Cell-based production of nonulosonates
AssigneeNational Research Council of Canada
InventorsDennis M. Whitfield, Susan M. Logan, Ian C. Schoenhofen, Christopher N. Boddy, Benjamin R. Lundgren
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Abstract
The present invention relates to the cell-based production of bacterial nonulosonates and their biosynthetic precursors. Specifically, the present invention provides recombinant cells for the production of pseudaminic acid, legionaminic acid, UDP-2,4-diacetamido-2,4,6-trideoxy-β-L-altropyranose, and UDP-2,4-diacetamido-2,4,6-trideoxy-α-D-glucopyranose. Methods for producing the sugars are also provided.
Claims19
A recombinant cell for the production of legionaminic acid, the recombinant cell comprising an inactivated GlcNAc-6-P deacetylase gene;
- a gene encoding a UDP-GlcNAc 4,6-dehydratase that is capable of converting UDP-GlcNAc into UDP-2-acetamido-2,6-dideoxy-α-D-xylo-hexos-4-ulose;
- a gene encoding an aminotransferase that is capable of converting UDP-2-acetamido-2,6-dideoxy-α-D-xylo-hexos-4-ulose into UDP-4-amino-4,6-dideoxy-α-D-GlcNAc or UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-Glc;
- a gene encoding an N-acetyltransferase that is capable of converting UDP-4-amino-4,6-dideoxy-α-D-GlcNAc or UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-Glc into UDP-2,4-diacetamido-2,4,6-trideoxy-α-D-Glc (UDP-BacdiNAc);
- a gene encoding a Leg synthase that is capable of converting 2,4-diacetamido-2,4,6-trideoxy-D-Man (6-deoxy-MandiNAc) into 5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto-nonulosonic acid (Leg);
- a gene encoding a UDP-sugar hydrolase and 2-epimerase that is capable of converting UDP-2,4-diacetamido-2,4,6-trideoxy-α-D-Glc (UDP-BacdiNAc) into 2,4-diacetamido-2,4,6-trideoxy-D-Man (6-deoxy-MandiNAc);
- a gene encoding a GlcNAc-6-P mutase;
- and a gene encoding a GlcNAc-1-P uridyltransferase.
A recombinant cell for the production of UDP-2,4-diacetamido-2,4,6-trideoxy-α-D-glucopyranose (UDP-BacdiNAc), comprising an inactivated GlcNAc-6-P deacetylase gene;
- a gene encoding a UDP-GlcNAc 4,6-dehydratase that is capable of converting UDP-GlcNAc into UDP-2-acetamido-2,6-dideoxy-α-D-xylo-hexos-4-ulose;
- a gene encoding an aminotransferase that is capable of converting UDP-2-acetamido-2,6-dideoxy-α-D-xylo-hexos-4-ulose into UDP-4-amino-4,6-dideoxy-α-D-GlcNAc or UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-Glc;
- a gene encoding an N-acetyltransferase that is capable of converting UDP-4-amino-4,6-dideoxy-α-D-GlcNAc or UDP-2-acetamido-4-amino-2,4,6-trideoxy-α-D-Glc into UDP-2,4-diacetamido-2,4,6-trideoxy-α-D-Glc (UDP-BacdiNAc);
- a gene encoding a GlcNAc-6-P mutase;
- and a gene encoding a GlcNAc-1-P uridyltransferase.
Dependencies are inferred from the claim text.