The nonconventional yeast Debaryomyces hansenii is a promising cell factory. An extremophile oleaginous yeast in the CTG clade, D. hansenii has attractive catabolic, anabolic, and tolerance phenotypes. It naturally grows on the major monosaccharides of lignocellulosic biomass, specifically glucose, xylose, and L-arabinose. It produces the nutraceutical riboflavin and biofuel precursors like free fatty acids. It is osmotolerant and halotolerant, which could enable saltwater fermentations. Genetic transformation protocols have been developed for D. hansenii, but a dearth of genetic engineering parts and tools have limited its further development. Here, we report comprehensively “onboarding” D. hansenii into a platform nonconventional yeast host. First, we characterized its growth and antibiotic sensitivity. Then, using our accurate yeast genome assembly tool, we resequenced the genome along with other Debaryomyces strains of interest. We then improved transformation, designed standardized genomic integration, and established modular cloning for D. hansenii. Finally, we validated the function of genetic parts, including reporter genes, 24 promoters, and 24 terminators. This “onboarding” work provides the foundation for pathway engineering in this promising host, potentially enabling cell factories for nutraceuticals and biofuels.

Saltwater cell factories enabled by synthetic biology in the yeast Debaryomyces hansenii

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.