# US 10,246,718 B2 · Processes for producing lipids

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- Number: US 10,246,718 B2
- Document ID: US10246718B2
- Status: Granted
- Kind: B2
- Application number: 15/293,032
- Filed: 2016-10-13
- Published: 2017-02-02
- Granted: 2019-04-02
- Earliest claimed: 2011-12-27
- Assignees: Commonwealth Scientific and Industrial Research Organisation
- Applicants: Commonwealth Scientific and Industrial Research Organisation
- Inventors: Thomas Vanhercke; James Robertson Petrie; Anna El Tahchy; Surinder Pal Singh; Qing Liu
- Web page: https://patentgrep.com/patent/US10246718B2

## CPC

- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- A23K 10/30: from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms (obtained by microbiological or biochemical processes, e.g. using yeasts or enzymes, A23K10/10)
- A23K 20/153: Nucleic acids; Hydrolysis products or derivatives thereof
- C07K 14/415: from plants
- C10G 2/00: Production of liquid hydrocarbon mixtures of undefined composition from oxides of carbon
- C10L 1/026: {for compression ignition}
- C11B 1/04: of vegetable raw material {(C11B1/025 takes precedence)}
- C11B 1/10: by extracting
- C11B 3/001: {by a combination of two or more of the means hereafter}
- C11B 3/006: {by extraction}
- C11B 3/10: by adsorption
- C11B 3/14: with the use of indifferent gases or vapours, e.g. steam
- C11B 7/0008: {by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents (addition of a minor amount of solvent intended to lower the melting point C11B7/0083, C11B7/0091)}
- C11B 7/0075: {by differences of melting or solidifying points}
- C11C 3/003: {by esterification of fatty acids with alcohols (C11C3/02 takes precedence)}
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- C10L 5/44: on vegetable substances
- C10L 2200/0476: Biodiesel, i.e. defined lower alkyl esters of fatty acids first generation biodiesel
- C10L 2290/24: Mixing, stirring of fuel components
- C10L 2290/544: Extraction for separating fractions, components or impurities during preparation or upgrading of a fuel
- Y02E 50/30: Fuel from waste, e.g. synthetic alcohol or diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A23K: FODDER
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C10G: CRACKING HYDROCARBON OILS; PRODUCTION OF LIQUID HYDROCARBON MIXTURES, e.g. BY DESTRUCTIVE HYDROGENATION, OLIGOMERISATION, POLYMERISATION (cracking to hydrogen or synthesis gas C01B; cracking or pyrolysis of hydrocarbon gases to individual hydrocarbons or mixtures thereof of definite or specified constitution C07C; cracking to cokes C10B); RECOVERY OF HYDROCARBON OILS FROM OIL-SHALE, OIL-SAND, OR GASES; REFINING MIXTURES MAINLY CONSISTING OF HYDROCARBONS; REFORMING OF NAPHTHA; MINERAL WAXES
- C10L: FUELS NOT OTHERWISE PROVIDED FOR; NATURAL GAS; SYNTHETIC NATURAL GAS OBTAINED BY PROCESSES NOT COVERED BY SUBCLASSES C10G OR C10K; LIQUIFIED PETROLEUM GAS; USE OF ADDITIVES TO FUELS OR FIRES; FIRE-LIGHTERS
- C11B: PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES (drying-oils C09F)
- C11C: FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- 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

The present invention relates to processes for extracting lipid from vegetative plant parts such as leaves, stems, roots and tubers, and for producing industrial products such as hydrocarbon products from the lipids. Preferred industrial products include alkyl esters which may be blended with petroleum based fuels.

## Claims

### Claim 1

A process for feeding an animal, comprising a step of providing to the animal a vegetative plant part from a transformed plant, the vegetative plant part comprising one or more exogenous polynucleotides which encode at least Wrinkled 1 (WRI1) and a fatty acid acyltransferase and having a total non-polar lipid content of at least 10% (w/w dry weight).

### Claim 2 (depends on 1)

The process of claim 1, wherein the vegetative plant part is a plant leaf.

### Claim 3 (depends on 2)

The process of claim 2, wherein the plant leaf comprises a total TAG content of about 11% (w/w dry weight).

### Claim 4 (depends on 2)

The process of claim 2, wherein the total fatty acid content in the non-polar lipid of the plant leaf comprises at least 2% more oleic acid than the non-polar lipid in a corresponding wild-type plant leaf.

### Claim 5 (depends on 2)

The process of claim 2, wherein the total fatty acid content in the non-polar lipid of the plant leaf comprises at least 2% less palmitic acid than the non-polar lipid in a corresponding wild-type plant leaf.

### Claim 6 (depends on 2)

The process of claim 2, wherein the non-polar lipid of the plant leaf comprises a modified level of total sterols, non-esterified sterols, steroyl esters or steroyl glycosides relative to the non-polar lipid in a corresponding wild-type plant leaf.

### Claim 7 (depends on 1)

The process of claim 1, wherein the vegetative plant part has one or more or all of the following features:
i) oleic acid comprises at least 19% of the total fatty acid content in the non-polar lipid in the vegetative plant part,
ii) palmitic acid comprises at least 20% of the total fatty acid content in the non-polar lipid in the vegetative plant part,
iii) linoleic acid comprises at least 15% of the total fatty acid content in the non-polar lipid in the vegetative plant part, and
iv) α-linolenic acid comprises less than 15% of the total fatty acid content in the non-polar lipid in the vegetative plant part.

### Claim 8 (depends on 7)

The process of claim 7, wherein oleic acid comprises at least 19% of the total fatty acid content in the non-polar lipid in the vegetative plant part.

### Claim 9 (depends on 8)

The process of claim 8, wherein the vegetative plant part is a plant leaf.

### Claim 10 (depends on 1)

The process of claim 1, wherein the vegetative plant part has a total non-polar content of about 11% (w/w dry weight).

### Claim 11 (depends on 10)

The process of claim 10, wherein the vegetative plant part is a plant leaf.

### Claim 12 (depends on 1)

The process of claim 1, wherein the vegetative plant part has a total non-polar lipid content of about 15% (w/w dry weight).

### Claim 13 (depends on 1)

The process of claim 1 which comprises a step of harvesting the vegetative plant part from a plant grown in the field with a mechanical harvester.

### Claim 14 (depends on 13)

The process of claim 13, wherein the vegetative plant part is harvested from the plant at a time between about the time of flowering of the plant to about the time senescence of the plant has started.

### Claim 15 (depends on 1)

The process of claim 1, wherein the fatty acid acyltransferase is a phospholipid:diacylglycerol acyltransferase.

### Claim 16 (depends on 1)

The process of claim 1, wherein the fatty acid acyltransferase is a monoacylglycerol acyltransferase (MGAT), a diacylglycerol acyltransferase (DGAT), or a glycerol-3-phosphate acyltransferase (GPAT).