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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.)

Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation

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Author(s):
Trindade, Lucas Vinicius ; Desagiacomo, Carla ; Teixeira de Moraes Polizeli, Maria de Lourdes ; de Lima Damasio, Andre Ricardo ; Furuyama Lima, Aline Margarete ; Gomes, Eleni ; Bonilla-Rodriguez, Gustavo Orlando
Total Authors: 7
Document type: Journal article
Source: BIOMED RESEARCH INTERNATIONAL; 2016.
Web of Science Citations: 2
Abstract

This work reports the production of an exo-polygalacturonase (exo-PG) by Rhizomucor pusillus A13.36 in submerged cultivation (SmC) in a shaker at 45 degrees C for 96 h. A single pectinase was found and purified in order to analyze its thermal stability, by salt precipitation and hydrophobic interaction chromatography. The pectinase has an estimated Mw of approximately 43.5-47 kDa and optimum pH of 4.0 but is stable in pH ranging from 3.5 to 9.5 and has an optimum temperature of 61 degrees C. It presents thermal stability between 30 and 60 degrees C, has 70% activation in the presence of Ca2+, and was tested using citrus pectin with a degree of methyl esterification (DE) of 26% E-a(d) for irreversible denaturation was 125.5 kJ/mol with positive variations of entropy and enthalpy for that and Delta G((d)) values were around 50 kJ/mol. The hydrolysis of polygalacturonate was analyzed by capillary electrophoresis which displayed a pattern of sequential hydrolysis (exo). The partial identification of the primary sequence was done by MS MALDI-TOF and a comparison with data banks showed the highest identity of the sequenced fragments of exo-PG from R. pusillus with an exo-pectinase from Aspergillus fumigatus. Pectin hydrolysis showed a sigmoidal curve for the Michaelis-Menten plot. (AU)

FAPESP's process: 10/12624-0 - Application of physical-chemical and enzymatic methods in the saccharification of sugar cane bagasse: studies on microorganisms, fermentative processes and hydrolyses methods
Grantee:Eleni Gomes
Support type: Research Projects - Thematic Grants
FAPESP's process: 11/23991-7 - Functional and structural analysis of microbial and plant hydrolases with potential use in biotechnological or industrial processes
Grantee:Gustavo Orlando Bonilla Rodriguez
Support type: Regular Research Grants