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

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Jan 1995, 170-174, Vol 61, No. 1
Copyright © 1995, American Society for Microbiology

Purification and characterization of alpha-L-arabinofuranosidase from Bacillus stearothermophilus T-6

S Gilead and Y Shoham
Department of Food Engineering and Biotechnology, Technion-Israel Institute of Technology, Haifa.

Bacillus stearothermophilus T-6 produced an alpha-L-arabinofuranosidase when grown in the presence of L-arabinose, sugar beet arabinan, or oat spelt xylan. At the end of a fermentation, about 40% of the activity was extracellular, and enzyme activity in the cell-free supernatant could reach 25 U/ml. The enzymatic activity in the supernatant was concentrated against polyethylene glycol 20000, and the enzyme was purified eightfold by anion-exchange and hydrophobic interaction chromatographies. The molecular weight of T-6 alpha-L- arabinofuranosidase was 256,000, and it consisted of four identical subunits as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration. The native enzyme had a pI of 6.5 and was most active at 70 degrees C and at pH 5.5 to 6.0. Its thermostability at pH 7.0 was characterized by half-lives of 53, 15, and 1 h at 60, 65, and 70 degrees C, respectively. Kinetic experiments at 60 degrees C with p-nitrophenyl alpha-L-arabinofuranoside as a substrate gave a Vmax, a Km, and an activation energy of 749 U/mg, 0.42 mM, and 16.6 kcal/mol, (ca. 69.5 kJ/mol), respectively. The enzyme had no apparent requirement for cofactors, and its activity was strongly inhibited by 1 mM Hg2+. T-6 alpha-L-arabinofuranosidase released L- arabinose from arabinan and had low activity on oat spelt xylan. The enzyme acted cooperatively with T-6 xylanase in hydrolyzing oat spelt xylan, and L-arabinose, xylose, and xylobiose were detected as the end reaction products.(ABSTRACT TRUNCATED AT 250 WORDS)


This article has been cited by other articles:

  • Inacio, J. M., Correia, I. L., de Sa-Nogueira, I. (2008). Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis. Microbiology 154: 2719-2729 [Abstract] [Full Text]  
  • Bauer, S., Vasu, P., Persson, S., Mort, A. J., Somerville, C. R. (2006). From the Cover: Development and application of a suite of polysaccharide-degrading enzymes for analyzing plant cell walls. Proc. Natl. Acad. Sci. USA 103: 11417-11422 [Abstract] [Full Text]  
  • Raposo, M. P., Inacio, J. M., Mota, L. J., de Sa-Nogueira, I. (2004). Transcriptional Regulation of Genes Encoding Arabinan-Degrading Enzymes in Bacillus subtilis. J. Bacteriol. 186: 1287-1296 [Abstract] [Full Text]  
  • Golan, G., Shallom, D., Teplitsky, A., Zaide, G., Shulami, S., Baasov, T., Stojanoff, V., Thompson, A., Shoham, Y., Shoham, G. (2004). Crystal Structures of Geobacillus stearothermophilus {alpha}-Glucuronidase Complexed with Its Substrate and Products: MECHANISTIC IMPLICATIONS. J. Biol. Chem. 279: 3014-3024 [Abstract] [Full Text]  
  • Margolles, A., de los Reyes-Gavilan, C. G. (2003). Purification and Functional Characterization of a Novel {alpha}-L-Arabinofuranosidase from Bifidobacterium longum B667. Appl. Environ. Microbiol. 69: 5096-5103 [Abstract] [Full Text]  
  • Kosugi, A., Murashima, K., Doi, R. H. (2002). Characterization of Two Noncellulosomal Subunits, ArfA and BgaA, from Clostridium cellulovorans That Cooperate with the Cellulosome in Plant Cell Wall Degradation. J. Bacteriol. 184: 6859-6865 [Abstract] [Full Text]  
  • Shallom, D., Belakhov, V., Solomon, D., Shoham, G., Baasov, T., Shoham, Y. (2002). Detailed Kinetic Analysis and Identification of the Nucleophile in alpha -L-Arabinofuranosidase from Geobacillus stearothermophilus T-6, a Family 51 Glycoside Hydrolase. J. Biol. Chem. 277: 43667-43673 [Abstract] [Full Text]  
  • Debeche, T., Cummings, N., Connerton, I., Debeire, P., O'Donohue, M. J. (2000). Genetic and Biochemical Characterization of a Highly Thermostable alpha -L-Arabinofuranosidase from Thermobacillus xylanilyticus. Appl. Environ. Microbiol. 66: 1734-1736 [Abstract] [Full Text]  
  • Lyon, P.-F., Beffa, T., Fischer, J. L., Aragno, M. (2000). Xylanase activity and thermostratification during the thermogenic phase of industrial composting in aerated trenches. Waste Manag Res 18: 174-183 [Abstract]  
  • Shulami, S., Gat, O., Sonenshein, A. L., Shoham, Y. (1999). The Glucuronic Acid Utilization Gene Cluster from Bacillus stearothermophilus T-6. J. Bacteriol. 181: 3695-3704 [Abstract] [Full Text]  
  • Ethier, N., Talbot, G., Sygusch, J. (1998). Gene Cloning, DNA Sequencing, and Expression of Thermostable beta -Mannanase from Bacillus stearothermophilus. Appl. Environ. Microbiol. 64: 4428-4432 [Abstract] [Full Text]  
  • Saha, B. C., Bothast, R. J. (1998). Purification and Characterization of a Novel Thermostable alpha -L-Arabinofuranosidase from a Color-Variant Strain of Aureobasidium pullulans. Appl. Environ. Microbiol. 64: 216-220 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»