The functional properties of a xyloglucanase (GH12) of Aspergillus terreus expressed in Aspergillus nidulans may increase performance of biomass degradation

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Vitcosque, Gabriela Leal ; Ribeiro, Liliane Fraga Costa ; de Lucas, Rosymar Coutinho ; da Silva, Tony Marcio ; Ribeiro, Lucas Ferreira ; de Lima Damasio, Andre Ricardo ; Farinas, Cristiane Sanchez ; Lopes Goncalves, Aline Zorzetto ; Segato, Fernando ; Buckeridge, Marcos Silveira ; Jorge, Joao Atilio ; Polizeli, Maria de Lourdes T. M.

• Tipo de documento: Artigo Científico
• Fonte: Applied Microbiology and Biotechnology; v. 100, n. 21, p. 9133-9144, NOV 2016.
• Citações Web of Science: 3

Resumo

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 A degrees C, respectively, demonstrating to be 90 % stable after 24 h of incubation at 50 A degrees 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 mu mol 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. (AU)

• Processo FAPESP: 10/10184-3 - Desenvolvimento de um coquetel enzimático para aplicação em biorefinaria
• Beneficiário: Liliane Fraga Costa Ribeiro
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/18714-2 - Oxidação enzimática do bagaço de cana-de-açúcar: descoberta, caracterização e aplicação de novas enzimas oxidativas de carboidratos, voltadas para o desenvolvimento de um cell factory mais eficiente
• Beneficiário: Fernando Segato
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Jovens Pesquisadores
• Processo FAPESP: 12/20549-4 - Secreção de glicoproteínas heterólogas em Aspergillus: efeito do padrão de glicosilação em parâmetros funcionais de glicosil hidrolases
• Beneficiário: André Ricardo de Lima Damasio
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Jovens Pesquisadores
• Processo FAPESP: 10/52322-3 - Bioprospecção de fungos filamentosos produtores de holoenzimas com aplicação em biorefinaria
• Beneficiário: Maria de Lourdes Teixeira de Moraes Polizeli
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOTA - Regular