AU622061B2 - A process for the production of polyunsaturated fatty acid - Google Patents
A process for the production of polyunsaturated fatty acid Download PDFInfo
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- AU622061B2 AU622061B2 AU68456/90A AU6845690A AU622061B2 AU 622061 B2 AU622061 B2 AU 622061B2 AU 68456/90 A AU68456/90 A AU 68456/90A AU 6845690 A AU6845690 A AU 6845690A AU 622061 B2 AU622061 B2 AU 622061B2
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- polyunsaturated fatty
- fatty acids
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- 238000000034 method Methods 0.000 title claims description 20
- 230000008569 process Effects 0.000 title claims description 19
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title claims description 18
- 238000004519 manufacturing process Methods 0.000 title claims description 9
- 239000001963 growth medium Substances 0.000 claims description 17
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 11
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 10
- 238000009630 liquid culture Methods 0.000 claims description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 8
- 241000518454 Rhytidiadelphus squarrosus Species 0.000 claims description 6
- 239000011782 vitamin Substances 0.000 claims description 5
- 229940088594 vitamin Drugs 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- SXGZJKUKBWWHRA-UHFFFAOYSA-N 2-(N-morpholiniumyl)ethanesulfonate Chemical compound [O-]S(=O)(=O)CC[NH+]1CCOCC1 SXGZJKUKBWWHRA-UHFFFAOYSA-N 0.000 claims description 4
- 241000518474 Brachythecium rutabulum Species 0.000 claims description 4
- 241001600678 Brachythecium salebrosum Species 0.000 claims description 4
- 241000184277 Eurhynchium striatum Species 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- 241001600662 Pseudoscleropodium purum Species 0.000 claims description 4
- 241001136234 Rhytidiadelphus triquetrus Species 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 4
- 239000002537 cosmetic Substances 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 239000000122 growth hormone Substances 0.000 claims description 4
- 235000013305 food Nutrition 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 25
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 12
- 239000000243 solution Substances 0.000 description 11
- 235000014113 dietary fatty acids Nutrition 0.000 description 8
- 239000000194 fatty acid Substances 0.000 description 8
- 229930195729 fatty acid Natural products 0.000 description 8
- 150000002632 lipids Chemical class 0.000 description 8
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 7
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 7
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 7
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 7
- 229940114079 arachidonic acid Drugs 0.000 description 6
- 235000021342 arachidonic acid Nutrition 0.000 description 6
- 150000004665 fatty acids Chemical class 0.000 description 6
- 241000195940 Bryophyta Species 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 238000011534 incubation Methods 0.000 description 4
- 150000004702 methyl esters Chemical class 0.000 description 4
- WETWJCDKMRHUPV-UHFFFAOYSA-N acetyl chloride Chemical compound CC(Cl)=O WETWJCDKMRHUPV-UHFFFAOYSA-N 0.000 description 3
- 239000012346 acetyl chloride Substances 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 3
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 3
- 239000002028 Biomass Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 210000000712 G cell Anatomy 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QWDCYFDDFPWISL-UHFFFAOYSA-N UNPD207407 Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(=O)OC QWDCYFDDFPWISL-UHFFFAOYSA-N 0.000 description 2
- OFIDNKMQBYGNIW-UHFFFAOYSA-N arachidonic acid methyl ester Natural products CCCCCC=CCC=CCC=CCC=CCCCC(=O)OC OFIDNKMQBYGNIW-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- QWDCYFDDFPWISL-JEBPEJKESA-N cis-5,8,11,14,17-eicosapentaenoic acid methyl ester Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)OC QWDCYFDDFPWISL-JEBPEJKESA-N 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- -1 fatty acid esters Chemical class 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- OFIDNKMQBYGNIW-ZKWNWVNESA-N methyl arachidonate Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)OC OFIDNKMQBYGNIW-ZKWNWVNESA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 101150050425 CCC2 gene Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 241001480517 Conidiobolus Species 0.000 description 1
- 241001491638 Corallina Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 241000235575 Mortierella Species 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000021121 meiosis Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 150000003180 prostaglandins Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000005809 transesterification reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 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
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
-
- 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
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6432—Eicosapentaenoic acids [EPA]
-
- 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
- C12P7/6458—Glycerides by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
-
- 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cosmetics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
e r 1_~ COMMONWEALTH OF AUSTRALIA FORM PATENTS CT 1952 C 0 MPLETE SPEC I F I CATION FOR OFFICE USE: Class Int.Class Application Number: Lodged: 0 tu Complete Specification Lodged: S. Accepted: Published: .',Priority: ."Related Art: r Name of Applicant: Address f Applicant: :Address of Applicant: SOCIETE DES PRODUITS NESTLE S.A.
Vevey, Switzerland Christian BOREL and Carl Erik HANSEN 0* Actual Inventor: SAddress for Service: SHELSTON WATERS, 55 Clarence Street, Sydney Complete Specification for the Invention entitled: "A PROCESS FOR THE PRODUCTION OF POLYUNSATURATED FATTY ACID" The following statement is a full description of this invention, including the best method of performing it known to me/us:- S019138 1 1
,/N
i 24/12/90 0 0 V 1 r 3 cc C Crt 4 4 4 AI4 r Ic 4 4 4t#cr 4 4.
I ti 4 4# 44r 444 4444 #4 4 4 44 4i 4 44 4444 la A process for the production of polyunsaturated fatty acids This invention relates to a process for the production of polyunsaturated fatty acids, such as for example arachidonic acid or eicosapentaenoic acid.
Polyunsaturated fatty acids are being used to an increasing extent in the food or cosmetic industry due in particular to their remarkable properties.
10 Arachidonic acid, for example, plays an important part as a precursor of leucotrienes and prostaglandins which are essential elements for the regulation of certain cell and physiological functions. Eicosapentaenoic acid seems to play an important part in preventing risks of thrombosis 15 or arteriosclerosis.
It is Known, for example from EP 276 982, that polyunsaturated fatty acids can be produced by enzymatic reaction using a microorganism of the Mortierella type or even by culture of the same microorganism in the presence of a hydrocarbon and a fatty acid, as described in EP 276 541.
Another process for the production of unsaturated fatty acids, which is described in EP 304 049, comprises cultivating a microorganism of the Conidiobolus type on a medium containing an unsaturated fatty acid as carbon 25 source. Yet another process, which is described in EP 277 747, comprises producing eicosapentaenoic acid from algae belonging to the Corallina type.
The object of the present invention is to provide a simple process for the production of large quantities of polyunsaturated fatty acids from natural sources, such as mosses.
To this end, the process according to the invention is characterized in that cells of a moss selected from a group i:ol- I i:r z;i P3 i i i1 :ni j ii i ti i i consisting of the species Eurhynchium striatum, Brachythecium rutabulum, Brachythecium salebrosum, Rhytidiadelphus squarrosus, Scleropodium purum and Rhytidiadelphus triquetrus are cultured and the desired polyunsaturated fatty acids are extracted therefrom.
One advantage of the present invention is that it enables a stabilized culture of cells of the selected mosses to be obtained, the said. culture producing the C" desired polyunsaturated fatty acids in a constant and reproducible manner.
Another advantage of the present invention is that it enables unsaturated fatty acids generally attached to 1 glycolipids particularly suitable for use in cosmetic 'products to be obtained.
t C *14; 9e 9 9. 4 9.
9 0 *o 9 *r 90 The mosses generally consist of two parts, namely: a haploid or gametophyte phase and a diploid or sporophyte phase. Haploid spores are produced by the sporophyte after meiosis. These spores can develop by germination to give the gametophyte.
10 To carry out the process according to the invention, the capsules containing the said spores may be collected, particularly when the spores are in the maturing phase.
The capsules collected mal be sterilized, for example by immersion in a dilute solution of sodium chloride, followed by rapid immersion in ethanol and rinsing with distilled water or by any other means capable of eliminating any germs present at their surface.
The sterilized capsules may be opened and the spores present in them may be disposed on a culture medium typically used in the culture of spores, for example a Murashige and Skoog medium (Murashige T. and Skoog F. (1962), Physipl. Plant. 15, 473-497), preferably diluted to 8-12% by weight. It is preferred to use a culture medium in semisolid form containing, for example, 1-2% by weight agar and free from growth hormones and vitamins. The spores may
I
?'s i :ii r 3 be cultivated on that medium for 1 to 4 months, preferably at a temperature of 15-30°C and in light of 2,000-7,000 lux for a period of 14-18 hours, fcllowed by a period of darkness of 6-10 hours, so that they can develop into cells.
The cells thus obtained may be recovered and inoculated in a liquid culture medium, such as for example a Murashige and Skoog medium. This medium may be used as such to allow an increase in the total lipid fraction or may be diluted to 8-12% by weight to improve the accumulation of biomass. The culture medium is preferably free g l from growth hormones and vitamins, but may contain 0 to g glucose per litre culture medium. For example, 0.1-0.8 mg cell dry matter may be inoculated per ml culture medium.
I l The pH value of the liquid culture medium may be adjusted to a value of the order of 5.5 to 7.0, for example by addition of a biological buffer solution typically used in tissue culture. This stabilization of the pH value provides for rapid differentiation of the cells and for greater production of biomass. For example, 0.05 to 0.2% by weight 2-morpholinoethanesulfonic acid may be added.
SThe cells may be left to incubate for 2 to 6 weeks at a tc temperature of 15-30'C and in light of 2,000 to 7,000 lux for about 14-18 h, followed by 6-10 h darkness. The medium c may be agitated throughout, or for only part of, the period of incubation, for example with a rotary stirrer turning at 100-120 revolutions per minute.
After incubation, the cells may be collected, for example by filtration or by centrifugation.
The cells collected may then be homogenized, for example in a solution containing 1-3 parts by weight chloroform to 1 part by weight methanol, in a hexane solution or in an isopropanol 'solution or in a mixture of these compounds. Before extraction, the cells may be |immersed for 2 to 6 minutes in a boiling isopropanol solution to inhibit the activity of the lipases. The 'si
>V.
a B C 81* 8) a .a p 04 S* .n a, ~Ja a a extract obtained may be clarified by filtration and then dried, for example in a rotary evaporator at 35-40°C under reduced pressure. The lipid fraction thus obtained contains the desired polyunsaturated fatty acids and also other fatty acids.
The various polyunsaturated fatty acids present in the lipid fraction may be separated. For example, a transesterification may be carried out using acetyl chloride to obtain the methyl estars of the desired polyunsaturated fatty acids and the methyl esters thus obtained may be extracted, for example, with hexane. The esters thus obtained may be identified, for example by gas phase chromatography, by comparison with the known characteristics of standard compounds or by mass spectrometry.
The polyunsaturated fatty acids thus obtained may be used in food and/or cosmetic products.
The present invention is illustrated in more detail in the following Examples.
Example 1 100 mg dry matter of Rhytidiadelphus squarrosus cells are inoculated in 200 ml Murashige and Skoog liquid culture medium diluted to 10% by weight, free from growth hormones and vitamins and containing 10 g glucose per litre. The pH 25 of the culture medium is adjusted to 6.5 by addition of 0.1% by weight 2-morpholinoethanesulfonic acid. The cells are left to incubate at 20°C in light of 5,000 lux for 16 hourc, followed by 8 hours in darkness, with constant stirring at 110 revolutions per minute.
After incubation for 3 weeks, the suspension is filtered and 820 mg cell dry matter are obtained.
The cells obtained are immersed for 2 minutes in a solution of isopropanol at 80-90"C and are then taken out.
The cells are then suspended in a solution containing 2 parts chloroform to 1 part methanol. The extract obtained is filtered and then dried In a rotary evaporator at 40'C under reduced pressure and purified by extraction with 0.88% by weight KCl solution. 52.5 mg lipid fraction, i.e. 6.4% by weight of the initial quantity of cell dry matter, are obtained.
The 52.5 mg of lipid fraction are transesterified with acetyl chloride and the methyl esters thus formed are extracted with hexane. The esters obtained are separated by gas phase chromatography and are identified by comparison inter alia of their mass spectrum with the mass spectra of standard compounds.
In the following Table, the fatty acid esters are 4 designated by the number of carbon atoms and the number and position of the double bonds of the basic fatty acid.
The extract of the methyl esters comprises all the fatty acids of the lipid fraction mentioned in esterified form and has the following composition in by weight: Fatty acid ester of the particular acid in *'*relation to the total q~uantity of esterified fatty acid 16:0 17.0 16:1 2.2 18:0 0.8 18:1 1.6 18:2 16.4 j18:3 18:3 16.7 2 0: 3 5.7 20:4 32.4 20:5 5.2 5.8 mg methyl arachidonate (20:4, n-6) and 0.98 mg methyl eicosapentaenoate (20:5, n-3) are obtained.
Example 2 100 mg dry matter of cells of various mosses are inoculated as in Example 1 in a Murashige and Skoog liquid culture medium which may optionally be diluted and of which the pH value may be adjusted to 6.5 by addition of 0.1% by weight 2-morpholinoethanesulfonic acid (MES acid).
The quantities of arachidonic acid (AA) and eicosapentaenoic acid (EPA) present are determined in by weight of the particular acid in relation to the total quantity of fatty acid and in mg per g dry matter of moss cells
(II).
9 a The following results are obtained: I II (mg/g) Mosses AA EPA AA EPA 20 B. rutabulum 40.1 4.7 7.4 0.9 o9 B.salebrosum 32.2 4.1 16.7 2.1 R.triquetrus 20.2 2.0 4.8 S R.squarrosus 32.4 5.2 7.1 1.2 S.purum 41.5 2.6 16.5 L9ft S* 30 E.striatum 33.6 2.7 7.0 0.6 The liquid culture medium used for each moss is: for B.rutabulum, R.triquetrus and R.squarrosus, a Murashige and Skoog medium diluted to 10% by weight and containing MES acid (pH for B.salebrosum, an undiluted Murashige and Skoog medium containing MES acid (pH for S.purum, an undiluted Murashige and Skoog medium (pH 5.8) I 1 7 for E.striatum, a Murashige and Skoog medium diluted to 10% by weight (pH 5.8).
Example 3 20 g dry matter of R. squarrosus cells are inoculated in 40 ml Murashige and Skoog liquid culture medium free from hormones and vitamins and containing 10 g glucose per litre, pH 5.8. The cells are left to incubate at 20*C in light of 5,000 lux for 16 hours, followed by 8 hours in darkness, with constant stirring at 110 revolutions per minute. After incubation for 3 weeks, the suspension is c c filtered and 30 mg cell dry matter are obtained.
CCC
c The cells obtained are immersed for 2 minutes in a solution of isopropanol at 80-90C and are then taken CC 15 out. The cells are suspended in a solution containing 2 parts chloroform to 1 part methanol. The extract is filtered, dried and purified as in Example 1 and 4.7 mg lipid fraction, i.e. 15.6% by CCC2 C' CL 20 c c u C C t r C C t C C C CC
CCC
c C C C C weight of the initial quantity of cell dry matter, are obtained. The lipid fraction is transesterified with acetyl chloride and the methyl esters are extracted with hexane. 0.40 mg methyl arachidonate, i.e. 13.2 mg/g cell dry matter, and 0.06 mg methyl eicosapentaenoate, i.e. 2.1 mg/g cell dry matter, are obtained after separation.
k.
Claims (9)
1. A process for the production of polyunsaturated fatty acids, characterized in that cells of a moss selected from a group consisting of the species Eurhynchium striatum, Brachythecium rutabulum, Brachythecium salebrosum, Rhytidi- adelphus squarrosus, Scleropodium purum and Rhytidiadelphus triquetrus are cultured and the desired polyunsaturated fatty acids are extracted therefrom.
2. A process as claimed in claim 1, characterized in that 10 the moss cells are obtained by culture of spores at 15 to 300C in light of 2,000-7,000 lux for 14-18 hours, followed by 6-10 hours in darkness, on a semisolid culture medium over a period of 1 to 4 months.
3. A process as claimed in claim 1, characterized in that the cells are cultured on a liquid culture medium at in light of 2,000-7,000 lux for 14-18 hours, followed by 6-10 hours in darkness, over a period of 2 to 6 weeks. S•
4. A process as claimed in claim 3, characterized in that the pH of the liquid culture medium is adjusted to a value 20 of 5.5-7.0.
5. A process as claimed in claim 4, characterized in that the pH is adjusted by addition of 0.05 to 0.2% by weight 2- **9 morpholinoethanesulfonic acid.
6. A process as claimed in claim 2 or 3, characterized in that the culture medium is a Murashige and Skoog medium optionally diluted to 8-12% by weight.
7. A process as claimed in claim 3, characteriied in that the liquid culture medium is free from growth hormones and vitamins and contains 0 to 25 g glucose per litre.
8. A process as claimed in claim 1, characterized in that the polyunsaturated fatty acids are extracted from the cells with chloroform, methanol, isopropanol, hexane or Smixtures of these compounds.
9. The use of the polyunsaturated fatty acids obtained by the process claimed in claim 1 in food and/or cosmetic products. I I e v k 9 A process for the production of polyunsaturated fatty acids substantially as herein described with refezxence to any one of Examples 1 to 3. DATED this 24th day of December, 1990 SOCIETE DES PRODUITS N7STLE S.A. soa *0 *0 Attorne.y: IAN ERNST Fellow In!ri.iutc of PaeP3-11 Attorneys of Australia o f S H1E -TO N W ATIER S a..a a a a S. a a. a a a a a, sea,,, a a a aa* a o a~ 7-
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CH146/90A CH680448A5 (en) | 1990-01-17 | 1990-01-17 | |
| CH146/90 | 1990-01-17 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| AU6845690A AU6845690A (en) | 1991-07-18 |
| AU622061B2 true AU622061B2 (en) | 1992-03-26 |
Family
ID=4180300
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU68456/90A Ceased AU622061B2 (en) | 1990-01-17 | 1990-12-24 | A process for the production of polyunsaturated fatty acid |
Country Status (14)
| Country | Link |
|---|---|
| EP (1) | EP0437710A1 (en) |
| JP (1) | JPH05111384A (en) |
| AU (1) | AU622061B2 (en) |
| CA (1) | CA2032648A1 (en) |
| CH (1) | CH680448A5 (en) |
| FI (1) | FI910175A (en) |
| HU (1) | HUT61334A (en) |
| IE (1) | IE904512A1 (en) |
| IL (1) | IL96584A0 (en) |
| NO (1) | NO910018L (en) |
| NZ (1) | NZ236537A (en) |
| PT (1) | PT96492A (en) |
| RU (1) | RU2038377C1 (en) |
| ZA (1) | ZA9151B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1284345B (en) * | 1961-07-14 | 1968-11-28 | Laeis Werke Ag | Process for forming lining stones designed as ring sections for pouring pans, open ovens or the like and press for performing the process |
| JP2700741B2 (en) * | 1992-03-24 | 1998-01-21 | 財団法人平岡環境科学研究所 | Cultivation method of moss using cultured species |
| ATE327340T1 (en) * | 1997-02-20 | 2006-06-15 | Dsm Ip Assets Bv | FERMENTATIVE PRODUCTION OF RECYCLABLE MATERIALS ON AN INDUSTRIAL SCALE BY USING CHEMICALLY DEFINED MEDIA |
| WO2001038541A1 (en) * | 1999-11-25 | 2001-05-31 | Basf Plant Science Gmbh | Moss genes from physcomitrella patents encoding proteins involved in the synthesis of polyunsaturated fatty acids and lipids |
| WO2010107070A1 (en) * | 2009-03-18 | 2010-09-23 | サントリーホールディングス株式会社 | NOVEL ACETYL-CoA CARBOXYLASE |
| JP5374710B2 (en) * | 2009-08-24 | 2013-12-25 | 北陸電力株式会社 | Production method of long chain polyunsaturated fatty acids |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU8300387A (en) * | 1986-12-26 | 1988-06-30 | Sagami Chemical Research Center | Process for production of eicosapentaenoic acid |
| EP0277747A2 (en) * | 1987-01-27 | 1988-08-10 | Suntory Limited | Process for production of eicosapentaenoic acid from algae |
| AU5250790A (en) * | 1989-06-22 | 1991-01-03 | Sagami Chemical Research Center | Process for production of eicosapentaenoic acid-containing phospholipid and use of said lipid |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH01243993A (en) * | 1988-03-24 | 1989-09-28 | Suntory Ltd | Production of eicosapentaenoic acid and lipid containing said acid by tissue culture of mosses |
-
1990
- 1990-01-17 CH CH146/90A patent/CH680448A5/fr not_active IP Right Cessation
- 1990-11-30 HU HU908018A patent/HUT61334A/en unknown
- 1990-11-30 EP EP90122936A patent/EP0437710A1/en not_active Withdrawn
- 1990-12-06 IL IL96584A patent/IL96584A0/en unknown
- 1990-12-14 IE IE451290A patent/IE904512A1/en unknown
- 1990-12-19 CA CA002032648A patent/CA2032648A1/en not_active Abandoned
- 1990-12-19 NZ NZ236537A patent/NZ236537A/en unknown
- 1990-12-24 AU AU68456/90A patent/AU622061B2/en not_active Ceased
-
1991
- 1991-01-03 ZA ZA9151A patent/ZA9151B/en unknown
- 1991-01-03 NO NO91910018A patent/NO910018L/en unknown
- 1991-01-14 FI FI910175A patent/FI910175A/en not_active Application Discontinuation
- 1991-01-16 RU SU914894242A patent/RU2038377C1/en active
- 1991-01-16 PT PT96492A patent/PT96492A/en not_active Application Discontinuation
- 1991-01-16 JP JP3003160A patent/JPH05111384A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU8300387A (en) * | 1986-12-26 | 1988-06-30 | Sagami Chemical Research Center | Process for production of eicosapentaenoic acid |
| EP0277747A2 (en) * | 1987-01-27 | 1988-08-10 | Suntory Limited | Process for production of eicosapentaenoic acid from algae |
| AU5250790A (en) * | 1989-06-22 | 1991-01-03 | Sagami Chemical Research Center | Process for production of eicosapentaenoic acid-containing phospholipid and use of said lipid |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH05111384A (en) | 1993-05-07 |
| NZ236537A (en) | 1991-11-26 |
| NO910018D0 (en) | 1991-01-03 |
| IL96584A0 (en) | 1991-09-16 |
| AU6845690A (en) | 1991-07-18 |
| FI910175A0 (en) | 1991-01-14 |
| EP0437710A1 (en) | 1991-07-24 |
| FI910175A (en) | 1991-07-18 |
| CA2032648A1 (en) | 1991-07-18 |
| HUT61334A (en) | 1992-12-28 |
| HU908018D0 (en) | 1991-06-28 |
| ZA9151B (en) | 1991-10-30 |
| RU2038377C1 (en) | 1995-06-27 |
| CH680448A5 (en) | 1992-08-31 |
| NO910018L (en) | 1991-07-18 |
| PT96492A (en) | 1991-10-15 |
| IE904512A1 (en) | 1991-07-17 |
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