JP2023036013A - MITOCHONDRIAL FUNCTION IMPROVER, MITOCHONDRION BIOSYNTHESIS PROMOTER, AND PGC1α GENE EXPRESSION PROMOTER - Google Patents

Abstract

To provide a novel material useful as an active ingredient of an agent used to improve a condition caused by a mitochondrial dysfunction.SOLUTION: Lactic acid bacteria belonging to Fructobacillus, a culture of the lactic acid bacteria, a conditioned medium of the lactic acid bacteria, and/or extract from the lactic acid bacteria are useful as an active ingredient of a mitochondrial function improver, a mitochondrion biosynthesis promoter, and a PGC1α gene expression promoter.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to a mitochondrial function improving agent, a mitochondrial biosynthesis promoter, and a PGC1α gene expression promoter.

In today's aging society, there is growing interest in how to extend the period during which people can live independently without relying on daily or continuous medical care or nursing care, that is, how to extend their healthy life expectancy. For this reason, aging control research aimed at extending healthy life expectancy is being actively pursued.

Examples of factors known to induce aging include mitochondrial dysfunction (that is, the number and activity of mitochondria that produce energy in cells decrease with aging). Therefore, as an active ingredient for preventing anti-aging, creation of an ingredient or the like that improves the function of mitochondria is desired.

For example, Patent Document 1 discloses that a tocopheryl phosphate derivative and/or a salt thereof having a predetermined structure is an active ingredient of a mitochondrial dysfunction inhibitor, and Patent Document 2 discloses an effective Contacting cells with an amount of urolithins or their precursors is disclosed to improve mitochondrial function.

JP 2007-055923 A WO2012/088519

An object of the present invention is to provide a novel material that is useful as an active ingredient of an agent that is used for the purpose of improving conditions caused by mitochondrial dysfunction.

The present inventors have found that certain lactic acid bacteria have an action to improve mitochondrial function, an action to promote mitochondrial biosynthesis, and an action to promote PGC1α gene expression, which is a transcription coactivator involved in the regulation of mitochondrial biosynthesis gene expression. As a result, we have newly discovered that it can be an effective ingredient for improving conditions caused by mitochondrial dysfunction. In addition, although this predetermined lactic acid bacterium produces nicotinamide mononucleotide (NMN), the confirmed effect by the above action is not based on the contribution of NMN alone. It turned out to be much higher than when using In other words, it was found that the presence of components other than NMN derived from the given lactic acid bacterium contributes significantly to the mitochondrial function improving effect, mitochondrial biosynthesis promoting effect, and PGC1α gene expression promoting effect. The present invention was completed by further studies based on this finding.

That is, the present invention provides inventions in the following aspects.
Section 1. A mitochondrial function-improving agent or mitochondrial biosynthesis-enhancing agent comprising a lactic acid bacterium belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, a culture supernatant of the lactic acid bacterium, and/or an extract of the lactic acid bacterium.
Section 2.前記乳酸菌が、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＲＤ０１１７２７株（国際寄託番号ＮＩＴＥＢＰ－０２７６４）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５３株（国際寄託番号ＮＩＴＥＢＰ－０２７６５）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株（国際寄託番号ＮＩＴＥＢＰ－０２７６６）、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株からなるItem 2. The agent for improving mitochondrial function or the agent for promoting mitochondrial biosynthesis according to Item 1, which is selected from the group.
Item 3. Item 1 or 2, comprising nicotinamide adenine dinucleotide and nicotinamide mononucleotide, wherein the content ratio of said nicotinamide mononucleotide per 1 part by weight of said nicotinamide adenine dinucleotide is 0.02 parts by weight or more. The mitochondrial function-improving agent or mitochondrial biosynthesis-enhancing agent described.
Section 4. Items 1 to 3. A food, drink, cosmetic or pharmaceutical containing the mitochondrial function-improving agent or mitochondrial biosynthesis-enhancing agent according to any one of items 1 to 3.
Item 5. A PGC1α gene expression promoter comprising a lactic acid bacterium belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, a culture supernatant of the lactic acid bacterium, and/or an extract of the lactic acid bacterium.
Item 6.前記乳酸菌が、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＲＤ０１１７２７株（国際寄託番号ＮＩＴＥＢＰ－０２７６４）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５３株（国際寄託番号ＮＩＴＥＢＰ－０２７６５）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株（国際寄託番号ＮＩＴＥＢＰ－０２７６６）、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株からなるItem 6. The PGC1α gene expression promoter according to Item 5, which is selected from the group.
Item 7. Item 7. PGC1α according to Item 5 or 6, comprising nicotinamide adenine dinucleotide and nicotinamide mononucleotide, wherein the content ratio of said nicotinamide mononucleotide per 1 part by weight of said nicotinamide adenine dinucleotide is 0.02 parts by weight or more. Gene expression promoter.
Item 8. Items 5 to 7. A food, drink, cosmetic or pharmaceutical comprising the PGC1α gene expression promoter of any one of items 5 to 7.

ADVANTAGE OF THE INVENTION According to this invention, the novel raw material useful as an active ingredient of the agent used for the purpose of improving the condition resulting from mitochondrial dysfunction is provided. Specifically, according to the present invention, a lactic acid bacterium belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, and/or a culture supernatant of the lactic acid bacterium is a mitochondrial function improving agent, a mitochondrial biosynthesis accelerator, and as an active ingredient of a PGC1α expression enhancer.

FIG. 2 is a diagram showing expression levels of transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in muscle cells. FIG. 2 is a diagram showing expression levels of transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in muscle cells. FIG. 2 is a diagram showing expression levels of transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in muscle cells. FIG. 2 is a diagram showing expression levels of transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in muscle cells. FIG. 2 shows the expression level of a transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in skin cells. FIG. 2 shows the expression level of a transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in skin cells. FIG. 2 shows the expression level of a transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in skin cells. FIG. 2 shows the expression level of a transcription coactivator (PGC1α) involved in mitochondrial biosynthesis in skin cells.

The mitochondrial function-improving agent, mitochondrial biosynthesis promoter, and PGC1α gene expression promoter of the present invention contain lactic acid bacteria belonging to the genus Fructobacillus, cultures of the lactic acid bacteria, and/or culture supernatants of the lactic acid bacteria. characterized by The mitochondrial function-improving agents, mitochondrial biosynthesis promoters, and PGC1α gene expression promoters are described in detail below.

Active Ingredients The active ingredients of the mitochondrial function improving agent, mitochondrial biosynthesis promoter, and PGC1α gene expression promoter of the present invention are lactic acid bacteria belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, and a culture supernatant of the lactic acid bacterium. and/or an extract of said lactic acid bacteria.

When the active ingredient is in the form of lactic acid bacteria, the active ingredient includes bacterial components and endogenous products of the lactic acid bacterium. Lactic acid bacteria may be either viable or dead. In addition, in the case of dead bacteria, the cells may be crushed. In the case of disrupted bacteria, part of the bacterial cell components may be removed. Furthermore, the lactic acid bacteria may be in the form of powdered cells dried by freeze-drying, shelf-drying, spray-drying or the like.

When the active ingredient is a culture of lactic acid bacteria, the active ingredient includes medium components containing the products of the lactic acid bacteria as well as the lactic acid bacteria. In addition, as long as the active ingredient contains a product of the lactic acid bacterium, the medium components may be partially removed from the medium used for culturing the lactic acid bacterium. In addition, the lactic acid bacteria contained in the culture may be either viable or dead. In addition, in the case of dead bacteria, the cells may be crushed. Furthermore, in the case of disrupted bacteria, part of the bacterial cell components may be removed.

When the active ingredient is the culture supernatant of lactic acid bacteria, the culture supernatant contains medium components containing the products of the lactic acid bacteria. In addition, as long as the product of the lactic acid bacterium is contained, a part of the medium components used for culturing the lactic acid bacterium may be removed from the medium.

When the active ingredient is a lactic acid bacterium extract, the extract is a multicomponent composition obtained by subjecting the lactic acid bacterium or culture of the lactic acid bacterium to an extraction treatment. Examples of extraction solvents used for preparing the extract include water; lower alcohols such as ethanol and isopropanol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol and glycerin; and polar solvents such as mixtures thereof. From the viewpoint of more effectively improving the mitochondrial function-improving effect, the mitochondrial biosynthesis-promoting effect, or the PGC1α gene expression-promoting effect, water is preferred. Thus, in a preferred form, the extract of lactic acid bacteria is a water extract of lactic acid bacteria. Further, the temperature condition during extraction is preferably room temperature (eg, 5 to 35°C, preferably 20 to 30°C). As a specific method for preparing the extract, for example, a suspension containing the above lactic acid bacteria, a culture of lactic acid bacteria, or a pulverized product thereof in an extraction solvent is prepared, and the suspension is subjected to solid-liquid separation. A method of obtaining a liquid (extract liquid), or a method of obtaining a powder (extract powder) by drying the extract as necessary.

Lactic acid bacteria belonging to the genus Fructobacillus are microorganisms that produce nicotinamide mononucleotide (NMN). Lactic acid bacteria belonging to the genus Fructobacillus may also be microorganisms that produce nicotinamide adenine dinucleotide (NAD) in addition to NMN.

The lactic acid bacteria belonging to the genus Fructobacillus, the culture of the lactic acid bacteria, and the culture supernatant of the lactic acid bacteria, which are the active ingredients used in the present invention, contain not only NMN (or NMN and NAD) produced by the lactic acid bacteria, but also the lactic acid bacteria. It also includes other metabolites (other than NMN and NAD) produced and/or bacterial cell components. It is considered that the excellent mitochondrial function-improving effect, mitochondrial biosynthesis-promoting effect, and PGC1α gene expression-promoting effect of the present invention are brought about by the combination of these components.

Further, when the active ingredient used in the present invention contains NMN and NAD, the ratio is not particularly limited, and the ratio of NMN per 1 part by weight of NAD is, for example, 0.02 parts by weight or more, 0.035 parts by weight or more. , 0.04 parts by weight or more, or 0.05 parts by weight or more. When the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, or PGC1α gene expression-enhancing agent of the present invention is a composition for external use (preferably a composition for external use on the skin) and when used for purposes aimed at improving skin mitochondrial function, The ratio is, for example, 0.02 parts by weight or more, and from the viewpoint of further effectively improving the mitochondrial function improving effect, the mitochondrial biosynthesis promoting effect, or the PGC1α gene expression promoting effect, preferably 0.035 parts by weight. Above, more preferably 0.04 parts by weight or more, still more preferably 0.05 parts by weight or more. The upper limit of the ratio of NMN per 1 part by weight of NAD is not particularly limited, but for example, 5 parts by weight or less can be mentioned, and the effect of improving mitochondrial function, the effect of promoting mitochondrial biosynthesis, or the effect of promoting PGC1α gene expression can be improved more effectively. From the viewpoint of It is preferably 0.1 parts by weight or less, most preferably 0.07 parts by weight or less.

Alternatively, the ratio of NMN per mole of NAD is, for example, 0.04 mol or more, 0.07 mol or more, 0.08 mol or more, 0.09 mol or more, or 0.1 mol or more. When the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, or PGC1α gene expression-enhancing agent of the present invention is a composition for external use (preferably a composition for external use on the skin) and when used for purposes aimed at improving skin mitochondrial function, The ratio of NMN per mol of NAD is, for example, 0.04 mol or more, and from the viewpoint of more effectively improving the effect of improving mitochondrial function, the effect of promoting mitochondrial biosynthesis, or the effect of promoting PGC1α gene expression, it is preferably 0.04 mol or more. 07 mol or more, more preferably 0.08 mol or more, still more preferably 0.09 mol or more, and still more preferably 0.1 mol or more. The upper limit of the ratio of NMN per mol of NAD is not particularly limited, but for example, 10 mol or less can be mentioned, and from the viewpoint of more effectively improving the mitochondrial function improving effect, mitochondrial biosynthesis promoting effect, or PGC1α gene expression promoting effect. , preferably 6 mol or less, more preferably 4.6 mol or less, still more preferably 3.4 mol or less, still more preferably 2 mol or less, still more preferably 1 mol or less, particularly preferably 0.2 mol or less, most preferably Preferably 0.14 mol or less is mentioned.

Preferred examples of lactic acid bacteria belonging to the genus Fructobacillus include Fructobacillus durionis, from the viewpoint of more effectively improving the mitochondrial function-improving effect, the mitochondrial biosynthesis-promoting effect, or the PGC1α gene expression-promoting effect. Fructobacillus tropaeoil and Fructobacillus fructosus, more preferably Fructobacillus tropaeoil and Fructobacillus fructosus be done.

More preferred examples of lactic acid bacteria belonging to the genus Fructobacillus include Fructobacillus durionis strain RD011727 and Fructobacillus tropaeoil strain RD012353 from the viewpoint of more effectively improving the mitochondrial function-improving effect, mitochondrial biosynthesis-promoting effect, or PGC1α gene expression-promoting effect. 、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＮＢＲＣ１１３２３９株が挙げられ、更に好ましくは、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＲＤ０１１７２７株、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５３株、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株が挙げられる。

More preferred examples of lactic acid bacteria belonging to the genus Fructobacillus include Fructobacillus tropaeoil RD012353 strain and Fructobacillus tropaeoil RD012354 strain from the viewpoint of more effectively improving the mitochondrial function-improving effect, mitochondrial biosynthesis-promoting effect, or PGC1α gene expression-promoting effect. and Fructobacillus fructosus NBRC3516 strain, preferably Fructobacillus tropaeoil RD012353 strain and Fructobacillus fructosus NBRC3516 strain, particularly preferably Fructobacillus fructosus NBRC35 strain.

The Fructobacillus durionis strain RD011727 was deposited at the National Institute of Technology and Evaluation Patent Microorganism Depositary Center (NPMD) (2-5-8 122 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) with the deposit number dated October 28, 2020. It has been internationally deposited as NITE BP-02764.

The Fructobacillus tropaeoil strain RD012353 was deposited at the National Institute of Technology and Evaluation, Patent Microorganism Depositary Center (NPMD) (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan Room 122) with the deposit number dated October 28, 2020. It has been internationally deposited as NITE BP-02765.

The Fructobacillus tropaeoil strain RD012354 was deposited at the National Institute of Technology and Evaluation Patent Microorganisms Depositary Center (NPMD) (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan Room 122) with a deposit number dated October 28, 2020. It has been internationally deposited as NITE BP-02766.

One of these fructobacillus lactic acid bacteria may be used alone, or two or more of them may be used in combination.

A culture or culture supernatant of the fructobacillus lactic acid bacterium can be obtained by culturing under conditions in which the above fructobacillus lactic acid bacterium produces nicotinamide mononucleotide. The medium and culture conditions used for culturing fructobacillus lactic acid bacteria can be appropriately selected.

Media include those used for expansion culture (pre-culture medium) and those used for production culture (main culture medium). The main culture medium can be prepared based on the medium used as the pre-culture medium and further containing additives. Regarding the properties of the medium, it is preferably a liquid medium, but may be an agar medium. The medium generally contains a nitrogen source, minerals, etc., in addition to the carbon source.

Carbon sources include carbohydrates and carbohydrate materials. Carbohydrates include sugars (monosaccharides, disaccharides, oligosaccharides), polysaccharides, and sugar alcohols. Carbohydrates include lactose, sucrose, glucose, starch, xylitol, dextrose and the like. The carbohydrate material may be any organic composition containing carbohydrates, such as milk and its processed products (skimmed milk powder, whey, milk powder, condensed milk, etc.), soymilk and its processed products (soymilk hydrolyzate, etc.), Foods such as cereals, fruits and vegetables can be mentioned. Milk includes that from any mammal such as cows, goats, sheep, buffaloes, camels, llamas, donkeys, yaks, horses, reindeer, and the like. The sugar may be isolated or contained in sugar material. For example, fructose (sugar) may be in the form contained in fruits (sugar material). One of these carbon sources may be used alone, or two or more of them may be used in combination. Among these carbon sources, glucose is preferred.

The concentration of the carbon source in the medium is not particularly limited, and may be appropriately set according to the type of medium and culture method. More preferably 1.5 to 2.5 w/w%.

Any inorganic or organic nitrogen source can be used as the nitrogen source. For example, proteins such as yeast extract (brewer's yeast, etc.), meat extract, casein; protein hydrolysates such as peptone (protease peptone, etc.); peptides such as peptides; nitrogen salts and the like. One of these nitrogen sources may be used alone, or two or more of them may be used in combination.

The concentration of the nitrogen source in the medium is not particularly limited, and may be appropriately set according to the type of medium, culture method, etc. In the case of protein, for example, 0.3 to 4 w/w%, preferably 0.5. ~3 w/w%, more preferably 1-2 w/w%; 0.5 to 1.5 w/w%; in the case of nitrogen-containing salts, for example 0.03 to 1.5 w/w%, preferably 0.05 to 1 w/w%, more preferably 0.1 to 0.5 w/w% is mentioned.

Examples of minerals include manganese (e.g., manganese salts such as manganese sulfate), zinc, iron, sodium (e.g., sodium salts such as sodium acetate), potassium (e.g., dipotassium hydrogen sulfate, dipotassium hydrogen phosphate, etc.). potassium salt), magnesium (e.g. magnesium salt such as magnesium sulfate), calcium, phosphorus (e.g. phosphate such as dipotassium hydrogen phosphate), sulfur (e.g. sulfuric acid such as manganese sulfate, potassium hydrogen sulfate, magnesium sulfate) salts), trace elements, and the like. These minerals may be used individually by 1 type, and may be used in combination of multiple types. Among these minerals, manganese, sodium, magnesium and potassium are preferred.

The concentration of minerals in the medium is not particularly limited, and may be appropriately set according to the type of medium and culture method. .003 to 0.008 w/w%; for sodium salts, for example 0.05 to 1.5 w/w%, preferably 0.1 to 1 w/w%; for magnesium salts, for example 0.001-0.02 w/w%, preferably 0.005-0.015 w/w%; in the case of potassium salts, for example 0.05-1 w/w%, preferably 0.1-0. 5 w/w%; for phosphates, for example 0.05 to 1 w/w%, preferably 0.1 to 0.5 w/w%; for sulfates, 0.001 to 0 0.04 w/w%, preferably 0.005-0.02 w/w%.

In addition to the above components, the medium contains vitamins (Vitamin B group, etc.), surfactants (nonionic surfactants (Tween, etc.), anionic surfactants (SDS, etc.), etc.), antibacterial agents (Triclosan, etc.). , antibiotics (monesin, etc.). These other components may be used individually by 1 type, and may be used in combination of multiple types. Among these other ingredients, preferably surfactants, more preferably nonionic surfactants are included.

The concentration of other components in the medium is not particularly limited, and may be appropriately set according to the type of other components, the type of medium, the culture method, etc. However, when a surfactant is included, the concentration of the surfactant As, for example, 0.01 to 0.5 w/w%, preferably 0.05 to 0.3 w/w%.

Culture conditions are not particularly limited as long as they are conditions under which the fructobacillus lactic acid bacteria can grow.

The culture temperature may be the optimum temperature for the fructobacillus lactic acid bacteria to be cultured, for example, 26 to 40° C., preferably 27 to 38° C., more preferably 28 to 36° C., still more preferably 29 to 34° C. °C. The culture time may be appropriately set according to the type of fructobacillus lactic acid bacteria to be actually cultured, for example, 4 to 48 hours, preferably 8 to 36 hours, more preferably 12 to 24 hours.

Regarding the operation during culture, it does not matter whether the culture solution needs to be stirred during culture. In addition, as an example of a specific culture procedure, the above culture is performed as a production culture (main culture) for a certain period of time, and before that, a small amount (eg, 1/6 to 1/4 of the main culture medium in volume ratio) An expansion culture (pre-culture) can be performed in the medium. The culture conditions for the pre-culture may be appropriately set according to the type of fructobacillus lactic acid bacteria, and the above conditions can be adopted. In the main culture, the culture obtained in the preculture can be inoculated into the main culture medium so that the OD660 is, for example, 0.01 to 0.04, preferably 0.01 to 0.03. can.

The culture or culture supernatant of these fructobacillus lactic acid bacteria may be prepared using one of the above fructobacillus lactic acid bacteria alone, or may be prepared using a combination of a plurality of types. However, it may be a mixture of one prepared by using one type alone and one prepared by using another type alone.

As the lactic acid bacteria belonging to the genus Fructobacillus used in the present invention, lactic acid bacteria after culture may be used as they are. may be used.

The lactic acid bacteria belonging to the genus Fructobacillus obtained by culturing may be used directly as an active ingredient of a mitochondrial function improving agent, or may be filtered using filter paper, centrifuged, decanted, screw pressed, roller pressed, rotary drum. Screen, belt screen, vibrating screen, multi-plate vibrating filter, vacuum dehydration, pressure dehydration, belt press, centrifugal concentration dehydration, multi-disk dehydration, etc. are used for solid-liquid separation. The microbial cells obtained by the reaction may be used as the active ingredient of the mitochondrial function-improving agent.

When the cultured lactic acid bacteria belonging to the genus Fructobacillus are used for the resting cell reaction, the nicotinamide mononucleotide content in the lactic acid bacteria can be increased.

The liquid used for the resting cell reaction is not particularly limited as long as it allows the lactic acid bacteria belonging to the genus Fructobacillus to react efficiently. For example, water, buffer solutions, organic solvents, and the like can be used.

The pH of the liquid used for the resting cell reaction is, for example, 4.0 to 10.0, preferably 5.0 to 9.0, more preferably 5.5 to 7.5.

When the liquid used for the resting cell reaction is a buffer, examples of the buffer include acetate buffer, phosphate buffer, borate buffer, carbonate buffer, citrate buffer, Tris buffer, and HEPES buffer. , MES buffer and the like.

More specific examples of buffers include KHC ₈ H ₄ O ₄ -NaOH (pH 4.0), CH ₃ COOH-CH ₃ COONa (pH 4.0), MES-NaOH (pH 5.0), CH ₃ COOH -CH ₃ COONa (pH 5.0), KH ₂ PO ₄ -K ₂ HPO 4 (pH 6.0), MES-NaOH (pH 6.0), KH ₂ PO ₄ -K ₂ HPO ₄ (pH 7.0), PIPES- NaOH (pH 7.0), HEPES-NaOH (pH 8.0), H ₃ BO ₄ -NaOH (pH 8.0), CHES-NaOH (pH 9.0), H ₃ BO ₄ -NaOH (pH 9.0), H ₂ CO ₃ —NaHCO ₃ (pH 10.0), CHES-NaOH (pH 10.0), etc., preferably CH ₃ COOH—CH ₃ COONa, KH ₂ PO ₄ —K ₂ HPO ₄ , H ₃ BO ₄ — NaOH, more preferably KH ₂ PO ₄ -K ₂ HPO ₄ .

When the liquid used for resting cell reaction is an organic solvent, examples of the organic solvent include aromatic compounds such as benzene and benzonitrile, ketones such as acetone, acetylacetone and methyl ethyl ketone, ethyl acetate, butyl acetate, ethyl butyrate, Fatty acid esters such as ethyl formate, ethers such as diethyl ether, dipropyl ether, dibutyl ether, tetrahydrofuran and 1,4-dioxane, halogenated hydrocarbons such as dichloromethane, chloroform and dichloroethane, 1,2-propanediol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol, 1,2-hexanediol, 1,6-hexanediol, 1,2-pentanediol, 1 ,5-pentanediol, 2-methyl-2,4-pentanediol, 3-methyl-1,5-pentanediol and other diols, alcohols having a linear or branched alkyl group having 1 to 7 carbon atoms, cyclohexanol, alcohols such as 3-methoxy-3-methyl-1-butanol and 3-methoxy-1-butanol;

Among the above-mentioned liquids, the liquid used for the resting cell reaction is preferably water or a buffer solution.

The reaction temperature for resting cell reaction is, for example, 21 to 37°C, preferably 24 to 28°C. The reaction time is, for example, 0.1 to 24 hours, preferably 6 to 12 hours.

The resting cell reaction can be carried out by suspending lactic acid bacteria belonging to the genus Fructobacillus in the above liquid, and allowing the suspension to stand, stirring, or shaking.

In the resting cell reaction, the pH of the culture solution containing lactic acid bacteria belonging to the genus Fructobacillus is 4.0 to 10.0, preferably 5.0 to 9.0, more preferably 5.0 to 9.0, without using the above liquid. may be adjusted in the range of 5.5 to 7.5.

Other Ingredients The mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, and PGC1α gene expression-enhancing agent of the present invention may optionally contain other pharmacological ingredients in addition to the above active ingredients. Examples of such pharmacological components include anti-inflammatory agents, antioxidants, bactericidal agents, cooling agents, vitamins, mucopolysaccharides and the like.

In addition, the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, and PGC1α gene expression-enhancing agent of the present invention may contain bases and additives as necessary in order to obtain desired formulation forms. . Such bases and additives are not particularly limited as long as they are pharmaceutically acceptable. Examples include water, lower alcohols (ethanol, isopropanol, etc.), polyhydric alcohols (glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, etc.); oily bases such as naturally derived oils, mineral oils, ester oils, fatty acid alkyl esters, fatty acids, fatty acid esters, higher alcohols; surfactants; , preservatives, flavoring agents, coloring agents, thickeners, pH adjusters, wetting agents, stabilizers, antioxidants, UV absorbers, chelating agents, adhesives, buffering agents, solubilizers, solubilizers , preservatives and other additives.

Properties and Forms The properties of the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, and PGC1α gene expression-enhancing agent of the present invention are not particularly limited. , paste) or the like. In addition, the mitochondrial function-improving agent, mitochondrial biosynthesis promoter, and PGC1α gene expression promoter of the present invention may be non-emulsified preparations such as aqueous preparations and oil-based preparations. It may be an emulsified formulation such as a type emulsified formulation.

In addition, the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, and PGC1α gene expression-enhancing agent of the present invention can be used in the form of a composition for internal use or a composition for external use (preferably a composition for external use on the skin).

Specifically, the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent, and PGC1α gene expression-enhancing agent of the present invention can be used in the form of foods, drinks, cosmetics, and pharmaceuticals (oral or topical medicines).

Food and drink are not particularly limited, but for example, fermented milk (drink yogurt, etc.), lactic acid bacteria drinks, milk drinks (coffee milk, fruit milk, etc.), tea drinks (green tea, black tea, oolong tea, etc.), fruit and vegetable drinks (beverages containing fruit juices such as oranges, apples, grapes, etc., vegetable juices such as tomatoes, carrots, etc.), alcoholic beverages (beer, low-malt beer, wine, etc.), carbonated beverages, soft drinks, water-based beverages; and Examples include foods such as fermented milk (set type yogurt, soft yogurt, etc.), confectionery, instant foods, and processed foods such as seasonings.

Moreover, functional food is also mentioned as food and drink. Functional foods refer to foods that have a certain level of functionality for living organisms. Foods with sex indications, foods for special uses, nutritional supplements, health supplements, supplements (for example, tablets, coated tablets, sugar-coated tablets, capsules, liquid preparations, etc.), beauty foods (for example, diet foods), etc. . Furthermore, the functional food may be food for special uses such as food for sick people, powdered milk for pregnant and breastfeeding women, powdered milk for infants, food for the elderly, food for nursing care, and the like.

Cosmetics include lotions, milky lotions, creams, essences, gels, packs, sheet masks, lip balms, and other basic cosmetics; facial cleansers, makeup removers (including cleansing agents), exfoliants, body shampoos, and other skin cleansers. Cosmetics: Body care cosmetics such as sunscreens, body gels, body massage agents, antiperspirants, deodorants, hair removers, and bath salts; foundations, powders, lipsticks, blushers, eyeshadows, eyeliners, mascara , makeup cosmetics such as eyebrows; nail cosmetics such as manicures and nail removers; hair cosmetics such as hair styling agents, shampoos, conditioners, rinses, and hair restorers; liquid toothpaste, toothpaste, mouthwash, mouth spray Oral cosmetics such as

Pharmaceuticals include internal medicines and external medicines (including quasi-drugs). Dosage forms of oral medicines include tablets, coated tablets, sugar-coated tablets, capsules, liquids, etc. Dosage forms of external medicines include liquids (including lotions, sprays, aerosols, and emulsions) and foams. , ointments, creams, gels, patches and the like.

Preferred subjects for application of foods, cosmetics, or pharmaceuticals containing mitochondrial function improving agents, mitochondrial biosynthesis promoters, or PGC1α gene expression promoters include mammals, for example, primates such as humans, chimpanzees, and gorillas. pets or farm animals such as dogs, cats, rabbits, cows, horses, pigs, goats, sheep and donkeys; laboratory animals such as mice, rats, hamsters and monkeys.

Uses The mitochondrial function-improving agent of the present invention is used for the purpose of improving mitochondrial function (that is, for the purpose of improving conditions caused by decreased mitochondrial function). The target mitochondria are not particularly limited, but preferably include skin mitochondria and muscle mitochondria, and more preferably include skin mitochondria. Specific uses aimed at improving mitochondrial function include those aimed at improving skin mitochondrial function, such as suppressing or improving skin wrinkle formation; improving muscle mitochondrial function Examples of intended purposes include uses such as improving exercise capacity and suppressing muscle weakness; other examples include uses such as improving metabolic function, extending life span, preventing aging, recovering from abnormal nervous system, and improving cognitive function. is mentioned.

The mitochondrial biosynthesis-promoting agent of the present invention is used for purposes aimed at promoting mitochondrial biosynthesis (and for purposes aimed at improving conditions caused by decreased mitochondrial function). The target mitochondria are not particularly limited, but preferably include skin mitochondria and muscle mitochondria, and more preferably include skin mitochondria. Specific uses aimed at promoting mitochondrial biosynthesis include those aimed at promoting skin mitochondrial biosynthesis, for example, uses for suppressing or improving skin wrinkle formation; muscle mitochondrial biosynthesis; For the purpose of promoting, for example, the use of improving exercise capacity, suppressing muscle weakness, etc.; and other applications.

The PGC1α gene expression promoter of the present invention is used for the purpose of promoting PGC1α gene expression. Specific uses aimed at promoting PGC1α gene expression are not particularly limited, but include uses such as promoting mitochondrial biosynthesis (and thus improving conditions caused by decreased mitochondrial function) and promoting myokine production. . The target tissue for PGC1α gene expression is not particularly limited, but skin and muscle are preferred, and skin is more preferred. More specific uses aimed at promoting PGC1α gene expression include uses aimed at promoting PGC1α gene expression in the skin, such as suppressing or improving skin wrinkle formation; Uses for promoting the expression of the PGC1α gene include, for example, amelioration of decreased exercise capacity and suppression of muscle weakness.

Dosage When the agent for improving mitochondrial function, the agent for promoting mitochondrial biosynthesis and the agent for promoting PGC1α gene expression of the present invention is used internally, the amount of the active ingredient is, for example, 0.01 to 200 mg/kg/day, preferably 0.1 to 200 mg/kg/day. It can be used internally in an amount of 20 mg/kg/day 1 to 5 times a day.

When the mitochondrial function-improving agent, mitochondrial biosynthesis-enhancing agent and PGC1α gene expression-enhancing agent of the present invention are externally applied, the amount of the active ingredient is, for example, 0.0001 to 0.2 mg/cm ² per application amount, It can be applied externally 1 to 5 times a day, preferably in an amount of 0.001 to 0.02 mg/cm ² .

EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to these.

Test example 1
(1) Preparation of Fructobacillus lactic acid bacterium powder (Sample A) Fructobacillus fructosus NBRC3516 strain was inoculated into 30 ml of Difco MRS medium (preculture medium), and expanded culture was allowed to stand at 30°C for 24 hours. bottom. The resulting culture solution was inoculated into 1 L of MRS medium (main culture medium) so that OD660 was 0.02, and cultured with stirring at pH 6.0 to 7.0 at 30° C. for 12 hours.
The resulting culture solution was subjected to centrifugation to recover the cells. The recovered cells were washed with 1 L of 0.85 w/w % KCl aqueous solution. The washed cells were subjected to centrifugation again to recover the cells. After the collected cells were sterilized, they were freeze-dried to obtain lactic acid bacteria powder (Sample A). Analysis was performed under the HPLC conditions described later, and the amounts of nicotinamide mononucleotide and nicotinamide adenine dinucleotide production in the collected cells were measured. As a result of measurement, the weight ratio of NMN to 1 weight part of NAD in sample A was 0.055 weight part. Sample A was used in Examples 1 and 2.

(2) Preparation of Fructobacillus lactic acid bacteria powder (Sample B) Fructobacillus fructosus NBRC3516 strain was inoculated into 30 ml of Difco MRS medium (pre-culture medium), and expanded culture was allowed to stand at 30°C for 24 hours. bottom. The resulting culture solution was inoculated into 1 L of MRS medium (main culture medium) so that OD660 was 0.02, and cultured with stirring at pH 6.0 to 7.0 at 30° C. for 12 hours.
The resulting culture solution was subjected to centrifugation to recover the cells. The recovered cells were washed with 1 L of 0.85 w/w % KCl aqueous solution. The washed cells were subjected to centrifugation again to recover the cells.
The collected cells were suspended in 200 ml of deionized water and stirred at pH 6.0 to 7.0 at 25° C. for 8 hours to carry out resting cell reaction. After completion of resting cell reaction, cells were collected by centrifugation. The recovered cells were washed with 1 L of 0.85 w/w % KCl aqueous solution. The washed cells were subjected to centrifugation again to recover the cells. After the collected cells were sterilized, they were freeze-dried to obtain lactic acid bacteria powder (sample B). It was analyzed under the HPLC conditions described later, and the production amounts of nicotinamide mononucleotide and nicotinamide adenine dinucleotide in the collected cells were measured. As a result of the measurement, the weight ratio of NMN per part by weight of NAD in sample B was 0.55 parts by weight Sample B was used in Examples 3 and 4.

(MRS medium composition)
2 w/w% glucose 1 w/w% protease peptone 1 w/w% beef extract 0.5 w/w% yeast extract 0.2 w/w% ammonium citrate 0.1 w/w% Tween80
0.5 w/w% sodium acetate 0.01 w/w% magnesium sulfate 0.005 w/w% manganese sulfate 0.2 w/w% dipotassium hydrogen phosphate

(HPLC analysis conditions) NMN measurement method Column: DaisoPak SP-100-5-ODS-P (4.6 × 150 mm) × 2 column temperature: 25 ° C.
Eluent: 75 mM ammonium phosphate aqueous solution (pH 6.0)
Flow rate: 0.6ml/min
Detector: UV detector (260 nm)
Detection time: 14.5 minutes

(HPLC analysis conditions) Method for measuring NAD Column: DAISOPAK SP-100-5-ODS-P (4.6 x 250 mm) x 2 Column temperature: 25°C
Eluent: 75 mM ammonium phosphate aqueous solution (pH 6.0): methanol = 95: 5 (w: w)
Flow rate: 0.7mL/min
Detector: UV detector (260 nm)
Detection time: 33 minutes

(3) Mitochondrial function improvement test and results C2C12 cells were seeded on a 96-WELL μClear fluorescence black plate (Greiner-Bio One, Tokyo, Japan) at 1.0 × 10 ⁴ cells/well, 37 ° C., 5% Cultured for 24 hours in the presence of _CO2 . After that, the components shown in Table 1 were added to the water and cultured for 24 hours. After that, the cells were stained with Hoechst33342 and MitoTracker Green FM, and the number of mitochondria was analyzed with IN CELL Analyzer2200. Assuming that the number of mitochondria in Comparative Example 1 (control to which only water was added) was 100, the relative number of mitochondria in each example and each comparative example was derived. When the mitochondrial relative number exceeds 100, it can be evaluated that mitochondrial biosynthesis is promoted, that is, mitochondrial function is improved. Table 1 shows the results.

Furthermore, for Comparative Examples 1 and 4 and Examples 3 and 4, the area of mitochondria stained with MitoTracker Green FM was analyzed using an IN CELL Analyzer 2200. Assuming that the mitochondrial area in Comparative Example 1 (control to which only water was added) was 100, the relative area of mitochondria in each Example and Comparative Example was derived. When the mitochondrial relative area exceeds 100, it can also be evaluated that mitochondrial biosynthesis is promoted, that is, mitochondrial function is improved. Table 2 shows the results.

As shown in Table 1, NAD and NMN as reagents could not improve mitochondrial function at the amounts used in Comparative Examples 2 and 4, but as shown in Examples 1 to 4, Fructobacillus It was confirmed that the lactic acid bacteria powder of the genus significantly improved mitochondrial function even though the contents of NAD and NMN were much lower than those of Comparative Example 2 or Comparative Example 3. The results shown in Table 2 also reflect the trends shown in Table 1 well. In addition, as shown in Comparative Examples 3 and 5 in Table 1, NAD and NMN as reagents, even if the amount was increased to 10 times the amount used in Comparative Examples 2 and 4, substantial mitochondrial function was observed. No improvement effect was obtained. From these facts, it can be said that the mitochondrial function-improving effect of the fructobacillus lactic acid bacteria powder is astonishing.

Test example 2
(1) Preparation of fructobacillus lactic acid bacterium extract (sample a) 20 g of the dried lactic acid bacterium (sample A) prepared in Test Example 1 was finely ground in a mortar and suspended in 1 L of sterilized water to prepare a suspension. The suspension was stirred at room temperature (about 25° C.) for 1 hour, centrifuged (12,000 rpm×10 min), and the supernatant was filtered through a 0.2 μm filter to obtain a lactic acid bacterium extract (sample a). The nicotinamide mononucleotide (NMN) and nicotinamide adenine dinucleotide (NAD) production amounts in the obtained sample a were measured under the HPLC conditions described above. As a result of measurement, the weight ratio of NMN to 1 part by weight of NAD in sample a was 0.0527 parts by weight. Sample a was used in Examples 5 and 9.

(2) Preparation of other fructobacillus lactic acid bacteria extracts (samples c, d, and e) A powder of each lactic acid bacterium was prepared in the same manner as in "(1) Preparation of powder of fructobacillus lactic acid bacterium (sample A)". Using the obtained powder of each lactic acid bacterium, an extract was prepared in the same manner as in "(1) Preparation of extract of fructobacillus lactic acid bacteria (sample a)" in this test example. Specifically, the extract (sample c) was prepared from the powder of Fructobacillus tropaeoil RD012353 strain, the extract (sample d) from the powder of Fructobacillus tropaeoil RD012354 strain, and the extract (sample d) from the powder of Fructobacillus durionis RD011727 strain. Sample e) was obtained. Using the methods of "(HPLC analysis conditions) NMN measurement method" and "(HPLC analysis conditions) NAD measurement method" in Test Example 1, nicotinamide mononucleotide (NMN) and nicotinamide in samples c, d, and e Adenine dinucleotide (NAD) production was measured. As a result of measurement, the weight ratio of NMN to 1 part by weight of NAD in sample c was 0.0202 parts by weight. The weight ratio of NMN per part by weight of NAD in sample d was 0.0250 parts by weight. The weight ratio of NMN to 1 weight part of NAD in sample e was 0.0421 weight part. Sample c was used for Examples 6 and 10, sample d was used for Examples 7 and 11, and sample e was used for Examples 8 and 12.

(3) Measurement and results of expression level of transcription coactivator PGC1α involved in gene expression of mitochondrial biogenesis in mouse myoblasts (Examples 5-8)
Mouse myoblasts C2C12 (RIKEN Cell Bank, RCB0987) were seeded in a 6-well plate and placed in DMEM (Fujifilm Wako Pure Chemical) medium containing 10 wt% fetal bovine serum at 37°C under 5 vol% _CO2 conditions. cultured in When subconfluent, the medium was replaced with DMEM medium containing 2 vol% horse serum and cultured for 6 days.

Each of samples a, c to d was replaced with DMEM medium containing 2% by volume horse serum added to a final concentration of 1% by volume, and cultured for 24 hours at 37° C. and 5% by volume CO ₂ . As controls, the same operation was performed except that samples a and cd were not added. Total RNA was extracted from the resulting culture.

Total RNA was extracted using ISOGEN (Nippon Gene) after washing the C2C12 cells with PBS. Total RNA was quantified using the Qubit RNA HS Assay Kit (Invitrogen). The extracted RNA was reverse transcribed using ReverTra qPCR RT Master Mix (Toyobo) to synthesize single-stranded cDNA. Using this cDNA as a template, real-time PCR was performed with THUNDERBIRD NextSYBR qPCR Mix (Toyobo) using the fluorescent dye CyberGreen. In the PCR reaction, after initial denaturation at 95°C for 30 seconds, amplification reaction at 95°C for 5 seconds and 60°C for 30 seconds was repeated 40 times. The gene expression level was calculated as a relative value from the number of cycles until a constant amount of PCR product was obtained. Real-time PCR was performed on the GAPDH gene as an internal standard in addition to the PGC1α gene, and evaluation was performed by correcting the relative expression level of PGC1α with the relative expression level of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). .

As primers for amplifying PGC1α, sense: 5′-GAATCAAGCCACTACAGACACCG-3′ (SEQ ID NO: 1), antisense: 5′-CATCCCTCTTGAGCCTTTCGTG-3′ (SEQ ID NO: 2), as primers for amplifying GAPDH, 5′-CATCACTGCCCACCCAGAAGACTG-3′ (SEQ ID NO: 3), antisense: 5′-ATGCCAGTGAGCTTCCGTTCAG-3′ (SEQ ID NO: 4) was used.

Gene expression levels were measured by real-time PCR. The PGC1α gene expression level was determined as a ratio to the internal standard GAPDH (glyceraldehyde-3-phosphate dehydrogenase) gene expression level. When the PGC1α gene expression level in the control was set to 1, the relative amount of PGC1α gene expression due to the addition of each lactic acid bacteria extract was calculated. The results are shown in FIGS. 1-4.

As shown in FIGS. 1 to 4, the fructobacillus lactic acid bacterium extract markedly increased the expression level of the transcription coactivator PGC1α gene, which is involved in gene expression of mitochondrial biosynthesis, in mouse myoblasts compared to no lactic acid bacterium extract. It was confirmed that it was increased.

(4) Measurement and results of expression level of transcription coactivator PGC1α involved in gene expression of mitochondrial biogenesis in human epidermal keratinocytes (Examples 9 to 12)
Human epidermal keratinocytes PSVK1 (JCRB Cell Bank, JCRB1093 cell founder: Yasumoto, S.) were seeded in a 6-well plate at 5×10 ⁵ cells/well, and placed under the conditions of 37° C. and 5 vol% CO _{2 .} was cultured overnight. The medium was a KMG-Gold basal medium (Lonza) medium supplemented with an additive (KGM-Gold Single Quots (Lonza)) and cultured for 24 hours at 37° C. under 5 vol % CO ₂ conditions.

After culturing, the medium was changed to KMG-Gold basal medium and cultured for 24 hours at 37° C. and 5 vol % CO ₂ . Thereafter, each of samples a, c to d was replaced with a KMG-Gold basal medium added to a final concentration of 1% by weight, and cultured for 3 hours at 37° C. and 5 vol % CO ₂ . As controls, the same operation was performed except that samples a and cd were not added. Total RNA was extracted from the resulting culture.

For extraction of total RNA, PSVK1 cells were washed with PBS and then extracted using ISOGEN (Nippon Gene). Total RNA was quantified using the Qubit RNA HS Assay Kit (Invitrogen). The extracted RNA was reverse transcribed using ReverTra qPCR RT Master Mix (Toyobo) to synthesize single-stranded cDNA. Using this cDNA as a template, real-time PCR was performed with THUNDERBIRD NextSYBR qPCR Mix (Toyobo) using the fluorescent dye CyberGreen. In the PCR reaction, after initial denaturation at 95°C for 30 seconds, amplification reaction at 95°C for 5 seconds and 60°C for 30 seconds was repeated 40 times. The gene expression level was calculated as a relative value from the number of cycles until a constant amount of PCR product was obtained. Real-time PCR was performed on the GAPDH gene as an internal standard in addition to the PGC1α gene, and evaluation was performed by correcting the relative expression level of PGC1α with the relative expression level of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). .

As primers for amplifying PGC1α, sense: 5′-CCAAAGGATGCGCTCTCGTTCA-3′ (SEQ ID NO: 5), antisense: 5′-CGGTGTCTGTGTAGTGGCTTGACT-3′ (SEQ ID NO: 6) as primers for amplifying GAPDH, 5′-GTCTCCTCTGACTTCAACAGCG-3′ (SEQ ID NO: 7), antisense: 5′-ACCACCCTGTTGCTGTAGCCAA-3′ (SEQ ID NO: 8) was used.

Gene expression levels were measured by real-time PCR. The PGC1α gene expression level was determined as a ratio to the internal standard GAPDH (glyceraldehyde-3-phosphate dehydrogenase) gene expression level. When the PGC1α gene expression level in the control was set to 1, the relative amount of PGC1α gene expression due to the addition of the lactic acid bacteria extract was calculated. The results are shown in FIGS. 5-8.

As shown in FIGS. 5 to 8, the fructobacillus lactic acid bacterium extract significantly increases the expression level of the transcription coactivator PGC1α gene, which is involved in gene expression of mitochondrial biosynthesis in human epidermal keratinocytes, compared to the case where no lactic acid bacterium extract is added. It was confirmed that it was increased.

Claims (8)

A mitochondrial function-improving agent or mitochondrial biosynthesis-enhancing agent comprising a lactic acid bacterium belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, a culture supernatant of the lactic acid bacterium, and/or an extract of the lactic acid bacterium. 前記乳酸菌が、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＲＤ０１１７２７株（国際寄託番号ＮＩＴＥＢＰ－０２７６４）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５３株（国際寄託番号ＮＩＴＥＢＰ－０２７６５）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株（国際寄託番号ＮＩＴＥＢＰ－０２７６６）、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株からなるThe mitochondrial function-improving agent or mitochondrial biosynthesis-enhancing agent according to claim 1, which is selected from the group. 2. Mitochondrial function according to claim 1, comprising nicotinamide adenine dinucleotide and nicotinamide mononucleotide, wherein the content ratio of said nicotinamide mononucleotide per 1 part by weight of said nicotinamide adenine dinucleotide is 0.02 parts by weight or more. ameliorator or mitochondrial biosynthesis promoter. Food, beverages, cosmetics or pharmaceuticals comprising the mitochondrial function-improving agent or mitochondrial biosynthesis accelerator according to claim 1. A PGC1α gene expression promoter comprising a lactic acid bacterium belonging to the genus Fructobacillus, a culture of the lactic acid bacterium, a culture supernatant of the lactic acid bacterium, and/or an extract of the lactic acid bacterium. 前記乳酸菌が、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｄｕｒｉｏｎｉｓ ＲＤ０１１７２７株（国際寄託番号ＮＩＴＥＢＰ－０２７６４）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５３株（国際寄託番号ＮＩＴＥＢＰ－０２７６５）、Ｆｒｕｃｔｏｂａｃｉｌｌｕｓ ｔｒｏｐａｅｏｉｌ ＲＤ０１２３５４株（国際寄託番号ＮＩＴＥＢＰ－０２７６６）、及びＦｒｕｃｔｏｂａｃｉｌｌｕｓ ｆｒｕｃｔｏｓｕｓ ＮＢＲＣ３５１６株からなるThe PGC1α gene expression promoter according to claim 5, which is selected from the group. The PGC1α gene according to claim 5, comprising nicotinamide adenine dinucleotide and nicotinamide mononucleotide, wherein the content ratio of said nicotinamide mononucleotide per 1 part by weight of said nicotinamide adenine dinucleotide is 0.02 parts by weight or more. Expression promoter. Food, beverages, cosmetics or pharmaceuticals comprising the PGC1α gene expression promoter according to claim 5 .

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