Branched functional β-glucan synthesis using the glycosyltransferase activity of mushrooms glucanase View Homepage


Ontology type: schema:MonetaryGrant     


Grant Info

YEARS

2011-2013

FUNDING AMOUNT

4550000 JPY

ABSTRACT

While some β-glucan contained in mushrooms exhibit an anti-tumor effect, purification difficulties, the structure is complicated, because there is no means for synthesizing model material, it has not progressed elucidation of the molecular level about the effects. In this study, using the glycosyltransferase activity of the β-glucanase isolated from mushrooms, it was intended to establish a method to artificially design a variety of β-glucan. Heisei 24 fiscal cloning a new basidiomycete Schizophyllum commune from a carbohydrate hydrolase (GH) belonging to the family 30 β-1,6-glucanase (ScPus30A), was heterologous expression in Koji fungus (Aspergillus oryzae). Molecular weight on SDS-PAGE was estimated as 57,000. From the multi-alignment with other GH30 enzymes, 207 th and 302 th glutamic acid is predicted to be active center residue. Therefore, alanine, glycine, creating variants of serine heterologously expressed for each. Since it was examined synthesis of beta-1, 6-glucan with these mutant enzymes and fluoride sugar (.alpha. fluoride glucopyranoside), the synthesis reaction was not observed, the synthesis reaction using the present enzyme group It was expected to require fluoride gentiooligosaccharide. So also for the GH family two kinds of rice blast fungus was obtained a total of six types of mutant enzyme performs a synthetic enzyme into for the synthesis of oligosaccharides. Enzymatic synthesis of β-glucan by combining these are considered to be possible. On the other hand, the enzyme belonging to the Basidiomycetes cormorant signaling Seo Hee Toyota Ke than GH family 16 were cloned, was heterologous expression in bacteria Koji in the same manner as described above. Laminarin is a β-1,3 / 1,6-glucan derived from kelp was not degraded by this enzyme, β-1,3-glucan 3 sugar and has a glucose side chain in the β-1,6 bond as the main product 4 could be obtained sugar. There is a possibility that can be prepared the functionality glucan these oligosaccharides in the material. More... »

URL

https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23780169

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