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WO2025121694A1 - Composition comprenant lactobacillus plantarum hem20865, ayant une excellente activité protéolytique, ou culture, lysat ou extrait de celui-ci en tant que principe actif - Google Patents

Composition comprenant lactobacillus plantarum hem20865, ayant une excellente activité protéolytique, ou culture, lysat ou extrait de celui-ci en tant que principe actif Download PDF

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Publication number
WO2025121694A1
WO2025121694A1 PCT/KR2024/017426 KR2024017426W WO2025121694A1 WO 2025121694 A1 WO2025121694 A1 WO 2025121694A1 KR 2024017426 W KR2024017426 W KR 2024017426W WO 2025121694 A1 WO2025121694 A1 WO 2025121694A1
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WIPO (PCT)
Prior art keywords
hem20865
protein
strain
lactobacillus plantarum
culture
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Pending
Application number
PCT/KR2024/017426
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English (en)
Korean (ko)
Inventor
지요셉
정은성
서동호
심인선
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
HEM Pharma Inc
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HEM Pharma Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Priority claimed from KR1020240146706A external-priority patent/KR20250086492A/ko
Application filed by HEM Pharma Inc filed Critical HEM Pharma Inc
Publication of WO2025121694A1 publication Critical patent/WO2025121694A1/fr
Pending legal-status Critical Current
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

Definitions

  • the present invention relates to a composition
  • a composition comprising, as an active ingredient, a Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof capable of improving the protein decomposition ability of an enzyme.
  • the strain according to the present invention not only exhibits excellent protein decomposition ability alone, but also further increases protein decomposition ability when used together with an enzyme, so that it can be effectively utilized in the manufacture of general foods or health functional foods.
  • Probiotics are beneficial microorganisms that live in the intestines or other parts of humans, and are live bacteria that have a positive effect on health. They mainly include lactic acid bacteria and vitiligo, and are known to have various health-promoting effects, such as improving the intestinal environment and enhancing immunity (Ministry of Food and Drug Safety, Probiotics Safety Evaluation Guide, 2021). Probiotics sold as health functional foods must regulate intestinal flora, be non-toxic, and non-pathogenic.
  • the lactic acid bacteria that received the most approvals for individual raw material approvals by domestic lactic acid bacteria species was Lactobacillus , and among them, the plantarum species received the most approvals with 7.
  • Lactobacillus plantarum accounted for the largest number of patent applications by lactic acid bacteria, with 68 out of a total of 279.
  • Lactobacillus plantarum is establishing itself as one of the important topics in probiotics research for health promotion.
  • the object of the present invention is to provide a food composition and a health functional food comprising Lactobacillus plantarum HEM20865 strain, which has excellent protein decomposition ability and, in particular, exhibits further enhanced protein decomposition ability when combined with an enzyme.
  • the above object is only one example of the present invention, and there may be additional objects that can be achieved by the technical idea of the present invention.
  • the solution to the problem of the present invention includes the following aspects.
  • the first aspect of the present invention provides Lactobacillus plantarum HEM20865 strain ( Lactobacillus plantarum HEM20865; Accession No. KCTC 15551BP).
  • the second aspect of the present invention provides a composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof as an effective ingredient.
  • the third aspect of the present invention provides a composition comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof and a protein decomposing enzyme as active ingredients.
  • the fourth aspect of the present invention provides a food composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof as an effective ingredient.
  • the fifth aspect of the present invention provides a pharmaceutical composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof as an effective ingredient.
  • the sixth aspect of the present invention provides a method for improving protein degradation, comprising the step of administering to a subject in need thereof at least one of Lactobacillus plantarum HEM20865 strain or a culture, a lysate, or an extract thereof.
  • the seventh aspect of the present invention provides a use for improving protein decomposition ability of at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof.
  • the eighth aspect of the present invention provides the use of at least one of Lactobacillus plantarum HEM20865 strain or a culture, a lysate, or an extract thereof for producing a protein degradation enhancing agent.
  • the present invention can be usefully utilized to improve protein decomposition and digestion ability by providing probiotic Lactobacillus plantarum HEM20865, or a food composition or pharmaceutical composition containing the strain, to people who have difficulty in protein digestion.
  • Figure 1 is a drawing showing the amount of protein decomposition measured through treatment with four strains of Lactobacillus plantarum (HEM20865, HEM21080, HEM21089, HEM21113) and enzyme alone.
  • Figure 2 is a drawing showing the amount of protein decomposition measured through four strains of Lactobacillus plantarum (HEM20865, HEM21080, HEM21089, HEM21113) and enzyme combination treatment.
  • Figure 3 is a drawing comparing the amount of protein decomposition according to treatment with four strains of Lactobacillus plantarum (HEM20865, HEM21080, HEM21089, HEM21113) and enzymes alone or in combination.
  • HEM20865, HEM21080, HEM21089, HEM21113 strains of Lactobacillus plantarum
  • Figure 4 is a drawing comparing the protein decomposition multiples of skim milk, soy protein, pea protein, gluten, and total protein for each of the enzyme-only treatment group, the Lactobacillus plantarum HEM20865-only treatment group, and the Lactobacillus plantarum HEM20865 and enzyme combination treatment group compared to the control group.
  • 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.
  • references to “A and/or B” mean “A or B, or A and B.”
  • culture used throughout this specification includes a substance obtained by culturing the strain of the present invention in a known liquid medium or solid medium, and may be used interchangeably with “culture”.
  • 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.
  • health functional food used throughout this specification means a food manufactured and processed using raw materials or ingredients with functionality useful to the human body according to Act No. 6727 on Health Functional Foods, and "functionality” means obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological effects.
  • the food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process.
  • the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as food.
  • the food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long period of time because it uses food as a raw material, and is highly portable.
  • the first aspect of the present invention provides a Lactobacillus plantarum HEM20865 strain.
  • the Lactobacillus plantarum HEM20865 strain is a strain derived from Sikhye. The strain was deposited with the Korea Center for Biological Resources (KCTC) on August 14, 2023 under the deposit number KCTC 15551BP.
  • the strain according to the present invention may promote protein decomposition.
  • the second aspect of the present invention provides a composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof as an effective ingredient.
  • the third aspect of the present invention provides a composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof and a protein decomposition enzyme as active ingredients.
  • a composition for improving protein decomposition ability comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof and a protein decomposition enzyme as active ingredients.
  • the enzyme is a biocatalyst that increases the speed of metabolism by lowering the activation energy of a chemical reaction by forming an enzyme-substrate complex by binding to a substrate, and specifically, may be a protein decomposition enzyme, and in one embodiment of the present invention, a fermentation enzyme was used, but is not limited thereto.
  • the fourth aspect of the present invention provides, in one embodiment, a food composition for improving protein decomposition ability, comprising at least one of Lactobacillus plantarum HEM20865 strain or a culture, a lysate, and an extract thereof as an effective ingredient.
  • the fifth aspect of the present invention provides, in one embodiment, a pharmaceutical composition for improving protein degradation ability, comprising Lactobacillus plantarum HEM20865 strain or at least one of a culture, a lysate, and an extract thereof as an effective ingredient.
  • composition according to the second to fifth aspects of the present invention can be manufactured into a formulation that can be ingested by an individual.
  • the term 'individual' should be interpreted to include all animals that can exhibit an increased protein decomposition effect, such as humans, monkeys, pigs, cows, and chickens.
  • the term 'formulation' herein includes all that are manufactured by a method commonly used in the art, and can be in the form of, for example, powder, tablets, liquid capsules, etc.
  • the sixth aspect of the present invention provides a method for improving protein degradation, comprising the step of administering to a subject in need thereof at least one of Lactobacillus plantarum HEM20865 strain or a culture, a lysate, or an extract thereof.
  • the seventh aspect of the present invention provides a use for improving protein decomposition ability of at least one of Lactobacillus plantarum HEM20865 strain or a culture, lysate, or extract thereof.
  • the eighth aspect of the present invention provides the use of at least one of Lactobacillus plantarum HEM20865 strain or a culture, a lysate, or an extract thereof for producing a protein degradation enhancing agent.
  • Example 1 Comparison of protein decomposition ability when treated with four types of Lactobacillus plantarum and enzyme alone
  • the four strains used in this example are all Lactobacillus plantarum, including HEM20865 isolated from Sikhye, HEM21080 isolated from Chestnut, HEM21089 isolated from Lemon, and HEM21113 isolated from Tomato. All strains were cultured in MRS medium containing 10% skim milk as a cryoprotectant, and strains with a bacterial count of 1.0E+11 CFU/g or more were used in the experiment.
  • the enzyme used in this experiment was a mixture of commercially available fermented enzyme powder [fruit and vegetable mixture (resistant maltodextrin, carrot concentrate, tart cherry concentrate, Bacillus amyloliquefaciens, chicory extract powder, apple concentrate, fructooligosaccharide, red beet concentrate, white grape concentrate, mango concentrate, pomegranate concentrate, cabbage concentrate, broccoli concentrate, Jerusalem artichoke extract powder, grapefruit extract powder), organic skim milk powder, purified water, grain yogurt (brown rice, corn, domestic wheat, black rice, barley, frost laver, Job's tears, glutinous rice, sorghum, Lactobacillus plantarum, Lactobacillus casei, Pediococcus acidilactici, Bacillus), BioCore Optimum K].
  • fruit and vegetable mixture resistant maltodextrin, carrot concentrate, tart cherry concentrate, Bacillus amyloliquefaciens, chicory extract powder, apple concentrate, fructooligosaccharide, red be
  • PMAS Personalized Pharmaceutical Meta-Analysis Screening
  • PMAS Personalized Pharmaceutical Meta-Analysis Screening
  • PMAS technology which is forming an intestinal environment identically/similarly in vitro
  • four types of proteins were added to the medium respectively, and then specific strains and enzymes were treated.
  • the protein content in the medium decreased, and a strain and enzyme combination showing a high protein decomposition amount was selected.
  • Patent Registration Nos. 10-2227382 and 10-2124474 For details on the experimental method, refer to Patent Registration Nos. 10-2227382 and 10-2124474.
  • the culture time was set to 0 and 24 hours to compare before and after culture.
  • the treatment groups consisted of enzyme only, strain only, and strain and enzyme combined treatment groups.
  • the strains were treated to 9.0E+09 CFU/g in each well to ensure the same viable cell count.
  • the control group was not treated with strain or enzyme.
  • the four types of proteins were also dissolved in the medium and treated so that 2.5 mg was added to each well.
  • PMAS experiments were performed twice considering non-homogenization of proteins or enzymes, and culture was performed in an anaerobic chamber. After culturing for 0 and 24 hours, the samples were taken out from the anaerobic chamber, centrifuged at room temperature (20 degrees) for 10 minutes, and the supernatant was separated. The supernatant sample was diluted 1/10 in distilled water (Fisher) and used for protein measurement. Protein measurement was performed according to the protocol using the Quick StartTM Bradford Protein Assay (Bio-Rad) kit. After subtracting the background and average values from the measured values, the protein concentration value was determined according to the standard curve concentration. The protein concentration value was used by multiplying the dilution factor by 10 times as much as the 1/10 diluted sample was used.
  • Lactobacillus plantarum HEM20865 When four Lactobacillus plantarum strains, HEM20865, HEM21080, HEM21089, and HEM21113, were treated alone with enzymes, the degree of protein degradation in four proteins (skim milk / soybean protein / pea protein / gluten) was compared with the control group. As a result, the Lactobacillus plantarum HEM20865 strain showed a higher protein degradation amount than the other Lactobacillus plantarum strains (Fig. 1, Table 1) .
  • the protein decomposition amount was high in the following order: HEM21113, HEM20865, HEM21089, HEM21080 strains; in soybean protein, HEM20865, HEM21089, HEM21080, HEM21113 strains; in pea protein, HEM20865, HEM21089, HEM21080, HEM21113 strains; and in gluten, HEM20865, HEM21089, HEM21080, HEM21113 strains. That is, in most cases, it was confirmed that the HEM20865 strain according to the present invention had the highest degree of protein decomposition.
  • the protein degradation amount is graphed by subtracting the values measured after 0 hours of culture and 24 hours of culture.
  • the protein measurement value uses the average of two replicates.
  • Standard deviation is the standard deviation of two replicate values.
  • the degree of protein degradation in four proteins was compared with the control group by combining enzymes with four Lactobacillus plantarum strains, HEM20865, HEM201080, HEM21089, and HEM21113.
  • Lactobacillus plantarum HEM20865 strain was mixed with enzymes, a higher amount of protein degradation was observed than when other Lactobacillus plantarum strains and enzymes were mixed (Fig. 2, Table 2) .
  • the protein decomposition amount was high in the following order: HEM21113, HEM20865, HEM21080, HEM21089 strains; in soybean protein, HEM20865, HEM21089, HEM21080, HEM21113 strains; in pea protein, HEM20865, HEM21089, HEM21080, HEM21113 strains; and in gluten, HEM20865, HEM21113, HEM21080 HEM21089 strains. In most cases, the degree of protein decomposition of the HEM20865 strain claimed in the present invention was confirmed to be the highest.
  • the protein degradation amount is graphed by subtracting the values measured after 0 hours of culture and 24 hours of culture.
  • the protein measurement value uses the average of two replicates.
  • Standard deviation is the standard deviation of two replicate values.
  • the Lactobacillus plantarum HEM20865 strain showed considerably high protein decomposition ability even on its own, and showed even higher protein decomposition ability when interacting with enzymes.
  • the decomposition amount was expressed as a % based on the amount of protein detected at 0 hour of culture
  • the Lactobacillus plantarum HEM20865 strain showed a high decomposition ability of about 15% compared to other strains of the same type, although there were differences depending on the type of protein.
  • the strain and enzyme were combined, the protein decomposition ability increased by 1 to 11% compared to the single treatment (Table 3) .
  • the total protein amount which is the sum of the decomposition amounts of each protein of skim milk, soybean protein, pea protein, and gluten and the four types of proteins, was expressed as a multiple of the protein decomposition amounts of enzyme, HEM20865, and HEM20865 and enzyme mixture based on the protein amount of the control group (Fig. 4) .
  • the decomposition amount of the four types of proteins and the total protein amount was 3.0 to 3.9 times higher than that of the control group, showing the best decomposition ability.
  • the decomposition amount was 2.4 to 3.1 times higher than that of the control group, showing a higher protein decomposition ability than the enzyme group for all proteins.
  • the strain according to the present invention can promote protein decomposition alone, and can further promote protein decomposition when used together with an enzyme.
  • the amount of protein detected based on 0 hours of culture is expressed as a % of the amount of decomposition by strain alone and the amount of decomposition by strain + enzyme.
  • the HEM20865 strain showed a high decomposition ability of about 15% depending on the protein type compared to other strains of the same type, and when treated with enzymes, the protein decomposition ability increased by 1-11% compared to treatment alone.

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Abstract

La présente invention concerne une composition comprenant, en tant que principe actif, une souche de Lactobacillus plantarum HEM20865, permettant d'améliorer l'activité protéolytique d'une enzyme, ou une culture, un lysat ou un extrait de celle-ci. La présente invention concerne Lactobacillus plantarum HEM20865, qui est une souche probiotique, ou une composition alimentaire ou une composition pharmaceutique qui comprennent la souche, pour des personnes qui ont une difficulté dans la digestion des protéines. Ainsi, la souche ou les compositions peuvent être utilisées efficacement pour améliorer la dégradation des protéines et la capacité de digestion.
PCT/KR2024/017426 2023-12-06 2024-11-06 Composition comprenant lactobacillus plantarum hem20865, ayant une excellente activité protéolytique, ou culture, lysat ou extrait de celui-ci en tant que principe actif Pending WO2025121694A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2023-0175841 2023-12-06
KR20230175841 2023-12-06
KR1020240146706A KR20250086492A (ko) 2023-12-06 2024-10-24 단백질 분해능이 우수한 락토바실러스 플란타룸(Lactobacillus plantarum) HEM20865 또는 이의 배양물, 파쇄물, 추출물을 유효성분으로 포함하는 조성물
KR10-2024-0146706 2024-10-24

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KR101586778B1 (ko) * 2011-02-25 2016-02-02 오위 테르비슬리쿠 피마 바이오테흐놀로지아테 아렌두스케스쿠스 분리된 미생물 균주 락토바실러스 플란타룸 mcc1 dsm 23881 및 락토바실러스 가서리 mcc2 dsm 23882와 그의 용도
CN112063548A (zh) * 2020-08-19 2020-12-11 杭州医学院 一株具有产蛋白酶能力的植物乳杆菌及其应用
KR102273108B1 (ko) * 2019-11-21 2021-07-06 (주)서울에프엔비 프로바이오틱 활성을 갖는 신규 락토바실러스 플란타럼 kc23 균주 및 참당귀 추출물을 이용한 발효조성물
KR20230040255A (ko) * 2021-09-15 2023-03-22 일동바이오사이언스(주) 단백질 분해능을 가지는 락토바실러스 플란타럼 idcc 3501을 포함하는 식품 조성물 및 건강기능식품

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KR101586778B1 (ko) * 2011-02-25 2016-02-02 오위 테르비슬리쿠 피마 바이오테흐놀로지아테 아렌두스케스쿠스 분리된 미생물 균주 락토바실러스 플란타룸 mcc1 dsm 23881 및 락토바실러스 가서리 mcc2 dsm 23882와 그의 용도
KR102273108B1 (ko) * 2019-11-21 2021-07-06 (주)서울에프엔비 프로바이오틱 활성을 갖는 신규 락토바실러스 플란타럼 kc23 균주 및 참당귀 추출물을 이용한 발효조성물
CN112063548A (zh) * 2020-08-19 2020-12-11 杭州医学院 一株具有产蛋白酶能力的植物乳杆菌及其应用
KR20230040255A (ko) * 2021-09-15 2023-03-22 일동바이오사이언스(주) 단백질 분해능을 가지는 락토바실러스 플란타럼 idcc 3501을 포함하는 식품 조성물 및 건강기능식품

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Title
DUAN, C. ET AL.: "Proteolytic activity of Lactobacillus plantarum strains in cheddar cheese as adjunct culture s", JOURNAL OF FOOD PROTECTION, vol. 82, no. 12, 2019, pages 2108 - 2118, XP093290839, DOI: 10.4315/0362-028X.JFP-19-276 *

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