Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1970 Oct;46(4):586–591. doi: 10.1104/pp.46.4.586

α-Amylase Isozymes in Gibberellic Acid-treated Barley Half-seeds

Y Tanaka a, T Akazawa a
PMCID: PMC396640  PMID: 16657510

Abstract

The presence of multiple forms of α-amylase in gibberellic acid-treated embryoless barley half-seeds was demonstrated by separation on diethylaminoethyl-Sephadex and isoelectric focusing polyacrylamide gel disc electrophoresis. Two major α-amylase fractions (A and B), each consisting of two to three isozyme components, were purified. α-Amylase fractions A and B were distinguishable in their reaction patterns. The optimal pH of fraction A α-amylase was found to reside in the acidic side (pH 5.0), as was determined by analyzing the reducing sugars formed as well as the paper chromatographic detection of reaction products. At neutral pH, 6.9, fraction A exhibited weak amylolytic activity in forming maltose. The α-amylase activity in fraction A was markedly stimulated by heat treatment (70 C/15 minutes). Fraction B, constituting a major part of amylases in the endosperm extract, was also found to be composed of α-amylase, as evidenced by the loss of enzyme activity upon allowing fractions A and B to stand at pH 3.3 for a prolonged period. The possible physiological function of the two different types of α-amylase in the carbohydrate breakdown of barley seeds is discussed.

Full text

PDF
586

Images in this article

587Image
on p.587
588Image
on p.588
589Image
on p.589

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akazawa T., Saio K., Sugiyama N. On the structural nature of fraction-I protein of rice leaves. Biochem Biophys Res Commun. 1965 Jul 12;20(2):114–119. doi: 10.1016/0006-291x(65)90332-3. [DOI] [PubMed] [Google Scholar]
  2. Chrispeels M. J., Varner J. E. Gibberellic Acid-enhanced synthesis and release of alpha-amylase and ribonuclease by isolated barley and aleurone layers. Plant Physiol. 1967 Mar;42(3):398–406. doi: 10.1104/pp.42.3.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chrispeels M. J., Varner J. E. Hormonal control of enzyme synthesis: on the mode of action of gibberellic Acid and abscisin in aleurone layers of barley. Plant Physiol. 1967 Jul;42(7):1008–1016. doi: 10.1104/pp.42.7.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Filner P., Varner J. E. A test for de novo synthesis of enzymes: density labeling with H2O18 of barley alpha-amylase induced by gibberellic acid. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1520–1526. doi: 10.1073/pnas.58.4.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jacobsen J. V., Scandalios J. G., Varner J. E. Multiple forms of amylase induced by gibberellic acid in isolated barley aleurone layers. Plant Physiol. 1970 Apr;45(4):367–371. doi: 10.1104/pp.45.4.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jacobsen J. V., Varner J. E. Gibberellic Acid-induced synthesis of protease by isolated aleurone layers of barley. Plant Physiol. 1967 Nov;42(11):1596–1600. doi: 10.1104/pp.42.11.1596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOYTER A., SCHRAMM M. The glycogen-amylase complex as a means of obtaining highly purified alpha-amylases. Biochim Biophys Acta. 1962 Dec 4;65:200–206. doi: 10.1016/0006-3002(62)91039-9. [DOI] [PubMed] [Google Scholar]
  9. Malacinski G. M., Rutter W. J. Multiple molecular forms of alpha-amylase from the rabbit. Biochemistry. 1969 Nov;8(11):4382–4390. doi: 10.1021/bi00839a024. [DOI] [PubMed] [Google Scholar]
  10. Matsumoto C., Sugiyama T., Akazawa T., Miyachi S. Structure and function of chloroplast proteins. IX. Further comparative studies on Chlorella and spinach leaf ribulose-1,5-diphosphate carboxylase. Arch Biochem Biophys. 1969 Dec;135(1):282–287. doi: 10.1016/0003-9861(69)90541-4. [DOI] [PubMed] [Google Scholar]
  11. Momotani Y., Kato J. Isozymes of alpha-Amylase Induced by Gibberellic Acid in Embryo-less Grains of Barley. Plant Physiol. 1966 Oct;41(8):1395–1396. doi: 10.1104/pp.41.8.1395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Murata T. Enzymic mechanism of starch breakdown in germinating rice seeds I. An analytical study. Plant Physiol. 1968 Dec;43(12):1899–1905. doi: 10.1104/pp.43.12.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Paleg L. G. Physiological Effects of Gibberellic Acid. II. On Starch Hydrolyzing Enzymes of Barley Endosperm. Plant Physiol. 1960 Nov;35(6):902–906. doi: 10.1104/pp.35.6.902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Paleg L. G. Physiological Effects of Gibberellic Acid: I. On Carbohydrate Metabolism and Amylase Activity of Barley Endosperm. Plant Physiol. 1960 May;35(3):293–299. doi: 10.1104/pp.35.3.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SHUSTER L., GIFFORD R. H. Changes in 3'-nucleotidase during the germination of wheatembryos. Arch Biochem Biophys. 1962 Mar;96:534–540. doi: 10.1016/0003-9861(62)90332-6. [DOI] [PubMed] [Google Scholar]
  16. Susor W. A., Kochman M., Rutter W. J. Heterogeneity of presumably homogeneous protein preparations. Science. 1969 Sep 19;165(3899):1260–1262. doi: 10.1126/science.165.3899.1260. [DOI] [PubMed] [Google Scholar]
  17. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  18. Varner J. E., Chandra G. R. HORMONAL CONTROL OF ENZYME SYNTHESIS IN BARLEY ENDOSPERM. Proc Natl Acad Sci U S A. 1964 Jul;52(1):100–106. doi: 10.1073/pnas.52.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Varner J. E. Gibberellic Acid Controlled Synthesis of alpha-Amylase in Barley Endosperm. Plant Physiol. 1964 May;39(3):413–415. doi: 10.1104/pp.39.3.413. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES