US 11,913,046 B2
Increasing export of 2'fucosyllactose from microbial cells through the expression of a heterologous nucleic acid
Kerry Hollands, Newark, DE (US); Lori Anne Maggio-Hall, Wilmington, DE (US); and Steven Cary Rothman, Princeton, NJ (US)
Assigned to INBIOSE N.V., Zwijnaarde (BE)
Appl. No. 17/049,814
Filed by INBIOSE N.V., Zwijnaarde (BE)
PCT Filed Apr. 23, 2018, PCT No. PCT/US2018/028829
§ 371(c)(1), (2) Date Oct. 22, 2020,
PCT Pub. No. WO2019/209241, PCT Pub. Date Oct. 31, 2019.
Prior Publication US 2021/0238574 A1, Aug. 5, 2021
Int. Cl. C12N 9/88 (2006.01); C12N 1/20 (2006.01); C12N 9/04 (2006.01); C12N 9/10 (2006.01); C12P 19/18 (2006.01)
CPC C12N 9/88 (2013.01) [C12N 1/20 (2013.01); C12N 9/0006 (2013.01); C12N 9/1051 (2013.01); C12P 19/18 (2013.01); C12Y 101/01 (2013.01); C12Y 204/01069 (2013.01); C12Y 402/01047 (2013.01)] 17 Claims
 
1. 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.