4-Phenylcatechol (2 mM) was combined with excess AphC in 20 mM Hepes (I = 0.1 M), pH = 8. The reaction was incubated at room temperature for 30 min, and the enzyme was removed by filtration through a 30 kDa molecular weight cut-off membrane. The filtrate was supplemented with 200 mM of NH4OH and incubated overnight at room temperature. The resultant reaction mixture was freeze dried and suspended in 0.5 ml of D2O for NMR analysis. NMR spectra were recorded using a Bruker AVANCE III 850 spectrometer. Data were analyzed using ACD NMR Processor Academic Edition.
Avance 3 850 spectrometer
Manufactured by Bruker
Sourced in Germany
The AVANCE III 850 spectrometer is a high-performance nuclear magnetic resonance (NMR) instrument designed for advanced research and analysis. It features a superconducting magnet capable of generating a magnetic field strength of 20 Tesla, enabling the acquisition of high-resolution NMR data. The spectrometer is capable of operating at a proton frequency of 850 MHz, making it suitable for a wide range of applications in chemistry, materials science, and life sciences.
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Lab products found in correlation
2 protocols using avance 3 850 spectrometer
1
Enzymatic Conversion of 4-Phenylcatechol
2
Intramolecular Deuterium Abundance Analysis
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For NMR measurements of intramolecular deuterium abundances, each sample of the glucose derivative prepared as described above was dissolved in a mixture of 83% v/v acetonitrile, 17% C6F6, and 0.01% C6D6, then transferred to a 5-mm NMR tube with a PTFE valve (J. Young Scientific Glassware Ltd., Windsor, U.K.) containing ca. 5 mg of NaHCO3. Deuterium NMR spectra were acquired and processed as previously described64 (link), using an AVANCE III 850 spectrometer (Bruker BioSpin GmbH, Rheinstetten, Germany) equipped with a 19F lock and a cryogenic probe optimized for deuterium detection. Deuterium NMR spectra were integrated by deconvolution with a Lorentzian line shape fit, using TopSpin™ 3.2 (Bruker BioSpin GmbH, Rheinstetten, Germany). The D6S/D6R isotopomer ratio was determined as the ratio of the integrals of the D6S and D6R signals26 (link). For each sample, five to eight spectra were recorded and the average D6S/D6R ratio was calculated.
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