Fluorometric quantification of total d-gluconate by a flow-injection system using an immobilized-enzyme reactor |
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Authors: | Tadayuki Tsukatani Kiyoshi Matsumoto |
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Affiliation: | aBiotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Kurume 839-0861, Japan;bDepartment of Bioscience and Biotechnology, Division of Food Biotechnology, Faculty of Agriculture, Graduate School Kyushu University, Fukuoka 812-8581, Japan |
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Abstract: | A method for the quantification of total d-gluconate by flow-injection analysis was developed using an immobilized-enzyme reactor and fluorescence detection. d-Gluconate was quantified using co-immobilized gluconate kinase (GK) and 6-phosphogluconate dehydrogenase (PGDH) reactor. d-Gluconate was phosphorylated to 6-phospho-d-gluconate by GK in the presence of ATP, and then the 6-phospho-d-gluconate produced was oxidized by PGDH with NADP+. The NADPH produced by the GK-PGDH reactor was monitored fluorometrically at 455 nm (excitation at 340 nm). A linear relationship between the responses and concentrations of d-gluconate was obtained in the ranges of 1.0 × 10−6–1.6 × 10−4 M. The relative standard deviation for 10 successive injections was 0.57% at the 0.1 mM level. This analytical method was applied to the quantification of d-gluconate in honeys, vinegars and noble rot wines, and the results showed good agreement with those obtained using the conventional F-kit method. |
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Keywords: | Flow-injection analysis Immobilized-enzyme reactor smCaps" >d-Gluconate Honey Vinegar Noble rot wine |
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