キムラ　マサヒロ   木村　将大    所属   応用生物学部 応用生物学科    職種   助教
言語種別	英語
発行・発表の年月	2017/05
形態種別	学術論文
査読	査読あり
標題	Improved fluorescent labeling of chitin oligomers: Chitinolytic properties of acidic mammalian chitinase under somatic tissue pH Conditions.
執筆形態	共著
掲載誌名	Carbohydr Polym
掲載区分	国外
出版社・発行元	Elsevier社
巻・号・頁	164,pp.145-153
著者・共著者	Wakita, S., Kimura, M. (equal contribution), Kato, N., Kashimura, A., Kobayashi, S., Kanayama, N., Ohno, M., Honda, S., Sakaguchi, M,. Sugahara, Y., Bauer, P.O. and Oyama, F.
概要	Acidic mammalian chitinase (AMCase) has been implicated in various pathophysiological conditions including asthma, allergic inflammation and food processing. AMCase is most active at pH 2.0, and its activity gradually decreases to up to pH 8. Here we analyzed chitin degradation by AMCase in weak acidic to neutral conditions by fluorophore-assisted carbohydrate electrophoresis established originally for oligosaccharides analysis. We found that specific fragments with slower-than-expected mobility as defined by chitin oligosaccharide markers were generated at pH 5.0∼8.0 as by-products of the reaction. We established an improved method for chitin oligosaccharides suppressing this side reaction by pre-acidification of the fluorophore-labeling reaction mixture. Our improved method specifically detects chitin oligosaccharides and warrants quantification of up to 50nmol of the material. Using this strategy, we found that AMCase produced dimer of N-acetyl-d-glucosamine (GlcNAc) at strong acidic to neutral condition. Moreover, we found that AMCase generates (GlcNAc)2 as well as (GlcNAc)3 under physiological conditions.
外部リンクURL	https://www.sciencedirect.com/science/article/pii/S0144861717301091?via%3Dihub