@article{parker_chhabra_lam_callen_duffaud_snead_short_mathur_kelly_2001, title={Galactomannanases man2 and man5 from Thermotoga species: Growth physiology on galactomannans, gene sequence analysis, and biochemical properties of recombinant enzymes}, volume={75}, ISSN={["0006-3592"]}, DOI={10.1002/bit.10020}, abstractNote={AbstractThe enzymatic hydrolysis of mannan‐based hemicelluloses is technologically important for applications ranging from pulp and paper processing to food processing to gas and oil well stimulation. In many cases, thermostability and activity at elevated temperatures can be advantageous. To this end, the genes encoding β‐mannosidase (man2) and β‐mannanase (man5) from the hyperthermophilic bacteria Thermotoga neapolitana 5068 and Thermotoga maritima were isolated, cloned, and expressed in Escherichia coli. The amino acid sequences for the mannosidases from these organisms were 77% identical and corresponded to proteins with an Mr of approximately 92 kDa. The translated nucleotide sequences for the β‐mannanase genes (man5) encoded polypeptides with an Mr of 76 kDa that exhibited 84% amino acid sequence identity. The recombinant versions of Man2 and Man5 had similar respective biochemical and biophysical properties, which were also comparable to those determined for the native versions of these enzymes in T. neapolitana. The optimal temperature and pH for the recombinant Man2 and Man5 from both organisms were approximately 90°C and 7.0, respectively. The presence of Man2 and Man5 in these two Thermotoga species indicates that galactomannan is a potential growth substrate. This was supported by the fact that β‐mannanase and β‐mannosidase activities were significantly stimulated when T. neapolitana was grown on guar or carob galactomannan. Maximum cell densities increased by at least tenfold when either guar or carob galactomannan was added to the growth medium. For T. neapolitana grown on guar at 83°C, Man5 was secreted into the culture media, whereas Man2 was intracellular. These localizations were consistent with the presence and lack of signal peptides for Man5 and Man2, respectively. The identification of the galactomannan‐degrading enzymes in these Thermotoga species adds to the list of biotechnologically important hemicellulases produced by members of this hyperthermophilic genera. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 322–333, 2001.}, number={3}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Parker, KN and Chhabra, SR and Lam, D and Callen, W and Duffaud, GD and Snead, MA and Short, JM and Mathur, EJ and Kelly, RM}, year={2001}, month={Nov}, pages={322–333} } @article{miller_parker_liebl_lam_callen_snead_mathur_short_kelly_2001, title={alpha-D-galactosidases from Thermotoga species}, volume={330}, journal={Hyperthermophilic enzymes. Part A}, publisher={San Diego, Calif.: Academic Press}, author={Miller, E. S. and Parker, K. N. and Liebl, W. and Lam, D. and Callen, W. and Snead, M. A. and Mathur, E. J. and Short, J. M. and Kelly, R. M.}, year={2001}, pages={246–260} } @article{chang_parker_bauer_kelly_2001, title={alpha-Glucosidase from Pyrococcus furiosus}, volume={330}, journal={Hyperthermophilic enzymes. Part A}, publisher={San Diego, Calif.: Academic Press}, author={Chang, S. T. and Parker, K. N. and Bauer, M. W. and Kelly, R. M.}, year={2001}, pages={260–269} } @article{chhabra_parker_lam_callen_snead_mathur_short_kelly_2001, title={beta-Mannanases from Thermotoga species}, volume={330}, journal={Hyperthermophilic enzymes. Part A}, publisher={San Diego, Calif.: Academic Press}, author={Chhabra, S. and Parker, K. N. and Lam, D. and Callen, W. and Snead, M. A. and Mathur, E. J. and Short, J. M. and Kelly, R. M.}, year={2001}, pages={224–238} } @article{parker_chhabra_lam_snead_mathur_kelly_2001, title={beta-Mannosidase from Thermotoga species}, volume={330}, journal={Hyperthermophilic enzymes. Part A}, publisher={San Diego, Calif.: Academic Press}, author={Parker, K. N. and Chhabra, S. and Lam, D. and Snead, M. A. and Mathur, E. J. and Kelly, R. M.}, year={2001}, pages={238–246} } @article{duffaud_mccutchen_leduc_parker_kelly_1997, title={Purification and characterization of extremely thermostable beta mannanase, beta mannosidase, and alpha galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana}, volume={63}, number={1}, journal={Applied and Environmental Microbiology}, author={Duffaud, G. D. and McCutchen, C. M. and Leduc, P. and Parker, K. N. and Kelly, R. M.}, year={1997}, pages={169–177} }