Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The pyridine-nucleotide cycle in tobacco

Enzyme activities for the recycling of NAD


In order to elucidate the NAD-recycling pathway the following enzyme activities have been characterized in different tobacco tissues and in tomato root: NAD pyrophosphatase, nicotinamide mononucleotide (NMN)/nicotinic acid mononucleotide (NaMN) glycohydrolases, nicotinamidase and nicotinic acid phosphoribosyltransferase. The investigations were performed with protein extracts purified by gel filtration and enzymatic activities were determined by high-performance liquid chromatography methods. The kinetic parameters of the different enzymes from tobacco root and their specificity are reported. The data are in favor of the so-called pyridine-nucleotide cycle VI (NAD→NMN→nicotinamide→nicotinic acid→NaMN→nicotinic acid adenine dinucleotide→NAD). In the nicotine-producing tobacco root a further direct route leading from NaMN to nicotinic acid is proposed. These data are reconciled with the assumption that it is nicotinic acid which is provided by the pyridine-nucleotide cycle for the synthesis of nicotine.

This is a preview of subscription content, log in to check access.



high-performance liquid chromatography


nicotinic acid


nicotinic acid adenine dinucleotide


nicotinic acid mononucleotide


nicotinamide mononucleotide




  1. Andreoli, A.J., Okita, T.W., Bloom, R., Grover, T.A. (1972) The pyridine nucleotide cycle: presence of nicotinamide mononucleotide-specific glycohydrolase in Escherichia coli. Biochem. Biophys. Res. Commun. 49, 264–269

  2. Ashton, A.R., Polya, G.M. (1975) Higher plant cyclic nucleotide phosphodiesterases: Resolution, partial purification and properties of three phosphodiesterases from potato tuber. Biochem. J. 149, 329–339

  3. Bartkiewicz, M., Sierakowska, H., Shugar, D. (1984) Nucleotide pyrophosphatase from potato tubers. Purification and properties. Eur. J. Biochem. 143, 419–426

  4. Barz, W. (1985) Metabolism and degradation of nicotinic acid in plant cell cultures. In: Primary and secondary metabolism of plant cell cultures, pp. 186–195, Neumann, K.H., Barz, W., Reinhard, E., eds. Springer, Berlin Heidelberg New York

  5. Bischoff, E., Tran-Thi, T.-A., Decker, K.F.A. (1975) Nucleotide pyrophosphatase of rat liver: A comparative study on the enzymes solubilized and purified from plasma membrane and endoplasmic reticulum. Eur. J. Biochem. 51, 353–361

  6. Bush, L.P., Saunders, J.L. (1977) Accumulation, manipulation and regulation of nicotine content in tobacco. Tob. Chem. Symp. (Am. Chem. Soc.) New Orleans, pp. 389–415

  7. Clayton, R.A., Hanselman, L.M. (1960) Tobacco nucleotide pyrophosphatases. Arch. Biochem. Biophys. 87, 161–166

  8. Elliot, G.C. (1982) Evidence for a physiological active nicotinamide phosphoribosyltransferase in cultured human fibroblasts. Biochem. Biophys. Res. Commun. 104, 996–1002

  9. Foster, J.W., Moat, A.G. (1980) Nicotinamide adenine dinucleotide biosynthesis and pyridine nucleotide cycle metabolism in microbial systems. Microbiol. Rev. 44, 83–105

  10. Gholson, R.K. (1966) The pyridine nucleotide cycle. Nature 212, 933–935

  11. Godavari, H.R., Waygood, E.R. (1970) Nicotinamide adenine dinucleotide metabolism in plants. I. Intermediates of biosynthesis in wheat leaves and the effect of benzimidazole. Can. J. Bot. 48, 2267–2278

  12. Gopinathan, K.P., Sirsi, M., Vaidyanathan, C.S. (1964) Nicotinamide-adenine dinucleotide glycohydrolase of Mycobacterium tuberculosis H37Rv. Biochem. J. 91, 277–282

  13. Hanna, L.S., Hess, S.L., Sloan, D.L. (1983) Kinetic analysis of nicotinate phosphoribosyltransferase from yeast using high pressure liquid chromatography. J. Biol. Chem. 258, 9745–9754

  14. Hanna, L., Sloan, D.L. (1980) A high-pressure liquid chromatography procedure for monitoring nicotinate phosphoribosyltransferase activity. Anal. Biochem. 103, 230–234

  15. Hayakawa, T., Shibata, K., Iwai, K. (1984) Nicotinate phosphoribosyltransferase from hog liver: regulatory effect of ATP at a physiological concentration of 5-phosphoribosyl-1-pyrophosphate. Agric. Biol. Chem. 48, 455–460

  16. Hillyard, D., Rechsteiner, M., Manlapaz-Ramos, P., Imperial, J.S., Cruz, L.J., Olivera, B.M. (1981) The pyridine nucleotide cycle; studies in Escherichia coli and the human cell line D98/AH2. J. Biol. Chem. 256, 8491–8497

  17. Hilz, H., Koch, R., Fanick, W., Klapproth, K., Adamietz, P. (1984) Nonenzymatic ADP-ribosylation of specific mitochondrial polypeptides. Proc. Natl. Acad. Sci. USA 81, 3929–3933

  18. Imai, T. (1979) Isolation and properties of a glycohydrolase specific for nicotinamide mononucleotide from Azotobacter vinelandii. J. Biochem. 85, 887–899

  19. Imsande, J. (1961) Pathway of diphosphopyridine nucleotide biosynthesis in Escherichia coli. J. Biol. Chem. 236, 1494–1497

  20. Kole, E., Sierakowska, H., Shugar, D. (1976) Novel activity of potato nucleotide pyrophosphatase. Biochim. Biophys. Acta 438, 540–550

  21. Kornberg, A., Pricer, Jr., W.E. (1950) Nucleotide pyrophosphatase. J. Biol. Chem. 182, 763–778

  22. Mann, D.F., Byerrum, R.U. (1974) Activaton of the de novo pathway for pyridine nucleotide biosynthesis prior to ricine biosynthesis in Castor beans. Plant Physiol. 53, 603–609

  23. Marushige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497

  24. Meyer, R. (1984) Die Ermittlung des Nucleotid pools in der pflanzlichen Zelle: Methodik und Anwendung. Thesis, Technische Universität, Braunschweig

  25. Mizusaki, S., Tanabe, Y., Kisaki, T., Tamaki, E. (1970) Metabolism of nicotinic acid in tobacco plants. Phytochemistry 9, 549–554

  26. Ohtsu, E., Nishizuka, Y. (1971) Nicotinamide phosphoribosyltransferase and NAD pyrophosphorylase from Lactobacillus fructosus. Methods Enzymol. 18 B, 127–132

  27. Olivera, B.M., Ferro, A.M. (1982) Pyridine nucleotide metabolism and ADP-ribosylation. In: ADP-ribosylation reactions; biology and medicine, pp. 19–40, Hayaishi, O., Ueda, K., eds. Academic Press, New York London

  28. Petrack, B., Greengard, P., Craston, A., Sheppy, F. (1965) Nicotinamide deamidase from mammalian liver. J. Biol. Chem. 240, 1725–1730

  29. Pietta, P., Pace, M., Menegus, F. (1983) High performance liquid chromatography for assaying NAD glycohydrolase from Neurospora crassa conidia. Anal. Biochem. 131, 533–537

  30. Razzel, W.E. (1966) Plant tissue phosphodiesterases. Biochem. Biophys. Res. Commun. 22, 243–247

  31. Roberts, D.W.A. (1959) The hydrolysis of diphosphopyridine nucleotide by juice expressed from wheat leaves. J. Biol. Chem. 234, 655–657

  32. Ryrie, I.J., Scott, K.J. (1969) Nicotinate, quinolinate and nicotinamide as precursors in the biosynthesis of nicotinamideadenine dinucleotide in barley. Biochem. J. 115, 679–685

  33. Sarma, D.S.R., Rajalakshmi, S., Sarma, P.S. (1961) Deamidation of nicotinamide and NMN. Biochem. Biophys. Res. Commun. 6, 389–393

  34. Sarma, D.S.R., Rajalakshmi, S., Sarma, P.S. (1964) Studies on the enzymes involved in nicotinamide adenine dinucleotide metabolism in Aspergillus niger. Biochim. Biophys. Acta 81, 311–322

  35. Sloan, D.L., Ali, L.Z., Aybar-Batista, D., Yan, C., Hess, S.L. (1984) Enzymatic assay procedures that employ high-performance liquid chromatography: competition between phosphoribosyltransferases for a common substrate. J. Chromatiogr. 316, 43–52

  36. Su, S., Chaykin, S. (1971) Nicotinamide deamidase. Methods Enzymol. 18B, 185–192

  37. Thomas, E., Davey, M.R. (1975) From single cells to plants, Wykeham Publications, London

  38. Thomzik, J. (1983) Untersuchungen zum Stoffwechsel von Nicotinsäure und ihren Derivaten in pflanzlichen Zellsuspensionskulturen und Pflanzen. Thesis, Universität Münster

  39. Wagner, R. (1985) Der Pyridin-Nucleotid-Cyclus und die Regulation der Nicotinbiosynthese in Tabak. Thesis, Technische Universität Braunschweig

  40. Wagner, R., Wagner, K.G. (1984) Determination of quinolinic acid phosphoribosyltransferase in tobacco. Phytochemistry 23, 1881–1883

  41. Wagner, R., Wagner, K.G. (1985) The pyridine nucleotide cycle in tobacco. Enzyme activities for the de novo synthesis of NAD. Planta 165, 532–537

  42. Waller, G.R., Dermer, O.C. (1981) Enzymology of alkaloid metabolism in plants and microorganisms. Biochem. Plants 7, 317–402

  43. Waller, G.R., Yang, K.S., Gholson, R.K., Hadwiger, L.A., Chaykin, S. (1966) The pyridine nucleotide cycle and its role in the biosynthesis of ricine by Ricinus communis L. J. Biol. Chem. 241, 4411–4418

  44. Willmitzer, L. (1979) Demonstration of in vitro covalent modification of chromosomal proteins by poly (ADP) ribosylation in plant nuclei. FEBS Lett. 108, 13–16

  45. Willmitzer, L., Wagner, K.G. (1982) Poly (ADP-ribose) synthesis in plants. In: ADP-ribosylation reactions, biology and medicine, pp. 241–252, Hayaishi, O., Ueda, K., eds. Academic Press, New York London

Download references

Author information

Correspondence to K. G. Wagner.

Additional information

This contribution is dedicated to Professor Augustin Betz on the occasion of his 65th birthday

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wagner, R., Feth, F. & Wagner, K.G. The pyridine-nucleotide cycle in tobacco. Planta 167, 226–232 (1986).

Download citation

Key words

  • NAD pyrophosphatase
  • Nicotinamidase
  • Nicotiana (pyridine-nucleotide cycle)
  • Nicotine biosynthesis
  • Pyridine nucleotide cycle