Fluorospectral Intensity of High-molecular-weight Dextrans: Application to the Enzyme Reactions
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概要
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A rapid and simple assay procedure of high-molecular-weight polysaccharides is required for the activity measurement of polysaccharide-synthesizing or -hydrolyzing enzymes. When an aqueous solution of dextran was assayed fluorospectrometrically at a fixed wavelength of 340 nm for both excitation and emission, a strong response based on Rayleighs light scattering was observed. There was a linear correlation between the spectral intensity and concentration of the native dextran, and the intensity was proportional to the molecular weight of the dextran. Two model experiments verified the applicability of fluorospectrometric analysis. That is, a kinetic measurement of dextran synthesis by this method corresponded well with the results evaluated by the reducing end group analysis. Differences between endo-type and exo-type dextranases were clearly demonstrated. Moreover, the endodextranase action on the dextran could be followed by the changes in spectral intensity of the incubation mixture placed in the cuvette. Although fluorospectrometric analysis is applicable to polysaccharides other than dextran, its use is restricted to homogeneous substrates or enzymes. High-performance liquid chromatography with a gelfiltration column indicated that the spectral intensity was specific for the high-molecular-weight dextran, while a differential refractometer gave responses to all sugars including those with low-molecular-weights.
- 社団法人 日本農芸化学会の論文
著者
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Kobayashi Mikihiko
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Matsuda Kazuo
Department Of Agricultural Biochemistry Faculty Of Agriculture Tohoku University
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UTSUGI Hideki
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
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