KR20150009219A - Novel microalgae of Odontella sp. and its use - Google Patents

Abstract

The present invention relates to novel microalgae that have a high liquid content and grow rapidly. Cell growth of the novel microalgae BS-003 (KCTC 12401BP) is rapid; and the lipid content of the novel microalgae BS-003(KCTC 12401BP) is high. Therefore, a lipophilic extract or an FAME can be produced from the novel microalgae BS-003 (KCTC 12401BP); and the novel microalgae BS-003(KCTC 12401BP) can be used for biodiesel, a cosmetic composition, or a food composition. The Odontella sp. BS-003 (KCTC 12401BP) is selectively cultured in green waves of 495-570 nm.

Description

Novel microalgae Odontella sp. And its use {Novel microalgae of Odontella sp. and its use}

The present invention relates to a novel microalga Odontella sp. BS-003 and its use.

Fatty acid methyl ester (FAME) is a pollution-free fuel produced from an organism as a raw material and has a purity of 95% or more. Because FAME is produced from renewable biomass, there is no problem of depletion of energy resources and it is called biodiesel and it is also used as fuel. Carbon dioxide, which causes global warming, is recovered during the production of biomass, so the net emissions of carbon dioxide are very small. In addition, since biodiesel has a high oxygen content (more than 10% oxygen), it has a high rate of complete combustion and can reduce particulate matters such as carcinogens. Especially, it has low toxicity and high biodegradability, . Biodiesel can be used as a raw material for lubricating oil, lubricant additive, etc. as well as alternative energy for light oil, and it can be used as various pollution-free solvents and biological pesticides. To date, the production of such biodiesel has been limited to a variety of copper, And is being produced from vegetable oils and fats and has been actively studied for the production of biodiesel using microorganisms.

To date, biodiesel extraction studies have been carried out on various types of samples such as sunflower oil, Pongamia pinnata, Madhuka Indica and canola oil. However, when biodiesel is produced using land crops, grain prices may rise and food supply shortages may occur.

On the other hand, microalgae fibers are mainly cellulose and have a relatively constant diameter than plant-based cellulose fibers. Therefore, it is possible to overcome the disadvantage of changing the physical properties of the composite material due to the cellulosic size imbalance in one fiber, which is pointed out as a disadvantage of vegetable cellulose. In addition, microalgae produce organic matter from minerals by using basic production in the ocean, that is, solar energy, and their types are so wide that they can be compared with general microorganisms. In addition, microalgae It can be said that it is big. Microalgae can be divided into cell walls and various internal materials, which generally contain a lot of fiber, though they vary depending on the species. Lipids in some species are very similar to vegetable oils and are well suited to making them biofuels. The production of biodiesel can extract oil components in proportion to the content of this fat, and produce them after catalysis and transesterification. Therefore, in the production of biodiesel using microalgae, microalgae having a high lipid content and a high growth rate are required to be excavated.

Therefore, the present inventors have found new microalgae having a high lipid content from seawater and a high growth rate, establishing a condition for selectively cultivating the microalgae, and confirming the effect of FAME generation. It was confirmed that a large amount of fatty acid was present.

It is an object of the present invention to provide a novel microalga Odontella sp. BS-003 (KCTC 12401BP).

It is another object of the present invention to provide a method for preparing a lipophilic extract using the microalgae of the present invention.

Another object of the present invention is to provide a method for producing FAME using the microalgae of the present invention.

Another object of the present invention is to provide a cosmetic composition containing the lipophilic extract according to the present invention.

It is another object of the present invention to provide a food composition containing FAME according to the present invention.

In order to achieve the above object, the present invention relates to a novel microalga Odontella sp. BS-003 (KCTC 12401BP) are available.

In order to achieve the above object, the present invention also provides a method for preparing a lipophilic extract using the microalgae of the present invention.

In order to achieve the above object, the present invention provides a method for producing FAME using the microalgae of the present invention.

In order to achieve the above object, the present invention provides a cosmetic composition containing the lipophilic extract according to the present invention.

In order to achieve the above object, another object of the present invention is to provide a food composition containing FAME according to the present invention.

The novel microalgae having a high lipid content and a high growth rate according to the present invention have high cell growth and high lipid content, so that FAME or lipophilic extract can be produced therefrom.

Fig. 1 is a graph showing the results of a comparison between the novel microalga Odontella sp. BS-003 with an optical microscope and a fluorescence microscope.
Fig. 2 is a graph showing the distribution of the micro-alga Odontella sp. The 18S ribosomal RNA base sequence of BS-003 was confirmed using the NCBI blast program.
Fig. 3 is a graph showing changes in the composition of Odontella sp. A graph showing the growth curve of BS-003:
F-2-Si (-): amount of BS-003 cells cultured in F / 2 medium except Si (silicon);
F-2: Fragmented BS-003 cell mass in F / 2 medium;
F-2-N (+): amount of BS-003 cells cultured in F / 2 medium except N (nitrogen); And
The amount of bacterium BS-003 which was cultured in F / 2 medium containing 60 to 90 g / L of Na ₂ SiO ₃ .H ₂ O to contain F / 2-Si (+): Si (silicon) more than 2 times.
FIG. 4 shows that Odontella sp. ≪ RTI ID = 0.0 > BS-003.
5 is a chart of a sea water sample cultured in four light sources:
W: a sample cultured in a white LED light source;
G: a sample cultured in a green LED light source with a center wavelength of 525 nm;
B: a sample cultured in a blue LED light source with a center wavelength of 470 nm;
R: a sample cultured in a red LED light source with a center wavelength of 660 nm; And
Below: Bacterium amount of cultured BS-003.
6 is a graph showing the mass of BS-003 cultured in four light sources.
7 is a view showing neutral lipids obtained by staining BS-003 of the present invention with nile red:
× 400: 400 times magnification; And
× 200: 200 times magnification.
8 is a graph showing the lipid content of BS-003 of the present invention.
9 is a view showing the fatty acid composition of BS-003 of the present invention.

Hereinafter, the present invention will be described in detail.

The term "FAME (fatty acid methyl esters)" or "lipophilic extract " used in the present invention refers to both vegetable oil, animal fat and waste oil or waste paper. In the present invention, micro- It is said to have made. FAME can also be described as "biodiesel ".

The present invention relates to a novel microalga Odontella sp. BS-003 (KCTC 12401BP) are available.

The Odontella sp. Since BS-003 is selectively cultured at a green wavelength of 495 to 570 nm, which is a center wavelength of 525 nm, only the strain is selectively cultured in a sample containing various species, so that the strain can dominate.

The Odontella sp. BS-003 (KCTC 12401BP) may increase growth in medium with a silicon content of at least twice the content of silicon in the conventional medium (F / 2) and may contain 60 to 90 g / L Na ₂ SiO ₃ .H ₂ Lt; RTI ID = 0.0 > O. < / RTI >

In addition, the present invention relates to the use of the Odontella sp. BS-003 (KCTC 12401BP), and extracting lipids from the strain using an organic solvent.

The organic solvent is preferably a mixed solution of chloroform and methanol, more preferably a mixed solution of 2: 1 of chloroform and methanol, but is not limited thereto.

The Odontella sp. The culture of BS-003 (KCTC 12401BP) is preferably, but not limited to, culturing in a medium containing 60 to 90 g / L Na ₂ SiO ₃ .H ₂ O.

In the samples containing various strains, Odontella sp. To selectively cultivate BS-003 (KCTC 12401BP), the sample may be treated with a green wavelength of 495 to 570 nm.

In addition,

1) Odontella sp. BS-003 (KCTC 12401BP);

2) Odontella sp. Cultured in step 1) above. BS-003 (KCTC 12401BP) by adding a saponification reagent;

3) adding a methylation reagent to the reaction product of step 2) to react;

4) Extraction reagent is added to the reaction product of step 3) to extract FAME.

The reaction of step 2) is preferably carried out at 90 to 110 ° C, the reaction of step 3) is preferably carried out at 70 to 90 ° C, and the step of cooling after the reaction of step 3) But is not limited thereto.

The extraction of FAME (fatty acid methyl ether) can be performed by various extraction methods known in the art.

The Odontella sp. The culture of BS-003 (KCTC 12401BP) is preferably, but not limited to, culturing in a medium containing 60 to 90 g / L Na ₂ SiO ₃ .H ₂ O.

In the samples containing various strains, Odontella sp. To selectively cultivate BS-003 (KCTC 12401BP), the sample may be treated with a green wavelength of 495 to 570 nm.

The FAME preferably contains an unsaturated fatty acid (omega 3 fatty acid), and the unsaturated fatty acid is more preferably EPA (eicosapentanoic acid), but is not limited thereto.

In addition, the present invention relates to the use of the Odontella sp. BS-003 (KCTC 12401BP).

The cosmetic composition of the present invention can be used as a cosmetic composition in the form of a lipid, an organic solvent, a solubilizer, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, Cosmetics such as emulsifying agents, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics Or adjuvants conventionally used in the field of dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

The external form of the cosmetic composition of the present invention contains a cosmetically or dermatologically acceptable medium or base. It may be in any form suitable for topical application, for example, as a solution, a gel, a solid, a paste anhydrous product, an emulsion obtained by dispersing the oil phase in water, a suspension, a microemulsion, a microcapsule, In the form of a non-ionic follicle dispersing agent, or in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. These compositions may be prepared according to conventional methods in the art. The composition according to the invention may also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

The cosmetic composition of the present invention is not particularly limited in the form of the cosmetic composition of the present invention. For example, the cosmetic composition of the present invention can be used in a variety of forms such as a softening agent, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, Water, a pack, a powder, a body lotion, a body cream, a body oil and a body essence.

The cosmetic composition as described above may be applied to the skin, or may be applied to the skin using a micro needle or the like.

In addition, the compositions of the present invention may be made into a hair composition comprising a carrier or carrier mixture suitable for application to the hair. The carrier comprises a vehicle component commonly used in a carrier or other carrier or hair protective composition and is present in an amount ranging from about 0.5% to 99.5%, preferably from about 5.0% to 99.5%, and most preferably, About 10.0% to 90.0%. As a carrier, the solvent is selected by the copolymer to be used, regardless of whether the formulated hair composition remains on the hair after use such as hair spray, mousse, tonic, etc., or whether it is cleaned, such as shampoos, conditioners and the like. Suitable solvents for use in the present invention include water, lower alcohols (ethanol, isopropanol, etc.), hydroalcoholic mixtures, hydrocarbons such as isobutane, hexane and decene, acetone, halogenated hydrocarbons such as Freon, Dibutyl phthalate and the like), volatile silicone derivatives, siloxane (phenylpentamethyldisiloxane, methoxypropylheptamethylcyclotetrasiloxane, chloropropylpentamethyldisiloxane, hydroxypropylpentamethyldisiloxane, octamethylcyclotetrasiloxane, decamethyl Cyclopentasiloxane, and the like), and mixtures thereof.

In addition, the present invention relates to the use of Odontella sp. BS-003 (KCTC 12401BP).

The food composition preferably contains an unsaturated fatty acid, more preferably EPA (eicosapentanoic acid), but is not limited thereto.

The health functional food may be powder, granule, tablet, capsule or beverage, but is not limited thereto.

The food of the present invention can be used as it is or in combination with other food or food ingredients according to the present invention, and can be suitably used according to conventional methods.

There is no particular limitation on the kind of the food. Examples of the food to which the lipophilic extract according to the present invention can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, , Tea, a drink, an alcoholic beverage, and a vitamin complex, and includes foods in a conventional sense.

The beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

The lipophilic extract according to the present invention as an essential ingredient that can be contained in the food composition of the present invention The composition of the present invention may contain various herbal medicine extracts, food auxiliary additives, natural carbohydrates, . In addition, the food-aid additive may further include food-aid additives. Examples of the food-aid additive include food-aid additives customary in the art, for example, flavoring agents, flavoring agents, coloring agents, fillers, stabilizers and the like. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above . In addition to the above, the food composition of the present invention may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. Besides natural fruit juices and fruit juice drinks and vegetable drinks These additives may be used independently or in combination. Although the ratio of such additives is not critical, 100 parts by weight of the composition of the present invention It is generally selected from the range of 0.01 to 0.1 wt.

In a specific embodiment of the present invention, BS-003, a novel microalgae, was isolated from seawater and deposited with KCTC 12401BP. When the concentration of silicon is more than twice that of the existing concentration, the strain can be optimally cultured Respectively. In addition, in the mixed state of various papers, the microalgae of the present invention Odontella sp. It was confirmed that BS-003 can be selectively cultured by irradiating green wavelength LED. In addition, it was confirmed that the BS-003 strain of the present invention contains a large amount of neutral lipids in the cells and has a lipid content of 38% of the cell dry weight. In addition, it was confirmed that about 60% of C16 and about 20% of C18 were present in the FAME extracted from the novel strain of the present invention, and about 10% of EPA (eicosapentaenoic acid) which is an unsaturated fatty acid (omega 3 fatty acid) .

Therefore, in the novel BS-003 of the present invention having a remarkable lipid content, since the growth rate is fast at a concentration of 2 times or more of silicon, the amount of the cell is increased and the amount of the cell is increased by the selective growth at the green wavelength. From this, a lipophilic extract or FAME can be collected and used as a biodiesel, a cosmetic composition or a food composition.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Isolation and identification of microalgae from seawater

<1-1> Isolation of microalgae

The seawater samples were taken from the sea of the South Sea and then concentrated using Combi 514R (Hanil, Korea). The concentrated seawater was separated into microalgae by micropipetting and the separated microalgae were separated into 5 L bottles with F / 2 media (Table 1) at 100 rpm under a light source of 100 μmol / m ² / s Lt; / RTI &gt; Among the separated microalgae, JBS-003, which is morphologically similar to the diatom Odontella sp., Was isolated. The separated BS-003 was observed under an optical microscope (Nicon ECLIPSE 80i, nikon, Japan) and a fluorescence microscope (Nicon ECLIPSE 80i, nikon, Japan).

As a result, the morphologically Odontella aurita (Fig. 1).

<1-2> Identification of isolated microorganisms

A genetic method was used to identify the diatom BS-003 isolated in Example <1-1>.

Specifically, using the primer pairs of SEQ ID NOS: 2 and 3 shown in Table 2 below, the mixture for PCR was synthesized as shown in Table 3 below, and GeneAmp PCR system 2700 (ABI, USA) Was used to identify 18S rRNA (SEQ ID NO: 1) of BS-003 of diatom BS-003.

The 18S rRNA sequence of the identified BS-003 was confirmed by ncbi blast program, and Odontella it was identified as 96% similar to aurita to be a novel microalga (Fig. 2).

Name of sequence order SEQ ID NO: D512F ATT CCA GCT CCA ATA GCG 2 D978R GAC TAC GAT GGT ATC TAA TC 3

The composition of the reaction mixture DNA One D512F primer (12.5 pmol) One D978R primer (12.5 pmol) One 10 mM dNTP mixture One 10x PCR buffer 5 Bovine serum albumin (BSA) 0.5 Taq DNA polymerase (Takara, Japan) 0.5 dH ₂ 0 18.5 Total volume 25

Temperature() Time (minutes) Number of cycles Initial denaturation (pre-denaturation) 95 5 One Denaturation 94 0.5 30 Annealing 59 0.5 Extension 72 0.5 Post-elongation 72 7 One

Therefore, the microalgae BS-003 isolated in the present invention was classified into Odontella sp. BS-003, and deposited it on KCTC (KCTC 12401BP) on April 10, 2013.

< Example  2> BS Culture confirmation according to the medium composition of -003 strain

<2-1> Search for optimal culture medium composition

In order to establish the optimal culture condition of strain BS-003 isolated in Example 1, the growth rate of the strain was examined while adjusting the concentration of nitrogen and silicon in the composition of F / 2 medium.

Specifically, F / 2-si (-) medium in which no silicon was added to F / 2 medium, F / 2-N (+) medium (150 g / L in which nitrogen was twice the nitrogen concentration of F / Of Na ₂ SiO _3. H ₂ O) containing F / 2-si (+) medium (containing 60 to 90 g / L of NaNO ₃ ) and F / 2-si ) Was used to isolate Odontella sp. BS-003 was cultured and the degree of growth was confirmed.

As a result, growth of microalgae was slowed in the medium in which the silicon was removed, and when the content of silicon was added, the biomass productivity (amount of bacterium) was increased. The addition of an amount of nitrogen did not affect biomass production. In addition, the growth rate of the medium containing two times of silicon was 0.52, which was about 50% higher than that of the medium without silicon (Table 5 and FIG. 3).

<2-2> Determination of lipid production and biomass amount according to the culture composition of the strain

The strain BS-003 of the present invention was cultured in F / 2-si (+) medium at a temperature of 15 to 18 ° C, a light source of 100 μmol / m ² / s, stirring at 120 rpm, Biomass was identified by batch culture in a glass photobioreactor (Biotron GX, Hanil science medical, Korea). From the biomass produced, a lipid component was eluted with a 2: 1 mixture of chloroform and methanol, followed by evaporation of water in an evaporator and measurement of lipid production by measuring the weight of the lipid component.

As a result, biomass was produced at 1.83 g / L for 5 days and Odontella sp. The biomass production of BS-003 was 0.342 g / L / day (FIG. 4 and Table 5). In addition, lipid production was 0.13 g / L / day when using a batch-type incubator (Table 6).

Specific growth rate (μ) Generation time (day) F / 2-Si (-) 0.346331 2.000975 F / 2 0.475909 1.45616 F / 2-N (+) 0.468361 1.479627 F / 2-Si (+) 0.519051 1.335129

Specific growth rate (μ) Generation time (day) Biomass DCW (g / L) Lipid content (%) Biomass productivity
(g / L / day) Lipid porductivity
(g / L / day) 0.54 1.27 1.83 38 0.342 0.12996

Therefore, Odontella sp. It was found that silicon was essential for culturing BS-003.

< Example  3> BS -003 Establishment of system for selective culture of strains

It was predicted that there would be a difference in growth depending on the microalgae, and the growth of BS-003 was confirmed after irradiating specific wavelengths to selectively cultivate only the diatoms of the present invention in a mixed state of various papers. Most photosynthetic microalgae do not utilize the green wavelength most, and diatoms are known to use blue wavelength relatively more than other green algae and red algae.

Specifically, a sample containing strains BS-003 and other microalgae was irradiated with white light of 495 to 570 nm (center wavelength of about 525 nm), 450 to 495 nm of blue light of center wavelength of about 470 nm ) And a red LED having a wavelength of 620 to 780 nm (center wavelength: about 660 nm), and a 50 μmol / m ² / s light source.

As a result, the highest biomass was obtained at blue wavelength, but it was due to contamination by green algae, and green algae contamination was also observed at red and white wavelengths. However, in the green wavelength, the amount of biomass was relatively small compared to other wavelengths, but it was confirmed that only the microalga BS-003 of the present invention was cultured without contamination of other algae (FIGS. 5 and 6).

Therefore, in a sample cultured by irradiation with a green LED, Odontella sp. Since BS-003 is dominant and other algae are not cultured, it was found that BS-003 of the present invention can be selectively cultured by using LED of green wavelength.

< Example  4> BS -003 Confirmation of the amount of fat in the strain

<4-1> BS -003 Identification of neutral lipids in strains

1 μl of a solution of acetone 10 μg / L dissolved in nile red powder was added to 1 ml of culture solution of BS-003 strain and mixed for about 1 minute. After the reaction, staining of the neutrophil was confirmed by nile red staining using a Nikon ECLIPSE 80i (nikon, Japan) fluorescence microscope. All experiments proceeded in the absence of light.

As a result, Odontella sp. It was observed that a large amount of neutral lipids was contained in the cells of BS-003 (Fig. 7).

<4-2> BS -003 Confirmation of total lipid content of strains

Chlamydomonas , a green algae used for the extraction of biodiesel, reinhardtii and Chlorella vulgaris and Odontella sp. After the strain BS-003 was cultured, 10 ml of a culture solution of about 5.0 + E07 was centrifuged to collect the pellet. 15 ml of a chloroform (CH) / methanol (ME) (2: 1) solvent was added to the collected pellets and the cells were disrupted for 5 minutes using an ultrasonicator. Lipids were extracted from the disrupted cells for 15 minutes using a 300 rpm funnel shaker. To the extracted solution, 10 ml of DDW was added, and the mixture was centrifuged to separate the organic solvent and the water layer. Then, the organic solvent layer was added to the 20 ml vial. The organic solvent in the organic solvent layer was completely evaporated with an evaporator. Then, the dried weight was measured to calculate the lipid content with respect to the dry weight of the cells.

As a result, the lipid content was analyzed by weight to find that the lipid content was 38% of the cell dry weight, and the other microalgae, Chlamydomonas reinhardtii and Chlorella vulgaris And showed a lipid content of about 150 to 180% higher than that of the strains (Fig. 8).

< Example  5> BS -003 strains FAME Composition analysis

FAME was extracted from BS-003 strain of the present invention and its composition was analyzed.

Specifically, 5 ml of BS-003 strain culture was taken and centrifuged to collect microalgae. The collected cells were transferred to a glass tube and 1 ml of reagent 1 in Table 7 was added. After that, vortexing was performed so that microalgae cells were well dispersed and reacted at 100 ° C for 5 minutes. After that, vortexing was performed again and reacted for 25 minutes. When the temperature of the reaction solution dropped to about room temperature, 2 ml of reagent 2 was added, vortexed, reacted at 80 ° C for 10 minutes, and rapidly cooled. 1.25 ml of reagent 3 was added to the cooled reaction solution, and FAME was extracted by stirring for 10 minutes. Then, 3 ml of reagent 4 was added to wash the extracted FAME, and 1 ml of the washed FAME was filtered and transferred to a GC vial using a PTFE (Polytetrafluoroethylene) organic solvent filter. 50 μl of internal standard C17 (manufactured by Fluka) was added to the transferred sample to prepare a sample for FAME composition analysis. FAME analysis was performed using a gas chromatograph (Shimadzu GC-2010, Japan) and a wax column (Zebron, ZB-WAX, maximum temperature: 250 ° C, phonomenex, USA) and a flame ionization detector ) Was used to detect FAME. The injection volume used for detection was 1 μl, and the total detection time was limited to 25 minutes. SupEMCO's FAME mix 18918 (c8 to c24) was used as a standard material for biodiesel analysis. After the analysis, the peak point of each sample was compared with the peak point of the reference material, and the biodiesel value according to the reaction conditions was quantified.

Reagent 1 Saponification Reagent 45 g of NaOH, 150 ml of methanol, 150 ml of DDW Reagent 2 Methylation Reagent 325 ml of 6N HCl, 275 ml of methanol Reagent 3 Extraction Solvent Hexane 200 mL, Methyl tert-butyl ether 200 mL Reagent 4 Base Wash 10.8 g of NaOH, 900 ml of DDW

As a result, it was confirmed that 16% of FAME was extracted from the BS-003 strain. As a result of analysis of the components of the extracted FAME, C16 was present in about 60% of the total fatty acids, and C18 was about 20% in the fatty acid composition. In addition, it is confirmed that EPA (eicosapentaenoic acid), which is known to be effective for preventing various diseases such as cholesterol lowering and brain function promotion by unsaturated fatty acid (omega 3 fatty acid), which should be consumed through food with DHA and DPA, (Fig. 9).

Korea Biotechnology Research Institute KCTC12401BP 20130410

<110> Korea Research Institute of Bioscience and Biotechnology <120> Novel microalgae of Odontella sp. and its use <130> p130710 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 445 <212> DNA <213> Odontella <400> 1 tgactacgan tgtatctaat catcttcgat ccyccaactt tcgttcttga ttaatgaaaa 60 catccttggt aaatgctttc gcagtagttc atcttccgta aatccaagaa tttcacctct 120 gacaacggaa tatgaatacc cccgactgtt cctattaatc attacttcgg cgcgcaaacc 180 aacaaaatag taccggagtc ctatcttatt attccatgct aatatattca acggcataag 240 cctgctttga acactctaat tttttcacag taaacgatgg ggatcccctg cacgacaacc 300 taatgccaca caggctctcc ccaaggatgg ccggagtcaa ttgaagtacc cgcacagagt 360 gcgagaccca ttgctcccac cagatatcca actacgagct ttttaactgc aacaacttta 420 atatacgcta ttgaaactgg aataa 445 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> D512F forward primer <400> 2 attccagctc caatagcg 18 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> D978R reverse primer <400> 3 gactacgatg gtatctaatc 20

Claims (14)

A new microalgae Odontella sp. With ability to produce fatty acid methyl ester (FAME). BS-003 (KCTC 12401BP).
2. The method of claim 1, wherein the Odontella sp. BS-003 (KCTC 12401BP) is cultivated selectively at a green wavelength of 495 to 570 nm. The novel microalga Odontella sp. BS-003 (KCTC 12401BP).
2. The method of claim 1, wherein the Odontella sp. BS-003 (KCTC 12401BP) is a novel microalga Odontella sp. Strain characterized by an increase in growth medium containing 60 to 90 g / L Na ₂ SiO ₃ .H ₂ O. BS-003 (KCTC 12401BP).
The odontella sp. BS-003 (KCTC 12401BP), and extracting lipids using an organic solvent.
5. The method according to claim 4, wherein the organic solvent is a mixed solution of chloroform and methanol.
5. The method according to claim 4, wherein the Odontella sp. Wherein the culture of BS-003 (KCTC 12401BP) is cultured in a medium containing 60 to 90 g / L Na ₂ SiO ₃ .H ₂ O.
5. The method according to claim 4, wherein the Odontella sp. The culture of BS-003 (KCTC 12401BP) was carried out by irradiating the sample with a green wavelength of 495 to 570 nm to obtain Odontella sp. BS-003 (KCTC 12401BP). &Lt; / RTI &gt;
1) Odontella sp. BS-003 (KCTC 12401BP);
2) Odontella sp. Cultured in step 1) above. BS-003 (KCTC 12401BP) by adding a saponification reagent;
3) adding a methylation reagent to the reaction product of step 2) to react;
4) Extracting FAME by adding an extraction reagent to the reaction product of step 3).
The process for producing FAME according to claim 8, wherein the reaction of step 2) is carried out at 90 to 110 ° C, and the reaction of step 3) is carried out at 70 to 90 ° C.
The process for producing FAME according to claim 8, wherein the reaction is carried out after the reaction of step 3).
9. The method of claim 8, wherein the Odontella sp. Wherein the culture of BS-003 (KCTC 12401BP) is cultured in a medium containing 60 to 90 g / L Na ₂ SiO ₃ .H ₂ O.
9. The method of claim 8, wherein the Odontella sp. The culture of BS-003 (KCTC 12401BP) was carried out by irradiating the sample with a green wavelength of 495 to 570 nm to obtain Odontella sp. BS-003 (KCTC 12401BP) is selectively cultured.
A cosmetic composition comprising a lipophilic extract prepared by the method of claim 4.
9. A food composition comprising FAME prepared by the method of claim 8.

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