Skip to main content
SpringerLink
Log in

Precepitation and purification of amylase enzyme produced byStreptomyces aureofaciens 77

  • Original Articles
  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

Precipitation and purification of amylase secreted byStreptomyces aureofaciens 77 in liquid inorganic salts-starch medium under the optimum conditions were carried out. Ammonium sulphate fractionation was used to precipitate amylase in cell free culture filtrate. (NH4)2SO4 at a concentration of 50–70% saturation gave the highest enzyme yield. The obtained precipitates were redissolved in phosphate buffer (pH 7.0) and subjected to dialysis. The dialyzed enzyme preparation was applied to DEAE-cellulose column chromatography which resulted in an increase of purification up to 59.48 fold. A further step of purification was done by applying the obtained puified sample to Sephadex-G200 column chromatography which resulted in an increase of purification up to 73.92 fold. The results clearly indicated that the isolated amylase from S.aureofaciens 77 was only on type.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature Cited

  1. Price, N.C. and Stevens, L.: Fundamentals of enzymology. Oxford Univ. Press, Oxford, New York (1982).

    Google Scholar 

  2. Grootegeed, J.A., Lauwers, A. and Heinen, W.: Separation and partial purification of extracellular amylase and protonase from B.caldohyticus.Arch. Microbiol.,90, 223 (1973).

    Google Scholar 

  3. Upton, Mary E. and Fogarty, W.M.: Production and purification of thermostable amylase and protease ofThermomonspora viridis.Appl. Environ. Microbiol. 33(1), 59 (1976).

    Google Scholar 

  4. Ghai, S.K. and Chahal, D.S.: Alpha-amylase fromAspergillus flavus. Production, purification and properties.Indian J. Agric. Res. 14, 146 (1980).

    Google Scholar 

  5. Obi, S.K.C. and Odibo, F.J.C. Partial purification and characterization of thermostable Actinomycete β-amylase.Appl. Environ. Microbiol.47, 571 (1984).

    PubMed  CAS  Google Scholar 

  6. Himmelhoch, S.R.:Methods Enzymol. 22, 273 (1971).

    Google Scholar 

  7. Fischer, L.: An introduction to gel chromatography, North-Holand, Amesterdam (1969).

    Google Scholar 

  8. Reiland, J.:Methods Enzymol. 22, 287 (1971).

    Article  Google Scholar 

  9. Smith, B. and Roe, J.: Colorimetric determination of amylase activity.J. Biol. Chem. 227, 357 (1957).

    PubMed  CAS  Google Scholar 

  10. Caraway, W.T.: A stable starch substrate for the determination of amylase in serum and other body fluids.Am. J. Clin. Pathol. 32, 97 (1959).

    PubMed  CAS  Google Scholar 

  11. Lowry, O.H., Rosebrough, N., Farr, A. and Randall, R.: Protein measurement with the folin phenol reagent.AJ. Biol. Chem. 193, 265 (1951).

    CAS  Google Scholar 

  12. Refaat, A.A.: Studies on the production of microbial amylase. Ph.D. Thesis, Faculty of Agric., Ain Shams Univ., (1978).

  13. Ibrahim, M.M.K.: Production of some enzymes by Streptomyces. Ph.D. Thesis, Faculty of Agric. Al-Azhar University, 1983.

  14. Safwat, M.S.A., Mahmoud, S.A.Z., Abdel-Nasser, M. and Attia, R.M.: Isolation and purification of bacterial alpha-amylase.Zentrable. Milkrobiol. 138(3), 247 (1983).

    CAS  Google Scholar 

  15. De Mot, R. and Verachtert, H.: Purification and characterization of extracellular amylolytic enzymes from the yeastFilobasiduim capsuligenum.Appl. Environ. Microbiol.,48, 1474 (1985).

    Google Scholar 

  16. Shatta, A.M., Ibrahim, A.N., El-Kousy, S.M. and Ibrahim, M.M.K.: Proteinase production byStreptomyces griseus 550. III-Precipitation and purification of proteinase.Al-Azhar. J. Agric. Res. 3, (1986).

Download references

Author information

Authors and Affiliations

  1. Botany Department, Faculty of Agriculture, Al-Azhar University, Madinet Nasr, Egypt

    A. N. Ibrahim

  2. Mycology Department, National Organization for Drug Control and Research, Egypt

    F. H. Ahmed, M. M. K. Ibrahim & M. A. I. Arafa

  3. Faculty of Home Economic, Hellwan University, Cairo, Egypt

    A. N. Ibrahim, F. H. Ahmed, M. M. K. Ibrahim & M. A. I. Arafa

Authors
  1. A. N. Ibrahim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. H. Ahmed
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. M. K. Ibrahim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. A. I. Arafa
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ibrahim, A.N., Ahmed, F.H., Ibrahim, M.M.K. et al. Precepitation and purification of amylase enzyme produced byStreptomyces aureofaciens 77 . Arch. Pharm. Res. 13, 28–32 (1990). https://doi.org/10.1007/BF02857830

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02857830

Keywords

Search

Navigation