Regulation in tobacco callus of enzyme activities of the nicotine pathway

In tobacco callus, the induction of nicotine synthesis, which stimulates enzyme activities of the ornithine-methylpyrroline route (see the preceding paper), also leads to marked changes in the enzyme activities of the pyridine-nucleotide cycle. This cycle provides the metabolite (probably nicotinic acid) for condensation with methylpyrroline to produce nicotine. The activities of eight enzymes of the pyridine-nucleotide cycle and of quinolinic-acid phosphoribosyltransferase, the anaplerotic enzyme, were determined by high-performance liquid chromatography assays. The distinct changes of their activities upon induction of nicotine synthesis lead to the following conclusions: i) nicotinic acid is the relevant metabolite which is provided by the pyridine-nucleotide cycle and consumed for nicotine synthesis. ii) The enhancement of the nicotinic-acid pool arises in two ways, by synthesis of NAD and degradation via nicotinamide mononucleotide and by a direct route from nicotinic-acid mononucleotide (NaMN) which is degraded by a glycohydrolase with a rather high K _m value. Such a K _m value prevents the complete depletion of the NaMN pool.Abbreviations HPLC high-performance liquid chromatography - NAD-PPase NAD-pyrophosphatase - NaMN-ATase nicotinic-acid mononucleotide (NaMN) adenylyltransferase - NaMN-GHase NaMN-glycohydrolase - Na-PRTase nicotinic-acid phosphoribosyltransferase - NMN-ATase nicotinamide mononucleotide (NMN) adenylyltransferase - NMN-Ghase NMN-glycohydrolase - PMT putrescine methyltransferase - Qa-PRTase quinolinic acid phosphoribosyltransferase