Fluorescent Activity Assay of the Enzymes Hydrolyzing Acidic Polysaccharides
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概要
- 論文の詳細を見る
Based on the amide linkage formation catalyzed by water-soluble carbodiimide, EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide), various acidic polysaccharides could be labeled by the fluorescent probe of EDAN (N-1-ethylenediaminonaphthalene). The pectinase reaction was evaluated by the EDC-OPA (o-phthalaldehyde) method [Anal. Biochem. , 189, 122-125 (1990)]. Moreover, paper chromatographic purification of the glucuronic acid-EDAN adduct allowed a 57-fold increase in the sensitivity of de-tection limit, i. e. , about 0. 19 nmol of glucuronate could be measured. Pectin-EDAN adduct worked as the substrate for pectinase and changes in the molecular weight distribution during the course of pectinase digestion were monitored by gel filtration with a Sepharose 6B column. In the case of hyaluronic acid-EDAN adduct, the fluorescent substrate was unfavorable to the hyaluronidase action and 2 to 5-fold higher K_ms were obtained with the hyaluronic acids of lower and higher contents of EDAN, respectively.
- 社団法人日本農芸化学会の論文
- 1994-02-23
著者
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KOBAYASHI Mikihiko
National Food Research Institute
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CHIBA Yogo
National Food Research Institute
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Chiba Y
Univ. Shizuoka Shizuoka Jpn
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