Production of L-ribose and other rare sugars
The invention provides methods and compositions for the production of L-ribitol and other rare sugars using a mannitol-1-dehydrogenase.
1. A method for producing L-ribose comprising fermenting a substrate comprising ribitol with one or more recombinant bacteria or yeast that expresses a recombinant Apium graveolens mannitol-1-dehydrogenase (MDH), wherein (1) the recombinant Apium graveolens MDH has a greater reaction rate than native MDH or (2) the one or more recombinant bacteria or yeast convert approximately 25% of ribitol to L -ribose within 48 hours, whereby L-ribose is produced.
2. The method of claim 1 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) is encoded by a nucleic acid molecule comprising SEQ ID NO:1.
3. The method of claim 1 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) comprises SEQ ID NO:2.
4. The method of claim 1 , wherein the one or more recombinant bacteria or yeast express ribitol transport proteins (rbT).
5. The method of claim 1 , wherein the substrate is further contacted with a NADH oxidase.
6. The method of claim 5 , wherein the NADH oxidase is NOX from Lactobacillus sanfranciscensis.
7. A method for producing L-ribose from a substrate comprising D-ribose comprising:
(a) converting D-ribose to ribitol; and
(b) fermenting the ribitol with one or more recombinant bacteria or yeast that express a recombinant Apium graveolens mannitol-1-dehydrogenase (MDH), wherein (1) the recombinant Apium graveolens MDH has a greater reaction rate than native MDH or (2) the one or more recombinant bacteria or yeast convert approximately 25% of ribitol to L -ribose within 48 hours, whereby L-ribose is produced.
8. The method of claim 7 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) is encoded by a nucleic acid molecule comprising SEQ ID NO:1.
9. The method of claim 7 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) comprises SEQ ID NO:2.
10. The method of claim 7 , wherein the one or more recombinant bacteria or yeast express ribitol transport proteins (rbT).
11. The method of claim 7 , wherein the ribitol is further contacted with a NADH oxidase.
12. The method of claim 11 , wherein the NADH oxidase is NOX from Lactobacillus sanfranciscensis.
13. The method of claim 7 , wherein the D-ribose is converted to ribitol by chemical reduction.
14. The method of claim 7 , wherein D-ribose is converted to ribitol by fermentation.
15. A method for producing a sugar from a substrate comprising fermenting the substrate with one or more recombinant bacteria or yeast that express a recombinant Apium graveolens mannitol-1-dehydrogenase (MDH), wherein the recombinant Apium graveolens MDH has a greater reaction rate than native MDH, whereby the sugar is produced, wherein the sugar is D-arabanose and the substrate is D-arabitol; wherein the sugar is L-erythrose and the substrate is erythritol; wherein the sugar is L-gulose and the substrate is L-sorbitol; wherein the sugar is L-galactose and the substrate is i-galactitol; wherein the sugar is D-threose and the substrate is D-threitol; wherein the sugar is L -fucose and the substrate is L -fucitol; or wherein the sugar is D-lyxose and the substrate is D-arabitol.
16. The method of claim 15 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) is encoded by a nucleic acid molecule comprising SEQ ID NO:1.
17. The method of claim 15 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) comprises SEQ ID NO:2.
18. The method of claim 15 , wherein the one or more recombinant bacteria or yeast express ribitol transport proteins (rbT).
19. The method of claim 15 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) is encoded by a nucleic acid molecule having optimized codon usage for the one or more recombinant bacteria or yeast used for expression of the Apium graveolens mannitol-1-dehydrogenase (MDH).
20. The method of claim 15 , wherein the substrate is further contacted with a NADH oxidase.
21. The method of claim 20 , wherein NADH oxidase is NOX from Lactobacillus sanfranciscensis.
22. A method for producing D-mannose comprising fermenting a substrate comprising D-mannitol with one or more recombinant bacteria or yeast that express a recombinant Apium graveolens mannitol-1-dehydrogenase (MDH), wherein the recombinant Apium graveolens MDH has a greater reaction rate than native MDH, and contacting the substrate with a NADH oxidase, whereby D-mannose is produced.
23. The method of claim 22 , wherein the recombinant Apium graveolens mannitol-1-dehydrogenase (MDH) comprises SEQ ID NO:2 and the NADH oxidase is NOX from Lactobacillus sanfranciscensis.