Liquid formulation of alpha-amylase
The present disclosure relates to liquid enzyme formulations containing one or more alpha-amylases for use in starch processing, wherein the pH of the enzyme formulation is about pH 6.0-8.0, and methods of use thereof. The present disclosure further relates to methods of making a liquid enzyme formulation containing one or more alpha-amylase having improved stability, comprising titrating the pH of the liquid enzyme formulation to a range of pH 6.0-8.0.
1. A liquid alpha-amylase enzyme formulation comprising:
(a) an alpha-amylase having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 or SEQ ID NO: 14;
(b) a buffering agent;
(c) a stabilizer present at 30% to 50% w/v; and
(d) a preservative, wherein the preservative is potassium sorbate, sodium sorbate, sorbic acid, sodium benzoate, benzoic acid, methyl paraben, calcium propionate, sodium propionate, ammonium propionate, propionic acid, or any combination thereof,
wherein the pH of the enzyme formulation is 6.3-6.7, and
wherein the alpha-amylase retains at least 90% of its activity at a temperature of 4-40° C. for at least one year.
2. The liquid enzyme formulation of claim 1 , wherein the stabilizer comprises sucrose, sorbitol, mannitol, glycerol, trehalose, sodium chloride, sodium sulfate, or any combination thereof.
3. The liquid enzyme formulation of claim 1 , wherein the buffering agent comprises: sodium citrate, potassium citrate, citric acid, sodium acetate, acetic acid, sodium phosphate, potassium phosphate, or any combination thereof.
4. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase retains at least 90% of its activity at a temperature of 25-30° C.
5. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase has a shelf life of at least 1 year.
6. The liquid enzyme formulation of claim 5 , wherein the alpha-amylase has a shelf life of at least 1 year at 25° C.
7. The liquid enzyme formulation of claim 1 , further comprising at least two preservatives.
8. The liquid enzyme formulation of claim 1 , further comprising a second enzyme.
9. The liquid enzyme formulation of claim 8 , wherein the second enzyme is selected from the group consisting of an alpha-amylase, a beta-amylase, a glucoamylase, a protease, a phytase, a pullulanase, a cellulase, a cellobiohydrolase, a beta-glucosidase, an endoglucanase, a mannanase, a xylanase, and any combination thereof.
10. Method of using a liquid alpha-amylase enzyme formulation, comprising:
(a) providing a liquid enzyme formulation comprising:
(i) an alpha-amylase having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 or SEQ ID NO: 14,
(ii) a buffering agent,
(iii) a stabilizer present at 30% to 50% w/v, and
(iv) a preservative,
wherein the pH value of the liquid enzyme formulation is 6.3-6.7,
wherein the preservative is potassium sorbate, sodium sorbate, sorbic acid, sodium benzoate, benzoic acid, methyl paraben, calcium propionate, sodium propionate, ammonium propionate, propionic acid, or any combination thereof;
(b) providing a starch; and
(c) contacting the liquid enzymatic formulation with the starch, thereby hydrolyzing the starch.
11. The method of claim 10 , wherein the method is used for the production of a food ingredient, a food product, a fuel, an alcohol, a cleaning product, a personal care product, an animal feed, a paper, a paper pulp, a paper product, or a starch based product.
12. The method of claim 10 , wherein the method is used in oil and gas well operations.
13. The liquid enzyme formulation of claim 1 , wherein the stabilizer is glycerol.
14. The liquid enzyme formulation of claim 1 , wherein the buffering agent is present from about 0.1% to 2% w/v.
15. The liquid enzyme formulation of claim 1 , wherein the preservative is potassium sorbate, sodium sorbate, sorbic acid, or a combination thereof.
16. The method of claim 10 , wherein the stabilizer is glycerol.
17. The method of claim 10 , wherein the buffering agent is present from about 0.1% to 2% w/v.
18. The method of claim 10 , wherein the preservative is potassium sorbate, sodium sorbate, sorbic acid, or a combination thereof.
19. The liquid enzyme formulation of claim 1 , wherein the glycerol is present at 38 to 42%.