A method of preparation and purification of (4R)-deuterated-reduced nicotinamide adenine dinucleotide phosphate.
Anal Biochem
; 221(2): 273-7, 1994 Sep.
Article
en En
| MEDLINE
| ID: mdl-7810866
(4R)-Deuterated-reduced nicotinamide adenine dinucleotide phosphate, (4R)-[2H]NADPH, was prepared by reduction of NADP+ using an NADP(+)-dependent alcohol dehydrogenase (EC 1.1.1.2) from Thermoanaerobium brockii and isopropanol-d8 as substrate at 43 degrees C, pH 9. More than 80% of the product was identified as reduced cofactor by reverse-phase (ODS) HPLC, and a 1H NMR study showed that all of the reduced cofactor was (4R)-deuterated. Less than 10% of the product was oxidized cofactor, the remainder being impurities from the breakdown of the dinucleotide compound. Subsequent purification carried out by semipreparative reverse-phase HPLC with 0.1 M NaCl at pH 8.5 gave a compound of more than 96% purity. Separated (4R)-[2H]NADPH fractions were freeze-dried and the white solid was stored at 5 degrees C with desiccant.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Deuterio
/
NADP
Idioma:
En
Revista:
Anal Biochem
Año:
1994
Tipo del documento:
Article
País de afiliación:
Australia
Pais de publicación:
Estados Unidos