Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1984 May 1;219(3):849–855. doi: 10.1042/bj2190849

Glycosidases induced in Aspergillus tamarii. Mycelial alpha-D-galactosidases.

A Civas, R Eberhard, P Le Dizet, F Petek
PMCID: PMC1153553  PMID: 6331398

Abstract

Two alpha-D-galactosidases (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) produced by Aspergillus tamarii were purified from the mycelial extract by a procedure including chromatography on hydroxyapatite, DEAE-cellulose and ECTEOLA-cellulose. Each of these enzymes showed a single protein band corresponding to the alpha-D-galactosidase activity when examined by polyacrylamide-gel electrophoresis. They catalysed the hydrolysis of o-nitrophenyl alpha-D-galactoside, melibiose, raffinose and stachyose, but did not attack the galactomannans. Their Mr values were respectively 265000 +/- 5000 and 254000 +/- 5000 by the method of Hedrick & Smith [(1968) Arch. Biochem. Biophys. 126, 155-164]. Polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate in each case showed a single protein band, with Mr 88000 and 77500 respectively. The purified enzymes contained carbohydrate, consisting of N-acetylglucosamine, mannose, glucose and galactose in the estimated molar proportions of 1:9:5:8 in alpha-galactosidase I.

Full text

PDF
849

Images in this article

852Fig. 1.
on p.852

Selected References

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

  1. Adya S., Elbein A. D. Glycoprotein enzymes secreted by Aspergillus niger: purification and properties of alpha-glaactosidase. J Bacteriol. 1977 Feb;129(2):850–856. doi: 10.1128/jb.129.2.850-856.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bahl O. P., Agrawal K. M. Glycosidases of Aspergillus niger. I. Purification and characterization of alpha- and beta-galactosidases and beta-N-acetylglucosaminidase. J Biol Chem. 1969 Jun 10;244(11):2970–2978. [PubMed] [Google Scholar]
  3. Bouquelet S., Spik G., Montreuil J. Properties of a beta-D-mannosidase from Aspergillus niger. Biochim Biophys Acta. 1978 Feb 10;522(2):521–530. doi: 10.1016/0005-2744(78)90084-0. [DOI] [PubMed] [Google Scholar]
  4. COURTOIS J. E., LE DIZET P., WICKSTROM A. Recherches sur les galactosides du Lychnis dioica. III. Etude des racines récoltées en avril: isolement d'un heptasaccharide. Bull Soc Chim Biol (Paris) 1958;40(7-8):1059–1065. [PubMed] [Google Scholar]
  5. Elbein A. D., Adya S., Lee Y. C. Purification and properties of a beta-mannosidase from Aspergillus niger. J Biol Chem. 1977 Mar 25;252(6):2026–2031. [PubMed] [Google Scholar]
  6. Eylar E. H. On the biological role of glycoproteins. J Theor Biol. 1966 Jan;10(1):89–113. doi: 10.1016/0022-5193(66)90179-2. [DOI] [PubMed] [Google Scholar]
  7. HJERTEN S., LEVIN O., TISELIUS A. Protein chromatography on calcium phosphate columns. Arch Biochem Biophys. 1956 Nov;65(1):132–155. doi: 10.1016/0003-9861(56)90183-7. [DOI] [PubMed] [Google Scholar]
  8. Hedrick J. L., Smith A. J. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968 Jul;126(1):155–164. doi: 10.1016/0003-9861(68)90569-9. [DOI] [PubMed] [Google Scholar]
  9. Kapitany R. A., Zebrowski E. J. A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal Biochem. 1973 Dec;56(2):361–369. doi: 10.1016/0003-2697(73)90202-9. [DOI] [PubMed] [Google Scholar]
  10. Keller R. K., Swank G. D. Tunicamycin does not block ovalbumin secretion in the oviduct. Biochem Biophys Res Commun. 1978 Nov 29;85(2):762–768. doi: 10.1016/0006-291x(78)91226-3. [DOI] [PubMed] [Google Scholar]
  11. Kuo S. C., Lampen J. O. Tunicamycin--an inhibitor of yeast glycoprotein synthesis. Biochem Biophys Res Commun. 1974 May 7;58(1):287–295. doi: 10.1016/0006-291x(74)90925-5. [DOI] [PubMed] [Google Scholar]
  12. Lineback D. R., Russell I. J., Rasmussen C. Two forms of the glucoamylase of Aspergillus niger. Arch Biochem Biophys. 1969 Nov;134(2):539–553. doi: 10.1016/0003-9861(69)90316-6. [DOI] [PubMed] [Google Scholar]
  13. Loh Y. P., Gainer H. The role of glycosylation on the biosynthesis, degradation, and secretion of the ACTH-beta-lipotropin common precursor and its peptide products. FEBS Lett. 1978 Dec 15;96(2):269–272. doi: 10.1016/0014-5793(78)80415-3. [DOI] [PubMed] [Google Scholar]
  14. MAIZEL J. V., Jr PREPARATIVE ELECTROPHORESIS OF PROTEINS IN ACRYLAMIDE GELS. Ann N Y Acad Sci. 1964 Dec 28;121:382–390. doi: 10.1111/j.1749-6632.1964.tb14211.x. [DOI] [PubMed] [Google Scholar]
  15. Mizrahi A., O'Malley J. A., Carter W. A., Takatsuki A., Tamura G., Sulkowski E. Glycosylation of interferons. Effects of tunicamycin on human immune interferon. J Biol Chem. 1978 Nov 10;253(21):7612–7615. [PubMed] [Google Scholar]
  16. Pazur J. H., Knull H. R., Cepure A. Glycoenzymes: structure and properties of the two forms of glucoamylase from Aspergillus niger. Carbohydr Res. 1971 Nov;20(1):83–96. doi: 10.1016/s0008-6215(00)84951-4. [DOI] [PubMed] [Google Scholar]
  17. Pazur J. H., Knull H. R., Simpson D. L. Glycoenzymes: a note on the role for the carbohydrate moieties. Biochem Biophys Res Commun. 1970 Jul 13;40(1):110–116. doi: 10.1016/0006-291x(70)91053-3. [DOI] [PubMed] [Google Scholar]
  18. Rudick M. J., Elbein A. D. Glycoprotein enzymes secreted by Aspergillus fumigatus. Purification and properties of beta-glucosidase. J Biol Chem. 1973 Sep 25;248(18):6506–6513. [PubMed] [Google Scholar]
  19. Rudick M. J., Elbein A. D. Glycoprotein enzymes secreted by Aspergillus fumigatus: purification and properties of a second beta-glucosidase. J Bacteriol. 1975 Oct;124(1):534–541. doi: 10.1128/jb.124.1.534-541.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rudick M. J., Fitzgerald Z. E., Rudick V. L. Intra- and extracellular forms of alpha-glucosidase from Aspergillus niger. Arch Biochem Biophys. 1979 Apr 1;193(2):509–520. doi: 10.1016/0003-9861(79)90058-4. [DOI] [PubMed] [Google Scholar]
  21. Sedmak J. J., Grossberg S. E. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. doi: 10.1016/0003-2697(77)90428-6. [DOI] [PubMed] [Google Scholar]
  22. Shimada Y., Shinmyo A., Enatsu T. Purification and properties of one component of acid phosphatase produced by Aspergillus niger. Biochim Biophys Acta. 1977 Feb 9;480(2):417–427. doi: 10.1016/0005-2744(77)90034-1. [DOI] [PubMed] [Google Scholar]
  23. Struck D. K., Siuta P. B., Lane M. D., Lennarz W. J. Effect of tunicamycin on the secretion of serum proteins by primary cultures of rat and chick hepatocytes. Studies on transferrin, very low density lipoprotein, and serum albumin. J Biol Chem. 1978 Aug 10;253(15):5332–5337. [PubMed] [Google Scholar]
  24. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  25. Widmer F., Leuba J. L. beta-Galactosidase from Aspergillus niger. Separation and characterization of three multiple forms. Eur J Biochem. 1979 Oct 15;100(2):559–567. doi: 10.1111/j.1432-1033.1979.tb04202.x. [DOI] [PubMed] [Google Scholar]
  26. Williams J., Villarroya H., Petek F. alpha-Galactosidases II, III and IV from seeds of Trifolium repens. Purification, physicochemical properties and mode of galactomannan hydrolysis in vitro. Biochem J. 1978 Dec 1;175(3):1069–1077. doi: 10.1042/bj1751069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zanetta J. P., Breckenridge W. C., Vincendon G. Analysis of monosaccharides by gas-liquid chromatography of the O-methyl glycosides as trifluoroacetate derivatives. Application to glycoproteins and glycolipids. J Chromatogr. 1972 Jul 5;69(2):291–304. doi: 10.1016/s0021-9673(00)92897-8. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES