Improved recombinant protein production in Aspergillus oryzae lacking both α-1,3-glucan and galactosaminogalactan in batch culture with a lab-scale bioreactor
| Autor: | Keietsu Abe, Ryutaro Orita, Shunya Susukida, Ken Miyazawa, Masahiro Hitosugi, Yoshikazu Kato, Yuka Kamachi, Akira Yoshimi, Ayumu Takeuchi, Takahiro Shintani, Silai Zhang, Kiyoaki Muto, Keisuke Komeiji, Hikaru Ichikawa |
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| Rok vydání: | 2022 |
| Předmět: |
Hypha
biology Chemistry Aspergillus oryzae Mutant Wild type Bioengineering biology.organism_classification Applied Microbiology and Biotechnology Recombinant Proteins law.invention Microbiology Cell wall Bioreactors Batch Cell Culture Techniques Polysaccharides law Bioreactor Recombinant DNA Glucans Mycelium Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 133:39-45 |
| ISSN: | 1389-1723 |
| DOI: | 10.1016/j.jbiosc.2021.09.010 |
| Popis: | Filamentous fungi are used as production hosts for various commercially valuable enzymes and chemicals including organic acids and secondary metabolites. We previously revealed that α-1,3-glucan and galactosaminogalactan (GAG) contribute to hyphal aggregation in the industrial fungus Aspergillus oryzae, and that production of recombinant protein in shake-flask culture is higher in a mutant lacking both α-1,3-glucan and GAG (AGΔ-GAGΔ) than in the parental strain. Here, we compared the productivity of the wild type, AGΔ-GAGΔ, and mutants lacking α-1,3-glucan (AGΔ) or GAG (GAGΔ) in batch culture with intermittent addition of glucose in a 5-L lab-scale bioreactor. The hyphae of the wild type and all mutants were dispersed by agitation, although the wild type and AGΔ formed small amounts of aggregates. Although mycelial weight was similar among the strains, the concentration of a secreted recombinant protein (CutL1) was the highest in AGΔ-GAGΔ. Evaluation of fluid properties revealed that the apparent viscosities of mycelial cultures of the wild type and AGΔ-GAGΔ decreased as the agitation speed was increased. The apparent viscosity of the AGΔ-GAGΔ culture tended to be lower than that of the wild-type strain at each agitation speed, and was significantly lower at 600 rpm. Overall, the lack of α-1,3-glucan and GAG in the hyphae improved culture rheology, resulting in an increase in recombinant protein production in AGΔ-GAGΔ. This is the first report of flow behavior improvement by a cell-surface component defect in a filamentous fungus. |
| Databáze: | OpenAIRE |
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