Visual Assay for Specific Inhibitors of Adenosine Deaminase with Agar Plates Containing pH Indicator
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概要
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We have devised a simple, rapid method for detection and determination of specific inhibitors of adenosine deaminase, which involves paper disks of the inhibitors and agar plates containing adenosine, adenosine deaminase, and a pH indicator. The time required for a large-scale determination (e.g., a few dozen samples/9-cm Petri dish) is short (4 h or so). We first developed a paper disk-agar plate method for detecting adenosine deaminase activity using agar plates containing adenosine and a pH indicator. This was adapted from the method for zymoautography of this enzyme in electrophoresis gels and its modification with paper disks and agar gels contained in a multi-well microculture tray. Application was made of the paper disk - agar plate method following the models of paper disk - agar plate antibiotic bioassay and agar plate bioautography, to provide a method to detect and roughly determine the specific inhibitors of adenosine deaminase in large-scale work, and a method to locate the inhibitor activity in thin-layer chromatograms, respectively. This method is advantageous when used as a tool to detect activity in a large number of column chromatography fractions. After a few hours of practice, the inhibitor can be detected at a glance. Use here of this method is illustrated with J-518S substance, which is probably a new specific inhibitor of adenosine deaminase. The application of this method to detection of cytosine deaminase activity is also discussed.
- 社団法人日本生物工学会の論文
- 1985-10-25
著者
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KATSURAGI Tohoru
Graduate School of Biological Sciences, Nara Institute of Science and Technology
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Sakai Takuo
Laboratory Of Food Microbiological Engineering Faculty Of Agriculture Kinki University
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Sakai Takuo
Fermentation Chemistry Laboratory College Of Agriculture University Of Osaka Prefecture
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Sakai Takuo
Center For Research And Development Of Bioresources University Of Osaka Prefecture
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Sakai Takuo
Department Of Food Science Faculty Of Agriculture Kinki University
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Sakai Takuo
Department Of Applied Biological Chemistry College Of Agriculture University Of Osaka Prefecture
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Sakai Takuo
Department Of Applied Biochemistry Faculty Of Agriculture Osaka Prefecture Univeisity
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Katsuragi Tohoru
Fermentation Chemistry Laboratory College Of Agriculture University Of Osaka Prefecture
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Katsuragi Tohoru
Graduate School Of Biological Sciences Nara Institute Of Science And Technology (naist)
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KATSURAGI TOHORU
The University of Osaka Prefecture, College of Agriculture, Laboratory of Fermentation Chemistry
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SAKAI TAKUO
The University of Osaka Prefecture, College of Agriculture, Laboratory of Fermentation Chemistry
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TONOMURA KENZO
The University of Osaka Prefecture, College of Agriculture, Laboratory of Fermentation Chemistry
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Tonomura K
Department Of Food Technology Faculty Of Engineering Fukuyama University
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