Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis

M. Damodara Rao, B. V V Ratnam, Dasari VenkataRamesh, C. Ayyanna

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

A rapid and efficient method the exploiting affinity of α-amylase for its substrate starch is described. α-amylase from Bacillus licheniformis was purified to homogeneity by ammonium sulphate precipitation and affinity chromatography with 230-fold purification. The α-amylase adsorption to various starches was examined in order to screen its ability for highest binding to starch. The α-amylase was bound to starch more tenaciously, hence various eluants like maltose, soluble starch and high salts could not elute the bound α-amylase. However, the bound α-amylase was instantly eluted using 2% (w/v) dextrin. The purified enzyme showed a single polypeptide on SDS-PAGE, with a molecular weight of 58 kD. Western blot analysis confirmed the specificity of antibody raised against purified α-amylase.

Original languageEnglish (US)
Pages (from-to)371-375
Number of pages5
JournalWorld Journal of Microbiology and Biotechnology
Volume21
Issue number3
DOIs
StatePublished - Apr 2005
Externally publishedYes

Fingerprint

Bacillus licheniformis
Bacilli
Amylases
amylases
rapid methods
Purification
Starch
starch
Affinity chromatography
dextrins
Maltose
Antibody Specificity
Ammonium Sulfate
affinity chromatography
maltose
Affinity Chromatography
ammonium sulfate
Adsorption
Polyacrylamide Gel Electrophoresis
polypeptides

Keywords

  • α-amylase
  • Affinity adsorption
  • Bacillus licheniformis

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Plant Science
  • Biochemistry
  • Food Science
  • Microbiology
  • Biotechnology

Cite this

Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis. / Rao, M. Damodara; Ratnam, B. V V; VenkataRamesh, Dasari; Ayyanna, C.

In: World Journal of Microbiology and Biotechnology, Vol. 21, No. 3, 04.2005, p. 371-375.

Research output: Contribution to journalArticle

Rao, M. Damodara ; Ratnam, B. V V ; VenkataRamesh, Dasari ; Ayyanna, C. / Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis. In: World Journal of Microbiology and Biotechnology. 2005 ; Vol. 21, No. 3. pp. 371-375.
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