Mass spectrometric analysis using ruthenium (II)-labeling for identification of glycosyl hydrolase product.
Analysis of products digested by glycosyl hydrolases helps understanding of the hydrolysis mechanism and the substrate recognition in the enzymes. We developed a new universal technique, which consists of ruthenium (II) complex labeling and mass spectrometry analysis, to identify the reducing sugars released from oligosaccharides by enzymatic digestion. This method was applied to enzymatic digestion by chitinase and cellulase of the hyperthermophilic archaea Pyrococcus fusiosus and Pyrococcus horikoshii respectively.
Ito
Akihiro
A
National Institute of Advanced Industrial Science and Technology, Ikeda, Osaka, Japan.
Okamura
Taka-aki
TA
Uegaki
Koichi
K
Kim
Han-Woo
HW
Ishikawa
Kazuhiko
K
Nakamura
Tsutomu
T
Yamamoto
Hitoshi
H
Ueyama
Norikazu
N
D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry; D002480:Cellulase / Q000378:metabolism; D002688:Chitinases / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D009942:Organometallic Compounds / Q000737:chemistry; D020027:Pyrococcus furiosus / Q000201:enzymology; D043902:Pyrococcus horikoshii / Q000201:enzymology; D012428:Ruthenium / Q000032:analysis* / Q000737:chemistry; D013194:Staining and Labeling
D002241:Carbohydrates; D009942:Organometallic Compounds; D012428:Ruthenium; D002688:Chitinases; D002480:Cellulase; D009699:N-Glycosyl Hydrolases