1999 Fiscal Year Final Research Report Summary
Construction of a soluble "glycochain-recognition protein" structure and regulation of embrionic/cellular mechanism using the protein apparatus
| Project/Area Number |
09660359
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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 molecular and cellular biology
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| Research Institution | Prefecture University of Kyoto |
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Principal Investigator |
OHNISHI Masatake Prefecture University of Kyoto, Department of Biological Resources Chemistry, Professor, 農学部, 教授 (90026576)
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| Project Period (FY) |
1997 – 1999
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| Keywords | β-galactosidase / β-glucocerebrosidase / Yeast / Japanese cattle / taste sense receptor / sulfite dehydrogenase / Thiobacillus thiooxidans / β-1, 3-exo-glucanase |
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| Research Abstract |
(1) β-Glycosidases from yeast (Kluyvelomyces lactis, Saccharomyces cerevisiae) : β-Galactosidases(GAL) and β-1, 3-exo-glucanase(LAM) were purified to be a single protein from K. lactis and S. cerevisiae, respectively, and were found to strongly catalyze their trasglycosilation reactions as follows ; GAL produces alkyl galactosides with alchols and galactosyl glyceride with glycerol. LAM produces laminari-oligosaccharides with laminari-biose and -triose. (2) β-Glucocerebrosidase(GCase) from cattle brain : GCase was purified in a soluble state(S) to analyze its enzymatic properties, and was immobilized on a carrier (decyl sepharose) to make a "glycochain-rcognition" structure. (3) Taste sense receptor protein from cattle tongue : Two receptor proteins, MW 58,000 and 64,000 were purified and treated with protease to have peptide fractions. Two major peptides each were obtained and analyzed their sequences to take a genetic operation for large scale production of these proteins. (4) Fish (Oryzia Latipes) germ cells : Freezing of sperm cells and egg cell culture were succeeded for around 4 days. Mechanism on reception of the fertilization signal (sperm cells) to the receptor on egg cells was investigated using the biological microscope obtained in this research project. (5) Sulfite dehydrogenase from Thibacillus thiooxidans : The microbe was employed for making of a high sensitive and automatical sulfite-analyzer as a sensor. Thus enzymatic bases (sulfite dehydrogenase-catalyzed reaction) were nedded to investigate about the microbial state using the kinetic procedures. Based on the subcellular fractionation, the enzyme was confirmed to be a membrane-bound state of organelles. It is very possible to constrast a "membrane-bound structure" for the analytical apparatus.
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