Adaptive laboratory evolution of Yarrowia lipolytica improves ferulic acid tolerance

Rapid Enabling of Gluconobacter oxydans Resistance to High D-Sorbitol Concentration and High Temperature by Microdroplet-Aided Adaptive Evolution

“…The substrate/product resistance of microorganisms is vital for their applications in industrial fermentation processes (Mohamed et al, 2020;Wang et al, 2021). In recent years, many reports on substrate/product tolerance of microbial strains, such as Clostridium acetobutylicum and Escherichia coli (Alsaker et al, 2010;Zhang et al, 2015), have been published, but studies on the tolerance of G. oxydans are few.…”

Yarrowia lipolytica strains and their biotechnological applications: how natural biodiversity and metabolic engineering could contribute to cell factories improvement

“…However, the erythritol yield was considerably lower in this selected strain, which prompted the use of genetic engineering as a last resort: the authors performed a transcriptome analysis on their thermotolerant strain in order to identify the genes linked to this phenotype, reconstructed the thermotolerant phenotype using a surrogate Po1f strain and finally transferred the target gene modifications into a BBE-17Ku70URA3 strain. This GM strain was able to produce a three-fold higher erythritol yield, without detrimental effects on cell growth, at a temperature of 33 °C compared to the parent BBE-17T strain [271]. Thus, such an example illustrates the limits of traditional mutagenesis, even when combined to adaptative evolution strategies, when used alone as an alternative to genetic engineering.…”

“…It appeared that single cell FACS was favoring intracellular riboflavin accumulation when droplet FACS was favoring extracellular product accumulation [266]. Beside this example, adaptative laboratory evolution strategies have recently been applied in Y. lipolytica for various purposes: restoring the glucose metabolism of a strain engineered for succinic acid production [267]; selecting the ionic liquid-tolerant YlCW001 strain [137]; improving limonene tolerance during its production by GM strains [268]; enhancing thermotolerance during industrial fermentation for erythritol production [269]; increasing tolerance to aromatic aldehydes or to ferulic acid, both for a more efficient lignocellulose valorization [270,271].…”

“…Interestingly, some genetic engineering technics were implemented in their project, but not for the design of the production strain: these tools were used to build a genetically-encoded erythritol biosensor strain (using an erythritol-responsive transcription factor to activate expression of an eGFP-encoding gene, leading to a fluorescence signal) that was used in the high-throughput screening test applied to the mutant strains generated through a combined UV/ARTP mutagenesis step [288]. The same authors pursued the optimization of their erythritol production bioprocess by applying an adaptative evolution strategy to the producing strain BBE-17 in order to enhance its thermotolerance during industrial fermentation, as evoked above in Section 4.5.3 [271]. Starting with a strain with an optimum temperature of 30 °C, they applied a progressive adaptive evolution scheme that allowed them to obtain, after 11 months of continuous cultivation and selection, an improved strain with an optimum temperature of 35 °C.…”

Yarrowia lipolytica Strains and Their Biotechnological Applications: How Natural Biodiversity and Metabolic Engineering Could Contribute to Cell Factories Improvement

“…It appeared that single cell FACS was favoring intracellular riboflavin accumulation when droplet FACS was favoring extracellular product accumulation [305]. In addition to this example, adaptative laboratory evolution strategies have recently been applied in Y. lipolytica for various purposes: restoring the glucose metabolism of a Po1f derivative engineered for succinic acid production [306]; enhancing lipid storage from ACA-DC 50109 strain [135]; selecting the ionic liquid-tolerant YlCW001 strain [161]; improving limonene tolerance during its production by GM strains [307]; enhancing thermotolerance during industrial fermentation for erythritol production [308]; increasing tolerance to aromatic aldehydes or to ferulic acid, both for a more efficient lignocellulose valorization [309,310].…”