# US 11,078,501 B2 · Methods of fermenting mixtures that include di- and tri-saccharides formed at low temperature using a maltophilic yeast

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- Number: US 11,078,501 B2
- Document ID: US11078501B2
- Status: Granted
- Kind: B2
- Application number: 16/235,119
- Filed: 2018-12-28
- Published: 2019-07-04
- Granted: 2021-08-03
- Earliest claimed: 2017-12-29
- Assignees: Xylogenics, Inc.
- Applicants: Xylogenics, Inc.
- Inventors: Joshua W. Heyen; Mark G. Goebl; Kathryn A. Houin; Matthew S. Kelker; Nadaraj Palaniappan
- Web page: https://patentgrep.com/patent/US11078501B2

## CPC

- C12P 7/06: Ethanol, i.e. non-beverage
- 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)}
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

Disclosed herein are materials and methods for the production of high concentrations of ethanol from plant starch material broken down into disaccharide and trisaccharide sugars such as maltose, isomaltose, and maltotriose from uncooked starch. Herein is also described use of a yeast strain capable of fermenting high maltose syrups into ethanol without the need to convert disaccharides or trisaccharides into glucose using exogenous glucoamylases.

## Claims

### Claim 1

A method for producing ethanol from a plant matter, comprising the steps of:
liquefying of grain solids by treating a combination of the grain solids and water with an alpha-amylase at a concentration between 0.5 to 4 wt. % of the grain solids at a temperature of 70° C. or lower to form a grain slurry;
adding a maltogenic alpha-amylase; a maltophilic strain of Saccharomyces; and beta amylase to the grain slurry to form a mixture; and
fermenting the mixture at a temperature of about 38° C. or lower,
wherein the maltophilic strain of Saccharomyces is a GRR1 knock-out of Ethanol Red.

### Claim 2 (depends on 1)

The method according to claim 1, further including the step of adding a pullulanase to the mixture.

### Claim 3 (depends on 1)

The method according to claim 1, further including the step of adding a second amount of an alpha amylase to the mixture during the fermentation step.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the method is carried out in the absence of exogenous glucoamylases.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the grain slurry is formed by treating a combination of the grain solids and water with an alpha-amylase at a concentration between 0.5 to 4 wt. % of the grain solids at a temperature in the range of about 50° C. to 70° C.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the grain slurry is adjusted to a temperature of less than about 30° C. before the maltophilic strain of Saccharomyces is contacted with the grain slurry.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the grain slurry of the plant matter comprises a portion of grain solids having grain solids content of between about 28 weight percent (wt. %) to about 36 (wt. %) grain solids.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the content of alpha amylases in the method is between about 2.0 wt. % grain solids to about 4 wt. % grain solids.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the content of beta amylases in the method is between about 0.5 wt. % grain solids to about 4 wt. % grain solids.

### Claim 10 (depends on 9)

The method according to claim 9, wherein the content of beta amylases in the method is between about 2.0 wt. % grain solids to about 4 wt. % grain solids.

### Claim 11 (depends on 2)

The method according to claim 2, wherein the content of pullulanase in the method is between about 0.5 wt. % grain solids to about 4 wt. %.

### Claim 12 (depends on 11)

The method according to claim 11, wherein the content of pullulanase in the method is between about 2.0 wt. % grain solids to about 4 wt. % grain solids.

### Claim 13 (depends on 1)

The method of claim 1, wherein the grain is corn.