Purification and Characterization ofα-Glucosidase from Torulaspora pretoriensis YK-1

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概要

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• α-glucosidase was partially purified 103-fold from a cell-free extract of Torulaspora pretoriensis YK-1by column chromatography on Toyopearl HW55F, DEAE-Toyopearl 650M, hydroxylapatite and phenyl-Toyopearl 650M. Further purification by preparative polyacrylamide gel electrophoresis (PAGE)gave the homogenous protein, but the specific activity was reduced. The molecular weight of the enzyme was estimated to be 69,000 by SDS-PAGE and 60,000 by gel filtration. Optimum pH and temperature were 6.8 and 35℃, respectively. The enzyme was inhibited strongly by AgNO_3, HgCl_2, sodium dodecyl sulfate, and N-ethylmaleimide. The K_m (mM) for p-nitrophenyl α-D-glucopyranoside, maltose, maltotriose, isomaltose, methylα-glucoside, and sucrose were 0.15, 150, 45, 17, 18, and 29, and V_<max> (μmol / min / mg protein) for those substrates were 87, 0.23, 2.4, 9.0, 12, and 7.4, respectively. The N-terminal amino acid sequence of the enzyme was PEVKNHPETQPKWWKEATVY. The properties of α-glucosidase from T. pretoriensis YK-1 were similar to those from Saccharomyces cerevisiae.

• 社団法人日本農芸化学会の論文
• 1993-11-23

著者

• ODA Yuji

  Department of Food Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro

• Oda Y

  Fukuyama Univ. Hiroshima Jpn

• TONOMURA KENZO

  Department of Agricultural Chemistry, College of Agriculture, University of Osaka Prefecture

• HIROMI Keitaro

  Department of Food Science and Technology, Faculty of Engineering, Fukuyama University

• IWAMOTO Hiroyuki

  Department of Food Science and Technology, Faculty of Engineering, Fukuyama University

• Tonomura K

  Department Of Food Technology Faculty Of Engineering Fukuyama University

• Iwamoto Hisao

  Laboratory Of Food And Biomolecular Science Graduate School Of Agricultural Science Tohoku Universit

• Tonomura Kenzo

  Department Of Agricultural Chemistry College Of Agriculture University Of Osaka Prefecture

• Hiromi Keitaro

  Department Of Food Science And Technology Faculty Of Engineering Fukuyama University

• Hiromi Keitaro

  Department Of Food Science And Technology Faculty Of Agriculture Kyoto University

• Hirayama K

  Univ. Tokyo Tokyo Jpn

• Iwamoto H

  Laboratory Of Food And Biomolecular Science Graduate School Of Agricultural Science Tohoku Universit

• Oda Yuji

  Department Of Agricultural And Life Science Obihiro University Of Agriculture And Veterinary Medicin

• Iwamoto Hiroyuki

  Department Of Applied Biological Science Faculty Of Life Science And Biotechnology Fukuyama Universi

• HIROMI Keitaro

  Department of Food Science & Technology, Faculty of Agriculture, Kyoto University

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