Methods for using a thermostable phytase in ethanol production

1. A method for eliminating or reducing phytic acid and salts of phytic acid in an alcohol production process comprising:

(a) providing a variant polypeptide comprising an amino acid sequence that is at least 95% identical to the amino acid sequence as set forth in SEQ ID NO: 2 and one single amino acid substitution to the amino acid sequence of SEQ ID NO: 2, wherein said one single amino acid substitution is selected from the group consisting of: A47F, T136F, N159V, N159E, T163R, D164R, G179R, V233W, Q275V, R289A, and T349Y; wherein said variant polypeptide has phytase activity and;

(b) providing a composition comprising a phytic acid or a salt of phytic acid; and

(c) contacting said variant polypeptide of (a) with said composition of (b), wherein said variant polypeptide hydrolyzes the phytic acid or salt of phytic acid.

2. The method of claim 1 , wherein said variant polypeptide having phytase activity is added anywhere in the alcohol production process wherein said alcohol production process comprises a feedstock, a hammer mill, a slurry tank, a jet cooker, a liquefaction, a mash cooker, a fermentation, a beer, a distillation system, a whole stillage, a centrifuge, a thin stillage, an evaporator, a condensate, a syrup, a wet grain, a drum dryer, a dried distiller's grains with solubles, a condensed distiller's solubles, a dried distiller's grain, a wet distiller's grains with solubles, or any combination thereof.

3. The method of claim 1 , wherein said alcohol production process is in an ethanol production plant; a spirit or a drinkable alcohol production plant; or a fuel ethanol plant.

4. The method of claim 2 , wherein said 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, wood shavings, sawdust, paper fraction 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/or cellulose.

5. The method of claim 2 , wherein the fermentation converts sugar, starch, or cellulose to alcohol with a yeast, and the alcohol is separated with distillation.

6. The method of claim 1 , wherein said variant polypeptide having phytase activity hydrolyzes phytate to inositol and free phosphate with release of minerals from the phytic acid.

7. The method of claim 1 , wherein the phytic acid is phytate in the form of calcium salts, magnesium salts, metal ions, proteins, unhydrolyzed phytate sludge, or myo-inositol-hexaphosphate.

8. The method of claim 1 , wherein said variant polypeptide having phytase activity can be used in combination with one or more other enzymes.

9. The method of claim 8 , wherein said one or more other enzymes comprises 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, a alpha-glucosidase.

10. The method of claim 1 , wherein said variant polypeptide having phytase activity retains phytase activity under conditions from pH 2.5 to pH 12.0.

11. The method of claim 1 , wherein said variant polypeptide having phytase activity improves production of ethanol from starch.