1998 Fiscal Year Final Research Report Summary
Screening of glycosidase that cleave mammalian glycolipid mimicking lipopolysacchrides
| Project/Area Number |
09660298
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | Tottori University |
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Principal Investigator |
NODA Ken Tottori University, Department of Biochemistry and Biotechnology, Associate Professor, 農学部, 助教授 (60218287)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASAKI Ryohei Tottori University, Department of Biochemistry and Biotechnology, Full Professor, 農学部, 教授 (80273887)
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| Project Period (FY) |
1997 – 1998
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| Keywords | lipopolysacchrides / glycolipid / liver / digestion / metabolism / detoxification |
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| Research Abstract |
In this research project we have developed a method for the analysis of very low molecular weight lipopolysaccharide molecule derived from Gram-negative bacteria by using linear gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We also succeeded to modify thiobarbituric acid assay method to be applicable for micro scale analysis of acidic sugar molecule.
By using above techniques, we had screened for glycosidase activity that cleave R-mutant lipopolysaccharides (R-LPS) that consist common core structure of lipooligosaccharide or lipopolysaccharide. When bovine liver cytosol fraction is incubated with R-LPS, we found that a carbohydrate molecule that is cleaved out from the LPS. A chemical analysis showed that the molecule has a phosphate group in it. The substrate specificity of this unidentified glycosidase will be clarified by further analysis of the carbohydrate molecule. By using linear gradient SDS-PAGE, we also found LPS molecule disappears in time dependent manner from the incubation mixture with bovine liver cytosol fraction. This phenomenon suggest the presence of unknown LPS metabolism or detoxification system in liver tissue.
These results could lead the researches on a detoxification system for LPS that is not yet well understood at present. The results may also contribute to understand physiological aspects of ruminant digestive system that contain huge amount of microorganism including Gram-negative bacteria that produce variety of toxic lipopolysaccharide molecules.
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