WO2008155410A1 - Production of lipids containing poly-unsaturated fatty acids - Google Patents

Production of lipids containing poly-unsaturated fatty acids Download PDF

Info

Publication number
WO2008155410A1
WO2008155410A1 PCT/EP2008/057866 EP2008057866W WO2008155410A1 WO 2008155410 A1 WO2008155410 A1 WO 2008155410A1 EP 2008057866 W EP2008057866 W EP 2008057866W WO 2008155410 A1 WO2008155410 A1 WO 2008155410A1
Authority
WO
WIPO (PCT)
Prior art keywords
lipid
acid
separation
emulsion
pufa
Prior art date
Application number
PCT/EP2008/057866
Other languages
French (fr)
Inventor
Per Munk Nielsen
Mogens WÜMPELMANN
Original Assignee
Novozymes A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novozymes A/S filed Critical Novozymes A/S
Publication of WO2008155410A1 publication Critical patent/WO2008155410A1/en

Links

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
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Definitions

  • the invention relates to recovery of lipids containing poly-unsatu rated fatty acids.
  • lipids containing polyunsaturated fatty acids such as omega-3 (n-3) oils as a healthy food ingredient
  • PUFA polyunsaturated fatty acids
  • Such lipids may be produced by cultivation of algal cells, followed by cell cracking (or lysis) (WO 01/53512, WO 01/54510, US 2006/0286205). It is known that after the cell lysis, quality and stability of the poly-unsaturated lipid may deteriorate due to oxidation of the unsaturated olefinic bonds. It is further known that metallic ions may catalyze the oxidative reaction, and chelators may be added to reduce the speed of the oxidative damage.
  • WO 01/53512 (Omegatech) describes a method for extracting lipids from microor- ganisms by lysing cells and removing water soluble materials by washing until a non- emulsified lipid is obtained.
  • US 2006/0286205 (to Martek) describes methods for producing a product comprising a long chain polyunsaturated fatty acid by hydrolyzing biomass and emulsifying the hy- drolyzed biomass.
  • US 2004/0170728 (to Kraft) describes the use of antioxidant compositions containing a siderophore such as rhodotorulic acid and an organic acid such as lactobionic acid to retard oxidation of lipids such as fish oil.
  • the invention provides an improved process for recovery of lipids containing poly- unsaturated fatty acids (PUFA) from a culture broth of algae with intracellular PUFA lipids.
  • PUFA poly- unsaturated fatty acids
  • the invention leads to a PUFA product with a low level of free or total metal ions (particularly iron and copper ions) and good oxidation stability, and this can be achieved in a simple process with few separation steps without the need for a siderophore. Fewer separation steps may lead to less loss of antioxidant and hence a lower need for addition of antioxidant to achieve good oxidation stability.
  • the invention provides a process for producing a lipid, comprising: a) cultivating microbial cells so as to produce intracellular lipid comprising a polyunsaturated fatty acyl group, b) lysing the microbial cells in the culture broth so as to release the lipid, c) separating the lysate by gravitational force to collect a fraction which is a lipid- containing emulsion, d) adding a metal chelating agent during or after the lysis or the separation.
  • Lipids containing poly-unsaturated fatty acid PUFA
  • the PUFA-containing lipids produced in the process of the invention may comprise mono-, di- or triglycerides comprising a poly-unsaturated fatty acyl group.
  • the polyunsaturated fatty acids may include omega-3 or omega-6 fatty acid such as docosa- 4:7:10:13:16:19-hexaenoic acid (DHA), eicosapentaenoic acid (EPA) and docosapentaenoic acid (DPA).
  • the microbial cells may be Eukaryota; stramenopiles; Laby ⁇ nthulida;
  • Thraustoch ytriidae Schizochytrium or Thraustochytrium, e.g. S. limacinum. They may be fungi or algae.
  • the microbial cells are cultivated so as to produce intracellular lipid. This may be done as described in EP 823475 (Agency of Industrial Science and Technology et al.), JP
  • the microorganism may form structured triglycerides mainly with one poly- unsaturated fatty acyl group in the sn2 position and saturated or mono-unsaturated acyl (e.g. palmitoyl) in the sn1 and/or the sn3 positions, or it may form an unstructured triglyceride with an essentially random distribution.
  • structured triglycerides mainly with one poly- unsaturated fatty acyl group in the sn2 position and saturated or mono-unsaturated acyl (e.g. palmitoyl) in the sn1 and/or the sn3 positions, or it may form an unstructured triglyceride with an essentially random distribution.
  • Lysis of microbial cells The culture broth after cultivation is treated so as to lyse the microbial cells and release the lipid. This may be done by bead milling, homogenization, treatment with a cell-wall degrading enzyme, addition of an alkali or heating.
  • the cell-wall degrading enzyme may include protease, alginate lyase, pectinase, phospholipase or a carbohydrate hydrolase such as a cellulase, hemicellulase.
  • Homogenization is advantageously done at high temperature (above 45 0 C) and high pressure (e.g., 500-1000 bars). Two or more runs through the homogenizer may be used in order to break a large percentage of the cells.
  • the separation by gravitational force conveniently includes centrifugation where the lipid-containing emulsion is collected as the top phase. Continuous centrifugation may be advantageous.
  • the separation may include one or more additional cycles, where each cycle includes adding water to the lipid emulsion, agitating, and separating by centrifugation.
  • the emulsion collected after the first centrifugation may particularly be an oil-in- water emulsion.
  • the emulsion collected after the last cycle may be a water-in-oil emulsion and/or may have a water content of 0.1-10 %, particularly 1-10 % by weight.
  • the metal chelator may include EDTA (ethylene-diamine tetra-acetic acid), an aldo- bionic acid (such as lactobionic acid), phosphoric acid, gluconic acid, citric acid, transferrin
  • EDTA ethylene-diamine tetra-acetic acid
  • aldo- bionic acid such as lactobionic acid
  • phosphoric acid phosphoric acid
  • gluconic acid gluconic acid
  • citric acid transferrin
  • the metal chelator may be added up to as much as can nutritionally be responsible to add, e.g. in an amount of 10-10,000 ppm by weight, particularly 100-5,000 ppm.
  • the oxygen scavenger may be added in addition to the metal chelator.
  • the oxygen scavenger may comprise laccase, superoxide dismutase or an enzymatic system comprising glucose oxidase, catalase and glucose, e.g. as described in WO9631 133, US3920521 -A, EP649603 or EP934702.
  • the oxygen scavenger may comprise an anti-oxidant such as cocoa, tocopherol or resveratrol, e.g. in an amount of 0.1-1 % by weight.
  • the hydrolysis may advantageously be done by incubation with a lipase which preferentially hydrolyzes saturated fatty acids rather than polyunsaturated fatty acids, e.g. the lipase from Candida rugosa.
  • a lipase which preferentially hydrolyzes saturated fatty acids rather than polyunsaturated fatty acids
  • the hydrolysis may advanta- geously be done with a 1 ,3-specific lipase so as to hydrolyze saturated fatty acids in the sn1 and sn3 positions and thereby increase the PUFA content of the resulting lipid.
  • the 1 ,3- specific lipase may be derived from Thermomyces lanuginosus (EP305216), Rhizomucor miehei, Aspergillus niger, Mucorjavanicus, Rhizopus delemar, or Rhizopus arrhizus.
  • the free fatty acid may be removed by converting it to a methyl ester and distillation. Oxidation stability
  • the oxidative stability of PUFA lipid can be evaluated by determination of peroxide value, e.g. by AOCS Official method Cd 8b-90 (Peroxide Value, Acetic Acid - lsooctane Method) or as described in R. Abuzaytoun et al., J. Agric. Food Chem., 54 (21 ), 8253 -8260, 2006.
  • Example 1 Recovery of PUFA-containing lipid from algal culture broth
  • PUFA-containing lipid was produced and recovered by the following process:
  • lipids (by weight) of the mature cells was lipids. 25-30 % of the lipid content was DHA (22:6(n-3)), and 50-60 % was palmitic acid (16:0).
  • the lipid in the algae was present mainly as lipid vesicles with triglycerides (3 fatty acids pr molecule), and as sterol esters (1 fatty acid) and phospholipids (2 fatty acids) in the cell and organelle membranes.
  • a 750 L pressure tank was heated to 75-8O 0 C with steam and flushed with nitrogen.
  • the pressure was kept at 1 atm + 0.2 bars after flushing.
  • the culture broth was run through a bead mill with 0.6 mm SiLi beads (Zirconium Oxide), Flow was 200 L/h, after which the fluid was heated in a 1 -2 m 2 heat exchanger to 75 0 C, and transferred to the pressure tank.
  • the pH in the pressure tank was regulated to pH 10 with 45% KOH, as soon as the fluid level reached the pH sensor in the tank.
  • Pre separation A Westfalia SB-7 centrifuge was heated to 75 0 C with steam, and jerrycans were prepared for product reception by nitrogen-flushing.
  • the alkaline liquid still over 7O 0 C was run through the SB-7 oil centrifuge (200 L/h, 4.0-4.4 bars counter pressure), and the lipid emul- sion was recovered as the top phase.
  • the lipid emulsion was collected in 25 L jerrycans.

Abstract

The invention provides an improved process for recovery of lipids containing polyunsaturated fatty acids (PUFA) from a culture broth of algae with intracellular PUFA lipids. The process of the invention allows production of a PUFA lipid product with high oxidation stability at a low production cost. Accordingly, the invention provides a process for producing a lipid, comprising: a) cultivating microbial cells so as to produce intracellular lipid comprising a polyunsaturated fatty acyl group, b) lysing the microbial cells in the culture broth so as to release the lipid, c) separating the lysate by gravitational force to collect a fraction which is a lipid containing emulsion, d) adding a metal chelating agent during or after the lysis or the separation.

Description

PRODUCTION OF LIPIDS CONTAINING POLY-U N SAT U RATE D FATTY ACIDS
FIELD OF THE INVENTION
The invention relates to recovery of lipids containing poly-unsatu rated fatty acids.
BACKGROUND OF THE INVENTION The worldwide market for lipids containing polyunsaturated fatty acids (PUFA) such as omega-3 (n-3) oils as a healthy food ingredient is increasing rapidly, in the areas of adult supplements, infant formula and as food ingredients. Such lipids may be produced by cultivation of algal cells, followed by cell cracking (or lysis) (WO 01/53512, WO 01/54510, US 2006/0286205). It is known that after the cell lysis, quality and stability of the poly-unsaturated lipid may deteriorate due to oxidation of the unsaturated olefinic bonds. It is further known that metallic ions may catalyze the oxidative reaction, and chelators may be added to reduce the speed of the oxidative damage.
WO 01/53512 (Omegatech) describes a method for extracting lipids from microor- ganisms by lysing cells and removing water soluble materials by washing until a non- emulsified lipid is obtained.
US 2006/0286205 (to Martek) describes methods for producing a product comprising a long chain polyunsaturated fatty acid by hydrolyzing biomass and emulsifying the hy- drolyzed biomass. US 2004/0170728 (to Kraft) describes the use of antioxidant compositions containing a siderophore such as rhodotorulic acid and an organic acid such as lactobionic acid to retard oxidation of lipids such as fish oil.
SUMMARY OF THE INVENTION
The invention provides an improved process for recovery of lipids containing poly- unsaturated fatty acids (PUFA) from a culture broth of algae with intracellular PUFA lipids.
More particularly, the invention leads to a PUFA product with a low level of free or total metal ions (particularly iron and copper ions) and good oxidation stability, and this can be achieved in a simple process with few separation steps without the need for a siderophore. Fewer separation steps may lead to less loss of antioxidant and hence a lower need for addition of antioxidant to achieve good oxidation stability.
Accordingly, the invention provides a process for producing a lipid, comprising: a) cultivating microbial cells so as to produce intracellular lipid comprising a polyunsaturated fatty acyl group, b) lysing the microbial cells in the culture broth so as to release the lipid, c) separating the lysate by gravitational force to collect a fraction which is a lipid- containing emulsion, d) adding a metal chelating agent during or after the lysis or the separation..
DETAILED DESCRIPTION OF THE INVENTION
Lipids containing poly-unsaturated fatty acid (PUFA)
The PUFA-containing lipids produced in the process of the invention may comprise mono-, di- or triglycerides comprising a poly-unsaturated fatty acyl group. The polyunsaturated fatty acids may include omega-3 or omega-6 fatty acid such as docosa- 4:7:10:13:16:19-hexaenoic acid (DHA), eicosapentaenoic acid (EPA) and docosapentaenoic acid (DPA).
Cultivation of microbial cells
The microbial cells may be Eukaryota; stramenopiles; Labyήnthulida;
Thraustoch ytriidae; Schizochytrium or Thraustochytrium, e.g. S. limacinum. They may be fungi or algae. The microbial cells are cultivated so as to produce intracellular lipid. This may be done as described in EP 823475 (Agency of Industrial Science and Technology et al.), JP
1 1-318484A (Sagami Chem. Res. Cent, et al.), WO 01/53512 (Omegatech), US
2006/0286205 (to Martek), and WO 01/54510 (Omegatech).
The microorganism may form structured triglycerides mainly with one poly- unsaturated fatty acyl group in the sn2 position and saturated or mono-unsaturated acyl (e.g. palmitoyl) in the sn1 and/or the sn3 positions, or it may form an unstructured triglyceride with an essentially random distribution.
Lysis of microbial cells The culture broth after cultivation is treated so as to lyse the microbial cells and release the lipid. This may be done by bead milling, homogenization, treatment with a cell-wall degrading enzyme, addition of an alkali or heating.
The cell-wall degrading enzyme may include protease, alginate lyase, pectinase, phospholipase or a carbohydrate hydrolase such as a cellulase, hemicellulase. Homogenization is advantageously done at high temperature (above 450C) and high pressure (e.g., 500-1000 bars). Two or more runs through the homogenizer may be used in order to break a large percentage of the cells. Gravitational separation
The separation by gravitational force conveniently includes centrifugation where the lipid-containing emulsion is collected as the top phase. Continuous centrifugation may be advantageous. The separation may include one or more additional cycles, where each cycle includes adding water to the lipid emulsion, agitating, and separating by centrifugation.
The emulsion collected after the first centrifugation may particularly be an oil-in- water emulsion. The emulsion collected after the last cycle may be a water-in-oil emulsion and/or may have a water content of 0.1-10 %, particularly 1-10 % by weight.
Metal-chelating agent
The metal chelator may include EDTA (ethylene-diamine tetra-acetic acid), an aldo- bionic acid (such as lactobionic acid), phosphoric acid, gluconic acid, citric acid, transferrin
(e.g. produced recombinantly or isolated from milk) or another protein chelator. The metal chelator may be added up to as much as can nutritionally be responsible to add, e.g. in an amount of 10-10,000 ppm by weight, particularly 100-5,000 ppm.
Optional oxygen scavenger
An oxygen scavenger may be added in addition to the metal chelator. The oxygen scavenger may comprise laccase, superoxide dismutase or an enzymatic system comprising glucose oxidase, catalase and glucose, e.g. as described in WO9631 133, US3920521 -A, EP649603 or EP934702. Alternatively, the oxygen scavenger may comprise an anti-oxidant such as cocoa, tocopherol or resveratrol, e.g. in an amount of 0.1-1 % by weight.
Optional triglyceride hydrolysis
It may be desirable to include hydrolysis of triglycerides to diglycerides with a higher PUFA content and free fatty acids with a lower PUFA content, followed by separation to re- move free fatty acid. This may be done after the lipid separation and before the chelator addition.
The hydrolysis may advantageously be done by incubation with a lipase which preferentially hydrolyzes saturated fatty acids rather than polyunsaturated fatty acids, e.g. the lipase from Candida rugosa. In the case of structured lipid, the hydrolysis may advanta- geously be done with a 1 ,3-specific lipase so as to hydrolyze saturated fatty acids in the sn1 and sn3 positions and thereby increase the PUFA content of the resulting lipid. The 1 ,3- specific lipase may be derived from Thermomyces lanuginosus (EP305216), Rhizomucor miehei, Aspergillus niger, Mucorjavanicus, Rhizopus delemar, or Rhizopus arrhizus.
The free fatty acid may be removed by converting it to a methyl ester and distillation. Oxidation stability
The oxidative stability of PUFA lipid can be evaluated by determination of peroxide value, e.g. by AOCS Official method Cd 8b-90 (Peroxide Value, Acetic Acid - lsooctane Method) or as described in R. Abuzaytoun et al., J. Agric. Food Chem., 54 (21 ), 8253 -8260, 2006.
EXAMPLES
Example 1 : Recovery of PUFA-containing lipid from algal culture broth
PUFA-containing lipid was produced and recovered by the following process:
Cultivation of cells Cells of Schizochytrium sp. were cultivated to obtain a culture broth. Approx. 50 %
(by weight) of the mature cells was lipids. 25-30 % of the lipid content was DHA (22:6(n-3)), and 50-60 % was palmitic acid (16:0). The lipid in the algae was present mainly as lipid vesicles with triglycerides (3 fatty acids pr molecule), and as sterol esters (1 fatty acid) and phospholipids (2 fatty acids) in the cell and organelle membranes. Pre lysis
A 750 L pressure tank was heated to 75-8O0C with steam and flushed with nitrogen.
The pressure was kept at 1 atm + 0.2 bars after flushing.
Lysis of cells
The culture broth was run through a bead mill with 0.6 mm SiLi beads (Zirconium Oxide), Flow was 200 L/h, after which the fluid was heated in a 1 -2 m2 heat exchanger to 750C, and transferred to the pressure tank.
The pH in the pressure tank was regulated to pH 10 with 45% KOH, as soon as the fluid level reached the pH sensor in the tank.
Pre separation A Westfalia SB-7 centrifuge was heated to 750C with steam, and jerrycans were prepared for product reception by nitrogen-flushing.
Separation
After 1.5-4.0 hours in the pressure tank, the alkaline liquid still over 7O0C was run through the SB-7 oil centrifuge (200 L/h, 4.0-4.4 bars counter pressure), and the lipid emul- sion was recovered as the top phase. The lipid emulsion was collected in 25 L jerrycans.
In separate experiments, it was found that very often water should be added in order to get a good separation; 1 to 2 volumes of water to 1 volume alkaline lysed culture broth, especially if the culture broth is rich in biomass.

Claims

1. A process for producing a lipid, comprising: a) cultivating microbial cells so as to produce intracellular lipid comprising a polyunsaturated fatty acyl group, b) lysing the microbial cells in the culture broth so as to release the lipid, c) separating the lysate by gravitational force to collect a fraction which is a lipid- containing emulsion, d) adding a metal chelating agent during or after the lysis or the separation.
2. The process of claim 1 wherein the lysis comprises bead milling, homogenization, treat- ment with a cell-wall degrading enzyme, addition of an alkali or heating.
3. The process of any preceding claim wherein the separation step comprises centrifuga- tion.
4. The process of either preceding claim which further comprises at least one cycle between c) and d), wherein each cycle comprises adding water, agitation, separation by gravitational force to collect a fraction which is a lipid-containing emulsion.
5. The process of the preceding claim wherein the emulsion formed in step c) is an oil-in- water emulsion, and wherein the emulsion formed after the last cycle is a water-in-oil emulsion or has a water content of 0.1-10 % by weight.
6. The process of any preceding claim wherein the metal chelating agent comprises EDTA (ethylene-diamine tetra-acetic acid), an aldobionic acid (particularly lactobionic acid), phosphoric acid, gluconic acid, citric acid, or a protein chelator (particularly transferrin).
7. The process of any preceding claim wherein step d) further comprises adding an oxygen- scavenger.
8. The process of the preceding claim wherein the oxygen scavenger comprises laccase, superoxide dismutase or a system comprising glucose oxidase, catalase and glucose.
9. The process of a preceding claim wherein the oxygen scavenger comprises an antioxidant, particularly cocoa, tocopherol or resveratrol.
10. The process of any preceding claim which further comprises the following steps after the separation and before the addition of metal chelating agent: a) hydrolyzing triglycerides to form diglycerides and free fatty acid, followed by b) removing free fatty acids.
1 1. The process of the preceding claim wherein the hydrolysis comprises incubating with a lipase, particularly a 1 ,3-specific lipase or a lipase with preference for saturated fatty acyl groups.
PCT/EP2008/057866 2007-06-21 2008-06-20 Production of lipids containing poly-unsaturated fatty acids WO2008155410A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP07110805 2007-06-21
EP07110805.4 2007-06-21

Publications (1)

Publication Number Publication Date
WO2008155410A1 true WO2008155410A1 (en) 2008-12-24

Family

ID=38626555

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2008/057866 WO2008155410A1 (en) 2007-06-21 2008-06-20 Production of lipids containing poly-unsaturated fatty acids

Country Status (1)

Country Link
WO (1) WO2008155410A1 (en)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CZ302118B6 (en) * 2010-06-02 2010-10-20 Botanický ústav AV CR, v. v. i. Trachydiscus minutus algae, Lukavsky & Pribyl 2005/1 strain producing oils with high amount of higher unsaturated fatty acids
NL2004832C2 (en) * 2010-06-07 2011-12-08 Evodos B V Separating biomass from an aqueous medium.
ITMI20101666A1 (en) * 2010-09-13 2012-03-14 Dmf Dietetic Metabolic Food S R L COMPOSITION BASED ON DOCOSAESAENOIC ACID (DHA), IN THE TREATMENT OF CYSTIC FIBROSIS AND NON-ALCOHOLIC HEPATIC STEATOSIS
US20130171721A1 (en) * 2011-12-30 2013-07-04 Alliance For Sustainable Energy, Llc Disruption of cell walls for enhanced lipid recovery
WO2015017445A1 (en) * 2013-07-29 2015-02-05 The Regents Of The University Of California Fatty acid production in cell-free systems
EP2183376A4 (en) * 2007-07-12 2016-08-10 Dsm Nutritional Products Ag Enzymatic modification of oil
CZ307402B6 (en) * 2017-06-08 2018-07-25 Mikrobiologický ústav AV ČR v. v. i. A production strain of the Bracteacoccus bullatus alga for the production of oils containing essential unsaturated fatty acids, a method of producing these oils and the use of this strain for the industrial production of these oils
EP2576801B1 (en) 2010-06-01 2019-10-02 DSM IP Assets B.V. Extraction of lipid from cells and products therefrom
WO2020186130A1 (en) * 2019-03-14 2020-09-17 Dsm Ip Assets B.V. Methods of obtaining lipids from a microbial cell composition by enzyme and ph shock

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991007498A1 (en) * 1989-11-17 1991-05-30 Phycotech, Inc. Process for the heterotrophic production of products with high concentrations of omega-3 highly unsaturated fatty acids
EP0823475A1 (en) * 1995-04-17 1998-02-11 Japan as represented by Director-General, Agency of Industrial Science and Technology Novel microorganisms capable of producing highly unsaturated fatty acids and process for producing highly unsaturated fatty acids by using the microorganisms
WO2001053512A1 (en) * 2000-01-19 2001-07-26 Omegatech, Inc. Solventless extraction process
WO2001054510A1 (en) * 2000-01-28 2001-08-02 Omegatech, Inc. Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors
US20040170728A1 (en) * 2003-02-28 2004-09-02 Kraft Foods Holdings, Inc. Use of siderophores and organic acids to retard lipid oxidation
WO2005040318A1 (en) * 2003-10-21 2005-05-06 Dsm Ip Assets B.V. Stabilisation of polyunsaturated fatty acid (pufa) ester concentrates
WO2006031699A2 (en) * 2004-09-10 2006-03-23 Diversa Corporation Compositions and methods for making and modifying oils
WO2006046943A2 (en) * 2004-10-22 2006-05-04 Martek Biosciences Corporation Methods for producing lipids by liberation from biomass
US20060286205A1 (en) * 2005-05-12 2006-12-21 Martek Biosciences Corporation Biomass hydrolysate and uses and production thereof

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991007498A1 (en) * 1989-11-17 1991-05-30 Phycotech, Inc. Process for the heterotrophic production of products with high concentrations of omega-3 highly unsaturated fatty acids
EP0823475A1 (en) * 1995-04-17 1998-02-11 Japan as represented by Director-General, Agency of Industrial Science and Technology Novel microorganisms capable of producing highly unsaturated fatty acids and process for producing highly unsaturated fatty acids by using the microorganisms
WO2001053512A1 (en) * 2000-01-19 2001-07-26 Omegatech, Inc. Solventless extraction process
WO2001054510A1 (en) * 2000-01-28 2001-08-02 Omegatech, Inc. Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors
US20040170728A1 (en) * 2003-02-28 2004-09-02 Kraft Foods Holdings, Inc. Use of siderophores and organic acids to retard lipid oxidation
WO2005040318A1 (en) * 2003-10-21 2005-05-06 Dsm Ip Assets B.V. Stabilisation of polyunsaturated fatty acid (pufa) ester concentrates
WO2006031699A2 (en) * 2004-09-10 2006-03-23 Diversa Corporation Compositions and methods for making and modifying oils
WO2006046943A2 (en) * 2004-10-22 2006-05-04 Martek Biosciences Corporation Methods for producing lipids by liberation from biomass
US20060286205A1 (en) * 2005-05-12 2006-12-21 Martek Biosciences Corporation Biomass hydrolysate and uses and production thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
WARD ET AL: "Omega-3/6 fatty acids: Alternative sources of production", PROCESS BIOCHEMISTRY, vol. 40, no. 12, December 2005 (2005-12-01), pages 3627 - 3652, XP005637930, ISSN: 1359-5113 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2183376A4 (en) * 2007-07-12 2016-08-10 Dsm Nutritional Products Ag Enzymatic modification of oil
US11649473B2 (en) 2007-07-12 2023-05-16 Dsm Nutritional Products Ag Enzymatic modification of oil
US9885068B2 (en) 2007-07-12 2018-02-06 Dsm Ip Assets B.V. Enzymatic modification of oil
EP2576801B1 (en) 2010-06-01 2019-10-02 DSM IP Assets B.V. Extraction of lipid from cells and products therefrom
CZ302118B6 (en) * 2010-06-02 2010-10-20 Botanický ústav AV CR, v. v. i. Trachydiscus minutus algae, Lukavsky & Pribyl 2005/1 strain producing oils with high amount of higher unsaturated fatty acids
WO2011155828A1 (en) * 2010-06-07 2011-12-15 Evodos Algae Technologies B.V. Separating biomass from an aqueous medium
NL2004832C2 (en) * 2010-06-07 2011-12-08 Evodos B V Separating biomass from an aqueous medium.
EP2428217A1 (en) * 2010-09-13 2012-03-14 DMF Dietetic Metabolic Food S.R.L. Composition based on docosahexaenoic acid (dha) for use in the treatment of cystic fibrosis and non-alcoholic hepatic steatosis
ITMI20101666A1 (en) * 2010-09-13 2012-03-14 Dmf Dietetic Metabolic Food S R L COMPOSITION BASED ON DOCOSAESAENOIC ACID (DHA), IN THE TREATMENT OF CYSTIC FIBROSIS AND NON-ALCOHOLIC HEPATIC STEATOSIS
US20130171721A1 (en) * 2011-12-30 2013-07-04 Alliance For Sustainable Energy, Llc Disruption of cell walls for enhanced lipid recovery
US8986977B2 (en) * 2011-12-30 2015-03-24 Alliance For Sustainable Energy, Llc Disruption of cell walls for enhanced lipid recovery
WO2015017445A1 (en) * 2013-07-29 2015-02-05 The Regents Of The University Of California Fatty acid production in cell-free systems
US10155968B2 (en) 2013-07-29 2018-12-18 The Regents Of The University Of California Fatty acid production in cell-free systems
CZ307402B6 (en) * 2017-06-08 2018-07-25 Mikrobiologický ústav AV ČR v. v. i. A production strain of the Bracteacoccus bullatus alga for the production of oils containing essential unsaturated fatty acids, a method of producing these oils and the use of this strain for the industrial production of these oils
WO2020186130A1 (en) * 2019-03-14 2020-09-17 Dsm Ip Assets B.V. Methods of obtaining lipids from a microbial cell composition by enzyme and ph shock
EP3938480A4 (en) * 2019-03-14 2022-12-28 DSM IP Assets B.V. Methods of obtaining lipids from a microbial cell composition by enzyme and ph shock

Similar Documents

Publication Publication Date Title
WO2008155410A1 (en) Production of lipids containing poly-unsaturated fatty acids
US7431952B2 (en) Isolation of microbial oils
Donot et al. Single cell oils (SCOs) from oleaginous yeasts and moulds: production and genetics
Ghazani et al. Microbial lipids for foods
AU2001293711A1 (en) Isolation of microbial oils
JP2005102680A (en) Microorganism having docosahexaenoic acid producing ability and utilization of the same
JPWO2006022356A1 (en) Method for producing microbial fats and oils optionally containing diacylglycerol content and the fats and oils
WO2006046943A2 (en) Methods for producing lipids by liberation from biomass
Mamani et al. Industrial production, patent landscape, and market trends of arachidonic acid-rich oil of Mortierella alpina
Araújo et al. Synthesis of structured triacylglycerols enriched in n-3 fatty acids by immobilized microbial lipase
JP5416861B2 (en) Method for producing highly unsaturated fatty acid-containing fat with lipase
US20080248187A1 (en) Mixture containing fatty acid glycerides
Nagao et al. Purification of conjugated linoleic acid isomers through a process including lipase-catalyzed selective esterification
DE102005057832A1 (en) Process for the enzymatic synthesis of triglycerides
Mukherjee Lipid biotechnology
Fregolente et al. Screening of microbial lipases and evalutaion of their potential to produce glycerides with high gamma linolenic acid concentration
Sharma et al. Fungus‐mediated generation of ethyl ester using acid oil as substrate
Zaharudin et al. Oleic acid enhancement in used frying palm oil via enzymatic acidolysis
Weber et al. 28Lipid Biotechnology
Sancholle et al. Biotechnological potential of fungal lipids
Erdir Kolcu High-pressure methods for extraction of lipids from Rhodosporidium toruloides
Carvalho et al. Enrichment of gamma‐linolenic acid from fungal oil by lipases
Hasnisa et al. Enzymatic acidolysis of palm olein with PUFA to improve linoleic and α-linolenic acids ratio.
PARK et al. Enzymatic synthesis of ethyl ester of highly unsaturated fatty acids from fish oils using immobilized lipase
Balduyck Lipolytic stability during wet storage of autotrophic microalgae

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08761273

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08761273

Country of ref document: EP

Kind code of ref document: A1