New biotechnological applications for<i>Ashbya gossypii</i>: Challenges and perspectives

Aguiar, Tatiana Quinta; Silva, Rui; Domingues, Lucı́lia

doi:10.1080/21655979.2016.1234543

Cited by 18 publications

(11 citation statements)

References 30 publications

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“…The most well-known may be the proof of the whole genome duplication in S. cerevisiae [19,20]. The molecular analysis of riboflavin production and the novel bioengineering efforts to implement Ashbya as a platform strain for proteins, lipids or flavor compounds has made Ashbya a very interesting strain for the biotech industry as an alternative and addition to S. cerevisiae and other yeasts as well as for synthetic biology approaches [3,6,7,9,10,18]. Developmental biology contributed to the analysis of polarized hyphal growth, septation, nuclear migration and asynchronous nuclear divisions in a filamentous fungus [22][23][24]27,[44][45][46][47][48][49].…”

Section: Outlook: Open Research Questions and Unknown Territorymentioning

confidence: 99%

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Sporulation in Ashbya gossypii

Wendland

2020

JoF

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Ashbya gossypii is a filamentous ascomycete belonging to the yeast family of Saccharomycetaceae. At the end of its growth phase Ashbya generates abundant amounts of riboflavin and spores that form within sporangia derived from fragmented cellular compartments of hyphae. The length of spores differs within species of the genus. Needle-shaped Ashbya spores aggregate via terminal filaments. A. gossypii is a homothallic fungus which may possess a and α mating types. However, the solo-MATa type strain is self-fertile and sporulates abundantly apparently without the need of prior mating. The central components required for the regulation of sporulation, encoded by IME1, IME2, IME4, KAR4, are conserved with Saccharomyces cerevisiae. Nutrient depletion generates a strong positive signal for sporulation via the cAMP-PKA pathway and SOK2, which is also essential for sporulation. Strong inhibitors of sporulation besides mutations in the central regulatory genes are the addition of exogenous cAMP or the overexpression of the mating type gene MATα2. Sporulation has been dissected using gene-function analyses and global RNA-seq transcriptomics. This revealed a role of Msn2/4, another potential PKA-target, for spore wall formation and a key dual role of the protein A kinase Tpk2 at the onset of sporulation as well as for breaking the dormancy of spores to initiate germination. Recent work has provided an overview of ascus development, regulation of sporulation and spore maturation. This will be summarized in the current review with a focus on the central regulatory genes. Current research and open questions will also be discussed.

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Section: Outlook: Open Research Questions and Unknown Territorymentioning

confidence: 99%

“…Ashbya requires sucking insects to form plant infection as it does not produce penetration pegs or lytic enzymes. On the other hand, this makes Ashbya a suitable host for recombinant protein production [10].…”

Section: Introductionmentioning

confidence: 99%

Sporulation in Ashbya gossypii

Wendland

2020

JoF

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“…Indeed, the worldwide riboflavin market mostly relies on A. gossypii bioprocessing, which provides the vitamin as a safe additive for human foodstuffs and animal feeding (Schwechheimer et al ., ; Rychen et al ., ). In addition, A. gossypii has been also shown as an appropriate biotechnological chassis for other applications such as the production of folic acid (Serrano‐Amatriain et al ., ), microbial oils (Ledesma‐Amaro et al ., , ; Lozano‐Martínez et al ., ; Díaz‐Fernández et al ., ), nucleosides (Ledesma‐Amaro et al ., ) and recombinant proteins (Aguiar et al ., , ). The main advantages of A. gossypii as a microbial factory are based on the ability to grow using low‐cost substrates and an inexpensive downstream processing (Schwechheimer et al ., ).…”

Section: Introductionmentioning

confidence: 99%

One‐vector CRISPR/Cas9 genome engineering of the industrial fungus Ashbya gossypii

Jiménez

Muñoz‐Fernández

Ledesma‐Amaro

et al. 2019

Microbial Biotechnology

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Summary The filamentous fungus Ashbya gossypii is currently used for the industrial production of vitamin B2. Furthermore, the ability of A. gossypii to grow using low‐cost substrates together with the inexpensive downstream processing makes this fungus an attractive biotechnological chassis. Indeed, the production in A. gossypii of other high‐added value compounds such as folic acid, nucleosides and biolipids has been described. Hence, the development of new methods to expand the molecular toolkit for A. gossypii genomic manipulation constitutes an important issue for the biotechnology of this fungus. In this work, we present a one‐vector CRISPR/Cas9 system for genomic engineering of A. gossypii. We demonstrate the efficiency of the system as a marker‐less approach for nucleotide deletions and substitutions both with visible and invisible phenotypes. Particularly, the system has been validated for three types of genomic editions: gene inactivation, the genomic erasure of loxP scars and the introduction of point mutations. We anticipate that the use of the CRISPR/Cas9 system for A. gossypii will largely contribute to facilitate the genomic manipulations of this industrial fungus in a marker‐less manner.

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“…A. gossypii is a filamentous fungus of considerable industrial and biological significance, due to its capacity to overproduce riboflavin (vitamin B2) and to its unique genetic and biological features [9,10]. The genetic blockage of its pyrimidine pathway at the AgURA3 level (orotidine 5'-P to UMP) leads to increased riboflavin production on standard Ashbya Full Medium (AFM), a phenotype that is reverted when extra uridine/uracil (≥0.5/1 mM) is added to the medium [11].…”

Section: Introductionmentioning

confidence: 99%

Physiological characterization of a pyrimidine auxotroph exposes link between uracil phosphoribosyltransferase regulation and riboflavin production in Ashbya gossypii

et al. 2019

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The blockage of the de novo pyrimidine biosynthetic pathway at the orotidine-5′-phosphate decarboxylase level was previously demonstrated to affect riboflavin production in the industrial producer fungus Ashbya gossypii. However, the molecular basis for the unusual sensitivity to uracil displayed by the pyrimidine auxotroph A. gossypii Agura3 was unknown. Here, uridine was shown to be the only intermediate of the pyrimidine salvage pathway able to fully restore this mutant's growth. Conversely, uracil, which is routinely used to rescue pyrimidine auxotrophs, had a dose-dependent growth-inhibitory effect. Uracil phosphoribosyltransferase (UPRT) is the pyrimidine salvage pathway enzyme responsible for converting uracil to uridine monophosphate in the presence of phosphoribosyl pyrophosphate (PRPP). Characterization of the A. gossypii UPRT, as produced and purified from Escherichia coli, revealed that uracil concentrations above 1 mM negatively affected its activity, thus explaining the hypersensitivity of the Agura3 mutant to uracil. Accordingly, overexpression of the AgUPRT encoding-gene in A. gossypii Agura3 led to similar growth on rich medium containing 5 mM uracil or uridine. Decreased UPRT activity ultimately favors the preservation of PRPP, which otherwise may be directed to other pathways. In A. gossypii, increased PRPP availability promotes overproduction of riboflavin. Thus, this UPRT modulation mechanism reveals a putative means of saving precursors essential for riboflavin overproduction by this fungus. A similar uracil-mediated regulation mechanism of the UPRT activity is reported only in two protozoan parasites, whose survival depends on the availability of PRPP. Physiological evidence here discussed indicate that it may be extended to other distantly related flavinogenic fungi.

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New biotechnological applications forAshbya gossypii: Challenges and perspectives

Cited by 18 publications

References 30 publications

Sporulation in Ashbya gossypii

Sporulation in Ashbya gossypii

One‐vector CRISPR/Cas9 genome engineering of the industrial fungus Ashbya gossypii

Physiological characterization of a pyrimidine auxotroph exposes link between uracil phosphoribosyltransferase regulation and riboflavin production in Ashbya gossypii

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