Production of xylitol from a mixture of hemicellulosic sugars
Materials and methods are described to produce xylitol from a mixture of hemicellulosic sugars by several routes. Examples include either as a direct co-product of a biorefinery or ethanol facility, or as a stand-alone product produced from an agricultural or forestry biomass feedstock including using, e.g. ethanol waste streams.
1. A method to produce xylitol from a mixture of hemicellulosic sugars having a xylose:arabinose ratio of about 3:1 or better, the method comprising treating the mixture of hemicellulosic sugars with enzymes produced by genetically engineered microorganisms, wherein the genetically engineered microorganisms comprise microorganisms having a recombinant xylose reductase (XR) that is more active on xylose than arabinose, wherein the genetically engineered microorganisms comprise an inactivated ptsG gene and an inactivated xylB gene, wherein arabinose and xylose are consumed, and wherein the enzymes produce xylitol at an increased purity with little or no arabinitol present in a final mixture.
2. The method to produce xylitol of claim 1 , wherein the recombinant xylose reductase is from Chaetomium globosum or Neurospora crassa.
3. The method to produce xylitol of claim 1 , comprising reducing D-xylose and metabolizing arabinose.
4. The method of claim 1 , wherein the recombinant xylose reductase has the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, or SEQ ID NO:20.
5. The method of claim 1 , wherein the mixture of hemicellulosic sugars is corn fiber hydrolysate, corn stover hydrolysate, bagasse hydrolysate, stillage hydrolysate, wheat straw hydrolysate, hardwood hydrolysate, softwood hydrolysate or other biomass sources.
6. The method of claim 1 , wherein arabinitol is less than 10% of the final mixture.
7. The method of claim 1 , wherein arabinitol is less than 5% of the final mixture.
8. The method of claim 1 , wherein the genetically engineered microorganisms are selected from the group consisting of ZUC220, ZUC136, and combinations thereof.
9. The method of claim 1 , wherein xylitol is recovered from the final mixture.
10. The method of claim 1 , wherein the mixture of hemicellulosic sugars is also treated with calcium hydroxide.
11. A method to produce xylitol from a mixture of hemicellulosic sugars having a xylose:arabinose ratio of about 3:1 or better, the method comprising treating the mixture of hemicellulosic sugars with enzymes produced by one or more genetically engineered microorganisms selected from the group consisting of ZUC220, ZUC136, and combinations thereof, wherein the genetically engineered microorganisms comprise microorganisms having a recombinant xylose reductase (XR) that is more active on xylose than arabinose, wherein arabinose and xylose are consumed, wherein the enzymes produce xylitol at an increased purity with little or no arabinitol present in a final mixture.
12. The method of claim 1 , wherein the mixture of hemicellulosic sugars comprise non-synthetic hemicellulose.
13. The method of claim 12 , wherein the non-synthetic hemicellulose is derived from corn fiber, woody fiber, hardwood hydrolysate, corn stover, corn cob, bagasse, stillage, wheat straw, hardwood, or softwood.
14. The method of claim 1 , wherein the xylitol is produced by fermentation and wherein little or no arabinitol is present in a final fermentation broth.
15. The method of claim 1 , wherein the recombinant xylose reductase has the amino acid sequence set forth in SEQ ID NO:4.
16. The method of claim 1 , wherein the genetically engineered microorganisms further comprise an inactivated lyxK gene.
17. The method of claim 1 , wherein the genetically engineered microorganisms are selected on M9+glucose liquid medium several times for improved growth.
18. The method to produce xylitol of claim 1 , wherein the recombinant xylose reductase is from Chaetomium globosum.