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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.)

Biophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.

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Rosseto, Flavio Rodolfo ; Manzine, Livia Regina ; Neto, Mario de Oliveira ; Polikarpov, Igor
Total Authors: 4
Document type: Journal article
Source: Enzyme and Microbial Technology; v. 91, p. 1-7, SEP 2016.
Web of Science Citations: 2

Endoglucanases are the main cellulolytic enzymes secreted by the bacterium Xanthomonas campestris pv. campestris (Xcc). The major endoglucanase exported by this bacterium into an external milieu is an enzyme XccCel5A, which belongs to GH5 family subfamily 1 and is encoded by the gene engXCA. We purified XccCel5A using ammonium sulfate precipitation followed by size exclusion chromatography and identified it by zymogram analysis. Circular dichroism and fluorescence spectroscopy studies showed that XccCel5A is stable in a wide pH range and up to about 55 degrees C and denatures at the higher temperatures. The optimal conditions for enzyme activity were identified as T = 45 degrees C and pH = 7.0. Under the optimum conditions the catalytic efficiency (k(cat)/K-M) of the enzyme was determined as 5.16 x 10(4)s(-1) M-1 using carboxymethylcellulose (CMC) as a substrate. Our SAXS studies revealed extended tadpole-shape molecular assembly, typical for cellulases, and allowed to determine an overall shape of the enzyme and a relative position of the catalytic and cellulose binding domains. (C) 2016 Elsevier Inc. All rights reserved. (AU)

FAPESP's process: 09/54035-4 - Facility for advanced studies of biosystems and nanostructured materials
Grantee:Igor Polikarpov
Support type: Multi-user Equipment Program
FAPESP's process: 10/16542-9 - Studies of the cellulolytic enzymes with potential in the enzymatic transformation of the sugarcane biomass using Small Angle X-ray Scattering
Grantee:Mario de Oliveira Neto
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 10/08370-3 - Biophysical and biochemical studies of exoglucanases from Trichoderma harzianum involved in the biodegradation of cellulose
Grantee:Wanius José Garcia da Silva
Support type: Regular Research Grants
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 type: Program for Research on Bioenergy (BIOEN) - Thematic Grants