA). Deuterated chemicals (deuterium oxide, D2O; deuterium chloride, DCl; and sodium deuteroxide, NaOD) were commercially available and purchased from Cambridge isotope laboratories (Tewksbury, MA, USA). Internal standard for metabolite quantification, 3-(trimethylsilyl)-1-propane sulfonic acid-d6 sodium salt (DSS-d6) was purchased from Chenomx Inc. (Edmonton, AB, Canada).
4
Kinetic Study of Hydrolysis Reactions
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Kinetic data were collected by recording the 1H-NMR (400 MHz) spectra at 2- min intervals. Data were obtained at 70.0 °C under a N2 flow of 400 L/h. Deuterated solvents were used for all measurements: Deuterium oxide, 99.9% D from Wako; methanol-d4, 99.8% D from Kanto Chemical Co. Inc.; 1,4-dioxane-d8, 99% D from Cambridge Isotope Laboratories, Inc.; and sodium deuteroxide, 40% wt. % in D2O, 99.5% D from Sigma-Aldrich, Co. Parameters were set using the reaction solution at room temperature. Reactions were initiated by raising the temperature of the NMR machine. Spectral data were then recorded for at least one half-life. Hydrolysis experiments were repeated three times. Hydrolysis products were identified by comparison of the 1H-NMR data with those of authentic samples, or by MS-ESI. Pseudo-first-order rate constants (kobs) were evaluated by a linear least-squares fitting of a plot of the logarithm of reactant concentration versus time.
5
Recombinant PA0317 Enzyme Production
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The PaD2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript. The plasmid was sequenced to verify the presence of the wildtype gene. Escherichia coli strain Rosetta(DE3)pLysS was from Novagen. Bovine serum albumin was purchased from Promega. Luria–Bertani agar, Luria–Bertani (LB) broth, chloramphenicol, IPTG, lysozyme, PMS, and PMSF were obtained from Sigma–Aldrich. Ampicillin was purchased from ICN Biomedicals. d -2-hydroxyglutarate was purchased from MilliporeSigma. D-malate was purchased from Alfa Aesar. Deuterium oxide, deuterium chloride, and sodium deuteroxide were purchased from Cambridge Isotope Laboratories, Inc. Glucose, glycerol, and all the other reagents were of the highest purity commercially available.
6
NMR Solvent Characterization Protocol
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The deuterated solvents used for 1H NMR tests, including deuterium oxide (D2O, CAS No. 7789-20-0, >99.5%), deuterium chloride (DCl, CAS No. 7698-05-7, >99.9%), and sodium deuteroxide (NaOD, CAS No. 14014-06-3, >99.5%) were purchased from Cambridge Isotope Laboratories Inc. (Andover, MA, USA). The reference standard L-cystine (CAS No. 56-89-3, ≥98%) and the internal standard (ISTD) maleic acid (CAS No. 110-16-7, ≥99%) were obtained from Sigma-Aldrich (Saint Louis, MO, USA). ISTD was diluted to 60 µL/mL using either D2O, DCl, or NaOD before use. Unless otherwise specified, other reagents and solvents were of analytical grade, which were purchased from Shandong Yuwang Industrial Co., Ltd. (Yucheng, China).
7
Peptide and Lipid Preparation for NMR Analysis
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