Manufacture of five-carbon sugars and sugar alcohols
View Patent ↗The invention relates to the methods of manufacturing five-carbon sugars and sugar alcohols as well as other compounds derived from pentose-phosphate pathway from readily available substrates such a hexoses using metabolically engineered microbial hosts.
1. A method for the production of xylitol, said method comprising:
(A) cultivating a genetically modified xylulose-5-phosphate producing, bacterial, yeast or fungal host, the genetic modification of which increases the expression of xylitol phosphate dehydrogenase in said host during said cultivating as compared to said activity in said host prior to being genetically modified, on a carbon source other than D-xylose, D-xylulose, mixtures of D-xylose and D-xylulose, and polymers and oligomers containing D-xylose or D-xylulose as major components, wherein said modification comprises introducing one or more genes encoding said xylitol phosphate dehydrogenase to said host wherein said xylitol phosphate dehydrogenase is L. rhamnosus xylitol 1-phosphate dehydrogenase or B. halodurans xylitol 1-phosphate dehydrogenase. or C. difficile xylitol 1-phosphate dehydrogenase;
(B) producing xylitol during said cultivating of part (A) by using said host to convert one or more pentose phosphate metabolic pathway intermediates in said host into said xylitol; and
(C) recovering said xylitol that is produced in part (B);
wherein the amount or rate of said xylitol production in said genetically modified microbial host is enhanced as compared to said amount or rate of xylitol production in said host prior to being said genetically modified.
2. The method of claim 1 , wherein said xylitol phosphate dehydrogenase is L. rhamnosus xylitol 1-phosphate dehydrogenase.
3. The method of claim 2 , wherein said. L. rhamnosus xylitol phosphate dehydrogenase comprises the amino acid sequence of SEQ ID NO:49.
4. The method of claim 3 , wherein said L. rhamnosus xylitol phosphate dehydrogenase is encoded by a gene that comprises the nucleic acid sequence of SEQ ID NO:48.
5. The method of claim 1 , wherein said xylitol phosphate dehydrogenase is B. halodurans xylitol 1-phosphate dehydrogenase.
6. The method of claim 5 , wherein said B. halodurans xylitol phosphate dehydrogenase comprises the amino acid sequence of SEQ ID NO:50.
7. The method of claim 1 , wherein said xylitol phosphate dehydrogenase is a C. difficile xylitol 1-phosphate dehydrogenase.
8. The method of claim 7 , wherein said C. difficile xylitol phosphate dehydrogenase comprises the amino acid sequence of a sequence selected from the group consisting of SEQ ID NOs:51, 52 and 53.