WO2020130471A1 - Nouveau lactobacillus ayant un effet de réduction du poids corporel ou de la graisse corporelle et son utilisation - Google Patents

Nouveau lactobacillus ayant un effet de réduction du poids corporel ou de la graisse corporelle et son utilisation Download PDF

Info

Publication number
WO2020130471A1
WO2020130471A1 PCT/KR2019/017441 KR2019017441W WO2020130471A1 WO 2020130471 A1 WO2020130471 A1 WO 2020130471A1 KR 2019017441 W KR2019017441 W KR 2019017441W WO 2020130471 A1 WO2020130471 A1 WO 2020130471A1
Authority
WO
WIPO (PCT)
Prior art keywords
lactobacillus sakei
activity
accession
kccm12296p
strain
Prior art date
Application number
PCT/KR2019/017441
Other languages
English (en)
Korean (ko)
Inventor
나규흠
장환봉
오성준
한명주
김동현
Original Assignee
동화약품주식회사
경희대학교 산학협력단
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.)
Filing date
Publication date
Application filed by 동화약품주식회사, 경희대학교 산학협력단 filed Critical 동화약품주식회사
Publication of WO2020130471A1 publication Critical patent/WO2020130471A1/fr

Links

Images

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • 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
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/179—Sakei
    • 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

Definitions

  • the present invention relates to a novel lactic acid bacteria, more specifically, isolated from kimchi, obesity suppression effect, body fat reduction effect, blood sugar control effect, blood cholesterol lowering effect, blood neutral lipid lowering effect, endotoxin lowering effect, atherosclerosis inhibiting effect , It relates to a novel lactic acid bacteria having various functionalities such as inflammation inhibitory effect.
  • the present invention relates to various uses of new lactic acid bacteria, and more specifically, prevention of inflammatory diseases including obesity, fatty liver, diabetes, dyslipidemia, endotoxemia, metabolic syndrome, lipid-related metabolic disease or colitis of new lactic acid bacteria. , Improvement or treatment use.
  • lifestyle-related diseases meaning diseases caused by lifestyle changes.
  • Obesity, hyperlipidemia, diabetes, and hypertension among lifestyle-related diseases are known to be important risk factors for cardiovascular disease, and a case in which these cardiovascular risk factors are simultaneously multiple is called metabolic syndrome.
  • metabolic syndrome refers to a condition in which insulin resistance is severe, and the risk of diabetes and cardiovascular disease is very high.
  • the risk of developing cardiovascular disease is more than doubled, and the probability of developing diabetes is increased by 10 times or more. It is known.
  • the prevalence of various chronic diseases such as arthritis and cancer is increasing due to an increase in lifestyle-related diseases.
  • Obesity refers to a state in which energy is excessively accumulated in the body due to an imbalance in energy consumption and consumption.
  • obesity is defined as obesity when it exceeds 20% of the standard weight.
  • Obesity is an accelerator that causes adult disease and increases the risk of developing type 2 diabetes and arteriosclerosis as a metabolic disorder, and burdens the heart's blood supply, resulting in heart disease. , Liver cirrhosis.
  • the weight of the body burdens the bones and joints, skeletal abnormalities occur and activity is limited as the behavior is slowed, the lack of exercise leads to further obesity, and in particular, female obesity causes endocrine abnormalities, resulting in menstrual disorders, decreased libido, and childbirth. Complications, eczema, sweat a lot. As such, the incidence of hyperlipidemia, hypertension, arteriosclerosis, diabetes, fatty liver, and joint abnormalities caused by excessive fat due to obesity is increasing significantly.
  • Korean Patent Publication No. 10-1108428 discloses a pharmaceutical composition for preventing and treating obesity using Lactobacillus sp. BNR17.
  • Korean Patent Publication No. 10-0686557 discloses a pharmaceutical composition for preventing and treating obesity using Lactobacillus sp. PL60.
  • Republic of Korea Patent Publication No. 10-1061219 discloses a pharmaceutical composition for preventing and treating diabetes, including as an active ingredient, a pig potato fermented extract fermented with Lactobacillus sp .
  • Korean Patent Publication No. 10-1407980 discloses the effect of reducing the levels of insulin, risistin, glucose, C-peptide, and triglyceride in the blood.
  • Hyperinsulinemia hyperglycemia , containing Lactobacillus curvatus HY7601 (Accession No.: KCTC 11456BP) and Lactobacillus plantarum KY1032 (Accession No.: KCCM10430) as active ingredients. And health triglycerides for improving hypertriglyceridemia.
  • Republic of Korea Patent Publication No. 10-1708173, Lactobacillus sakei (Lactobacillus sakei) OK67 accesion No.: KCCM11670P
  • its culture, its fragments or extracts thereof as an active ingredient, metabolic syndrome, prevention of inflammatory diseases Or therapeutic compositions are disclosed.
  • the present invention has been derived under such a conventional background, the object of the present invention is to suppress the endotoxin production of microorganisms in the intestine, change the intestinal bacteria colony, and inhibit CD11b+ macrophages and CD11c+ dendritic cells to reduce body weight, body fat, blood sugar or inflammatory reactions. It is to provide a multifunctional new lactic acid bacteria that can control the back. In addition, the object of the present invention is to provide a variety of uses for new lactic acid bacteria.
  • the inventors of the present invention screen a myriad of lactic acid bacteria from kimchi to develop an anti-obesity, anti-diabetic or anti-inflammatory material with higher safety than synthetic chemicals, and among them, certain Lactobacillus strains inhibit endotoxin production of intestinal microorganisms And by changing the intestinal bacterial colonies and inhibiting CD11b+ cells, CD11c+ cells and CD11b+/CD11c+ cells in the intestine, it was discovered that it is possible to control body weight, body fat, blood sugar or inflammatory response, and completed the present invention.
  • an example of the present invention provides Lactobacillus sakei PK16 (Accession No.: KCCM12296P) as a novel lactic acid bacteria.
  • an example of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain, its culture, one or more selected from the crushed product or extract thereof It provides a composition for reducing body fat, including.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 (Accession No.: KCCM12296P) strain, its culture, a composition for lowering blood sugar comprising at least one selected from crushed products or extracts thereof as an active ingredient
  • an example of the present invention comprises at least one selected from Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, its culture, its crushed product, or an extract thereof, as an active ingredient, obesity, diabetes , Fatty liver, dyslipidemia, arteriosclerosis or at least two or more of the above diseases provides a composition used for the prevention, amelioration or treatment of metabolic syndrome.
  • an example of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain, its culture, crushing products, or endotoxemia comprising at least one selected from extracts thereof as an active ingredient prevention , To provide a composition for improvement or treatment.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 (Accession No.: KCCM12296P) strain, its culture, prevention of inflammatory diseases, including one or more selected from its crushing material or extract thereof as an active ingredient, Provides a composition for improvement or treatment.
  • an example of the present invention is a Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain, its culture, a composition for relieving stress comprising at least one selected from crushed products or extracts thereof as an active ingredient Gives
  • an example of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain, its culture, an anxiety disorder or cognition including at least one selected from its crushing material or extract thereof as an active ingredient
  • a composition for preventing, improving or treating dysfunction may be implemented in the form of pharmaceuticals or health functional food.
  • Certain Lactobacillus strains according to the present invention are separated from kimchi and have high safety, excellent anti-obesity activity, body fat reduction activity, blood sugar control activity, blood sugar lowering activity, blood cholesterol lowering activity, blood neutral lipid lowering activity, intestinal microflora It has various functions such as toxin production inhibitory activity, arteriosclerosis inhibitory activity, inflammation inhibitory activity, stress relieving activity, anti-anxiety activity, and cognitive function improving activity. In addition, certain Lactobacillus strains according to the present invention have excellent thermal stability.
  • Lactobacillus strains prevent, improve or treat obesity, diabetes, fatty liver, dyslipidemia, arteriosclerosis, metabolic syndrome, endotoxemia, inflammatory disease, stress, anxiety disorder, cognitive dysfunction, etc. It can be used as a material for pharmaceuticals or health functional foods.
  • FIG. 1 is a graph showing the effect of Lactobacillus sakei PK16 on the weight change of a model animal in which obesity is induced by a high fat diet for each experimental group
  • FIG. 2 is a body weight of a model animal in which obesity is induced by a high fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on the increase amount for each experimental group.
  • FIG. 3 is a graph showing the effect of Lactobacillus sakei PK16 on the weight change of epididymal adipose tissue of a model animal in which obesity is induced by a high fat diet by experimental group
  • FIG. 4 is a model animal in which obesity is induced by a high fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on the change in liver weight in each experiment group.
  • FIG. 5 is a graph showing the effect of Lactobacillus sakei PK16 on the plasma total cholesterol content of a model animal in which obesity is induced by a high-fat diet for each experimental group
  • FIG. 6 is a model animal in which obesity is induced by a high-fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on plasma neutral lipid of each group by experiment group.
  • FIG. 7 is a graph showing the effect of Lactobacillus sakei PK16 on the plasma endotoxin content of a model animal in which obesity is induced by a high-fat diet by each experimental group
  • FIG. 8 is a model animal in which obesity is induced by a high-fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on the fecal endotoxin content of each group.
  • FIG. 9 is a graph showing the effect of Lactobacillus sakei PK16 on the fasting glucose level of a model animal in which obesity is induced by a high-fat diet for each experimental group.
  • FIG. 10 is a graph showing the effect of Lactobacillus sakei PK16 on blood glucose changes in an oral glucose load test of a model animal in which obesity is induced by a high-fat diet by blood test group
  • FIG. 11 is an oral glucose load test of FIG. 10
  • the result is a graph represented by the area under the glucose-time curve (AUC).
  • FIG. 12 is a graph showing the effect of Lactobacillus sakei PK16 on colon length in each experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • FIG. 13 is a photograph showing the effect of Lactobacillus sakei PK16 on intestinal CD11b+ cells and intestinal CD11b+/CD11c+ cells in each experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • FIG. 14 is a graph showing the effect of Lactobacillus sakei PK16 on myeloperoxidase (Meloperoxidase, MPO) activity in colon tissues by experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • MPO myeloperoxidase
  • Figure 16 shows the anti-anxiety efficacy of Lactobacillus sakei PK16 in experimental animals stress-induced by a high-fat diet. The mouse stayed in the open arm among the results of the elevated plus-maze test. It is expressed as a percentage of time (OT, %).
  • FIG. 17 shows the anti-anxiety efficacy of Lactobacillus sakei PK16 in experimental animals in which stress was induced by a high-fat diet to the open arm of a mouse among the results of an elevated plus-maze test. It is expressed as the ratio of the number of accesses (OE, %).
  • FIG. 18 shows the efficacy of improving cognitive function of Lactobacillus sakei PK16 in experimental animals in which stress is induced by a high-fat diet as a result of a Y-maze test.
  • FIG. 19 shows the effect of improving the cognitive function of Lactobacillus sakei PK16 as a result of a novel object recognition test in experimental animals in which stress was induced by a high fat diet.
  • 20 is a graph showing the effect of Lactobacillus sakei PK16 on the corticosterone expression of stress-induced experimental animals by high-fat diet for each experimental group.
  • 21 is a graph showing the effect of Lactobacillus sakei PK16 on the expression of interleukin-6 (IL-6) in experimental animals stress-induced by a high-fat diet for each experimental group.
  • IL-6 interleukin-6
  • FIG. 22 shows the effect of Lactobacillus sakei PK16 on the expression of brain-derived neurotrophic factor (BDNF) and NF-kB activation (p-p65) in stress-induced experimental animals by high-fat diet. It is the result of measurement by blotting.
  • BDNF brain-derived neurotrophic factor
  • p-p65 NF-kB activation
  • culture means a product obtained by culturing a microorganism in a known liquid medium or solid medium, and is a concept in which the microorganism is included.
  • composition used in the present invention means a substance in which two or more components are uniformly mixed, and is a concept including an intermediate material for manufacturing a finished product as well as a finished product.
  • the terms “pharmaceutically acceptable” and “foodly acceptable” mean not significantly stimulating the organism and not inhibiting the biological activity and properties of the active substance administered.
  • prevention means all actions to suppress symptoms or delay progression of a specific disease by administration of the composition of the present invention.
  • treatment refers to any act that ameliorates or beneficially alters the symptoms of a particular disease by administration of a composition of the invention.
  • the term “improvement” refers to any action that at least reduces or alleviates the severity of the parameters associated with the condition being treated, such as symptoms.
  • the term “administration” means providing a subject composition of the present invention to an individual in any suitable way.
  • the individual refers to all animals, such as humans, monkeys, dogs, goats, pigs, or rats, who have a disease in which symptoms of a specific disease can be improved by administering the composition of the present invention.
  • the term "pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, which is the type, severity, activity of the drug, and the drug for the individual. Sensitivity, time of administration, route of administration and rate of discharge, duration of treatment, factors including the drugs used simultaneously, and other factors well known in the medical field can be determined.
  • One aspect of the present invention relates to a novel lactic acid bacteria having various functions such as anti-obesity activity, body fat reduction activity, blood sugar control activity, hypoglycemic activity, and the like.
  • the new lactic acid bacteria according to an example of the present invention is Lactobacillus sakei PK16 (Accession No.: KCCM12296P).
  • the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) is isolated from kimchi, has high safety, and has 16S rDNA consisting of the nucleotide sequence shown in SEQ ID NO:1.
  • the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) is a bacterium that is positive upon Gram staining, and L-arabinose, ribose, galactose, as a carbon source.
  • Glucose Fructose, Mannose, Rhamonse, N-acetyl-glucosamine, Esculin, Salicin, Cellobiose ), Lactose, Melibiose, Sucrose, Trehalose and Gluconate.
  • Lactobacillus sakei PK16 inhibits endotoxin production of intestinal microorganisms, changes intestinal bacterial colonies, and inhibits CD11b+ macrophages, CD11c+ dendritic cells and CD11b+/CD11c+ dendritic cells to reduce body weight, Body fat, blood sugar, or inflammatory reactions can be controlled.
  • the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) has anti-obesity activity, body fat reduction activity, blood sugar control activity, blood sugar lowering activity, blood cholesterol lowering activity, blood neutral lipid lowering activity, endotoxin production of intestinal microorganisms It has at least two or more activities selected from inhibitory activity, atherosclerotic inhibitory activity, anti-inflammatory activity, stress relief activity, anti-anxiety activity, or cognitive function improvement activity, and preferably at least three or more activities. Specifically, Lactobacillus sakei PK16 (Accession No.: KCCM12296P) increases Bacteroidetes in the intestinal bacterial community and decreases Firmicutes and Proteobacteria.
  • Firmicutes one of the gut microorganisms, is a bacterium that promotes sugar fermentation in the intestine and produces fatty acids to induce obesity.
  • a study published in the journal Nature in the British medical journal by Dr. Jeffrey Gordon of the University of Washington Genomics Science Center in the United States 'Fat people are bacteria in the intestines compared to slender people, and bacteria of the bacteroidetes family.
  • the human and animal gastrointestinal tracts contain more than 50 different phylums and more than 1000 different species of bacteria.
  • the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) has excellent thermal stability.
  • lactic acid bacteria when lactic acid bacteria are commercialized, they are powdered through lyophilization or spray drying, and the powdered lactic acid bacteria undergo various processes such as mixing with excipients and microencapsulation. Lactic acid bacteria may be exposed to a predetermined heat by various processing of the productization process, and further, may be exposed to a predetermined heat in the distribution or storage process of the product. Therefore, it can be said that the thermal stability of lactic acid bacteria is a very important factor in terms of commercialization of lactic acid bacteria. Therefore, Lactobacillus sakei PK16 of the present invention is evaluated as very advantageous from the viewpoint of productization and distribution/storage of the product.
  • an example of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain for use of new lactic acid bacteria, at least one selected from a culture thereof, a crushed product thereof, or an extract thereof. It provides a composition for reducing body fat, including.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 (Accession No.: KCCM12296P) strain, its culture, a composition for lowering blood sugar comprising at least one selected from crushed products or extracts thereof as an active ingredient
  • an example of the present invention comprises at least one selected from Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, its culture, its crushed product, or an extract thereof, as an active ingredient, obesity, diabetes , Fatty liver, dyslipidemia, arteriosclerosis or at least two or more of the above diseases provides a composition used for the prevention, amelioration or treatment of metabolic syndrome.
  • the metabolic syndrome refers to a disease in which multiple metabolic diseases such as diabetes and obesity appear simultaneously in one person, and narrowly refers to lipid-related metabolic syndrome.
  • the dyslipidemia refers to a condition in which total cholesterol, LDL cholesterol, and triglycerides are increased or HDL cholesterol is decreased in the blood, and one selected from the group consisting of hyperlipidemia, hypercholesterolemia and hypertriglyceridemia. It may be abnormal. Dyslipidemia is most often caused by obesity, diabetes, and alcohol.
  • the hyperlipidemia refers to a state in which lipids including cholesterol and triglycerides are increased in the blood. Hyperlipidemia can increase the risk of coronary artery disease, such as arteriosclerosis or myocardial infarction, even if it does not show symptoms.
  • the hypercholesterolemia is a condition in which cholesterol is increased in the blood, and total cholesterol and LDL cholesterol are high.
  • the hypertriglyceridemia refers to a state in which triglycerides are increased in
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 Accession No.: KCCM12296P
  • the endotoxemia refers to a condition in which endotoxin, a component of the cell wall of Gram-negative bacteria, is introduced into the blood, and there are infectious endotoxemia and non-infectious endotoxemia according to causes. Among them, non-infectious endotoxemia is known to originate from bacteria in the digestive tract.
  • the Lactobacillus sakei PK16 strain of the present invention is effective for preventing, improving or treating endotoxemia because it can inhibit the proliferation of intestinal microbes secreting endotoxins or inhibit the production of endotoxins in the intestinal microbes. It can be used as an ingredient.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 (Accession No.: KCCM12296P) strain, its culture, crushing products or extracts thereof.
  • a therapeutic composition If the inflammatory disease is a disease caused by an inflammatory reaction, the type is not particularly limited, and is preferably selected from atopic dermatitis, arthritis, or colitis.
  • the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain of the present invention reduces inflammation intestinal bacteria colonies and inhibits CD11b+ macrophages and CD11c+ dendritic cells that mediate inflammatory responses, thereby preventing inflammatory diseases , It can be used as an active ingredient for improvement or treatment.
  • An example of the present invention provides a composition for relieving stress comprising at least one selected from Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, its culture, crushed products thereof, or extracts thereof as an active ingredient do.
  • an example of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain, its culture, an anxiety disorder or cognition including at least one selected from its crushing material or extract thereof as an active ingredient
  • a composition for preventing, improving or treating dysfunction comprising at least one selected from Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, its culture, crushed products thereof, or extracts thereof as an active ingredient do.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 Accession No.: KCCM12296P
  • an anxiety disorder or cognition including at least one selected from its crushing material or extract thereof as an active ingredient
  • cognitive dysfunction means a symptom or a disease that does not normally exhibit cognitive functions such as memory processing, perception or problem solving, specifically working memory, attention and vigilance, language learning And memory, visual learning and memory, reasoning and problem solving, such as deterioration of executive function, processing speed or social cognition.
  • the cognitive dysfunction is mainly caused by impaired memory.
  • the culture is a product obtained by culturing a strain of genus Lactobacillus in a medium
  • the medium may be selected from known liquid medium or solid medium, for example, MRS liquid medium, MRS agar medium, BL agar medium.
  • the composition may be embodied as a pharmaceutical composition, a food additive, a food composition (especially a health functional food), or a feed additive, etc., depending on the purpose or aspect of use, a specific lactobacillus strain that is an active ingredient in the composition
  • the content of the light can also be adjusted in various ranges depending on the specific form of the composition, purpose of use, and aspect.
  • the content of the new lactic acid bacteria, its culture, its crushed material, or its extract, which is an active ingredient is not significantly limited, for example, 0.01 to 99% by weight based on the total weight of the composition, preferably 0.5 to It may be 50% by weight, more preferably 1 to 30% by weight.
  • the pharmaceutical composition according to the present invention may further include an additive such as a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient.
  • Carriers, excipients and diluents that may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • the pharmaceutical composition of the present invention is a known active ingredient having a prophylactic or therapeutic effect of obesity, diabetes, fatty liver, hyperlipidemia, cardiovascular disease, hypertension, arteriosclerosis or metabolic syndrome, in addition to novel lactic acid bacteria, cultures thereof, crushed products or extracts thereof It may contain one or more.
  • the pharmaceutical composition of the present invention is Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) strain as an active ingredient, one or more selected from Lactobacillus sake, a culture thereof, a crushed product thereof, or an extract thereof ( Lactobacillus sakei ) OK67 (Accession No.: KCCM11670P) may further include one or more selected from strains, cultures thereof, fragments thereof, or extracts thereof.
  • the pharmaceutical composition of the present invention may be formulated into a formulation for oral administration or a formulation for parenteral administration by conventional methods, and when formulated, a filler, a bulking agent, a binder, a wetting agent, a disintegrant, a surfactant, etc. It can be prepared using diluents or excipients.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, sucrose ), can be prepared by mixing Lactose or gelatin.
  • lubricants such as magnesium stearate talc may be used in addition to simple excipients.
  • Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions and syrups, but may include various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc. have.
  • Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories.
  • a non-aqueous solvent and a suspension solvent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.
  • composition of the present invention may be administered orally or parenterally to mammals, including humans, according to the desired method, and the parenteral administration methods include external application to the skin, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, and muscle. Intra-injection or intra-thoracic injection.
  • the dosage of the pharmaceutical composition of the present invention is not particularly limited as long as it is a pharmaceutically effective amount, and the range thereof depends on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity. Varies.
  • the usual daily dosage of the pharmaceutical composition of the present invention is not particularly limited, but is preferably 0.01 to 2000 mg/kg (body weight), more preferably 0.1 to 1000 mg/kg kg, based on the active ingredient. (body weight), and can be administered once or several times a day.
  • the content of new lactic acid bacteria, cultures thereof, crushed products or extracts thereof, which are active ingredients is 0.01 to 50% by weight, preferably 0.1 to 25% by weight, more preferably based on the total weight of the composition It is 0.5 to 10% by weight, but is not limited thereto.
  • the food composition of the present invention includes a form of pills, powders, granules, needles, tablets, capsules, or liquids, and examples of specific foods include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, Other dairy products including noodles, gums, and ice creams, various soups, beverages, teas, functional waters, drinks, alcoholic beverages, and vitamin complexes are included.Healthy functional foods and dietary supplements ( dietary supplements and food supplements.
  • the food composition of the present invention may contain, as an additional ingredient, a food-acceptable carrier, various flavoring agents, or natural carbohydrates, etc., in addition to the active ingredient.
  • the food composition of the present invention various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol , Carbonic acid used in carbonated beverages, and the like.
  • the food composition of the present invention may contain flesh for the preparation of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients can be used independently or in combination.
  • the above natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol.
  • the flavoring agent natural flavoring agents such as taumatin and stevia extract, synthetic flavoring agents such as saccharin and aspartame may be used.
  • an example of the present invention provides a method for reducing body fat by administering a Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain or composition to an individual in need.
  • an example of the present invention provides a method of controlling blood sugar or lowering blood sugar by administering a Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain or composition to an individual in need.
  • an example of the present invention by administering a strain or composition Lactobacillus sakei ( Lactobacillus sakei ) PK16 (Accession No.: KCCM12296P) to an individual in need, obesity, diabetes, fatty liver, dyslipidemia, arteriosclerosis or the above diseases It provides a method for preventing, improving or treating metabolic syndrome in which at least two or more of them occur simultaneously. In addition, an example of the present invention provides a method of preventing, improving or treating endotoxemia by administering a Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain or composition.
  • an example of the present invention provides a method of preventing, improving or treating inflammatory diseases by administering a Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain or composition.
  • the composition includes at least one selected from Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, culture thereof, crushed product or extract thereof, as an active ingredient, preferably pharmaceutically acceptable It may further include a carrier or a food acceptable carrier.
  • the composition may be administered orally or parenterally to mammals, including humans, depending on the desired method.
  • the typical daily dosage of the composition is not greatly limited, but preferably based on the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, preferably 1 ⁇ 10 9 to 1 ⁇ 10 12 CFU /Kg (body weight), more preferably 1 ⁇ 10 10 to 1 ⁇ 10 11 CFU/kg (body weight), and may be administered once or several times a day.
  • Lactobacillus sakei PK16 accesion No.: KCCM12296P
  • KCCM12296P Lactobacillus sakei PK16
  • the typical daily dosage of the composition is not greatly limited, but preferably based on the Lactobacillus sakei PK16 (Accession No.: KCCM12296P) strain, preferably 1 ⁇ 10 9 to 1 ⁇ 10 12 CFU /Kg (body weight), more preferably 1 ⁇ 10 10 to 1 ⁇ 10 11 CFU/kg (body weight), and may be administered once or several times a day.
  • Lactobacillus sakei OK67 Lactobacillus previously patented by the inventor of the present invention (accession number: KCCM11670P) and patent application, details are disclosed in Korean Registered Patent Publication No. 10-1708173
  • Lactobacillus sakei PK16 is an anaerobic bacterium showing positive when Gram staining, and carbon source availability among physiological properties is shown in Table 2 below.
  • Table 2 the carbon source availability of Lactobacillus sakei PK16 was analyzed by a sugar fermentation test by API Kit (model name: API 50 CHL; manufacturer: BioMerieux's, USA).
  • API Kit model name: API 50 CHL; manufacturer: BioMerieux's, USA.
  • “+” represents a case where the carbon source availability is positive
  • “-” represents a case where the carbon source availability is negative
  • " ⁇ " represents a case where the carbon source availability is ambiguous
  • the blank indicates unmeasured.
  • Lactobacillus four K (Lactobacillus sakei) PK16 as shown in Table 2, exhibited a very different carbon source utilization and Lactobacillus four K (Lactobacillus sakei) OK67.
  • 16S rDNA was measured by the chemical classification properties of Lactobacillus sakei PK16, and as a result, it was found to have the nucleotide sequence of SEQ ID NO: 1.
  • Lactobacillus sakei Lactobacillus sakei
  • 16K rDNA base sequence of PK16 was identified by BLAST search of Genebank (http://www.ncbi.nlm.nih.gov/).
  • Lactobacillus sake having the same 16S rDNA base sequence K ( Lactobacillus sakei ) strain was not detected, Lactobacillus sakei ( Lactobacillus sakei ) NBRC 15893 (NCBI ACCESSION: NR_113821) 16S rDNA partial sequence showed 99% homology and Lactobacillus sakei ( Lactobacillus sakei ) OK67 It showed 99% homology.
  • Lactic acid bacteria isolated from Chinese cabbage kimchi were pre-incubated in GAM (general anaerobic medium) liquid medium to prepare lactic acid bacteria samples.
  • E. coli was previously cultured in TSB (tryptic soy broth) medium to prepare an E. coli sample.
  • TSB tryptic soy broth
  • E. coli and any one selected from lactic acid bacteria isolated from Chinese cabbage kimchi are transplanted together in 10 ml of GAM (general anaerobic medium) liquid medium in an amount of 1 ⁇ 10 5 CFU and 1 ⁇ 10 5 CFU, respectively, at 37°C. Incubated for about 24 hr.
  • the culture medium was treated with ultrasound for about 1 hr to destroy the outer cell membrane of the bacteria, and centrifuged under conditions of 5000 ⁇ g to obtain a supernatant. Thereafter, the content of a representative endotoxin (lipopolysaccharide) present in the supernatant was measured by a LAL (Limulus Amoebocyte Lysate) assay kit (manufacturer: Cape Cod Inc., USA).
  • LAL Liimulus Amoebocyte Lysate
  • the cultured lactic acid bacteria were lyophilized and stored at 50° C. for 24 hr, transplanted into MRS agar medium, cultured in an anaerobic state to count the number of surviving bacteria. The survival rate was calculated.
  • Table 3 shows the results of measuring the inhibitory ability and heat stability of E. coli endotoxin production of lactic acid bacteria isolated from Chinese cabbage kimchi.
  • E. coli endotoxin production inhibitory ability -, less than 1%; +, 1% to 10%; ++, greater than 10% and up to 50%; +++, over 50%
  • Lactobacillus sakei OK67 was the best, followed by Lactobacillus sakei PK16.
  • Lactobacillus sakei PK6 and Lactobacillus sakei PK16 were the best.
  • Lactobacillus sakei OK67 showed very poor thermal stability.
  • lactic acid bacteria when lactic acid bacteria are commercialized, they are powdered through lyophilization or spray drying, and the powdered lactic acid bacteria undergo various processes such as mixing with excipients and microencapsulation. Lactic acid bacteria may be exposed to a predetermined heat by various processing of the productization process, and further, may be exposed to a predetermined heat in the distribution or storage process of the product. Therefore, it can be said that the thermal stability of lactic acid bacteria is a very important factor in terms of commercialization of lactic acid bacteria. Lactobacillus sakei PK16 of the present invention is evaluated to be very advantageous compared to Lactobacillus sakei OK67 from the viewpoint of commercialization because it is excellent in both E. coli endotoxin production inhibitory ability and thermal stability.
  • the low-fat diet group (NC group) was administered with a low-fat diet and physiological saline was administered orally for 4 weeks, and the experiment was terminated the next day.
  • the high-fat diet group was administered to the C group and oral administration of the physiological saline for 4 weeks, and the high-fat diet was administered to the L-PK16 group.
  • Lactobacillus sakei PK16 strain was orally administered for 4 weeks at a dose of 1 ⁇ 10 9 CFU/mouse once a day and the experiment was terminated the next day. Meanwhile, an oral glucose loading test was conducted 5 days before the end of the experiment.
  • the number of oral administration of the test sample containing the Lactobacillus sakei PK16 strain was 6 days based on 1 week.
  • the mice were kept in a breeding room environment with a temperature of 20 ⁇ 2°C, a humidity of 50 ⁇ 5% and a light-dark cycle of 12 hours.
  • epididymal fat (EF) weight, liver weight, plasma total cholesterol content, plasma neutral lipid content, plasma endotoxin, fecal endotoxin, and blood sugar were measured.
  • FIG. 1 is a graph showing the effect of Lactobacillus sakei PK16 on the weight change of a model animal in which obesity is induced by a high fat diet for each experimental group
  • FIG. 2 is a body weight of a model animal in which obesity is induced by a high fat diet
  • Figure 3 is a graph showing the effect of Lactobacillus sakei ( Lactobacillus sakei ) PK16 on the change in weight of epididymal adipose tissue of a model animal in which obesity was induced by a high fat diet for each experimental group
  • FIG. 4 is an obese induced by a high fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on the change in liver weight of model animals for each experimental group.
  • Plasma cholesterol, plasma trigLyceride, plasma endotoxin, and fecal endotoxin content of experimental animals induced obesity and administered lactic acid bacteria for 4 weeks were measured as follows.
  • Plasma total cholesterol was measured using a kit for measuring total cholesterol (ASAN PHARM. CO. LTD, Korea).
  • ASAN PHARM. CO. LTD, Korea a kit for measuring total cholesterol
  • the sample was heated for 10 minutes in a water bath at 37° C. with a standard solution having total cholesterol contents of 0, 75, 150, 225, and 300 mg/dl, respectively, and colorimetric was measured by measuring absorbance at 500 nm. It was quantified.
  • the plasma neutral lipid was measured using a neutral lipid measurement kit (ASAN PHARM. CO. LTD, Korea).
  • test tube 1.5 ml of the enzyme solution and 10 ⁇ l of plasma were added and stirred. Subsequently, the sample was heated for 10 minutes in a water bath at 37° C. with a standard solution having neutral lipid contents of 0, 75, 150, 225, and 300 mg/mm 2, respectively, and colorimetric determination by measuring absorbance at 550 nm. Did.
  • FIG. 5 is a graph showing the effect of Lactobacillus sakei PK16 on the plasma total cholesterol content of a model animal in which obesity is induced by a high-fat diet for each experimental group
  • FIG. 6 is a model animal in which obesity is induced by a high-fat diet
  • It is a graph showing the effect of Lactobacillus sakei PK16 on plasma neutral lipid of each group by experiment group.
  • the plasma total cholesterol was compared with the group in which only the high-fat diet after induction of obesity was fed. And plasma neutral lipid content.
  • Plasma endotoxin and fecal endotoxin content was measured using a Limulus amoebocyte lysate (LAL) Assay Kit (Cape Cod Inc., E. Falmouth, MA, USA).
  • LAL Limulus amoebocyte lysate
  • FIG. 8 is a model animal in which obesity is induced by a high-fat diet It is a graph showing the effect of Lactobacillus sakei PK16 on the fecal endotoxin content of each group.
  • LAL Limulus amoebocyte lysate
  • Endotoxin has a chemical structure that includes a polysaccharide and a lipid moiety, and is also chemically called a lipid polysaccharide (Lipopolysaccharide, hereinafter referred to as'LPS').
  • LPS' lipid polysaccharide
  • endotoxin Since one of the representative symptoms of inflammation is fever, endotoxin is also considered a pyrogen. However, if such an inflammatory reaction occurs excessively, the mechanism for body protection may be rather harmful, leading to endotoxemia, leading to death.
  • asthma is a typical disease associated with endotoxin in the living environment. Asthma is an allergic disease that occurs due to excessive immune responses, and continuous exposure to endotoxin through the respiratory tract causes chronic inflammation and develops as a final asthma. Moreover, it is known that asthma symptoms are exacerbated by exposure to endotoxin.
  • AUC 0.5 ⁇ (0.5 ⁇ C0+C15+C30+C60+C90+0.5 ⁇ C120)
  • C0, C15, C30, C60, C90, and C120 are glucose concentrations measured at 0, 15, 30, 60, 90, and 120 minutes respectively.
  • FIG. 9 is a graph showing the effect of Lactobacillus sakei PK16 on the fasting glucose level of a model animal in which obesity is induced by a high-fat diet for each experimental group.
  • FIG. 10 is a graph showing the effect of Lactobacillus sakei PK16 on blood glucose changes in an oral glucose load test of a model animal in which obesity is induced by a high-fat diet by blood test group
  • FIG. 11 is an oral glucose load test of FIG. 10
  • the result is a graph represented by the area under the glucose-time curve (AUC).
  • AUC glucose-time curve
  • the low-fat diet group (NC group) was administered with a low-fat diet and physiological saline was administered orally for 4 weeks, and the experiment was terminated the next day.
  • the high-fat diet group was administered to the C group and oral administration of the physiological saline for 4 weeks, and the high-fat diet was administered to the L-PK16 group.
  • Lactobacillus sakei PK16 strain was orally administered for 4 weeks at a dose of 1 ⁇ 10 9 CFU/mouse once a day and the experiment was terminated the next day.
  • the number of oral administration of the test sample containing the Lactobacillus sakei PK16 strain was 6 days based on 1 week.
  • the mice were kept in a breeding room environment with a temperature of 20 ⁇ 2°C, a humidity of 50 ⁇ 5% and a light-dark cycle of 12 hours.
  • the experimental animals were smothered with carbon dioxide, and the colon from the appendix to the site immediately before the anus was extracted and used.
  • CD11b+ macrophages and CD11c+ dendritic cells were measured by immunostaining.
  • Myeloperoxidase, MPO myeloperoxidase activity
  • 200 ⁇ l of 10 mM potassium phosphate buffer (pH 7.0) containing 0.5% hexadecyl trimethyl ammonium bromide was added to 100 mg of colon tissue and homogenized. Subsequently, the supernatant was obtained by centrifugation at 4° C. and 10,000 ⁇ g for 10 minutes. 50 ⁇ l of the supernatant was added to 0.95 ml of the reaction solution (containing 1.6 mM tetramethyl benzidine and 0.1 mM H 2 O 2 ), and the absorbance was measured over time at 650 nm while reacting at 37°C.
  • Myeloperoxidase (MPO) activity was calculated as 1 unit of 1 ⁇ mol/mL peroxide generated as a reactant.
  • Intestinal bacteria colonies were measured using qPCR (Quantitative PCR) method. Specifically, using PCR thermal cycler (Takara) and SYBR premix agent, qPCR (Quantitative PCR) was performed on the gut microbiota with 100 ng of DNA separated from the bowel movement (see Lee, Jeong, Han & Kim, 2018). ). The thermal cycle conditions for PCR are as follows.
  • Primer type Primer base sequence (5' ⁇ 3') Firmicutes Forward GGAGYATGTGGTTTAATTCGAAGCA Reverse AGCTGACGACAACCATGCAC Bacteroidetes Forward GTTTAATTCGATGATACGCGAG Reverse TTAASCCGACACCTCACGG ⁇ / ⁇ -Proteobacteria Forward GCTAACGCATTAAGTRYCCCG Reverse GCCATGCRGCACCTGTCT bacterial 16S rRNA Forward TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTGCCAGCMGCCGCGGTAA Reverse GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGACTACHVGGGTWTCTAAT
  • FIG. 12 is a graph showing the effect of Lactobacillus sakei PK16 on colon length in each experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • 13 is a photograph showing the effect of Lactobacillus sakei PK16 on intestinal CD11b+ cells and intestinal CD11b+/CD11c+ cells in each experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • 'DAPI' is a result of fluorescent staining with DAPI (4',6-diamidino-2-phenylindol)
  • 'Merge' is a combination of'CD11b'and'CD11c' results and DAPI (4',6- diamidino-2-phenylindol).
  • 14 is a graph showing the effect of Lactobacillus sakei PK16 on myeloperoxidase (Meloperoxidase, MPO) activity in colon tissues by experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • 15 is a result of analyzing the effect of Lactobacillus sakei PK16 on intestinal bacterial colonies in each experimental group in a model animal experiment in which colitis was induced by a high fat diet.
  • the Lactobacillus sakei PK16 strain suppressed the reduction of colon length by the high fat diet and decreased myeloperoxidase (MPO) activity increased by the high fat diet.
  • MPO myeloperoxidase
  • Inhibited CD11b+ cells (macrophages) and CD11b+/CD11c+ cells (dendritic cells) increased by high fat diet.
  • Lactobacillus sakei PK16 strain increases Bacteroidetes decreased by high fat diet among intestinal bacterial colonies and Firmicutes and Proteobacteria increased by high fat diet. Decreased. Therefore, Lactobacillus sakei ( Lactobacillus sakei ) PK16 strain has been shown to have a high anti-inflammatory effect and excellent in improving or treating colitis.
  • the low-fat diet group (NC group) was administered with a low-fat diet and physiological saline was administered orally for 4 weeks, and the experiment was terminated the next day.
  • the high-fat diet group was administered to the C group and the physiological saline was administered orally for 4 weeks, and the high-fat diet was administered to the L-PK16 group.
  • Lactobacillus sakei PK16 strain was orally administered for 4 weeks at a dose of 1 ⁇ 10 9 CFU/mouse once a day and the experiment was terminated the next day.
  • the number of oral administration of the test sample containing the Lactobacillus sakei PK16 strain was 6 days based on 1 week. During the experiment, the mice were kept in a breeding room environment with a temperature of 20 ⁇ 2°C, a humidity of 50 ⁇ 5% and a light-dark cycle of 12 hours.
  • interleukin-6 interleukin-6, IL-6
  • IL-6 interleukin-6
  • hippocampus BDNF (brain-derived neurotrophic factor)
  • hippocampus hippocampus NF-kB activation
  • p-p65 western blotting
  • the elevated plus maze device consists of two open arms (30 cm long, 7 cm wide) and two closed arms (30 cm long, 7 cm wide) facing the middle of the four aisles. Cm), each extending 7 cm wide and 7 cm long from the central platform.
  • the labyrinth was made of black Plexiglas and installed 50 cm above the floor.
  • a video camera was installed on the ceiling in the middle of the labyrinth to record the animal's behavior and the brightness was adjusted to 20 lux.
  • the mouse naturally likes to stay in the dark closed arm. So, when passing through the open arm, it shows considerable anxiety.
  • the mouse was placed with the head facing from the center to the open arm, and the maze was freely explored. Thereafter, the behavior of the mouse was observed for 5 minutes, and the arm entry was considered to have all 4 feet. After each experiment was over, wiped clean with 70% alcohol to remove any traces of all animals, so as not to affect the next experiment.
  • the ratio of the time the mouse stayed in the open arm (OT, %) and the ratio of the number of times the mouse went into and out of the open arm (OE, %) were calculated by the following equation.
  • the decrease in anxiety is indicated by an increase in the percentage of time spent in the open arm and an increase in the number of times it enters and exits the open arm.
  • the Y-maze device consists of three arms (8 cm wide, 30 cm long, 14 cm high) divided into A, B, and C, and each arm is 120° from each other. It is arranged at a constant angle. After placing the mouse on the end of one arm, allowing the Y-maze to move freely for 8 minutes, measure the number and sequence of the mouse entering each arm, and determine the spontaneous alteration (%) as follows. It was calculated by the formula.
  • the object object test Novel object recognition test
  • a box 40 cm wide, 40 cm long, 40 cm high
  • the mouse was started from the center of the box, and the number of times the mouse touched the two objects for 10 minutes was measured. Thereafter, after 24 hr, one of the two objects was replaced with a new object (B), and the number of times the original object and the new object were touched was measured.
  • the measured result was quantified as a preference index (%).
  • FIG. 16 shows the anti-anxiety efficacy of Lactobacillus sakei PK16 in experimental animals stress-induced by a high-fat diet. The mouse stayed in the open arm among the results of the elevated plus-maze test. It is expressed as a percentage of time (OT, %).
  • FIG. 17 shows the anti-anxiety efficacy of Lactobacillus sakei PK16 in experimental animals stress-induced by a high-fat diet, and the open arm of the mouse among the results of an elevated plus-maze test. It is expressed as a ratio (OE, %) of access to and from the city.
  • Figure 18 shows the efficacy of improving the cognitive function of Lactobacillus sakei ( Lactobacillus sakei ) PK16 in the stress-induced experimental animals by the high-fat diet as a result of the Y-maze test (Y-maze test).
  • Figure 19 shows the effect of improving the cognitive function of Lactobacillus sakei ( Lactobacillus sakei ) PK16 in the experimental animal stress-induced by high-fat diet as a result of the object recognition test (Novel object recognition test).
  • 20 is a graph showing the effect of Lactobacillus sakei PK16 on the corticosterone expression of stress-induced experimental animals by high-fat diet for each experimental group.
  • FIG. 21 is a graph showing the effect of Lactobacillus sakei PK16 on the expression of interleukin-6 (IL-6) in experimental animals stress-induced by a high-fat diet for each experimental group.
  • FIG. 22 shows the effect of Lactobacillus sakei PK16 on the expression of brain-derived neurotrophic factor (BDNF) and NF-kB activation (p-p65) in experimental animals stress-induced by high-fat diet. It is the result of measurement by blotting.
  • BDNF brain-derived neurotrophic factor
  • p-p65 NF-kB activation
  • Lactobacillus sakei PK16 strain significantly suppressed anxiety caused by a high fat diet.
  • Lactobacillus sakei PK16 strain significantly improved cognitive impairment caused by a high fat diet.
  • the Lactobacillus sakei PK16 strain increased the content of corticosterone and interleukin-6 (interleukin-6, IL-6) in the blood increased by a high fat diet. Suppressed.
  • Lactobacillus sakei Lactobacillus sakei
  • PK16 strain suppressed increased hippocampus (hippocampus) NF-kB activation (p-p65) by high fat diet, whereas hippocampus inhibited by high fat diet (hippocampus) ) BDNF (brain-derived neurotrophic factor) expression was increased.
  • Lactobacillus sakei PK16 culture can be replaced with Lactobacillus sakei PK16 strain itself, its lysate or extract thereof.
  • Lactobacillus sakei PK16 culture was added to 100 parts by weight of wheat flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.
  • Lactobacillus sakei PK16 culture 0.5 parts by weight was added to 100 parts by weight of milk, and various dairy products such as butter and ice cream were prepared using the milk.
  • the brown rice, barley, glutinous rice, and yulmu were alpha-polished by a known method to distribute the dried one, and then prepared into a powder having a particle size of 60 mesh with a grinder.
  • Black soybeans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, and then distributed into a powder having a particle size of 60 mesh with a grinder.
  • the above-prepared grains, seeds, and Lactobacillus sakei PK16 cultures were prepared by mixing in the following proportions.
  • Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds),
  • Liquid fructose 0.5 g
  • oligosaccharide 4 g
  • sugar 2 g
  • table salt 0.5 g
  • water 77 g
  • other ingredients such as Lactobacillus sakei PK16 culture were homogeneously blended for instant sterilization. Then, it was manufactured by packaging it in small packaging containers such as glass bottles and plastic bottles.
  • Vegetable juice was prepared by adding 2 g of Lactobacillus sakei PK16 culture to 1,000 ml of tomato or carrot juice.
  • Fruit juice was prepared by adding 1 g of Lactobacillus sakei PK16 culture to 1,000 ml of apple or grape juice.
  • the inventors of the present invention patented Lactobacillus sakei PK16 on August 03, 2018 at the Korea Microbiological Conservation Center (3F, Eurim Building, 45, Hongjenae 2-ga-gil, Seodaemun-gu, Seoul, Korea) and deposited KCCM12296P. You have been given a number.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Diabetes (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • General Engineering & Computer Science (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biochemistry (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Urology & Nephrology (AREA)
  • Vascular Medicine (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Nutrition Science (AREA)

Abstract

La présente invention concerne une nouvelle souche de Lactobacillus isolée à partir du kimchi. La souche de Lactobacillus particulière, selon la présente invention, est isolée du kimchi et présente ainsi une sécurité élevée et diverses fonctions telles qu'une excellente activité anti-obésité, l'activité de réduction de la graisse corporelle, l'activité de régulation du glucose sanguin, l'activité de réduction du glucose sanguin, l'activité de réduction du cholestérol sanguin, l'activité de réduction des lipides neutres sanguins, l'activité inhibitrice de la production d'endotoxines du microbiote intestinal, l'activité inhibitrice de l'artériosclérose, l'activité anti-inflammatoire, l'activité de soulagement du stress, l'activité anti-anxiété, l'activité d'amélioration de la fonction cognitive, etc. La souche de Lactobacillus particulière selon la présente invention présente également une excellente stabilité thermique. Par conséquent, la souche de Lactobacillus particulière selon la présente invention peut être utilisée en tant que constituant pour un médicament ou un aliment naturel fonctionnel pour prévenir, apporter des améliorations à ou traiter l'obésité, le diabète, la stéatose hépatique, la dyslipidémie, l'artériosclérose, le syndrome métabolique, l'endotoxémie, une maladie inflammatoire, le stress, un trouble anxieux, le dysfonctionnement cognitif, etc.
PCT/KR2019/017441 2018-12-18 2019-12-11 Nouveau lactobacillus ayant un effet de réduction du poids corporel ou de la graisse corporelle et son utilisation WO2020130471A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020180164469A KR20200076001A (ko) 2018-12-18 2018-12-18 체중 및 체지방 감소 효능을 갖는 신규 유산균 및 이의 용도
KR10-2018-0164469 2018-12-18

Publications (1)

Publication Number Publication Date
WO2020130471A1 true WO2020130471A1 (fr) 2020-06-25

Family

ID=71101852

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2019/017441 WO2020130471A1 (fr) 2018-12-18 2019-12-11 Nouveau lactobacillus ayant un effet de réduction du poids corporel ou de la graisse corporelle et son utilisation

Country Status (2)

Country Link
KR (1) KR20200076001A (fr)
WO (1) WO2020130471A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114292795A (zh) * 2022-01-20 2022-04-08 中国疾病预防控制中心传染病预防控制所 乳酸杆菌益生菌CGMCC No.1.13855及在制备降脂药物中的应用
CN114317326A (zh) * 2021-12-06 2022-04-12 浙江工商大学 一种用于缓解过敏反应的清酒乳杆菌及制备方法
US20220118034A1 (en) * 2019-02-22 2022-04-21 Korea Food Research Institute Kimchi lactobacillus sakei having prophylactic, ameliorating or therapeutic effect on depression and anxiety disorders
CN114561320A (zh) * 2022-01-20 2022-05-31 中国疾病预防控制中心传染病预防控制所 乳酸杆菌益生菌CGMCC No.1.13855在制备肝病治疗药物中的应用
CN115029270A (zh) * 2022-06-21 2022-09-09 江南大学 一株能够降低肠道促炎细胞因子的清酒乳杆菌及其应用
CN115181695A (zh) * 2022-06-27 2022-10-14 广东粤港澳大湾区国家纳米科技创新研究院 一种植物乳杆菌5b4m2及其应用
CN118272276A (zh) * 2024-05-24 2024-07-02 吉林农业大学 一种清酒广布乳杆菌及其产品在降低血糖中的应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20160097183A (ko) * 2014-02-17 2016-08-17 경희대학교 산학협력단 비만 억제 효능을 갖는 신규 락토바실러스 사케이 및 이의 용도
KR101708173B1 (ko) * 2015-03-06 2017-02-21 경희대학교 산학협력단 혈당 조절 효능을 갖는 신규 유산균 및 이의 용도
KR20170032815A (ko) * 2015-09-15 2017-03-23 경희대학교 산학협력단 신규 유산균 및 퇴행성 뇌질환 또는 인지기능의 예방, 개선 또는 치료용 조성물
KR101736033B1 (ko) * 2016-05-09 2017-05-17 한국식품연구원 항비만 활성을 가지는 락토바실러스 사케아이
KR20180036592A (ko) * 2016-09-30 2018-04-09 씨제이제일제당 (주) 신규한 락토바실러스 사케이 및 이를 포함하는 조성물

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20160097183A (ko) * 2014-02-17 2016-08-17 경희대학교 산학협력단 비만 억제 효능을 갖는 신규 락토바실러스 사케이 및 이의 용도
KR101708173B1 (ko) * 2015-03-06 2017-02-21 경희대학교 산학협력단 혈당 조절 효능을 갖는 신규 유산균 및 이의 용도
KR20170032815A (ko) * 2015-09-15 2017-03-23 경희대학교 산학협력단 신규 유산균 및 퇴행성 뇌질환 또는 인지기능의 예방, 개선 또는 치료용 조성물
KR101736033B1 (ko) * 2016-05-09 2017-05-17 한국식품연구원 항비만 활성을 가지는 락토바실러스 사케아이
KR20180036592A (ko) * 2016-09-30 2018-04-09 씨제이제일제당 (주) 신규한 락토바실러스 사케이 및 이를 포함하는 조성물

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
JANG, H.-M.: "Lactobacillus sakei alleviates high-fat-diet-induced obesity and anxiety in mice by inducing AMPK activation and SIRT1 expressior and inhibiting gut microbiota-mediated NF-kappaB activation", MOLECULAR NUTRITION & FOOD RESEARCH, vol. 63, 11 February 2019 (2019-02-11), pages 1800978 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20220118034A1 (en) * 2019-02-22 2022-04-21 Korea Food Research Institute Kimchi lactobacillus sakei having prophylactic, ameliorating or therapeutic effect on depression and anxiety disorders
CN114317326A (zh) * 2021-12-06 2022-04-12 浙江工商大学 一种用于缓解过敏反应的清酒乳杆菌及制备方法
CN114317326B (zh) * 2021-12-06 2023-09-12 浙江工商大学 一种用于缓解过敏反应的清酒乳杆菌及制备方法
CN114292795A (zh) * 2022-01-20 2022-04-08 中国疾病预防控制中心传染病预防控制所 乳酸杆菌益生菌CGMCC No.1.13855及在制备降脂药物中的应用
CN114561320A (zh) * 2022-01-20 2022-05-31 中国疾病预防控制中心传染病预防控制所 乳酸杆菌益生菌CGMCC No.1.13855在制备肝病治疗药物中的应用
CN114292795B (zh) * 2022-01-20 2023-07-25 中国疾病预防控制中心传染病预防控制所 乳酸杆菌益生菌CGMCC No.1.13855及在制备降脂药物中的应用
CN115029270A (zh) * 2022-06-21 2022-09-09 江南大学 一株能够降低肠道促炎细胞因子的清酒乳杆菌及其应用
CN115029270B (zh) * 2022-06-21 2023-11-28 江南大学 一株能够降低肠道促炎细胞因子的清酒乳杆菌及其应用
CN115181695A (zh) * 2022-06-27 2022-10-14 广东粤港澳大湾区国家纳米科技创新研究院 一种植物乳杆菌5b4m2及其应用
CN115181695B (zh) * 2022-06-27 2023-08-29 广东粤港澳大湾区国家纳米科技创新研究院 一种植物乳杆菌5b4m2及其应用
CN118272276A (zh) * 2024-05-24 2024-07-02 吉林农业大学 一种清酒广布乳杆菌及其产品在降低血糖中的应用
CN118272276B (zh) * 2024-05-24 2024-08-23 吉林农业大学 一种清酒广布乳杆菌及其产品在降低血糖中的应用

Also Published As

Publication number Publication date
KR20200076001A (ko) 2020-06-29

Similar Documents

Publication Publication Date Title
WO2020130471A1 (fr) Nouveau lactobacillus ayant un effet de réduction du poids corporel ou de la graisse corporelle et son utilisation
WO2017204374A1 (fr) Nouvelles bactéries lactiques capables de réguler la glycémie et leur utilisation
WO2016053003A1 (fr) Procédé pour la préparation d'une préparation microbienne dans lequel de l'aglycone s'accumule dans des cellules et préparation microbienne préparée par ce procédé
WO2015122717A1 (fr) Nouvelles bactéries d'acide lactique possédant un effet inhibiteur sur l'obésité et leur utilisation
WO2017131402A1 (fr) Nouvelle bactérie lactique dérivée d'intestin humain ayant une fonction immunorégulatrice, et son utilisation
KR101708173B1 (ko) 혈당 조절 효능을 갖는 신규 유산균 및 이의 용도
WO2021194225A1 (fr) Souche de lactobacillus delbrueckii subsp. lactis ckdb001 et composition de prévention d'amélioration ou de traitement de stéatose hépatique non alcoolique la comprenant
WO2011010770A1 (fr) Nouvelle souche de lactobacillus plantarum et composition la contenant
WO2019088379A1 (fr) Nouvelles bactéries lactiques et leur utilisation
WO2011007922A1 (fr) Nouveau lactobacillus plantarum et composition en contenant
WO2018062914A1 (fr) Nouveau lactobacillus sakei et composition le comprenant
WO2023055188A1 (fr) Nouveaux probiotiques et utilisation associée
WO2017047962A1 (fr) Nouveau lactobacillus et composition pour prévenir, atténuer ou traiter des maladies cérébrales dégénératives ou des troubles de la fonction cognitive
WO2022145807A1 (fr) Nouvelle souche de bifidobacterium animalis subsp. lactis hem20-01 et composition pour le traitement de la dépression la comprenant ou comprenant une culture de celle-ci
WO2020262755A1 (fr) Nouvelle composition probiotique pour la régulation de l'immunité intestinale
WO2019098810A2 (fr) Nouvelles bactéries lactiques et leur utilisation
WO2020116962A1 (fr) Composition de prévention, d'amélioration, ou de traitement du diabète, comprenant le produit de fermentation de bactéries acide lactique d'extrait de paeonia lactiflora comme ingrédient actif
KR102570432B1 (ko) 락토바실러스 플란타룸 k97 및 이의 용도
WO2010064778A1 (fr) Nouveau lactobacillus plantarum et composition le contenant
WO2022005035A1 (fr) Nouvelle souche de bifidobacterium breve idcc 4401 et sa cellule morte id-bbr4401 ayant d'excellentes tolérances aux acides et à la bile et un effet prophylactique ou thérapeutique sur la dyslipidémie
WO2024136036A1 (fr) Composition pour la prévention ou le traitement de l'arthrite, contenant lactobacillus brevis ku15147
US20230107022A1 (en) Lactobacillus rhamnosus lrh05 isolate, and composition including the same and use thereof
WO2023229394A1 (fr) Souche de lactobacillus paracasei ou souche de lactobacillus plantarum issue du corps humain, présentant une activité de réduction de la graisse corporelle, et composition de mélange la comprenant
KR20180075463A (ko) 프로바이오틱 활성을 갖는 신규 락토바실러스 퍼멘텀 및 이의 용도
WO2022265431A1 (fr) Utilisation d'une souche de lactobacillus fermentum et de lymphocytes t régulateurs en polythérapie pour la prévention et le traitement d'une maladie métabolique

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19897987

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19897987

Country of ref document: EP

Kind code of ref document: A1