JPH03272692A - New highly unsaturated fatty acid and production of same fatty acid or lipid containing same fatty acid - Google Patents

Abstract

PURPOSE:To obtain the title fatty acid useful as a drug, etc., in high yield by culturing a fungus capable of producing arachidonic acid in a medium containing an alkene substrate and forming a highly unsaturated fatty acid or a lipid containing the fatty acid. CONSTITUTION:A fungus [e.g. Mortierella ellongata SAM0,219 (FERM P-8,703) OR (FERM P-1,239)] is cultured in a medium containing an alkene substrate or the alkene substrate is added to a culture solution in which the fungus is being cultured and further the fungus is cultured so that a highly unsaturated fatty acid (e.g. 9,17-octadecadienoic acid) or a lipid containing the fatty acid is formed to give the objective fatty acid.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention has three or more double bonds formed by fermentation, one of which is at the 1.2 or 3 position counting from the methyl group side. The present invention relates to a method for producing a novel highly unsaturated fatty acid having preferably 16 to 22 carbon atoms.

[Prior art]

Highly unsaturated fatty acids such as γ-lylunic acid, dihomo-T-lylunic acid, and arachidonic acid can be precursors of eicosanoids such as prostaglandins, leukotrienes, and thromboxanes, and are also attracting attention because of the physiological activity of the fatty acids themselves. has been done. However, modification of other carriers, such as proteins, while maintaining the structure of highly unsaturated fatty acids,
Exchange with dicarboxylic acid, etc. could not be performed.

Therefore, if a highly unsaturated fatty acid has a double bond at the end of the methyl group, it is expected that it can be used without major changes to the structure, but there is no method for producing such a new highly unsaturated fatty acid. unknown. Therefore, there is a strong desire to develop a method for producing novel highly unsaturated fatty acids with high production efficiency.

[Problem to be solved by the invention]

Therefore, the present invention aims to provide a method that can efficiently produce a novel highly unsaturated fatty acid using an inexpensive and commonly used culture medium.

[Means to solve the problem]

As a result of various studies to achieve the above objective, the present inventors have developed a microorganism capable of producing arachidonic acid.
An alkene substrate, such as a hydrocarbon having a double bond at the ω-terminus (ω1), ω2 or ω3, in the medium or the culture solution during the culture,
This is a completely novel phenomenon in which significant production of novel highly unsaturated fatty acids that retain the double bond of the alkene substrate is observed when cultured with the addition of fatty acids, fatty acid salts, fatty acid esters, or fats and oils containing these as components. I gained knowledge.

Therefore, the present invention provides for the production of alkene substrates by culturing a microorganism capable of producing arachidonic acid in a medium supplemented with an alkene substrate, or by adding an alkene substrate to the culture medium in which the microorganism is being cultured and further culturing. A method for producing a novel highly unsaturated fatty acid, which comprises producing a novel highly unsaturated fatty acid retaining a double bond or a lipid containing the novel highly unsaturated fatty acid, and collecting the novel highly unsaturated fatty acid, and arachidonic acid. A new method of culturing a microorganism capable of producing the microorganism in a medium supplemented with an alkene substrate, or further culturing the microorganism by adding an alkene substrate to the culture medium in which the microorganism is cultured, and retaining the double bond of the alkene substrate. The present invention aims to provide a novel method for producing lipids containing highly unsaturated fatty acids, which is characterized by collecting lipids containing unsaturated fatty acids.

The present inventor further provides that, in the above manufacturing method, for example, ω
When using a substrate having a double bond in 1, sesame oil and/or
or when the microorganism is cultured in the presence of an organic solvent extract of peanut oil or sesame oil, or the effective components of the extract such as sesamin, sesaminol, episesamin and/or episesaminol, 5, 8.11. It has been found that the production of 14.19-eicosapentaenoic acid is suppressed and the production of its precursor, 8,11°14,19-eicosatetraenoic acid, is increased.

Therefore, in one embodiment of the above method of the invention,
By culturing the microorganism in a medium to which sesame oil and/or peanut oil has been added, or by further culturing by adding sesame oil and/or peanut oil to the medium in which the microorganism is being cultured, 8.11°14. 19-eicosatetraenoic acid or 8.11.14° Producing a high ratio of lipids containing 19-eicosatetraenoic acid, 8.11.14
．． 19-eicosatetraenoic acid, or 8,11.14.
Either the lipid containing 19-eicosatetraenoic acid is collected; or the microorganism is grown in a medium supplemented with an extract obtained by extracting sesame oil with an organic solvent that is substantially immiscible with sesame oil. 8.11.14.19-eicosatetraenoic acid, or 8.11.14.19- - Producing a high ratio of lipids containing f-icosatetraenoic acid, 8.1
1,14.19-eicosatetraenoic acid or 8.11.
14. Collecting the lipid containing 19-eicosatetraenoic acid; or culturing said microorganism in a medium supplemented with sesamin, sesaminol, episesamin or episesaminol alone or in combination; By adding sesamin, sesaminol, episesamin, or episesaminol alone or in combination to the medium in which 8.11.14.19-eicosatetraenoic acid or 8.11. 14. Producing a high proportion of lipids containing 19-eicosatetraenoic acid, 8.1
1,14゜19-eicosatetraenoic acid, or 8,11
, 14, 19-eicosatetraenoic acid-containing lipids are collected.

The invention further provides 9,12.17-octadecatriene 112.6, 9.12.17-octadecatetraenoic acid, 8,11.14.19-eicosatetraenoic acid, and 5,8.11 A novel fatty acid selected from the group consisting of .14.19-eicosapentaenoic acid is provided.

[Specific explanation]

In the present invention, any microorganism can be used as long as it has the ability to produce arachidonic acid and, by adding an alkene substrate, has the ability to produce a novel highly unsaturated fatty acid that retains the double bond of the substrate. Examples of such microorganisms include Mortierella genus, Conidiobolus
thora) genus, Benicillium (Penici-1)
1ium), genus Cladospo
genus Rium, genus MuCor, genus Fusarium, genus Aspergillus, genus Rhodotorula.
Genus, Endo (Bntomophthora
) genus, Echinosporangium (Bchinos ora
Examples include microorganisms belonging to the genus Nium) and the genus Subrolegnia.

In the genus Mortierella, for example, Mortierella elongata (Mortierella $) IFO8570
゜Mortierella exigua (Mortierell)
a□'') IP [l 8571, Mortierella hygrophila I
FD 5941, Mortierella albina (Mort
IFO8568 can be mentioned. All of these strains can be obtained from the Fermentation Research Institute without any restrictions.

In addition, the bacterial strain Mortierella elongata SAM 0219 (Feikoken Bokuyori et al. 87
No. 03 (Feikoken Joyori et al. No. 1239) may also be used.

New highly unsaturated fatty acids obtained by culturing microorganisms capable of producing arachidonic acid in a medium supplemented with an alkene substrate include, for example, 9,17-octadecadienoic acid, 9,12.1
7-octadecatrienoic acid, 6゜9.12.17-octadecatetraenoic acid, 8,11゜14,19-eicosatetraenoic acid, 5,8.11,14゜19-eicosapentaenoic acid, etc. can be mentioned. The above novel highly unsaturated fatty acids are produced when an alkene substrate having a double bond in ω1 is used. Therefore, by changing the type of alkene substrate, completely different novel highly unsaturated fatty acids can be produced.

In order to culture the bacterial strain used in the present invention, spores, mycelia, or a preculture solution obtained by culturing the strain in advance are inoculated into a liquid medium or a solid medium and cultured. In the case of a liquid medium, any commonly used carbon sources such as glucose, fructose, xylose, sucrose, maltose, soluble starch, molasses, glycerol, mannitol, etc. can be used, but are limited to these. It's not a thing. Nitrogen sources include natural nitrogen sources such as peptone, yeast extract, malt extract, meat extract, casamino acids, and corn stable liquor, as well as organic nitrogen sources such as urea, and inorganic nitrogen sources such as sodium nitrate, ammonium nitrate, and ammonium sulfate. can be used. In addition, inorganic salts such as phosphates, magnesium sulfate, iron sulfate, copper sulfate, and vitamins can also be used as trace nutrient sources, if necessary. There are no particular limitations on the concentration of these medium components as long as they do not impair the growth of microorganisms. In practice, the carbon source is generally 0.1
~30% by weight, preferably 1-10% by weight, with 0 nitrogen source
．． The concentration is preferably 0.01 to 5% by weight, preferably 0.1 to 2% by weight.

When culturing on a solid medium, 50~
Culture is carried out for 3 to 14 days at a temperature of 5 to 40°C, preferably 20 to 30°C, using bran, rice husks, rice bran, etc. to which 100% by weight of water has been added. In this case, a nitrogen source, inorganic salts, and micronutrient sources can be added to the medium as necessary.

One method of the present invention is characterized by accumulating novel highly unsaturated fatty acids by culturing microorganisms that inherently have the ability to produce arachidonic acid in the presence of an alkene substrate. In this case, the alkene substrate is, for example, ω
Hydrocarbons having a double bond in 1 (alkenes: 1-tetradecene, 1-pentadecene, 1-hexadecene, 1-hebutadecene, 1-octadecene, 1-nonadecene, 1-
Eicosene et al.〉, fatty acid 1- with a double bond in ω1
Tetradecenoic acid, 1-pentadecenoic acid, 1-hexadenoic acid, 1-hebutadenoic acid, 1-octadecenoic acid, 1-
nonadecenoic acid, 1-eicosenoic acid, etc., fatty acid salts and esters having a double bond in ω1, or fats and oils containing a fatty acid as a constituent component having a double bond in ω1, and alcohols having a double bond in ω1 ( 13-tetradecen-1-ol, 14-pentadecen-1-ol, 1
5-hexadecen-1-ol, 16-hebutadecen-
1-ol, 17-octadecen-1-ol, 18-
ω2. Also included are similar substrates with a double bond at ω3. The total amount of alkene substrate added was 0.0 to the medium.
01 to 10% by weight, preferably 0.5 to 10% by weight. These alkene substrates may be added before or immediately after inoculation with the production microorganism, or at both times. The addition after the start of culture may be done once, or it may be added intermittently over multiple times. Alternatively, it can be added continuously. Alternatively, the culture may be carried out using an alkene substrate as the sole carbon source, in which case most of the fatty acids introduced will be novel highly unsaturated fatty acids.

In one embodiment of the present invention, sesame oil and/or peanut oil, an organic solvent extract of sesame oil, an organic solvent extract of sesame seeds, or sesamin, which is an active ingredient in the extract,
Sesaminol, Evisesamin, Episesaminol, Sesamolin, 2-(3゜4-methylenedioxyphenyl)-
6-(3-methoxy-4-hydroxyphenyl)3.7
-dioxabicyclo[3J,0]octane, 2.6-
Bis-(3-methoxy-4-hydroxyphenyl)-3
,7-thioxabicyclo[3,3,0]octane and/or 2-(3,4-methylenedioxyphenyl)-
6-(3-methoxy-4-hydroxyphenoxy)-3
, 7-thioxabicyclo[3,3,0 When a substrate having a double bond in ω1 is used, for example, by culturing the microorganism in the presence of a lignan compound such as 3,3,0 cooctane, 8,11.14. A high proportion of lipids containing 19-eicosatetraenoic acid or 8.11,14.19-eicosatetraenoic acid is produced.

The sesame oil and peanut oil in this case may be crude products or refined products. The organic solvent extract of sesame oil can be prepared using various organic solvents that are substantially immiscible with sesame oil and capable of extracting and dissolving the active ingredients.

Examples of such organic solvents include acetone, methyl ethyl ketone, diethyl ketone, methanol, and ethanol. To obtain an extract containing active ingredients, for example, after uniformly mixing sesame oil and one of the above-mentioned solvents, the mixture is left to stand at a low temperature, phase separation is performed using a conventional method such as centrifugation, and the solvent fraction is separated. Obtained by evaporating off the solvent. According to the present invention, sesamin, sesaminol,
Lignan compounds such as episesamin and episesaminol can be used alone or in combination of two or more of them.

All of these are known compounds and can be produced commercially. In addition, in order to obtain these compounds from sesame oil extract, the extract obtained as described above is subjected to column chromatography, high performance liquid chromatography,
The target compound may be isolated by treatment according to conventional methods such as recrystallization and distillation.

The amounts of additives are approximately as follows. The total amount of sesame oil, peanut oil, or both added to the medium is 0.0
01 to 10% by weight, preferably 0.5 to 10% by weight. When adding sesame oil extract, the amount added is 3 x 10-3 to 3 x 10-'% by weight based on the medium.

When sesamin, sesaminol, evisesamin, and episesaminol are added alone or in combination, the total amount added is IX10-' to 1X10-'% by weight relative to the medium. These sesame oil, peanut oil, or other ingredients may be added before or immediately after inoculating the production microorganism, or after the start of culture, or at both times. The addition after the start of culture may be done once, or it may be added intermittently over multiple times. Alternatively, it can be added continuously.

The culture temperature is 5 to 40°C, preferably 20 to 30°C,
The pH of the medium is set to 4 to 10, preferably 6 to 9, and culture with aeration and stirring, shaking culture, or static culture is performed. Culture is usually 2
Do this for ~10 days.

By culturing in this manner, lipids containing a large amount of novel highly unsaturated fatty acids are produced and accumulated within the bacterial cells. When a liquid medium is used, novel highly unsaturated fatty acids are collected from cultured bacterial cells, for example, in the following manner.

After the cultivation is completed, cultured bacterial cells are obtained from the culture solution by conventional solid-liquid separation means such as centrifugation and filtration. Wash the bacterial cells thoroughly with water,
Preferably dry. Drying can be performed by freeze drying, air drying, etc. The dried bacterial cells are preferably extracted with an organic solvent under a nitrogen stream. Ether, hexane, methanol, ethanol, chloroform, dichloromethane, petroleum ether, etc. can be used as the organic solvent, and extraction can also be carried out by alternate extraction with methanol and petroleum ether or extraction using a single layer system of chloroform-methanol-water. Good results can be obtained. By distilling off the organic solvent from the extract under reduced pressure, lipids containing a high concentration of novel highly unsaturated fatty acids can be obtained.

Furthermore, instead of the above method, extraction can be performed using wet bacterial cells. A water-compatible solvent such as methanol or ethanol, or a water-compatible mixed solvent of these and water and/or other solvents is used. Other steps are the same as above.

The lipids obtained as described above contain various novel highly unsaturated fatty acids as constituents of lipid compounds, such as fats. It is also possible to separate these directly, but
Esters with lower alcohols, such as methyl 9,17-octadecadienoate, methyl 9.12.17-octadecatrienoate, 6°9.12.17-methyl octadecatetraenoate, 8°11,14, It is preferable to separate it as methyl 19-eicosatetraenoate, methyl 5°8.11.14.19-eicosapentaenoate, etc. By forming such an ester, it can be easily separated from other lipid components, and other fatty acids produced during culture, such as palmitic acid, oleic acid, linoleic acid, etc. esterified during esterification of saturated fatty acids). For example, in order to obtain the methyl ester of a novel highly unsaturated fatty acid, the above-mentioned extracted lipid is mixed with anhydrous methanol-hydrochloric acid,
10%, 0F3-methanol 10 to 50%, etc., at room temperature for 1 to 24 hours.

In order to recover the novel highly unsaturated fatty acid methyl ester from the above-mentioned treated solution, it is preferable to extract it with an organic solvent such as hexane, ether, or ethyl acetate.

Next, this extract is dried over anhydrous sodium sulfate or the like, and the organic solvent is distilled off, preferably under reduced pressure, to obtain a mixture mainly consisting of fatty acid esters. In addition to the desired novel highly unsaturated fatty acid methyl ester, this mixture contains fatty acid methyl esters such as palmitic acid methyl ester, stearic acid methyl ester, and oleic acid methyl ester. To isolate novel highly unsaturated fatty acid methyl esters from these fatty acid methyl ester mixtures, column chromatography,
A low temperature crystallization method, a urea clathration method, a liquid-liquid current partition chromatography, etc. can be used alone or in combination.

In order to obtain a new highly unsaturated fatty acid from the various novel highly unsaturated fatty acid methyls isolated in this way, it is sufficient to hydrolyze it with an alkali and then extract it with an organic solvent such as ether or ethyl acetate.

In addition, in order to obtain a new highly unsaturated fatty acid without converting it to its methyl ester, the above-mentioned extracted lipids are subjected to alkaline decomposition (for example, with 5% sodium hydroxide at room temperature for 2 to 3 hours).
After that, the decomposed liquid can be extracted and purified by a method commonly used for extracting and purifying fatty acids.

Next, the present invention will be explained in more detail with reference to Examples.

Add 2 ml of medium (pif 6.0) containing 4% 1-hexadecene or 4% 1-octadecene and 1% yeast extract to 10 ml per serving.
- into an Erlenmeyer flask and heated to 120°C for 20
Sterilized for minutes. Mortierella albina (Morti)
erella alpina) IFO8568 was added to each medium, and cultured with shaking at 28° C. for 7 days using a reciprocating shaker (110 rl). After culturing, the bacterial cells were collected by filtration, thoroughly washed with water, dried in a centrifugal evaporator (60°C, 2 hours), and added with 2-1 methylene chloride (2-1 anhydrous methanol salt) (10%) 2 yd! Methyl esterification was achieved by treatment at 50°C for 3 hours, resulting in n-hex f:/
After adding 4 ml' of water and extracting twice and distilling off the solvent using a centrifugal evaporator (40°C, 1 hour), the obtained fatty acid methyl ester was analyzed by gas chromatography under the following analysis conditions.

Analysis conditions: Main body: GC-9A (Shimadzu Corporation) Detector: Hydrogen flame ionization detector Column: Glass column (inner diameter 3 marks) Packing agent = 5% Advance, DS on 80/10
0 mesh Chromosarb 11 (Shimadzu Corporation) Sample injection port temperature = 240°C Column temperature: 190°C Carrier gas: Nitrogen (flow rate 65-7 minutes) Even chain 1-
By adding alkenes [1-hexadecene (C+s), 1-octadecene (C+)] to the medium, ω
It was confirmed that a new highly unsaturated fatty acid having a double bond in 1 could be produced in large quantities.

Each fatty acid was isolated by fractionating the obtained fatty acid methyl ester by high performance liquid chromatography [reverse phase column (5C,@) using acetonitrile-water (85:15) as the eluent]. Table 1 shows the production amount of fatty acids and the mass spectrometry results of the products. Further, FIG. 1 shows a gas chromatography chart of fatty acids produced under the addition of 1-hexane. A similar chart was obtained when 1-octadecene was added.

Table 1 1 - Production amount per medium of each fatty acid produced when alkenes are used as substrates (■/1) Palmitic acid Stearic acid Oleic acid γ-Ryrunnic acid Dihomo-T-Ryluronic acid Arachidonic acid 13-Tetradecenoic acid 15 -Hexadecenoic acid 17-octadecenoic acid 20:4'' 8.11.14.19-eicosatetraenoic acid The NMR data of compound 20:5'' shown in the '!R1 table were as follows.

H-MMR (CD2C1 1.46 ppm (m, 2 H, CH2)t, 6
sppm (m, 2H, CHz)2, O8pp
m (m, 6H, CH2)2.30ppm'
(t, 2H, CH2) 2.82ppm (m
, 6H, CHa) 3.63 ppm (s,
3H, CH3) 4.98ppm (m, 2H
, C=C) 5.37 ppm (m, 8H, C=
C) 5.83 ppm (m, IH, C=C)
Example 2 Medium containing 4% 1-pentadecene or 4% 1-hebutadecene and 1% yeast extract (pH 6,0>2rnl!)
- into an Erlenmeyer flask and heated to 120°C for 20
Sterilized for minutes. Mortierella albina (Morti)
100 Jll of spore liquid of IFO8568 (Rella alpina) was added to each medium, and cultured with shaking at 28° C. for 7 days using a reciprocating shaker (110 rpm). After culturing, filtration, washing with water, drying, hydrolysis, methyl esterification, and extraction were performed in the same manner as in Example 1, and the obtained fatty acid methyl ester was analyzed by gas chromatography. (The analysis was conducted under the same conditions as in Example 1.) Odd-numbered chain 1-alkene (1-pentadecene (C15)
, 1-hexadecene (C17)) was found to produce a large amount of novel odd-chain highly unsaturated fatty acids having a double bond in ω1.

Furthermore, each fatty acid methyl ester was isolated by the same method as in Example 1. Table 2 shows the production amount of each fatty acid.

Further, FIG. 2 shows a gas chromatography chart of fatty acids produced under the addition of 1-pentadecene. 1
A similar chart was obtained when 1-tadecene was added to -.

Table 2 Tetradecanoic acid Pentadecanoic acid Palmitic acid Heptadecanoic acid 9-Hebutadenoic acid Oleic acid 19:0 Nonadecanoic acid Linoleic acid T-Lilunic acid 8.11.14-Nonadecatrienoic acid 5.8.11.14-Nonadecatetraenoic acid Dihomo-γ
-Ryrunic acid Arachidonic acid 14-Pentadenoic acid 16-Hebutadenoic acid 18-Nonadecenoic acid 11, 14, 18-Nonadecatrienoic acid 8, 11, 14
, 18-nonadecatetraenoic acid Example 3 Medium containing 4% 1-hexadecene and 1% yeast extract (
pH 6,0), 1-hexadecene 4%, yeast extract 1
% and sesamin 0.01% (pH 6,0)2
- into a 10- Erlenmeyer flask, 120
Sterilized at ℃ for 20 minutes. Mortierella albina (M
ortierella alapina) IFo 85
68 spore liquids (100 IJ1) were added to each medium, and cultured with shaking at 28° C. for 7 days using a resibro shaker (110 rpm). Filtration, washing with water, drying as in Example 1,
Hydrolysis, methyl esterification and extraction were performed, and the resulting fatty acid methyl ester was analyzed by gas chromatography. Table 3 shows the results.

As is clear from Table 3, by adding sesamin to the medium or to the culture solution during culture, 5, 8.11
, 14.19-eicosapentaenoic acid was suppressed, and 8,11,14.19-eicosatetraenoic acid was produced in large quantities. This result shows that the desired fatty acid It is clear that mass production of sesame oil occurs, and in addition to sesamin, which is a component of sesame oil, sesame oil and/or peanut oil, an organic solvent extract of sesame oil, an organic solvent extract of sesame seeds, or the effective In addition to sesamin, which is an ingredient, evisesamin, episesaminol, sesamolin, 2-(3゜4-methylenedioxyphenyl)-6-(3-methoxy-4-hydroxyphenyl)-3,7-thioxabicyclo[3 ,3,0
] Octane, 2,6-bis-(3-methoxy-4-hydroxyphenyl>-3,7-thioxabicyclo[3,3
,0]octane and/or 2-(3,4-methylenedioxyphenyl)-6-(3-methoxy-4-hydroxyphenoxy)-3,7-thioxabicyclo[3,3,0
]8.11.14.19 Lignan derivatives such as octane
- It is clear that it acts on the mass production of eicosatetraenoic acid.

Example 4 Culture medium containing 1% glucose, 1% yeast extract, and 3% hi-he-1-subsen! ! (pH6,0) 2mi
' into a 10+nl Erlenmeyer flask, 1
It was sterilized at 20°C for 20 minutes. Conidiobolus heterosporus
s orus) CB 5138.57, Phytium irregulare (Yuto! 亘□translation は□) CBS 494.86,
Phytophthora infestans
ora 1nfestans) IFo 4872, Entomophthora ignobilis
is) CBS 181.60, Penicillium cyaneum (Penici-"1□Col!Con) IFO533
7. C1ados
orium herbarum) IFO30314
, Mukor Ambigas (Muc Rainbow Tank Tanhoshin) 1PO
6742, Aspergillus candidus (M20 method■
Rhodotorula gratinis) IFO8816, Rhodotorula gratinis
IFO0695, Fusarinum oxituborum (Fu
sarium oxysporum) 1PO5942
, Echinosporangium transpersalis (Bch
inos oranium transversal
is) NRRL3116, Sa rore nia parasitic force (Sa rore nia 2] bidan ca) CBS
One platinum loopful of 540.67 was inoculated into a medium, and cultured with shaking at 28° C. for 7 days using a reciprocating shaker (110 rpm). Filtration, washing with water, drying, hydrolysis, methyl esterification and extraction were performed in the same manner as in Example 1, and the obtained fatty acid methyl ester was analyzed by gas chromatography.

Table 4 shows the results.

Production of a novel highly unsaturated fatty acid was observed by culturing arachidonic acid-producing bacteria in a medium supplemented with an alkene substrate.

Table 4 5.8.11°14 per culture medium of arachidonic acid producing bacteria
, 19-eicosapentaenoic acid (20:5'') shows a gas chromatography chart of fatty acids produced under heavy production. The numbers in the figure correspond to the produced fatty acid numbers in Table 1.

FIG. 2 shows a gas chromatography chart of fatty acids produced under the addition of 1-pentadecene in Example 2. The numbers in the figure correspond to the produced fatty acid numbers in Table 2.

Claims (1)

[Claims] 1. By culturing a microorganism capable of producing arachidonic acid in a medium to which an alkene substrate has been added, or by further culturing by adding an alkene substrate to the culture medium in which the microorganism is being cultured. A method for producing highly unsaturated fatty acids, which comprises producing highly unsaturated fatty acids or lipids containing highly unsaturated fatty acids, and collecting the highly unsaturated fatty acids. 2. A microorganism capable of producing arachidonic acid is cultured in a medium supplemented with an alkene substrate, or an alkene substrate is added to the culture medium in which the microorganism is cultured, and the alkene substrate is further cultured. 1. A method for producing a lipid containing a highly unsaturated fatty acid, the method comprising collecting a lipid containing a highly unsaturated fatty acid. 3. Claim 1 or 3, characterized in that the alkene substrate is a hydrocarbon, fatty acid, fatty acid salt or fatty acid ester having a double bond at the ω terminal (ω1), ω2 or ω3, or an oil or fat containing these as a constituent component. The method described in 2. 4. The novel highly unsaturated fatty acid is 9,12,17-octadecatrienoic acid, 6,9,12,17-octadecatetraenoic acid, 8,11,14,19-eicosatetraenoic acid, or 5, Process according to claim 1 or 2, characterized in that it is 8,11,14,19-eicosapentaenoic acid. 5. By culturing the microorganism in a medium to which sesame oil and/or peanut oil has been added, or by further culturing by adding sesame oil and/or peanut oil to the medium in which the microorganism is being cultured, 8,11, producing a high ratio of lipids containing 14,19-eicosatetraenoic acid or 8,11,14,19-eicosatetraenoic acid, 8,11,
14,19-eicosatetraenoic acid, or 8,11,1
The method according to claim 1 or 2, characterized in that a lipid containing 4,19-eicosatetraenoic acid is collected. 6. Cultivate the microorganism in a medium to which an extract obtained by extracting sesame oil with an organic solvent that is substantially immiscible with sesame oil, or add the extract to a medium in which the microorganism is cultured. 8,11,14,19-eicosatetraenoic acid or a lipid containing 8,11,14,19-eicosatetraenoic acid is produced at a high ratio by adding 8,11,14,19-eicosatetraenoic acid and further culturing. ,11,14,19-
The method according to claim 1 or 2, characterized in that a lipid containing eicosatetraenoic acid or 8,11,14,19-eicosatetraenoic acid is collected. 7. Cultivate the microorganism in a medium to which sesamin, sesaminol, episesamin, or episesaminol is added alone or in combination, or add sesamin, sesaminol, episesamin, or episesaminol to the medium in which the microorganism is cultured. 8,11,14,19-eicosatetraenoic acid or lipids containing 8,11,14,19-eicosatetraenoic acid are produced in a high proportion by adding them alone or in combination and further culturing. Seshime, 8
, 11,14,19-eicosatetraenoic acid, or 8,
The method according to claim 1 or 2, characterized in that a lipid containing 11,14,19-eicosatetraenoic acid is collected. 8,9,12,17-octadecatrienoic acid, 6,9,
12,17-octadecatetraenoic acid, 8,11,14
, 19-eicosatetraenoic acid, and 5,8,11,1
A fatty acid selected from the group consisting of 4,19-eicosapentaenoic acid.