Cloning, Sequencing, and Expression of the Genes Encoding an Isocyclomaltooligosaccharide Glucanotransferase and an α-Amylase from a Bacillus circulans Strain

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The gene for a novel glucanotransferase, isocyclomaltooligosaccharide glucanotransferase (IgtY), involved in the synthesis of a cyclomaltopentaose cyclized by an α-1,6-linkage [ICG5; <I>cyclo</I>-{→6)-α-<small>D</small>-Glc<I>p</I>-(1→4)-α-<small>D</small>-Glc<I>p</I>-(1→4)-α-<small>D</small>-Glc<I>p</I>-(1→4)-α-<small>D</small>-Glc<I>p</I>-(1→4)-α-<small>D</small>-Glc<I>p</I>-(1→}] from starch, was cloned from the genome of <I>B. circulans</I> AM7. The IgtY gene, designated <I>igtY</I>, consisted of 2,985 bp encoding a signal peptide of 35 amino acids and a mature protein of 960 amino acids with a calculated molecular mass of 102,071 Da. The deduced amino-acid sequence showed similarities to 6-α-maltosyltransferase, α-amylase, and cyclomaltodextrin glucanotransferase. The four conserved regions common in the α-amylase family enzymes were also found in this enzyme, indicating that this enzyme should be assigned to this family. The DNA sequence of 8,325-bp analyzed in this study contained two open reading frames (ORFs) downstream of <I>igtY</I>. The first ORF, designated <I>igtZ</I>, formed a gene cluster, <I>igtYZ</I>. The amino-acid sequence deduced from <I>igtZ</I> exhibited no similarity to any proteins with known or unknown functions. <I>IgtZ</I> was expressed in <I>Escherichia coli</I>, and the enzyme was purified. The enzyme acted on maltooligosaccharides that have a degree of polymerization (DP) of 4 or more, amylose, and soluble starch to produce glucose and maltooligosaccharides up to DP5 by a hydrolysis reaction. The enzyme (IgtZ), which has a novel amino-acid sequence, should be assigned to α-amylase. It is notable that both IgtY and IgtZ have a tandem sequence similar to a carbohydrate-binding module belonging to a family 25. These two enzymes jointly acted on raw starch, and efficiently generated ICG5.

収録刊行物

  • Bioscience, biotechnology, and biochemistry  

    Bioscience, biotechnology, and biochemistry 70(11), 2690-2702, 2006-11-23 

    Japan Society for Bioscience, Biotechnology, and Agrochemistry

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各種コード

  • NII論文ID(NAID)
    10018524871
  • NII書誌ID(NCID)
    AA10824164
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    09168451
  • NDL 記事登録ID
    8560481
  • NDL 雑誌分類
    ZR7(科学技術--農林水産--農産) // ZR2(科学技術--生物学--生化学) // ZP1(科学技術--化学・化学工業)
  • NDL 請求記号
    Z53-G223
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
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