Purification, Characterization, and Subsite Affinities of Thermoactinomyces vulgaris R-47 Maltooligosaccharide-metabolizing Enzyme Homologous to Glucoamylases

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概要

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• A maltooligosaccharide-metabolizing enzyme from Thermoactinomyces vulgaris R-47 (TGA) homologous to glucoamylases does not degrade starch efficiently unlike most glucoamylases such as fungal glucoamylases (Uotsu-Tomita et al., Appl. Microbiol. Biotechnol., 56, 465–473 (2001)). In this study, we purified and characterized TGA, and determined the subsite affinities of the enzyme. The optimal pH and temperature of the enzyme are 6.8 and 60°C, respectively. Activity assays with 0.4% substrate showed that TGA was most active against maltotriose, but did not prefer soluble starch. Kinetic analysis using maltooligosaccharides ranging from maltose to maltoheptaose revealed that TGA has high catalytic efficiency for maltotriose and maltose. Based on the kinetics, subsite affinities were determined. The A1+A2 value of this enzyme was highly positive whereas A4–A6 values were negative and little affinity was detected at subsites 3 and 7. Thus, the subsite structure of TGA is different from that of any other GA. The results indicate that TGA is a metabolizing enzyme specific for small maltooligosaccharides.

• 社団法人 日本農芸化学会の論文
• 2004-02-23

著者

• Aoki H

  Iwate Univ. Iwate Jpn

• Aoki Hideyuki

  Research Laboratory Ikeda Food Research Co. Ltd.

• NISHIKAWA ATSUSHI

  Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka U

• Aoki H

  Ikeda Food Res. Co. Ltd. Hiroshima Jpn

• TONOZUKA Takashi

  Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture an

• 殿塚 隆史

  東京農工大学農学部

• ICHIKAWA Kazuhiro

  Department of Applied Biological Science, Tokyo University of Agriculture Technology

• SAKANO Yoshiyuki

  Department of Applied Biological Science, Tokyo University of Agriculture Technology

• UOTSU (TOMITA)

  Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture an

• AKEBOSHI Hiromi

  Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture an

• Uotsu (tomita)

  Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And

• Tonozuka Takashi

  Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And

• Ichikawa Kimihisa

  Department Of Applied Microbiology Nippon Roche Research Center

• Nishikawa A

  Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And

• Nishikawa Atsushi

  Department Of Mechanical Science And Bioengineering Graduate School Of Engineering Science Osaka Uni

• Sakano Y

  Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And

• Sakano Yoshiyuki

  Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And

• Nishikawa Atsushi

  Department Of Applied Biological Science Tokyo University Of Agriculture And Technology

• Tonozuka Takashi

  Forage Crop Breeding Unit National Agricultural Research Center For Kyushu Okinawa Region

• Uotsu-Tomita Rie

  Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology

• Ichikawa Kazuhiro

  Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology

• SAKANO Yoshiyuki

  Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo Noko University

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