WO2023157948A1 - Oral composition - Google Patents

Abstract

The present invention addresses the problem of providing a novel lipid functional ingredient that can be taken or administered orally. An example of a composition containing said ingredient is a composition containing glycerol-1-phosphate moieties (CH2OH-CHOH-CH2-OPO2 --), ethanolamine phosphate moieties (NH3 +-CH2-CH2-OPO2 --), and eicosapentaenoic acid moieties CH3CH2(CH=CHCH2)5(CH2)2CO-), the molar ratio of ethanolamine phosphate moieties to glycerol-1-phosphate moieties being 0.01-100.

Description

oral composition

The present invention relates to compositions that are orally ingested or administered. The present invention also relates to methods for increasing muscle or strength, inhibiting muscle or strength loss, improving glucose metabolism, inhibiting fat accumulation in the liver, and improving the balance of omega-3 and omega-6 fatty acids in the body.

Lipids are one of the three major nutrients, and while they occupy an important position as an energy source, it is known that certain lipids have excellent physiological activity.

Fish oil and the like contain a large amount of eicosapentaenoic acid (EPA; (5Z, 8Z, 11Z, 14Z, 17Z)-Eicosapentaenoic acid) in triglycerides. EPA is one of the omega-3 fatty acids, which is essential for fetal development, lowers blood neutral fat (triglyceride), reduces the risk of cardiovascular disease, contributes to the prevention of Alzheimer's disease, etc. has been reported (Non-Patent Document 1).

EPA is also reported to have the effect of suppressing the production of inflammatory lipid mediators such as prostaglandins and leukotrienes from the omega-6 fatty acid arachidonic acid. Furthermore, it is known that EPA itself becomes an anti-inflammatory lipid mediator through metabolism and suppresses inflammation.

Krill oil, an edible oil extracted and refined from Antarctic krill, is rich in phosphatidylcholine, a type of phospholipid. Phosphatidylcholine is known to contribute to memory maintenance. Further, Patent Document 1 describes a cerebral atrophy inhibitor containing, as an active ingredient, a phospholipid containing a highly unsaturated fatty acid as a constituent fatty acid.

However, due to the growing awareness of health among people, there is still a need for new health functional ingredients that can be easily ingested or administered orally.

JP 2013-216654 A

The subject of the present invention relates to the provision of novel functional lipid components that can be orally ingested or administered.

The present inventors have made intensive studies to solve the above-mentioned problems, and as a result, found that glycerol-1-phosphate moiety (CH ₂ OH--CHOH--CH ₂ --OPO ₂ ^-- ), ethanolamine phosphate moiety (NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ ^- -) and eicosapentaenoic acid moieties (CH ₃ CH ₂ (CH=CHCH ₂ ) ₅ (CH ₂ ) ₂ CO-) and glycerol-1-phosphate moieties The inventors have found that compositions having a molar ratio of ethanolamine phosphate moieties to 0.01 to 100 can provide unique physiological activities when orally ingested or administered to a subject, thus completing the present invention.

That is, the present invention includes the following aspects.
[1]
glycerol-1-phosphate moiety (CH ₂ OH-CHOH-CH ₂ -OPO ₂ ^- -), ethanolamine phosphate moiety (NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ ^- -), and eicosapentaenoic acid moiety (CH ₃ CH ₂ (CH═CHCH ₂ ) ₅ (CH ₂ ) ₂ CO—),
the molar ratio of ethanolamine phosphate moieties to glycerol-1-phosphate moieties is from 0.01 to 100;
Compositions taken or administered orally.
[2]
The composition of [1], wherein the molar ratio of said eicosapentaenoic acid moieties to the sum of said glycerol-1-phosphate moieties and ethanolamine phosphate moieties is from 0.01 to 20.
[3]
Phospholipid represented by formula (I)

(wherein R ₁ and R ₂ are selected from hydrogen atoms and acyl groups,
at least one of R ₁ and R ₂ is an eicosapentaenoic acid moiety;
X is selected from glycerol-1-phosphate moieties and ethanolamine phosphate moieties).
[4]
The composition according to any one of [1] to [3], which contains an extract of Shewanella bacteria or a mutant strain thereof.
[5]
containing an extract of Shewanella woodyi or a variant thereof,
The composition according to [4].
[6]
The composition according to [5], which contains an extract of Shewanella woodyi strain GS001 (accession number NITE BP-03460) or a mutant strain thereof.
[7]
For at least one selected from the group consisting of increasing muscle or muscle strength, suppressing muscle or muscle strength loss, improving glucose metabolism, suppressing fat accumulation in the liver, and improving the balance of omega-3 fatty acids and omega-6 fatty acids in the body. The composition according to any one of [1] to [6], which is used.
[8]
Any one of [1] to [6], which is used for the prevention or improvement of at least one selected from the group consisting of diabetes, fatty liver, inflammation, locomotive syndrome, sarcopenia, lifestyle-related diseases and metabolic syndrome The described composition.
[9]
The composition according to any one of [1] to [6], which is used for increasing at least one selected from DHA and DPA in the body.

[10]
For increasing muscle or muscle strength, suppressing muscle or muscle strength loss, improving glucose metabolism, suppressing fat accumulation in the liver, improving the balance of omega-3 fatty acids and omega-6 fatty acids in the body, or diabetes, fatty liver, inflammation , locomotive syndrome, sarcopenia, or metabolic syndrome. or ingesting.

[11]
For at least one selected from the group consisting of increasing muscle or muscle strength, suppressing muscle or muscle strength loss, improving glucose metabolism, suppressing fat accumulation in the liver, and improving the balance of omega-3 fatty acids and omega-6 fatty acids in the body. Use of the composition according to any one of [1] to [6] for the manufacture of an orally ingested or administered formulation.
[12]
For the manufacture of orally ingested or administered formulations used for the prevention or improvement of at least one selected from the group consisting of diabetes, fatty liver, inflammation, locomotive syndrome, sarcopenia, lifestyle-related diseases and metabolic syndrome Use of the composition according to any one of [1] to [6].

[13]
The composition according to any one of [1] to [6], which is used for maintaining or promoting expression of ELOVL2, ELOVL5, FADS1 or FADS2 gene in vivo.
[14]
A method for maintaining or promoting ELOVL2, ELOVL5, FADS1 or FADS2 gene expression in the body, comprising an effective amount of the composition according to any one of [1] to [6]. A method comprising administering to or ingesting by a subject.
[15]
The method according to any one of [1] to [6] for the manufacture of an orally ingested or administered formulation, which is used to maintain or promote the expression of ELOVL2, ELOVL5, FADS1 or FADS2 gene in the body. Use of the composition.

The composition of the present invention can be ingested or administered orally. And, the composition of the present invention can be used for prevention or amelioration of diseases, symptoms or health conditions of subjects due to its unique physiological activity.

1 is a graph showing fatty acid composition in SW oil. 1 is a graph representing the fatty acid composition of ethanolamine-type phospholipids (PE) and glycerol-1-phosphate-type phospholipids (PG) fractions of SW oil. 1 is a flowchart of statistical analysis of Test Examples 1 to 3. FIG. 4 is a graph showing changes in average food intake of mice in each group in Test Example 1. FIG. 2 is a graph showing the gastrocnemius muscle weight of mice in each group after 6-week feeding in Test Example 1. FIG. 2 is a graph showing fasting blood glucose levels of mice in each group on day 41 of feeding in Test Example 1. FIG. 2 is a graph showing blood glucose levels of mice of each group on day 42 of breeding in Test Example 1, 2 hours after feeding. 4 is a graph showing HbA1c values of mice in each group on day 42 of breeding in Test Example 1. FIG. 4 is a graph showing liver weights of mice in each group after feeding for 6 weeks in Test Example 1. FIG. 4 is a graph showing blood triglyceride levels of mice in each group after 6-week feeding in Test Example 1. FIG. Graph showing blood total cholesterol level (T-CHO), LDL cholesterol level (LDL-CHO) and non-HDL cholesterol level (Non-HDL-CHO) in mice of each group after feeding for 6 weeks in Test Example 1. be. 1 is a graph showing the weight of perirenal and posterior abdominal wall fat and the total weight of individual white adipose tissue of mice in each group after feeding for 6 weeks in Test Example 1. FIG. 1 is a graph showing the fatty acid composition of n-3 polyunsaturated fatty acids (PUFA) in livers of mice of each group after feeding for 6 weeks in Test Example 1. FIG. 4 is a graph showing the arachidonic acid (AA) content in the liver of mice of each group after four weeks of feeding in Test Example 2. FIG. 4 is a graph showing the eicosapentaenoic acid (EPA) content in the liver of mice of each group after 4-week feeding in Test Example 2. FIG. 4 is a graph showing the content of docosapentaenoic acid (DPA) in the liver of mice of each group after feeding for 4 weeks in Test Example 2. FIG. 4 is a graph showing the content of docosahexaenoic acid (DHA) in the liver of mice of each group after 4-week feeding in Test Example 2. FIG. 3 is a graph showing the expression level of ELOVL2 in the liver of mice of each group in Test Example 3. FIG. 4 is a graph showing the expression level of ELOVL5 in the liver of mice of each group in Test Example 3. FIG. 4 is a graph showing the expression level of FADS1 in the liver of mice of each group in Test Example 3. FIG. 3 is a graph showing the expression level of FADS2 in the liver of mice of each group in Test Example 3. FIG.

A preferred embodiment of the present invention will be described in detail below. However, the present invention is not limited to the following embodiments. In this specification, the upper limit and lower limit individually described can be arbitrarily combined.

As used herein, "saturated fatty acid" refers to a fatty acid that does not contain a carbon-carbon double bond. "Unsaturated fatty acid" also refers to a fatty acid containing at least one carbon-carbon double bond.

In this specification, fatty acids are sometimes expressed in the form of "total number of carbon atoms: number of double bonds n-X", or in the form with C attached to the head. "nX" means that the double bond first appears at the Xth carbon counted from the methyl end of the carbon chain of the fatty acid. For example, α-linolenic acid is represented as "18:3n-3" or "C18:3n-3". When the part of "nX" is not described, it represents a fatty acid in which only the total number of carbon atoms and the number of double bonds are specified.

As used herein, the "acyl group" corresponding to a specific fatty acid is a portion of the carboxyl group of the fatty acid excluding the OH portion, and is represented by the general formula R-CO- (R is an aliphatic hydrocarbon group). be.

As used herein, "phospholipid" is a generic term for glycerophospholipids and sphingophospholipids. Glycerophospholipids are glycerophospholipids in which one of the three hydroxyl groups of glycerin is phosphorylated. Of these, those in which both remaining hydroxyl groups are acylated are diacyl-type glycerophospholipids, and one of the remaining hydroxyl groups is acylated. Those only acylated are called lysophospholipids.
In this specification, ethanolamine-type phospholipids (PE) and glycerol-1-phosphate-type phospholipids (PG) are ethanolamine phosphate and glycerol-1-phosphate, respectively, with a phosphate ester bond to the hydroxyl group at the 1-position. represents the phospholipids that
In the present specification, phosphatidylethanolamine and phosphatidylglycerol represent diacyl glycerophospholipids containing ethanolamine phosphate and glycerol-1-phosphate as constituents, respectively.

As used herein, "ameliorating" a disease, condition or condition means ameliorating or alleviating the disease, condition or condition; preventing or slowing deterioration of the disease, condition or condition; or reversing the progression of the disease or condition. , stands for prevention or delay.

As used herein, "prevention" of a disease, condition or condition means delaying or preventing the occurrence of the disease, condition or condition; reducing the risk of developing the disease, condition or condition; Represents a reduction in the rate of symptom incidence.

[Compositions Taken or Administered Orally]
The composition of the invention is orally ingested or administered,
glycerol-1-phosphate moiety (CH ₂ OH-CHOH-CH ₂ -OPO ₂ ^- -), ethanolamine phosphate moiety (NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ ^- -) and eicosapentaenoic acid moiety ( containing CH ₃ CH ₂ (CH═CHCH ₂ ) ₅ (CH ₂ ) ₂ CO—),
The molar ratio of ethanolamine phosphate moieties to glycerol-1-phosphate moieties is 0.01-100.

In this specification, a composition that can be orally ingested or administered may be referred to as an "oral composition".

As used herein, the term "glycerol-1-phosphate moiety" refers to a moiety of glycerol-1-phosphate excluding the hydrogen atom of the phosphate group, and has the structural formula CH ₂ OH--CHOH--CH ₂ --OPO ₂ ^--. - is represented. A hydrogen ion or other cation may be associated with the phosphate group (OPO ₂ ⁻ ) of the glycerol-1-phosphate moiety.
As used herein, the term “ethanolamine phosphate moiety” refers to a portion of the phosphate group of ethanolamine phosphate from which hydrogen atoms are removed, represented by the structural formula NH ₃ ⁺ —CH ₂ —CH ₂ —OPO ₂ ^— . be done. A hydrogen ion or other cation may be associated with the phosphate group of the ethanolamine phosphate moiety.
Here, the state in which a hydrogen ion associates with the phosphate group of the glycerol-1-phosphate moiety or the ethanolamine phosphate moiety is CH ₂ OH--CHOH--CH ₂ --OPO ₂ H-- or NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ H-, including a state in which hydrogen is bonded to a phosphoric acid moiety to form a hydroxyl group.
Examples of the other cations include alkali metal ions (sodium ion, potassium ion, etc.) and alkaline earth metal ions (magnesium ion, calcium ion, etc.).

As used herein, the term “eicosapentaenoic acid moiety” refers to an acyl group moiety excluding the OH moiety of the carboxy group of eicosapentaenoic acid, and has the structural formula CH ₃ CH ₂ (CH=CHCH ₂ ) ₅ (CH ₂ ) ₂ CO - is represented.

In the oral composition of the present invention, the molar ratio of ethanolamine phosphate moieties to glycerol-1-phosphate moieties (substance amount of ethanolamine phosphate moieties/substance amount of glycerol-1-phosphate moieties) is From the viewpoint of exhibiting the effect remarkably, it is 0.01 or more, preferably 0.1 or more, more preferably 0.5 or more, still more preferably 1 or more, and even more preferably 1.5 or more. In addition, the molar ratio of the ethanolamine phosphate moiety to the glycerol-1-phosphate moiety is 100 or less, preferably 50 or less, more preferably 20 or less, and still more preferably, from the viewpoint of significantly exhibiting the effects of the present invention. 10 or less, and even more preferably 5 or less.

Molar ratio of the eicosapentaenoic acid moiety to the sum of the glycerol-1-phosphate moiety and the ethanolamine phosphate moiety [(substance amount of eicosapentaenoic acid moiety)/(substance amount of glycerol-1-phosphate moiety + ethanolamine is not particularly limited, but is preferably 0.01 or more, more preferably 0.1 or more, still more preferably 0.2 or more, and further preferably More preferably, it is 0.4 or more. The molar ratio of the eicosapentaenoic acid moiety to the total of the glycerol-1-phosphate moiety and the ethanolamine phosphate moiety is, for example, 20 or less, 10 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 .5 or less.

The content of the eicosapentaenoic acid moiety in the oral composition of the present invention is, for example, 0.01% by mass or more, 0.1% by mass or more, 1% by mass or more, or 4% by mass, relative to the total amount of the oral composition. and may be 50% or less, 40% or less, or 30% or less by weight.

In the oral composition of the present invention, the content of C15:0 fatty acid relative to 100 parts by mass of eicosapentaenoic acid in the total lipid is preferably 5 parts by mass or more, more It is preferably 10 parts by mass or more, more preferably 15 parts by mass or more, and preferably 100 parts by mass or less, more preferably 70 parts by mass or less, and still more preferably 50 parts by mass or less.

In the composition for oral use of the present invention, the content of C16:1 fatty acid relative to 100 parts by mass of eicosapentaenoic acid in the total lipid is preferably 10 parts by mass or more, more It is preferably 15 parts by mass or more, preferably 300 parts by mass or less, more preferably 200 parts by mass or less.

In the composition for oral use of the present invention, the content of docosahexaenoic acid relative to 100 parts by mass of eicosapentaenoic acid in the total lipid is preferably 10 parts by mass or less, more preferably 5 parts by mass or less, and even more preferably 1 part by mass or less. be.

In this specification, when extraction of total lipids from the composition for oral use of the present invention is necessary, as long as there is no problem in the analysis, it is described in the section <<method for fractionating total lipids>> in the Examples. method shall be used. If it is difficult to fractionate by this method, follow the lipid extraction method described in the appendix of the Food Labeling Standards (Cabinet Office Ordinance No. 10, 2015). In the present specification, the analysis of fatty acids in total lipids is performed by the method described in the section [Method for analyzing fatty acids] in Examples.

In the composition for oral use of the present invention, the glycerol-1-phosphate moiety, the ethanolamine phosphate moiety and the eicosapentaenoic acid moiety may constitute molecules different from each other. Alternatively, said glycerol-1-phosphate moiety or said ethanolamine phosphate moiety may be present in the same molecule as said eicosapentaenoic acid moiety. The oral composition of the present invention contains a molecule containing a glycerol-1-phosphate moiety and an eicosapentaenoic acid moiety, or a molecule containing an ethanolamine phosphate moiety and an eicosapentaenoic acid moiety, from the viewpoint of significantly exhibiting the effects of the present invention. It preferably contains at least and more preferably contains a molecule containing a glycerol-1-phosphate moiety and an eicosapentaenoic acid moiety and a molecule containing an ethanolamine phosphate moiety and an eicosapentaenoic acid moiety.

As the molecule containing a glycerol-1-phosphate moiety and an eicosapentaenoic acid moiety, or the molecule containing an ethanolamine phosphate moiety and an eicosapentaenoic acid moiety, a phospholipid represented by the following formula (I) is preferable. Mentioned are:

(wherein R ₁ and R ₂ are selected from hydrogen atoms and acyl groups,
at least one of R ₁ and R ₂ is an eicosapentaenoic acid moiety;
X is selected from glycerol-1-phosphate moieties and ethanolamine phosphate moieties).

The oral composition of the present invention preferably contains both a compound in which X is a glycerol-1-phosphate moiety and a compound in which X is an ethanolamine phosphate moiety in the formula (I).

In the present specification, in the above formula (I), X represents glycerol-1-phosphate moieties as EPA-bound glycerol-1-phosphate-type phospholipids (EPA-PG), and X represents ethanolamine phosphate moieties. Sometimes referred to as EPA-bound ethanolamine-type phospholipids (EPA-PE).

In the composition for oral use of the present invention, the content of the EPA-bound ethanolamine-type phospholipid relative to 1 part by weight of the EPA-bound glycerol-1-phosphate-type phospholipid is 0.01 part by weight or more, preferably 0.01 part by weight. It is 1 part by mass or more, more preferably 0.5 parts by mass or more, still more preferably 1 part by mass or more, and even more preferably 1.5 parts by mass or more. Then, the content of the EPA-bound ethanolamine-type phospholipid with respect to 1 part by weight of the EPA-bound glycerol-1-phosphate-type phospholipid is 100 parts by weight or less, preferably, from the viewpoint of significantly exhibiting the effects of the present invention. It is 50 parts by mass or less, more preferably 20 parts by mass or less, still more preferably 10 parts by mass or less, and even more preferably 5 parts by mass or less.

The total carbon number of the acyl group of formula (I) is, for example, 3-35, preferably 9-31, more preferably 14-24.

The number of double bonds in the acyl group of formula (I) is, for example, 0-10, preferably 0-6, more preferably 0-5.

The acyl groups of R ₁ and R ₂ in the formula (I) are preferably acyl groups contained in phospholipids in organisms, more preferably C14:0 (myristic acid), C14:1, C15:0 ( pentadecanoic acid), C16:0 (palmitic acid), C16:1, C17:1, C18:0 (stearic acid), C18:1, C18:2, C18:3n-3 (α-linolenic acid), C18: 3n-6 (γ-linolenic acid), C19:0, C20:0 (arachidic acid), C20: 1n-9 (cis-11-eicosenoic acid), C20:2n-6, C20:3n-3, C20: 3n-6 (dihomo-γ-linolenic acid), C20: 4n-6 (arachidonic acid; AA), C20: 5n-3 (eicosapentaenoic acid; EPA), C22:0 (behenic acid), C22: 1n-9 (erucic acid), C22:4n-6, C22:6n-3 (docosahexaenoic acid; DHA), C23:0, C24:0 (lignoceric acid) and C24:1n-9 (nervonic acid) selected, more preferably C12:0, C13:0, C14:0, C15:0, C16:0, C16:1, C17:1, C18:0, C18:1, C19:0, C20:3n -3 and acyl groups corresponding to C20:5n-3.

In the oral composition of the present invention, the total content of the phospholipids represented by the formula (I) is appropriately adjusted depending on the form, composition, purpose of use, usage, dosage, etc. of the oral composition. The total content of phospholipids represented by formula (I) is, for example, 0.01% by mass or more, 0.1% by mass or more, 1% by mass or more, 10% by mass or more, relative to the total amount of the oral composition. , 20% by weight or more, or 50% by weight or more, and may be, for example, 100% by weight or less, 80% by weight or less, or 70% by weight or less.

The glycerol-1-phosphate moiety, the ethanolamine phosphate moiety and the eicosapentaenoic acid moiety of the oral composition of the present invention, and the phospholipid represented by the formula (I) may be derived from organisms. It may be synthesized, or it may be derived from a mixture of two or more of them, but from the viewpoint of safety in oral ingestion, preferably. Said organisms are, for example, selected from microorganisms (algae, bacteria, archaea, fungi, yeast, etc.), plants, marine and aquatic animals (eg, fish, crustaceans, etc.), and terrestrial animals.

The oral composition of the present invention preferably contains an extract of a bacterium of the genus Shewanella or a mutant strain thereof, from the viewpoint of exhibiting the effects of the present invention more remarkably. Bacteria belonging to the genus Shewanella belong to the family Shewanellaceae, and are Gram-negative, non-spore-forming anaerobic bacilli that are mainly distributed in the marine environment, and include barophilic bacteria, psychrophilic bacteria, and the like that live in the deep sea and the like.

As used herein, a "mutant strain" of a specific species refers to a strain in which a mutation has occurred in the DNA possessed by the original strain of the species. The mutation is not particularly limited, and may be, for example, base substitution, deletion, insertion, duplication, translocation or inversion in genomic DNA or a plasmid originally carried by the strain. Mutations may be naturally occurring or artificially occurring.

The Shewanella bacterium or its mutant may further contain naturally-derived or artificial DNA such as plasmids and bacterial artificial chromosomes (BAC) in addition to genomic DNA.

The genome sequence of the mutant is, for example, 90% or more, 95% or more, 97% or more, 98% or more, 99% or more, 99.5% or more, 99.9% or more of the reference genome sequence of the organism. , has a sequence identity of greater than or equal to 99.99% or greater than or equal to 99.999%.

Examples of the Shewanella bacteria include Shewanella woodyi, Shewanella electrodiphila, S. marintestina, S. schlegeliana, and Shewanella cylae. (S. sairae), S. pealeana, S. hanedai, S. gelidimarina, S. pneumatophore, Shewanella baltica (S. baltica), S. halifaxensis, S. kaireitica, S. glacialipiscicola, S. fidelis, Shewanella aquimarina (S. aquimarina), S. waksmanii, S. marisflavi, S. affinis, S. colwelliana, S. violacea, Shewanella・S.benthica, S.sediminis, etc. Among them, the Shewanella bacterium is preferably Shewanella woodyi, more preferably Shewanella woodyi GS001 strain. For information on Shewanella udii, see, for example, Brenner et al. (The Proteobacteria, 2005, p.480-491), Zhao et al. (Int. J. Syst. Evol Microbiol., 2005, Vol. 55, p. 1511-1520), Makemson, J.C. et al. (Int. J. Syst. Bacteriol., 1997, Vol.47, Issue 4, p.1034-1039), Ivanova et al. (Int. J. Syst. Evol. Microbiol. 2003, Vol.53, p.577-582).

Alternatively, in the Shewanella bacterium, the sequence identity of the 16S rDNA sequence thereof to the 16S rDNA sequence (SEQ ID NO: 1) of the following Shewanella oudii strain GS001 is preferably 95% or more, more preferably 96% or more, and 97%. 98% or more is more preferable, and 99% or more is particularly preferable. Here, the identity of the 16S rDNA sequence is determined using Align two or more sequences in blastn (for base sequences) of BLAST (https://blast.ncbi.nlm.nih.gov/) of NCBI. It is the percentage of "Identities" obtained by inputting a sequence to compare a specific sequence to Query Sequence and performing alignment with default parameters.

The Shewanella udii strain GS001 (hereinafter sometimes referred to as "GS001 strain") is a strain isolated from the intestinal tract of Nigisu, a deep-sea fish. The 16S rDNA sequence of strain GS001 is shown in SEQ ID NO:1.

The GS001 strain is characterized by being able to grow well using glucose as the sole carbon source. For example, the GS001 strain was cultured with shaking (150 rpm) at 15°C using a 100 mL flask containing 50 mL of a modified M9 liquid medium containing 10% (w/v) glucose (modified M9 + Glc liquid medium) for 48 hours. , OD600nm can grow to a cell density of 5 or more.
The GS001 strain is characterized by being able to grow well in a semi-synthetic medium containing glucose and yeast extract. For example, the GS001 strain is aerated cultured at 15 ° C. (1.0 vvm air, 150 to 750 rpm ), and when a glucose solution as a carbon source and an ammonium aqueous solution as a nitrogen source are fed at appropriate rates, respectively, the dry cell weight can grow to 35 g/L or more in 75 hours.
The GS001 strain is characterized by a high EPA content. For example, the GS001 strain was aerated at 15° C. (1 vvm air, 150-750 rpm) using a 3 L jar fermenter containing 2 L of modified M9+Glc liquid medium containing 0.5% (w/v) yeast extract. At times, the EPA content per dry cell weight can be maintained above 0.5 g/100 g dry cell weight.
The GS001 strain is characterized by being able to grow in sodium chloride concentrations as low as 0.5% (w/v). For example, the GS001 strain was cultured with shaking (150 rpm) at 15 ° C. using a 100 mL flask containing 50 mL of modified M9 + Glc medium with a sodium chloride concentration of 0.5% (w / v). can grow to a density of 3 or more.

The GS001 strain was granted accession number NITE P-03460 on April 15, 2021 at the Patent Microorganism Depositary Center, National Institute of Technology and Evaluation (zip code 292-0818, 2-Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan). 5-8 Room 122).
The GS001 strain is identified as Shewanella woodyi, accession number NITE BP-03460, and is registered at the Patent Microorganism Depositary Center, National Institute of Technology and Evaluation (zip code 292-0818, 2-5 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan). 8 122) under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure (National Deposit Date April 15, 2021; Request for Transfer to International Deposit Date April 4, 2022 Day).

The genus Shewanella or its mutants contain the phospholipid of formula (I) at a high intracellular content. Therefore, an extract of the genus Shewanella or a mutant strain thereof can be suitably used in the oral composition of the present invention. And, since the Shewanella udii GS001 strain has the characteristics described above, it is particularly suitable for industrial production of the phospholipid of the formula (I).

Bacteria of the genus Shewanella and their mutant strains can be cultured using media commonly used for culturing heterotrophic bacteria. Culture conditions are not particularly limited as long as Shewanella bacteria can grow. ), and Shewanella electrodyphila can be set based on the description in Zhang J. et al. (PLoS One, 2017, Vol.12, No.11:e0188081).

The extract of the genus Shewanella or its mutants is not particularly limited as long as it is an extract of cell components, for example, a cell disruption obtained by disrupting cells, a cell lysate obtained by lysing cells, or any of them may be a solvent extract obtained by solvent extraction from Among them, the extract of the genus Shewanella or its mutant is preferably a solvent extract from the viewpoint of improving the absorbability of the active ingredient into the body.

The solvent used in the preparation of the solvent extract is not particularly limited as long as it can be used in an oral composition that can be ingested or administered, and is appropriately selected depending on the form of the oral composition (medicine, food and drink, etc.). can choose. Alcohol, for example, is preferred as the solvent.

The extraction of the phospholipid of the formula (I) from Shewanella bacteria or mutants thereof can be carried out by known methods.
For example, cells are recovered from the culture medium of the bacterium of the genus Shewanella or its mutant strain by centrifugation, filtration, or the like, and all lipids are extracted from the cells with the above solvent. The amount of extraction solvent used for cells is not particularly limited. The amount of extraction solvent can be, for example, about 1 to 1000 times (preferably about 5 to 200 times) the amount of cells. The extraction operation can usually be carried out under normal pressure in the range from room temperature to the boiling point of the solvent. The extraction operation may be performed only once or may be performed multiple times. For example, a fresh extraction solvent may be added again to the cell residue that has been subjected to the extraction operation once, or the extraction operation may be performed again. After the extraction operation, if necessary, cell debris may be removed by centrifugation, filtration, ultrafiltration, or the like. Alternatively, the extraction solvent may be removed by heating or distillation under reduced pressure using an evaporator or the like. Further, various purification treatments may be performed to purify the phospholipids. Purification treatments include, for example, salting out, dialysis, recrystallization, reprecipitation, solvent extraction, adsorption, concentration, filtration, gel filtration, ultrafiltration, various chromatography (thin layer chromatography, column chromatography, ion exchange chromatography, high-performance liquid chromatography, adsorption chromatography, etc.), but are not limited thereto.

The oral composition of the present invention is orally ingested or administered. ), pharmaceuticals, quasi-drugs, feeds or feeds, or preparations for their manufacture (eg, additives).

When the oral composition of the present invention is a drug or quasi-drug, its forms include, for example, liquids, tablets, powders, granules, capsules, powders, lozenges, syrups and the like.

When the composition for oral use of the present invention is a food or drink, its form includes, for example, sweets (caramel, candy, etc.), frozen desserts (ice cream, etc.), dairy products (yogurt, etc.), various processed foods, various seasonings. foods, various beverages, functional foods, dietary supplements, and supplements.

The oral compositions of the present invention may optionally contain the above glycerol-1-phosphate moiety (CH ₂ OH--CHOH--CH ₂ --OPO ₂ ^-- ), ethanolamine phosphate moiety (NH ₃ ⁺ --CH ₂ —CH ₂ —OPO ₂ ^— —), or components other than those having an eicosapentaenoic acid moiety (CH ₃ CH ₂ (CH=CHCH ₂ ) ₅ (CH ₂ ) ₂ CO—) (referred to as “other components”) may contain.

When the composition for oral use of the present invention is in the form of a drug or quasi-drug, or a formulation for manufacturing thereof, other ingredients include, for example, pharmaceutically acceptable carriers (excipients, disintegrants, binders , lubricants, colorants, plasticizers, antioxidants, pH adjusters, thickeners, surfactants, stabilizers, preservatives, perfumes, fluidizers, liquid media, foaming agents, etc.) Any one or a combination of two or more selected from, but not limited to, these.

When the composition for oral use of the present invention is in the form of a food or drink, a feed or feed, or a formulation for producing them, other ingredients include, for example, fish meat, vegetables, grains, dairy products, fermented products, spices, and proteins. , amino acids, sugars, various seasonings, sweeteners, flavoring agents, flavors, oils and fats, vitamins, thickeners, gelling agents, antioxidants, preservatives, chelating agents, pH adjusters, colorants Any one or a combination of two or more selected from, but not limited to, these.

The oral compositions of the present invention contain glycerol-1-phosphate moieties (CH ₂ OH-CHOH-CH ₂ -OPO), such as EPA-bound glycerol-1-phosphate-type phospholipids and EPA-bound ethanolamine-type phospholipids. ₂ ^- -) and ethanolamine phosphate moieties (NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ ^- -), and eicosapentaenoic acid moieties (CH ₃ CH ₂ (CH=CHCH ₂ ) ₅ (CH ₂ ) ₂ CO -) as an active ingredient. And, the composition for oral use of the present invention contains an effective amount of the above-mentioned active ingredient, so that, for example, the effect of increasing muscle; action to suppress the rise of; action to suppress fat accumulation in the liver; action to increase omega-3 fatty acids (especially eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid) in the body (e.g., liver) and omega-6 fatty acids (especially, arachidonic acid); reducing blood triglycerides, total cholesterol, LDL cholesterol and non-HDL cholesterol; and reducing the amount of white adipose tissue. These effects of the composition for oral use of the present invention are remarkably higher than krill oil containing a large amount of phosphatidylcholine containing docosahexaenoic acid and eicosapentaenoic acid as fatty acids.

Since the oral composition of the present invention increases omega-3 fatty acids as described above, it can be used to increase omega-3 fatty acids in the body (for example, liver). The omega-3 fatty acid is preferably at least one selected from docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA), more preferably DHA.

As used herein, "improving the balance between omega-3 fatty acids and omega-6 fatty acids" means increasing the ratio of omega-3 fatty acids to omega-6 fatty acids in total lipids in the body (for example, liver). Omega-6 fatty acids include, for example, arachidonic acid (AA), linoleic acid, γ-linolenic acid, dihomo γ-linolenic acid, docosatetraenoic acid and 22:5n-6 docosapentaenoic acid (ospondonic acid). Examples of omega-3 fatty acids include eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), 22:5n-3 docosapentaenoic acid (clupanodonic acid, DPA), α-linolenic acid, stearidonic acid and eicosatetra. enoic acid.

As mentioned above, omega-6 fatty acids are metabolized to pro-inflammatory lipid mediators and omega-3 fatty acids are metabolized to anti-inflammatory lipid mediators. , leading to the prevention or amelioration of inflammation in the body. Therefore, the oral composition of the present invention also has an effect of preventing or improving inflammation.

The oral composition of the present invention also exerts an effect of preventing or ameliorating conditions, symptoms or diseases associated with decreased muscle or muscle strength, such as locomotive syndrome and sarcopenia, through its action of increasing muscle mass.
The composition for oral use of the present invention, through the action of increasing muscle, the action of suppressing the rise of blood sugar level, the action of suppressing the accumulation of fat in the liver, and the action of suppressing the increase of white adipose tissue, is effective against lifestyle-related diseases or metabolic syndrome. There is also an effect of prevention or improvement of

As described above, the oral composition of the present invention, for example, increases muscle or muscle strength, suppresses muscle or muscle strength loss, improves glucose metabolism (for example, improves blood sugar level, more specifically, at least one selected from the group consisting of improvement of hyperglycemia, improvement of postprandial blood sugar level, improvement of average blood sugar level or HbA1c level, etc.), suppression of fat accumulation in the liver, and improvement of the balance of omega-3 fatty acids and omega-6 fatty acids in the body It can be used for one species.
In addition, the composition for oral use of the present invention includes at least one selected from the group consisting of diabetes, fatty liver (non-alcoholic fatty liver or alcoholic fatty liver), inflammation, locomotive syndrome, sarcopenia, lifestyle-related diseases and metabolic syndrome. can be used for the prevention or improvement of

The composition for oral use of the present invention is further selected from the group consisting of improving blood lipid status (for example, improving blood neutral fat (triglyceride) level, cholesterol level or LDL cholesterol level, etc.) and promoting weight loss. can also be used for at least one
The oral composition of the present invention is also used for the prevention or improvement of at least one selected from the group consisting of dyslipidemia (e.g., hypertriglyceridemia, hyperLDL cholesterolemia), arteriosclerosis and obesity. can be used.

The oral composition of the present invention can be used therapeutically or non-therapeutically. As used herein, the term "non-therapeutic" means not including medical practice, for example, not including surgery, treatment or diagnosis of humans by a doctor or those under the direction of a doctor.

The oral composition of the present invention has the effect of maintaining or promoting the expression of the ELOVL2, ELOVL5, FADS1 or FADS2 gene in the liver of a subject, as shown in the Examples. Therefore, the oral composition of the present invention can be used to maintain or promote ELOVL2, ELOVL5, FADS1 or FADS2 gene expression (eg, mRNA expression) in the body (preferably in the liver).

ELOVL2 (Fatty Acid Elongase 2) and ELOVL5 (Fatty Acid Elongase 5) are genes encoding enzymes that catalyze chain elongation of unsaturated fatty acids. FADS1 (Fatty Acid Desaturase 1) and FADS2 (Fatty Acid Desaturase 2) are genes encoding enzymes that introduce double bonds into fatty acids. These genes are involved in the synthesis of polyunsaturated fatty acids (PUFA) such as EPA, DHA and DPA. As shown in the examples, the oral composition of the present invention, unlike krill oil and other oils containing polyunsaturated fatty acids of a different type than the present invention, inhibits the expression of these genes in the liver. does not reduce Although not to be limited to a particular mechanism, differences in the expression of these genes in the liver may be one reason why the oral composition of the present invention increases omega fatty acids in the liver compared to krill oil and the like. It is speculated that it brings about the effect of improving the balance of omega-3 fatty acids and omega-6 fatty acids.

The oral composition of the present invention may be a composition indicating the above uses as efficacy or function. Such indications include, for example, direct indications of the above uses or indications that substantially imply the above uses.

Examples of direct indications for the above uses include “maintenance of muscle/strength” and “keep muscle” as examples of indications for suppressing muscle or muscle strength loss; "Slows blood sugar level", "Lowers blood sugar level", "Reduces high blood sugar level to normal", "Slows postprandial rise in blood sugar level", "Reduces postprandial blood sugar level "Reduce blood sugar rise", "Reduce postprandial blood sugar level", "Reduce high fasting blood sugar level to normal", "Lower blood sugar", etc.; "reduce visceral fat", "reduce visceral fat accumulation", "reduce liver fat", "reduce visceral fat", "reduce belly fat", etc., but are not limited to these.

Examples of indications that substantially indicate the above uses include, for example, figurative indications of the above uses, indications of subjects for which intake or administration is recommended, and indications that they act on specific diseases, symptoms, or health conditions. etc. Examples of such indications include "for those who are concerned about muscle weakness," "muscle/strength support," "muscle storage," "blood sugar care," "blood sugar support," and "for those who are concerned about blood sugar levels." ", "blood sugar countermeasures", "those who are concerned about belly fat", "refreshing around the stomach", "anti-inflammatory", "metabolite countermeasures", "locomo countermeasures", "lifestyle disease countermeasures", etc. include but are not limited to:

Subjects to which the oral composition of the present invention is ingested or administered are humans or non-human animals (e.g. mammals other than humans), preferably diabetes, fatty liver, inflammation, locomotive syndrome, sarcopenia or metabolic syndrome. or at risk of having those diseases, conditions or conditions.

The target age is not particularly limited, but it applies to adults, for example.

The dosage or intake of the oral composition of the present invention is appropriately determined according to the body weight, sex, age, condition, or other factors of the subject of administration or intake.
The dosage or intake of the oral composition of the present invention is, for example, 0 as the total amount of the phosphorylated ethanolamine moiety, the glycerol-1-phosphate moiety and the eicosapentaenoic acid moiety per day. .1-1,000 mg/kg body weight, 0.5-500 mg/kg body weight or 1-100 mg/kg body weight.
Oral compositions of the invention are administered or taken, for example, 1 to 5, 1 to 3, or 2 to 3 times daily.

The present invention will be described in more detail below using examples, but the present invention is not limited to these scopes. Unless otherwise specified, the unit of the amount of components in each table of the test examples below is % by mass. N. D. indicates that the component was not detected in the measurement.

[Analysis method of fatty acid]
In the test examples below, the fatty acid composition in the lipid samples was determined by gas chromatography under the following conditions after the sample lipids were methyl-esterified using a fatty acid methylation kit (Nacalai Tesque, Inc.). .
<Analysis conditions>
Apparatus: GC-2014 Gas Chromatograph (Shimadzu Corporation)
Column: Fused Silica Capillary Column Omegawax320 (30 m×0.32 mm id)
(Supelco, Inc.)
Carrier gas: helium Helium flow rate: 1 mL/min
Column temperature: 200°C
Injection temperature: 250°C
Detector temperature: 260°C
Detector: C-R8A Chromatopac Integrator (Shimadzu Corporation)
<Calculation of fatty acid composition>
Each fatty acid peak in the obtained chromatogram was attributed based on the retention time of each fatty acid standard substance, and the composition of each fatty acid (unit: % by mass) was calculated as a percentage of the total peak area.

[Sample used for sample preparation]
<Shewanella udii GS001 strain extract (SW oil)>
A Shewanella udii GS001 strain extract (hereinafter sometimes referred to as SW oil) was prepared by the following procedure.
(1) Cells collected from a culture of Shewanella udii strain GS001 were dried.
(2) Lipids were extracted from the dried microbial cells using alcohol.
(3) The extract was filtered and the filtrate was further concentrated.
(4) Separation operation was performed using hexane, the hexane fraction was separated and concentrated, and the precipitate was dried to obtain SW oil.

<Shewanella Electrodyphylla Extract (SE Oil)>
A Shewanella electrodyphila extract (hereinafter sometimes referred to as SE oil) was prepared from a culture of Shewanella electrodyphila by the same method as for SW oil.

<Krill oil, fish oil>
Krill oil contains mainly phosphatidylcholine (PC) as phospholipid and little PE and PG. In this test example, a commercial product from Aker BioMarine was used. A fish oil commercially available from Maruha Nichiro Co., Ltd. was used.
<Soybean oil, lard>
Soybean oil used was Fuji Film Wako Pure Chemical Co., Ltd., and lard was used commercially available from Junsei Chemical Co., Ltd.

<Sample analysis results>
Table 1 shows the contents of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), ethanolamine-type phospholipid (PE), and glycerol-1-phosphate-type phospholipid (PG) in SW oil and krill oil. Ta. The contents of EPA and DHA represent the total amount of free fatty acids and those contained as acyl groups in triglycerides, phospholipids, and the like. As shown in Table 1, SW oil does not contain DHA.

In addition, the results of fatty acid composition analysis of SW oil and the polar lipid fraction of SW oil are shown in Figures 1A and 1B. The ratio of EPA to the total amount of fatty acids in the SW oil was 5.42% by mass.

Total lipids were extracted from dried cells of Shewanella electrodyphila by the following procedure, and polar lipids were further fractionated. As a result, the content of phospholipid-type EPA relative to the total amount of EPA in the lipid obtained from the dried cells was 61.3% by mass, the content of neutral lipid-type EPA was 32.6% by mass, and the content of glycolipid-type EPA was The amount was 6.1% by weight. In addition, PG was 32.4% by mass and PE was 59.7% by mass with respect to the total amount of phospholipids in the lipids obtained from the dried cells.
<<Method for fractionating total lipids>>
In the following test examples, extraction of total lipids from samples was performed by the following procedure.
(1) After adding 10 times the amount of alcohol to the sample and extracting the lipid overnight in a dark place, solid-liquid separation was performed by suction filtration, and the filtrate was recovered.
(2) The same amount of alcohol was added to the residue obtained by solid-liquid separation, and the mixture was filtered to collect the filtrate.
(3) The filtrates of (1) and (2) were combined, and the solvent was distilled off from the filtrates using a rotary evaporator.
(4) About 20 g of lipid was weighed into a 500 mL medium bottle, and 800 mL of hexane was added. A medium bottle was placed in an ultrasonic bath, and the lipid was dispersed in hexane.
(5) Add 1 L of alcohol in advance to a separating funnel, mix, add lipid dissolved in hexane, mix and leave overnight to separate into two layers, then remove the upper layer (hexane fraction). Recovered.
(6) The solvent was distilled off from the recovered upper layer using a rotary evaporator, and the obtained lipid-soluble components were used as total lipids.
<<Method for fractionating polar lipids>>
(7)
50 mg of the total lipid obtained was stripped onto a preparative silica gel TLC plate (PLC Silica gel 60 F ₂₅₄ , 1 mm thick; Merck KGaA) and treated with chloroform:methanol:water (65:25:4, v/v/ v) was developed as a developing solvent. After drying in a fume hood, the PE and PG fractions were peeled off from the TLC plate, transferred into a glass column, and each lipid was eluted with 50 mL of methanol. Thereafter, methanol was distilled off using an evaporator to obtain PE and PG.

Table 2 shows the fatty acid composition analysis results in the total lipids of SE oil, fish oil, and krill oil.

〔Statistical analysis〕
In Test Examples 1 to 3 below, the results of statistical analysis are shown as mean±standard error (SE). Statistical analysis was performed according to the flow shown in FIG.

[Test Example 1. Physiological Activity Evaluation Test of Shewanella Bacteria Extract]
<Animal test method>
(1) Diabetes model mice KK-A ^y mice (4 weeks old, male; purchased from Clea Japan, Inc.) were fed with MF feed (manufactured by Oriental Yeast Co., Ltd.) for 1 week, and then supplied. used as a test animal. Control group as test groups (Control; G1), SW oil group (Shewanella; G2) and low dose krill oil group (Krill-L; G3), high dose krill oil group (Krill-H; G4) four groups (each Groups (n=7) were established, and feed and water having the composition shown in Table 3 were administered to each group for 6 weeks. Feed was given ad libitum from the 0th to 8th days after the start of rearing, and from the 9th day onwards, feeding was restricted according to the food intake of the SW oil group (Fig. 3). Water was given ad libitum. There was no significant difference in body weight transition of mice between groups during the study.

(2) After the breeding period, whole blood was collected from the mouse under isoflurane inhalation anesthesia and organs/tissues were removed from the posterior vena cava using a heparinized injection needle (Terumo Corporation) and a syringe (Terumo Corporation). , was used for each of the following evaluations.
(i) Evaluation of gastrocnemius muscle weight: After 6 weeks of feeding, the gastrocnemius muscle of each group was removed and weighed.
(ii) Evaluation of blood glucose level and HbA1c: fasting mouse blood glucose level on day 41 was measured. In addition, the blood sugar level and HbA1c were measured 2 hours after feeding (under non-fasting) on the day of termination of the 6-week feeding (day 42). The blood glucose level was measured by incising the vein at the tip of the tail of the mouse with a scalpel under no anesthesia, and measuring the blood leaking from the incised surface with a blood glucose measuring device (Statstrip Express Glucose Ketone, manufactured by Nova Biomedical Co., Ltd.). The blood glucose was sucked into the connected blood glucose measuring chip and measured. For the HbA1c measurement, 20 μL of the blood obtained by the above whole blood collection was aliquoted and stored at −70° C. or below. A DCA bandage (manufactured by SIEMENS) was used to measure HbA1c.
(iii) Evaluation of liver weight: After 6 weeks of feeding, the liver of each group of mice was excised and weighed.
(iv) Evaluation of blood triglycerides and blood cholesterol: The blood obtained by the above whole blood collection was centrifuged at 3000 rpm for 15 minutes using a table top refrigerated centrifuge 2800 (Kubota Shoji Co., Ltd.), and heparin plasma was removed. Obtained. 200 μL of heparin plasma was collected in advance, and triglyceride (TG), total cholesterol (T-CHO), LDL cholesterol (LDL-CHO) and HDL cholesterol (HDL —CHO) was measured. Non-HDL cholesterol (Non-HDL-CHO) was determined by subtracting the amount of HDL cholesterol from total cholesterol. The heparinized plasma collected in advance was stored at −70° C. or below until the measurement.
(v) Evaluation of adipose tissue weight: White adipose tissue was removed from each group of mice after feeding for 6 weeks, and the weight of the perirenal/retroabdominal fat and all of the white adipose tissue was measured.
(vi) Evaluation of fatty acid composition in liver: n-3 polyunsaturated fatty acid (PUFA) composition in total lipid in the liver of each group of mice after feeding for 6 weeks was measured. Total liver lipids were extracted by the Folch method. After the obtained total lipid was methyl-esterified, the fatty acid composition was analyzed by gas chromatography under the above conditions.

<Results>
(1) Evaluation of gastrocnemius muscle weight FIG. 4 shows the results of comparing the gastrocnemius muscle weights of the mice in each group. The SW oil group significantly increased the gastrocnemius muscle weight compared to the control group, whereas no significant difference was observed between the low dose krill oil group and the high dose krill oil group.

(2) Evaluation of blood sugar level and HbA1c The fasting blood sugar level on the 41st day is shown in FIG. 5A, and the blood sugar level and HbA1c after 2 hours of feeding on the 42nd day are shown in FIGS. 5B and 5C. As for the fasting blood sugar level and the blood sugar level 2 hours after feeding, the blood sugar level was suppressed significantly lower in the SW oil group than in the control group and the two groups administered with krill oil. The HbA1c value was also significantly lower in the SW oil group than in the control group and the two groups to which krill oil was administered.

(3) Evaluation of Liver Weight FIG. 6 shows the results of comparing the liver weights of the mice in each group after completing feeding for 6 weeks. The SW oil group did not show a significant difference from the control group, but the two groups administered krill oil showed a significant increase in liver weight compared to the control group. It is speculated that the phospholipids contained in krill oil induced fat accumulation in the liver.

(4) Evaluation of Blood Triglyceride and Blood Cholesterol The evaluation results of blood triglyceride and cholesterol are shown in FIGS. 7A and 7B. The SW oil group had significantly lower blood triglycerides, total cholesterol, LDL cholesterol and non-HDL cholesterol levels than the control group. On the other hand, there was no significant difference in these values between the two groups administered with krill oil and the control group.

(5) Evaluation of Adipose Tissue Weight FIG. 8 shows the results of comparing the adipose tissue weights of the mice of each group after completing feeding for 6 weeks. In the SW oil group, the total amount of white adipose tissue and the weight of perirenal and posterior abdominal wall fat were significantly lower than in the control group. On the other hand, there was no significant difference in these values between the two groups administered with krill oil and the control group.

(6) Evaluation of Fatty Acid Composition in Liver FIG. 9 shows the n-3 PUFA composition in the liver of each group of mice after feeding for 6 weeks. The SW oil group showed a significant increase in the amount of EPA (20:5n-3) accumulated in the liver compared to the control group, and a significant increase in its proportion in the fatty acid composition was also observed. Furthermore, surprisingly, DHA (22:6n-3) and docosapentaenoic acid (DPA) (22:5n-3), which are hardly contained in SW oil, also account for their accumulated amount and fatty acid composition in the liver. percentage increased significantly. On the other hand, in the krill oil group, although the krill oil contains a relatively large amount of DHA, an increase in the amount of DHA accumulated in the liver and the proportion of DHA in the fatty acid composition was not observed. Also, no significant increase in DPA in the liver was observed.

(7) Summary From the above, the Shewanella bacterium extract has (i) an effect of increasing muscle mass or an effect of suppressing muscle loss, (ii) fasting blood sugar level, postprandial blood sugar level, and average blood sugar level reflected by HbA1c (iii) suppressing fat accumulation in the liver, (iv) suppressing increases in total cholesterol, LDL cholesterol and non-HDL cholesterol, and improving the state of blood lipids. (v) an effect of suppressing an increase in the amount of white adipose tissue; and (vi) an effect of increasing EPA, DPA and DHA in the liver. A comparison of the effects of the SW oil group and the low dose krill oil group shows that these effects are due to EPA-bound ethanolamine-type phospholipids and EPA-bound glycerol-type phospholipids.

[Test Example 2. Physiological Activity Evaluation Test of Shewanella Bacteria Extract]
<Animal test method>
C57BL/6J mice (9 weeks old, male; purchased from Japan Charles River Co., Ltd.) were preliminarily fed with MF feed (Oriental Yeast Co., Ltd.) for 1 week and used as test animals. The following 7 groups (each n=7) were provided as test groups.
G1: control group G2: low dose SE oil group (Shewanella-L)
G3: High dose SE oil group (Shewanella-H)
G4: low dose fish oil group (Fish-L)
G5: High dose fish oil group: (Fish-H)
G6: Low dose krill oil group (Krill-L)
G7: high dose krill oil group (Krill-H)

(2) Mice in each group were bred by freely ingesting the feed and water having the composition shown in Table 4 for 4 weeks. After the breeding period ended, the liver was removed from the mouse.

(3) Total liver lipids were extracted by the Folch method. After the obtained total lipid was methyl-esterified, the fatty acid composition was analyzed by gas chromatography under the above conditions.

<Results>
The results for the liver omega-6 fatty acid arachidonic acid and the omega-3 fatty acids EPA, docosapentaenoic acid (DPA) and DHA are shown in Figures 10A-D. Similar to the fish oil group and the krill oil group, the two groups administered with SE oil showed a decrease in the amount of arachidonic acid compared to the control group. Also, in the two groups to which SE oil was administered, EPA and DPA significantly increased compared to the fish oil group and the krill oil group.

From the above, it was clarified that the Shewanella genus extract has the effect of significantly increasing the ratio of omega-3 fatty acids to omega-6 fatty acids. In comparison with the administration of fish oil and krill oil, it is understood that this effect is also due to EPA-bound ethanolamine-type phospholipids and EPA-bound glycerol-type phospholipids.

[Test Example 3. Verification of Effect of Shewanella Bacterial Extract on Gene Expression]
Total mRNA was extracted from liver samples of mice collected after 6 weeks of breeding in Test Example 1, and cDNA was prepared using reverse transcriptase. The amount of mRNA derived from each gene of ELOVL2, ELOVL5, FADS1 and FADS2 was quantified by qPCR method using GAPDH as a reference gene. As primers for each gene, TaqMan Gene Expression Assay products (manufactured by Applied Biosystems Japan) with the product numbers shown below were used, and qPCR was performed according to the protocol conditions of this kit.
<Primer used>
FADS1: Mm00507605_m1
FADS2: Mm00517221_m1
ELOVL2: Mm00517086_m1
ELOVL5: Mm00506717_m1
GAPDH: Mm99999915_g1

　Figures 11A to 11D show the results of quantifying the amount of mRNA for each gene. When the Shewanella extract was given to mice, the expression of ELOVL2 was significantly promoted compared to the control group, and the expression levels of ELOVL5, FADS1 and FADS2 tended to be maintained or promoted. On the other hand, when krill oil was given to mice, it was found that the expression levels of ELOVL2, ELOVL5, FADS1 and FADS2 in the liver were significantly reduced.

[Sequence list]
SEQ ID NO: 1: Shewanella udii strain GS001 16S rDNA
caggagagta gcttgctact ttcgctgtcg agcggcggac gggtgagtaa tgcctagata
tctgcctagt cgtgggggat aacagttgga aacgactgct aataccgcat acgccctacg
ggggaaagga ggggaccttc gggcctttcg cgattagatg agtctaggtg ggattagcta
gtaggtgagg taatggctca cctaggcgac gatccctagc tgttctgaga ggatgatcag
ccacactggg actgagacac ggcccagact cctacgggag gcagcagtgg ggaatattgc
acaatgggcg aaagcctgat gcagccatgc cgcgtgtgtg aagaaggcct tcgggttgta
aagcactttc agcgaggagg aaaggttaag ggttaataac cgttagctgt gacgttactc
gcagaagaag caccggctaa cttcgtgcca gcagccgcgg taatacgagg ggtgcaagcg
ttaatcggaa ttactgggcg taaagcgtac gcaggcggtt tgttaagcca gatgtgaaag
ccccgggctc aacctgggaa ttgcatttgg aactggcaaa ctagagtctt gtagaggggg
gtagaatttc aggtgtagcg gtgaaatgcg tagagatctg aaggaatacc ggtggcgaag
gcggccccct ggacaaagac tgacgctcag gtacgaaagc gtggggagca aacaggatta
at

[Acceptance number]
(1) Identification indication: Shewanella woodyi
(2) Accession number: NITE BP-03460
(3) Date of acceptance (date of request for transfer to international deposit): April 4, 2022 (domestic deposit date: April 15, 2021)
(4) Depositary institution: National Institute of Technology and Evaluation, Patent Microorganism Depositary Center (zip code 292-0818, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan)

Claims (10)

1. glycerol-1-phosphate moiety (CH ₂ OH-CHOH-CH ₂ -OPO ₂ ^- -), ethanolamine phosphate moiety (NH ₃ ⁺ -CH ₂ -CH ₂ -OPO ₂ ^- -), and eicosapentaenoic acid moiety (CH ₃ CH ₂ (CH═CHCH ₂ ) ₅ (CH ₂ ) ₂ CO—),
   the molar ratio of ethanolamine phosphate moieties to glycerol-1-phosphate moieties is from 0.01 to 100;
   Compositions taken or administered orally.
2. The composition according to claim 1, wherein the molar ratio of said eicosapentaenoic acid moieties to the sum of said glycerol-1-phosphate moieties and ethanolamine phosphate moieties is 0.01-20.
3. Phospholipid represented by formula (I)
   

   

   (wherein R ₁ and R ₂ are selected from hydrogen atoms and acyl groups,
   at least one of R ₁ and R ₂ is an eicosapentaenoic acid moiety;
   3. The composition of claim 1 or 2, wherein X is selected from glycerol-1-phosphate moieties and ethanolamine phosphate moieties.
4. The composition according to any one of claims 1 to 3, which contains an extract of Shewanella bacteria or a mutant strain thereof.
5. The composition according to claim 4, which contains an extract of Shewanella woodyi or a mutant strain thereof.
6. The composition according to claim 5, which contains an extract of Shewanella woodyi strain GS001 (accession number NITE BP-03460) or a mutant strain thereof.
7. For at least one selected from the group consisting of increasing muscle or muscle strength, suppressing muscle or muscle strength loss, improving glucose metabolism, suppressing fat accumulation in the liver, and improving the balance of omega-3 fatty acids and omega-6 fatty acids in the body. A composition according to any one of claims 1 to 6 for use.
8. Diabetes, fatty liver, inflammation, locomotive syndrome, sarcopenia, lifestyle-related diseases and metabolic syndrome according to any one of claims 1 to 6, which is used for prevention or improvement of at least one selected from the group consisting of composition.
9. The composition according to any one of claims 1 to 6, which is used to increase at least one selected from DHA and DPA in the body.
10. The composition according to any one of claims 1 to 6, which is used for maintaining or promoting the expression of ELOVL2, ELOVL5, FADS1 or FADS2 genes in the body.
    

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