Biosynthesis of compounds in yeast
AssigneeAmyris, Inc.
InventorsJessica Walter, Joshua A. Lerman, Michael Leavell, Benjamin Yap
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Abstract
Provided herein are genetically modified yeast cells capable of producing one or more human milk oligosaccharides. The yeast cells include one or more heterologous nucleic acids that encode enzymes of a human milk oligosaccharide biosynthetic pathway. The yeast cells do not include a heterologous nucleic acid encoding a fucokinase. Also provided are fermentation compositions including the disclosed genetically modified yeast cells, and related methods of producing and recovering human milk oligosaccharides generated by the yeast cells.
Claims28
A genetically modified yeast cell capable of producing one or more human milk oligosaccharides, the yeast cell comprising one or more heterologous nucleic acids, each nucleic acid independently encoding at least one enzyme of a human milk oligosaccharide biosynthetic pathway, wherein the yeast cell does not comprise a heterologous nucleic acid encoding a fucokinase, and wherein the one or more heterologous nucleic acids are integrated into the genome of the yeast cell.
A method of producing one or more human milk oligosaccharides, the method comprising:
- providing a population of genetically modified yeast cells capable of producing one or more human milk oligosaccharides, each yeast cell comprising one or more heterologous nucleic acids, each nucleic acid independently encoding at least one enzyme of a human milk oligosaccharide biosynthetic pathway, wherein the one or more heterologous nucleic acids are integrated into the genome of the yeast cell;
- feeding to the population a culture medium comprising sucrose and lactose, wherein the mass ratio of the sucrose to the lactose is less than 40;
- and
- culturing the yeast cells in the culture medium under conditions suitable for the yeast cells to produce the one or more human milk oligosaccharides.
A method of treating a fermentation composition, the method comprising:
- providing a fermentation composition comprising difucosyllactose;
- and
- contacting the fermentation composition with α1-3,4 fucosidase under conditions suitable for catalyzing the conversion of at least a portion of the difucosyllactose to 2′-fucosyllactose with the α1-3,4 fucosidase;
- wherein the fermentation composition comprises a population of genetically modified yeast cells capable of producing one or more human milk oligosaccharides, each yeast cell comprising one or more heterologous nucleic acids, each nucleic acid independently encoding at least one enzyme of a human milk oligosaccharide biosynthetic pathway, and wherein the genetically modified yeast cells do not comprise a heterologous nucleic acid encoding a fucokinase.
Dependencies are inferred from the claim text.