This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Duffaud, G. D. Articles by Kelly, R. M. Search for Related Content PubMed PubMed Citation Articles by Duffaud, G. D. Articles by Kelly, R. M. Agricola Articles by Duffaud, G. D. Articles by Kelly, R. M.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 01 1997, 169-177, Vol 63, No. 1
Copyright © 1997, American Society for Microbiology

Purification and characterization of extremely thermostable beta- mannanase, beta-mannosidase, and alpha-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana 5068

GD Duffaud, CM McCutchen, P Leduc, KN Parker and RM Kelly
Department of Chemical Engineering, North Carolina State University, Raleigh 27695-7905, USA.

Thermostable and thermoactive beta-mannanase (1,4-beta-D-mannan mannanohydrolase [EC 3.2.1.78]), beta-mannosidase (beta-D- mannopyranoside hydrolase [EC 3.2.1.25]) and alpha-galactosidase (alpha- D-galactoside galactohydrolase [EC 3.2.1.22]) were purified to homogeneity from cell extracts and extracellular culture supernatants of the hyperthermophilic eubacterium Thermotoga neapolitana 5068 grown on guar gum-based media. The beta-mannanase was an extracellular monomeric enzyme with a molecular mass of 65 kDa. The optimal temperature for activity was 90 to 92 degrees C, with half-lives (t1/2) of 34 h at 85 degrees C, 13 h at 90 degrees C, and 35 min at 100 degrees C. The beta-mannosidase and alpha-galactosidase were found primarily in cell extracts. The beta-mannosidase was a homodimer consisting of approximately 100-kDa molecular mass subunits. The optimal temperature for activity was 87 degrees C, with t1/2 of 18 h at 85 degrees C, 42 min at 90 degrees C, and 2 min at 98 degrees C. The alpha-galactosidase was a 61-kDa monomeric enzyme with a temperature optimum of 100 to 103 degrees C and t1/2 of 9 h at 85 degrees C, 2 h at 90 degrees C, and 3 min at 100 degrees C. These enzymes represent the most thermostable and thermoactive versions of these types yet reported and probably act synergistically to hydrolyze extracellular galactomannans to monosaccharides by T. neapolitana for nutritional purposes. The significance of such substrates in geothermal environments remains to be seen.

This article has been cited by other articles:

• Beki, E., Nagy, I., Vanderleyden, J., Jager, S., Kiss, L., Fulop, L., Hornok, L., Kukolya, J. (2003). Cloning and Heterologous Expression of a {beta}-D-Mannosidase (EC 3.2.1.25)-Encoding Gene from Thermobifida fusca TM51. Appl. Environ. Microbiol. 69: 1944-1952 [Abstract] [Full Text]  
• Chhabra, S. R., Shockley, K. R., Ward, D. E., Kelly, R. M. (2002). Regulation of Endo-Acting Glycosyl Hydrolases in the Hyperthermophilic Bacterium Thermotoga maritima Grown on Glucan- and Mannan-Based Polysaccharides. Appl. Environ. Microbiol. 68: 545-554 [Abstract] [Full Text]  
• Ishiguro, M., Kaneko, S., Kuno, A., Koyama, Y., Yoshida, S., Park, G.-G., Sakakibara, Y., Kusakabe, I., Kobayashi, H. (2001). Purification and Characterization of the Recombinant Thermus sp. Strain T2 {alpha}-Galactosidase Expressed in Escherichia coli. Appl. Environ. Microbiol. 67: 1601-1606 [Abstract] [Full Text]  
• Yernool, D. A., McCarthy, J. K., Eveleigh, D. E., Bok, J.-D. (2000). Cloning and Characterization of the Glucooligosaccharide Catabolic Pathway beta -Glucan Glucohydrolase and Cellobiose Phosphorylase in the Marine Hyperthermophile Thermotoga neapolitana. J. Bacteriol. 182: 5172-5179 [Abstract] [Full Text]  
• Sunna, A., Gibbs, M. D., Chin, C. W. J., Nelson, P. J., Bergquist, P. L. (2000). A Gene Encoding a Novel Multidomain beta -1,4-Mannanase from Caldibacillus cellulovorans and Action of the Recombinant Enzyme on Kraft Pulp. Appl. Environ. Microbiol. 66: 664-670 [Abstract] [Full Text]  
• Stoll, D., Stålbrand, H., Warren, R. A. J. (1999). Mannan-Degrading Enzymes from Cellulomonas fimi. Appl. Environ. Microbiol. 65: 2598-2605 [Abstract] [Full Text]  
• Bok, J.-D., Yernool, D. A., Eveleigh, D. E. (1998). Purification, Characterization, and Molecular Analysis of Thermostable Cellulases CelA and CelB from Thermotoga neapolitana. Appl. Environ. Microbiol. 64: 4774-4781 [Abstract] [Full Text]