Purification and characterization of a new alpha-amylase of intermediate thermal stability from the yeast Lipomyces kononenkoae

A new alpha-amylase from the extracellular culture of the yeast Lipomyces kononenkoae CBS 5608 has been purified to homogeneity by ammonium sulphate treatment, affinity binding on cross-linked starch, and DEAE-Biogel A chromatography. The enzyme was monomeric, with an apparent M(r) of 76 kilodaltons...

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Published inBiochemistry and cell biology Vol. 73; no. 1-2; p. 41
Main Authors Prieto, J A, Bort, B R, Martínez, J, Randez-Gil, F, Buesa, C, Sanz, P
Format Journal Article
LanguageEnglish
Published Canada 01.01.1995
Subjects
alpha-Amylases - chemistry
alpha-Amylases - isolation & purification
alpha-Amylases - metabolism
Enzyme Stability
Glycosylation
Hydrogen-Ion Concentration
Isoelectric Point
Kinetics
Molecular Weight
Saccharomycetales - enzymology
Starch - metabolism
Temperature
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Summary:A new alpha-amylase from the extracellular culture of the yeast Lipomyces kononenkoae CBS 5608 has been purified to homogeneity by ammonium sulphate treatment, affinity binding on cross-linked starch, and DEAE-Biogel A chromatography. The enzyme was monomeric, with an apparent M(r) of 76 kilodaltons, pI < 3.5, and optimum pH 4.5-5.0, and exhibited intermediate thermal stability. The temperature for optimal enzyme activity was 70 degrees C. It is a glycoprotein with both N- and O-linked sugars. Kinetic analyses indicate that the enzyme has an endoamylolytic mechanism. The kM for soluble starch was 0.80 g.L-1 and the kcat was 622.s-1.
ISSN:0829-8211
DOI:10.1139/o95-005