Search tips
Search criteria 


Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. 1995 January; 61(1): 170–174.
PMCID: PMC167272

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


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)

Full Text

The Full Text of this article is available as a PDF (259K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Amato I. The crusade against chlorine. Science. 1993 Jul 9;261(5118):152–154. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Gat O, Lapidot A, Alchanati I, Regueros C, Shoham Y. Cloning and DNA sequence of the gene coding for Bacillus stearothermophilus T-6 xylanase. Appl Environ Microbiol. 1994 Jun;60(6):1889–1896. [PMC free article] [PubMed]
  • Greve LC, Labavitch JM, Hungate RE. alpha-L-arabinofuranosidase from Ruminococcus albus 8: purification and possible role in hydrolysis of alfalfa cell wall. Appl Environ Microbiol. 1984 May;47(5):1135–1140. [PMC free article] [PubMed]
  • Giulian GG, Moss RL, Greaser M. Analytical isoelectric focusing using a high-voltage vertical slab polyacrylamide gel system. Anal Biochem. 1984 Nov 1;142(2):421–436. [PubMed]
  • Hespell RB, O'bryan PJ. Purification and Characterization of an alpha-l-Arabinofuranosidase from Butyrivibrio fibrisolvens GS113. Appl Environ Microbiol. 1992 Apr;58(4):1082–1088. [PMC free article] [PubMed]
  • Khasin A, Alchanati I, Shoham Y. Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6. Appl Environ Microbiol. 1993 Jun;59(6):1725–1730. [PMC free article] [PubMed]
  • Klibanov AM. Stabilization of enzymes against thermal inactivation. Adv Appl Microbiol. 1983;29:1–28. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Manin C, Shareek F, Morosoli R, Kluepfel D. Purification and characterization of an alpha-L-arabinofuranosidase from Streptomyces lividans 66 and DNA sequence of the gene (abfA). Biochem J. 1994 Sep 1;302(Pt 2):443–449. [PubMed]
  • Tajana E, Fiechter A, Zimmermann W. Purification and characterization of two alpha-L-arabinofuranosidases from Streptomyces diastaticus. Appl Environ Microbiol. 1992 May;58(5):1447–1450. [PMC free article] [PubMed]
  • Weinstein L, Albersheim P. Structure of Plant Cell Walls: IX. Purification and Partial Characterization of a Wall-degrading Endo-Arabanase and an Arabinosidase from Bacillus subtilis. Plant Physiol. 1979 Mar;63(3):425–432. [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)