Identification of Catalytic Amino Acids of Cyclodextran Glucanotransferase from<i>Bacillus circulans</i>T-3040

• YAMAMOTO Tomoko

  National Food Research Institute

• TERASAWA Kazue

  National Food Research Institute

• KIM Young-Min

  Laboratory of Molecular Enzymology, Hokkaido University

• KIMURA Atsuo

  Laboratory of Molecular Enzymology, Hokkaido University

• KITAMURA Yoshiaki

  National Food Research Institute

• KOBAYASHI Mikihiko

  Jissen Women’s University

• FUNANE Kazumi

  National Food Research Institute

書誌事項

タイトル別名

• Identification of Catalytic Amino Acids of Cyclodextran Glucanotransferase from Bacillus circulans T-3040

抄録

In glycoside hydrolase family 66 (see http://afmb.cnrs-mrs.fr/CAZY/), cyclodextran glucanotransferase (CITase) is the only transglycosylation enzyme, all the other family 66 enzymes being dextranases. To analyze the catalytic amino acids of CITase, we modified CITase chemically from the T-3040 strain of Bacillus circulans with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). EDC inactivated the enzyme by following pseudo-first order kinetics. In addition, the substrates of an isomaltooligosaccharide and a cyclodextran inhibited EDC-induced enzyme inactivation, implicating the carboxyl groups of CITase as the catalytic amino acids of the enzyme. When two conserved aspartic acid residues, Asp145 and Asp270, were replaced with Asn in T-3040 mature CITase, CIT-D270N was completely inactive, and CIT-D145N had reduced activity. The V_max of CIT-D145N was 1% of that of wild-type CITase, whereas the K_m of CIT-D145N was about the same as that of the wild-type enzyme. These findings indicate that Asp145 and Asp270 play an important role in the enzymatic reaction of T-3040 CITase.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 70 (8), 1947-1953, 2006

  公益社団法人 日本農芸化学会

キーワード

• cyclodextran
• cyclodextran glucanotransferase
• catalytic amino acid
• aspartate residue

詳細情報

• CRID: 1390001206479478144
• NII論文ID: 10018528285
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.60105
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 8040467
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I8040467
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.60105
• 本文言語コード: en
• データソース種別:
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