WO2017006031A1 - Method for producing fatty acids from microorganisms - Google Patents
Method for producing fatty acids from microorganisms Download PDFInfo
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- WO2017006031A1 WO2017006031A1 PCT/FR2016/051670 FR2016051670W WO2017006031A1 WO 2017006031 A1 WO2017006031 A1 WO 2017006031A1 FR 2016051670 W FR2016051670 W FR 2016051670W WO 2017006031 A1 WO2017006031 A1 WO 2017006031A1
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- microorganisms
- fatty acids
- hydroxide
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- 244000005700 microbiome Species 0.000 title claims abstract description 29
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract description 26
- 239000000194 fatty acid Substances 0.000 title claims abstract description 26
- 229930195729 fatty acid Natural products 0.000 title claims abstract description 26
- 150000004665 fatty acids Chemical class 0.000 title claims abstract description 26
- 238000004519 manufacturing process Methods 0.000 title abstract description 3
- 150000002632 lipids Chemical class 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 claims abstract description 8
- 239000007864 aqueous solution Substances 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 24
- 239000007791 liquid phase Substances 0.000 claims description 16
- 238000002803 maceration Methods 0.000 claims description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 9
- -1 amine hydroxide Chemical class 0.000 claims description 9
- 150000001412 amines Chemical group 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- 230000020477 pH reduction Effects 0.000 claims description 5
- HMBHAQMOBKLWRX-UHFFFAOYSA-N 2,3-dihydro-1,4-benzodioxine-3-carboxylic acid Chemical compound C1=CC=C2OC(C(=O)O)COC2=C1 HMBHAQMOBKLWRX-UHFFFAOYSA-N 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 claims description 4
- 241000235015 Yarrowia lipolytica Species 0.000 claims description 4
- 229940075419 choline hydroxide Drugs 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- 239000007790 solid phase Substances 0.000 claims description 4
- 241000233866 Fungi Species 0.000 claims description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 3
- 241000223253 Rhodotorula glutinis Species 0.000 claims description 3
- 239000011591 potassium Substances 0.000 claims description 3
- 229910052700 potassium Inorganic materials 0.000 claims description 3
- 239000011575 calcium Substances 0.000 claims description 2
- 229940073455 tetraethylammonium hydroxide Drugs 0.000 claims description 2
- LRGJRHZIDJQFCL-UHFFFAOYSA-M tetraethylazanium;hydroxide Chemical group [OH-].CC[N+](CC)(CC)CC LRGJRHZIDJQFCL-UHFFFAOYSA-M 0.000 claims description 2
- LPSKDVINWQNWFE-UHFFFAOYSA-M tetrapropylazanium;hydroxide Chemical compound [OH-].CCC[N+](CCC)(CCC)CCC LPSKDVINWQNWFE-UHFFFAOYSA-M 0.000 claims description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 1
- 229910052791 calcium Inorganic materials 0.000 claims 1
- 159000000000 sodium salts Chemical class 0.000 claims 1
- 125000003277 amino group Chemical group 0.000 abstract 1
- 230000002879 macerating effect Effects 0.000 abstract 1
- 235000021588 free fatty acids Nutrition 0.000 description 14
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 9
- 150000003839 salts Chemical class 0.000 description 6
- 239000002028 Biomass Substances 0.000 description 5
- 239000004381 Choline salt Substances 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 239000002551 biofuel Substances 0.000 description 3
- 229960001231 choline Drugs 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 150000003626 triacylglycerols Chemical class 0.000 description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 150000001335 aliphatic alkanes Chemical class 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000003350 kerosene Substances 0.000 description 2
- 150000004702 methyl esters Chemical class 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000009482 thermal adhesion granulation Methods 0.000 description 2
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 240000009108 Chlorella vulgaris Species 0.000 description 1
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001149691 Lipomyces starkeyi Species 0.000 description 1
- 241000306281 Mucor ambiguus Species 0.000 description 1
- 241000159660 Nannochloropsis oculata Species 0.000 description 1
- 241000206744 Phaeodactylum tricornutum Species 0.000 description 1
- 241001465752 Purpureocillium lilacinum Species 0.000 description 1
- 240000005384 Rhizopus oryzae Species 0.000 description 1
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 1
- 241001524101 Rhodococcus opacus Species 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- OGBUMNBNEWYMNJ-UHFFFAOYSA-N batilol Chemical class CCCCCCCCCCCCCCCCCCOCC(O)CO OGBUMNBNEWYMNJ-UHFFFAOYSA-N 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—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
- C12P7/6409—Fatty acids
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- 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
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- 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
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/002—Sources of fatty acids, e.g. natural glycerides, characterised by the nature, the quantities or the distribution of said acids
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- 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
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/02—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils
- C11C1/025—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by saponification and release of fatty acids
Definitions
- Microorganisms can accumulate up to 80% of lipids (monoglyceride, diglyceride, triglyceride, phospholipids and sterols) in their organisms. They are composed of a cell wall, consisting mainly of lipids, polysaccharides and chitin. These compounds make the wall rigid and strong; which requires mechanical, chemical or physico-chemical pretreatment to facilitate accessibility to the lipids located inside the cell.
- the fatty acids are separated from the acidified liquid phase, for example by filtration.
- the quaternary amine contained in the liquid phase is regenerated to quaternary amine hydroxide and is recycled.
- Unconverted lipids are in the form of triglycerides (TAGs) and free fatty acids (FFAs).
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Fats And Perfumes (AREA)
Abstract
The invention relates to a method for producing fatty acids from microorganisms, comprising a step of macerating the microorganisms in which the microorganisms are mixed with an aqueous solution of a quaternary amine hydroxide in order to extract the lipids contained in the microorganisms and transform them into fatty acids.
Description
Procédé d'obtention d'acides gras à partir de microorganismes Process for obtaining fatty acids from microorganisms
L'invention a pour objet un procédé d'extraction, de transformation et de séparation des acides gras en une étape et sans l'usage de solvant organique à partir de microorganismes. The invention relates to a process for extracting, transforming and separating fatty acids in one step and without the use of organic solvent from microorganisms.
Pour la fabrication de biocarburant (en particulier le biojetfuel), on transforme les acides gras soit en alcanes (bio-kérosène) ou bien en esters méthyliques (biocarburant). Les acides gras sont obtenus à partir de l'hydrolyse de lipides préalablement extraits à partir de microorganismes (levures en particulier). Ces procédés requièrent plusieurs étapes de séchage de la biomasse, d'extraction des lipides à partir de la biomasse séchée avec un solvant organique, en général de l'hexane, puis de l'hydrolyse acide ou basique des lipides extraits en acides gras pour une transformation ultérieure en biokérosène. Les microorganismes (levures, micro-algues et bactéries) peuvent accumuler jusqu'à 80% de lipides (Monoglycéride, diglycéride, triglycéride, phospholipides et stérols) dans leurs organismes. Ils sont composés d'une paroi cellulaire, constituée principalement de lipides, de polysaccharides et de chitine. Ces composés rendent la paroi rigide et solide ; ce qui nécessite un prétraitement mécanique, chimique ou physico-chimique pour faciliter l'accessibilité aux lipides situés à l'intérieur de la cellule. For the production of biofuel (in particular the biojetfuel), the fatty acids are converted either into alkanes (bio-kerosene) or into methyl esters (biofuel). The fatty acids are obtained from the hydrolysis of lipids previously extracted from microorganisms (yeasts in particular). These processes require several steps of drying the biomass, extracting lipids from the dried biomass with an organic solvent, usually hexane, and then acid or basic hydrolysis of lipids extracted into fatty acids for further processing into biokerosene. Microorganisms (yeast, micro-algae and bacteria) can accumulate up to 80% of lipids (monoglyceride, diglyceride, triglyceride, phospholipids and sterols) in their organisms. They are composed of a cell wall, consisting mainly of lipids, polysaccharides and chitin. These compounds make the wall rigid and strong; which requires mechanical, chemical or physico-chemical pretreatment to facilitate accessibility to the lipids located inside the cell.
Actuellement, l'extraction des lipides à partir de micro-organismes en voie humide se déroule de la manière suivante : Currently, the extraction of lipids from wet microorganisms takes place as follows:
1- séchage et prétraitement de la biomasse 1 - drying and pre-treatment of biomass
2- extraction par solvants organiques (hexane) 2- extraction with organic solvents (hexane)
3- centrifugation ou filtration (recyclage du solvant) 3- centrifugation or filtration (solvent recycling)
4- Evaporation du solvant 4- Evaporation of the solvent
L'étape de séchage est nécessaire pour s'affranchir des interactions eau/solvant ; ce qui fait de l'eau un paramètre limitant dans l'extraction des lipides. L'étape de séchage est considérée comme la plus énergivore et la plus onéreuse. The drying step is necessary to overcome the water / solvent interactions; which makes water a limiting parameter in lipid extraction. The drying stage is considered the most energy consuming and the most expensive.
De plus, ce procédé utilise des solvants organiques qui sont susceptibles à moyen terme d'être interdits pour leur toxicité envers l'homme et l'environnement.
Π existe donc un besoin pour un procédé d'obtention d'acides gras à partir de microorganismes moins énergivore et plus respectueux de l'environnement. In addition, this process uses organic solvents that are likely in the medium term to be banned for their toxicity to humans and the environment. There is therefore a need for a process for obtaining fatty acids from microorganisms that consumes less energy and is more respectful of the environment.
Les inventeurs ont le mérite d'avoir mis au point un tel procédé. The inventors have the merit of having developed such a method.
La présente invention décrit un procédé d'obtention d'acides gras à partir de microorganismes comprenant une étape a) de macération des microorganismes dans laquelle les microorganismes sont mélangés à une solution aqueuse d'un hydroxyde d'amine quaternaire afin d'extraire les lipides contenus dans les microorganismes et de les transformer en acides gras. The present invention describes a process for obtaining fatty acids from microorganisms comprising a microorganism maceration step a) in which the microorganisms are mixed with an aqueous solution of a quaternary amine hydroxide to extract the lipids. contained in the microorganisms and transform them into fatty acids.
Typiquement les hydroxydes d'amine quaternaire utilisables pour cette étape de macération sont par exemple l'hydroxyde de tétraéthylammonium, l'hydroxyde de tétrapropylammonium, ou l'hydroxyde de choline. Typically, the quaternary amine hydroxides that can be used for this maceration step are, for example, tetraethylammonium hydroxide, tetrapropylammonium hydroxide, or choline hydroxide.
Selon un mode de réalisation préféré, l'hydroxyde d'amine utilisé est l'hydroxyde de choline. According to a preferred embodiment, the amine hydroxide used is choline hydroxide.
Typiquement, les microorganismes utilisables pour l'invention sont des microorganismes riches en lipides tels que les levures, les microalgues, les bactéries ou les champignons filamenteux. Typically, the microorganisms that can be used for the invention are lipid-rich microorganisms such as yeasts, microalgae, bacteria or filamentous fungi.
Typiquement, on peut citer parmi les microorganismes particulièrement adaptés pour l'invention, Rhodotorula glutinis, Yarrowia lipolytica, Lipomyces starkeyi parmi les levures, Phaeodactylum tricornutum Chlorella vulgaris, Nannochloropsis oculata parmi les microalgues, Rhodococcus opacus, Streptomyces sp, Escherichia coli parmi les bactéries, et Rhizopus arrhizus, Mucor circinelloides, pénicillium lilacinum parmi les champignons filamenteux. Typically, there may be mentioned among the microorganisms particularly suitable for the invention, Rhodotorula glutinis, Yarrowia lipolytica, Lipomyces starkeyi among yeasts, Phaeodactylum tricornutum Chlorella vulgaris, Nannochloropsis oculata among microalgae, Rhodococcus opacus, Streptomyces sp, Escherichia coli among bacteria, and Rhizopus arrhizus, Mucor circinelloides, penicillium lilacinum among filamentous fungi.
Selon un mode de réalisation préféré, les microorganismes utilisés sont des levures et en particulier une levure de l'espèce Yarrowia lipolytica ou Rhodotorula glutinis. According to a preferred embodiment, the microorganisms used are yeasts and in particular a yeast of the species Yarrowia lipolytica or Rhodotorula glutinis.
Typiquement, la biomasse constituée des microorganismes est en phase humide. Elle provient directement du bioréacteur qui a permis de la produire ou bien elle a subi une centrifugation en sortie du bioréacteur. Typically, the biomass consisting of microorganisms is in the wet phase. It comes directly from the bioreactor that made it possible to produce it or it has been centrifuged at the outlet of the bioreactor.
La biomasse est mélangée à une solution aqueuse d'un hydroxyde d'amine, ce qui permet la déstructuration des parois cellulaires, d'extraire et de transformer les lipides (tri-glycérides, di-glycérides, mono-glycérides, phospholipide) en acides gras.
Le procédé d'obtention d'acides gras selon l'invention permet d'obtenir en une seule étape a) des acides gras sous forme de sels solubles. Typiquement, l'étape de macération est réalisée avec une solution aqueuse présentant une concentration C en hydroxyde d' amine, avec Cmin < C < Cmax, les concentrations Cmin et Cmax étant choisies indépendamment l'une de l'autre, Cmin étant choisie parmi les valeurs 5%, 10%, 20% et 30%, et Cmax étant choisie parmi les valeurs 35%, 40%, 45%, 50% et 60%, les pourcentages étant des pourcentages masse d'hydroxyde d'amine/volume de solution aqueuse, préférentiellement Cmin est égale à 30% et Cmax à 50%. Selon un mode de réalisation particulier la concentration C est d'environ 45%. The biomass is mixed with an aqueous solution of an amine hydroxide, which allows the destructuration of the cell walls, to extract and transform the lipids (tri-glycerides, di-glycerides, mono-glycerides, phospholipids) into acids fat. The process for obtaining fatty acids according to the invention makes it possible to obtain in a single step a) fatty acids in the form of soluble salts. Typically, the maceration step is carried out with an aqueous solution having a concentration C of amine hydroxide, with Cmin <C <Cmax, the concentrations Cmin and Cmax being chosen independently of one another, Cmin being chosen from the values 5%, 10%, 20% and 30%, and Cmax being selected from the values 35%, 40%, 45%, 50% and 60%, the percentages being percentages mass of amine hydroxide / volume of aqueous solution, preferably Cmin is equal to 30% and Cmax to 50%. According to a particular embodiment, the concentration C is approximately 45%.
Plus la concentration C est élevée, plus la cinétique de la réaction est élevée. The higher the concentration C, the higher the kinetics of the reaction.
Typiquement le ratio masse d'hydroxyde d' amine quaternaire sur masse de microorganismes est compris entre 20 et 70 ou 30 et 60, préférentiellement entre 35 et 50. Selon un mode de réalisation particulier le ratio est d'environ 40. Typically, the mass ratio of quaternary amine hydroxide to mass of microorganisms is between 20 and 70 or 30 and 60, preferably between 35 and 50. According to a particular embodiment, the ratio is about 40.
Typiquement, l'étape de macération est réalisée pendant une durée d, avec dmin < d < dmax les durées dmin et dmax étant choisies indépendamment l'une de l'autre, dmin étant choisie parmi les valeurs 5 min, 30 min, 60 min, 2 h, 4 h et dmax étant choisie parmi les valeurs 4 h, 8 h, 12 h, 16 h, 24 h, 1 jour, 2 jours et 7 jours, préférentiellement dmin est égale à 1 h et dmax à 2 jours. Selon un mode de réalisation particulier la durée d est d'environ 2 h. Typically, the maceration step is carried out for a time d, with dmin <d <dmax the durations dmin and dmax being chosen independently of one another, dmin being chosen from values of 5 min, 30 min, 60 min. , 2 h, 4 h and dmax being chosen from the values 4 h, 8 h, 12 h, 16 h, 24 h, 1 day, 2 days and 7 days, preferably dmin is equal to 1 h and dmax to 2 days. According to a particular embodiment, the duration d is approximately 2 h.
Typiquement, l'étape de macération est réalisée à une température T avec Tmin < T < Tmax, les températures Tmin et Tmax étant choisies indépendamment l'une de l'autre, Tmin étant choisie parmi les valeurs 20, 25, 30, 35, 40, 45, 50, 55, et 60°C et Tmax étant choisie parmi les valeurs 60, 65, 70, 75, 80, et 90°C, préférentiellement Tmin est égale à 25°C et Tmax à 80°C. Selon un mode de réalisation particulier la température T est d'environ 40°C. Typically, the maceration step is carried out at a temperature T with Tmin <T <Tmax, the temperatures Tmin and Tmax being chosen independently of one another, Tmin being chosen from values 20, 25, 30, 35, 40, 45, 50, 55, and 60 ° C and Tmax being selected from the values 60, 65, 70, 75, 80, and 90 ° C, preferably Tmin is equal to 25 ° C and Tmax to 80 ° C. According to a particular embodiment, the temperature T is approximately 40 ° C.
Plus la température T est élevée, plus la cinétique de la réaction est élevée. The higher the temperature T, the higher the kinetics of the reaction.
Suite à cette étape de macération, deux phases sont obtenues : une phase solide et une phase liquide. La phase solide contient des débris cellulaires, alors que la phase liquide contient les acides gras sous forme de sel (amine quaternaire/carboxylate) soluble dans l'eau. Typiquement ces deux phases peuvent être séparées par filtration ou par centrifugation.
Ainsi le procédé selon l'invention peut comprendre une étape optionnelle b) de séparation des phases liquides et solides obtenues à l'issue de l'étape a). Pour obtenir des acides gras sous forme d'acides gras libres (FFA), il est nécessaire d'effectuer une étape supplémentaire de précipitation des acides gras compris dans la phase liquide, soit par acidification soit par ajout de sels contenant du Na+, Ca+ etc.. Ainsi le procédé selon l'invention peut comprendre une étape optionnelle c) d'acidification ou d'ajout de sels permettant de transformer en acides gras libres (FFA) les acides gras obtenus sous forme de sels lors de l'étape a) de macération. Following this maceration step, two phases are obtained: a solid phase and a liquid phase. The solid phase contains cellular debris, while the liquid phase contains fatty acids in the form of salt (quaternary amine / carboxylate) soluble in water. Typically these two phases can be separated by filtration or centrifugation. Thus, the method according to the invention may comprise an optional step b) of separation of the liquid and solid phases obtained at the end of step a). To obtain fatty acids in the form of free fatty acids (FFA), it is necessary to carry out an additional step of precipitation of the fatty acids included in the liquid phase, either by acidification or by adding salts containing Na +, Ca +, etc. Thus, the process according to the invention may comprise an optional step c) of acidification or addition of salts making it possible to convert the fatty acids obtained in the form of salts during step a) into free fatty acids (FFA). maceration.
Après la séparation, la phase liquide est acidifiée afin de faire précipiter les acides gras. Typiquement, un acide faible est utilisé pour acidifier la phase liquide, on peut, par exemple, utiliser l'acide acétique ou l'acide citrique. After separation, the liquid phase is acidified to precipitate the fatty acids. Typically, a weak acid is used to acidify the liquid phase, for example, acetic acid or citric acid may be used.
Ensuite les acides gras sont séparés de la phase liquide acidifiée, par filtration par exemple. Then the fatty acids are separated from the acidified liquid phase, for example by filtration.
Alternativement à l'acidification de la phase liquide, on peut obtenir les acides gras sous forme d'acides gras libres (FFA) en ajoutant à la phase liquide un excès de sel de sodium, de potassium ou de calcium, tel que NaCl, KCl, ou CaC^, de sorte que les acides gras précipitent sous forme de sel de sodium, de potassium ou de calcium. Le précipité ainsi obtenu est séparé de la phase liquide, les acides gras sont ensuite obtenus par attaque acide. Alternatively to the acidification of the liquid phase, fatty acids can be obtained in the form of free fatty acids (FFA) by adding to the liquid phase an excess of sodium, potassium or calcium salt, such as NaCl, KCl , or CaC ^ , so that the fatty acids precipitate as sodium, potassium or calcium salt. The precipitate thus obtained is separated from the liquid phase, the fatty acids are then obtained by acid attack.
Avantageusement, l'aminé quaternaire contenue dans la phase liquide est régénérée en hydroxyde d' aminé quaternaire, puis est recyclée. Advantageously, the quaternary amine contained in the liquid phase is regenerated to quaternary amine hydroxide and is recycled.
Typiquement, les acides gras ainsi obtenus peuvent être ultérieurement transformés en alcanes pour obtenir du bio-kérosène, ou en ester méthylique pour obtenir du biocarburant. Typically, the fatty acids thus obtained can be subsequently converted into alkanes to obtain bio-kerosene, or methyl ester to obtain biofuel.
EXEMPLES EXAMPLES
Exemple 1 Example 1
0.2g de matière sèche de Yarrowia lipolytica a été mélangé avec lOmL de choline à 45% dans l'eau pendant 2h à 40°C. Le mélange a été par la suite centrifugé pour séparer les débris cellulaires de la phase liquide contenant le sel (FFA-Choline).
Cette solution a été lavée deux fois avec 15 mL d'hexane puis analysée par chromatographie gazeuse et par chromatographie sur couche mince haute performance. 0.2 g of Yarrowia lipolytica dry matter was mixed with 10 ml of 45% choline in water for 2 h at 40 ° C. The mixture was subsequently centrifuged to separate cell debris from the salt-containing liquid phase (FFA-Choline). This solution was washed twice with 15 mL of hexane and then analyzed by gas chromatography and by high performance thin layer chromatography.
70% des lipides ont été convertis en sels FFA-choline. Les lipides non convertis sont sous forme de triglycérides (TAG) et d'acides gras libres (FFA). 70% of the lipids have been converted into FFA-choline salts. Unconverted lipids are in the form of triglycerides (TAGs) and free fatty acids (FFAs).
Exemple 2 Example 2
L'exemple 1 a été reproduit excepté le fait que le temps de macération est de 48h. 76.77% des lipides ont été convertis en sels FFA-Choline. Les lipides non convertis sont sous forme d'acides gras libres (FFA). Example 1 has been reproduced except that the maceration time is 48h. 76.77% of the lipids were converted into FFA-Choline salts. Unconverted lipids are in the form of free fatty acids (FFA).
Exemple 3 Example 3
L'exemple 1 a été reproduit excepté le fait que la température de macération est à 80°C. 74% des lipides ont été convertis en sels FFA-Choline. Les lipides non convertis sont sous forme d'acides gras libres (FFA). Example 1 was repeated except that the maceration temperature is 80 ° C. 74% of the lipids were converted into FFA-Choline salts. Unconverted lipids are in the form of free fatty acids (FFA).
Exemple 4 Example 4
L'exemple 1 a été reproduit excepté le fait que la concentration en choline est de 5%. 65% des lipides ont été convertis en sels FFA-Choline. Les lipides non convertis sont sous forme de triglycérides (TAG) et d'acides gras libres (FFA).
Example 1 was repeated except that the choline concentration is 5%. 65% of the lipids were converted into FFA-Choline salts. Unconverted lipids are in the form of triglycerides (TAGs) and free fatty acids (FFAs).
Claims
1. Procédé d'obtention d'acides gras à partir de microorganismes comprenant une étape de macération des microorganismes dans laquelle les microorganismes sont mélangés à une solution aqueuse d'un hydroxyde d' aminé quaternaire afin d'extraire les lipides contenus dans les microorganismes et de les transformer en acides gras. A process for obtaining fatty acids from microorganisms comprising a microorganism maceration step in which the microorganisms are mixed with an aqueous solution of a quaternary amine hydroxide in order to extract the lipids contained in the microorganisms and to turn them into fatty acids.
2. Procédé selon la revendication 1, dans lequel l'hydroxyde d'amine quaternaire est l'hydroxyde de tétraéthylammonium, l'hydroxyde de tétrapropylammonium, ou l'hydroxyde de choline. The process of claim 1, wherein the quaternary amine hydroxide is tetraethylammonium hydroxide, tetrapropylammonium hydroxide, or choline hydroxide.
3. Procédé selon la revendication 1, dans lequel l'hydroxyde d'amine quaternaire est l'hydroxyde de choline. The process of claim 1, wherein the quaternary amine hydroxide is choline hydroxide.
4. Procédé selon l'une quelconque des revendications 1 à 3, dans lequel les microorganismes sont des levures, des microalgues, des bactéries ou des champignons filamenteux. 4. A process according to any one of claims 1 to 3, wherein the microorganisms are yeasts, microalgae, bacteria or filamentous fungi.
5. Procédé selon la revendication 4, dans lequel les microorganismes sont des levures. 5. The method of claim 4, wherein the microorganisms are yeasts.
6. Procédé selon la revendication 5, dans lequel les microorganismes sont des levures de l'espèce Yarrowia lipolytica ou Rhodotorula glutinis. 6. The method of claim 5, wherein the microorganisms are yeasts of the species Yarrowia lipolytica or Rhodotorula glutinis.
7. Procédé selon l'une quelconque des revendications 1 à 6, dans lequel l'étape de macération est réalisée avec une solution aqueuse présentant une concentration C en hydroxyde d'amine, avec 30% < C < 50%, les pourcentages étant des pourcentages masse/volume. 7. Process according to any one of claims 1 to 6, in which the maceration step is carried out with an aqueous solution having a concentration C of amine hydroxide, with 30% <C <50%, the percentages being mass / volume percentages.
8. Procédé selon l'une quelconque des revendications 1 à 7, dans lequel le ratio masse d'hydroxyde d'amine quaternaire sur masse de microorganismes est compris entre 30 et 60. 8. Process according to any one of claims 1 to 7, wherein the mass ratio of quaternary amine hydroxide mass of microorganisms is between 30 and 60.
9. Procédé selon l'une quelconque des revendications 1 à 8, dans lequel l'étape de macération est réalisée à une température T avec 25°C < T < 80°C. 9. A process according to any one of claims 1 to 8, wherein the maceration step is carried out at a temperature T with 25 ° C <T <80 ° C.
10. Procédé selon l'une quelconque des revendications 1 à 9, dans lequel la phase liquide obtenue à l'issue de l'étape de macération est séparée de la phase solide.
10. Process according to any one of claims 1 to 9, wherein the liquid phase obtained at the end of the maceration step is separated from the solid phase.
11. Procédé selon la revendication 10, comprenant une étape de précipitation des acides gras contenus dans la phase liquide. 11. The method of claim 10, comprising a step of precipitation of fatty acids contained in the liquid phase.
12. Procédé selon la revendication 11, dans lequel la précipitation est obtenue par acidification de la phase liquide. 12. The method of claim 11, wherein the precipitation is obtained by acidification of the liquid phase.
13. Procédé selon la revendication 12, dans lequel l'acidification est réalisée avec de l'acide acétique ou de l'acide citrique. 13. The method of claim 12, wherein the acidification is carried out with acetic acid or citric acid.
14. Procédé selon la revendication 11, dans lequel la précipitation est obtenue par ajout à la phase liquide un excès de sel de sodium, de potassium ou de calcium.
14. The method of claim 11, wherein the precipitation is obtained by adding to the liquid phase an excess of sodium salt, potassium or calcium.
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| FR1556456 | 2015-07-08 | ||
| FR1556456A FR3038619A1 (en) | 2015-07-08 | 2015-07-08 | PROCESS FOR OBTAINING FATTY ACIDS FROM MICROORGANISMS |
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| PCT/FR2016/051670 WO2017006031A1 (en) | 2015-07-08 | 2016-07-01 | Method for producing fatty acids from microorganisms |
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| WO (1) | WO2017006031A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020053118A1 (en) * | 2018-09-10 | 2020-03-19 | Eni S.P.A. | Method for the extraction of bio-oil from algal biomass |
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| WO1997036996A2 (en) * | 1996-03-28 | 1997-10-09 | Gist-Brocades B.V. | Process for the preparation of a granular microbial biomass and isolation of valuable compounds therefrom |
| WO2009085801A1 (en) * | 2007-12-21 | 2009-07-09 | Old Dominion University Research Foundation | Direct conversion of biomass to biodiesel fuel |
| WO2010000416A1 (en) * | 2008-06-30 | 2010-01-07 | Eni S.P.A. | Process for the extraction of fatty acids from algal biomass |
| WO2012015635A2 (en) * | 2010-07-28 | 2012-02-02 | Uop Llc | Methods for producing phase stable, reduced acid biomass-derived pyrolysis oils |
| WO2013022963A2 (en) * | 2011-08-08 | 2013-02-14 | Old Dominion University Research Foundation | Production and separation of glycerol-related products using various feed stocks |
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2015
- 2015-07-08 FR FR1556456A patent/FR3038619A1/en not_active Ceased
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2016
- 2016-07-01 WO PCT/FR2016/051670 patent/WO2017006031A1/en active Application Filing
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO1997036996A2 (en) * | 1996-03-28 | 1997-10-09 | Gist-Brocades B.V. | Process for the preparation of a granular microbial biomass and isolation of valuable compounds therefrom |
| WO2009085801A1 (en) * | 2007-12-21 | 2009-07-09 | Old Dominion University Research Foundation | Direct conversion of biomass to biodiesel fuel |
| WO2010000416A1 (en) * | 2008-06-30 | 2010-01-07 | Eni S.P.A. | Process for the extraction of fatty acids from algal biomass |
| WO2012015635A2 (en) * | 2010-07-28 | 2012-02-02 | Uop Llc | Methods for producing phase stable, reduced acid biomass-derived pyrolysis oils |
| WO2013022963A2 (en) * | 2011-08-08 | 2013-02-14 | Old Dominion University Research Foundation | Production and separation of glycerol-related products using various feed stocks |
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| MILANESIO J ET AL: "Extraction of lipids from Yarrowia Lipolytica", JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, WILEY, vol. 88, no. 3, 1 March 2013 (2013-03-01), pages 378 - 387, XP002733695, ISSN: 0268-2575, [retrieved on 20120612], DOI: 10.1002/JCTB.3840 * |
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| WO2020053118A1 (en) * | 2018-09-10 | 2020-03-19 | Eni S.P.A. | Method for the extraction of bio-oil from algal biomass |
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| FR3038619A1 (en) | 2017-01-13 |
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