Details

Autor(en) / Beteiligte

• Hollands, Kerry
• Baron, Christopher M.
• Gibson, Katharine J.
• Kelly, Kristen J.
• Krasley, Elizabeth A.
• Laffend, Lisa A.
• Lauchli, Ryan M.
• Maggio-Hall, Lori A.
• Nelson, Mark J.
• Prasad, Jahnavi C.
• Ren, Yixin
• Rice, Barbara A.
• Rice, Gregory H.
• Rothman, Steven C.

Titel

Engineering two species of yeast as cell factories for 2′-fucosyllactose

Ist Teil von

• Metabolic engineering, 2019-03, Vol.52, p.232-242

Ort / Verlag

Belgium: Elsevier Inc

Erscheinungsjahr

2019

Quelle

MEDLINE

Beschreibungen/Notizen

• Oligosaccharides present in human breast milk have been linked to beneficial effects on infant health. Inclusion of these human milk oligosaccharides (HMOs) in infant formula can recapitulate these health benefits. As a result, there is substantial commercial interest in a cost-effective source of HMOs as infant formula ingredients. Here we demonstrate that the yeast species Saccharomyces cerevisiae and Yarrowia lipolytica both can be engineered to produce 2′-fucosyllactose (2′FL), which is the most abundant oligosaccharide in human breast milk, at high titer and productivity. Both yeast species were modified to enable uptake of lactose and synthesis of GDP-fucose – the two precursors of 2′FL – by installing a lactose transporter and enzymes that convert GDP-mannose to GDP-fucose. Production of 2′FL was then enabled by expression of α-1,2-fucosyltransferases from various organisms. By screening candidate transporters from a variety of sources, we identified transporters capable of exporting 2′FL from yeast, which is a key consideration for any biocatalyst for 2′FL production. In particular, we identified CDT2 from Neurospora crassa as a promising target for further engineering to improve 2′FL efflux. Finally, we demonstrated production of 2′FL in fermenters at rates and titers that indicate the potential of engineered S. cerevisiae and Y. lipolytica strains for commercial 2′FL production. •Engineered two distantly related species of yeast to produce 2′-fucosyllactose.•Fermentation rates and titers indicate potential of yeast for commercial production.•Identified transporters that enable yeast to export 2′-fucosyllactose.•N. crassa CDT2 shows promise for engineering to enhance 2′-fucosyllactose efflux.