Direct Liquid-Culture Screening for Evaluating the Production of Heterologous Proteins Using an Auxotrophic Mutant of Aspergillus Oryzae

10.3791/61219 ◽  
2020 ◽  
Author(s):  
Satoru Shinkawa ◽  
Shigenobu Mitsuzawa
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Auxotrophic Mutant ◽  
Heterologous Proteins

Characteristics of α-glucosidase production from recombinant Aspergillus oryzae by membrane-surface liquid culture in comparison with various cultivation methods

2004 ◽  
Vol 98 (3) ◽  
pp. 200-206 ◽  
Author(s):  
Masakazu Morita ◽  
Hiroko Shimamura ◽  
Natsuko Ishida ◽  
Koreyoshi Imamura ◽  
Takaharu Sakiyama ◽  
...  
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Membrane Surface ◽  
Surface Liquid ◽  
Cultivation Methods

scholarly journals Both galactosaminogalactan and α-1,3-glucan contribute to aggregation of Aspergillus oryzae hyphae in liquid culture

2019 ◽  
Author(s):  
Ken Miyazawa ◽  
Akira Yoshimi ◽  
Motoaki Sano ◽  
Fuka Tabata ◽  
Asumi Sugahara ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Aspergillus Oryzae ◽  
Double Mutant ◽  
Liquid Culture ◽  
Bacterial Cells ◽  
Wild Type ◽  
Hydrogen Bond Formation ◽  
Relative Contribution ◽  
In The Wild ◽  
Increase Productivity

AbstractFilamentous fungi generally form aggregated hyphal pellets in liquid culture. We previously reported that α-1,3-glucan-deficient mutants of Aspergillus nidulans did not form hyphal pellets and their hyphae were fully dispersed, and we suggested that α-1,3-glucan functions in hyphal aggregation. Yet, Aspergillus oryzae α-1,3-glucan-deficient (AGΔ) mutants still form small pellets; therefore, we hypothesized that another factor responsible for forming hyphal pellets remains in these mutants. Here, we identified an extracellular matrix polysaccharide galactosaminogalactan (GAG) as such a factor. To produce a double mutant of A. oryzae (AG-GAGΔ), we disrupted the genes required for GAG biosynthesis in an AGΔ mutant. Hyphae of the double mutant were fully dispersed in liquid culture, suggesting that GAG is involved in hyphal aggregation in A. oryzae. Addition of partially purified GAG fraction to the hyphae of the AG-GAGΔ strain resulted in formation of mycelial pellets. Acetylation of the amino group in galactosamine of GAG weakened GAG aggregation, suggesting that hydrogen bond formation by this group is important for aggregation. Genome sequences suggest that α-1,3-glucan, GAG, or both are present in many filamentous fungi and thus may function in hyphal aggregation in these fungi. We also demonstrated that production of a recombinant polyesterase, CutL1, was higher in the AG-GAGΔ strain than in the wild-type and AGΔ strains. Thus, controlling hyphal aggregation factors of filamentous fungi may increase productivity in the fermentation industry.ImportanceProduction using filamentous fungi is an important part of the fermentation industry, but hyphal aggregation in these fungi in liquid culture limits productivity compared with that of yeast or bacterial cells. We found that galactosaminogalactan and α-1,3-glucan both function in hyphal aggregation in Aspergillus oryzae, and that the hyphae of a double mutant deficient in both polysaccharides become fully dispersed in liquid culture. We also revealed the relative contribution of α-1,3-glucan and galactosaminogalactan to hyphal aggregation. Recombinant protein production was higher in the double mutant than in the wild-type strain. Our research provides a potential technical innovation for the fermentation industry that uses filamentous fungi, as regulation of the growth characteristics of A. oryzae in liquid culture may increase productivity.

Development of a cylindrical apparatus for membrane-surface liquid culture and production of kojic acid using Aspergillus oryzae NRRL484

1998 ◽  
Vol 85 (5) ◽  
pp. 488-494 ◽  
Author(s):  
Yasushi Wakisaka ◽  
Takayuki Segawa ◽  
Koreyoshi Imamura ◽  
Takaharu Sakiyama ◽  
Kazuhiro Nakanishi
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Kojic Acid ◽  
Membrane Surface ◽  
Surface Liquid

Production of kojic acid by membrane-surface liquid culture of Aspergillus oryzae NRRL484

1995 ◽  
Vol 80 (1) ◽  
pp. 41-45 ◽  
Author(s):  
Akinori Ogawa ◽  
Yasushi Wakisaka ◽  
Takaaki Tanaka ◽  
Takaharu Sakiyama ◽  
Kazuhiro Nakanishi
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Kojic Acid ◽  
Membrane Surface ◽  
Surface Liquid

Identification of genes from Aspergillus oryzae that are preferentially expressed in membrane-surface liquid culture

2006 ◽  
Vol 102 (5) ◽  
pp. 470-473 ◽  
Author(s):  
Bin Feng ◽  
Masakazu Morita ◽  
Hiroyuki Imanaka ◽  
Koreyoshi Imamura ◽  
Kazuhiro Nakanishi
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Membrane Surface ◽  
Surface Liquid

scholarly journals Increased enzyme production under liquid culture conditions in the industrial fungus Aspergillus oryzae by disruption of the genes encoding cell wall α-1,3-glucan synthase

2016 ◽  
Vol 80 (9) ◽  
pp. 1853-1863 ◽  
Author(s):  
Ken Miyazawa ◽  
Akira Yoshimi ◽  
Silai Zhang ◽  
Motoaki Sano ◽  
Mayumi Nakayama ◽  
...  
Keyword(s):  
Cell Wall ◽  
Aspergillus Oryzae ◽  
Enzyme Production ◽  
Liquid Culture ◽  
Culture Conditions ◽  
Glucan Synthase ◽  
Genes Encoding

Production of neutral protease by membrane-surface liquid culture of Aspergillus oryzae IAM2704

1995 ◽  
Vol 80 (1) ◽  
pp. 35-40 ◽  
Author(s):  
Akinori Ogawa ◽  
Akinori Yasuhara ◽  
Takaaki Tanaka ◽  
Takaharu Sakiyama ◽  
Kazuhiro Nakanishi
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture ◽  
Membrane Surface ◽  
Neutral Protease ◽  
Surface Liquid

scholarly journals Both Galactosaminogalactan and α-1,3-Glucan Contribute to Aggregation of Aspergillus oryzae Hyphae in Liquid Culture

2019 ◽  
Vol 10 ◽  
Author(s):  
Ken Miyazawa ◽  
Akira Yoshimi ◽  
Motoaki Sano ◽  
Fuka Tabata ◽  
Asumi Sugahara ◽  
...  
Keyword(s):  
Aspergillus Oryzae ◽  
Liquid Culture

Strategies for Increasing the Production Level of Heterologous Proteins in Aspergillus oryzae

2014 ◽  
pp. 149-164 ◽  
Author(s):  
Mizuki Tanaka ◽  
Katsuya Gomi
Keyword(s):  
Aspergillus Oryzae ◽  
Production Level ◽  
Heterologous Proteins
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