Composition for enhancing the physiological activity effect of lactic acid bacteria

Abstract

The present invention relates to a composition for enhancing the effect of lactic acid bacteria, and more specifically, by containing at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine and valine, the physiological activity of lactic acid bacteria, such as the growth-promoting effect or intestinal damage-inhibiting effect of lactic acid bacteria, can be enhanced.

Description

Technical Field

[0001] The present invention relates to a composition for enhancing the physiological activity effect of lactic acid bacteria.

Background Art

[0002] Lactic acid bacteria are bacteria that produce lactic acid using sugar as an energy source, and are present in the digestive organs and oral cavity of humans and mammals, and are widely distributed in nature such as various fermented foods. Lactic acid bacteria are one of the microorganisms that humans have used widely for the longest time, and are microorganisms that do not produce substances harmful to the intestines of humans and animals and have the function of preventing putrefaction in the intestine.

[0003] Lactic acid bacteria are typical probiotics, and not only have an antibacterial effect, but also regulate the host's intestinal microflora, and show beneficial effects on humans in various aspects such as suppression of various intestinal diseases and enhancement of immunity. Therefore, there is an increasing interest in developing them as various food materials. Such lactic acid bacteria are widely used in human life, ranging from various fermented foods centered on fermented dairy products to sauces, beverages, pharmaceuticals, and feed additives for livestock.

[0004] On the other hand, lactic acid bacteria products use prebiotic additives such as vitamins and dietary fiber together for the purpose of increasing the effects of lactic acid bacteria. Conventional or current lactic acid bacteria-related products are simply combinations of a plurality of components, so problems may occur such as not being able to show a great effect, or having an adverse effect on the metabolism of the taker when taken in large amounts, and there is a lack of research on how additives affect the physiological activity of lactic acid bacteria.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The present invention provides a composition for enhancing the effects of lactic acid bacteria.

[0006] The present invention provides a pharmaceutical composition for promoting growth or inhibiting intestinal injury.

[0007] The present invention provides a food composition for promoting growth or inhibiting intestinal injury.

Means for Solving the Problems

[0008] 1. A composition for enhancing the effect of lactic acid bacteria containing at least two branched-chain amino acids (BCAAs) selected from the group consisting of leucine, isoleucine, and valine.

[0009] 2. The composition for enhancing the effect of lactic acid bacteria according to item 1 above, which contains the leucine, isoleucine, and valine.

[0010] 3. The composition for enhancing the effect of lactic acid bacteria according to item 1 above, wherein the effect of the lactic acid bacteria is to promote the growth of an individual or inhibit intestinal injury.

[0011] 4. The composition for enhancing the effect of lactic acid bacteria according to item 1 above, wherein the effect of the lactic acid bacteria is to increase at least one of the body weight or bone density of a nutritionally deficient individual or inhibit intestinal leakage of a nutritionally deficient individual.

[0012] 5. The composition for enhancing the effect of lactic acid bacteria according to item 1 above, wherein the lactic acid bacteria is at least one selected from the group consisting of Lactobacillus spp., Lactococcus spp., Enterococcus spp., Streptococcus spp., and Bififobacterium spp.

[0013] 6. In Item 5 above, the Lactobacillus genus is at least one selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius, a composition for enhancing the effects of lactic acid bacteria.

[0014] 7. A pharmaceutical composition for promoting growth or inhibiting intestinal injury, comprising the composition according to any one of Items 1 to 6 above and lactic acid bacteria.

[0015] 8. In item 7 above, the lactic acid bacterium is at least one selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius, and is a pharmaceutical composition.

[0016] 9. A pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of growth disorder, growth retardation, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal leakage syndrome, which comprises the composition according to any one of items 1 to 6 above and a lactic acid bacterium.

[0017] 10. A food composition for promoting growth or inhibiting intestinal injury, which comprises the composition according to any one of items 1 to 6 above and a lactic acid bacterium.

Advantages of the Invention

[0018] The composition for enhancing the effect of the lactic acid bacterium of the present invention can enhance the physiological activity functions of the lactic acid bacterium, such as the effect of promoting the individual growth of the lactic acid bacterium or the effect of inhibiting intestinal injury, by containing branched-chain amino acids.

[0019] The composition for enhancing the effects of lactic acid bacteria of the present invention can enhance the physiological activity function of lactic acid bacteria by containing branched-chain amino acids synthesized by the synthase of branched-chain amino acids lacking in lactic acid bacteria.

[0020] The pharmaceutical composition or food composition of the present invention can promote the growth promotion effect and / or intestinal injury improvement effect of an individual by containing the composition for enhancing the effects of lactic acid bacteria together with lactic acid bacteria.

Brief Description of the Drawings

[0021]

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Mode for Carrying Out the Invention

[0022] Hereinafter, the present invention will be described in detail.

[0023] The present invention provides a composition for enhancing the effects of lactic acid bacteria containing at least two branched-chain amino acids (BCAA) selected from the group consisting of leucine, isoleucine, and valine.

[0024] The term "effect of lactic acid bacteria" means the functions and effects of lactic acid bacteria acting on a specific individual, and can also be expressed as the "physiological activity function of lactic acid bacteria". The term "physiological activity" means the property of a specific drug or substance acting on an organism.

[0025] The composition for enhancing the effects of lactic acid bacteria may be for enhancing the growth promoting effect of lactic acid bacteria or for enhancing the intestinal injury inhibitory effect of lactic acid bacteria.

[0026] The effects of lactic acid bacteria may be to promote the growth of an individual or to inhibit intestinal injury. Specifically, the effects of lactic acid bacteria may be to increase at least one of the body weight or bone density of an individual, or to inhibit intestinal leakage of an individual.

[0027] The effects of lactic acid bacteria may be to increase at least one of the body weight or bone density of a nutritionally deficient individual, or to inhibit intestinal leakage of a nutritionally deficient individual.

[0028] The term "growth promoting effect of lactic acid bacteria" means the effect of lactic acid bacteria to promote the growth of other organisms or individuals. For example, when the composition for enhancing the effects of lactic acid bacteria of the present invention is used in combination with lactic acid bacteria, the growth promoting effect of lactic acid bacteria on the treated individual can be increased. Specifically, compared with the case where lactic acid bacteria are administered alone to an arbitrary individual, when the composition for enhancing the effects of the present invention is administered to an arbitrary individual in combination with lactic acid bacteria, the growth promoting effect of the individual is significantly superior. According to one embodiment, it was confirmed that, compared with the case where only lactic acid bacteria are treated, when lactic acid bacteria and the composition for enhancing the effects of lactic acid bacteria of the present invention are used in combination, a significantly superior bone density increase and body weight increase effect are shown in the individuals treated with these in combination. High bone density helps to increase height, and for ordinary technicians, bone density is utilized as an important indicator of growth.

[0029] The term "intestinal injury inhibitory effect of lactic acid bacteria" means the effect of lactic acid bacteria that inhibits intestinal injury in other organisms or individuals. For example, when the composition for enhancing the effect of lactic acid bacteria of the present invention is used in combination with lactic acid bacteria, in the individual treated with lactic acid bacteria, the intestinal injury inhibitory effect of lactic acid bacteria on the individual can be increased. Specifically, compared with the case where lactic acid bacteria are administered alone to an arbitrary individual, when the composition for enhancing the effect of the present invention is administered to an arbitrary individual in combination with lactic acid bacteria, the intestinal injury inhibitory effect of the individual is significantly superior. Intestinal injury may be, but is not limited to, intestinal leakage and intestinal inflammation.

[0030] According to one embodiment, it was confirmed that, compared with the case where only lactic acid bacteria were treated, when lactic acid bacteria and the composition for enhancing the effect of lactic acid bacteria of the present invention were used in combination, a significantly superior intestinal leakage inhibitory effect was shown in the individual treated with these in combination.

[0031] The composition for enhancing the effect of lactic acid bacteria can serve as an adjuvant that can enhance the effects of lactic acid bacteria (for example, growth promoting effect, intestinal injury inhibitory effect).

[0032] The composition for enhancing the effect of lactic acid bacteria may be used in combination with lactic acid bacteria, and can serve as an adjuvant that can enhance the effect of lactic acid bacteria alone when used in combination with lactic acid bacteria.

[0033] The composition for enhancing the effect of lactic acid bacteria can be treated simultaneously, separately, or sequentially with lactic acid bacteria.

[0034] The individual may be an animal including a human, for example, a human, a dog, a cat, a horse, a cow, a rabbit, a goat, but is not limited thereto.

[0035] The individual may be an individual in a state of nutritional deficiency or nutritional imbalance. For example, the individual may be an individual lacking in protein nutrition.

[0036] The composition for enhancing the effects of lactic acid bacteria can, when used in combination with lactic acid bacteria, increase at least one of the body weight or bone density of an organism or individual with nutritional deficiency or nutritional imbalance, or can be used to inhibit intestinal leakage in nutritionally deficient individuals. "Nutritional deficiency" can mean a state in which one or more essential nutrients or calories are lacking, and can occur, for example, due to insufficient intake of nutrients, absorption or processing disorders, etc. In the case of nutritionally deficient individuals, symptoms such as underweight, protruding bones, dry and less elastic skin, or dry hair may appear. "Nutritional imbalance" means an imbalance between the nutrients required by the body and the nutrients obtained from the body, and can include both overnutrition and undernutrition.

[0037] The composition for enhancing the effects of lactic acid bacteria can enhance the effect of lactic acid bacteria in promoting the growth of individuals with nutritional deficiency or nutritional imbalance, or can enhance the effect of inhibiting intestinal damage.

[0038] The age of the individual is not limited, and can be, for example, an adult excluding children and the elderly, or an elderly person.

[0039] The individual can be an individual in childhood. "Childhood" means the period from fertilization through birth to adulthood. Childhood can include the cellular stage, fetal stage, neonatal stage, infancy, childhood, and adolescence.

[0040] The composition for enhancing the effects of lactic acid bacteria can enhance the activity of lactic acid bacteria in vitro. For example, the composition for enhancing the effects of lactic acid bacteria can promote the growth of cells, tissues, etc. that are treated with lactic acid bacteria in vitro. For example, the composition for enhancing the effects of lactic acid bacteria can inhibit the damage of intestinal-related cells and tissues that are treated with lactic acid bacteria in vitro.

[0041] Lactic acid bacteria are bacteria that utilize sugar as an energy source to produce lactic acid. They exist in the digestive organs and oral cavities of humans and mammals, and are widely distributed in nature such as various fermented foods. Lactic acid bacteria are one of the microorganisms that humans have used widely for the longest time. They are microorganisms that do not produce substances harmful to the intestines of humans and animals and have the function of preventing intestinal putrefaction. Lactic acid bacteria mean beneficial bacteria that can produce lactic acid.

[0042] The lactic acid bacteria may be Lactobacillaceae, Enterococcaceae, Streptococcaceae, or Bifidobacteriaceae.

[0043] The lactic acid bacteria may be selected from the group consisting of Lactobacillus spp., Lactococcus spp., Enterococcus spp., Streptococcus spp., and Bififobacterium spp.

[0044] The genus Lactobacillus may be selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus rhamnosus, and Lactobacillus salivarius, but is not limited thereto.According to one embodiment, when the composition of the present invention is used in combination with at least one lactic acid bacterium selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius, the effect of the lactic acid bacterium can be enhanced.

[0045] The lactic acid bacterium may be isolated from plants, fermented foods, animal individuals including mammals and arthropods, or may be present in the intestine of an individual. The composition for enhancing the effect of the lactic acid bacterium of the present invention can also enhance the action effect of the isolated lactic acid bacterium and can also enhance the effect of the lactic acid bacterium present in the intestine of an individual.

[0046] The lactic acid bacterium may be a live bacterium or a dead bacterium, and its form is not limited.

[0047] L. plantarum (L. plantarum WJL) used in one embodiment may be produced by a known production method (Eun-Kyoung Kim et al., Genome Announcements, November / December 2013, Vol.1, no.6 e00937-13, GenBank AUTE00000000, Lactobacillus plantarum WJL, whole genome shotgun sequencing project). L. plantarum (L. plantarum WJL) may be isolated from Drosophila melanogaster. L. plantarum (L. plantarum WJL) may be a deposited strain (Accession number: KCTC14442BP, Depositary institution: Biological Resource Center, Date of deposit: January 11, 2021).

[0048] The inventors considered that by using branched-chain amino acids in combination with lactic acid bacteria in view of the fact that lactic acid bacteria lack genes related to branched-chain amino acid biosynthetic enzymes and cannot synthesize branched-chain amino acids due to gene analysis of lactic acid bacteria, the effects of lactic acid bacteria can be enhanced, and thus they were selected as the active ingredient of the composition for enhancing the effects of the lactic acid bacteria of the present invention. The composition for enhancing the effects of lactic acid bacteria can exert the effect of enhancing the efficacy of lactic acid bacteria by using in combination the amino acids that cannot be biosynthesized by lactic acid bacteria.

[0049] The term "branched-chain amino acid (BCAA)" refers to an amino acid having an aliphatic side chain with a branch that is a central carbon atom bonded to three or more carbon atoms. Proteinogenic BCAA includes the essential amino acids leucine, isoleucine, and valine, and non-proteinogenic BCAA includes 2-aminoisobutyric acid. The branched-chain amino acid may be a free amino acid. "Free amino acid" means an amino acid that is not part of a protein.

[0050] The composition for enhancing the effect of lactic acid bacteria can contain at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine, and valine.

[0051] Specifically, the composition for enhancing the effect of lactic acid bacteria can contain leucine and isoleucine; leucine and valine; isoleucine and valine; or leucine, isoleucine, and valine.

[0052] When the composition for enhancing the effect of lactic acid bacteria of the present invention contains all two or more of leucine, isoleucine, and valine, the effect of lactic acid bacteria can be enhanced more effectively than when they are contained alone.

[0053] The composition for enhancing the effect of lactic acid bacteria (for example, the composition for enhancing the growth promoting effect of lactic acid bacteria or the composition for enhancing the intestinal injury inhibitory effect of lactic acid bacteria) can contain leucine, isoleucine, and valine.

[0054] When the composition for enhancing the effect of lactic acid bacteria of the present invention contains all of leucine, isoleucine, and valine, the activity of lactic acid bacteria can be enhanced most effectively.

[0055] According to one embodiment, compared with the case where a composition containing leucine, isoleucine, or valine alone is treated in combination with lactic acid bacteria, when a composition containing two or more of leucine, isoleucine, and valine is treated in combination with lactic acid bacteria, it can be confirmed that the growth of the individuals treated therewith is significantly enhanced.

[0056] The branched-chain amino acids contained in the composition of the present invention are not merely included for individual nutritional supplementation, but can play a role in improving the function of lactic acid bacteria in the intestine.

[0057] As described above, when using branched-chain amino acids lacking synthesis in lactic acid bacteria in combination with lactic acid bacteria, the physiological activities of lactic acid bacteria (for example, growth promotion effect or intestinal injury inhibition effect) are enhanced, and even without using other proteins other than branched-chain amino acids, the composition of the present invention alone can exhibit a sufficient effect enhancement effect. Therefore, the composition for enhancing the effect of lactic acid bacteria of the present invention has an excellent effect of enhancing the effect of lactic acid bacteria and is also excellent economically from the viewpoint of raw material consumption.

[0058] For example, the composition for enhancing the effect of lactic acid bacteria may not contain other proteins (for example, whey protein) other than branched-chain amino acids. The composition for enhancing the effect of lactic acid bacteria can significantly enhance the effect of lactic acid bacteria without containing other proteins (for example, whey protein) other than the above. In this case, the raw material cost of other proteins can be reduced, which is economically excellent. In addition, by containing other proteins, the possibility of functional decline of branched-chain amino acids and unexpected side effects can be reduced.

[0059] "Whey protein" is milk protein from which casein has been removed. 80% of whey protein consists of lactalbumin and lactoglobulin. In addition, it contains components such as proteose and peptone that do not coagulate by acids or heat. Whey protein has the advantage of showing effects such as improving the recovery rate after exercise, satiety, and weight control. However, individuals with lactose intolerance lacking lactase or individuals with milk allergy should be careful because symptoms such as diarrhea and abdominal pain may appear when ingesting whey protein. In addition, individuals with kidney or liver diseases who need to restrict protein intake should also be careful about ingesting whey protein.

[0060] The composition for enhancing the effect of lactic acid bacteria of the present invention may not contain whey protein. Therefore, there are no problems (such as diarrhea, abdominal pain, allergy, etc.) that may occur when whey protein is contained, and at the same time, the growth promotion or intestinal injury inhibition effect of lactic acid bacteria can be enhanced.

[0061] In other examples, the composition for enhancing the effect of lactic acid bacteria may consist of at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine, and valine.

[0062] The present invention also provides a method for enhancing the effect of lactic acid bacteria, which includes the step of administering the aforementioned composition for enhancing the effect of lactic acid bacteria to an individual.

[0063] Since the "composition for enhancing the effect of lactic acid bacteria", "lactic acid bacteria", and "effect of lactic acid bacteria" are as described above, specific explanations are omitted.

[0064] The lactic acid bacteria may be present in the intestine of the individual.

[0065] The individual may be an animal including a human, for example, a human, a dog, a cat, a horse, a cow, a rabbit, a goat, but is not limited thereto.

[0066] The individual may be an individual with inhibited growth or an individual with intestinal injury.

[0067] The individual may be an individual with inhibited growth or an individual with intestinal injury due to nutritional deficiency or nutritional imbalance.

[0068] The method for enhancing the effect of lactic acid bacteria may not include administering a polypeptide or a protein (for example, whey protein) in addition to the branched-chain amino acids and lactic acid bacteria.

[0069] The present invention also provides a pharmaceutical composition for promoting growth or inhibiting intestinal injury, which includes the aforementioned composition for enhancing the effect of lactic acid bacteria and lactic acid bacteria.

[0070] The pharmaceutical composition of the present invention includes all of the composition for enhancing the effect of lactic acid bacteria that enhances the growth-promoting effect and intestinal injury-inhibiting effect of lactic acid bacteria and lactic acid bacteria, so that it can promote the growth of the individual to whom the pharmaceutical composition is administered or inhibit intestinal injury.

[0071] Since the "composition for enhancing the effects of lactic acid bacteria" and "lactic acid bacteria" have been described above, specific descriptions thereof are omitted.

[0072] The growth promoting effect may mean the effect of promoting the growth of an individual including a human to whom the pharmaceutical composition has been administered.

[0073] Growth promotion may be at least one of an increase in body weight, an increase in height, and an increase in bone density.

[0074] According to one embodiment, it was confirmed that body weight and bone density increased in an animal in which the composition for enhancing the effects of lactic acid bacteria of the present invention and lactic acid bacteria were used in combination.

[0075] The intestinal injury inhibitory effect may mean the effect of inhibiting intestinal injury of an individual including a human to whom the pharmaceutical composition has been administered.

[0076] Intestinal injury may be, but is not limited to, inflammatory bowel disease, environmental enteropathy, or intestinal leakage syndrome.

[0077] According to one embodiment, it was confirmed that intestinal leakage was inhibited in an animal to which the composition for enhancing the effects of lactic acid bacteria of the present invention and lactic acid bacteria were administered in combination.

[0078] The present invention also provides a composition for preventing or treating at least one disease selected from the group consisting of growth disorders, growth retardation, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal leakage syndrome, which comprises the aforementioned composition for enhancing the effects of lactic acid bacteria and lactic acid bacteria. This may be an effect in which the physiological activity effects of lactic acid bacteria (for example, growth promoting effect and intestinal injury inhibitory effect) are enhanced and appear in an individual to whom the pharmaceutical composition has been administered.

[0079] The term "prevention" means all acts of suppressing a disease or delaying its progression by administering the composition for enhancing the effects of lactic acid bacteria and lactic acid bacteria. The term "treatment" means all acts in which the symptoms of a disease are improved or beneficially changed by administering the composition for enhancing the effects of lactic acid bacteria and lactic acid bacteria.

[0080] The disease may be caused by nutritional deficiency or imbalance. Since "nutritional deficiency or imbalance" is as described above, specific description is omitted.

[0081] The disease may occur in animals including humans, and its age is not limited.

[0082] The disease may occur in individuals other than children and the elderly, specifically adults excluding the elderly, or in elderly individuals. Specifically, it may also be a disease that occurs in individuals in childhood. Since "childhood" is as described above, specific description is omitted.

[0083] The pharmaceutical composition of the present invention can be administered to individuals with nutritional deficiency or imbalance.

[0084] The pharmaceutical composition of the present invention can be administered to individuals in childhood.

[0085] The pharmaceutical composition of the present invention may be for preventing or treating at least one disease selected from the group consisting of growth disorders, stunted growth, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal permeability syndrome in individuals in childhood.

[0086] Intestinal permeability syndrome is a disease that occurs when spaces form between the cells that line the inner wall of the intestine, resulting in reduced intestinal function, increased intestinal permeability, and inducing disorders in the absorption of water and nutrients and the immune system.

[0087] Environmental enteropathy (EE) is a disease in which malnutrition and intestinal infections occur simultaneously, causing stunted growth and reduced intelligence in children. It is not easy to improve environmental enteropathy with just nutritional supply alone.

[0088] The branched-chain amino acids contained in the pharmaceutical composition of the present invention are not for the sole purpose of nutritional supply, but can improve the effects of lactic acid bacteria and show excellent effects in improving environmental enteropathy.

[0089] According to one embodiment, the pharmaceutical composition of the present invention comprises at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine and valine, and at least one lactic acid bacterium selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus and Lactobacillus salivarius.

[0090] According to one embodiment, a pharmaceutical composition comprising at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine, and valine, and at least one lactic acid bacterium selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius can increase the body weight and bone density of an individual to be administered.According to one embodiment, a pharmaceutical composition comprising at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine and valine, and at least one lactic acid bacterium selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus and Lactobacillus salivarius can inhibit intestinal leakage in an individual to be administered.

[0091] The pharmaceutical composition of the present invention may not contain other proteins (e.g., whey protein) in addition to branched-chain amino acids and lactic acid bacteria, and even without containing the other proteins (e.g., whey protein), the branched-chain amino acids and lactic acid bacteria can significantly promote the growth of the individual to which they are administered or significantly inhibit intestinal injury. In this case, costs such as those of other proteins (e.g., whey protein) can be reduced, which is economically advantageous.

[0092] The pharmaceutical composition of the present invention can further contain a pharmaceutically acceptable carrier, excipient or diluent. Pharmaceutically acceptable carriers are those commonly used in formulation, for example, physiological saline, sterile water, Ringer's solution, buffered physiological saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, antioxidants, but are not limited thereto.

[0093] The pharmaceutical composition of the present invention can be formulated into injectable preparations, pills, capsules, granules or tablets, but is not limited thereto.

[0094] The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) by the intended method and can be appropriately selected by those skilled in the art.

[0095] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

[0096] The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or multiply.

[0097] The effective amount of the pharmaceutical composition of the present invention varies depending on the age, sex, condition, weight of the patient, absorption degree, inactivation rate and excretion rate of the active ingredient in the body, type of disease, and drugs used in combination. Generally, it can be administered at 0.001 to 150 mg, preferably 0.01 to 100 mg per kg of body weight daily or every other day, or divided into 1 to 3 times a day. However, since it can be increased or decreased depending on the administration route, severity of obesity, sex, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0098] In addition, the present invention provides a method for promoting growth or inhibiting intestinal injury, which includes the steps of administering a composition for enhancing the effect of lactic acid bacteria and lactic acid bacteria to an individual.

[0099] Since the "composition for enhancing the effects of lactic acid bacteria", "lactic acid bacteria", "growth promotion", and "intestinal injury inhibition" are as described above, specific explanations are omitted.

[0100] The individual may be an animal including humans, for example, it may be a human, dog, cat, horse, cow, rabbit, or goat, but is not limited thereto.

[0101] The individual may be an individual with inhibited growth or an individual with intestinal injury.

[0102] The individual may be an individual with inhibited growth or an individual with intestinal injury due to nutritional deficiency or imbalance.

[0103] The composition for enhancing the effects of lactic acid bacteria and lactic acid bacteria can be administered simultaneously, separately, or sequentially.

[0104] The method for promoting growth or inhibiting intestinal injury may not include administering a polypeptide or protein (e.g., whey protein) in addition to branched-chain amino acids and lactic acid bacteria.

[0105] The present invention also provides a food composition for promoting growth or inhibiting intestinal injury, comprising the aforementioned composition for enhancing the effects of lactic acid bacteria and lactic acid bacteria.

[0106] The food composition of the present invention includes all of the composition for enhancing the effects of lactic acid bacteria that enhances the growth-promoting effect and intestinal injury-inhibiting effect of lactic acid bacteria and lactic acid bacteria, thereby promoting the growth of the individual to whom the food composition is administered or inhibiting intestinal injury.

[0107] Since the "composition for enhancing the effects of lactic acid bacteria", "lactic acid bacteria", "growth promotion", "intestinal injury inhibition", and "intestinal injury" are as described above, specific explanations are omitted.

[0108] The food composition of the present invention may be used for preventing or improving at least one disease selected from the group consisting of growth disorders, stunted growth, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal leakage syndrome. This may be an effect in which the effects of lactic acid bacteria (for example, growth promoting effect and intestinal injury inhibiting effect) are enhanced and manifested in an individual administered with the food composition.

[0109] The term "improvement" means all actions that reduce parameters related to the condition treated by the intake of the food composition, for example, the degree of symptoms of the disease.

[0110] Since the "disease" is as described above, specific description is omitted.

[0111] The food composition of the present invention can be administered to individuals with nutritional deficiencies or imbalances.

[0112] The food composition of the present invention can be administered to individuals in childhood.

[0113] The food composition of the present invention may be used for preventing or improving at least one disease selected from the group consisting of growth disorders, stunted growth, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal leakage syndrome in individuals in childhood.

[0114] According to one embodiment, the food composition of the present invention can include at least two branched-chain amino acids selected from the group consisting of leucine, isoleucine, and valine, and at least one lactic acid bacterium selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius.

[0115] In addition to branched-chain amino acids and lactic acid bacteria, the food composition of the present invention may not contain other proteins (e.g., whey protein), and even without containing the other proteins (e.g., whey protein), the branched-chain amino acids and lactic acid bacteria can significantly promote the growth of the individual to whom they are administered or significantly inhibit intestinal injury. In this case, costs such as those of other proteins (e.g., whey protein) can be reduced, which is economically superior.

[0116] The food composition can be manufactured into various forms of preparations without limitation as long as they are preparations recognized as foods. For example, foods include meat, sausages, bread, chocolate, candies, snacks, confectioneries, pizza, ramen, other noodles, gums, dairy products including ice creams, various soups, beverages, tea, drink agents, alcoholic beverages, vitamin complexes, health functional foods, and health foods, etc., and all foods in the ordinary sense are included.

[0117] The food composition can be manufactured by methods commonly used in the art.

[0118] The food composition can further contain raw materials and components commonly added in the art. For example, the food composition can further contain vitamin A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid. For example, the food composition can further contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), chromium (Cr).

[0119] Hereinafter, the configuration and effects of the present invention will be described more specifically with examples. However, the following examples are provided for illustrative purposes to facilitate the understanding of the present invention and do not limit the scope and range of the present invention.

Example

[0120] The inventors derived a combination of lactic acid bacteria and branched-chain amino acids that enhance the effects of lactic acid bacteria as an active ingredient for satisfying the nutrition of nutritionally deficient individuals through the following experiments. Prior to the results regarding whether lactic acid bacteria and branched-chain amino acids that enhance the effects of lactic acid bacteria exhibit the effect of enhancing the efficacy of lactic acid bacteria, the specific indicators appearing in nutritionally deficient individuals and the process of selecting lactic acid bacteria as one component for satisfying the nutrition of nutritionally deficient individuals will be explained.

[0121] To confirm whether the effect of enhancing the efficacy of lactic acid bacteria is statistically significant by comparing the control group and the experimental group, statistical analysis was performed using one-way ANOVA (with Dunnett's multiple comparison post-test). p value: *** (p < 0.001), ** (P < 0.01), * (P < 0.05), statistically significant results were shown. Experimental groups with high p values and no statistically significant results were labeled as "ns".

[0122] 1. Confirmation of specific indicators appearing in nutrient-deficient individuals 1-1. L-EAA preference in nutrient-deficient individuals Based on the feeding behavior of Drosophila that specifically ingests essential amino acids in a situation where the body protein is deficient, the behavioral indicators of nutrient-deficient individuals were observed. Specifically, Drosophila were placed in a medium without protein deficiency and a medium with protein deficiency (a medium containing only glucose), and it was confirmed which type of food they preferred among the foods of L-form essential amino acids (the form of amino acids existing in nature, L-EAA) and D-form essential amino acids (optical isomers of L-form essential amino acids, D-EAA).

[0123] As a result, it was confirmed that the longer the individuals were exposed to the protein-deficient medium, the higher the feeding preference index for L-EAA (see FIGS. 1A and 1B). In addition, in nutrient-deficient individuals, it was confirmed that they preferred to ingest L-type essential amino acids among L-type amino acids, and that the feeding preference for L-form nonessential amino acids (L-NEAA) did not increase even in the situation of protein deficiency (see FIGS. 1C to 1D). In addition, as a result of comparing the L-EAA feeding preference of Drosophila individuals fed diets with the same calorie content but different protein ratios, it was confirmed that the feeding preference for L-EAA decreased in individuals fed a diet with a high protein content. From this, it was confirmed that the feeding behavior of Drosophila to supplement L-EAA is finely regulated to meet the protein nutrition (see FIG. 1E). Furthermore, after inhibiting the activity of Ir76b cells, which have conventionally been known as taste receptors that recognize proteins, the L-EAA feeding preference was confirmed. Specifically, the L-EAA feeding preference was confirmed by placing Drosophila with overexpression of TNT (tetanus toxin) to inactivate Ir76b cells and control group Drosophila in a protein-deficient situation. As a result, Drosophila with inactivated Ir76b cells showed an L-EAA feeding preference at a level similar to that of the control group Drosophila (see FIG. 1F). From this, it was found that there is another signal transduction pathway other than the conventionally known signal transduction pathway that recognizes in-body nutrient deficiency.

[0124] 1-2. Overexpression of CNMa hormone in nutrient-deficient individuals To investigate a new signaling pathway that recognizes nutritional deficiencies in the body, we focused on the intestine, an endocrine organ where food is digested and absorbed and various hormones are secreted. Drosophila were fed a medium lacking protein and a medium containing protein, and their intestines were dissected to confirm the mRNA expression levels of 11 intestinal hormones under each condition by real-time qPCR. The primers used for real-time qPCR are shown in Table 1 below.

[0125]

Table 1

[0126] As a result, in individuals under nutrient-deficient conditions, the expression level of CNMa among the 11 intestinal hormones in Table 1 above increased. From this, it was confirmed that protein deficiency induces the CNMa hormone (see Figure 2A). Also, flies were provided with media having the same calorie content but different protein (yeast) contents, and the expression level of CNMa mRNA in the intestines of the flies under each media condition was measured. As a result, it was confirmed that the mRNA expression of the CNMa hormone decreases as the protein content in the media increases (see Figure 2B). To confirm the expression level of the CNMa hormone in vivo, Gal4 lines (CNMa-Gal4) of flies in which expression is regulated by the CNMa promoter were created. Specifically, flies of CNMa[MI10321] (BDSC#54529) in which Mimic is inserted into the first intron part of the CNMa gene were used with the recombinase-mediated cassette exchange (RMCE) system to replace Mimic with gal4 to create CNMa-gal4. As a result of confirming the CNMa expression of the created fly line with GFP, indeed, in the intestines of the flies, when the amount of protein ingested by the flies is small, the expression of the CNMa hormone increases significantly, and when the flies ingest a high-protein diet, the expression of the CNMa hormone decreases rapidly (see Figures 2C and 2D). In particular, in the intestines of the flies, it was confirmed that the expression of CNMa specifically increases in the enterocytes in the R2 region (see Figures 2C and 2D). Also, to confirm the expression of endogenous CNMa, a CNMa-specific antibody was created and immunohistochemistry was performed. As a result, similar to the expression of the CNMa-Gal4 line, it was reconfirmed that the CNMa hormone is expressed in the enterocytes in the R2 region of the intestines of the flies (see Figure 3). From the above experimental results, it was confirmed that the deficiency of essential amino acids in the body can be recognized from the expression level of the CNMa hormone in an individual.

[0127] To confirm whether the lack of specific types of essential amino acids induces the expression of CNMa hormone, the researchers provided Drosophila with media lacking arginine, tryptophan, valine, isoleucine, or leucine alone. As a result, it was confirmed that the lack of a single essential amino acid, rather than the lack of specific essential amino acids, induces the expression of CNMa hormone in intestinal epithelial cells (see Figure 4A). Similarly, it was confirmed that the lack of a single essential amino acid alone induces an increase in the feeding preference for L-EAA in Drosophila (see Figure 4B). This indicates that Drosophila becomes nutritionally deficient even with the lack of a single essential amino acid.

[0128] 2. Confirmation of the effects of gut microbiota on individuals To confirm the effect of gut microbiota on nutritional sufficiency in individuals, the following experiments were conducted.

[0129] Eggs laid by conventionally reared flies (CV) were collected, rinsed alternately with 3% NaClO solution and 70% ethanol solution to remove the bacteria contained in the eggs, and aseptic Drosophila were created by inducing development in aseptic medium. The preference for L-EAA in the germ-free fly (GF) thus created was confirmed in media lacking and not lacking protein. As a result, germ-free Drosophila showed an increased basal L-EAA preference compared to conventionally reared flies (CV) even under conditions of sufficient protein, and the feeding preference increased with time. That is, it was confirmed that germ-free Drosophila always felt nutrient deficiency (see Figure 5A). In the case of germ-free Drosophila, since they showed a feeding behavior preferring essential amino acids even in the absence of protein deficiency, it was found that gut bacteria play an important role in offsetting L-EAA deficiency.

[0130] Furthermore, after feeding normal Drosophila and axenic Drosophila with a diet containing 10% yeast, the expression level of intestinal CNMa was examined. As a result, it was confirmed that the expression of CNMa hormone in axenic Drosophila increased, despite the same protein content in the medium. When yeast was further added to the medium (Yeast 15%), it was confirmed that the increased expression of CNMa decreased (see Fig. 5B). It was confirmed that the preference for L-EAA ingestion in axenic Drosophila increased compared to normal Drosophila, despite the same protein content in the medium. Furthermore, when yeast was further added to the medium (Yeast 15%), the preference for L-EAA ingestion in axenic Drosophila decreased (see Fig. 5C). From these results, it was confirmed that gut microbiota counteracts L-EAA deficiency and affects the expression of CNMa hormone in Drosophila and the behavior to satisfy L-EAA nutrition.

[0131] 3. Confirmation of the nutrient sufficiency effect according to the types of gut microbiota introduced Among intestinal microorganisms, the most representative Acetobacter pomorum (A. pomorum) (Science, February 8, 2008; 319(5864): 777-82. doi: 10.1126 / science.1149357. Epub January 24, 2008; Appl Environ Microbiol. October 2008; 74(20): 6171-6177. Published online August 22, 2008. doi: 10.1128 / AEM.00301-08) and Lactobacillus plantarum (L. plantarum WJL) (KCTC14442BP) were introduced into germ-free Drosophila melanogaster respectively to create mono-associated Drosophila melanogaster, and then the expression of intestinal CNMa hormone in each individual was confirmed. As a result, it was confirmed that the expression level of CNMa hormone was low in Drosophila melanogaster introduced with A. pomorum, while the expression level of CNMa hormone in Drosophila melanogaster introduced with L. plantarum WJL was as high as that in germ-free Drosophila melanogaster (see Figure 6A). L. plantarum WJL described in the description of the present invention may be produced by a known production method (Eun-Kyoung Kim et al., Genome Announcements, November / December 2013, Vol.1, no. 6 e00937-13, GenBank AUTE00000000, Lactobacillus plantarum WJL, whole genome shotgun sequencing project). L. plantarum WJL may be isolated from Drosophila melanogaster.

[0132] In addition, when the L-EAA nutritional sufficiency behavior index was examined in each individual, it was confirmed that only the Drosophila melanogaster into which A. pomorum was introduced decreased the L-EAA sufficiency behavior index (see Fig. 6B). Furthermore, when L. plantarum WJL was introduced alone, the result showed that the feeding preference was higher than that of germ-free Drosophila melanogaster, which means that L. plantarum WJL can exacerbate the deficiency of essential amino acids in the intestine of the host animal. This corresponds to the expression pattern of the CNMa hormone.

[0133] From the results that the expression level of the CNMa hormone and the L-EAA feeding preference were high in the individuals into which the intestinal microorganism Lactobacillus plantarum was introduced, the present researchers were able to predict that the nutritional sufficiency effect would not be exerted even if Lactobacillus plantarum alone was administered to the nutrient-deficient individuals. Therefore, in order to investigate the reasons why the Drosophila melanogaster into which A. pomorum was introduced and the Drosophila melanogaster into which L. plantarum WJL was introduced showed different degrees of CNMa expression and L-EAA feeding preference, the genes of Lactobacillus and lactic acid bacteria similar thereto were analyzed.

[0134] 4. Gene analysis of gut microbiota and the effect difference due to the genetic traits of gut microbiota 4-1. Gene analysis of Acetobacteraceae and Lactobacillaceae By comparative genomic analysis, genes related to amino acid biosynthesis in Acetobacteraceae and Lactobacillaceae were compared. As a result of the interpretation, it was confirmed that Acetobacteraceae has all the enzymes related to the biosynthesis of branched-chain amino acids (BCAAs), while Lactobacillaceae lacks the enzymes related to the biosynthesis of BCAAs (see Fig. 6C). Using the genome of the Acetobacter pomorum DM001 (A. pomorum DM001) strain, which is capable of synthesizing all amino acids, as a template, the higher the nucleotide sequence similarity of the genes, the more they were displayed in blue, and those with a similarity of less than 20% were displayed in red.

[0135] 4-2. Confirmation of the lack of BCAA synthesis-related genes in various species of lactic acid bacteria In order to confirm whether BCAA synthesis-related genes are lacking in various lactic acid bacteria other than Lactobacillus plantarum, the following experiments were conducted.

[0136] Lactobacillus plantarum (KCTC14442BP), Lactobacillus acidophilus (ATCC 4796), Lactobacillus casei (ATCC 393), Lactobacillus gasseri (ATCC33323), Lactobacillus bulgaricus (ATCC 11842), Lactobacillus helveticus (ATCC 15009), Lactobacillus fermentum (ATCC 14931), Lactobacillus paracasei (ATCC 334), Lactobacillus reuteri (JCM 1112), Lactobacillus rhamnosus (ATCC 8530), Lactobacillus salivarius (DSM 20555), which are lactic acid bacteria (strains of Food Notice) widely used in foods and lactic acid bacteria products, were targeted, and enzymes related to amino acid biosynthesis were identified by comparative genomic analysis. The genomes of A. pomorum strains capable of synthesizing all amino acids were used as templates for comparison (see Figure 7). As a result of comparative genomic analysis, it was confirmed that all 11 Lactobacillus strains lacked leuA, leuB, leuC, leuD, ilvA, ilvC, ilvD, or ilvE, which are genes related to BCAA biosynthesis (see Figure 7).

[0137] From these results, it was found that in the case of Lactobacillus plantarum, due to its genetic traits, even if it is introduced into an individual, it does not show a nutritional sufficiency effect.

[0138] 4-3. Proof that the nutritional sufficiency effect on an individual can vary depending on the genetic traits of the intestinal microorganisms to be introduced To clarify that the genetic traits of gut microbiota can induce host intestinal L-EAA deficiency (CNMa hormone expression), as follows, an Acetobacteraceae with a mutation in the gene related to BCAA synthesis was created, and the host intestinal L-EAA deficiency was confirmed. The mutant strain of a specific gene of Acetobacter was prepared by amplifying the regions on both sides of the gene to be deleted by PCR and inserting them into the pK18mobGII vector, and the cloned vector was introduced into the wild-type Acetobacter strain by the triparental mating method together with a helper strain (Escherichia coli HB101).

[0139] Specifically, a mutant Acetobacter (Acto ΔleuB ) that cannot biosynthesize leucine, one of the BCAA, was created. In the Drosophila melanogaster introduced with the control group of Acetobacter (+Aceto WT ), the CNMa hormone was hardly expressed. In contrast, in the Drosophila melanogaster introduced with Aceto ΔleuB (+Aceto ΔleuB ), the expression of the CNMa hormone increased to the level of axenic Drosophila melanogaster.

[0140] Furthermore, a proC mutant Acetobacter (Aceto ΔproC ) that cannot biosynthesize proline, one of the non-essential amino acids, was created. Different from the case of introducing Aceto ΔleuB into axenic Drosophila melanogaster, in the case of Drosophila melanogaster introduced with Aceto ΔproC bacteria (+Aceto ΔproC ), the expression of the CNMa hormone did not increase (see the left side of Fig. 8A).

[0141] On the other hand, leucine was added to the medium of Drosophila melanogaster introduced with Aceto ΔleuB (+Aceto ΔleuB +Leucine), or Aceto ΔleuB bacteria were genetically re-introduced with leuB to obtain AcetoΔleuB_leuB Flies introduced with bacteria (+Aceto ΔleuB_leuB ) showed a decrease in the expression of the CNMa hormone (see the right side of Fig. 8A). Similar to the expression pattern of the CNMa hormone, the behavior of the germ-free flies as the host to satisfy L-EAA nutrition also increased when bacteria with Aceto WT introduced was compared with the case where Aceto ΔleuB bacteria was introduced (see Fig. 8B). The behavior of flies introduced with Aceto ΔleuB to satisfy L-EAA nutrition decreased when leucine was added to the medium (+Aceto ΔleuB +Leucine), or when flies were introduced with bacteria genetically reintroduced with the deficient leuB (+Aceto ΔleuB_leuB ) (see Fig. 6B). This means that the leucine biosynthesis ability of Acetobacter plays an important role in the expression of the CNMa hormone in intestinal epithelial cells and the L-EAA feeding behavior of the host flies.

[0142] Similarly, an Aceto ΔilvA mutant strain that cannot biosynthesize isoleucine among BCAA was created, and the same experiment as above was conducted (see Fig. 9). As a result of comparing the CNMa hormone expression levels and the behavioral indicators of the host to satisfy L-EAA nutrition among germ-free flies (+None), flies introduced with the control group of Acetobacter (+Aceto WT ), and flies introduced with Aceto ΔilvA that cannot biosynthesize isoleucine (+Aceto ΔilvA ), flies introduced with Aceto ΔilvA showed a behavior similar to that of germ-free flies (+None) in terms of CNMa expression level and satisfying L-EAA nutrition (see Fig. 9A). Also, when isoleucine was added to the medium of flies introduced with Aceto ΔilvA (+Aceto ΔilvA +Ile), the expression of CNMa and the behavior of satisfying L-EAA nutrition were similar to those of Aceto ΔleuB +Aceto WTIt was confirmed that the flies were restored to the level of Drosophila into which it was introduced (see Fig. 9B).

[0143] Furthermore, by introducing seven enzymes related to BCAA biosynthesis into Lactobacillus lacking genes related to BCAA biosynthesis, a gain-of-function mutant strain capable of synthesizing BCAA was created. The Lacto strain that genetically acquired the ability to synthesize BCAA amplified the seven genes of leuA, leuB, leuC, leuD, ilvA, ilvC, and ilvD required for the synthesis of BCAA using the genome of Lactobacillus coryniformis subsp. Torquens DSM 20004 strain as a template and inserted it into the PGID023A / B vector. This cloned vector was introduced into the Lacto strain using electroporation and inserted into a specific gene region by homologous recombination. BCAA The expression level of the CNMa hormone and the behavior of trying to satisfy L-EAA nutrition were observed in germ-free Drosophila (+None), Drosophila into which the control group of Lactobacillus was introduced (+Lacto WT ), and Drosophila into which the mutant Lactobacillus (Lacto

[0144] ) that genetically acquired the ability to synthesize BCAA was introduced (+Lacto WT ). As a result, it was confirmed that the expression of the CNMa hormone and the behavioral index of trying to satisfy L-EAA nutrition decreased in Drosophila into which Lacto BCAA was introduced compared to Drosophila into which Lacto BCAA was introduced (see Fig. 10A). Also, when BCAA (isoleucine, leucine, valine) was added to the medium of Drosophila into which Lacto BCAA was introduced, the expression of CNMa and the behavioral index of trying to satisfy L-EAA nutrition were Lacto WT compared to those of Drosophila into which Lacto WT was introduced. When BCAA (isoleucine, leucine, valine) was added to the medium of Drosophila into which Lacto BCAAIt was confirmed that the level decreased in the introduced Drosophila (see Fig. 10B).

[0145] From the above results, it was confirmed whether the genetic traits of intestinal microorganisms offset or did not offset the deficiency of L-EAA in the host's intestine in the nutritional deficiency situation of the host animal. In the case of Lactobacillus, which is a particularly representative lactic acid bacterium, it is difficult to synthesize BCAA and it cannot offset the deficiency of L-EAA in the host's intestine, indicating that it induces the feeding preference to satisfy the host's nutrition. It can be confirmed that when BCAA is supplemented here, the hormonal and behavioral changes (feeding preference) of the host caused by lactic acid bacteria disappear.

[0146] 5. Growth promotion and intestinal injury improvement effects of the combination of lactic acid bacteria and BCAA in nutrient-deficient individuals Based on the above experimental results, it was demonstrated that nutritionally deficient individuals have difficulty in supplementing nutrition even when they ingest lactic acid bacteria, and by concomitantly ingesting BCAA, which is not well synthesized by lactic acid bacteria when ingesting lactic acid bacteria, the nutritional requirements of the individuals are satisfied. Based on this result, it was considered that the effect of lactic acid bacteria alone can be enhanced when lactic acid bacteria and BCAA are treated together, and it was confirmed whether lactic acid bacteria can enhance the physiological activity effect of the host (the individual into which lactic acid bacteria were introduced) when lactic acid bacteria and BCAA are supplemented together in a nutritional deficiency situation, targeting the Drosophila animal model and the mouse animal model.

[0147] 5-1. Confirmation of the growth promotion effect by the combined treatment of lactic acid bacteria and BCAA in nutritionally deficient individuals 5-1-1. Effects of the combination of lactic acid bacteria and three types of BCAA, leucine, isoleucine, and valine Using a germ-free Drosophila model, growth changes were observed by separately or jointly adding Lactobacillus bacteria and BCAA (including all of leucine, isoleucine, and valine) to germ-free Drosophila in a protein-deficient medium (1% yeast, 10% sucrose, cornmeal medium). Specifically, in the group where BCAA (leucine, isoleucine, and valine) was added to germ-free Drosophila eggs (+BCAA); the group where L. plantarum WJL was introduced alone (+WJL); and the group where both L. plantarum WJL bacteria and BCAA (leucine, isoleucine, and valine) were introduced (+WJL+BCAA), the formation of pupae was observed every 12 hours, and the average period required for development (hereinafter, the average development period) was measured.

[0148] As a result, the average development period of the group with the single addition of BCAA (leucine, isoleucine, and valine) was similar to that of the control group (+None). This means that simply supplementing nutrients cannot compensate for the growth deficiency in a nutrient-deficient situation. The group with the single introduction of lactic acid bacteria had a shorter average development period compared to the control group and the group with the single addition of BCAA. However, it was confirmed that the growth promotion effect was statistically significantly increased in the group where lactic acid bacteria were introduced together with BCAA (see Figure 11, *** (p<0.001), ** (P<0.01), * (P<0.05), ns: not statistically significant).

[0149] 5-1-2. Effect of the combination of lactic acid bacteria and at least two types of BCAA among leucine, isoleucine, and valine Furthermore, in order to confirm whether a growth promotion effect is also exerted when leucine, isoleucine, or valine alone is used in combination with lactic acid bacteria in nutrient-deficient individuals, or when other amino acids are used in combination with lactic acid bacteria, the following experiment was conducted. While following the same experimental method as in "5-5-1." above, the types of amino acids were processed differently to observe the growth changes of nutrient-deficient Drosophila individuals.

[0150] As a result, similar to "5-5-1.", the group that only treated amino acids and did not treat lactic acid bacteria (None+3BCAA(=BCAA); None+Leucine; None+Isoleucine; None+Valine; None+Histidine; None+Leucine+Valine; None+Leucine+Isoleucine; None+Isoleucine+Valine) had an average generation period similar to that of the group that did not treat anything (None+None) (left in Figures 12A and 12B).

[0151] On the other hand, the groups that treated at least two or more of leucine, isoleucine, and valine together with lactic acid bacteria (WJL+BCAA; WJL+Leucine+Valine; WJL+Leucine+Isoleucine; WJL+Isoleucine+Valine) had a statistically significantly shortened average generation period compared to the groups treated with lactic acid bacteria or amino acids alone (right in Figure 12B). Also, the groups that treated leucine, isoleucine, or valine alone together with lactic acid bacteria (WJL+Leucine; WJL+Isoleucine; WJL+Valine) did not show a statistically significant difference in the average generation period compared to the group treated with lactic acid bacteria alone (right in Figure 12A).

[0152] From the above results, it can be confirmed that the amino acids of the present invention are not merely used in combination for nutritional supplementation, but rather play a role in enhancing the activity within the individual lactic acid bacteria by the action of two or more BCAA amino acids on the lactic acid bacteria.

[0153] 5-1-3. Effects of combinations of BCAA and various lactic acid bacteria It was confirmed whether the growth promoting effect of BCAA (leucine, isoleucine, and valine) is enhanced in nutritionally deficient individuals by other species of lactic acid bacteria other than Lactobacillus plantarum strains.

[0154] While following the same experimental method as described in the aforementioned "5-5-1.", 11 strains of Food Notice belonging to the Lactobacillus group, in which BCAA biosynthesis was previously confirmed to be deficient by comparing genetic materials (3 strains of Lactobacillus plantarum (WJL, Nizo, Nc8), 2 strains of Lactobacillus paracasei (KCTC5058, IH30-12), 3 strains of Lactobacillus rhamnosus (KCTC3237, GG, IH37-25), 2 strains of Lactobacillus fermentum (KCTC5467, IH37-57), 3 strains of Lactobacillus casei (IH37-55, IH37-56, IH37-9), 1 strain of Lactobacillus acidophilus (KCTC3594), 1 strain of Lactobacillus gasseri (KCTC3143), 2 strains of Lactobacillus bulgaricus (IH37-37, IH37-19), 1 strain of Lactobacillus reuteri (KCTC3594), 2 strains of Lactobacillus helveticus (KCTC3545, KCTC15060) and 2 strains of Lactobacillus salivarius (KCTC3157, IH37-38)) were used for the experiment.

[0155] As a result, it was confirmed that the growth promotion effect of lactic acid bacteria was significantly enhanced by the combined treatment with BCAA in other species of lactic acid bacteria other than Lactobacillus plantarum (see Figures 13 and 14).

[0156] 5-2. Confirmation of the growth promotion effect and intestinal injury improvement effect when lactic acid bacteria and BCAA are combined and treated in malnourished individuals Based on the fact that growth inhibition or intestinal injury may occur in malnourished individuals, it was confirmed whether the growth promotion and intestinal injury inhibition effects of the mouse model were shown when lactic acid bacteria and BCAA were combined and treated.

[0157] Specifically, a young mouse animal model at 3 weeks of age after weaning and separation was used as the subject, and a diet deficient in nutritional components was given. After treatment with lactic acid bacteria and BCAA (leucine, isoleucine, and valine) alone or in combination, the presence or absence of growth and improvement of intestinal injury was confirmed: control group (None); L. plantarum WJL single treatment group (+WJL); L. plantarum WJL and BCAA (leucine, isoleucine, and valine) treatment group (+WJL+BCAA). 4 to 5 mice were set in each group, the diet was supplied daily, and indicators such as body weight were tracked every two days. After 12 weeks of observation, the mice were dissected to confirm intestinal leakage (FITC), the degree of bone growth, bone mineral density, and symptom indicators of other organs. To measure intestinal leakage, the same amount of fluorescein isothiocyanate-dextran (FITC-dextran, sigma#FD4), a fluorescent substance, was given to each mouse, and the FITC fluorescence in the blood was measured 4 hours later. To measure the bone mineral density of the mice, the femurs of the mice were dissected and sampled, and then transferred to a 70% ethanol solution and stored at 4°C. In order to use a micro-CT device (SkyScan 1276, Bruker), the bone was transferred from the 70% ethanol solution to triple distilled water 48 hours before imaging and stored at 4°C. The bone images were taken using the SkyScan 1276 program. During imaging, the voxel size was fixed at 32 μm, and imaging was performed under the conditions of 70 kV and 57 μA. The NRecon program was used to reconstruct the taken images in 3D. The CTAn program was used to measure BMD (bone mineral density), bone length, and cortical bone thickness using the 3D-reconstructed images. To measure BMD, 0.25 g / cm 3 , 0.75 g / cm 3After obtaining the BMD proportional formula using two bone density phantoms with the density of , the mid-diaphysis part of the femur was specified for measurement. The entire femur was dragged and specified, and the actual bone length was measured using the calculation method inherent in the program. The cortical bone thickness was measured for the middle cross-section of the mid-diaphysis part of the femur.

[0158] As a result, in the group of mice treated with both lactic acid bacteria and BCAA (leucine, isoleucine, and valine), a statistically significant weight gain effect and intestinal leakage inhibitory effect were achieved (see Figure 15). Also, when lactic acid bacteria and BCAA (leucine, isoleucine, and valine) were introduced together, the bone density increased statistically significantly compared to when each of them was treated alone (see Figure 16). On the other hand, as shown in Figure 16, although it can be said that the bone density increased even when lactic acid bacteria were treated alone and the lactic acid bacteria themselves also had a growth promoting effect, it was confirmed that BCAA significantly increased such a growth promoting effect of lactic acid bacteria.

[0159] From the above-mentioned experiments, it was confirmed that the growth of nutritionally deficient individuals was promoted and the indicators related to diseases were improved when lactic acid bacteria alone or BCAA, especially when lactic acid bacteria were used in combination with at least two or more of leucine, isoleucine, and valine, compared to the case of treating lactic acid bacteria alone.

[0160] [Microorganism Deposit Certificate] JPEG0007697714000003.jpg185139

Claims

Claim 1: A composition for enhancing the effects of lactic acid bacteria for enhancing the effects of lactic acid bacteria, comprising leucine and valine, isoleucine and valine, or leucine, isoleucine and valine, wherein the effect of the lactic acid bacteria is to promote the growth of an individual or inhibit intestinal damage, a composition for enhancing the effects of lactic acid bacteria.

2. The composition for enhancing the effects of lactic acid bacteria according to claim 1, comprising the leucine, the isoleucine and the valine.

3. The composition for enhancing the effects of lactic acid bacteria according to claim 1, wherein promoting the growth of the individual is to increase at least one of the body weight or bone density of a nutritionally deficient individual, and inhibiting the intestinal damage is to inhibit intestinal leakage of a nutritionally deficient individual.

4. The composition for enhancing the effects of lactic acid bacteria according to claim 1, wherein the lactic acid bacteria is at least one selected from the group consisting of Lactobacillus spp., Lactococcus spp., Enterococcus spp., Streptococcus spp. and Bififobacterium spp.

5. The Lactobacillus genus is at least one selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius. The composition for enhancing the effect of lactic acid bacteria according to claim 4.

6. A pharmaceutical composition for promoting growth or inhibiting intestinal injury, comprising the composition according to any one of claims 1 to 5 and lactic acid bacteria.

7. The pharmaceutical composition according to claim 6, wherein the lactic acid bacterium is at least one selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri, Lactobacillus helveticus, and Lactobacillus salivarius.

8. A pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of growth disorder, failure to thrive, osteoporosis, osteomalacia, osteopenia, environmental enteropathy, and intestinal leakage syndrome, comprising the composition according to any one of claims 1 to 5 and a lactic acid bacterium.

9. A food composition for promoting growth or inhibiting intestinal injury, comprising the composition according to any one of claims 1 to 5 and a lactic acid bacterium.

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