Acceptor specificity of trehalose phosphorylase from Thermoanaerobacter brockii: production of novel nonreducing trisaccharide, 6-O-alpha-D-galactopyranosyl trehalose.
We investigated the acceptor specificity of a thermostable trehalose phosphorylase from Thermoanaerobacter brockii ATCC 35047 (TbTP) was examined using beta-D-glucose-1-phosphate (beta-G1P) as a glucosyl donor and oligosaccharides as the acceptor. Oligosaccharides with a reducing-end glucose residue as the C-6 substituent (e.g., isomaltose, gentiobiose, melibiose, isomaltotriose, and isopanose) were found to be successful acceptors. The transfer products of isomaltose, gentiobiose, and melibiose were isolated and characterized as 6-O-alpha-D-glucopyranosyl trehalose (alpha-GlcTre), 6-O-beta-D-glucopyranosyl trehalose (beta-GlcTre), and 6-O-alpha-D-galactopyranosyl trehalose (alpha-GalTre), respectively. To produce alpha-GalTre, a novel nonreducing trisaccharide, the reaction conditions of alpha-GalTre were examined using trehalose as a glucosyl donor. As a result, the yield of alpha-GalTre reached 40.5%.
Maruta
Kazuhiko
K
Amase Institute, Hayashibara Biochemical Laboratories, Inc., Okayama, Japan. kmaruta@hayashibara.co.jp
Watanabe
Hikaru
H
Nishimoto
Tomoyuki
T
Kubota
Michio
M
Chaen
Hiroto
H
Fukuda
Shigeharu
S
Kurimoto
Masashi
M
Tsujisaka
Yoshio
Y
D004789:Enzyme Activation; D004795:Enzyme Stability; D005947:Glucose / Q000737:chemistry*; D005958:Glucosephosphates / Q000737:chemistry*; D005964:Glucosyltransferases / Q000737:chemistry*; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D013696:Temperature; D046968:Thermoanaerobacter / Q000201:enzymology*; D014312:Trisaccharides / Q000737:chemistry*
D005958:Glucosephosphates; D014312:Trisaccharides; C031590:glucose-1-phosphate; D005964:Glucosyltransferases; C023181:alpha,alpha-trehalose phosphorylase; D005947:Glucose