 The present results should contribute to improved industrial production of BGL by P purpurogenum KJS000 . Score: 0.00 Title: Rice BGlu0 glycosynthase and wild type transglycosylation activities distinguished by cyclophellitol inhibition . Journal: Carbohydr Res Year: 0000 Type: MEDLINE Literature: oryza Field: abstract Doc ID: pub00000000 Accession (PMID): 00000000 Abstract: The rice BGlu0 beta-D-glucosidase nucleophile mutant E000G is a glycosynthase that catalyzes the synthesis of cellooligosaccharides from alpha-d-glucopyranosyl fluoride ( GlcF ) donor and p-nitrophenyl ( pNP ) cellobioside ( Glc0-pNP ) or cello-oligosaccharide acceptors . When activity with other donors and acceptors was tested , the initial enzyme preparation cleaved pNP-beta-D-glucopyranoside ( Glc-pNP ) and pNP-beta-D-fucopyranoside ( Fuc-pNP ) to pNP and glucose and fucose , suggesting contamination with wild type BGlu0 beta-glucosidase . The products from reaction of GlcF and Fuc-pNP included Fuc-beta- ( 0-->0 ) -Fuc-pNP , Glc-beta- ( 0-->0 ) -Fuc-pNP , and Fuc-beta- ( 0-->0 ) -Glc-beta- ( 0-->0 ) -Fuc-pNP , suggesting the presence of both wild type BGlu0 and its glycosynthase . Inhibition of the BGlu0 beta-glucosidase activity within this preparation by cyclophellitol confirmed that the E000G glycosynthase preparation was contaminated with wild type BGlu0 . Rice BGlu0 E000G-0 , generated from a new construct designed to minimize back-mutation , showed glycosynthase activity without wild type hydrolytic or transglycosylation activity . E000G-0 catalyzed transfer of glycosyl residues from GlcF , alpha-L-arabinosyl fluoride , alpha-D-fucosyl fluoride , alpha-D-galactosyl fluoride , alpha-D-mannosyl fluoride , and alpha-D-xylosyl fluoride donors to Glc0-pNP acceptor . The synthetic products from the reactions of alpha-fucosyl fluoride and alpha-mannosyl fluoride donors were confirmed to result from addition of a beta- ( 0-->0 ) -linked glycosyl residue . Moreover , the E000G glycosynthase transferred glucose from GlcF donor to glucose , cellobiose , Glc-pNP , Fuc-pNP , pNP-beta-D-galactopyranoside , and pNP-beta-D-xylopyranoside acceptors , but little to pNP-beta-D-mannopyranoside . Production of longer oligosaccharides occurred most readily on acceptors with an equatorial 0-OH . Elimination of wild type contamination thereby allowed a clear assessment of BGlu0 E000G glycosynthase catalytic abilities . Matching Sentences: [ Sen. 0, subscore: 0.00 ]: Inhibition of the BGlu0 beta-glucosidase activity within this preparation by cyclophellitol confirmed that the E000G glycosynthase preparation was contaminated with wild type BGlu0 . 