Purification and characterization of the extracellular alpha-amylase from Clostridium acetobutylicum ATCC 824.

Applied and Environmental Microbiology
V PaquetP Soucaille

Abstract

The extracellular alpha-amylase (1,4-alpha-D-glucanglucanohydrolase; EC 3.2.1.1) from Clostridium acetobutylicum ATCC 824 was purified to homogeneity by anion-exchange chromatography (mono Q) and gel filtration (Superose 12). The enzyme had an isoelectric point of 4.7 and a molecular weight of 84,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was a monomeric protein, the 19-amino-acid N terminus of which displayed 42% homology with the Bacillus subtilis saccharifying alpha-amylase. The amino acid composition of the enzyme showed a high number of acidic and hydrophobic residues and only one cysteine residue per mole. The activity of the alpha-amylase was not stimulated by calcium ions (or other metal ions) or inhibited by EDTA, although the enzyme contained seven calcium atoms per molecule. alpha-Amylase activity on soluble starch was optimal at pH 5.6 and 45 degrees C. The alpha-amylase was stable at an acidic pH but very sensitive to thermal inactivation. It hydrolyzed soluble starch, with a Km of 3.6 g . liter-1 and a Kcat of 122 mol of reducing sugars . s-1 . mol-1. The alpha-amylase showed greater activity with high-molecular-weight substrates than with low-molecular-weight maltooligosacc...Continue Reading

References

Nov 1, 1976·Journal of Bacteriology·V BuonocoreA Gambacorta
Apr 1, 1973·Archives of Biochemistry and Biophysics·N Saito
Mar 1, 1970·Biotechnology and Bioengineering·M H Moseley, L Keay
Feb 1, 1966·Analytical Biochemistry·R R Barton
Jan 25, 1983·Nucleic Acids Research·M YangD Henner
May 1, 1959·Applied Microbiology·D SCOTT, L HEDRICK
Jun 1, 1952·Journal of Bacteriology·D SCOTT, L R HEDRICK
Aug 1, 1985·Applied and Environmental Microbiology·S F LeeL N Gibbins
Oct 1, 1985·Applied and Environmental Microbiology·S F LeeL N Gibbins
Dec 1, 1985·Applied and Environmental Microbiology·R De Mot, H Verachtert
Apr 1, 1987·Applied and Environmental Microbiology·S F LeeJ B Rattray
Apr 1, 1987·Applied and Environmental Microbiology·S F Lee, C W Forsberg
Jul 1, 1987·Applied and Environmental Microbiology·E MadiG Gottschalk
Jan 1, 1945·The Biochemical Journal·D J Hockenhull, D Herbert

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Citations

Jan 1, 1995·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·J A PrietoP Sanz
May 23, 2002·FEMS Microbiology Letters·Fabrice SabathéPhilippe Soucaille
May 11, 2005·Preparative Biochemistry & Biotechnology·Bilal Balkan, Figen Ertan
Jul 20, 2006·Applied Microbiology and Biotechnology·Dilli Rani NagarajanChandraraj Krishnan
Apr 2, 2008·Annals of the New York Academy of Sciences·Peter Dürre
Oct 10, 2007·Biotechnology Journal·Peter Dürre
Apr 13, 2007·The Journal of General and Applied Microbiology·Shunichi NakayamaKensuke Furukawa
Feb 28, 2019·Recent Patents on Biotechnology·Satya Eswari Jujjavarapu, Swasti Dhagat
May 18, 1999·Experimental Parasitology·A FerrerD Bout
May 1, 1995·Applied and Environmental Microbiology·N J Shih, R G Labbé

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