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Producing Biochemicals in Yarrowia Lipolytica from Xylose Through a Strain Mating Approach.

Efficient xylose conversion is an essential feature for producing renewable biochemical from lignocellulosic hydrolysate. However, rewiring xylose catabolism is challenging, especially for unconventional yeasts and previously engineered background strains. In this study, we demonstrate the efficacy of a yeast mating approach with Y. lipolytica that can combine a previously engineering and evolved xylose phenotype with a metabolite overproduction phenotype. Specifically, we mated several engineered Y. lipolytica strains that produce α-linolenic acid, riboflavin and triacetic acid lactone with an engineered and adapted xylose utilizing strain to obtain three diploid strains that rapidly produce these molecules directly from xylose. Specifically, titers of 0.52 g/L α-linolenic acid (ALA), 96.6 mg/L riboflavin and 2.9 g/L triacetic acid lactone (TAL) respectively from were obtained from xylose in flask cultures, and 1.42 g/L ALA in bioreactor condition. This total production level was generally on par or higher than the parental strain cultivated on glucose, although specific productivities decreased as a result of improved overall cell growth by the diploid strains. In the case of ALA, this lipid content reached similar levels to that of flaxseed oil. This result showcases the first study using strain mating in Y. lipolytica for producing biomolecules from xylose, and thus demonstrates the utility of this approach as a routine tool for metabolic engineering. This article is protected by copyright. All rights reserved.