Enzymatic Characteristics of Cellobiose Phosphorylase from <I>Ruminococcus albus</I> NE1 and Kinetic Mechanism of Unusual Substrate Inhibition in Reverse Phosphorolysis
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- HAMURA Ken
- Research Faculty of Agriculture, Hokkaido University
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- SABURI Wataru
- Research Faculty of Agriculture, Hokkaido University
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- ABE Shotaro
- Research Faculty of Agriculture, Hokkaido University
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- MORIMOTO Naoki
- Research Faculty of Agriculture, Hokkaido University
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- TAGUCHI Hidenori
- Research Faculty of Agriculture, Hokkaido University
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- MORI Haruhide
- Research Faculty of Agriculture, Hokkaido University
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- MATSUI Hirokazu
- Research Faculty of Agriculture, Hokkaido University
Bibliographic Information
- Other Title
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- Enzymatic Characteristics of Cellobiose Phosphorylase from Ruminococcus albus NE1 and Kinetic Mechanism of Unusual Substrate Inhibition in Reverse Phosphorolysis
- Enzymatic characteristics of cellobiose phosphorylase from Ruminococcus albus NE1 and kinetic mechanism of unusual substrate inhibition in reverse phophorolysis
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Abstract
Cellobiose phosphorylase (CBP) catalyzes the reversible phosphorolysis of cellobiose to produce α-D-glucopyranosyl phosphate (Glc1P) and D-glucose. It is an essential enzyme for the metabolism of cello-oligosaccharides in a ruminal bacterium, Ruminococcus albus. In this study, recombinant R. albus CBP (RaCBP) produced in Escherichia coli was characterized. It showed highest activity at pH 6.2 at 50 °C, and was stable in a pH range of 5.5–8.8 and at below 40 °C. It phosphorolyzed only cellobiose efficiently, and the reaction proceeded through a random-ordered bi bi mechanism, by which inorganic phosphate and cellobiose bind in random order and D-glucose is released before Glc1P. In the synthetic reaction, RaCBP showed highest activity to D-glucose, followed by 6-deoxy-D-glucose. D-Mannose, 2-deoxy-D-glucose, D-glucosamine, D-xylose, 1,5-anhydro-D-glucitol, and gentiobiose also served as acceptors, although the activities for them were much lower than for D-glucose. D-Glucose acted as a competitive-uncompetitive inhibitor of the reverse synthetic reaction, which bound not only the Glc1P site (competitive) but also the ternary enzyme-Glc1P-D-glucose complex (uncompetitive).
Journal
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 76 (4), 812-818, 2012
Japan Society for Bioscience, Biotechnology, and Agrochemistry
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Details 詳細情報について
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- CRID
- 1390001206478547968
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- NII Article ID
- 10030752061
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- NII Book ID
- AA10824164
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- COI
- 1:STN:280:DC%2BC38rjt1Wgsg%3D%3D
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- ISSN
- 13476947
- 09168451
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- NDL BIB ID
- 023593189
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- PubMed
- 22484959
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed