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Appl Microbiol Biotechnol. 2016 Nov;100(21):9133-9144. Epub 2016 Jun 1.

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

Author information

1
Departamento de Bioquímica e Imunologia, FMRP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
2
Chemical Biochemical Environmental Engineering Department, University of Maryland, Baltimore County, MD, USA.
3
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes, 3900, Ribeirão Preto, SP, 14040-901, Brazil.
4
Instituto de Ciências da Saúde, Agrárias e Humanas do Centro Universitário do Planalto de Araxà (UNIARAXÀ), Araxà, MG, Brazil.
5
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA.
6
Departamento de Bioquímica e Biologia Tecidual, Instituto de Biologia, Universidade de Campinas, Campinas, SP, Brazil.
7
Empresa Brasileira de Pesquisa Agropecuária, EMBRAPA Instrumentação, São Carlos, SP, Brazil.
8
Departamento de Biotecnologia, Escola de Engenharia de Lorena, Universidade de São Paulo, São Paulo, SP, Brazil.
9
Departamento de Botanica, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil.
10
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes, 3900, Ribeirão Preto, SP, 14040-901, Brazil. polizeli@ffclrp.usp.br.

Abstract

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.

KEYWORDS:

Aspergillus terreus; Recombinant enzymes; SSF; SmF; Xyloglucanases

PMID:
27245677
DOI:
10.1007/s00253-016-7589-2
[Indexed for MEDLINE]

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