Methods for using phytase in ethanol production
The present disclosure relates to methods for using one or more polypeptides with phytase activity in grain processing, ethanol, and biofuel production.
1. A method for improving efficiency of ethanol production by fermentation in an ethanol production facility, said method comprising:
adding a phytase variant polypeptide comprising an amino acid sequence as set forth in amino acid residues 23-434 of SEQ ID NO: 2 and having amino acid substitutions M298K and A299T
to an ethanol processing fluid in the ethanol production facility, wherein the ethanol processing fluid comprises phytic acid, wherein the addition of the phytase variant polypeptide improves ethanol production efficiency comprising one or more of:
increased yeast cell count, yeast budding or yeast viability;
increased ethanol yield, decreased glycerol levels, or decreased total sugar levels;
decreased fouling rate;
increased phosphorous levels; and
increased operation time of the ethanol production facility when compared to the ethanol production process wherein a wild-type phytase consisting of the amino acid sequence as set forth in amino acid residues 23-434 of SEQ ID NO: 2 is added to the ethanol processing fluid.
2. The method of claim 1 , wherein addition of the phytase variant polypeptide improves ethanol production efficiency compared to a production process where a commercially available phytase is added, and wherein the commercially available phytase is selected from Novozyme 50161™ and U.S. Water PhytOUT™.
3. The method of claim 1 , wherein the ethanol production facility includes a hammer mill for grinding feedstock, a slurry tank, a jet cooker, a liquefaction tank, a mash cooker, a yeast mix tank, a yeast propagator, a fermentation tank, a beer, a distillation system, centrifuge, an evaporator, a condensate, a syrup, a wet grain, a drum dryer, or any combination thereof.
4. The method of claim 3 , wherein the ethanol production facility comprises a mash cooker and the phytase variant polypeptide is added to the ethanol processing fluid in the mash cooker.
5. The method of claim 3 , wherein the ethanol production facility comprises a yeast mix tank and a Yeast propagator and the phytase variant polypeptide is added to the ethanol processing fluid in a yeast mix tank.
6. The method of claim 5 , wherein the ethanol processing fluid proceeds from the yeast mix tank to ale yeast propagator.
7. The method of claim 3 , wherein the wherein the ethanol production facility comprises a fermentation tank and a distillation system, wherein said fermentation tank comprises Yeast cells which convert sugar, starch, or cellulose to ethanol, and the ethanol produced in the fermentation tank is separated by the distillation system.
8. The method of claim 1 , wherein the phytase variant polypeptide is added to the ethanol processing fluid prior to fermentation.
9. The method of claim 1 , wherein the phytase variant polypeptide hydrolyzes phytic acid to inositol and free phosphate with release of minerals from the phytic acid.
10. The method of claim 9 , wherein the phytic acid is phytate in the form of calcium salts, magnesium salts, metal ions, proteins, unhydrolyzed phytate sludge, or myo-inositol-hexaphosphate.
11. The method of claim 1 , wherein the phytase variant polypeptide is used in combination with one or more other enzymes.
12. The method of claim 11 , wherein the one or more other enzymes comprise an amylase, a glucoamylase, a glucanase, a cellulase, an endoglucanase, a mannase, a xylanase, a xanthanase, a glycosidases, a cellobiohydrolase, a beta-glucosidase, a pullanase, a glucoisomerase, an alpha-glucosidase, or a combination thereof.
13. The method of claim 1 , wherein the phytase variant polypeptide retains phytase activity under conditions from pH 2.5 to pH 12.0.
14. The method of claim 1 , wherein the ethanol is produced from a feedstock is selected from the group consisting of corn, wheat, barley, potatoes, switchgrass, Miscanthus, poplar wood, rice straw, corn stover, wheat straw, sugarcane bagasse, rice hulls, corn fiber, sugar beet pulp, citrus pulp, citrus peels, hardwood, softwood thinnings, hardwood and softwood residues from timber operations, paper traction of municipal solid waste, municipal wood waste, municipal green waste, saw mill waste, pulp mill waste, construction waste, demolition waste, wood shavings, sawdust, waste paper, materials containing sugar, starch, and cellulose.
15. The method of claim 1 , wherein the ethanol production facility is in an ethanol production plant; a spirit or a drinkable alcohol production plant; or a fuel ethanol plant.