Cucurbita pepo (Pumpkin): In Vitro Production of Ascorbate Oxidase

  • M. Esaka
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 21)


Pumpkins (Cucurbita spp.), which belong to the family Cucurbitaceae, were originally found in Central and South America, and have long been cultivated throughout the world. The fruits, which contain a large amount of starch, proteins, free amino acids, and vitamins A, B, and C, have popularly been used as a vegetable, for pies, and for livestock feed. In Japan, the custom prevails of eating pumpkin fruits in the winter solstice, as they are thought to prevent a cold or palsy. The young vines and leafstalks have also been eaten as a vegetable. Dried ripe seeds (Cucurbitae semen) of cultivated varieties of Cucurbita pepo contain fixed oil, myosin, vitellin, and sugar, and have been used as food. Furthermore, the dried seeds have been used as a teniacide, since they contain resin (peporesin).


Diamine Oxidase Ascorbate Oxidase Cucurbita Pepo Ascorbic Acid Oxidase Barley Aleurone Layer 
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  1. Avigliano L, Vecchini P, Sirianni P, Marcozzi G, Marchesini A, Mandovi B (1983) A reinvestigation on the quaternary structure of ascorbate oxidase fromCucurbita pepo medullosa. Mol Biochem 56: 107–112Google Scholar
  2. Ayabe S, Yoshikawa T, Kobayashi M, Furuya T (1980) Biosynthesis of a retrochalcone echinatin: involvement of O-methyltransferase to licodione. Phytochemistry 19: 2331–2336CrossRefGoogle Scholar
  3. Bligny R, Douce R (1983) Excretion of laccase by sycamore (Acer pseudoplatanus L.) cells. Biochem J 209: 489–496PubMedGoogle Scholar
  4. Cheung WY (1980) Calmodulin plays a pivotal role in cellular regulation. Science 207: 19–27PubMedCrossRefGoogle Scholar
  5. Chibbar RN, Cella R, Albani D, Huystee RBV (1984) The growth of and peroxidase synthesis by two carrot cell lines. J Exp Bot 35: 1846–1852CrossRefGoogle Scholar
  6. Chrispeels M J, Varner JE (1967) Gibberellic acid-enhanced synthesis and release of a-amylase and ribonuclease by isolated barley aleurone layers. Plant Physiol 42: 398–406PubMedCrossRefGoogle Scholar
  7. Ciarrochi G, Cella R, Nielsen E (1981) Release of nucleotide cleaving and phosphatase from carrot cells grown in suspension culture. Physiol Plant 53: 357–377CrossRefGoogle Scholar
  8. Deikman J, Jones RL (1985) Control of a-amylase mRNA accumulation by gibberellic acid and calcium in barley aleurone layers. Plant Physiol 78: 192–198PubMedCrossRefGoogle Scholar
  9. Delhaize E, Dilworth MJ, Webb J (1986) The effects of copper nutrition and developmental state on the biosynthesis of diamine oxidase in clover leaves. Plant Physiol 82: 1126–1131PubMedCrossRefGoogle Scholar
  10. Esaka M, Asahi T (1982) Purification and properties of catalase from sweet potato root microbodies. Plant Cell Physiol 23: 315–322Google Scholar
  11. Esaka M, Suzuki K, Kubota K (1985) Determination method for L-ascorbic acid in foods with immobilized ascorbate oxidase. Agric Biol Chem 49: 2955–2960CrossRefGoogle Scholar
  12. Esaka M, Imagi J, Suzuki K, Kubota K (1988a) Formation of ascorbate oxidase in cultured pumpkin cells. Plant Cell Physiol 29: 231–235Google Scholar
  13. Esaka M, Uchida M, Fukui H, Kubota K, Suzuki K (1988b) Marked increase in ascorbate oxidase protein in pumpkin callus by adding copper. Plant Physiol 88: 656–660PubMedCrossRefGoogle Scholar
  14. Esaka M, Uchida M, Suzuki K, Kubota K (1989a) Increases in the activity and amount of ascorbate oxidase during the germination of pumpkin seeds. Agric Biol Chem 53: 1181–1182CrossRefGoogle Scholar
  15. Esaka M, Fukui H, Suzuki K, Kubota K (1989b) Secretion of ascorbate oxidase by suspension-cultured pumpkin cells. Phytochemistry 28: 117–119CrossRefGoogle Scholar
  16. Esaka M, Nishitani I, Fukui H, Suzuki K, Kubota K (1989c) Stimulation of ascorbate oxidase secretion from cultured pumpkin cells by divalent cations. Phytochemistry 28: 2655–2658CrossRefGoogle Scholar
  17. Esaka M, Suzuki K, Kubota K (1990a) Stimulation of ascorbate oxidase secretion from cultured pumpkin cells by Eosine Yellowish and potassium salicylate. Phytochemistry 29: 1547–1549CrossRefGoogle Scholar
  18. Esaka M, Hattori T, Fujisawa K, Sakajo S, Asahi T (1990b) Molecular cloning and nucleotide sequence of full-length cDNA for ascorbate oxidase from cultured pumpkin cells. Eur J Biochem 191: 537–541PubMedCrossRefGoogle Scholar
  19. Gardiner JD, Walker MD, Rottman A J (1980) Effect of A23187 on amylase release from dispersed acini prepared from guinea pig pancreas. Am J Physiol 238: 9457–9466Google Scholar
  20. Honda SI (1955) Ascorbic acid oxidase in barley roots. Plant Physiol 30: 174–181PubMedCrossRefGoogle Scholar
  21. Huystee RBV, Lobarzewski J (1982) An immunological study of peroxidase release by cultured peanut cells. Plant Sci Lett 27: 59–67CrossRefGoogle Scholar
  22. Ikeda T, Matsumoto T, Obi Y (1982) Influences of copper concentration on cytochrome aa3 formation and growth in cultured tobacco cells. Agric Biol Chem 46: 565–566CrossRefGoogle Scholar
  23. Jacobsen JV, Scandalios JG, Varner JE (1970) Multiple forms of amylase induced by gibberellic acid in isolated barley aleurone layers. Plant Physiol 45: 367–371PubMedCrossRefGoogle Scholar
  24. Jamieson JD, Palade JE (1977) Production of secretory proteins in animal cells. In: Brinkley BR, Porter KR (eds) International cell biology 1976–1977. Rockefeller Univ Press, New York, pp 308–317Google Scholar
  25. Jones RL, Carbonell J (1984) Regulation of the synthesis of barley aleurone a-amylase by gibberellic acid and calcium ions. Plant Physiol 76: 213–218PubMedCrossRefGoogle Scholar
  26. Jones RL, Jacobsen JV (1983) Calcium regulation of the secretion of a-amylase isozymes and other proteins from barley aleurone layers. Planta 158: 1–9CrossRefGoogle Scholar
  27. Kaulen H, Schell J, Kreuzaler F (1986) Light-induced expression of the chimeric chalcone synthase-NPTII gene in tobacco cells. EMBO J 5: 1–8PubMedGoogle Scholar
  28. Keyhani E, Keyhani J (1975) Cytochrome c oxidase biosynthesis and assembly in Candida utilis yeast cells. Arch Biochem Biophys 167: 596–602PubMedCrossRefGoogle Scholar
  29. Kornfeld R, Kornfeld S (1985) Assembly of asparagine-linked oligosacchrides. Annu Rev Biochem 54: 631–664PubMedCrossRefGoogle Scholar
  30. Lee MH, Dawson CR (1973) Ascorbate oxidase: further studies on the purification of the enzyme. J Biol Chem 248: 6596–6602PubMedGoogle Scholar
  31. Leube J, Grisebach H (1983) Further studies on induction of enzymes of phytoalexin synthesis in soybean and cultured soybean cells. Z Naturforsch 38: 730–735Google Scholar
  32. Mancini G, Carbonara AO, Heremans GF (1965) Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 2: 235–254PubMedCrossRefGoogle Scholar
  33. Matsumoto K, Yamada K, Osajima Y (1981) Ascorbate electrode for determination of L-ascorbic acid in food. An Chem 53: 1974–1979CrossRefGoogle Scholar
  34. Merchant S, Bogorad L (1987) Metal ion regulated gene expression: use of a plastocyanin-less mutant of Chlamydomonas reinhardtii to study the Cu(II)-dependent expression of cytochrome c-552. EMBO J 6: 2531–2535PubMedGoogle Scholar
  35. Mitsui T, Christeller JT, Hara-Nishimura I, Akazawa T (1984) Possible roles of calcium and calmodulin in the biosynthesis and secretion of a-amylase in rice seed scutellar epithelium. Plant Physiol 75: 21–25PubMedCrossRefGoogle Scholar
  36. Nakamura T, Makino N, Ogura Y (1968) Purification and properties of ascorbate oxidase from cucumber. J Biochem 64: 189–195PubMedGoogle Scholar
  37. Oberbacher MF, Vines HM (1963) Spectrophotometric assay of ascorbic acid oxidase. Nature 197: 1203–1204PubMedCrossRefGoogle Scholar
  38. Ouchterlony O, Nilsson LA (1973) Immunodiffusion and immunoelectrophoresis. In: Weir DM (ed) Handbook of experimental immunology. Blackwell, Oxford, 19, pp 1–39Google Scholar
  39. Polacco JC (1977) Nitrogen metabolism in soybean tissue culture: II. Urea utilization and urease synthesis require Ni2+. Plant Physiol 59: 827–830PubMedCrossRefGoogle Scholar
  40. Shatzman AR, Kosman DJ (1978) The utilization of copper and its role in the biosynthesis of copper-containing proteins in the fungus Dactylium dendroides. Biochim Biophys Acta 544: 163–179PubMedCrossRefGoogle Scholar
  41. Stark GR, Dawson CR (1962) On the accessibility of sulfhydryl groups in ascorbic acid oxidase. J Biol Chem 237: 712–716PubMedGoogle Scholar
  42. Takahashi N, Hotta T, Ishihara H, Mori M, Tejima S, Bligny R, Akazawa T, Endo S, Arata Y (1986) Xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells. Biochemistry 25: 388–395CrossRefGoogle Scholar
  43. Ueki K, Sato S (1971) Effect of inorganic phosphate on the extracellular acid phosphatase activity of tobacco cells cultured in vitro. Physiol Plant 24: 506–511CrossRefGoogle Scholar
  44. Vines HM, Oberbacher MF (1963) Citrus fruit enzyme: 1. Ascorbic acid oxidase in oranges. Plant Physiol 38: 333–337PubMedCrossRefGoogle Scholar
  45. Weis W (1975) Ascorbic acid and electron transport. Ann NY Acad Sci 258: 190–200PubMedCrossRefGoogle Scholar
  46. Wood PM (1978) Interchangeable copper and iron proteins in algal photosynthesis. Studies on plastocyanin and cytochrome c-552 in chlamydomonas. Eur J Biochem 87: 9–19PubMedCrossRefGoogle Scholar
  47. Yamauchi H, Yamawaki K, Ueda Y, Chachin K (1984) Subcellular localization of redox enzymes involving ascorbic acid in cucumber fruits. J Jpn Soc Hortic Sci 53: 347–353CrossRefGoogle Scholar
  48. Yamazaki Y, Konno H (1985) Three forms of a-glucosidase from suspension-cultured rice cells. Agric Biol Chem 49: 3383–3390CrossRefGoogle Scholar
  49. Zenk MH, Rueffer M, Amann M, Neumann BD, Nagakura N (1985) Benzylisoquinoline biosynthesis by cultivated plant cells and isolated enzymes. J Nat Prod 48: 725–738CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • M. Esaka
    • 1
  1. 1.Faculty of Applied Biological SciencesHiroshima UniversityKagamiyama, Higashi-Hiroshima 724Japan

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