Cellobiose labeled either at the reducing end (4-O-beta-d-glucopyranosyl-d-[U-C]glucopyranose) or at the nonreducing end (4-O-beta-d-[U-C]glucopyranosyl-d -glucopyranose) was synthesized with Clostridium thermocellum cellobiose phosphorylase at greater than 98% purity. The radioactive cellobioses were identical to authentic cellobiose in melting point, optical rotation, isotopic dilution, and c...

Introduction Sugar oxidases and dehydrogenases that catalyze carbohydrate oxidation into the corresponding lactones are of considerable commercial importance. Glucooligosaccharide oxidase (GOOX) from Acremonium strictum was screened with the aim of identifying enzymes with potential applications such as an oligosaccharide acid production and various alternative carbohydrate assays. Screening of...

Characterization in Thermotoga neapolitana of a catabolic gene cluster encoding two glycosyl hydrolases, 1,4-beta-D-glucan glucohydrolase (GghA) and cellobiose phosphorylase (CbpA), and the apparent absence of a cellobiohydrolase (Cbh) suggest a nonconventional pathway for glucan utilization in Thermotogales. GghA purified from T. neapolitana is a 52.5-kDa family 1 glycosyl hydrolase with optim...

The white rot basidiomycete, Phanerochaete chrysosporium, employs an array of extracellular enzymes to completely degrade the major polymers of wood: cellulose, hemicellulose and lignin. Towards the identification of participating enzymes, 268 likely secreted proteins were predicted using SignalP and TargetP algorithms. To assess the reliability of secretome predictions and to evaluate the usef...

The hyperthermophilic archaeon Pyrococcus furiosus can utilize different beta-glucosides, like cellobiose and laminarin. Cellobiose uptake occurs with high affinity (K(m) = 175 nM) and involves an inducible binding protein-dependent transport system. The cellobiose binding protein (CbtA) was purified from P. furiosus membranes to homogeneity as a 70-kDa glycoprotein. CbtA not only binds cellobi...

BACKGROUND The optimization of metabolic pathways is critical for efficient and economical production of biofuels and specialty chemicals. One such significant pathway is the cellobiose utilization pathway, identified as a promising route in biomass utilization. Here we describe the optimization of cellobiose consumption and ethanol productivity by simultaneously engineering both proteins of th...