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US 11,913,046 B2Granted
Increasing export of 2'fucosyllactose from microbial cells through the expression of a heterologous nucleic acid
AssigneeInbiose N.V.
InventorsKerry Hollands, Lori Anne Maggio-Hall, Steven Cary Rothman
Life of this patent
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Abstract
Microbial cells genetically engineered with a heterologous nucleic acid sequence that increases export of 2′ fucosyllactose are disclosed. Methods of increasing export of 2′ fucosyllactose from a microbial cell and for identifying a heterologous nucleic acid sequence that increases export of 2′ fucosyllactose from a microbial cell are also disclosed.
Claims17
A genetically engineered yeast cell, wherein:
- the cell comprises:
- a) at least one heterologous nucleic acid molecule encoding a transporter protein that facilitates export of 2′ fucosyllactose from the yeast cell, wherein the transporter protein is selected from the group consisting of
- a Sugars Will Eventually be Exported Transporter (SWEET) family transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 93, 94, 95, and 96,
- a Sugar Efflux Transporter A (SetA) family transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 88, 105, 106, 107, and 108, and
- a sugar porter family transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 65 and SEQ ID NO:
- 66;
- b) at least one heterologous nucleic acid molecule encoding a guanosine diphosphate (GDP)-mannose-4,6-dehydratase (EC 4.2.1.47);
- c) at least one heterologous nucleic acid molecule encoding a GDP-4-keto-6-D-deoxymannose epimerase-reductase (EC 1.1.1.271);
- and
- d) at least one heterologous nucleic acid molecule encoding a 2-α-L-fucosyltransferase (EC 2.4.1.69);
- and
- said yeast cell produces 2′ fucosyllactose.
12 more elements
A genetically engineered yeast cell that produces 2′ fucosyllactose, wherein the yeast cell comprises:
- a) at least one heterologous nucleic acid molecule encoding a transporter protein that facilitates export of 2′ fucosyllactose from the yeast cell;
- b) at least one heterologous nucleic acid molecule encoding a guanosine diphosphate (GDP)-mannose-4,6-dehydratase (EC 4.2.1.47);
- c) at least one heterologous nucleic acid molecule encoding a GDP-4-keto-6-D-deoxymannose epimerase-reductase (EC 1.1.1.271);
- d) at least one heterologous nucleic acid molecule encoding a 2-α-L-fucosyltransferase (EC 2.4.1.69);
- and
- e) at least one nucleic acid molecule encoding a lactose transporter having an amino acid sequence having 90% identity to peptide selected from the group consisting of SEQ ID NO:
- 24, 25, 26, 27 and 28.
4 more elements
A yeast cell, which is genetically engineered, the yeast cell comprising:
- a) at least one heterologous polynucleotide encoding a transporter protein that facilitates export of 2′ fucosyllactose from the yeast cell, wherein the transporter protein is selected from the group consisting of a Sugars Will Eventually be Exported Transporter (SWEET) family transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 93, 94, 95, and 96, a Sugar Efflux Transporter A (SetA) family transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 88, 105, 106, 107, and 108, and a Sugar porter family transporter having at least 90% identity to a peptide selected from SEQ ID NO:
- 65 and SEQ ID NO:
- 66;
- b) at least one heterologous polynucleotide encoding a guanosine diphosphate (GDP)-mannose-4,6-dehydratase (EC 4.2.1.47);
- c) at least one heterologous polynucleotide encoding a GDP-4-keto-6-D-deoxymannose epimerase-reductase (EC 1.1.1.271);
- and
- d) at least one heterologous polynucleotide encoding a 2-α-L-fucosyltransferase (EC 2.4.1.69);
- wherein the yeast cell produces 2′ fucosyllactose.
7 more elements
A yeast cell comprising:
- a) at least one heterologous polynucleotide encoding a transporter protein that facilitates export of 2′ fucosyllactose from the yeast cell, wherein the transporter protein is selected from the group consisting of a Sugars Will Eventually be Exported Transporter (SWEET) family transporter, a Sugar Efflux Transporter A (SetA) family transporter, and a sugar porter family transporter;
- b) at least one heterologous polynucleotide encoding a guanosine diphosphate (GDP)-mannose-4,6-dehydratase (EC 4.2.1.47);
- c) at least one heterologous polynucleotide encoding a GDP-4-keto-6-D-deoxymannose epimerase-reductase (EC 1.1.1.271);
- d) at least one heterologous polynucleotide encoding a 2-α-L-fucosyltransferase (EC 2.4.1.69);
- wherein the yeast cell produces 2′ fucosyllactose, and
- e) at least one polynucleotide encoding a lactose transporter having at least 90% identity to a peptide selected from the group consisting of SEQ ID NO:
- 24, 25, 26, 27, and 28.
4 more elements
Dependencies are inferred from the claim text.