Purification, Characterization, and Sequence Analysis of Two α-Amylase Isoforms from Azuki Bean, Vigna angularis, Showing Different Affinity towards β-Cyclodextrin Sepharose

• MAR San San

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• MORI Haruhide

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• LEE Jin-Ha

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• FUKUDA Kenji

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• SABURI Wataru

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• FUKUHARA Arinobu

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• OKUYAMA Masayuki

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• CHIBA Seiya

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

• KIMURA Atsuo

  <i>Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University</i>

書誌事項

タイトル別名

• Purification, Characterization, and Sequence Analysis of Two .ALPHA.-Amylase Isoforms from Azuki Bean, Vigna angularis, Showing Different Affinity towards .BETA.-Cyclodextrin Sepharose
• Purification Characterization and Sequence Analysis of Two アルファ Amylase Isoforms from Azuki Bean Vigna angularis Showing Different Affinity towards ベータ Cyclodextrin Sepharose
• Purification, Characterization, and Sequence Analysis of Two α-Amylase Isoforms from Azuki Bean,<i>Vigna angularis</i>, Showing Different Affinity towards β-Cyclodextrin Sepharose

抄録

  Two α-amylase isoforms designated VAAmy1 and VAAmy2 were purified from cotyledons of germinating seedlings of azuki bean (Vigna angularis). VAAmy1 apparently had lower affinity towards a β-cyclodextrin Sepharose column than VAAmy2. Molecular weights of VAAmy1 and VAAmy2 were estimated to be 47,000 and 44,000, respectively. However, no considerable difference was found between them in effects of pH, temperature, CaCl₂, and EDTA, as well as the kinetic parameters for amylose (average degree of polymerization 17): k_cat, 71.8 and 55.5 s⁻¹, K_m, 0.113 and 0.097 mg/ml; for blocked 4-nitrophenyl α-D-maltoheptaoside: k_cat, 62.4 and 85.3 s⁻¹, K_m, 0.22 and 0.37 mM, respectively. Primary structures of the two enzymes were analyzed by N-terminal sequencing, cDNA cloning, and MALDI-TOF mass spectrometry, implying that the two enzymes have the same peptide. The results indicated that the low affinity of VAAmy1 towards β-cyclodextrin Sepharose was due to some modification on/near carbohydrate binding site in the limited sequence regions, resulting in higher molecular weight.<br>

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 67 (5), 1080-1093, 2003

  公益社団法人 日本農芸化学会

キーワード

• azuki bean α-amylase
• isoforms
• possible posttranslational modification
• carbohydrate binding site
• alternative splicing

詳細情報

• CRID: 1390282681452480512
• NII論文ID: 110002694182
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.67.1080
• COI: 1:CAS:528:DC%2BD3sXktlygtbc%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 6545858
• PubMed: 12834286
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I6545858
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.67.1080
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可