RRC ID 50858
Author Taguchi Y, Saburi W, Imai R, Mori H.
Title Evaluation of acceptor selectivity of Lactococcus lactis ssp. lactis trehalose 6-phosphate phosphorylase in the reverse phosphorolysis and synthesis of a new sugar phosphate.
Journal Biosci Biotechnol Biochem
Abstract Trehalose 6-phosphate phosphorylase (TrePP), a member of glycoside hydrolase family 65, catalyzes the reversible phosphorolysis of trehalose 6-phosphate (Tre6P) with inversion of the anomeric configuration to produce β-d-glucose 1-phosphate (β-Glc1P) and d-glucose 6-phosphate (Glc6P). TrePP in Lactococcus lactis ssp. lactis (LlTrePP) is, alongside the phosphotransferase system, involved in the metabolism of trehalose. In this study, recombinant LlTrePP was produced and characterized. It showed its highest reverse phosphorolytic activity at pH 4.8 and 40°C, and was stable in the pH range 5.0-8.0 and at up to 30°C. Kinetic analyses indicated that reverse phosphorolysis of Tre6P proceeded through a sequential bi bi mechanism involving the formation of a ternary complex of the enzyme, β-Glc1P, and Glc6P. Suitable acceptor substrates were Glc6P, and, at a low level, d-mannose 6-phosphate (Man6P). From β-Glc1P and Man6P, a novel sugar phosphate, α-d-Glcp-(1↔1)-α-d-Manp6P, was synthesized with 51% yield.
Volume 81(8)
Pages 1512-1519
Published 2017-8-1
DOI 10.1080/09168451.2017.1329620
PMID 28537141
MeSH Bacterial Proteins / genetics Bacterial Proteins / metabolism* Cloning, Molecular Escherichia coli / genetics Escherichia coli / metabolism Gene Expression Glucosephosphates / metabolism Glucosyltransferases / genetics Glucosyltransferases / metabolism* Hydrogen-Ion Concentration Hydrolysis Kinetics Lactococcus lactis / chemistry Lactococcus lactis / enzymology* Mannosephosphates / metabolism Recombinant Proteins / genetics Recombinant Proteins / metabolism Substrate Specificity Sugar Phosphates / biosynthesis* Sugar Phosphates / metabolism Temperature Trehalose / analogs & derivatives* Trehalose / metabolism*
IF 1.516
Times Cited 3
General Microbes JCM 5805