2013 Fiscal Year Final Research Report
Structural analysis of uncharacterized enzymes acting on glycans and its application to the production of oligosaccharides
| Project/Area Number |
23570132
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
TONOZUKA Takashi 東京農工大学, (連合)農学研究科(研究院), 准教授 (50285194)
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| Co-Investigator(Renkei-kenkyūsha) |
NISHIKWA Atsushi 東京農工大学, 大学院農学研究院, 教授 (30218127)
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| Project Period (FY) |
2011 – 2013
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| Keywords | 酵素 / 糖 / 糖質加水分解酵素 / グリコシンターゼ / 糖タンパク質糖鎖 / X線結晶構造解析 |
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| Research Abstract |
This study focuses on bacterial enzymes homologous to processing alpha-glucosidases I and II, both of which hydrolyze eukaryotic glycans. An enzyme from E. coli, YgjK, was successfully converted into a glycosynthase, and the enzyme efficiently produced Glc-alpha-1,2-Gal. The result suggests that the natural substrates of YgjK are likely to be sugars containing the Glc-alpha-1,2-Gal structure. We also studied related enzymes from Thermus and other bacterial species. This study shows that a combination of protein crystallography and glycosynthase reaction allows us to determine the natural substrates of carbohydrate-active enzymes, even if the substrates are not commercially available.
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[Journal Article] Structure of a bacterial glycoside hydrolase family 63 enzyme in complex with its glycosynthase product, and insights into the substrate specificity2013
Author(s)
Miyazaki, T., Ichikawa, M., Yokoi, G., Kitaoka, M., Mori, H., Kitano, Y., Nishikawa, A. and Tonozuka, T.
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Journal Title
FEBS J.
Volume: 280
Pages: 4560-4571
DOI
Peer Reviewed
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[Journal Article] Heterologous expression and characterization of processing α-glucosidase I from Aspergillus brasiliensis ATCC 96422011
Author(s)
Miyazaki, T., Matsumoto, Y., Matsuda, K., Kurakata, Y., Matsuo, I., Ito, Y., Nishikawa, A. and Tonozuka, T.
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Journal Title
Glycoconj. J.
Volume: 28
Pages: 563-571
DOI
Peer Reviewed
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[Presentation] Conformational changes and substrate recognition of an uncharacterized glycosidase YgjK belonging to GH63 from Escherichia coli2014
Author(s)
Takatsugu Miyazaki, Mugumi Ichikawa, Haruhide Mori, Motomitsu Kitaoka, Yoshikazu Kitano, Atsushi Nishikawa, and Takashi Tonozuka Miyazaki, T., Ichikawa, M., Mori, H., Kitaoka, M., Kitano, Y., Nishikawa, A. and Tonozuka, T.
Organizer
10th Carbohydrate Bioengineering Meeting
Place of Presentation
Prague, Czech Republic
Year and Date
20140421-24
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