High-yield secretion of multiple client proteins in Aspergillus
| Autor: | Stephen R. Decker, André Damasio, Fabio M. Squina, Fernando Segato, Thiago Augusto Gonçalves, Rolf A. Prade, Rosymar Coutinho de Lucas |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Genetic Markers
Genetic Vectors Molecular Sequence Data Bioengineering Heterologous expression and secretion of multiple proteins Protein Engineering Applied Microbiology and Biotechnology Biochemistry Aspergillus nidulans Extracellular Protein biosynthesis Protein production Secretion Gene DNA Primers chemistry.chemical_classification biology Base Sequence Protein Stability Fungi Protein engineering Hydrogen-Ion Concentration biology.organism_classification Recombinant Proteins Enzyme Secretory protein Aspergillus chemistry Protein secretion Biotechnology Plasmids |
| Zdroj: | LOCUS Repositório Institucional da UFV Universidade Federal de Viçosa (UFV) instacron:UFV |
| Popis: | Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished. |
| Databáze: | OpenAIRE |
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