Characteristics of C. vulgaris CE and MGP
9
stopped by heating the sample at 100 °C for 3 min, and
the reaction mixture was desalted with Amberlite MB-4
[9] Ito S, Hamada S, Yamaguchi K, Umene S, Ito H, Matsui H,
Ozawa T, Taguchi H, Watanabe J, Wasaki J, Ito S. Cloning and
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merase from a thermohalophilic bacterium, Rhodothermus mari-
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(
Roam and Haas, Philadelphia, PA, USA). Resulting sam-
ple was used as the substrate of CvMGP.
The phosphorolytic activity of CvMGP to cellobiose
and lactose was measured based on the amount of
D-glucose liberated. A reaction mixture (50 μL) con-
taining 151 nM CvMGP, 2 mM cellobiose or lactose,
[
[
[
1
0
00 mM sodium phosphate buffer (pH 8.0), and
.2 mg/mL BSA was incubated at 37 °C for 10 min.
D-Glucose liberated was measured as described above.
Acknowledgments
Part of this work was supported by JSPS KAKENHI
with a Grants-in-Aid for Young Scientists (B) (grant
number 26850059). We would like to thank Tomohiro
Hirose of the Instrumental Analysis Division, Equip-
ment Management Center, Creative Research Institute,
Hokkaido University, and the staff of the DNA
Sequencing Facility of the Research Faculty of Agricul-
ture, Hokkaido University for amino acid and DNA
sequence analyses, respectively.
[13] Park CS, Kim JE, Choi JG, Oh DK. Characterization of a
recombinant cellobiose 2-epimerase from Caldicellulosiruptor
saccharolyticus and its application in the production of mannose
from glucose. Appl. Microbiol. Biotechnol. 2011;92:1187–1196.
[
14] Kim JE, Kim YS, Kang LW, Oh DK. Characterization of a
recombinant cellobiose 2-epimerase from Dictyoglomus turgidum
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charides. Biotechnol. Lett. 2012;34:2061–2068.
[15] Ojima T, Saburi W, Yamamoto T, Mori H, Matsui H. Identifica-
tion and characterization of cellobiose 2-epimerases from various
aerobes. Biosci. Biotechnol. Biochem. 2013;77:189–193.
[
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16] Park CS, Kim JE, Lee SH, Kim YS, Kang LW, Oh DK. Charac-
Funding
terization of a recombinant mannobiose 2-epimerase from Spiro-
chaeta thermophila that is suggested to be
-epimerase. Biotechnol. Lett. 2013;35:1873–1880.
a cellobiose
Part of this work was supported by JSPS KAKENHI with
a Grants-in-Aid for Young Scientists (B) [grant number
2
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