Crystallization and preliminary diffraction analysis of the catalytic domain of major extracellular endoglucanase from Xanthomonas campestris pv. campestris

Author(s)

Rosseto, Flavio R. ; Puhl, Ana C. ; Andrade, Maxuel O. ; Polikarpov, Igor

• Document type: Journal article
• Source: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS; v. 69, p. 4-pg., 2013-02-01.

Abstract

Cellulases, such as endoglucanases, exoglucanases and beta-glucosidases, are important enzymes used in the process of enzymatic hydrolysis of plant biomass. The bacteria Xanthomonas campestris pv. campestris expresses a large number of hydrolases and the major endoglucanase (XccEG), a member of glycoside hydrolase family 5 (GH5), is the most strongly secreted extracellularly. In this work, the native XccEG was purified from the extracellular extract and crystallization assays were performed on its catalytic domain. A complete data set was collected on an in-house X-ray source. The crystal diffracted to 2.7 angstrom resolution and belonged to space group C2, with unit-cell parameters a = 174.66, b = 141.53, c = 108.00 angstrom, beta = 110.49 degrees. The Matthews coefficient suggests a solvent content of 70.1% and the presence of four protein subunits in the asymmetric unit. (AU)

• FAPESP's process: 10/52362-5 - Targeted analysis of microbial lignocellulolytic secretomes: a new approach to enzyme discovery
• Grantee: Igor Polikarpov
• Support Opportunities: Regular Research Grants
• FAPESP's process: 09/05349-6 - Expression and purification of cellobiohydrolase I from the filamentous fungus Trichoderma harzianum
• Grantee: Bruno Luan Soares Paula de Mello
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 08/56255-9 - Structure and function of enzymes and auxiliary proteins from Trichoderma, active in cell-wall hydrolysis
• Grantee: Igor Polikarpov
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Thematic Grants
• FAPESP's process: 07/08706-9 - Molecular and Structural Studies of Cellulitic Complex Enzymes of Fungi Trichoderma harzianum with Potencial of Enzymatic Transformation of Biomass
• Grantee: Viviane Isabel Serpa
• Support Opportunities: Scholarships in Brazil - Doctorate