2012 Fiscal Year Final Research Report
Conversion of an inverting exo-D-glucosaminidase into a golycosynthase
| Project/Area Number |
22780095
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied biochemistry
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| Research Institution | Ishikawa Prefectural University |
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Principal Investigator |
HONDA Yuji 石川県立大学, 生物資源環境学部, 准教授 (40399382)
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| Project Period (FY) |
2010 – 2012
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| Keywords | エキソβ-D-グルコサミニダーゼ |
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| Research Abstract |
We elucidated an inverting exo-β-D-glucosaminidase activity of the GH9 protein from Photobacterium profundum SS9. Kinetic analysis of the hydrolytic activities toward cellooligosaccharides and chitooligosaccharides indicated the enzyme is an exo-β-D-glucosaminidase (EC 3.2.1.165) rather than an exo-β-1,4-glucosidase.Based on the modelled three-dimensional structure, we targeted D139, D143, and E555 as candidates of the important amino acid residues in the hydrolytic reaction. D139A, D143A and E555A mutants displayed trace hydrolytic activities. The D139A displayed glycosynthase activity with α-GlcN-F as a substrate.
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