A fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae
article
Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-staterelated physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilmreactor for the production of a Gla::green fluorescent protein (GFP) fusion protein by Aspergillus oryzae. The biofilmreactor 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 uniformcolonization of the packing by the fungal biomass and leads to a better quality of the fusion protein.
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 uniformcolonization of the packing by the fungal biomass and leads to a better quality of the fusion protein.
Topics
Fungal biofilm
Bioreactor
Scale-up
Recombinant protein
Antigen-antibody reactions
Aspergillus
Biofilms
Biomass
Bioreactors
Electrophoresis
Enzyme activity
Fermentation
Fungi
Metabolites
Physiology
Recombinant proteins
Shear stress
Comparative analysis
Extracellular medium
Green fluorescent protein
Physiological mechanisms
Secondary metabolites
Solid-state fermentation
Submerged fermentation
Proteins
Amylase
Glucan 1,4 alpha glucosidase
Glucoamylase green fluorescent protein fusion protein
Green fluorescent protein
Hybrid protein
Unclassified drug
Fermentation
Fungus
Metal
Physiological response
Protein
Qualitative analysis
Recirculating system
Recombination
Secondary metabolite
Alkalinization
Amino acid sequence
Aspergillus niger
Aspergillus oryzae
Autolysis
Biofilm reactor
Biotransformation
Carbon source
Controlled study
Enzyme activity
Extracellular space
Fungal biomass
Fungal colonization
Fungus culture
Fungus growth
Glucose intake
Hydrolysis
Immersion
Immunodetection
Nonhuman
PH measurement
Protein analysis
Shear stress
Solid state fermentation
Submerged fermentation
Western blotting
TNO Identifier
525956
DOI
https://dx.doi.org/10.1007/s00253-015-6608-z
Source
Applied Microbiology and Biotechnology, 99(15), pp. 6241-6254.
Pages
6241-6254
Files
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