Microorganisms offer a tremendous potential as cell factories, and they are indeed been used by humans since the previous centuries for biotransformations. Among them, yeasts combine the advantage of a unicellular state with a eukaryotic organization. Moreover, in the era of biorefineries, their biodiversity can offer solutions to specific process constraints. Zygosaccharomyces bailii, an ascomycete budding yeast, is widely known for its peculiar tolerance to different stresses, among which are organic acids. Moreover, the recent reclassification of the species, including diverse hybrids, is further expanding both fundamental and applied interests. It is therefore reasonable that despite the possibility to apply with this yeast some of the molecular tools and protocols routinely used to manipulate Saccharomyces cerevisiae, adjustments and optimizations are necessary. Here we describe in detail the methods for determining chromosome number, size, and aneuploidy, transformation, classical target gene disruption or gene integration, and designing of episomal expression plasmids helpful for engineering the yeast Z. bailii .

Branduardi, P., Barroso, L., Dato, L., Louis, E., Porro, D. (2022). Molecular Tools for Leveraging the Potential of the Acid-Tolerant Yeast Zygosaccharomyces bailii as Cell Factory. In V. Mapelli, M. Bettiga (a cura di), Yeast Metabolic Engineering : Methods and Protocols (pp. 179-204). Humana Press Inc. [10.1007/978-1-0716-2399-2_11].

Molecular Tools for Leveraging the Potential of the Acid-Tolerant Yeast Zygosaccharomyces bailii as Cell Factory

Branduardi, P
;
Dato, L;Porro, D
2022

Abstract

Microorganisms offer a tremendous potential as cell factories, and they are indeed been used by humans since the previous centuries for biotransformations. Among them, yeasts combine the advantage of a unicellular state with a eukaryotic organization. Moreover, in the era of biorefineries, their biodiversity can offer solutions to specific process constraints. Zygosaccharomyces bailii, an ascomycete budding yeast, is widely known for its peculiar tolerance to different stresses, among which are organic acids. Moreover, the recent reclassification of the species, including diverse hybrids, is further expanding both fundamental and applied interests. It is therefore reasonable that despite the possibility to apply with this yeast some of the molecular tools and protocols routinely used to manipulate Saccharomyces cerevisiae, adjustments and optimizations are necessary. Here we describe in detail the methods for determining chromosome number, size, and aneuploidy, transformation, classical target gene disruption or gene integration, and designing of episomal expression plasmids helpful for engineering the yeast Z. bailii .
Capitolo o saggio
Contour-clamped homogeneous electric field (CHEF) gel electrophoresis; Plasmids; Promoters; Targeted gene deletion; Yeast transformation; Zygosaccharomyces bailii; Acids; Humans; Saccharomyces cerevisiae; Saccharomycetales; Zygosaccharomyces
English
Yeast Metabolic Engineering : Methods and Protocols
978-1-0716-2398-5
Branduardi, P., Barroso, L., Dato, L., Louis, E., Porro, D. (2022). Molecular Tools for Leveraging the Potential of the Acid-Tolerant Yeast Zygosaccharomyces bailii as Cell Factory. In V. Mapelli, M. Bettiga (a cura di), Yeast Metabolic Engineering : Methods and Protocols (pp. 179-204). Humana Press Inc. [10.1007/978-1-0716-2399-2_11].
Branduardi, P; Barroso, L; Dato, L; Louis, E; Porro, D
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/397342
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