アミラーゼおよびプロテアーゼ活性の半定量的簡易検出法の開発と応用  [in Japanese] Semi-quantitative assay of amylase and protease activities using agar plates with enzyme substrates.  [in Japanese]

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Abstract

<p>高校生物のカリキュラムでは,酵素の働きや生物学的な役割について取り扱っている.しかし,基本的な酵素特性を調べる教材は乏しい.酵素をより良く理解させるためには,簡単,且つ,定量的に測定できる酵素検出法の開発が望まれる.可溶性デンプンやカゼインを含む寒天プレートを用いることにより,アミラーゼおよびプロテアーゼ活性を検出することができた.アミラーゼ活性を調べるために,Aspergillus oryzaeからの酵素標品,発芽中のイネ・ハツカダイコン種子,胃腸薬,ヒト唾液を用いた.プロテアーゼについては, A. melleusからの酵素標品,衣料用洗剤,パイナップル・キウイフルーツの果肉,キノコ子実体を用いた.酵素反応後,寒天プレートをヨード液に浸漬することにより,アミラーゼ活性を検出した.一方,プロテアーゼ活性の検出には,寒天プレートを混合有機溶剤(メタノール:酢酸:水=40:10:50)に浸漬した.その結果,両酵素ともそれらの活性を半定量的に可視化することができた.簡単な実験操作であり,実験経費も限られており,特別な機器類も要しないことから,この酵素検出法は教育現場で使用できる有効な方法であると帰結した.</p>

<p>The functional concept and biological properties of enzymes are included in the biology curriculum of high school. However, there have been few educational materials available on fundamental enzymology. Easy and quantitative assay systems of enzyme activities are required to lead students to better understanding of enzymes. Amylase and protease activities were assayed using the agar plates with soluble starch and casein. An enzyme preparation from Aspergillus oryzae was tested for amylase assay as well as extracts from germinating seeds of rice and radish, a stomachic medicine, and human saliva. The protease assay was carried out using an enzyme preparation from A. melleus and various extracts from a laundry detergent, fruit meats of pineapple and kiwifruit, and a mushroom fruit body. After enzyme reaction, the agar plates were immersed in the iodine solution for amylase detection, and in the mixed solvents of methanol : acetic acid : water= 4:1:5 for protease detection. Obtained results showed that semi-quantitative activities of both enzymes were visualized by the immersion treatment, concluding that this system is a useful tool due to easy handling, cheaper material costs and no special bioinstruments needed.</p>

Journal

  • Japanese Journal of Biological Education

    Japanese Journal of Biological Education 54(2), 60-67, 2014

    The Society of Biological Science Education of Japan

Codes

  • NII Article ID (NAID)
    130007719613
  • NII NACSIS-CAT ID (NCID)
    AN0012957X
  • Text Lang
    JPN
  • ISSN
    0287-119X
  • NDL Article ID
    025571812
  • NDL Call No.
    Z18-53
  • Data Source
    NDL  J-STAGE 
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