JP2019099499A - Acyl steryl glucoside, method for producing acyl steryl glucoside, composition, antioxidant, hair growth or hair restoration agent, blood lipid reduction agent, anti-obesity agent, antitumor agent and agent for prevention or treatment of atherosclerosis - Google Patents

Abstract

To provide novel acyl steryl glucoside and a method for producing the same.SOLUTION: The present invention provides acyl steryl glucoside represented by formula (I) or the like, and a method for producing acyl steryl glucoside including a separation step for separating an acyl steryl glucoside fraction containing acyl steryl glucoside from a mixture of Labyrinthula, and a purification step for purifying acyl steryl glucoside from the separated acyl steryl glucoside fraction.SELECTED DRAWING: None

Description

  The present invention relates to an acylsteryl glucoside, a method for producing an acylsteryl glucoside, a composition, an antioxidant, a hair growth or hair growth agent, a hypolipidemic agent, an anti-obesity agent, an antitumor agent and atherosclerosis. It relates to a preventive or therapeutic agent.

  Labyrinthulae are eukaryotic microorganisms that inhabit the ocean, are chloroplast-free, and are dependent single-celled organisms that degrade and absorb marine organic matter. Labyrinthula is taxonomically belonging to a protist, and together with oomycetes etc. constitutes a large phylogenetic group called stramenovires. The Labyrinthulas are composed of Labyrinthidae and Trichophytonaceae in a narrow sense, and the former is classified into Labyrinthula 1 genus, the latter is classified into 11 genera such as Auranthiochtrium, Thraustochytrium, Paellithichitrium. It is done.

  Labyrinthulas are superior in the ability to produce highly unsaturated fatty acids such as DHA, saturated fatty acids such as palmitic acid, hydrocarbons such as squalene, and lipids such as sphingolipids, and they are abundant in intracellular oil spheres (oil droplets) Can be stored in Therefore, there is a deep industrial interest as a production source of medicines, functional foods and bioenergy (Patent Document 1).

  For example, Aurantiochytrium, one of the labyrinthulas, has attracted attention as a petroleum-producing alga, and has also been developed as an alternative source of n-3 highly unsaturated fatty acids industrially produced from blue fish. It is in progress.

  As a glycolipid contained in labyrinthulae, steryl glucoside (SG) in which glucose is bound to sterol has been identified. For example, four substances have been identified as steryl glucosides in A. limacinum mh0186 strain isolated from coastal seawater on Hateruma Island, Okinawa Prefecture. Specifically, GL-A (3-ObD-glucopyranosyl-stigmasta-5,7,22-triene) and GL-B (3-ObD-glucopyranosyl-4-a-methyl-stigmasta-7,22-diene) These two steryl glucosides (SGs) are structurally determined by two-dimensional NMR, and further analyzed by high performance liquid chromatography-mass spectrometer (LC-MS) analysis to obtain GL-C (3-ObD-glucopyranosyl-stigmasta-7, 22 The presence of C.-diene) and cholesteryl glucoside (CG) is shown (FIG. 1).

  Steryl glucoside (sterol glycoside) and acyl steryl glucoside (sterol glycoside ester) are contained in the oil and fat as a natural oil and fat component, and as their use, for example, hair growth and hair growth (patent document 2) , And various uses such as reduction of blood lipids (Patent Document 3) and prevention of obesity (Patent Document 4).

  Steryl glucoside and acyl steryl glucoside are useful components that are in high demand, but are rare in nature and their sources are limited. Also, it has not been known that the labyrinthulas contain acylsteryl glucoside.

Unexamined-Japanese-Patent No. 2010-200690 JP-A-7-101835 Japanese Patent Application Laid-Open No. 7-118159 Japanese Patent Application Laid-Open No. 7-107939

This invention is made in view of said subject, The objective of this invention is providing the manufacturing method of novel acyl steryl glucoside which uses labyrinthulas as a raw material.
Another object of the present invention is to provide a composition such as a food composition, a pharmaceutical composition, a cosmetic composition and the like containing an acylsteryl glucoside derived from labyrinthulae.
Furthermore, still another object of the present invention is to provide an antioxidant, a hair growth or hair growth preparation, a blood lipid lowering agent, an anti-obesity agent, an antitumor agent, and the like comprising an acylsteryl glucoside derived from labyrinthulas as an active ingredient. An object of the present invention is to provide an agent for the prophylaxis or treatment of atherosclerosis.

  MEANS TO SOLVE THE PROBLEM As a result of earnestly research in order to solve the said subject, the present inventors discovered that a labyrinthula contains glycolipid of unknown structure other than a steryl glucoside, and came to complete this invention.

Thus, according to the present invention, the above problem is solved by an acylsteryl glucoside represented by the following structural formula (I), structural formula (II) or structural formula (III).

Further, according to the present invention, there is provided a labyrinthula preparation process for preparing a labyrinthula, a separation process for separating an acylsteryl glucoside fraction containing an acylsteryl glucoside from the labyrinthula, and the separation process And a purification step of purifying the acylsteryl glucoside from the acylsteryl glucoside fraction separated in the above.
At this time, it is preferable that the acylsteryl glucoside be a steryl glucoside esterified with docosahexaenoic acid or palmitic acid.
At this time, it is preferable that the acylsteryl glucoside be a steryl glucoside esterified with docosahexaenoic acid.
At this time, it is preferable that the acylsteryl glucoside be one or more types represented by the following structural formula (I), structural formula (II) or structural formula (III).

Further, according to the present invention, the above-mentioned problems are solved by a composition containing an acylsteryl glucoside derived from a labyrinthula and selected from a food composition, a pharmaceutical composition and a cosmetic composition.
At this time, it is preferable that the acylsteryl glucoside be esterified with steryl glucoside and docosahexaenoic acid or palmitic acid.
At this time, it is preferable that the acyl steryl glucoside be a steryl glucoside esterified with docosahexaenoic acid.
At this time, it is preferable that the acylsteryl glucoside be one or more types represented by the following structural formula (I), structural formula (II) or structural formula (III).

At this time, the composition is preferably a food composition.
At this time, the composition may be a pharmaceutical composition.
At this time, the composition is preferably a cosmetic composition.

Moreover, the said subject is solved by containing the acyl steryl glucoside derived from labyrinthula as an active ingredient according to the antioxidant of this invention.
Moreover, according to the hair growth or hair growth agent of this invention, the said subject is solved by containing the acyl steryl glucoside derived from a labyrinthula as an active ingredient.
Moreover, according to the hypolipidemic agent of the present invention, the above-mentioned subject is solved by containing a labyrinthula-derived acylsteryl glucoside as an active ingredient.
Moreover, according to the anti-obesity agent of this invention, the said subject is solved by containing the acyl steryl glucoside derived from labyrinthula as an active ingredient.
Moreover, according to the antitumor agent of the present invention, the above-mentioned subject is solved by containing the labyrinthula-derived acyl steryl glucoside as an active ingredient.
Moreover, according to the preventive or therapeutic agent for atherosclerosis of the present invention, the above-mentioned subject is solved by containing the labyrinthula-derived acylsteryl glucoside as an active ingredient.

According to the present invention, novel acylsteryl glucosides can be provided.
ADVANTAGE OF THE INVENTION According to the manufacturing method of the acyl steryl glucoside of this invention, it becomes possible to manufacture the acyl steryl glucoside which is a useful substance using labyrinthulas which are algae biomass which can be mass-cultured.
In addition, it is possible to use acylsteryl glucoside derived from labyrinthula as a composition such as a food composition, a pharmaceutical composition, a cosmetic composition and the like.
Further, according to the present invention, an antioxidant, a hair growth or hair growth agent, a blood lipid lowering agent, an anti-obesity agent, an antitumor agent, and an arteriosclerosis comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient. Can be provided for the prevention or treatment of

It is a figure which shows the steryl glucoside contained in A. limacinum mh0186 strain. It is a flowchart which shows the manufacturing method of the acyl steryl glucoside of this embodiment. It is a photograph which shows the result of the thin layer chromatography (TLC) in test 1. Unidentified glycolipids are indicated by arrows. As preparations, sitosteryl glucoside and acylsteryl glucoside were used, and a lipid extracted from 2 mg of dried cells was tested in one lane. The band was detected with orcinol sulfuric acid using chloroform / methanol / water = 65/25/4 as a developing solvent. It is the result of the high performance liquid chromatography mass spectrometer (LC-MS) in test 2. It is a figure which shows the structure of the acyl steryl glucoside specified in test 2 (DHA-GL-A, Structural formula (I)). 7 shows the results of high performance liquid chromatography mass spectrometry (LC-MS) in test 2. It is a figure which shows the structure of the acyl steryl glucoside specified in test 2 (DHA-CG, structural formula (II)). It is a graph which shows the result of having analyzed the composition of the fatty acid couple | bonded with the acyl steryl glucoside (ASG) in test 3 by gas chromatography (GC). It is a graph which shows the result of having analyzed the combination of the steryl glucoside (SG) and fatty acid in an acyl steryl glucoside (ASG) in test 4 with the high performance liquid chromatography mass spectrometer (LC-MS). It is explanatory drawing which shows the flow of the binding analysis (weak alary degradation and (beta) glucosidase (EGCrP2) process) performed by the test 5. FIG. FIG. 16 is a photograph of thin layer chromatography (TLC) showing the result of binding analysis of fatty acid and sugar of ASG in test 5. FIG. A. Test 6 Fig. 5 is a graph showing the growth curve of limacinum mh0186 cells. It is a graph which shows a time-dependent change of the glucose concentration contained in the culture solution in test 6. A. The examination was conducted in Test 6. It is a graph which shows a time-dependent change of the amount of steryl glucoside (SG) contained in the cell of limacinum mh0186. A. The examination was conducted in Test 6. It is a graph which shows a time-dependent change of the amount of the acyl steryl glucoside (ASG) contained in the cell of limacinum mh0186. It is explanatory drawing which shows the ASG synthetic | combination process estimated. (A) Hypothesis 1: TG lipase transfers fatty acid to SG. (B) Hypothesis 2: Phospholipase transfers fatty acid to SG. Change of SG and ASG amount by oleic acid addition. (A) Amount of EG (ergosteryl glucoside), (b) Amount of AEG (acyl ergosteryl glucoside), (c) Total amount of AEG. Mean ± SE (n = 3), n. d means not detected. It is explanatory drawing of the structure of the novel acyl steryl glucoside (ASG) contained in the labyrinthula identified from the test 1-6. It is a figure which shows that an acyl steryl glucoside (ASG), a neutral lipid, and a phospholipid act as a reservoir of DHA accumulation in a Labyrinth. It is a photograph which shows the result of the thin layer chromatography (TLC) which measured by test 8. FIG. It is a graph which shows the result of the high performance liquid chromatography mass spectrometry (LC-MS) which measured by the test 8. FIG. It is a graph which shows the result of the oxidation measurement of DHA-SG and DHA which examined by the test 9. FIG.

Hereinafter, an embodiment (present embodiment) of the present invention will be described with reference to FIGS. 1 to 20.
The present embodiment relates to an acylsteryl glucoside, a method for producing an acylsteryl glucoside, a composition, an antioxidant, a hair growth or hair growth agent, a hypolipidemic agent, an anti-obesity agent, an antitumor agent and atherosclerosis. The present invention relates to a preventive or therapeutic agent for

The abbreviations used in the present specification are as follows.
ASG: Acyl steryl glucoside (acyl styl gluoside)
DCW: dry cell weight
DHA: docosahexaenoic acid
EG: ergosteryl glucoside
LC-MS: liquid chromatography-mass spectrometry
LPC: lysophosphatidyl choline
LPE: lysophosphatidyl ethanolamine
PC: Phosphatidyl choline
PE: Phosphatidyl ethanolamine
SE: sterol ester (sterol ester)
SG: steryl glucoside
TG: triacylglycerol (triacylglycol)
TLC: Thin layer chromatography

<Labyrinthula>
The acylsteryl glucoside of this embodiment can be obtained by culturing, separating and purifying labyrinthula which is a marine eukaryotic microorganism.
In the present embodiment, the term "labyrinthula" includes all of microorganisms, which are classified as labyrinthulas according to the zoological and botany classifications, variants thereof and variants thereof.

  Examples of the labyrinthulae include Labyrinthula, Laturninia, Althornia, Aplanochytrium, Japonochytrium, Labyrinthuloides, and Schizochytrium, It includes Thraustochytrium, Oblongicytrium, Parietichytrium, Ulkenia, and Aurantiochytrium, preferably an Oranthiochytrium. Belonging to the Properly it is like Eau lunch Oki thorium-Rimashinamu (Aurantiochytrium limacinum) is. These may be strains distributed from a storage organization or subculture strains thereof, for example, A. Limacinum mh0186 strain is exemplified.

  Labyrinthula A. Limacinum mh0186 strain (Accession No. FERM P-19755) is isolated on the coast of Hateruma Island in Okinawa Prefecture and can be grown at a wide temperature range of 10 to 35 ° C., so the acylsteryl glucoside of the present embodiment is It can be used suitably for a manufacturing method.

The labyrinthulae are widely distributed in the ocean, especially in coastal water areas and brackish water areas such as estuaries, and may be used separately from them, and any previously-isolated labyrinthula may be used. You may use the kind.
Labyrinthulas include all variants thereof. Further, among these mutant strains, those obtained by genetic methods such as recombination, transduction, transformation and the like are also contained.

  The culture of the labyrinthulae is carried out by a conventional method in a commonly used solid medium or liquid medium. At this time, as a medium to be used, for example, glucose, fructose, sucrose, starch, glycerin etc. as a carbon source, yeast extract as a nitrogen source, corn steep liquor, polypeptone, sodium glutamate, urea, ammonium acetate, ammonium sulfate, ammonium nitrate, chloride There is no particular limitation as long as it is a medium in which ammonium, sodium nitrate and the like, potassium phosphate as the inorganic salt and the other necessary components are appropriately combined, and it is generally used for cultivating labyrinthulas. Yeast extract / glucose medium (GY medium) is used. The medium is adjusted to pH 3.0 to 8.0 after preparation, and then sterilized using an autoclave or the like. Culturing may be performed by aeration and stirring culture, shaking culture, or stationary culture at 10 to 40 ° C., preferably 15 to 35 ° C., for 1 to 14 days.

Cultivation of labyrinthulas can be carried out without irradiating light, but the open pond method using sunlight directly, the sunlight collected by a light collector is sent by an optical fiber, and it is irradiated by a culture vessel to carry out photosynthesis It may be performed by a condensing method etc.
In addition, although cultivation of the labyrinthula can be performed using, for example, a fed-batch method, culture using a flask culture or a fermenter, batch culture method, semi-batch culture method (fed-batch culture method), continuous culture method It may be performed by any liquid culture method such as perfusion culture method).
The separation of labyrinthulae is carried out, for example, by centrifugation, filtration or simple sedimentation of the culture broth.

<Steryl glucoside and acyl steryl glucoside>
(Steryl glucoside)
Steryl glucoside (SG) is one of glycolipids, and has a structure in which glucose is α- or β-glucosidically linked to a sterol skeleton. It has been suggested that steryl glucoside may function as a lipid mediator in cell differentiation and stress response. In addition, steryl glucoside is known as an active ingredient such as, for example, a therapeutic agent for viral and bacterial infections, a hair growth agent, a hair growth agent, a blood lipid lowering agent, an anti-obesity agent, an anti-stress agent, and a nutrient absorption enhancer. It is done.

(Acylsteryl glucoside)
Acyl steryl glucoside (ASG) is a generic term for substances in which fatty acid is ester-bonded to steryl glucoside (SG).
Here, in the acylsteryl glucoside of the present embodiment, docosahexaenoic acid (DHA) or palmitic acid is ester-linked to steryl glucoside (SG).

The acylsteryl glucoside of the present embodiment is a compound represented by the following structural formula (I) (DHA-GL-A), structural formula (II) (DHA-CG) or structural formula (DHA esterified to steryl glucoside (SG) III) (DHA-GL-C).

  Kill human breast cancer cell line (MCF7) better than steryl glucoside (SG) as a functional feature of acyl steryl glucoside (ASG) (Wimmerova et al, Improved enzyme-mediated synthesis and supra self-assembly of naturally occurring conjugates of β-sitosterol, Steroids 117 (2017) 38-43), to reduce elevated LDL cholesterol (Ito et al, Rice branch extract containing acetylated glucoside fraction decreases elevated blood LDL cholesterol level in obese Japanese men, The Journal of Medical Investigation, 62 (2015) 80-84), Reducing the Risk of Atherosclerosis (Nhung et al, Rice Bran Extract Reduces the Risk of Atherosclerosis in Post-Menopausal Vietnamese Women., Journal of Nutritional Science and Vitaminology, 62 (2016) 295-302) and so on.

<Method for Producing Acylsteryl Glucoside>
The method for producing the acylsteryl glucoside according to the present embodiment includes a step of preparing a labyrinthula for preparing a labyrinthula, a step of separating an acylsteryl glucoside fraction containing an acylsteryl glucoside from the labyrinthula, and the step of separating And purifying the acylsteryl glucoside from the acylsteryl glucoside fraction separated in the step.
Hereinafter, each process will be described in detail with reference to FIG.

(Labyrinthula preparation process)
In the labyrinthula preparing step, labyrinthulas to be a raw material of the acyl steryl glucoside are prepared (step S1).
Although it is preferable to use the dried product of a labyrinthula cell from a viewpoint of isolation | separation and refinement | purification of an acyl steryl glucoside as a labyrinthula used as a raw material, it is not limited to this. In this embodiment, it is preferable to use a dried product of labyrinthula obtained by freeze-drying or spray-drying a live cell of labyrinthula as a raw material.

Alternatively, after culture, viable cells of rabyrintula recovered by centrifugation, filtration, sedimentation or the like can be used as a raw material. Although Labyrinthura live cells can be used as they are after harvesting from the culture tank, they are preferably washed with water or saline. In addition, it may be used in the form of a dispersion in which Labyrinthula algal cells are dispersed in a liquid such as a culture solution or water.
Furthermore, a mechanically treated product of algal cells obtained by subjecting Labyrinthula cells to ultrasonication treatment or mechanical treatment such as homogenization may be used as the raw material. In addition, a mechanical processed product dry product obtained by subjecting a mechanically processed product to a drying process may be used as a raw material.

(Separation process)
In the separation step, an acylsteryl glucoside fraction containing an acylsteryl glucoside is separated from the labyrinthula prepared in the labyrinthula preparation step (step S2).
The method of separation (fractionation) is not particularly limited as long as it can separate the acyl steryl glucoside fraction, but suitable separation means (eg, partition extraction, gel filtration, silica gel chromatography, A method obtained by fractionating a fraction having a high content of acylsteryl glucoside by reverse phase or normal phase high performance liquid chromatography (HPLC, etc.), or an extract containing ground labyrinthula and acyl steryl glucoside The extraction mixture is filtered, separated into an extract and a residue, and the extract is concentrated and fractionated by liquid chromatography.

  When the culture solution containing the cultured labyrinthula is prepared in the labyrinthula preparation step, the labyrinthula algal cells are recovered by a known method such as centrifugation or simple sedimentation of the culture solution, membrane filtration, etc. After washing, the dried algal bodies of the labyrinthula may be used in the separation step by drying using a known drying method (vacuum lyophilization, spray drying, heating vacuum drying, etc.).

  In addition, the labyrinthula prepared in the labyrinthula preparing step may be pulverized by a known pulverizing method such as an ultrasonic crusher.

The extraction solvent used in the separation step is not particularly limited as long as it can extract the acylsteryl glucoside and subsequent purification can easily be carried out, and examples thereof include methanol, ethanol, propanol, isopropanol, butanol, propylene glycol, Alcohols such as butylene glycol, methyl acetate, ethyl acetate, acetone, chloroform, toluene, pentane, hexane, cyclohexane and the like can be used alone or in combination of two or more.
In particular, it is preferable to extract the acylsteryl glucoside using a chloroform / methanol mixed solvent. At this time, although the mixing ratio of the solvents may be set arbitrarily, it is preferable to use a solvent of chloroform: methanol = 50 vol%: 50 vol% to 90 vol%: 10 vol%, and chloroform: methanol = 60 vol%: 40 vol% to 70 vol It is more preferable to use a solvent of%: 30 vol%, and it is particularly preferable to use a Folch partition, that is, a solvent of chloroform: methanol = 2: 1 (volume ratio).

(Purification process)
In the purification step, the acylsteryl glucoside is purified from the acylsteryl glucoside fraction separated in the separation step (step S3).
The method of purification is not particularly limited as long as the target acylsteryl glucoside can be purified, and examples thereof include a method of purifying acylsteryl glucoside by an appropriate separation column or recrystallization.

<Use>
Similar to other steryl glucosides (SGs) and acyl steryl glucosides (ASGs), the acylsteryl glucosides derived from labyrinthula can be used in various applications by using their functionality.

(Antioxidant)
Polyunsaturated fatty acids such as docosahexaenoic acid (DHA) have an antioxidative effect, and the acylsteryl glucoside derived from rabyrinthula in which docosahexaenoic acid (DHA) is ester-bonded to steryl glucoside (SG) is It is possible to use as an oxidizing agent.

(Hair Stabilizer, Hair Stabilizer)
Acylsteryl glucoside derived from labyrinthula can be used as a hair growth agent or a hair growth agent (Japanese Patent Application Laid-Open No. 7-101835).

(Blood lipid lowering agent)
Acylsteryl glucoside derived from labyrinthula can be used as a hypolipidemic agent used to reduce lipids such as cholesterol, neutral lipids and phospholipids in blood (JP-A-7-118159). Publication Ito et al, The Journal of Medical Investigation, 62 (2015) 80-84).

(Anti-obesity agent)
Acylsteryl glucoside derived from labyrinthula can be used as an anti-obesity agent used for treatment or prevention of obesity (Japanese Patent Application Laid-Open No. 7-107939).

(Antitumor agent)
Acylsteryl glucoside derived from labyrinthula has an effect of killing cancer cells, and can be used as an antitumor agent (anti-breast cancer agent) for suppressing tumor growth and treating cancer (Wimmerova et al, Steroids 117 (2017) 38-43).

(A preventive or therapeutic agent for atherosclerosis)
Acylsteryl glucoside derived from Labyrinthula has an action to reduce the risk of atherosclerosis, and is used as a preventive or a therapeutic agent for atherosclerosis to prevent or treat atherosclerosis. (Nhung et al, Journal of Nutritional Science and Vitaminology, 62 (2016) 295-302).

<Composition>
Antioxidant, hair growth or hair growth agent, blood lipid lowering agent, anti-obesity agent, anti-tumor agent and prevention of atherosclerosis according to the present embodiment containing the acylsteryl glucoside derived from rabilin chula as an active ingredient Alternatively, the therapeutic agent is administered as a food composition such as a health food, a pharmaceutical composition, or a cosmetic composition and administered to a living body.

(Food composition)
The acylsteryl glucoside derived from labyrinthula of this embodiment can also be used as a food.
In the food field, the food composition of the present embodiment has the effect by blending an effective amount of a labyrinthura-derived acylsteryl glucoside as a food material with various effects in the field of food. Food compositions can be provided.
Moreover, it is also possible to provide a food composition by combining the acylsteryl glucoside derived from labyrinthura with a food for specified health use, a nutritive function food, a functional display food, a food for hospital patients, a supplement and the like.
Examples of the food composition include seasonings, processed meat products, processed agricultural products, beverages (soft drinks, alcoholic drinks, alcoholic drinks, carbonated drinks, milk drinks, fruit drinks, tea, coffee, nutritional drinks, etc.), powdered drinks (powders) Juices, powdered soups, etc., concentrated beverages, confectionery (candy (nodice), cookies, biscuits, gums, gummi, chocolate, etc.), bread, cereals, etc. can be mentioned. Further, in the case of food for specified health use, nutritive function food, functional display food and the like, it may be in the form of capsules, troches, syrups, granules, powders and the like.

  Here, the food for specified health use is a food containing a health functional ingredient that affects physiological functions etc., and can be displayed as being suitable for a specific use of health with the permission of the Commissioner of the Consumer Agency. is there.

Also, the nutritive function food is a food used for supplementation of nutritional components (vitamins, minerals), and displays the function of the nutritional components. In order to sell it as a nutritionally functional food, it is necessary for the amount of nutritional component contained in the daily intake standard amount to be within the defined upper limit value and lower limit value, and not only the display of nutritional function but also alert display etc. You also need to
In addition, the functional labeling food is a food which displays scientific basis-based functionality at the responsibility of the business operator. Information on the basis of safety and functionality etc. was sent to the Secretary General of the Consumer Agency before sales.

  In the food composition according to the present embodiment, in addition to the acylsteryl glucoside derived from labyrinthula, it is possible to freely select one or two or more kinds of ingredients that can be used for the food composition. It is. For example, all additives which can usually be used in the food field can be contained, such as various seasonings, preservatives, emulsifiers, stabilizers, flavors, coloring agents, preservatives, pH adjusters and the like.

  In the food composition according to the present embodiment, it is possible to add one or more kinds of other substances which are known to have various effects, to the acylsteryl glucoside derived from labyrinthula.

(Pharmaceutical composition)
The acylsteryl glucoside derived from labyrinthula of this embodiment can be used as a pharmaceutical composition.
The pharmaceutical composition of the present embodiment is a pharmaceutical composition having the above-mentioned action by blending a pharmaceutically acceptable carrier or additive together with an amount of the labyrinthura-derived acylsteryl glucoside capable of exerting various actions effectively. The goods are provided. The pharmaceutical composition may be a pharmaceutical or quasi-drug.
The pharmaceutical composition may be applied externally or internally. Therefore, the pharmaceutical composition is used in the form of an internal preparation, an injection such as subcutaneous injection, intravenous injection, intradermal injection, intramuscular injection and / or intraperitoneal injection, transmucosal application, transdermal application and the like. be able to.
The dosage form of the pharmaceutical composition can be appropriately selected depending on the form of application, for example, liquid preparation such as solution, suspension preparation, semi-solid preparation such as ointment or gel preparation, tablet, granules, Examples include solid preparations such as capsules, powders, powders and the like.

In the pharmaceutical composition according to the present embodiment, one or more pharmaceutically acceptable additives can be freely selected and contained.
For example, when the pharmaceutical composition according to the present embodiment is applied to an oral preparation, for example, excipients, binders, disintegrants, surfactants, preservatives, coloring agents, flavoring agents, fragrances, stabilizers, preservatives All additives which can usually be used in the field of pharmaceutical preparations such as agents, antioxidants and the like can be contained. In addition, a drug delivery system (DDS) can be used to make a sustained release preparation or the like.

  In addition to the acylsteryl glucoside derived from rabilin chula, it is also possible to add one or more kinds of other substances known to have various actions to the pharmaceutical composition according to the present embodiment.

(Cosmetic composition)
The acylsteryl glucoside derived from labyrinthula of this embodiment can be suitably used in a cosmetic composition by utilizing various effects.
The cosmetic composition can be applied to any form of cosmetic. For example, it can be applied to skin care cosmetics such as lotions, milky lotions, creams, and beauty essences, foundations, concealers, makeup bases, lipsticks, blushers, eye shadows, makeup products such as eye shadows, eyeliners, sunscreen cosmetics, etc. .

In the cosmetic composition according to the present embodiment, in addition to the acylsteryl glucoside derived from labyrinthula, one or two or more freely selected components to be used in the cosmetic composition may be selected and blended. Is possible.
For example, substrates, preservatives, emulsifiers, colorants, preservatives, surfactants, UV absorbers, antioxidants, moisturizers, UV absorbers, fragrances, preservatives and fungicides, extender pigments, color pigments, alcohols, All additives which can usually be used in the cosmetic field, such as water, can be included.

  In the cosmetic composition according to the present embodiment, the content of the acylsteryl glucoside derived from labyrinthula is not particularly limited, and can be freely set according to the purpose.

Hereinafter, the present invention will be specifically described based on specific examples, but the present invention is not limited to these.
In the following example, an unknown glycolipid is isolated from the total lipid fraction of the labyrinthula Aurantiochytrium limacinum mh0186 strain using a Sep-pak plus silica column, and the structure is determined by mass spectrometry, weak alkaline decomposition, or β-glucosidase treatment. It analyzed. Moreover, glycolipid was quantified by the liquid chromatograph mass spectrometer (LC-MS).

<Material>
SG and ASG were purchased from Matreya LCC. TG (12: 0/12: 0/12: 0), PC (11: 0/1: 1: 0), LPC (13: 0), PE (12: 0/12: 0), LPE (13: 0) , SE (16: 0-d7 cholesterol), d7 cholesterol were purchased from Avanti Polar Lipids. Artificial sea water (SEALIFE) was purchased from Japanese sea water. The organic solvent used for LC-MS analysis was purchased from Wako Pure Chemical Industries.

Test 1 Isolation and Purification of Unknown Glycolipids
(Culturing of A. limacinum mh0186)
A. limacinum mh0186 strain was used as a labyrinthula. Cells precultured with 3 ml of 0.1% vitamin mixture, 0.2% element solution in GY medium (3.1% (w / v) D-glucose, 1.0% (w / v) Yeast Extract, 1.75% (w / v) SEALIFE) 100 μl of the above was added to 100 ml of GY medium (containing 300 ml Erlenmeyer, 0.1% vitamin mixture, 0.2% element solution) and cultured with shaking for 3 days (150 rpm, 25 ° C.). Here, the composition of vitamin Mixture is, 0.2% (w / v) vitamin B 1, 0.001% (w / v) vitamin B 2, is 0.001% (w / v) vitamin B 12. The composition of the element solution is 3% (w / v) EDTA do-sodium, 0.145% (w / v) FeCl ₃ · 6H ₂ O, 3.42% (w / v) H ₃ BO ₃ , 0.43% (w / v) ) MnCl ₂ · 4 H ₂ O, 0.1355% (w / v) ZnSO ₄ · 7 H ₂ O, 0.013% (w / v) CoCl ₂ · 6 H ₂ O, 0.01% (w / v) CuSO ₄ · 5 H ₂ O is there.

(Lipid extraction)
The supernatant was removed by centrifugation (4.degree. C., 3,000.times.g, 5 min) for 100 ml of the culture broth cultured for 3 days. After washing with 1.75% SEA LIFE, the cells were recovered by centrifugation again to obtain dried cells by lyophilization. The weight of the resulting dried cells was measured, and an equal amount of water and 200 volumes of chloroform (C) / methanol (M) = 2/1 were added to extract lipids by ultrasonic wave. The solution was concentrated to dryness, added with C / M / water (W) = 8/4/3, centrifuged (600 × g, 5 min), and the lower layer was recovered and dried to obtain total lipid.

(Isolation and purification of unknown glycolipids)
Conditioning was carried out by sequentially flowing methanol, acetone and chloroform over Sep-Pak plus silica cartridge (manufactured by Waters). The collected total lipids were dissolved in 3 ml of chloroform and the whole was applied to a Sep-Pak plus silica column. After eluting the neutral lipid with 10 ml of chloroform, it was eluted with C / acetone (A) = 80: 20 (vol%).

(Thin layer chromatography (TLC) analysis)
The eluate separated on a Sep-Pak plus silica column was dried and then dissolved in C / M = 2/1, and a lipid of 2 mg DCW was loaded on a TLC plate (TLC Silica Gel 60, Merck).
C / M / W = 65/25/4 (v / v / v) was used as a developing solvent, and color was developed with orcinol sulfuric acid (0.2% orcinol, 11.4% sulfuric acid).

(Result of test 1)
When the total lipid of A. limacinum mh0186 was detected by TLC, the band of the unknown glycolipid was very close to the band of the neutral lipid, and the band became invisible because there were too many neutral lipids. Therefore, neutral lipids, glycolipids and phospholipids were separated from the total lipids of A. limacinum mh0186 using a Sep-Pak plus silica column. As a result, it was detected in the acetone fraction together with SG. Next, as a result of examining the isolation method of the unknown glycolipid which does not contain SG using a Sep-Pak plus silica column, when chloroform / acetone = 80/20 eluted, the unknown glycolipid could be isolated (FIG. 3). Also, this band was in agreement with the standard ASG Rf value.

<Test 2 LC-MS analysis of unknown glycolipids>
(Purchasing of unknown glycolipid from silica)
An unknown glycolipid was applied to a TLC plate (2 mg DCW min × 7 lanes), developed under the same conditions as in Test 1, and a band was detected. The detected band was scraped off with a spatula.

(Removal of silica using spin column)
C / M / W = 65/25/4 (v / v / v) was added to the scraped silica, and the lipid was extracted by sonication. After centrifugation (600 × g, 5 min), the supernatant was collected and dried. The suspension was added with 400 μl of C / M = 1/2 and 800 μl of M / 0.45% NaCl = 1/1 and suspended, and tested in MonoSpin C18 FF (manufactured by GL Sciences). The waste liquid was removed by centrifugation (1,000 × g, 1 min), and this operation was performed three times. The column was added with 200 μl of ultrapure water and centrifuged (1,000 × g, 1 min) to remove waste liquid. The column was added with 100 μl of methanol and centrifuged (1,000 × g, 1 min) to obtain an eluate.

(LC-MS analysis)
The unknown glycolipid was analyzed by LC-MS (3200 Q TRAP, manufactured by AB SCIEX). The mixture was poured at a flow rate of 200 μl / min, and phospholipids, neutral lipids and glycolipids were separated using InertSustain C18 (2.1 × 150 mm, 5 μl, manufactured by GL Science) on the column. Eluates are A (Acetonitrile / Methanol / H ₂ O = 19/19/2 (v / v / v), 0.1% Formic acid, 0.028% Ammonia) and buffer B (IPA, 0.1% Formic acid, 0.028% Ammonia) It was used. The conditions for the cladid are 3% B for 3 minutes, then 40% B for 21 minutes, 70% B for 1 minute, 70% B for 7 minutes, and finally 3% B for 7 minutes. did. A: 100%, B: 0% (3 min) → A: 60%, B: 40% (21 min) → A: 30%, B: 70% (1 min) → A: 30%, B: 70% (7 min) Set to).

(Result of trial 2)
The results are shown in FIGS. 4A and 5A. Both unknown glycolipids were detected in a value consistent with the mass number of the sterol backbone moiety. Further, as a result of MS / MS / MS analysis, a value corresponding to the mass number of a portion characteristic to each sterol skeleton was detected. From these results, in A. limacinum mh0186, acylsteryl glucoside (ASG) in which DHA was added to GL-A (3-ObD-glucocopyranosyl-stigmasta-5, 7, 22-triene) or CG (cholesteryl glucoside) It was revealed to exist (Fig. 4B, Fig. 5B). Specifically, DHA-GL-A (FIG. 4B, structural formula (I)) in which DHA is added to GL-A and DHA-CG (FIG. 5B, structural formula (II)) in which DHA is added to CG are It was found to exist.

Test 3 Gas Chromatography (GC) Analysis of ASG Fatty Acids
The composition of the fatty acid bound to the acyl steryl glucoside (ASG) was analyzed by gas chromatography (GC).

(Result of trial 3)
The results are shown in FIG. It was found that DHA (C22: 6) and palmitic acid (C16: 0) are abundantly present in unknown glycolipids.

<Test 4 Molecular species analysis of ASG>
The combination of steryl glucoside (SG) and fatty acid in acyl steryl glucoside (ASG) was analyzed by LC-MS.

(Result of Test 4)
The results are shown in FIG. The most frequent ASG to which DHA was added was particularly abundant DHA-CG (FIG. 5B) and DHA-GL-A (FIG. 4B). DHA-GL-C (Structural formula (III)) was also present. In addition, many SG existed in the order of GL-A, GL-B, GL-C, and CG.
Acylsteryl glucoside (ASG) to which DHA was added had a novel structure which has not been reported so far.

Test 5 Binding Analysis of ASG Fatty Acid and Sugar
From the analysis results of LC-MS, the unknown glycolipid was considered to be an acylsteryl glucoside (ASG) in which a fatty acid is bound to SG. In plants, ASG has been reported to have a structure in which a fatty acid is ester-bonded to carbon at position 6 of glucose. Therefore, ASG of A. limacinum mh0186 was similarly treated with weak alkaline decomposition and β-glucosidase (EGCrP2) to determine whether it had a structure in which fatty acid was ester-bonded to SG (FIG. 8).

(Weak alkaline decomposition)
90 μl of C / M = 1/2 and 10 μl of NaOH were added to the purified unknown glycolipid (4 mg freeze-dried cell weight) and incubated at 37 ° C. After 2 hours, the temperature was returned to normal temperature, acetic acid diluted 10-fold with 6 μl of methanol was added to neutralize, and then the solution was nitrogen-dried.

(Purification of unknown glycolipids)
The reaction unknown dried glycolipid was dissolved in 200 μl of C / M = 1/2 and desalted in the same manner as in Test 2. The extract was dried in nitrogen.

(Beta glucosidase treatment)
To a weakly alkaline-degraded unknown glycolipid, 50 mM of Acetate buffer (pH 5.0), 0.025% Sodium Chocolate and 500 ng of recombinant EGCrP2 were added to a total amount of 20 μl, and allowed to react at 37 ° C. for 24 hours. As a negative control, one to which boiled EGCrP2 was added was also prepared and reacted under the same conditions. After completion of the reaction, it was evaporated to dryness.

(TLC analysis)
The dried lipid was dissolved in 5 μl C / M = 2/1 and the whole was applied to a TLC plate. C / M / W = 65/25/4 was developed as a developing solvent and developed with orcinol sulfuric acid.

(Result of Test 5)
As a result of weak alkaline decomposition and β-glucosidase treatment, the unknown glycolipid was resistant to EGCrP2, but was degraded by EGCrP2 by weak alkaline decomposition (FIG. 9). From the results of this test, it was found that the unknown glycolipid was acylsteryl glucoside (ASG) in which fatty acid was ester-bonded to steryl glucoside (SG).

<Test 6 Comparison of SG and ASG amounts at culture time>
In Test 6, changes in the amounts of SG and ASG at the culture time of A. limacinum mh0186 were analyzed by LC-MS.

(Preparation of culture and growth curve of A. limacinum mh0186)
Cells precultured with 3 ml of GY medium (0.1% vitamin mixture, 0.2% element solution) are 100 ml of GY medium (300 ml Erlenmeyer flask, 0.1% vitamin mixture, 0.2% so that the initial concentration is OD ₆₀₀ = 0.02). In addition to the element solution), shake culture was carried out (150 rpm, 25 ° C.). The OD ₆₀₀ was measured every 24 hours.

(Measurement of glucose concentration)
One ml of culture fluid was collected every 24 hours, and centrifuged (4 ° C., 3,000 × g, 5 min) to recover the supernatant. Using ultrapure water, culture supernatant on day 1 by 100 times, supernatant on day 2 and day 3 by 50, supernatant on day 4 by 10, and supernatant on day 5 through 5 Diluted twice. In addition, a 2-fold dilution series of glucose standard solution (200 mg / dL) was prepared for preparation of a calibration curve, and aliquoted in 10 μl portions.
To each well of the 96-well plate, 10 μl of the diluted culture solution and a coloring reagent (glucose CII test, Wako) were added, and after incubating for 15 minutes at room temperature, OD ₄₉₂ was measured with MULTISKAN FC (manufactured by Thermo SCIENTIFIC).

(Cell recovery and lipid extraction)
One ml of culture medium was collected every 24 hours and centrifuged (4 ° C., 3,000 × g, 5 min) to remove the supernatant. Lyophilized cells were obtained after washing with 1.75% SEALIFE. Measure 4 mg of dried cells, 300 μl of internal standard (20 μM EG, 10 μM ASG (18: 0-sitosteryl glucoside), TG (12: 0: 12: 0/12: 0), PC (11: 0: 11: 0), LPC (13: 0), PE (12: 0/12: 0), LPE (13: 0), SE (16: 0-d7 cholesterol), d7 cholesterol) containing C / M = 2 / 1 was added and the lipids were extracted with ultrasound for 10 minutes. After centrifuging (4 ° C., 15,000 rpm, 5 min) and collecting the supernatant, 75 μl of ultrapure water was added and centrifuged (4 ° C., 15,000 rpm, 5 min) to recover the lower layer.

(LC-MS analysis)
100 μl of the recovered lower layer was added to an autosampler vial containing 900 μl of 2-propanol, and 5 μl was subjected to LC-MS. The LC-MS conditions are the same as in Test 2.

(Result of Test 6)
The results are shown in FIGS. FIG. 10 shows the growth curve of cells, FIG. 11 shows the time course of glucose concentration in the culture solution, FIG. 12 shows the time course of steryl glucoside (SG) amount, and FIG. 13 shows the amount of acylsteric glucoside (ASG) It shows a change over time.

Acyl steryl glucoside (ASG) increased in the logarithmic growth phase and decreased in the stationary phase and later, but the steryl glucoside (SG) tended to increase after the stationary phase.
From these results, DHA binds to SG and is accumulated in the body when there is a sufficient carbon source, but when glucose is depleted and nutrition is lost, DHA bound to SG is released, and energy source It was thought that it would be. Therefore, it was suggested that the acyl steryl glucoside (ASG) may act as a reservoir of steryl glucoside (SG).

<Test 7 Search for ASG synthetase in budding yeast>
With regard to ASG synthetase, two hypotheses are that TG lipase transfers fatty acid to SG (Hypothesis 1, FIG. 14 (a)), or phospholipase transfers fatty acid to SG (Hypothesis 2, FIG. 14 (b)) Thought. Of the two hypotheses, hypothesis 1 was tested using budding yeast.

(Comparison of SG and ASG amounts by oleic acid addition in budding yeast)
Budding yeast produces large oil droplets by adding oleic acid to the medium. Therefore, oleic acid was added to the medium to make large oil droplets to increase the amount of TG possessed by budding yeast, and changes in the amounts of SG and ASG were analyzed by LC-MS. In addition, since Budding yeast wild strain has almost no SG, in this test, Ugt51 which transfers Glc from UDP-Glc to ergosterol, which is the main sterol of budding yeast, and synthesizes ergosteryl glucoside (EG) An overexpressed EG overaccumulating strain was used.
(Culture of budding yeast UGT51OE / egh1KO strain)
Cells precultured with 3 ml of YPD medium (2.0% (w / v) D-glucose, 1.0% (w / v) Polypeptone, 1.0% (w / v) Yeast Extract) using UGT 51 OE / egh 1 KO strain, It was added to 3 ml of YPD medium to a concentration of OD ₆₀₀ = 0.2. To YPD medium, 3 μl of 1 μg / μl oleic acid dissolved in ethanol was added, and shaking culture was performed for 24 hours (150 rpm, 30 ° C.). As a negative control, one to which only 3 μl of ethanol was added was prepared and cultured under the same conditions.

(Cell recovery and lipid extraction)
The supernatant was removed by centrifugation (4.degree. C., 3,000.times.g, 5 min) of 3 ml of culture medium cultured for 24 hours. After washing with PBS solution, the cells were recovered by centrifugation again to obtain dried cells by lyophilization. Measure 4 mg of dried cells, 300 μl of internal standard (20 μM CG, 10 μM ASG (18: 0-sitosteryl glucoside), TG (12: 0: 12: 0/12: 0), PC (11: 0/11) C: M: 0: 0), LPC (13: 0), PE (12: 0/12: 0), LPE (13: 0), SE (16: 0-d7 cholesterol), d7 cholesterol) C / M = 2/1 And sonicated for 10 minutes. Centrifugation (
The supernatant was recovered at 4 ° C., 15,000 rpm, 5 min). 75 μl of ultrapure water was added and centrifuged (4 ° C., 15,000 rpm, 5 min) to recover the lower layer.

(Measurement of SG · ASG amount by LC-MS analysis)
The entire lower layer collected was placed in an autosampler vial, and 5 μl was subjected to LC-MS. The LC-MS conditions are the same as in Test 2.

(Measurement of SG · ASG content of TG lipase (tgl3, 4, 5) -deficient strain)
A TG lipase (tgl3, tgl4, tgl5) deficient strain was used. The cells precultured in 3 ml of YPD medium were added to 3 ml of YPD medium to an initial concentration of OD ₆₀₀ = 0.02 and cultured with shaking for 24 hours (150 rpm, 30 ° C.). The lipid extraction was performed as described above, and the content of SG · ASG was analyzed by LC-MS.

(AEG analysis of TG lipase deficient strain)
If the TG lipase was an AEG synthetase, it was thought that the AEG amount would be reduced by deleting the TG lipase. Therefore, since budding yeast has Tgl3, Tgl4 and Tgl5 as major TG lipases, the amount of ASG in tgl3, tgl4 and tgl5 deletion strains was measured.

(Result of Test 7)
The results are shown in FIG. As a result of LC-MS analysis, EG and acyl EG (AEG) in which fatty acid was added to EG were not detected in the wild strain, but EG and AEG were both detected in the EG excess accumulating strain (FIG. 15 (a)) ). The type of fatty acid of AEG was 16: 0, 16: 1, 18: 0, 18: 1 (FIG. 15 (b)). In addition, in the EG excess accumulating strain, EG hardly changed depending on the presence or absence of oleic acid (FIG. 15 (a)), but AEG tended to increase by addition of oleic acid (FIG. 15 (b)). From these results, it was suggested that an increase in the amount of TG in budding yeast is associated with an increase in the amount of AEG, and that budding yeast has an AEG synthetase.

  In addition, as a result of LC-MS analysis, the amount of AEG tended to decrease when tgl3, tgl4 and tgl5 were deleted (FIG. 15 (c)). This result supports hypothesis 1 that TG lipase transfers fatty acid to SG.

<Summary of Tests 1 to 7>
(1. Determination of structure of unknown glycolipid)
The unknown glycolipid was eluted with chloroform: acetone = 80: 20.
Analysis of unknown glycolipids by LC-MS, weak alkaline decomposition, β-glucosidase treatment shows that the unknown glycolipid is an acylsteryl glucoside (ASG) in which a fatty acid such as DHA or palmitic acid is esterified to glucose of steryl glucoside (SG) )Met.
Acylsteryl glucoside (ASG) in which DHA was added to steryl glucoside (SG) was a novel structure (FIG. 16).

(2. Comparison of SG and ASG amounts at culture stage)
Acylsteryl glucoside (ASG) increased in the logarithmic growth phase and decreased after the stationary phase. On the other hand, steryl glucoside (SG) tended to increase after glucose depletion.
It was suggested that the acyl steryl glucoside (ASG) may act as a reservoir for DHA accumulation as well as neutral lipids and phospholipids (FIG. 17).

<Discussion of examination>
With regard to glycolipids of labyrinthulas, structural determination and elucidation of biological functions have been advanced for SG. However, unknown glycolipids of unknown structure exist in Labyrinthula separately from SG, and in this study, we aimed at the structural determination of this unknown glycolipid and elucidation of synthetic enzymes.

  As a result of the structural determination of the unknown glycolipid, the unknown glycolipid was ASG in which DHA or palmitic acid was added to SG (FIG. 4B, FIG. 5B, FIG. 6, FIG. 7). ASG has not been reported so far in labyrinthulas, and the structure to which DHA has been added is a novel structure so far not reported in other organisms.

  Labyrinthulas are characterized by the accumulation of highly unsaturated fatty acids such as DHA as TG and SE in oil droplets. As a result of measuring SG and ASG amounts at the culture stage in this test, SG level hardly changed with culture stage, but ASG increased in the logarithmic growth phase and decreased after entering the stationary phase. (FIG. 12, FIG. 13). From this, it is possible that ASG acts as a reservoir of DHA as TG and SE.

  DHA is known to have anti-inflammatory effects, but it is reported that ASG contained in rice bran extract oil has a role to lower elevated blood LDL-cholesterol. ASG bound is applicable to humans as a health food or supplement, taking advantage of not only DHA functionality but also as ASG functionality.

  There are at least four types of SG of Labyrinthula (FIG. 1). So far, SG has been reported to increase under heat stress in fungi and human fibroblasts. SG also increases with heat stress in labyrinthulas, suggesting that SG plays an important role in the development of the environment adaptation mechanism possessed by labyrinthulas, and ASG was similarly related to the environment adaptation mechanism as well. There is a possibility of having a function.

<Test 8: Degradation of A. limacinum-derived DHA-GL-C by lipase>
(Method of test 8)
DHA-GL-C and 25 U of porcine pancreatic lipase were mixed, reacted with a thermomixer at 37 ° C., 750 rpm, and quenched by adding C / M = 2: 1.
(A) The decomposition product reacted with lipase for 28 hours is loaded on TLC, developed with C / M / W = 65/16/2, and is dissolved with copper sulfate reagent (3% copper sulfate dissolved in 15% phosphoric acid solution) Color was developed (lanes boxed in FIG. 18). Triacylglycerol (TG) was digested with the same amount of lipase as a control (right two lanes).
(B) After the time of reaction, the reaction product was quantified by LC-MS.

(Result of Test 8)
The results are shown in FIGS. 18 and 19.
(A) DHA-GL-C was degraded by lipase to release DHA and SG (GL-C) (FIG. 18, +: added lipase,-not added).
(B) The decomposition of DHA-GL-C by lipase was dependent on the reaction time, and DHA-GL-C decreased and free SG (GL-C) increased with the passage of reaction time (FIG. 19).

<Test 9 A. limacinum-derived DHA-GL-C and oxidation measurement of DHA>
(Method of Test 9)
Add 100 μl of 50 mM phosphate buffer pH 7.3 to DHA-GL-C or DHA (5 nmol) purified from 1 mg dry cell weight, and with the container lid open UV irradiation (254 nm, 51 μW / cm ² , from a lamp) 35 cm to the sample, 23 ° C.).
After the irradiation, 200 μl of C / M = 2/1 was added to extract lipids, and DHA-GL-C and DHA were quantified by LC-MS.

(Result of examination 9)
The results are shown in FIG. DHA was oxidized about 50% in 1 hour of UV irradiation, but DHA-GL-C was not oxidized at all in 3 hours of irradiation. Therefore, it was shown that DHA-GL-C was more resistant to oxidation than DHA.

  As a result of the oxidation test by UV irradiation, ASG in which DHA is esterified exists in a stable form in the body, and when it is degraded by lipase, it is converted to functional metabolites such as resolvin and protectin like free DHA. It turned out that there is a possibility.

Claims (18)

Acylsteryl glucoside represented by the following structural formula (I), structural formula (II) or structural formula (III).
Labyrinthula preparation process to prepare a labyrinthula,
Separating the acylsteryl glucoside fraction containing the acylsteryl glucoside from the Labyrinthula;
A purification step of purifying the acylsteryl glucoside from the acyl steryl glucoside fraction separated in the separation step;
A method for producing an acylsteryl glucoside characterized in that   The method for producing an acylsteryl glucoside according to claim 2, wherein the acyl steryl glucoside is a steryl glucoside esterified by docosahexaenoic acid or palmitic acid.   The method for producing an acylsteryl glucoside according to claim 3, wherein the acylsteryl glucoside is a steryl glucoside esterified with docosahexaenoic acid. 5. The acylsteryl glucoside according to claim 4, wherein the acylsteryl glucoside is one or more kinds represented by the following structural formula (I), structural formula (II) or structural formula (III). Production method.
Contains acylsteryl glucoside from Labyrinthula,
A composition characterized by being a composition selected from a food composition, a pharmaceutical composition, and a cosmetic composition.   The composition according to claim 6, wherein the acyl steryl glucoside is esterified by docosahexaenoic acid or palmitic acid to steryl glucoside.   The composition according to claim 7, wherein the acyl steryl glucoside is esterified by docosahexaenoic acid to steryl glucoside. The composition according to claim 8, wherein the acylsteryl glucoside is at least one represented by the following structural formula (I), structural formula (II) or structural formula (III).
  The composition according to any one of claims 6 to 9, wherein the composition is a food composition.   The composition according to any one of claims 6 to 9, wherein the composition is a pharmaceutical composition.   The composition according to any one of claims 6 to 9, wherein the composition is a cosmetic composition.   An antioxidant comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient.   A hair growth or growth agent comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient.   A hypolipidemic agent comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient.   An anti-obesity agent comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient.   An antitumor agent comprising an acylsteryl glucoside derived from labyrinthula as an active ingredient. An agent for the prophylaxis or treatment of atherosclerosis, which comprises an acylsteryl glucoside derived from labyrinthula as an active ingredient.