Isolation and characterization of an L-amino acid acylase fromAspergillus oryzae
- 32 Downloads
A scheme of isolating a highly purified L-amino acylase fromAspergillus oryzae is described which excludes extraction of the enzyme from the preparation “Amilorizin,” fractionation with ethanol, chromatography on DEAE-cellulose, and gel filtration through Sephadex G-200 and Bio-Gel P-300. The enzyme, as purified 1240-fold, has a molecular weight of 118,000, apparently consists of two subunits with a molecular weight of 60,000, is stable in the pH range of 7–10 and has an optimum pH of 8.9 and a pI of 4.0. Its amino acid composition has been determined and its substrate specificity has been studied. The acylase is a metalloenzyme: Co2+` ions in concentrations of 10−4–5·10−5 M increase the rate of hydrolysis of N-acetyl-L-amino acids three- to fourfold. It shows differences in its molecular and functional properties from acylase I obtained from porcine kidney.
KeywordsSodium Dodecyl Sulfate Amino Acid Composition Borate Buffer Porcine Kidney Nitr Obenzenediaz Onium
Unable to display preview. Download preview PDF.
- 2.E. N. Makarova, Biol. Zh. Arm.,32, No. 9, 850 (1979).Google Scholar
- 4.K. Hirano, I. Karube, and S. Suzuki, J. Ferment. Technol.,55, No. 4, 401 (1977).Google Scholar
- 5.Y. Kameda, T. Hase, S. Kanatomo, and K. Miyazaki, Chem. Pharm. Bull.,26, No. 9, 2710 (1978).Google Scholar
- 6.I. A. Yamskov, M. V. Budanov, L. S. Lobareva, M. K. Malinka, and V. D. Davankov, Biokhimiya,46, 651 (1981).Google Scholar
- 7.W. Kördel and Fr. Schneider, Z. Naturforsch.,32, 342 (1977).Google Scholar
- 8.Y. Frey, W. Kördel, and Fr. Schneider, Z. Naturforsch.,32, 759 (1977).Google Scholar
- 9.W. Kördel and Fr. Schneider, Z. Naturforsch.,32, 337 (1977).Google Scholar
- 10.V. K. Shvyadas, I. Yu. Galaev, N. A. Galstyan, and I. V. Berezin, Biokhimiya,45, 1361 (1980).Google Scholar
- 12.W. Kördel and Fr. Schneider, Z. Physiol. Chem.,356, 915 (1975).Google Scholar
- 13.F. Brubs and Ch. Schulze, Biochem. Z.,336, 162 (1962).Google Scholar
- 15.K. Waber and M. Osborn, J. Biol. Chem.,224, 4406 (1969).Google Scholar