A fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae
| Autor: | Quentin Zune, S. Gofflot, Frank Delvigne, Jean-Claude Twizere, Frédéric Francis, Anissa Delepierre, Peter J. Punt, J. Bauwens, Thomas Bawin, Dominique Toye |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Autolysis (biology)
Unclassified drug Recirculating system Physiology Scale-up Aspergillus oryzae Hybrid protein Gene Expression Biofilm reactor Applied Microbiology and Biotechnology Western blotting Green fluorescent protein Bioreactors Physiological response Life Solid-state fermentation Immersion Metabolites Alkalinization Glucoamylase green fluorescent protein fusion protein Protein analysis Extracellular space Biomass Enzyme activity Physiological mechanisms Promoter Regions Genetic Biotransformation Recombinant proteins Immunodetection Fungus Fungus culture Metal Hydrolysis Comparative analysis General Medicine Extracellular medium Submerged fermentation Aspergillus Biochemistry Aspergillus niger Healthy Living Biotechnology Electrophoresis Recombinant protein Recombinant Fusion Proteins Antigen-antibody reactions Green Fluorescent Proteins Bioreactor Biology Secondary metabolite Amino acid sequence Fungal biomass Food and Nutrition Nutrition Shear stress Glucan 1 4 alpha glucosidase Secondary metabolites Protein Glucose intake Fungi Biofilm Proteins Amylase Nonhuman biology.organism_classification Fusion protein Fungal biofilm Recombination Solid state fermentation MSB - Microbiology and Systems Biology Biofilms Fermentation Carbon source ELSS - Earth Life and Social Sciences Glucan 1 4-alpha-Glucosidase Autolysis Qualitative analysis Controlled study Fungal colonization Fungus growth PH measurement |
| Zdroj: | Applied Microbiology and Biotechnology, 15, 99, 6241-6254 |
| ISSN: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-015-6608-z |
| Popis: | Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state-related physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilm reactor for the production of a Gla::green fluorescent protein (GFP) fusion protein by Aspergillus oryzae. The biofilm reactor comprises a metal structured packing allowing the attachment of the fungal biomass. Since the production of the target protein is under the control of the promoter glaB, specifically induced in solid-state fermentation, the biofilm mode of culture is expected to enhance the global productivity. Although production of the target protein was enhanced by using the biofilm mode of culture, we also found that fusion protein production is also significant when the submerged mode of culture is used. This result is related to high shear stress leading to biomass autolysis and leakage of intracellular fusion protein into the extracellular medium. Moreover, 2-D gel electrophoresis highlights the preservation of fusion protein integrity produced in biofilm conditions. Two fungal biofilm reactor designs were then investigated further, i.e. with full immersion of the packing or with medium recirculation on the packing, and the scale-up potentialities were evaluated. In this context, it has been shown that full immersion of the metal packing in the liquid medium during cultivation allows for a uniform colonization of the packing by the fungal biomass and leads to a better quality of the fusion protein. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|