Acetobacter capsulatum ATCC 11894株起源の菌体外デキストリンデキストラナーゼの簡易精製法と基本性質 Simple Purification and Characterization of an Extracellular Dextrin Dextranase from Acetobacter capsulatum ATCC 11894

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Abstract

酢酸菌Acetobacter capsulaturn ATCC11894株は,グルコースと微量のデキストリンを炭素源として液体培養すると,菌体外にグルコシル基転移酵素デキストリンデキストラナーゼ(EC2.4.1.2)を産生することが知られている.当該酵素は,各種クロマトグラフィーの操作を経ることなく,高速冷却遠心分離(4℃,20,000×9,20分間)のみにて,簡易かつ高純度に精製された.精製酵素の分子量はSDS-PAGE法から,約152kDaと推定された.また,当該酵素は,pH5.2および38℃ において最大活性を示した.精製酵素を各種pHで4℃,24時間処理すると,pH4.1-5.4の範囲で100%の残存活性が認められた.本酵素活性は,1mMのHg2+,Pb2+あるいはKMnO4により完全に阻害された.当該酵素により,マルース以外の一連のマルトオリゴ糖から生成されるα-1,6-グルカンは,1H-NMR分析の結果から,わずかにα-1,4-グルコシド結合の分枝を有することが示唆された.生成α-1,6-グルカンの分子量は,およそ1270kDaと推定された.

Dextrin dextranase (DDase, EC2.4.1.2) was extracellularly secreted in a culture medium of Acetobacter capsulatum ATCC 11894 containing both glucose (5.45%) and an extremely small amount of dextrin (0.05%) as the essential carbon sources. The enzyme was simply purified by only one-step centrifugation at 4°C and 20, 000×g for 20 min, and immediately dialyzed against 50 mM acetate buffer (pH 4.5) at CC for 2-3 days. The purified DDase gave a single protein band on both Native- and SDS-PAGE. The molecular mass of the enzyme was estimated to be about 152 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme were 5.2 and 38°C, respectively. The enzyme retained its original activity up to 45°C, and was stable in the range of pH 4.1-5.4 at 4°C for 24 h. The enzyme was completely inactivated by 1 mM of Hg2+ Pb2+ or KMn04. The purified enzyme effectively synthesized α-1, 6-glucan from maltooligosaccharides. The average molecular mass of the product dextran was estimated to be about 1270 kDa.

Journal

  • Journal of Applied Glycoscience

    Journal of Applied Glycoscience 46(4), 469-473, 1999-12-01

    The Japanese Society of Applied Glycoscience

References:  16

Codes

  • NII Article ID (NAID)
    10008259525
  • NII NACSIS-CAT ID (NCID)
    AN10453916
  • Text Lang
    ENG
  • Article Type
    NOT
  • ISSN
    13403494
  • NDL Article ID
    4976745
  • NDL Source Classification
    ZP24(科学技術--化学・化学工業--糖・澱粉)
  • NDL Call No.
    Z17-15
  • Data Source
    CJP  NDL  J-STAGE  JASI 
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