Expression of xylanase enzymes from thermophilic microorganisms in fungal hosts
Autor: | Maristela Azevedo, Angela Cziferszky, Fabricia Paula de Faria, Helena Nevalainen, Peter L. Bergquist, Moreland D. Gibbs, Valentino S Junior Te'o |
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Rok vydání: | 2002 |
Předmět: |
DNA
Bacterial Trichoderma Kluyveromyces lactis Bacteria Base Sequence Sequence Homology Amino Acid biology Thermophile Heterologous General Medicine biology.organism_classification Microbiology Recombinant Proteins Yeast Xylan Endo-1 3-beta-Xylosidase Kluyveromyces Xylosidases Xylanase Molecular Medicine Dictyoglomus thermophilum Electrophoresis Polyacrylamide Gel Amino Acid Sequence Trichoderma reesei |
Zdroj: | Extremophiles. 6:177-184 |
ISSN: | 1433-4909 1431-0651 |
DOI: | 10.1007/s00792-001-0252-5 |
Popis: | Bulk production of xylanases from thermophilic microorganisms is a prerequisite for their use in industrial processes. As effective secretors of gene products, fungal expression systems provide a promising, industrially relevant alternative to bacteria for heterologous enzyme production. We are currently developing the yeast Kluyveromyces lactis and the filamentous fungus Trichoderma reesei for the extracellular production of thermophilic enzymes for the pulp and paper industry. The K. lactis system has been tested with two thermophilic xylanases and secretes gram amounts of largely pure xylanase A from Dictyoglomus thermophilum in chemostat culture. The T. reesei expression system involves the use of the cellobiohydrolase I (CBHI) promoter and gene fusions for the secretion of heterologous thermostable xylanases of both bacterial and fungal origin. We have reconstructed the AT-rich xynB gene of Dictyoglomus thermophilum according to Trichoderma codon preferences and demonstrated a dramatic increase in expression. A heterologous fungal gene, Humicola grisea xyn2, could be expressed without codon modification. Initial amounts of the XYN2 protein were of a gram per liter range in shake-flask cultivations, and the gene product was correctly processed by the heterologous host. Comparison of the expression of three thermophilic heterologous microbial xylanases in T. reesei demonstrates the need for addressing each case individually. |
Databáze: | OpenAIRE |
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