Improved heterologous protein production by a tripeptidyl peptidase gene (AosedD) disruptant of the filamentous fungus Aspergillus oryzae

Zhu, Lin; Nemoto, Toru; Yoon, Jaewoo; Maruyama, Jun‐ichi; Kitamoto, Katsuhiko

doi:10.2323/jgam.58.199

Cited by 5 publications

(1 citation statement)

References 26 publications

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“…To further solve the bottlenecks limiting the production of heterologous protein by A. oryzae, another tripeptidyl peptidase gene, AosedD, which may be involved in the proteolytic degradation of recombinant proteins, was selected to construct its deletion strain for the production of heterologous proteins. In the AosedD deletion mutant, the production levels of recombinant HLY and CHY improved approximately 1.7-and 2.9-fold, respectively, relative to their control strains (Zhu et al, 2012). The results show that AoSedD is one of the main proteases involved in the degradation of heterologous recombinant proteins in A. oryzae.…”

Section: Disruption Of Proteinase Genes In a Oryzaementioning

confidence: 83%

Advances in Genetic Engineering Technology and Its Application in the Industrial Fungus Aspergillus oryzae

Jin

Wang

et al. 2021

Front. Microbiol.

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The filamentous fungus Aspergillus oryzae is an important strain in the traditional fermentation and food processing industries and is often used in the production of soy sauce, soybean paste, and liquor-making. In addition, A. oryzae has a strong capacity to secrete large amounts of hydrolytic enzymes; therefore, it has also been used in the enzyme industry as a cell factory for the production of numerous native and heterologous enzymes. However, the production and secretion of foreign proteins by A. oryzae are often limited by numerous bottlenecks that occur during transcription, translation, protein folding, translocation, degradation, transport, secretion, etc. The existence of these problems makes it difficult to achieve the desired target in the production of foreign proteins by A. oryzae. In recent years, with the decipherment of the whole genome sequence, basic research and genetic engineering technologies related to the production and utilization of A. oryzae have been well developed, such as the improvement of homologous recombination efficiency, application of selectable marker genes, development of large chromosome deletion technology, utilization of hyphal fusion techniques, and application of CRISPR/Cas9 genome editing systems. The development and establishment of these genetic engineering technologies provided a great deal of technical support for the industrial production and application of A. oryzae. This paper reviews the advances in basic research and genetic engineering technologies of the fermentation strain A. oryzae mentioned above to open up more effective ways and research space for the breeding of A. oryzae production strains in the future.

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Section: Disruption Of Proteinase Genes In a Oryzaementioning

confidence: 83%

Advances in Genetic Engineering Technology and Its Application in the Industrial Fungus Aspergillus oryzae

Jin

Wang

et al. 2021

Front. Microbiol.

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Further enhanced production of heterologous proteins by double-gene disruption (ΔAosedD ΔAovps10) in a hyper-producing mutant of Aspergillus oryzae

Zhu

Maruyama

Kitamoto

2013

Appl Microbiol Biotechnol

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The filamentous fungus Aspergillus oryzae is used as one of the most favored hosts for heterologous protein production due to its ability to secrete large amounts of proteins into the culture medium. We previously generated a hyper-producing mutant strain of A. oryzae, AUT1, which produced 3.2- and 2.6-fold higher levels of bovine chymosin (CHY) and human lysozyme (HLY), respectively, compared with the wild-type strain. However, further enhancement of heterologous protein production by multiple gene disruption is difficult because of the low gene-targeting efficiency in strain AUT1. Here, we disrupted the ligD gene, which is involved in nonhomologous recombination, and the pyrG gene to create uridine/uracil auxotrophy in strain AUT1, to generate a hyper-producing mutant applicable to pyrG marker recycling with highly efficient gene targeting. We generated single and double disruptants of the tripeptidyl peptidase gene AosedD and vacuolar sorting receptor gene Aovps10 in the hyper-producing mutant background, and found that all disruptants showed significant increases in heterologous protein production. Particularly, double disruption of the Aovps10 and AosedD genes increased the production levels of CHY and HLY by 1.6- and 2.1-fold, respectively, compared with the parental strain. Thus, we successfully generated a fungal host for further enhancing the heterologous protein production ability by combining mutational and molecular breeding techniques.

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Reduction of Extracellular Proteases Increased Activity and Stability of Heterologous Protein in $${ Aspergillus}$$ A s p e r g i l l u s $${ niger}$$ n i g e r

et al. 2017

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Improved heterologous protein production by a tripeptidyl peptidase gene (AosedD) disruptant of the filamentous fungus Aspergillus oryzae

Cited by 5 publications

References 26 publications

Advances in Genetic Engineering Technology and Its Application in the Industrial Fungus Aspergillus oryzae

Advances in Genetic Engineering Technology and Its Application in the Industrial Fungus Aspergillus oryzae

Further enhanced production of heterologous proteins by double-gene disruption (ΔAosedD ΔAovps10) in a hyper-producing mutant of Aspergillus oryzae

Reduction of Extracellular Proteases Increased Activity and Stability of Heterologous Protein in $${ Aspergillus}$$ A s p e r g i l l u s $${ niger}$$ n i g e r

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