1887

Abstract

Activity assays based on selective product detection were used to identify the classes of amylase present in culture filtrates of the phytopathogen When transferred from a glucose medium to a medium containing starch as sole carbohydrate, secreted amylolytic activity into liquid shake cultures starting 2-3 d after inoculation, and produced levels of activity up to 0.45 IU ml (assayed by the -hydroxybenzoic acid hydrazide — PAHBAH — reducing sugar method), about 20 times higher than the activity in comparable glucose-grown controls, suggesting that starch (or a starch degradation product) was required to induce amylase synthesis. In four separate experiments, the ratio of total reducing sugar production (assayed by the PAHBAH method) to glucose production (assayed by the glucose-specific glucose oxidase — GO — method) was about 1.0 in most samples of culture filtrate, clearly demonstrating that the major amylolytic enzyme secreted by is a glucoamylase (exo-1,4-a-d-glucan glucanohydrolase, EC 3.2.1.3, synonym amylogluco-sidase). On one occasion only (a 7 d sample in one experiment) was evidence obtained for a-amylase (endo-1,4-a-d-glucan glucanohydrolase, EC 3.2.1.1) also being present in the culture filtrate (PAHBAH/GO ratio 1.37); the presence of a-amylase in this sample was confirmed by native polyacrylamide gel electrophoresis using a starch-iodine activity stain.

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1991-10-01
2024-03-29
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References

  1. Abe J.-I., Nakajima K., Nagano H., Hizukuri S., Obata K. 1988; Properties of the raw-starch digesting amylase of Aspergillus sp. K-27 : a synergistic action of glucoamylase and alpha-amylase. Carbohydrate Research 174:85–92
    [Google Scholar]
  2. Ashikari T., Nakamura N., Tanaka Y., Kiuchi N., Shibano Y., Tanaka T., Amachi T., Yoshizumi H. 1986; Rhizopus raw-starch-degrading glucoamylase : its cloning and expression in yeast. Agricultural and Biological Chemistry 50:957–964
    [Google Scholar]
  3. Blakeney A. B., Mutton L. L. 1980; A simple colorimetrie method for the determination of sugars in fruit and vegetables. Journal of the Science of Food and Agriculture 31:889–897
    [Google Scholar]
  4. Boel E., Svensson H., Norris K., Fhl N. 1984; Glucoamylases G1 and G2 from Aspergillus niger are synthesised from two different but closely related mRNAs. EMBO Journal 3:1097–1102
    [Google Scholar]
  5. Charya M., Reddy S., Kumar B. 1983; α-Amylase production in relation to assimilation of starch by three fungi. Proceedings of the National Academy of Sciences of India 53:19–25
    [Google Scholar]
  6. De Mot R., Verachtert H. 1986; Enhanced production of extracellular α-amylase and glucoamylase by amylolytic yeasts using β-cyclodextrin as carbon source. Applied Microbiology and Biotechnology 24:459–462
    [Google Scholar]
  7. Gibson R. A., Paleg L. G. 1975; Further experiments on the oc-amylase-containing lysosomes of wheal aleurone cells. Australian Journal of Plant Physiology 2:41–49
    [Google Scholar]
  8. Irwin J. A. G., Cameron D. F. 1978; Two diseases of Stylosanthes spp. caused by Colletotrichum gloeosporioides in Australia, and pathogenic specialization within one of the causal organisms. Australian Journal of Agricultural Research 29:305–317
    [Google Scholar]
  9. Jodal I., Kandra L., Harangi J., Nanasi P., Szejtli J. 1984; Hydrolysis of cyclodextrin by Aspergillus oryzae α-amylase. Starch 36:140–143
    [Google Scholar]
  10. Kakefuda G., Duke S. 1984; Electrophoretic transfer as a technique for the detection and identification of plant amylolytic enzymes in polyacryamide gels. Plant Physiology 75:278–280
    [Google Scholar]
  11. Lever M. 1972; A new reaction for colorimetrie determination of carbohydrates. Analytical Biochemistry 47:273–279
    [Google Scholar]
  12. McKay D. B., Tanner G. P., Maclean D. J., Scott K. J. 1987; Detection of polyols and sugars by cuprammonium ion in the presence of strong base. Analytical Biochemistry 165:392–398
    [Google Scholar]
  13. Montenecourt B., EvElelgh D. 1985; Fungal carbohydrases : amylases and cellulases. Gene Manipulations in Fungi491–512 Bennet J. W., Lasure L. L. New York & London: Academic Press;
    [Google Scholar]
  14. Nunberg J., Meade J., Cole G., Lawyer F., McCabe P., Schweikart V., Tal R., Wittman V., Flatgaard J., Innis M. 1984; Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori . Molecular and Cellular Biology 4:2306–2315
    [Google Scholar]
  15. Sierks M. R., Ford C, Reilly P. J., Svensson B. 1989; Site-directed mutagenesis at the active site Trpl20 of Aspergillus awamori glucoamylase. Protein Engineering 2:621–625
    [Google Scholar]
  16. Slade S. J., Harris R. F., Smith C. S., Andrews J. H. 1987; Microcycle conidiation and spore-carrying capacity of Colletotrichum gloeosporioides on solid media. Applied and Environmental Microbiology 53:2106–2110
    [Google Scholar]
  17. Smith B. W., Roe J. H. 1949; A photometric method for the determination of α-amylase in blood and urine, with use of the starch-iodine colour. Journal of Biological Chemistry 179:53–59
    [Google Scholar]
  18. Trinder P. 1969; Determination of blood glucose using glucose oxidase with an alternative oxygen acceptor. Annals of Clinical Biochemistry 6:24–27
    [Google Scholar]
  19. Ueda S. 1981; Fungal glucoamylases and raw starch digestion. Trends in Biochemical Sciences 6:89–90
    [Google Scholar]
  20. Williams A. M., Maclean D. J., Scott K. J. 1984; Cellular location and properties of invertase in mycelium of Puccinia graminis . New Phytologist 98:451–463
    [Google Scholar]
  21. Yamashita I., Suzuki K., Fukui S. 1985; Nucleotide sequence of the extracellular glucoamylase gene STA 1 in the yeast Saccharomyces diastaticus . Journal of Bacteriology 161:567–573
    [Google Scholar]
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