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KMID : 0903520000430020100
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
2000 Volume.43 No. 2 p.100 ~ p.105
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Glucose Determination by Using Korean Radish Anionic Peroxidase
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Abstract
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Anionic peroxidases (POD) were isolated from Korean radish (Raphanus sativus L.) root by using fractionation with (NH₄)₂SO₄ and CM-cellulose ion exchange chromatography and used as the colorimetric enzyme for glucose determination. The chromogen used in this work was o-tolidine or 4-aminoantipyrine/diethylaniline (4AA/DEA) and the colored products were measured at 630 ㎚. Korean radish anionic POD showed much better colorimetric reaction of glucose determination with 4AA/DEA than with o-tolidine. The r values of calibration curve for glucose determination by o-tolidine and 4AA/DEA were 0.9983 and 0.9963, respectively. In order to compare the reactivity for substrate oxidation by Korean radish POD and horseradish POD, the Km values against o-dianisidine and guaiacol were measured. Korean radish POD had about 40 fold higher affinity for o-dianisidine and 2 fold higher affinity for guaiacol as revealed by Km values. These results showed that Korean radish POD could be developed as the colorimetric diagnosis reagent for glucose determination with high sensitivity.
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