Thermoanaerobium brockiiが産生する新規ホスホリラーゼ,コージビオースホスホリラーゼの精製とその諸性質 Purification and Characterization of a Novel Phosphorylase, Kojibiose Phosphorylase, from Thermoanaerobium brockii

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Abstract

好熱嫌気性菌Thermoanaerobium brockii ATCC35047はコージビオースを可逆的に加リン酸分解し,β-グルコース1一リン酸とグルコースを生成する新規酵素,コージビオースホスホリラーゼを産生する.本酵素を各種カラムクロマトグラフィーにより,菌体破砕抽出液から電気泳動的に単一にまで精製した.本酵素の分子量は83,000,等電点は4.3-4.4であった.本酵素の至適pHは5.5,至適温度は65℃ であった.pH安定性は5.5-9.7,温度安定性は65℃ までであった.本酵素の活1生はHg<SUP>2+</SUP>イオンとPb<SUP>2+</SUP>イオンで阻害された.コージビオース,無機リン酸,グルコース,炉グルコース1-リン酸に対するK<SUP>m</SUP>値はそれぞれ0.77mM,0.85mM,3.52mM,0.77mMであった.

The thermophilic anaerobe Thermoanaerobium brockii ATCC 35047 produces a novel phosphorylase, kojibiose phosphorylase, which catalyzes the reversible phosphorolysis of kojibiose to form β-glucose 1-phosphate and D-glucose. The enzyme was purified from a cell-free extract to an electrophoretically homogeneous state by successive column chromatography on DEAE-Toyopearl 650S, CM-Toyopearl 6505, Hydroxyapatite, Ultrogel AcA44, Mono Q, and Butyl-Toyopearl 650 M . The enzyme had a molecular weight of 83, 000 by SDS-polyacrylamide gel electrophoresis and a pl of 4 .3 to 4.4 by gel isoelectrofocusing. The enzyme showed the highest activity at pH 5 .5 and 65°C, and was stable from pH 5.5 to 9.7 and up to 65°C. The enzyme activity was inhibited by Hg<SUP>2+</SUP> and Pb<SUP>2+0</SUP>. The Km values for kojibiose, Pi, glucose, and β-glucose 1-phosphate were 0 .77, 0.85, 3.52, and 0.77 mM, respectively.

Journal

  • Journal of Applied Glycoscience

    Journal of Applied Glycoscience 46(4), 423-429, 1999-12-01

    The Japanese Society of Applied Glycoscience

References:  15

Cited by:  9

Codes

  • NII Article ID (NAID)
    10008259401
  • NII NACSIS-CAT ID (NCID)
    AN10453916
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    13403494
  • NDL Article ID
    4976595
  • 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  CJPref  NDL  J-STAGE  JASI 
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