2001 Fiscal Year Final Research Report Summary
Development of New Chemo-enzymatic Synthetic Process of Glycoconjugates
| Project/Area Number |
11650825
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
生物・生体工学
|
|---|
| Research Institution | The Noguchi Institute |
|---|
Principal Investigator |
HANEDA Katsuji The Noguchi Institute, Senior Researcher, 研究部, 主任研究員 (40270540)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
野口 緑(竹内緑) (財)野口研究所, 研究部, 研究員
NOGUCHI Midori The Noguchi Institute, Researcher
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Keywords | Glycoconjugate / Oligosaccharide / Chemo-enzymatic / Peptide / Endoglycosidase / Transglycosylation / Endo-M / Immobilized enzyme |
|---|
| Research Abstract |
The new chemo-enzymatic synthetic method of glycoconjugates was developed. This method is based on chemical synthesis of acceptor substrate containing a glycoside (GlcNAc) moiety and the transglycosylation of olligosaccharide block to it catalyzed by endoglycosidase (Scheme 1).
R-GlcNAc-GlcNAc-Asu + GlcNAc-X → R-GlcNAc-GlcNAc-X + GlcNAc-Asn (Scheme 1)
[Glycoside donor] [Acceptor] Endo-M[Transglycosylation product] R: Oligosaccaride, X: Peptide etc.
In this research, we tried to develop this technique as the practical synthetic method of neo-glycoconjugates based on the following two experiments :
1. Synthesis of various model glycoconjugates
1st step : Synthesis of various model glycoconjugates
2nd step : Establishment of a practical synthetic method : Solid-phase method
1) Using endo-β-N-acetylglucosaminidase of Mucor hiemalis (Endo-M), the transglycosylation product could be obtained in a high yield.
2) The introduction of oligosaccharide not only into asparagine (Asn) residue but also into glutamine (Gln) residue of a peptide, to which biological addition of oligosaccharide is impossible, became possible. Plural same or different kinds of oligosaccharides could be efficiently introduced into the peptide as a model of glycoprotein.
2. Establishment of practical synthetic method : Solid-phase method
1) Among the two ideas, which were based on the immobilization of the enzyme and on the immobilization of acceptor substrate, the process using the immobilized enzyme was studied.
2) Endo-M was immobilized on the resin, and the transglycosylation reaction using this enzyme was done.
3) The model recycle process based on the transglycosylation reaction by immobilized enzyme, the isolation of the objective reaction product, and the reuse of recovered residual substrates was proposed.
|
