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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Molecular cloning, overexpression, purification and crystallographic analysis of a GH43-xylosidase from Bacillus licheniformis

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Author(s):
Diogo, Jose Alberto [1] ; Zanphorlin, Leticia Maria [1] ; Sato, Helia Harumi [2] ; Murakami, Mario Tyago [3] ; Ruller, Roberto [1]
Total Authors: 5
Affiliation:
[1] Natl Ctr Res Energy & Mat CNPEM, Brazilian Bioethanol Sci & Technol Lab CTBE, BR-13083100 Campinas, SP - Brazil
[2] Univ Estadual Campinas, Dept Food Sci, BR-13081970 Campinas, SP - Brazil
[3] Natl Ctr Res Energy & Mat CNPEM, Brazilian Biosci Natl Lab LNBio, BR-13083100 Campinas, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 71, n. 8, p. 962-965, AUG 2015.
Web of Science Citations: 1
Abstract

-Xylosidases (EC 3.2.1.37) catalyze the hydrolysis of short xylooligosaccharides into xylose, which is an essential step in the complete depolymerization of xylan, the major hemicellulosic polysaccharide of plant cell walls, and has great biotechnological relevance for the production of lignocellulose-based biofuels and the paper industry. In this study, a GH43 -xylosidase identified from the bacterium Bacillus licheniformis (BlXylA) was cloned into the the pET-28a bacterial expression vector, recombinantly overexpressed in Escherichia coli BL21(DE3) cells and purified to homogeneity by metal-affinity and size-exclusion chromatography. The protein was crystallized in the presence of the organic solvent 2-methyl-2,4-pentanediol and a single crystal diffracted to 2.49 angstrom resolution. The X-ray diffraction data were indexed in the monoclinic space group C2, with unit-cell parameters a = 152.82, b = 41.9, c = 71.79 angstrom, = 91.7 degrees. Structural characterization of this enzyme will contribute to a better understanding of the structural requirements for xylooligosaccharide specificity within the GH43 family. (AU)

FAPESP's process: 13/13309-0 - Studies of the structural and functional behavior of enzymes evolutionarily specialized in the degradation of plant biomass with potential biotechnological applications
Grantee:Mário Tyago Murakami
Support type: Regular Research Grants
FAPESP's process: 13/10443-7 - Bioprospecting and construction of hemicellulases for the production of xylose from hydrolyzate licor pentoses and their use in fermentative xylitol production
Grantee:José Alberto Diogo
Support type: Scholarships in Brazil - Master
FAPESP's process: 14/07135-1 - Multi-user equipament approved in grant 2013/13309-0: Beckman Coulter capillary electrophoresis systems with LIF detection system
Grantee:Mário Tyago Murakami
Support type: Multi-user Equipment Program