KR101918271B1 - Chlorella vulgaris ABC-002 strain having excellent lipid productivity containing C16 and C18 fatty acid and cell growth rate and uses thereof - Google Patents

Abstract

The present invention relates to a Chlorella vulgaris ABC-002 strain with excellent cell growth rate at a low temperature and a room temperature and productivity of lipid including fatty acid with 16 and 18 carbon atoms, and a use thereof. A novel Chlorella vulgaris ABC-002 strain, which is capable of producing fatty acid with 16 and 18 carbon atoms and ensures high growth rate by being adequate to domestic natural environment, has been isolated. The novel Chlorella ABC-002 strain of the present invention shows high growth rate at the low temperature and the room temperature, and is capable of photoautotrophically over-accumulating lipid including fatty acid with 16 and 18 carbon atoms. In addition, the novel Chlorella vulgaris ABC-002 strain of the present invention produces oleic acid (omega 9), linoleic acid (omega 6), and alpha-linolenic acid (omega 3) which are important nutritionally. Since lipid having different content in oleic acid or alpha-linolenic acid can be produced depending on the culture temperatures, the strain can be useful for producing nutritionally useful lipid and raw materials for biofuel.

Description

Chlorella vulgaris ABC-002 strain having excellent lipid productivity and cell growth rate, including C16 and C18 fatty acids, and its use {Chlorella vulgaris ABC-002 strain having excellent lipid productivity and C18 fatty acid and cell growth rate and uses thereof}

The present invention relates to a strain of Chlorella vulgaris ABC-002 having excellent lipid productivity including C ₁₆ and C ₁₈ fatty acids and cell growth rate at low temperature and room temperature, and its use.

Microalgae are photosynthetic single cell eukaryotes of size less than 50μm, one of the oldest and most diversely differentiated organisms on earth. To date, more than 44,000 species of microalgae have been found and are being studied and utilized in the production of biofuels, vaccines, high value added foods, animal and animal feed, and so on, depending on the species characteristics. Since microalgae have very different culture conditions, lipid content, lipid species, and growth rate depending on the strains, it is most important to select strains suitable for each purpose.

Chlorella is a spherical, non-migrating microalgae with a diameter of about 2 to 10 μm. It contains a variety of essential amino acids, vitamins, minerals, antioxidants and essential fatty acids that are important for nutrition. come. Unlike ω3 and ω6 fatty acids, which are produced mostly from blue fish, such as algae, fatty acids derived from microalgae are becoming more popular because they are not fishy. Recently, some chlorella species including Chlorella norovirus and Chlorella species are considered to be suitable for the production of biodiesel based on high lipid content, and the related researches have been made more actively.

One of the biggest problems with microalgae is to choose the species with the highest growth rate and lipid content to solve the economic problem. However, in case of the species that have already been discovered and registered overseas, even if they are excellent, there is a limit to utilize in domestic due to high royalties and difficult conditions, and it is also verified that outdoor cultivation is possible in domestic environment with distinct four seasons. It should be done. Therefore, in order to keep pace with the global trend in which micro-algae industry has begun to attract attention, it is necessary to secure excellent seeds in Korea and it is necessary to analyze the characteristics of each species to make healthy foods, carbon savings, feed, It is necessary to classify them as species suitable for the purpose of.

Korean Patent No. 1323957 discloses a novel strain, Chlorella vulgaris, which can grow at low temperatures, and its use, and Korean Patent No. 1241831 discloses a novel strain, Chlorella vulgaris YSW04. However, in the present invention, Likewise, there have been no reports on the use of the strain Chlorella vulgaris ABC-002 and its uses, which have excellent lipid productivity and cell growth rate, including C ₁₆ and C ₁₈ fatty acids.

The present invention has been made in view of the above-mentioned needs. In the present invention, a strain of Chlorella vulgaris ABC-002 was isolated from a lake of Incheon in Korea and isolated. The strain of Chlorella vulgaris ABC-002 is a novel strain, (C18: 1), linoleic acid (C18: 2) and alpha-linolenic acid (C18: 3) showed high lipid productivity with high content of major fatty acids in oleic acid . In particular, when the strain was cultured at 10 ° C, it was possible to produce lipids containing 30-32% (w / w) of oleic acid and 36-38% (w / w) of alpha-linolenic acid based on the total weight of fatty acids (W / w) of oleic acid and 22 to 24% (w / w) of alpha-linolenic acid was found to be able to produce lipids on the basis of the total weight of fatty acids when the strain was cultivated at 37 ° C.

Therefore, the chlorella burglary ABC-002 strain of the present invention, which is capable of producing lipids having different contents of oleic acid or alpha-linolenic acid depending on the culture temperature condition and having excellent growth even at a low temperature of 10 ° C, is useful as a raw material for biofuels and a nutritionally useful lipid Confirming that it is very useful as a strain for production, the present invention has been completed.

In order to achieve the above object, the present invention provides a strain of Chlorella vulgaris ABC-002, which has an accession number of KCTC18580P, lipid productivity including C ₁₆ and C ₁₈ fatty acids, and cell growth rate at low temperature and room temperature.

The present invention also provides a microorganism preparation for producing a lipid comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a method for producing a lipid characterized in that the strain is cultured and lipid is separated from the culture.

The present invention also relates to a method for producing a microorganism, which comprises culturing the strain;

Separating the lipid from the culture liquid;

Transesterifying the lipid to produce a fatty acid ester; And

And converting the produced fatty acid ester into biodiesel.

The present invention also provides a food containing the strain or a culture thereof as an active ingredient.

In the present invention, a novel strain of Chlorella vulgaris ABC-002 producing C ₁₆ and C ₁₈ fatty acids, which is suitable for domestic natural environments and has a rapid growth rate, was isolated. The novel chlorella ABC-002 strain of the present invention shows a high growth rate at low temperature and room temperature, and it can accumulate lipid including C ₁₆ and C ₁₈ fatty acids in a light-independent and nutritive manner even outdoors. In addition, the novel strain of Chlorella vulgaris ABC-002 of the present invention produces oleic acid (omega 9), linoleic acid (omega 6) and alpha-linolenic acid (omega 3) which are nutritionally important, and oleic acid or alpha-linolenic acid , It is very useful as a raw material for producing biofuels and as a strain for producing lipids useful for nutrition.

Fig. 1 shows the nucleotide sequence of 18S rRNA gene of the new strain isolated in the present invention.
Fig. 2 shows the phylogenetic tree prepared based on the 18S rRNA gene sequence of the new strain isolated in the present invention.
FIG. 3 shows the growth curve (A) and Multi-Color-PAM (Heiniz Walz, Germany) after the cultivation at low temperature and room temperature to examine the growth rate of the new strain Chlorella ABC- (B) shows the result of measurement of the maximum quantum yield per temperature using
FIG. 4 is a graph showing the results of analysis of C ₁₆ fatty acid, C ₁₈ fatty acid and FAME (fatty acid methyl esters) in total fatty acids after culturing the novel strain chlorella ABC-002 isolated at the present invention at 10 ° C. and 25 ° C. .

In order to accomplish the object of the present invention, the present invention provides a strain of Chlorella vulgaris ABC-002 having an accession number of KCTC18580P, excellent lipid productivity including C ₁₆ and C ₁₈ fatty acids, and cell growth rate at low temperature and room temperature.

The Chlorella vulgaris) ABC-002 strain (Accession No. KCTC18580P) is then collected in a pond in winter, Gyeonggi January 2014, the selection of single colonies formed by incubating the agarose culture plate with TAP sweetheart sympathetic via the 18S rRNA gene sequence analysis was, it was found that the cell growth rate in the low temperature and room temperature is excellent, and the lipid producing cell line, the chlorella vulgaris (chlorella The strain ABC-002 was deposited at the Korea Research Institute of Bioscience and Biotechnology (KCTC) on June 20, 2017 (Accession No .: KCTC18580P).

In addition, the Chlorella The strain ABC-002 (accession number KCTC18580P) is a major fatty acid having a carbon number of 16 and a double bond number of 0 (16: 0 fatty acid) and a carbon number of 18 and a double bond number of 1, 2 or 3 , 18: 2, 18: 3 fatty acid).

In the strain in accordance with an embodiment of the present invention, the C ₁₆ and the major fatty acids of C ₁₈ is palmitate (palmitic acid, C16: 0) , oleic acid (oleic acid, C18: 1) , linoleic acid (linoleic acid, C18: 2) and alpha-linolenic acid (C18: 3).

In the strain in accordance with an embodiment of the invention, the main fatty acid of the C ₁₈ is oleic acid (oleic acid, C18: 1) , linoleic acid (linoleic acid, C18: 2) and alpha-linolenic acid (α-linolenic Acid, C18: 3 ), Preferably oleic acid (C18: 1) and alpha-linolenic acid (C18: 3), but is not limited thereto.

The strain according to an embodiment of the present invention is characterized in that the oleic acid is 30 to 37% (w / w), linoleic acid is 7 to 12% (w / w) and alpha-linolenic acid is 20 to 38% / w). < / RTI >

In the strain according to an embodiment of the present invention, the cell growth can be excellent at 8 to 27 ° C, preferably at a low temperature of 8 to 12 ° C, at room temperature of 23 to 27 ° C, May be excellent at 10 캜 and 25 캜, but is not limited thereto.

The present invention also provides a microorganism preparation for producing a lipid comprising the strain or a culture thereof as an active ingredient. The microbial agent may include Chlorella vulgaris (Chlorella vulgaris) ABC-002 strain (Accession No. 18580P), as an active ingredient, can be effectively used for mass production of bio-oil. The mass produced bio-oil can be used to produce biodiesel through a trans-esterification process.

The present invention also provides a method for producing a lipid characterized in that the strain is cultured and lipid is separated from the culture. The strain may accumulate lipids at high concentrations at various culturing temperatures, and the culturing temperature may be preferably 8 to 27 캜, and most preferably 10 캜 and 25 캜, but is not limited thereto. Any method known in the art can be used as a method for separating lipids from the cell culture medium. Since the cell line of the present invention accumulates lipids at a high concentration in the form of a lipid body, there is an advantage that the process after extraction and extraction can be minimized.

The method according to one embodiment of the present invention comprises culturing the strain Chlorella vulgaris ABC-002 (Accession No. 18580P) of the present invention at 8 to 12 ° C to produce oleic acid in an amount of 30 to 32% (w / w) and alpha-linolenic acid can produce lipids containing 36-38% (w / w).

The method according to one embodiment of the present invention comprises culturing the strain Chlorella vulgaris ABC-002 of the present invention (accession number 18580P) at 23 to 27 ° C to produce oleic acid in an amount of 35 to 37% (w / w) and alpha-linolenic acid can produce lipids containing 22 to 24% (w / w).

In addition,

Culturing the strain;

Separating the lipid from the culture liquid;

Transesterifying the lipid to produce a fatty acid ester; And

And converting the produced fatty acid ester into biodiesel.

With the culture conditions of the strain, the culture temperature is cultured at 10 캜 or 25 캜. The culture medium for the strain may be a commonly used medium in the art, and preferably it may be a TAP medium, but is not limited thereto.

Strain from the culture medium of the present invention C ₁₆ And C ₁₈ fatty acid may be extracted by any method known in the art. The extraction solvent may be selected from the group consisting of pentane, nucleic acid, cyclohexane, toluene, methylene chloride, ethyl acetate, acetone, chloroform, methanol and mixed solvents thereof.

The invention also vulgaris (Chlorella vulgaris ) Biodiesel can be prepared by transesterifying the triglycerides produced from strain ABC-002 (accession number 18580P) to separate the fatty acid ester and glycerol. The fatty acid ester may preferably be a fatty acid methyl ester or a fatty acid ethyl ester, but is not limited thereto. The biodiesel can be produced in the form of a methyl ester or an ethyl ester by transesterifying the fatty acid contained in the biomass. The transesterification process refers to a method of transforming a large branched molecular structure of fat contained in biomass into a small, linear chain molecule required by a regular diesel engine. As the emulsifier in the transesterification process, Triton X-100 or Tween 60 may be added, but the present invention is not limited thereto. The emulsifier can be used to stabilize the interface of the bio-oil and mix well, thereby increasing the reaction yield and thus saving the biodiesel recovery cost. In addition, sodium hydroxide or potassium hydroxide may be used as a reaction catalyst in the transesterification process, but the present invention is not limited thereto.

The present invention also provides a food containing the strain or a culture thereof as an active ingredient. The Chlorella vulgaris ABC-008 strains were prepared from the fatty acids of oleic acid (C18: 1, ω9), linoleic acid (C18: 2, ω6) and alpha-linolenic acid (C18: 3, Since the lipid content is high, the food containing the strain or its culture as an active ingredient is highly nutritious. The food can be used in the form of a processed food or can be used with other food or food ingredients as it is, or can be suitably used according to conventional methods. There is no particular limitation on the kind of the processed food. Examples of the food to which the chlorella bulgurris ABC-002 strain powder can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen noodles, gums, ice cream, Beverages, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include processed foods in the conventional sense.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1. Collection and selection of microalgae

The microalgae samples were collected from a pond in Gyeonggi-do in the winter of January 2014 by one 50-mL falcon tube. The collected samples were spread in a laboratory by dropping 100 uL on a TAP agarose culture plate containing 100 ug / mL of ampicillin. Plates were then stored for 2 weeks at 120 ° C photons / m ² / s at 10 ° C in an incubator, and single colonies of the colonies formed were sorted out. The composition of the TAP culture used in the experiment is as follows; L Tris, 0.375 g / L NH ₄ Cl, 0.1 g / L MgSO ₄ .7H ₂ O, 0.05 g / L CaCl ₂ .2H ₂ O, 0.0108 g / LK ₂ HPO ₄ , 0.0054 g / L KH ₂ PO ₄ , 1 mL / L glacial acetic acid and 1 mL / L Hutner's trace elements (50 g / L Na ₂ EDTA.H ₂ O, 22 g / L ZnSO ₄ .7H ₂ O, 11.4 g / LH ₃ BO ₃ , 5.06 g / L MnCl ₂ .4H ₂ O, 1.61 g / L CoCl ₂ .6H ₂ O, 1.57 g / L CuSO ₄ .5H ₂ O, 1.10 g / L (NH ₄ ) 6 Mo ₇ O ₂₄ .7H ₂ O and 4.99 g / L FeSO ₄ .7H ₂ O).

Example  2. Taxonomic analysis

The selected microalgae were confirmed to be novel strains, and the 18S rRNA gene sequence was analyzed to identify the microalga species. The DNA of the strain was extracted using a DNeasy Plant Mini kit (Qiagen, Hilden, Germany), and the primers 18SF (5'-CCT GGT TGA TCC TGC CAG-3 ', SEQ ID NO: 2) and 18SR '-TTG ATC CTT CTG CAG GTT CA-3', SEQ ID NO: 3) was used to read the nucleotide sequence information (FIG. Based on the NCBI database based on the obtained 18S rRNA gene sequence (SEQ ID NO: 1), it was confirmed that it was a new species which has not been reported so far, and it was judged to be highly related to chlorella species. Therefore, we examined the 18S rRNA gene information of the related species and plotted the phylogenetic tree. Based on this, it was revealed that the new microalgae belong to the strain belonging to Chlorella vulgaris (Fig. 2).

Example  3. Temperature dependence Elongation  Measure

To observe the growth rate of fresh chlorella ABC-002 by temperature, cultivation was carried out at low temperature and at room temperature. TAP medium was used as the culture medium, and independent culturing was performed twice at a temperature of 10 ° C and 25 ° C in the light of 120 μmol photons / m ² / s, respectively. Initial concentrations were started at 4 × 10 ⁶ cells / mL and measured daily using Cellometer Auto X4 from Nexcelom for 12 days. It was confirmed that the cells grew up to about 3.0 × 10 ⁸ cells / mL and 3.9 × 10 ⁸ cells / mL at each temperature, and maintained the exponential phase at about 4 days at 25 ° C. and about 8 days at 10 ° C. (Fig. 3). As a result of measuring the maximum quantum yield by temperature using Multi-Color-PAM (Heinz Walz, Germany), it was confirmed that the decrease of the maximum quantum yield by temperature was not large (FIG. 3). These results indicate that the strain has a high growth rate at low temperature and room temperature and does not significantly decrease its growth at low temperature.

Example  4. Lipid extraction and component analysis

The cultured cells were harvested by centrifugation at 3500 rpm, 25 ° C for 10 minutes. After harvesting, the cells were lyophilized for accurate weighing and extraction efficiency, and 2 mL of chloroform-methanol (2: 1, vol / vol) mixture was added to 10 mg of dried cells and vigorously mixed at room temperature. For quantitative analysis, 1 mL of internal standard substance (0.5 mg heptadecanoic acid / 1 mL chloroform) was added, and 1 mL of methanol and 300 μL of sulfuric acid were further added. After vigorously stirring for about 5 minutes, the reaction was carried out at 100 ° C for 20 minutes. After cooling at room temperature, 1 ml of distilled water was added and vigorously stirred for 5 minutes. Then, the layer was separated by centrifugation at 3500 RPM and 25 ° C for 5 minutes, and the organic phase of the lower layer was extracted using a syringe, filtered and stored in a GC-vial. The extracted FAME components were analyzed by gas chromatography (HP 6890, Agilent, USA) equipped with FID (Flame ionized detector) and the columns were analyzed with HP-INNOWAX glycol column (HP 19091N-213, Agilent, USA) And the temperature was increased from 50 ° C to 250 ° C by 15 ° C per minute. As a reference, FAME standard mix (F.A.M.E.MIX C8-C24, Supelco, USA) was used. In order to increase the reliability of the reference, the GC-MS (GC-Agilent 6890 / MSD Agilent 5973) was used to revalidate the meaning of each peak. The results of the above analysis are shown in Table 1.

Linolenic acid (ω3) at 10 ℃ and oleic acid (ω3) at 25 ℃ depending on the cultivation conditions, and it was confirmed that it produced oleic acid (ω9), α-linolenic acid (ω3), linoleic acid (ω6), palmitic acid, ). In particular, it was confirmed that the yield was higher than that of the other registered chlorella strains (see Example 5 below), which means that the total lipid content accumulated in the new strain ABC-002 is high. The total fatty acid production (mg / L / day) calculated based on the high biomass yield and yield of the strain ABC-002 is shown in FIG. It is expected that the total FAME production can reach up to about 80 mg / L / day by pure culture only without specific lipid induction conditions such as nitrogen deficiency, which is industrially advantageous for fatty acid production.

Example  5. The New strain  Comparison of Growth and Lipid Productivity of ABC-002 and Other Chlorella and Related Strain

In order to compare the novel strain ABC-002 of the present invention with the related strains patented in the past, the production of cold tolerant fatty acids was carried out in Patent Information Net Kipris (http://www.kipris.or.kr/khome/main.jsp) Microalgae were searched. Related strains include Chlorella KNUA007 (Laid-open Patent No. 2015-0113282), Mike Lactinium KNUA029 (Publication No. 10-2016-0093190), Mike Lactinium KNUA032 (Laid-open Patent No. 2016-0092775) (KNUA034, KNUA032, KNUA034, KNUA034, KNUA034, KNUA034 and KNUA034), which were comparatively similar to those described in the above patent and were compared with the KNUA007, KNUA029, KNUA032 and KNUA034 specified for the yield of fatty acid, Are shown in Table 2 below.

In the results of Table 2, it was confirmed that the total fatty acid yield of the new strain ABC-002 was about 3,340 times higher than that of chlorella bulgaris KNUA007 and about 4.6 to 7.8 times higher than that of microlactinium. However, it should be noted that the culture medium used for the experiment and the incubation period were different between TAP / 12 days for the new strain ABC-002, and BG-11/20 days for KNUA007, KNUA029, KNUA032 and KNUA034.

In order to compare ABC-002 and T. chlorella strains under the same culture conditions, UTEX265, a well-known chlorella species, was purchased from UTEX Culture Collection of Algae and cultured. The culture conditions were the same as in Example 3. As shown in FIG. 5, in the case of ABC-002, ABC-002 showed about 7 times higher growth at 10 ° C and 1.8 times higher at 25 ° C on day 12. The results of comparing the total FAME production by the same method as in Example 4 are shown in FIG. 6, which shows a high production rate of about 2.81 times and 1.7 times at 10 ° C and 25 ° C, respectively. The superiority of the new strain ABC-002 .

Korea Biotechnology Research Institute KCTC18580P 20170620

<110> ADVANCED BIOMASS R & D CENTER <120> Chlorella vulgaris ABC-002 strain having excellent lipid          C16 and C18 fatty acid and cell growth          rate and uses thereof <130> PN17217 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 1795 <212> DNA <213> Chlorella vulgaris ABC-002 <400> 1 cctggttgat cctgccagta gtcatatgct tgtctcaaag attaagccat gcatgtctaa 60 gtataaactg ctttatactg tgaaactgcg aatggctcat taaatcagct atagtttatt 120 tgatggtact tactactcgg atacccgtag taaatctaga gctaatacgt gcgtaaatcc 180 cgacttctgg aagggacgta tttattagat aaaaggccga ccgggcttct gcccgactcg 240 cggtgaatca tgataacttc acgaatcgca tggccttgtg ccggcgatgt ttcattcaaa 300 tttctgccct atcaactttt gatggtagga tagaggccta ccatggtggt aacgggtgac 360 ggaggattag ggttcgattc cggagaggga gcctgagaaa cggctaccac atccaaggaa 420 ggcagcaggc gcgcaaatta cccaatcctg acacagggag gtagtgacaa taaataacaa 480 tactgggcct tttcaggtct ggtaattgga atgagtacaa tctaaacccc ttaacgagga 540 tcaattggag ggcaagtctg gtgccagcag ccgcggtaat tccagctcca atagcgtata 600 tttaagttgc tgcagttaaa aagctcgtag ttggatttcg ggtgggacct gccggtccgc 660 cgtttcggtg tgcactggca gggctcacct tgttgccggg gacgggctcc tgggcttcac 720 tgtccgggac tcggagtcgg cgctgttact ttgagtaaat tagagtgttc aaagcaggcc 780 tacgctctga atacattagc atggaataac acgataggac tctggcctat cctgttggtc 840 tgtaggaccg gagtaatgat taagagggac agtcgggggc attcgtattt cattgtcaga 900 ggtgaaattc ttggatttat gaaagacgaa ctactgcgaa agcatttgcc aaggatgttt 960 tcattaatca agaacgaaag ttgggggctc gaagacgatt agataccgtc ctagtctcaa 1020 ccataaacga tgccgactag ggatcggcgg atgtttcttc gatgactccg ccggcacctt 1080 atgagaaatc aaagtttttg ggttccgggg ggagtatggt cgcaaggctg aaacttaaag 1140 gaattgacgg aagggcacca ccaggcgtgg agcctgcggc ttaatttgac tcaacacggg 1200 aaaacttacc aggtccagac atagtgagga ttgacagatt gagagctctt tcttgattct 1260 atgggtggtg gtgcatggcc gttcttagtt ggtgggttgc cttgtcaggt tgattccggt 1320 aacgaacgag acctcagcct gctaaatagt cacggttggt tcgccagccg gcggacttct 1380 tagagggact attggcgact agccaatgga agcatgaggc aataacaggt ctgtgatgcc 1440 cttagatgtt ctgggccgca cgcgcgctac actgatgcat tcaacgagcc tagccttggc 1500 cgagaggccc gggtaatctt cgaaactgca tcgtgatggg gatagattat tgcaattatt 1560 aatcttcaac gaggaatgcc tagtaagcgc aagtcatcag cttgcgttga ttacgtccct 1620 gccctttgta cacaccgccc gtcgctccta ccgattgggt gtgctggtga agtgttcgga 1680 ttggcgacct ggggcggtct ccgctctcgg ccgccgagaa gttcattaaa ccctcccacc 1740 tagaggaagg agaagtcgta acaaggtttc cgtaggtgaa cctgcagaag gatca 1795 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 cctggttgat cctgccag 18 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ttgatccttc tgcaggttca 20

Claims (10)

KCTC18580P, Chlorella vulgaris ABC-002 strain having excellent lipid productivity including fatty acids of C ₁₆ and C ₁₈ , low temperature of 8 to 12 ° C and cell growth rate at room temperature of 23 to 27 ° C,
The main fatty acids of the C ₁₈ is oleic acid (oleic acid, C18: 1) , linoleic acid (linoleic acid, C18: 2) and alpha-linolenic acid: and (α-linolenic acid, C18 3 ),
The content of the C ₁₈ major fatty acid is 30 to 37% (w / w) of oleic acid (C18: 1), 7 to 12% (w / w) of linoleic acid (C18: 2) C18: 3) 20 to 38% (w / w). delete delete delete A microorganism preparation for producing a lipid comprising the strain of claim 1 or a culture thereof as an active ingredient. A method for producing a lipid comprising culturing the strain of claim 1 and separating lipid from the culture. The method according to claim 6, characterized in that it contains 30 to 32% (w / w) of oleic acid and 36 to 38% (w / w) of alpha-linolenic acid based on the total weight of fatty acids cultured at 8 to 12 ° C &Lt; / RTI &gt; The method according to claim 6, characterized in that it contains 35 to 37% (w / w) of oleic acid and 22 to 24% (w / w) of alpha-linolenic acid based on the total weight of fatty acids cultured at 23 to 27 ° C &Lt; / RTI &gt; Culturing the strain of claim 1 to produce a culture;
Separating the lipid from the culture liquid;
Transesterifying the lipid to produce a fatty acid ester; And
And converting the produced fatty acid ester to biodiesel. A food comprising the strain of claim 1 or a culture thereof as an active ingredient.

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