Abstract
Vitamin C (L-ascorbic acid) has important antioxidant and metabolic functions in both plants and animals, but humans, and a few other animal species, have lost the capacity to synthesize it1. Plant-derived ascorbate is thus the major source of vitamin C in the human diet. Although the biosynthetic pathway of L-ascorbic acid in animals is well understood2, the plant pathway has remained unknown3—one of the few primary plant metabolic pathways forwhich this is the case. L-ascorbate is abundant in plants (found at concentrations of 1–5 mM in leaves and 25 mM in chloroplasts3,4) and may have roles in photosynthesis and transmembrane electron transport3,4,5. We found that D-mannose and L-galactose are efficient precursors for ascorbate synthesis and are interconverted by GDP-D-mannose-3,5-epimerase. We have identified an enzyme in pea and Arabidopsis thaliana, L-galactose dehydrogenase, that catalyses oxidation of L-galactose to L-galactono-1,4-lactone. We propose anascorbate biosynthesis pathway involving GDP-D-mannose, GDP-L-galactose, L-galactose and L-galactono-1,4-lactone, and have synthesized ascorbate from GDP-D-mannose by way of these intermediates in vitro. The definition of this biosynthetic pathway should allow engineering of plants for increased ascorbate production, thus increasing their nutritional value and stress tolerance.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Nishikimi, M., Fukuyama, R., Minoshima, S., Simizu, N. & Yagi, K. Cloning and chromosomal mapping of the human nonfunctional gene for L-gulono-γ-lactone oxidase, the enzyme for L-ascorbic and biosynthesis missing in man. J. Biol. Chem. 269, 13685–13688 (1994).
Burns, J. J. in Metabolic Pathways2nd edn Vol. 1 (ed. Greenberg, D. M.) 394–411 (Academic, New York, (1967)).
Smirnoff, N. The function and metabolism of ascorbic acid in plants. Ann. Bot. 78, 661–669 (1996).
Foyer, C. H. in Antioxidants in Higher Plants (eds Alscher, R. G. & Hess, J. L.) 31–58 (CRC, Boca Raton, (1993)).
Horemans, N., Asard, H. & Caubergs, R. J. The role of ascorbate free radical as an electron acceptor to cytochrome b -mediated transmembrane electron transport in higher plants. Plant Physiol. 104, 1455–1458 (1994).
Mapson, L. W. & Breslow, E. Biological synthesis of L-ascorbic acid: L-galactono-γ-lactone dehydrogenase. Biochem. J. 68, 395–406 (1958).
Oba, K., Ishikawa, S., Nishikawa, M., Mizuno, H. & Yamamoto, T. Purification and properties of L-galactono-γ-lactone dehydrogenase, a key enzyme for ascorbic acid biosynthesis, from sweet potato roots. J. Biochem. 117, 120–124 (1995).
Loewus, F. A. & Loewus, M. B. Biosynthesis and metabolism of L-ascorbic acid in plants. Crit. Rev. Plant Sci. 5, 101–119 (1987).
Mapson, L. W. & Isherwood, F. A. Biological synthesis of L-ascorbic acid: the conversion of derivatives of D-galacturonic acid to L-ascorbate in plant extracts. Biochem. J. 64, 13–22 (1956).
Loewus, F. A. Tracer studies of ascorbic acid formation in plants. Phytochemistry 2, 109–128 (1963).
Saito, K., Nick, J. A. & Loewus, F. A. D-Glucosone and L-sorbosone, putative intermediates of L-ascorbic acid biosynthesis in detected bean and spinach leaves. Plant Physiol. 94, 1496–1500 (1990).
Loewus, M. W., Bedgar, D. L., Saito, K. & Loewus, F. A. Conversion of L-sorbosone to L-ascorbic acid by a NADP+-dependent dehydrogenase in bean and spinach leaf. Plant Physiol. 94, 1492–1495 (1990).
Maier, E. & Kurtz, G. D-galactose dehydrogenase from Pseudomonas fluorescens. Methods Enzymol. 89, 176–181 (1982).
Schachter, H., Sarney, J., McGuire, E. J. & Roseman, S. Isolation of diphosphopyridine nucleotide-dependent L-fucose dehydrogenase from pork liver. J. Biol. Chem. 244, 4785–4792 (1969).
Conter, P. F., Guimarães, M. F. & Veiga, L. A. Induction and repression of L-fucose dehydrogenase of Pullularia pullulans. Can. J. Microbiol. 30, 753–757 (1984).
Kim, S.-T., Huh, W.-K., Kim, J-Y., Hwang, S-W. & Kang, S-O. D-Arabinose dehydrogenase and biosynthesis of erythroascorbate in Candida albicans. Biochim. Biophys. Acta 1297, 1–8 (1996).
Roberts, R. M. The metabolism of D-mannose-14-C to polysaccharide in corn roots. Specific labelling of L-galactose, D-mannose, and L-fucose. Arch. Biochem. Biophys. 145, 685–692 (1971).
Baydoun, E. A.-H. & Fry, S. C. [2-3H]Mannose incorporation in cultured plant cells: investigation of L-galactose residues of the primary wall. J. Plant Physiol. 132, 484–490 (1988).
Barber, G. A. Observations on the mechanisms of the reversible epimerization of GDP-D-mannose to GDP-L-galactose by an enzyme from Chlorella pyrenidosa. J. Biol. Chem. 254, 7600–7603 (1979).
Barber, G. A. Synthesis of L-galactose by plant enzyme systems. Arch. Biochem. Biophys. 147, 619–623 (1971).
1. Feingold, D. S. in Encyclopedia of Plant Physiology, Vol. 13A (eds Loewus, F. A. & Tanner, W.) 3–76 (Springer, Berlin, (1982)).
Roberts, R. M. & Harrer, E. Determination of L-galactose in polysaccharide material. Phytochemistry 12, 2679–2682 (1973).
Harris, G. C. et al. Mannose metabolism in corn and its impact on leaf metabolites, photosynthetic gas exchange, and chlorophyll fluorescence. Plant Physiol. 82, 1081–1089 (1986).
Conklin, P. L., Williams, E. H. & Last, R. L. Environmental stress sensitivity of an ascorbic acid-deficient Arabidopsis mutant. Proc. Natl Acad. Sci. USA 93, 9970–9974 (1996).
Conklin, P. L., Pallanca, J. E., Last, R. L. & Smirnoff, N. L-Ascorbic acid metabolism in the ascorbate-deficient Arabidopsis mutant vtc1. Plant Physiol. 115, 1277–1285 (1997).
Chen, Y-T., Isherwood, F. A. & Mapson, L. W. Quantitative estimation of ascorbic acid and related substances in biological extracts by separation on a paper chromatogram. Biochem. J. 55, 821–823 (1953).
Andrews, M. A. Capillary gas-chromatographic analysis of monosaccharides: improvements and comparisons using trifluoroacetylation and trimethylsilylation of sugar O-benzyl and O-methyl oximes. Carbohydrate Res. 194, 1–19 (1989).
Ghebragzabher, M., Ruffini, S., Monaldi, B. & Lato, M. Thin layer chromatography of monosaccharides. J. Chromatogr. 127, 133–162 (1976).
Dawson, R. M. C., Elliott, D. C., Elliott, W. H. & Jones, K. M. (eds) Data for Biochemical Research2nd edn. (Clarendon, Oxford, (1969)).
Acknowledgements
The research was supported by a BBSRC studentship (G.L.W.), a Nuffield Student Bursary (M.A.J.) and Zeneca Agrochemicals. We thank Nippon-Roche for the gift of L-sorbosone. Earlier work by J. Pallanca, funded by the BBSRC BOMRIP programme, provided a basis for this research. We thank M. Raymond for technical assistance; J.Kingdon and J. Hindley for growing the plants; and W. Schuch for his support.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wheeler, G., Jones, M. & Smirnoff, N. The biosynthetic pathway of vitamin C in higher plants. Nature 393, 365–369 (1998). https://doi.org/10.1038/30728
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/30728
This article is cited by
-
Integrated metabolome and transcriptome analysis of differences in quality of ripe Lycium barbarum L. fruits harvested at different periods
BMC Plant Biology (2024)
-
Hydrogen-based irrigation increases yield and improves quality of Chinese cabbage by enhancing nutrient composition and antioxidant capabilities
Horticulture, Environment, and Biotechnology (2024)
-
Transcriptome comparison analyses in UV-B induced AsA accumulation of Lactuca sativa L
BMC Genomics (2023)
-
Ascorbate oxidation stimulates rice root growth via effects on auxin and abscisic acid levels
Plant Growth Regulation (2023)
-
Hydrogen Peroxide Induced Antioxidant-Coupled Redox Regulation of Germination in Rice: Redox Metabolic, Transcriptomic and Proteomic Evidences
Journal of Plant Growth Regulation (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.