Abstract
Publisher Summary This chapter discusses the synthesis of diphosphopyridine nucleotide from nicotinic acid. Synthesis of DPN from nicotinic acid in yeast, mammalian liver, and human erythrocytes has been shown to proceed by the three consecutive reactions. The intermediates, nicotinic acid mononucleotide (deamido-NMN) and nicotinic acid adenine dinucleotide (deamido-DPN), have been isolated from human erythrocytes after incubation with nicotinic acid. Nicotinic acid mononucleotide pyrophosphorylase activity is measured by following the formation of nicotinic acid- C 14 mono-nucleotide from nicotinic acid-7- C 14 with a strip counter after separation of reaction components by paper chromatography. Maximal activity of the enzyme has been observed only in the presence of magnesium, orthophosphate, and ATP. Deamido-DPN and DPN pyrophosphorylase activities are closely associated throughout fractionation of an extract of acetone-dried hog liver. The most purified preparation of deamido-DPN pyrophosphorylase has been obtained for purifying DPN pyrophosphorylase from hog liver extracts. Deamido-DPN pyrophosphorylase activity has also been demonstrated in extracts of human erythrocyte and in yeast autolyzates.
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