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Colorimetric Quantification of β-(1→4)-Mannobiose and 4-O-β-D-Mannosyl-D-glucose
Title: | Colorimetric Quantification of β-(1→4)-Mannobiose and 4-O-β-D-Mannosyl-D-glucose |
Authors: | Jaito, Nongluck Browse this author | Saburi, Wataru Browse this author →KAKEN DB | Muto, Hirohiko Browse this author | Matsui, Hirokazu Browse this author →KAKEN DB | Mori, Haruhide Browse this author →KAKEN DB |
Keywords: | oligosaccharide quantification | β-(1→4)-mannobiose | 4-O-β-D-mannosyl-D-glucose | cellobiose 2-epimerase | 4-O-β-D-mannosyl-D-glucose phosphorylase |
Issue Date: | 20-Nov-2014 |
Publisher: | 日本応用糖質科学会 |
Journal Title: | Journal of Applied Glycoscience |
Volume: | 61 |
Issue: | 4 |
Start Page: | 117 |
End Page: | 119 |
Publisher DOI: | 10.5458/jag.jag.JAG-2014_007 |
Abstract: | Spectrophotometric quantification method of carbohydrates is useful for processing multiple samples. In this study, we established colorimetric quantification for 4-O-β-D-mannosyl-D-glucose (Man-Glc) and β-(1→4)-mannobiose (Man2). For quantification of Man-Glc, phosphorolysis of Man-Glc catalyzed by 4-O-β-D-mannosyl-D-glucose phosphorylase (MGP) was coupled with quantification of D-glucose by the glucose oxidase-peroxidase method. In addition to MGP, cellobiose 2-epimerase (CE) was added for quantification of Man2. In both quantifications, a good linear relationship was obtained between A505 and the sample concentration (0-0.5 mM). The A505 values obtained at various concentrations of Man2 and Man-Glc were almost identical to those with equivalent D-glucose concentrations. Kinetic parameters of Ruminococcus albus and Rhodothermus marinus CEs for the epimerization of Man2 were determined using the quantification method for Man-Glc. Both enzymes showed 5-15-fold higher kcat/Km values than those for cellobiose and lactose, which supports the prediction that these enzymes utilize Man2 as a substrate in the β-mannan metabolic pathway. |
Type: | article (author version) |
URI: | http://hdl.handle.net/2115/67698 |
Appears in Collections: | 農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 佐分利 亘
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