1887

Abstract

Two extracellular glucoamylases (EC 3.2.1.3), glucoamylase P and glucoamylase S, were purified to homogeneity from the culture medium of (ATCC 20495; formerly ) by a new method. Their apparent molecular masses (71 kDa glucoamylase P; 78 kDa glucoamylase S) and catalytic properties agreed well with those previously reported in the literature. Heat inactivation studies suggested that the high debranching (1,6-glycosidic) activity of glucoamylase P preparations (measured with pullulan) may reside in the same protein molecule as its 1,4-glycosidic activity (measured with soluble starch). Although glucoamylase S had virtually no debranching activity, it cross-reacted with polyclonal antibodies raised against glucoamylase P, and the two enzymes had very similar amino acid compositions. However, peptide mapping and amino-terminal sequencing studies of the peptides showed that the two enzymes have different sequences and must be encoded by different genes.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-5-913
1990-05-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/5/mic-136-5-913.html?itemId=/content/journal/micro/10.1099/00221287-136-5-913&mimeType=html&fmt=ahah

References

  1. Axén R., Porath J., Ernback S. 1967; Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature; London: 2141302–1304
    [Google Scholar]
  2. Boel E., Hjort I., Svensson B., Norris F., Norris K.E., Fiil N.P. 1984; Glucoamylases G1 and G2 from Aspergillus niger are synthesized from two different but closely related mRNAs. EMBO Journal 3:1097–1102
    [Google Scholar]
  3. Bradford M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  4. Chang J.Y., Knecht R., Braun D;G. 1983; Amino acid analysis in the picomole range by precolumn derivatization and high-performance liquid chromatography. Methods in Enzymology 91:41–48
    [Google Scholar]
  5. Einarsson S., Josefsson B., Lagerkvist S. 1983; Determination of amino acids with 9-fluorenylmethyl chloroformate and reversed-phase high-performance liquid chromatography. Journal of Chromatography 282:609–618
    [Google Scholar]
  6. Friedman M., Krull L.H., Cavins J.F. 1970; Reactions of amino acids, peptides and proteins with α,β-unsaturated compounds. XIV. Chromatographic determination of cystine and cysteine residues in proteins as S-β-(4-pyridylethyl)cysteine. Journal of Biological Chemistry 245:3868–3871
    [Google Scholar]
  7. Fullmer C.S. 1984; Identification of cysteins-containing peptides in protein digests by high-performance liquid chromatography. Analytical Biochemistry 142:336–339
    [Google Scholar]
  8. Hayashida S., Nakahara K., Kuroda K., Kamachi T., Ohta K., Iwanaga S., Miyata T., Sakaki Y. 1988; Evidence for posttranslational generation of multiple forms of Aspergillus awamori glucoamylase. Agricultural and Biological Chemistry 52:273–275
    [Google Scholar]
  9. Itoh T., Ohtsuki I., Yamashita I., Fukui S. 1987; Nucleotide sequence of the glucoamylase gene GLU1 in the yeast Saccharo-mycopsis fibuligera. Journal of Bacteriology 169:4171–4176
    [Google Scholar]
  10. Kalkkinen N., Tilgmann C. 1988; A gas-pulsed-liquid-phase sequencer constructed from a Beckman 890D instrument by using Applied Biosystems delivery and cartridge blocks. Journal of Protein Chemistry 7:242–243
    [Google Scholar]
  11. Kennedy J.F., Cabalda V.M., White C.A. 1988; Enzymic starch utilization and genetic engineering. Trends in Biotechnology 6:184–189
    [Google Scholar]
  12. Labout J.J.M. 1985; Conversion of liquefied starch into glucose using a novel glucoamylase system. Starch/Starke 37:157–161
    [Google Scholar]
  13. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  14. Lineback D.R., Bauman W.E. 1970; Properties of a glucoamylase from Aspergillus phoenicis. Carbohydrate Research 14:341–353
    [Google Scholar]
  15. Manjunath P., Shenoy B.C., Raghavendrarao M.R. 1983; Fungal glucoamylases. Journal of Applied Biochemistry 5:235–260
    [Google Scholar]
  16. Mccleary B.V., Anderson M.A. 1980; Hydrolysis of α-d-glucans and α-d-gluco-oligosaccharides by Cladosporium resinae glucoamylases. Carbohydrate Research 86:77–96
    [Google Scholar]
  17. Ono K., Shintani K., Shigeta S., Oka S. 1988; Various molecular species in glucoamylase from Aspergillus niger. Agricultural and Biological Chemistry 52:1689–1698
    [Google Scholar]
  18. Penke B., Ferenczi R., Kovacs K. 1974; A new acid hydrolysis method for determining tryptophan in peptides and proteins. Analytical Biochemistry 60:45–50
    [Google Scholar]
  19. Reeck G. 1987; Amino acid compositions of selected proteins. In CRC Handbook of Biochemistry and Molecular Biology 3 Proteins , 3rd edn. pp. 504–519 Fasman G.D. Edited by Boca Raton, Florida: CRC Press;
    [Google Scholar]
  20. Svensson B., Larsen K., Gunnarsson A. 1986; Characterization of a glucoamylase G2 from Aspergillus niger. European Journal of Biochemistry 154:497–502
    [Google Scholar]
  21. Takahashi T., Inokushi N., Irie M. 1978; Purification and some properties of three forms of glucoamylase from a Rhizopus species. Journal of Biochemistry 84:1183–1194
    [Google Scholar]
  22. Takahashi T., Inokushi N., Irie M. 1981; Purification and characterization of a glucoamylase from Aspergillus saitoi. Journal of Biochemistry 89:125–134
    [Google Scholar]
  23. Tanaka Y., Ashikari T., Nakamura N., Kiuchi N., Shibano Y., Amachi T., Yoshizumi H. 1986; Comparison of amino acid sequences of three glucoamylases and their structure-function relationships. Agricultural and Biological Chemistry 50:965–969
    [Google Scholar]
  24. Tsuboi A., Yamasaki Y., Suzuki Y. 1974; Two forms of glucoamylase from Mucor rouxianus. 1. Purification and crystallization. Agricultural and Biological Chemistry 38:543–550
    [Google Scholar]
  25. Yamasaki Y., Suzuki Y., Ozawa J. 1977; Purification and properties of two forms of glucoamylase from Penicillium oxalicum. Agricultural and Biological Chemistry 41:755–762
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-5-913
Loading
/content/journal/micro/10.1099/00221287-136-5-913
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error