Composition comprising kimchi-derived lactiplantibacillus plantarum LM1001 strain having excellent proteolytic activity as active ingredient

Abstract

The present invention relates to a Lactiplantibacillus plantarum) LM1001 (Accession No. KCCM13370P) strain, and a composition for promoting proteolysis, comprising, as an active ingredient, one or more of a culture, lysate, or extract of the strain. The Lactiplantibacillus plantarum LM1001 strain according to the present invention has excellent proteolytic activity, can produce branched-chain amino acids (glutamic acid, tyrosine, alanine, and lysine) that can help produce muscle and prevent muscle loss by degrading whey protein, has excellent β-galactosidase or α-galactosidase activity, and does not have β-glucuronidase activity, and thus the present invention has the effect of ameliorating indigestion, diarrhea, abdominal bloating, and the like due to protein intake.

Description

A composition comprising kimchi-derived Lactobacillus plantarum LM1001 strain with excellent protein decomposition ability as an active ingredient.

[0001] The present invention relates to a composition comprising, as an active ingredient, Lactobacillus plantarum LM1001, a lactic acid bacterium derived from kimchi, which can produce protein-decomposing enzymes to decompose dietary proteins, produce amino acids that can promote muscle growth, and improve lactose intolerance.

[0002] The rise in obesity due to age-related muscle loss, Westernized eating habits, and reduced activity levels is a significant public health issue. Protein helps maintain muscle mass while resisting age-related muscle loss. Adequate protein intake is known to reduce the risk of sarcopenia and obesity-related diseases across all age groups.

[0003] Dietary protein sources can be divided into animal proteins, such as milk, eggs, beef, pork, chicken, and fish, and plant proteins, such as soybeans and grains. Among these, milk protein or whey protein, unlike plant proteins, boasts a superior composition of essential amino acids. In particular, it contains a high concentration of branched-chain amino acids (BCAAs), which are beneficial for muscle growth. This helps with muscle biosynthesis, preventing muscle loss, and enhancing immune response.

[0004] Since proteins are high molecular weight substances, they must be broken down in order to be absorbed into the body. In the body, protein-digesting enzymes such as pepsin in the stomach and trypsin and chymotrypsin in the small intestine are produced to break down the peptide chains of proteins and ultimately be absorbed from the villi of the small intestine in the form of amino acids.

[0005] Protein breakdown involves a variety of enzymes in various digestive organs, and gut microbes are known to influence protein breakdown and absorption. Proteins, being large molecules, can cause indigestion depending on an individual's digestive health. The gas produced during the breakdown process can also cause bloating, abdominal distension, flatulence, and increased odor from stool.

[0006] In particular, whey protein concentrate, which is widely used as a protein supplement, contains about 5% lactose. If you have lactose intolerance due to a lack of lactase, gastrointestinal problems such as abdominal pain, diarrhea, and vomiting may occur after consuming whey protein concentrate because the lactose is not broken down in the digestive system.

[0007] Lactose intolerance can occur due to disease or injury to the small intestine, or after small intestine resection, but most often it is caused by a congenital deficiency of the enzyme lactase. While lactose intolerance is very common worldwide, affecting approximately 67% of the population, there are significant racial differences. While it is estimated to be around 10% in Europe, it is estimated to be around 75% in Korea. Furthermore, the incidence of lactose intolerance increases with age, with approximately 25% of school-age Koreans and 75% of adults suffering from it. Furthermore, lactose intolerance differs by gender, with a higher incidence in males (80%) and females (73%) in Korean adults.

[0008] Therefore, probiotics that can promote the digestion of whey protein concentrate, which is widely used as dietary protein in the digestive system, and can break down lactose are expected to have great industrial value.

[0009] Amino acids are the building blocks of protein. Dietary protein is broken down into various amino acids after ingestion, which are then digested and absorbed. In the human body, amino acids promote muscle formation, structure the body, regulate essential functions, and provide energy.

[0010] Among amino acids, branched-chain amino acids (BCAs) can help improve exercise performance by promoting muscle production, growth, and regeneration. They are also known to be effective in weight loss and reducing post-exercise fatigue. Valine, isothiocyanate, and leucine are examples of these amino acids. Leucine, in particular, plays a signaling role in muscle biosynthesis pathways and is known to help alleviate sarcopenia in the elderly.

[0011] In addition to branched-chain amino acids, certain amino acids are known to increase muscle building and exercise performance, such as glutamic acid, a precursor to glutamine that plays a role in protein synthesis and muscle recovery; lysine, which can help with skeletal muscle growth and protein breakdown inhibition; tyrosine, which can help improve exercise performance; and alanine, which increases the bioavailability of branched-chain amino acids in the body.

[0012] Recently, as the health-promoting effects of protein have become widely known, protein intake has increased. Research is also growing on how to increase muscle mass and prevent muscle loss through the proper breakdown of ingested protein, which leads to the production and absorption of essential amino acids. Therefore, research on probiotics, which are taken with protein to convert dietary protein into health-promoting amino acids and increase its absorption rate, is gaining increasing importance.

[0013] Gut microbes can contribute to the digestion and absorption of food and can also influence the breakdown of dietary proteins. Therefore, the present project aims to develop a probiotic composition that can promote protein breakdown or increase branched-chain amino acids and other muscle-building amino acids when taken with dietary protein, thereby increasing muscle biosynthesis.

[0014] The present invention provides a composition for promoting protein decomposition, which comprises at least one of the Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P) strain, a culture, a lysate, and an extract of the strain as an active ingredient.

[0015] The strain may have whey protein decomposition ability, may have the ability to produce at least one of valine, isoleucine, leucine, glutamic acid, tyrosine, alanine, and lysine, may have β-galactosidase or α-galactosidase activity, and may not have β-glucuronidase activity.

[0016] In addition, the present invention provides a food composition for promoting protein decomposition, a pharmaceutical composition for preventing or treating lactose intolerance, and a health functional food composition for promoting protein decomposition, which contain at least one of the strain, a culture, a fragment, and an extract of the strain as an active ingredient.

[0017] The Lactiplantibacillus plantarum LM1001 strain according to the present invention has excellent protein decomposition ability, can decompose whey protein to produce branched-chain amino acids (glutamic acid, tyrosine, alanine, lysine) that can help build muscle and prevent muscle loss, and has excellent β-galactosidase or α-galactosidase activity and no β-glucuronidase activity, so the present invention is effective in improving indigestion, diarrhea, abdominal distension, etc. caused by protein intake.

[0018] Figure 1 is a drawing showing the results of comparative measurements of valine, isoleucine, leucine, and the total amount thereof of the strain in an experiment to determine the branched-chain amino acid production ability of the Lactibacillus plantarum LM1001 strain.

[0019] Figure 2 is a drawing showing the results of comparative measurement of the amounts of glutamic acid, tyrosine, alanine, and lysine in the Lactibacillus plantarum LM1001 strain.

[0020] Below, with reference to the attached drawings, embodiments of the present invention are described in detail to facilitate easy implementation by those skilled in the art. However, the present invention can be implemented in various different forms and is not limited to the embodiments described herein. In the drawings, irrelevant parts have been omitted for clarity, and similar reference numerals have been used throughout the specification to indicate similar elements.

[0021] Throughout this specification, whenever a part is said to "include" a component, this means that it may include other components, but not to the exclusion of other components, unless otherwise stated.

[0022] The terms “step of” or “step of” used throughout this specification do not mean “step for”.

[0023] Throughout this specification, the term "combination(s) thereof" included in the expressions in the Makushi format means one or more mixtures or combinations selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

[0024] Throughout this specification, references to “A and / or B” mean “A or B, or A and B.”

[0025] This document explains the amino acids mentioned throughout this specification. Amino acids are the building blocks of proteins. After ingestion, dietary protein is broken down into various amino acids, which are then digested and absorbed. In the human body, amino acids promote muscle formation, form the body, regulate essential functions, and provide energy.

[0026] Branched-chain amino acids (BCAAs) can help improve exercise performance by promoting muscle production, growth, and regeneration. They are also effective in weight loss and reducing post-exercise fatigue. These amino acids include valine, isoleucine, and leucine. Leucine, in particular, plays a signaling role in muscle biosynthesis pathways and is known to help improve sarcopenia in the elderly.

[0027] In addition to branched-chain amino acids, certain amino acids are known to increase muscle building and exercise performance, such as phenylalanine, which can help stimulate protein synthesis and alleviate muscle loss; lysine, which can help skeletal muscle growth and inhibit protein breakdown; threonine, which acts as a protein synthesis precursor and promotes muscle protein synthesis; and alanine, which increases bioavailability by inhibiting the breakdown of branched-chain amino acids in the body.

[0028] Additionally, certain amino acids are known to increase muscle building and exercise performance, including glutamic acid, a precursor to glutamine, which plays a role in protein synthesis and muscle recovery; lysine, which can help with skeletal muscle growth and inhibition of protein breakdown; tyrosine, which can help improve exercise performance; and alanine, which increases bioavailability by inhibiting the breakdown of branched-chain amino acids in the body.

[0029] The present invention provides a composition comprising at least one of the strain Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P), a culture, a lysate, and an extract of the strain as an active ingredient, the composition comprising a composition for promoting protein decomposition, a food composition for promoting protein decomposition, a pharmaceutical composition for preventing or treating lactose intolerance, and a health functional food composition for promoting protein decomposition.

[0030] In one embodiment of the present invention, the Lactiplanta plantarum LM1001 strain may have a whey protein decomposition ability and may have a production ability for at least one of valine, isoleucine, leucine, glutamic acid, tyrosine, alanine, and lysine.

[0031] Additionally, the Lactiplanta LM1001 strain may have β-galactosidase or α-galactosidase activity, and may not have β-glucuronidase activity.

[0032] The term "food" as used throughout this specification includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

[0033] The food of the present invention can be manufactured by methods commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. In addition, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulation, and unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material, and is highly portable, so the food of the present invention can be consumed as a supplement to enhance protein decomposition ability and the effect of improving lactose intolerance.

[0034] The food composition of this hospital can be consumed on a daily basis, so it can be expected to be highly effective in promoting protein decomposition and improving lactose intolerance, and thus can be used very usefully.

[0035] The above food composition may additionally include a physiologically acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0036] In addition, the food composition may include additional ingredients commonly used in food compositions to improve odor, taste, sight, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), chromium (Cr), etc. In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine.

[0037] In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), sterilizers (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar colorants, etc.), color developers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (glucose fillers), film-forming agents, gum-forming agents, foam suppressants, solvents, and improvers. The above additives can be selected according to the type of food and used in an appropriate amount.

[0038] As an example, the food composition of the present invention can be used as a health beverage composition, and in this case, it can contain various flavoring agents or natural carbohydrates as additional ingredients like regular beverages. The natural carbohydrates mentioned above can be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener can be a natural sweetener such as thaumatin and stevia extract; or a synthetic sweetener such as saccharin and aspartame.

[0039] In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. Furthermore, the composition may contain fruit pulp for the production of natural fruit juice, fruit juice beverages, or vegetable beverages. These ingredients may be used independently or in combination.

[0040] In one embodiment of the present invention, the pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions, according to conventional methods, but may not be limited thereto.

[0041] In one embodiment of the present invention, when formulating the pharmaceutical composition, it may be prepared using a diluent or excipient such as a generally used filler, bulking agent, binder, wetting agent, disintegrant, or surfactant, but may not be limited thereto.

[0042] In one embodiment of the present invention, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations may be prepared by mixing the strain-derived component with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition, for example, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used, but may not be limited thereto.

[0043] In one embodiment of the present invention, liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included, but may not be limited thereto.

[0044] In one embodiment of the present invention, formulations for parenteral administration may include, but are not limited to, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. For example, non-aqueous solvents or suspensions may include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. For example, suppositories may include, but are not limited to, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin.

[0045] A pharmaceutical composition according to one embodiment of the present invention may be a pharmaceutical composition or a quasi-drug composition.

[0046] The term "quasi-drug" used throughout this specification refers to products that are used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases in humans or animals, and have a milder effect than drugs. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products other than those used for the purpose of drugs, and include products used for the treatment or prevention of diseases in humans or animals, and products that have a mild effect on the human body or do not directly affect it.

[0047] The above-mentioned pharmaceutical composition of the present invention can be manufactured in a formulation selected from the group consisting of body cleanser, disinfectant, detergent, kitchen detergent, cleaning detergent, toothpaste, mouthwash, wet tissue, detergent, soap, hand wash, hair cleanser, hair softener, humidifier filler, mask, ointment, and filter filler, but is not limited thereto.

[0048] In one embodiment of the present invention, the pharmaceutical composition may be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" herein means an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention, and the effective dosage level may be determined according to factors including the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, drugs used in combination or simultaneously with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a component known to exhibit a therapeutic effect on a known intestinal disease. It is important to take all of the above factors into consideration and administer an amount that can achieve the maximum effect with the minimum amount without side effects.

[0049] In one embodiment of the present invention, the dosage of the pharmaceutical composition can be determined by a person skilled in the art in consideration of the purpose of use, the degree of toxicity of the disease, the patient's age, weight, sex, medical history, or the type of substance used as an active ingredient.

[0050] The pharmaceutical composition of the present invention is not particularly limited thereto, but may be administered via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and rectal administration, depending on the intended purpose. However, when administered orally, it may be administered in an unformulated form, and since the strain may be denatured or decomposed by gastric acid, the oral composition may be administered orally in a form that coats the active agent or is formulated to protect it from decomposition in the stomach, or in the form of an oral patch. In addition, the composition may be administered by any device that allows the active agent to move to the target cell.

[0051] The term "health functional food" used throughout this specification refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body as defined in Act No. 6727 on Health Functional Foods, and "functionality" means obtaining a beneficial effect for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body.

[0052] The term "health functional food" refers to a food that has a more active health maintenance or promotion effect than regular foods, while "health supplement food" refers to a food intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement food" may be used interchangeably.

[0053] In the present example, the experiment was conducted mainly using a culture solution of the Lactiplantibacillus plantarum LM1001 strain. However, since probiotics are strains that exist in the intestines and produce lactic acid, etc., and have a beneficial effect on the intestinal environment, the strain itself, strain lysate, and strain extract also have the same effect as the culture solution.

[0054] In one embodiment, the present invention provides a method for promoting protein breakdown and preventing or treating lactose intolerance. The present invention also provides a composition for use in improving lactose intolerance.

[0055] In one embodiment, the present invention provides a method for promoting protein degradation, comprising administering to a subject in need thereof one or more of the strain Lactiplantibacillus plantarum LM1001 (Accession No. KCCM13370P), a culture, a lysate, or an extract of the strain. Here, the term "subject" should be interpreted to include all animals that can benefit from the promotion of protein degradation or the prevention or treatment of lactose intolerance, such as humans, monkeys, pigs, cows, and chickens.

[0056] Hereinafter, implementation examples and embodiments of the present invention will be described in detail with reference to the attached drawings. However, the present invention may not be limited to these implementation examples and embodiments and drawings.

[0057] Example 1. Selection of a strain with excellent protein decomposition ability

[0058] To evaluate the skim milk protein decomposition ability of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P), a plate medium was prepared using skim milk powder and agar, and the skim milk protein decomposition ability of the strain was confirmed along with that of comparative strains. 25 g of skim milk powder and 15 g of agar were dissolved in 250 mL of distilled water and 500 mL of distilled water, respectively, and sterilized. The two solutions were mixed and dispensed into each petri dish in 20 mL portions. After cooling to room temperature, the agar was confirmed to have solidified before use.

[0059] Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) and other comparative strains were used for the degradation evaluation after being cultured three times at 12-hour intervals. The inoculation concentration for each culture was 0.1% v / v, and the culture temperature was adjusted to 37±3℃. 4 μL of each strain's culture solution was applied to agar medium made of skim milk powder and left at room temperature for 30 minutes to allow the culture solution to sufficiently permeate the medium. After culturing in an incubator at 37℃ for 24 hours, the formation of a clear zone and the area and transparency of the clear zone were measured using the Image J program. The area of ​​the clear zone was measured as the area of ​​the transparent part due to protein degradation, and was expressed as mm 2 The transparency was converted to gray value of the transparent ring and the gray value of the medium, and the protein degradation index was calculated using the formula below. All tests were performed three times, and the average of each result was calculated and compared.

[0060] Protein degradation index = (A×B) / 100

[0061] A: Area of ​​transparent ring (mm) 2 )

[0062] B: Transparency of the transparent ring (%) = 100-(grayness of the transparent ring / grayness of the badge) x 100

[0063] As a result, it was confirmed that not all strains could decompose skim milk protein (if unable to decompose, it was marked with -). Among the experimental strains, the skim milk protein decomposition index of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) was confirmed to be superior to lactic acid bacteria belonging to the genera Bifidobacterium, Lacticazeibacillus, Lactobacillus, and Streptococcus, and it also showed a superior decomposition index among the same genus Lactiplantibacillus plantarum (Table 1).

[0064] Skim milk protein degradation index of Lactibacillus plantarum strains. Area of ​​transparent ring. Transparency of transparent ring. Protein degradation index. Agar medium---Bifidobacterium animalis lactis LP017---Bifidobacterium bifidum LP1108---Bifidobacterium longum LP102425.16.21.6LacticaeBacillus rhamnosus LP1012---Lactibacillus plantarum LM100134.110.73.6Lactibacillus plantarum LP1202---Lactibacillus plantarum LP1203---Lactibacillus plantarum LP1204---Lactibacillus plantarum LP1205---Lactibacillus plantarum LP1206---Lactibacillus plantarum LP1209---Lactibacillus plantarum LP1210---Lactibacillus plantarum LP1211---Lactobacillus acidophilus LP106073.67.73.5 Streptococcus thermophilus LP1012---

[0065] Example 2. Evaluation of whey protein decomposition ability

[0066] To evaluate the whey protein decomposition ability of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P), culture solutions were prepared using whey protein concentrate (WPC) and whey protein isolate (WPI), and the whey protein decomposition ability of the strain was evaluated along with that of comparative strains. For the WPC culture solution, 25 g of WPC powder was dissolved in 1 L of distilled water, 10 mL was dispensed into each 15 mL conical tube, and sterilized at 65°C for 30 minutes. After cooling to room temperature, the strain was used. The whey protein isolate culture solution was prepared by dissolving 10 g of whey protein isolate powder and 10 g of glucose in 1 L of distilled water, dispensing 10 mL each into 15 mL conical tubes, sterilizing at 65°C for 30 minutes, and cooling to room temperature before use.

[0067] Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) and other comparative strains were cultured three times at 12-hour intervals in the same manner as in Example 1 and then used for whey protein degradation evaluation. The culture solution of each strain was centrifuged (10,000 rpm, 10 minutes) to recover the cells, washed twice with phosphate buffered saline solution, and then used as a sample using phosphate buffered saline solution to make it 8 log CFU / mL.

[0068] After inoculating 100 μL of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) and other comparative strain samples into 10 mL of concentrated whey protein culture medium and isolated whey protein, the protein was quantified and cultured for 72 hours in an incubator at 37±3°C. The protein amount was quantified and the protein amount before and after culture was compared to evaluate the decomposition ability of each whey protein.

[0069] Protein quantification was performed using a BCA assay kit. 1 mg of bovine serum albumin was dissolved in 1 mL of distilled water to prepare a standard, which was then diluted with distilled water to 0.5 mg / mL, 0.25 mg / mL, and 0.125 mg / mL and used together with distilled water as a standard for each concentration. 50 μL of culture medium samples were collected and diluted (20-fold) in 950 μL of distilled water for use. 25 μL of the standard substances and diluted culture medium samples at concentrations of 0, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, and 1 mg / mL were dispensed into a 96-well plate, and 200 μL of BCA reagent were each treated. After incubation at 37±3°C for 30 minutes, the absorbance was measured at 562 nm. A standard curve was created using the absorbance results for each concentration of the standard substance, and the absorbance values ​​of each culture medium were input into the standard curve to measure the protein amount. Protein degradation capacity was expressed as the value obtained by subtracting the protein quantification value after 72 hours of incubation from the protein quantification value before incubation and multiplying the result by 100.

[0070] Protein decomposition (%) = (AB)x100

[0071] A: Protein quantitative value before culture

[0072] B: Protein quantification value after 72 hours of culture

[0073] As a result of comparing the concentrated whey protein decomposition ability and isolated whey protein decomposition ability of various strains under the same conditions, the types of proteins that could be decomposed were different for each strain, and the concentrated whey protein and isolated whey protein decomposition ability of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) was confirmed to be more than twice as excellent as that of comparative strains in other genera. In addition, the Lactiplantibacillus plantarum LM1001 strain was confirmed to have the best concentrated whey protein decomposition ability among the Lactiplantibacillus plantarum strains in the same genus (Table 2).

[0074] Whey protein decomposition ability of Lactibacillus plantarum strains Classification Whey concentrate protein decomposition ability (%) Whey isolate protein decomposition ability (%) Lactibacillus plantarum LM100121.15.4 Lactibacillus plantarum LP12020.011.4 Lactibacillus plantarum LP12034.68.9 Lactibacillus plantarum LP120414.02.5 Lactibacillus plantarum LP12056.97.8 Lactibacillus plantarum LP120617.293.85 Lactibacillus plantarum LP12093.46.9 Lactibacillus plantarum LP12100.61.3 Lactobacillus plantarum LP12118.711.3 Rimosylactobacillus fermentum LP11491.22.9 Bifidobacterium bifidum LP11080.06.3 Pediococcus acidilactici LP10138.11.9

[0075] Example 3. Effect of enhancing the production of amino acids that help build muscle after decomposition of concentrated whey protein.

[0076] To verify and compare the activity of Lactiplantibacillus plantarum LM1001 strain (KCCM13370P) and other Lactiplantibacillus plantarum strains in promoting the production of amino acids that can promote muscle production after whey protein concentrate decomposition, the changes in the contents of branched-chain amino acids (valine, isoleucine, leucine), glutamic acid, tyrosine, alanine, and lysine were analyzed using a high-performance liquid chromatography / photodiode array detector system.

[0077] Using concentrated whey protein as a control, the concentrated whey protein aqueous solution prepared under the same conditions as Example 2 was inoculated with the Lactiplantibacillus plantarum strain and cultured at 37°C. The amino acids in the culture solution were analyzed to measure changes in amino acids that contribute to the promotion of muscle production by decomposing the concentrated whey protein.

[0078] The 72-hour culture medium of Example 2 was centrifuged (4000 rpm, 15 minutes) and the supernatant was used as the analysis sample. The analysis sample was concentrated, dissolved in 1 ml of 0.1 N hydrochloric acid aqueous solution, and homogenized by vortex stirring. Then, free amino acids in the concentrated sample were extracted for 15 minutes in an ultrasonic bath. The free amino acid extract was centrifuged, and the supernatant was filtered. The free amino acids were analyzed using a high-performance liquid chromatograph / photodiode array detector system. The analysis method was performed using a 0.1% formic acid aqueous solution and a 0.1% formic acid acetonitrile solution as the mobile phases and an appropriate gradient elution, and the column was analyzed using Agilent's Zorbax Eclipse AAA (4.6 mm ID * 150 mm, 3.5 ㎛).

[0079] As a result, trace amounts of branched-chain amino acids were detected in concentrated whey protein, but when concentrated whey protein and Lactobacillus plantarum strains were reacted together, large amounts of valine, isoleucine, leucine, glutamic acid, tyrosine, alanine, and lysine were detected in all strains. In particular, when the Lactobacillus plantarum LM1001 strain was treated, the production of branched-chain amino acids was detected to be 2 to 2.7 times higher than that of other Lactobacillus plantarum strains, and the contents of glutamic acid, tyrosine, alanine, and lysine were also detected to be higher than those of other Lactobacillus plantarum strains (Fig. 1, Fig. 2, Table 3, Table 4).

[0080] Branched-chain amino acid, glutamic acid, tyrosine, alanine, and lysine production capacity of Lactibacillus plantarum strains (μg / mL) Branched-chain amino acid Valine Isoleucine Leucine Total Control 5.29 2.44 5.53 13.27 Lactibacillus plantarum ATCC 14197 37.1845.91 71.25 154.34 Lactibacillus plantarum LM1001 136.5893.95 173.82 404.35 Lactibacillus plantarum LP1202 36.1945.93 62.96 145.09 Lactibacillus plantarum Lactibacillus plantarum LP120335.4546.3969.18151.02 Lactibacillus plantarum LP120437.3345.7069.03 152.06 Lactibacillus plantarum LP120536.4849.0172.6 1158.10 Lactibacillus plantarum LP120644.7457.1881.54 183.46 Lactibacillus plantarum LP120928.4530.83 116.12175.40 Lactibacillus plantarum LP121030.1935.26 137.94 203.39 Lactibacillus plantarum LP121155.4133.2091.91180.52

[0081] Control group: Whey protein concentrate Comparison group: No additives

[0082] Test group: Whey protein concentrate reacted with Lactobacillus plantarum LM1001

[0083] Comparison of glutamic acid, tyrosine, alanine, and lysine production capacity of Lactibacillus plantarum strains. Measurement value (μg / mL) Amino acid Glutamic acid Tyrosine Alanine Lysine Control group 0.79 1.29 1.00 6.21 Lactibacillus plantarum ATCC 1419759.01 27.65 38.86 42.33 Lactibacillus plantarum LM 1001 142.37 65.28 66 0.02 211.95 Lactibacillus plantarum LP 1202 45.32 19.39 35.56 37.30 Lactibacillus plantarum LP 1203 53.05 23.40 37.97 36.81 Lactibacillus plantarum Lactibacillus plantarum LP120456.8625.2842.4861.53 Lactibacillus plantarum LP120556.2122.1241.8644.88 Lactibacillus plantarum LP120673.4324.8746.1953.70 Lactibacillus plantarum LP120956.1625.5523.4140.06 Lactibacillus plantarum LP121041.4613.6119.1127.12 Lactibacillus plantarum LP121167.351.80255.57137.99

[0084] Control group: Whey protein concentrate Comparison group: No additives

[0085] Test group: Whey protein concentrate reacted with Lactobacillus plantarum LM1001

[0086] Example 4. Evaluation of enzyme activity of Lactibacillus plantarum composition

[0087] The API ZYM kit was used to evaluate the enzyme activity of Lactiplantibacillus plantarum strains, including Lactiplantibacillus plantarum LM1001 strain (KCCM13370P). The cells of Lactiplantibacillus plantarum strains cultured three times in the same manner as in Example 1 were harvested and prepared using phosphate-buffered saline solution (9.18 log CFU / mL) according to the instructions for use of the API ZM Kit to a turbidity of 5 on MacFarland's scale. After that, 65 μL of the sample was treated on the strip in the kit, and the sample was incubated in an incubator at 37°C for 2 hours. The activity of each enzyme was confirmed through a color reaction by treating with the ZYM reagent. The degree of change in the color reaction was expressed as a value from 0 to 5, and only a value of 3 or more (positive reaction) was indicated with "●".

[0088] The evaluation results confirmed that enzyme activities differed by strain even within the same Lactibacillus plantarum. Among a total of 11 strains, only the Lactibacillus plantarum LM1001 strain showed the activity of beta-galactosidase, a lactose-decomposing enzyme, and alpha-galactosidase, an enzyme involved in preventing intestinal gas formation.

[0089] In addition, it was confirmed to be a safe strain as it did not show activity against β-glucuronidase, an enzyme involved in causing colon cancer and various diseases due to damage to the intestinal mucosa by producing amines and toxic substances during the metabolic process, and did not show activity against mannosidase and flucosidase, which are known to inhibit the formation of carbohydrates and have a negative effect on energy intake.

[0090] As a result of the above, it was confirmed that the composition of the invention can improve the disadvantages of consuming dairy protein without negatively affecting energy metabolism (Table 5).

[0091] Enzyme number 1: Alkaline phosphate

[0092] Enzyme number 2: Esterase (C4)

[0093] Enzyme number 3: Esterase Lipase (C8)

[0094] Enzyme number 4: Lipase (C14)

[0095] Enzyme number 5: Leucine arylamidase

[0096] Enzyme number 6: Valine arylamidase

[0097] Enzyme number 7: Crystine arylamidase

[0098] Enzyme number 8: Trypsin

[0099] Enzyme number 9: α-chymotrypsin

[0100] Enzyme number 10: Acid phosphate

[0101] Enzyme number 11: Naphtol-AS-BI-Phosphohydrolase

[0102] Enzyme number 12: α-galactosidase

[0103] Enzyme number 13: β-galactosidase

[0104] Enzyme number 14: β-glucuronidase

[0105] Enzyme number 15: α-glucosidase

[0106] Enzyme number 16: β-glucosidase

[0107] Enzyme number 17: N-acetyl-β-glucosaminidase

[0108] Enzyme number 18: α-mannosidase

[0109] Enzyme number 19: α-flucosidase

[0110] Enzyme activity evaluation of Lactibacillus plantarum strains. Classification. Enzyme number. 12345678910111213141516171819. Lactibacillus plantarum LM1001. Lactibacillus plantarum LP1202. Lactibacillus plantarum LP1203. Lactibacillus plantarum LP1204. Lactibacillus plantarum LP1205. Lactibacillus plantarum LP1206. Lactibacillus plantarum LP1209. Lactibacillus Plantarum LP1210●●●●●●Lactiplantibacillus Plantarum LP1211●●●●●

[0111] [Correction pursuant to Rule 91, September 13, 2024]

Claims

A composition for promoting protein decomposition comprising at least one of the strain Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P), a culture, a lysate, and an extract of the strain as an active ingredient. In the first paragraph, A composition for promoting protein degradation, wherein the strain has a whey protein degradation ability. In the first paragraph, A composition for promoting protein decomposition, wherein the strain has the ability to produce at least one of valine, isoleucine, leucine, glutamic acid, tyrosine, alanine, and lysine. In the first paragraph, A composition for promoting protein degradation, wherein the strain has beta-galactosidase or alpha-galactosidase activity. In paragraph 4, A composition for promoting protein degradation, wherein the strain does not have activity against β-glucuronidase. A food composition for promoting protein decomposition, comprising at least one of Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P) strain, a culture, a lysate, and an extract of the strain as an active ingredient. A pharmaceutical composition for preventing or treating lactose intolerance, comprising at least one of the strain Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P), a culture, a lysate, and an extract of the strain as an active ingredient. A health functional food composition for promoting protein decomposition, comprising at least one of Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P) strain, a culture, a lysate, and an extract of the strain as an active ingredient. A method for promoting protein degradation, comprising administering to a subject in need thereof at least one of a Lactiplantibacillus plantarum LM1001 (accession number KCCM13370P) strain, a culture, a lysate, and an extract of the strain.

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