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

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

doi:10.1007/s11274-004-3908-3

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 journal › Article › peer-review

5 Scopus citations

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 language	English (US)
Pages (from-to)	371-375
Number of pages	5
Journal	World Journal of Microbiology and Biotechnology
Volume	21
Issue number	3
DOIs	https://doi.org/10.1007/s11274-004-3908-3
State	Published - Apr 2005
Externally published	Yes

Keywords

Affinity adsorption
Bacillus licheniformis
α-amylase

ASJC Scopus subject areas

Biotechnology
Physiology
Applied Microbiology and Biotechnology

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10.1007/s11274-004-3908-3

Cite this

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title = "Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis",

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.",

keywords = "Affinity adsorption, Bacillus licheniformis, α-amylase",

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T1 - Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis

AU - Rao, M. Damodara

AU - Ratnam, B. V.V.

AU - VenkataRamesh, Dasari

AU - Ayyanna, C.

PY - 2005/4

Y1 - 2005/4

N2 - 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.

AB - 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.

KW - Affinity adsorption

KW - Bacillus licheniformis

KW - α-amylase

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

Abstract

Keywords

ASJC Scopus subject areas

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