title: Cloning, heterologous expression and biochemical characterization of a non-specific endoglucanase family 12 from Aspergillus terreus NIH2624.
abstract: The cellulases from Glycoside Hydrolyses family 12 (GH12) play an important role in cellulose degradation and plant cell wall deconstruction being widely used in a number of bioindustrial processes. Aiming to contribute toward better comprehension of these class of the enzymes, here we describe a high-yield secretion of a endoglucanase GH12 from Aspegillus terreus (AtGH12), which was cloned and expressed in Aspergillus nidulans strain A773. The purified protein was used for complete biochemical and functional characterization. The optimal temperature and pH of the enzyme were 55°C and 5.0 respectively, which has high activity against β-glucan and xyloglucan and also is active toward glucomannan and CMC. The enzyme retained activity up to 60°C. AtGH12 is strongly inhibited by Cu2+, Fe2+, Cd2+, Mn2+, Ca2+, Zn2+and EDTA, whereas K+, Tween, Cs+, DMSO, Triton X-100 and Mg2+enhanced the enzyme activity. Furthermore, SAXS data reveal that the enzyme has a globular shape and CD analysis demonstrated a prevalence of a β-strand structure corroborating with typical β-sheets fold commonly found for other endoglucanases from GH12 family.
authors.affiliation: Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil; Departamento de Biotecnologia, Escola de Engenharia de Lorena - Universidade de São Paulo, Lorena, SP, Brazil.
authors.forename: Fernando
authors.initials: F
authors.lastname: Segato
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Bruno
authors.initials: B
authors.lastname: Dias
authors.affiliation: Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil; Departamento de Biotecnologia, Escola de Engenharia de Lorena - Universidade de São Paulo, Lorena, SP, Brazil.
authors.forename: Gabriela L
authors.initials: GL
authors.lastname: Berto
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Dyoni M
authors.initials: DM
authors.lastname: de Oliveira
authors.affiliation: Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Flávio H M
authors.initials: FHM
authors.lastname: De Souza
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Citadini
authors.affiliation: Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Mario T
authors.initials: MT
authors.lastname: Murakami
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: André R L
authors.initials: ARL
authors.lastname: Damásio
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP, Brazil.
authors.forename: Fábio Márcio
authors.initials: FM
authors.lastname: Squina
authors.affiliation: Departamento de Física e Informática, Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil. Electronic address: ipolikarpov@ifsc.usp.br.
authors.forename: Igor
authors.initials: I
authors.lastname: Polikarpov
mesh_terms: D001230:Aspergillus / Q000201:enzymology / Q000235:genetics; D002480:Cellulase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D011994:Recombinant Proteins
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D002480:Cellulase
keywords: Aspergilli; Cellulose; Endoglucanases; GH12; Glycoside hydrolases; Heterologous expression