# US 9,434,962 B2 · Integrated process for producing biofuels

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- Number: US 9,434,962 B2
- Document ID: US9434962B2
- Status: Granted
- Kind: B2
- Application number: 13/333,195
- Filed: 2011-12-21
- Published: 2012-06-28
- Granted: 2016-09-06
- Earliest claimed: 2010-12-22
- Assignees: Neste Oyj
- Applicants: Perttu Koskinen; Reijo Tanner
- Inventors: Perttu Koskinen; Reijo Tanner
- Web page: https://patentgrep.com/patent/US9434962B2

## CPC

- C12P 7/16: Butanols
- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12N 9/248: {Xylanases}
- C12P 7/10: substrate containing cellulosic material
- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- Y02E 50/10: Biofuels, e.g. bio-diesel
- Y02P 20/10: Process efficiency
- Y02P 20/52: using catalysts, e.g. selective catalysts
- Y02P 20/582: Recycling of unreacted starting or intermediate materials
- 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)
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

Disclosed is an integrated process comprising first and second biotechnical processes. The first process produces biofuel and/or starting material for biofuel and uses a microorganism capable of producing enzymes. The second process produces biofuel and/or starting material for biofuel. In the integrated process, the microorganisms are cultivated and biofuel and/or starting material for biofuel and enzymes are produced. The microorganism culture, supernatant or a protein enriched fraction or a dilution of the supernatant comprising catalytically active enzyme(s) are introduced into the first and/or into the second process, or feedstock for the process(es) is treated. The invention relates also to use of the produced enzymes in biofuel production process or in other applications as an enzyme preparation or a source of enzymes. The invention relates also to use of the produced lipids and alcohols as biofuel, a component of biofuel or a starting material for biofuel production.

## Claims

### Claim 1

An integrated process for single cell oil production and alcohol production, the process comprising steps:
(a) cultivating a fungus belonging to the genus Aspergillus in a culture media in a single cell oil production process, wherein the culture media comprises hemicellulose and the fungus produces single cell oil and xylanase; and
(b) cultivating a microorganism in a culture media in an alcohol production process, wherein the microorganism produces alcohol and the culture media comprises:
(i) lignocellulosic material or fractions thereof, and
the xylanase produced in step (a) or a fraction of the microorganism culture of step (a) that comprises the xylanase;
or
(ii) treated lignocellulosic material or fractions thereof;
wherein the treated lignocellulosic material or fractions thereof have been treated with the xylanase produced in step (a), or
wherein the treated lignocellulosic material or fractions thereof have been treated with a fraction of the microorganism culture of step (a) that comprises the xylanase.

### Claim 2 (depends on 1)

The process according to claim 1, wherein the alcohol is ethanol, butanol, isopropanol, or mixtures thereof.

### Claim 3 (depends on 1)

The process according to claim 1, further comprising separating a supernatant and the fungus cells from the culture of the single cell oil production process of step (a), and introducing the separated supernatant or the separated fungus cells to the culture media of the alcohol production process of step (b).

### Claim 4 (depends on 1)

The process according to claim 1, further comprising separating a supernatant and the fungus cells from the culture of the single cell oil production process of step (a), treating lignocellulosic material or fractions thereof with the separated supernatant or the separated fungus cells, and introducing the treated lignocellulosic material or fractions thereof to the culture media of the alcohol production process of step (b).

### Claim 5 (depends on 1)

The process according to claim 1, further comprising recovering the xylanase produced in the single cell oil production process of step (a) and introducing the recovered xylanase to the culture media of the alcohol production process of step (b).

### Claim 6 (depends on 1)

The process according to claim 1, further comprising recovering the xylanase produced in the single cell oil production process of step (a), treating lignocellulosic material or fractions thereof with the recovered xylanase, and introducing the treated lignocellulosic material or fractions thereof to the culture media in the alcohol production process of step (b).

### Claim 7 (depends on 1)

The process according to claim 1, wherein supernatant from the fungus culture of step (a) comprises the xylanase, and the culture media of step (b) comprises the supernatant, a dilution thereof, or a protein-enriched fraction thereof.

### Claim 8 (depends on 1)

The process according to claim 1, further comprising recovering the single cell oil of the single cell oil production process of step (a) using a recovery method that preserves the catalytic activity of the xylanase produced in the single cell oil production process of step (a).

### Claim 9 (depends on 1)

The process according to claim 1, wherein the culture media in the alcohol production process of step (b) further comprises starch.

### Claim 10 (depends on 1)

The process according to claim 1, wherein the fungus in the single cell oil production process of step (a) further produces enzymes selected from the group consisting of hemicellulases, cellulases, mannanases, arabinases, galactosidases, glucosidases, mannosidases, xylosidases, arabinofuranosidase, esterases, endo-cellulases, exo-cellulases, cellobiases, beta-glucosidases, oxidative cellulases, cellulose phosphorylases and mixtures thereof.

### Claim 11 (depends on 1)

The process according to claim 1, further comprising recovering the xylanase of the single cell oil production process of step (a), and recycling the recovered xylanase to culture media in a single cell oil production process.

### Claim 12 (depends on 1)

The process according to claim 1, further comprising recovering lipids from the single cell oil production process of step (a) and reacting the lipids with alcohol in a transesterification reaction to produce biodiesel.

### Claim 13 (depends on 1)

The process according to claim 1, further comprising recovering alcohol from the alcohol production process of step (b) and reacting the alcohol with lipids in a transesterification reaction to produce biodiesel.