The functional properties of a xyloglucanase (GH12) of Aspergillus terreus expressed in Aspergillus nidulans may increase performance of biomass degradation
| Autor: | Fernando Segato, André Damasio, Rosymar Coutinho de Lucas, Tony Marcio da Silva, Lucas Ferreira Ribeiro, Marcos Silveira Buckeridge, Cristiane S. Farinas, João Atílio Jorge, Maria de Lourdes Teixeira de Moraes Polizeli, Aline Zorzetto Lopes Gonçalves, Gabriela Leal Vitcosque, Liliane Fraga Costa Ribeiro |
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| Rok vydání: | 2015 |
| Předmět: |
0106 biological sciences
0301 basic medicine Glycoside Hydrolases Lignocellulosic biomass Enzyme Activators Gene Expression Context (language use) 01 natural sciences Applied Microbiology and Biotechnology Lignin Microbiology Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Bioreactors Aspergillus nidulans 010608 biotechnology Enzyme Stability Tamarindus Aspergillus terreus Cloning Molecular Enzyme Inhibitors biology Temperature Substrate (chemistry) General Medicine POLISSACARÍDEOS Hydrogen-Ion Concentration biology.organism_classification Recombinant Proteins Xyloglucan 030104 developmental biology Aspergillus chemistry Biochemistry Fermentation Xylanase Biotechnology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 1432-0614 |
| Popis: | Filamentous fungi are attractive hosts for heterologous protein expression due to their capacity to secrete large amounts of enzymes into the extracellular medium. Xyloglucanases, which specifically hydrolyze xyloglucan, have been recently applied in lignocellulosic biomass degradation and conversion in many other industrial processes. In this context, this work aimed to clone, express, and determine the functional properties of a recombinant xyloglucanase (AtXEG12) from Aspergillus terreus, and also its solid-state (SSF) and submerged (SmF) fermentation in bioreactors. The purified AtXEG12 showed optimum pH and temperature of 5.5 and 65 °C, respectively, demonstrating to be 90 % stable after 24 h of incubation at 50 °C. AtXEG12 activity increased in the presence of 2-mercaptoethanol (65 %) and Zn+2 (45 %), while Cu+2 and Ag+ ions drastically decreased its activity. A substrate assay showed, for the first time for this enzyme’s family, xylanase activity. The enzyme exhibited high specificity for tamarind xyloglucan (K M 1.2 mg mL−1) and V max of 17.4 μmol min−1 mg−1 of protein. The capillary zone electrophoresis analysis revealed that AtXEG12 is an endo-xyloglucanase. The heterologous xyloglucanase secretion was greater than the production by wild-type A. terreus cultivated in SmF. On the other hand, AtXEG12 activity reached by SSF was sevenfold higher than values achieved by SmF, showing that the expression of recombinant enzymes can be significantly improved by cultivation under SSF. |
| Databáze: | OpenAIRE |
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