EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Purification and characterization of a thermostable beta xylosidase from thermomonospora fusca



Purification and characterization of a thermostable beta xylosidase from thermomonospora fusca



Journal of General Microbiology 135(2): 293-300



Xylan-degrading enzymes, including .beta.-xylosidase (EC 3.2.1.37), were induced when Thermomonospora fusca was grown at 50.degree. C in liquid medium containing 0.2% xylan. The intracellular .beta.-xylosidase activity was concentrated and characterized by fast protein liquid chromatography and gel electrophoresis. A zymogram technique was developed to identify .beta.-xylosidase directly on polyacrylamide gels. A single enzyme (168 kDa; pI 4.37) was identified and purified to homogeneity. The consistent detection of a single band on denaturing SDS gels suggested that the enzyme was composed of identical subunits; since the subunit molecular mass was 56 kDa, a trimeric is suggested. High activity against p-nitrophenyl .beta.-D-xylopyranoside (pNPX) occurred in the pH range 5.0-9.0 and temperature range 40-60.degree. C. The enzyme was stable at room temperature at pH 6.0-8.0; it had a half-life of 8 h at 65.degree. C, and of 1.5 h at 70.degree. C. The purified enzyme did not exhibit any detectable activity against arabinoxylan, carboxymethylcellulose or p-nitrophenyl .beta.-D-glucopyranoside. The enzyme had a Km of 0.89 mM (pNPX) and was inhibited by D-xylose (Ki 19 mM) but not D-glucose. The size of the T. fusca enzyme is in the range reported for the few other bacterial .beta.-xylosidases described, but the acidic nature of the protein and its affinity for the substrate have more in common with some of the monomeric .beta.-xylosidases described in fungi.

(PDF emailed within 1 workday: $29.90)

Accession: 007707185

Download citation: RISBibTeXText



Related references

Purification and characterization of a thermostable b-xylosidase from Thermomonospora fusca. The Journal of General Microbiology 135: 3-9, 1989

Thermostable beta-glucosidase from Thermomonospora fusca: Purification and biochemical characterization. Agricultural Chemistry & Biotechnology 46(1): 6-11, March, 2003

Characterization of beta-xylosidase and alpha-L-arabinofuranosidase activities from Thermomonospora fusca BD25. Turkish Journal of Biology 24(4): 753-767, 2000

Thermostable beta-xylosidase from Thermomonospora curvata. Journal of Industrial Microbiology & Biotechnology 20(1): 55-60, 1998

Purification and characterization of two beta 1 4 endoglucanases from thermomonospora fusca. Biochemistry 24(26): 7797-7804, 1985

Purification and characterization of a thermostable beta-xylosidase from Thermoanaerobacter ethanolicus. Journal of Bacteriology 174(18): 5848-5853, 1992

Purification and characterization of thermostable xylanase and beta-xylosidase by the thermophilic bacterium Bacillus thermantarcticus. Research in Microbiology 155(4): 283-289, 2004

Purification and characterization of a thermostable intracellular beta-xylosidase from the thermophilic fungus Sporotrichum thermophile. Process Biochemistry 41(12): 2402-2409, 2006

Expression and characterization of a thermostable serine protease (TfpA) from Thermomonospora fusca YX in Pichia pastoris. Applied Microbiology and Biotechnology 68(3): 355-359, 2005

Purification and characterization of two b-1,4-endoglucanases from Thermomonospora fusca. Biochemistry (American Chemical Society) 24: 97-804, 1985

High-resolution native and complex structures of thermostable beta-mannanase from Thermomonospora fusca - substrate specificity in glycosyl hydrolase family 5. Structure 6(11): 1433-1444, 1998

Purification and characterization of the heat-stable serine proteinase from Thermomonospora fusca YX. Biochemical Journal 246(2): 511-517, 1987

Purification and partial characterization of alpha-L-arabinofuranosidase produced by Thermomonospora fusca. Folia Microbiologica 48(2): 168-172, 2003

Purification and characterization of a recombinant β-D-xylosidase from Thermobifida fusca TM51. Protein Journal 31(8): 641-650, 2013

Purification and characterization of a thermostable b-xylosidase from Thermoanaerobacter ethanolicus. Journal of Bacteriology 174(18): 48-53, 1992