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A high yield microscale enzymatic synthesis and purification Of 14 C‐Labeled Nicotinamide Adenine Dinucleotide Phosphate (NADP + )
Author(s) -
Ronneberg Andre,
Metz Gordon,
Weld Richard,
Roffey Peter,
Craney Chris
Publication year - 1992
Publication title -
journal of labelled compounds and radiopharmaceuticals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 47
eISSN - 1099-1344
pISSN - 0362-4803
DOI - 10.1002/jlcr.2580310408
Subject(s) - chemistry , nicotinamide adenine dinucleotide , nad+ kinase , nicotinamide adenine dinucleotide phosphate , phosphate , chromatography , elution , enzyme , nicotinamide , biochemistry , oxidase test
Uniformly labeled (U) 14 C nicotinamide adenine dinucleotide phosphate (NADP + ) was synthesized by phosphorylating [U‐ 14 C]nicotinamide adenine dinucleotide (NAD + ) in the presence of immobilized NAD + kinase. The 15 μCi (600 μL) synthesis consistently achieved yields between 80% and radiochemical purities greater than 95%. The [U‐ 14 C]NADP + was purified by high performance anion‐exchange chromatography using a gradient elution of ammonium bicarbonate. This procedure may be applicable to the synthesis of other charged, UV‐absorbing products of enzyme‐catalyzed reactions.