Cloning, heterologous expression and biochemical characterization of a non-specific endoglucanase family 12 from Aspergillus terreus NIH2624
| Autor: | Gabriela L. Berto, Fernando Segato, Fabio M. Squina, Igor Polikarpov, Dyoni Matias de Oliveira, Flavio Henrique Moreira Souza, André Damasio, Mário T. Murakami, Bruno Vilas Boas Dias, Ana Paula Citadini |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Biophysics Gene Expression Cellulase 01 natural sciences Biochemistry Analytical Chemistry Fungal Proteins 03 medical and health sciences chemistry.chemical_compound Aspergillus nidulans 010608 biotechnology Glycoside hydrolase Aspergillus terreus Cloning Molecular Molecular Biology chemistry.chemical_classification biology CELULOSE biology.organism_classification Recombinant Proteins Enzyme assay Xyloglucan Aspergillus 030104 developmental biology Enzyme chemistry biology.protein Heterologous expression |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 1570-9639 |
| DOI: | 10.1016/j.bbapap.2017.01.003 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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