# US 12,110,531 B2 · Method for increasing oil yield during ethanol production

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- Number: US 12,110,531 B2
- Document ID: US12110531B2
- Status: Granted
- Kind: B2
- Application number: 16/621,957
- Filed: 2018-06-14
- Published: 2023-05-25
- Granted: 2024-10-08
- Earliest claimed: 2017-06-16
- Assignees: Danstar Ferment AG
- Applicants: Danstar Ferment AG
- Inventors: Brian Hoskins; Anthony V. Newton; Yun Han; Joseph P. Borst
- Web page: https://patentgrep.com/patent/US12110531B2

## CPC

- C12P 7/64: Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12N 9/2414: {Alpha-amylase (3.2.1.1.)}
- C12P 7/06: Ethanol, i.e. non-beverage
- C12Y 302/01001: Alpha-amylase (3.2.1.1)
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

A method for increasing the oil yield in an ethanol production process comprising: adding a liquid enzyme formulation having at least one enzyme, a buffering agent, a stabilizer, and a preservative wherein the pH of the enzyme formulation is about pH 6.0-8.0 to a beer, a distillation, a whole stillage, a centrifugation, a thin stillage, an evaporator, a syrup, or an oil recovery unit.

## Claims

### Claim 1

A method for increasing the oil yield in an ethanol production process comprising: adding a liquid enzyme formulation having an alpha-amylase titrated from a pH of 8.0 to 10.5, a buffering agent, a stabilizer, and a preservative wherein the pH of the enzyme formulation is about pH 6.0-8.0 to a beer, a distillation, a whole stillage, a centrifugation, a thin stillage, an evaporator, a syrup, or an oil recovery unit, and wherein the alpha-amylase comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of the amino acid sequences 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, and SEQ ID NO:14.

### Claim 2 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the pH of the liquid enzyme formulation is about pH 6.3-6.7.

### Claim 3 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the stabilizer comprises sucrose, sorbitol, mannitol, glycerol, trehalose, sodium chloride, sodium sulfate, or any combination thereof.

### Claim 4 (depends on 1)

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.

### Claim 5 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the alpha-amylase retains at least 90% of its activity at a temperature of 4-40° C.

### Claim 6 (depends on 1)

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.

### Claim 7 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the alpha-amylase retains at least 90% of its activity for 1 year.

### Claim 8 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the alpha-amylase has a shelf life of at least 1 year.

### Claim 9 (depends on 8)

The liquid enzyme formulation of claim 8, wherein the alpha-amylase has a shelf life of at least 1 year at 25° C.

### Claim 10 (depends on 1)

The liquid enzyme formulation of claim 1, wherein the preservative comprises: potassium sorbate, sodium sorbate, sorbic acid, sodium benzoate, benzoic acid, methyl paraben, calcium propionate, sodium propionate, ammonium propionate, propionic acid, or any combination thereof.

### Claim 11 (depends on 1)

The liquid enzyme formulation of claim 1, further comprising at least two preservatives.

### Claim 12 (depends on 1)

The liquid enzyme formulation of claim 1, further comprising a second enzyme.

### Claim 13 (depends on 12)

The liquid enzyme formulation of claim 12, wherein the second enzyme is selected from the group consisting of a second 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, a lipase, a phospholipase, and any combination thereof.

### Claim 14 (depends on 1)

The method of claim 1, wherein a liquid enzyme formulation having at least one enzyme is added to a first stillage composition to form a second stillage composition, wherein the one or more alpha amylase enzymes are added to the first stillage composition in an amount from 0.001 to 0.01 grams/100 grams of solids of the first stillage composition; and obtaining oil from the second stillage composition.

### Claim 15 (depends on 14)

The method of claim 14, wherein the first stillage composition comprises whole stillage, thin stillage, wet cake and/or syrup.