High-level heterologous protein production using an attenuated selection marker in &lt;i&gt;Aspergillus sojae&lt;/i&gt;

Araki, Yasuko; Yuzuki, Masanobu; Masakari, Yosuke; Sato, Atsushi; Gomi, Keiko; Hara, Seiichi

doi:10.2323/jgam.2020.05.002

Cited by 2 publications

(6 citation statements)

References 22 publications

(21 reference statements)

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“…A. sojae KP-del (Araki et al, 2020) was used as a host strain for heterologous expression of each asc gene (DDBJ/ ENA/GenBank accession LC406756 and LC406757). All expression cassettes of asc genes were composed of a tef promoter (Ptef ) and an alp terminator (Talp), derived from A. sojae (Araki et al, 2020), and codon-optimized synthetic asc genes, without introns.…”

Section: Strainsmentioning

confidence: 99%

“…sojae KP-del (Araki et al, 2020) was used as a host strain for heterologous expression of each asc gene (DDBJ/ ENA/GenBank accession LC406756 and LC406757). All expression cassettes of asc genes were composed of a tef promoter (Ptef ) and an alp terminator (Talp), derived from A. sojae (Araki et al, 2020), and codon-optimized synthetic asc genes, without introns. An expression cassette of cprA was composed of a gpd promoter (Pgpd), a cprA homolog, and an amy terminator (Tamy), and amplified from the A. sojae NBRC4239 genome DNA using the following primers: P-gpd-F,R for Pgpd, AscprA-F, R for cprA homolog, and T-amy-F, R for Tamy.…”

Section: Strainsmentioning

confidence: 99%

“…The loop-out region for pyrG marker recycling was used, similar to the case of egtA integration (Hara and Ichikawa, 2019). The DNA fragment for the integration of each ascA-E, I and J gene was composed of a 5′-arm, Ptef, the asc gene, Talp, a loop-out region, a pyrG marker (Araki et al, 2020), and a 3′-arm, in that linear order. The DNA fragment for co-integration of ascH and cprA was composed of a 5′-arm, Ptef, ascH, Talp, a pyrG marker, Pgpd, cprA, Tamy, and a 3′-arm, in that linear order.…”

Section: Strainsmentioning

confidence: 99%

“…The multi-copy integrative vector containing the attenuated pyrG marker (pyrG56) was used to obtain As-CAB_ mcD and As-mcCABD (Araki et al, 2020). For construction of As-CAB_mcD, a pyrG disruptant of As-CAB was transformed using the pUC19-pG56-ascD vector, containing the ascD expression cassette.…”

Section: Strainsmentioning

confidence: 99%

“…For construction of As-CAB_mcD, a pyrG disruptant of As-CAB was transformed using the pUC19-pG56-ascD vector, containing the ascD expression cassette. For construction of As-mcCABD, P6-1 (Araki et al, 2020) was simultaneously transformed using two vectors, namely pUC19-pG56-as-cC+A and pUC19-pG56-ascB+D, containing expression cassettes of ascC and ascA, or ascB and ascD.…”

Section: Strainsmentioning

confidence: 99%

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Heterologous production of ascofuranone and ilicicolin A in <i>Aspergillus sojae</i>

Araki

Shinohara

Hara

et al. 2022

J. Gen. Appl. Microbiol.

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Ascofuranone and its precursor, ilicicolin A, are secondary metabolites with various pharmacological activities that are produced by Acremonium egyptiacum. In particular, ascofuranone strongly inhibits trypanosome alternative oxidase and represents a potential drug candidate against African trypanosomiasis. However, difficulties associated with industrial production of ascofuranone by A. egyptiacum, specifically the co-production of ascochlorin, which inhibits mammalian respiratory chain complex III at low concentrations, has precluded its widespread application. Therefore, in this study, ascofuranone biosynthetic genes (ascA-E and H-J) were heterologously expressed in Aspergillus sojae, which produced very low-levels of endogenous secondary metabolites under conventional culture conditions. As a result, although we obtained transformants producing both ilicicolin A and ascofuranone, they were produced only when an adequate concentration of chloride ions was added to the medium. In addition, we succeeded in increasing the production of ilicicolin A, by enhancing the expression of the rate-determining enzyme AscD, using a multi-copy integration system. The heterologous expression approach described here afforded the production of both ascofuranone and ilicicolin A, allowing for their development as therapeutics.

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Section: Strainsmentioning

confidence: 99%

Section: Strainsmentioning

confidence: 99%

Section: Strainsmentioning

confidence: 99%

Section: Strainsmentioning

confidence: 99%

Section: Strainsmentioning

confidence: 99%

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