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WO2018111023A1 - Bactéries lactiques présentant une stabilité accrue, et leur procédé de production - Google Patents

Bactéries lactiques présentant une stabilité accrue, et leur procédé de production Download PDF

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Publication number
WO2018111023A1
WO2018111023A1 PCT/KR2017/014806 KR2017014806W WO2018111023A1 WO 2018111023 A1 WO2018111023 A1 WO 2018111023A1 KR 2017014806 W KR2017014806 W KR 2017014806W WO 2018111023 A1 WO2018111023 A1 WO 2018111023A1
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Prior art keywords
lactic acid
acid bacteria
cfu
coating
weight
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Korean (ko)
Inventor
엄기안
연성흠
손락호
박채리
필감방
윤지수
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Huons Co Ltd
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Huons Co Ltd
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Priority claimed from KR1020170172396A external-priority patent/KR102004204B1/ko
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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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • 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
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P20/00—Coating of foodstuffs; Coatings therefor; Making laminated, multi-layered, stuffed or hollow foodstuffs
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • 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
    • 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

Definitions

  • the present invention relates to a lactic acid bacterium having improved stability and a method for preparing the same, more specifically xylitol; Gelatin or collagen; And the present invention relates to a lactic acid bacterium and a method for producing the same which is enhanced by storage and thermal stability, acid resistance, bile resistance, and digestive enzymes by triple coating of dextrin sequentially.
  • Lactic acid bacteria also known as lactic acid bacteria or lactic acid bacteria, are gram-positive bacteria that break down sugars such as glucose to produce lactic acid. They are found in the digestive tract, oral cavity, and vagina of humans and mammals. Lactic acid bacteria are one of the most widely used microorganisms for a long time, and do not produce harmful substances to the intestines of humans or animals, and are useful as a formal preparation by preventing abnormal fermentation by harmful bacteria in the intestines. In addition, the lactic acid produced by the lactic acid fermentation has been used to produce foods such as dairy products, kimchi, brewed foods, etc., which inhibit the growth of pathogens and harmful bacteria.
  • probiotics are living bacteria that enter the body and give a healthy effect.
  • Most probiotics known to date are Lactobacillus sp . , Lactococcus sp . ), Streptococcus sp . , Leuconostoc sp . ), And O Phedi deulyimyeo lactic acid bacteria belonging to genus Lactococcus such as (Pedicoccus sp.), Includes a part of Bacillus (Bacillus) and the like.
  • the function of lactic acid bacteria and probiotics has been studied for a long time since it was found by Ilya Mechinnikov that Bulgarians enjoy longevity due to the consumption of fermented milk fermented with Lactobacillus.
  • probiotics are reported to be effective in preventing lactose intolerance and colon cancer, lowering cholesterol and blood pressure, improving immune function, preventing infection, improving mineral absorption, growing harmful bacteria due to stress, and improving irritable bowel syndrome and colitis. It has also been recommended as a treatment and antibiotic for strengthening the immune system and for treating intestinal diseases associated with candidiasis.
  • microorganisms including lactic acid bacteria must survive in gastric acid and bile acids, reach the small intestine, proliferate and settle in the intestine, exhibit useful effects in the intestine, be nontoxic and non-pathogenic.
  • Lactic acid bacteria food is made of powder, granules, tablets, capsules, etc. to cultivate live bacteria such as lactobacillus, lactobacillus, bifidus and other foods in a stable and easy way.
  • Lactic acid bacteria production process as described above is largely divided into lactic acid bacteria culture, cell recovery, lyophilization, grinding, commercialization, etc.
  • lactic acid bacteria are exposed to various physicochemical stresses.
  • the cell recovery is affected by osmotic pressure due to concentration, and during freeze-drying process, ice crystals in the cytoplasm are formed due to rapid temperature change, or ice crystals and dehydration generated outside the cell affect the temperature and osmotic pressure.
  • osmotic pressure due to concentration
  • ice crystals in the cytoplasm are formed due to rapid temperature change, or ice crystals and dehydration generated outside the cell affect the temperature and osmotic pressure.
  • lactic acid bacteria is reduced when exposed to high temperature, high pressure or hydrated by moisture in the air during grinding and commercialization, and liquid products such as lactic acid bacteria fermented foods, lactic acid bacteria fermented milk, lactic acid bacteria beverages stored for a short period of time, as well as long-term Even in products manufactured in powder form for the purpose of storage, it has been pointed out that when exposed to oxygen, the fatty acids constituting the cell membrane are oxidized to reduce the survival rate.
  • probiotics products utilize the live bacteria themselves, they are exposed to various stresses in the human body before reaching the intestines after ingestion.
  • the survival rate may be greatly reduced by being exposed to an acidic environment falling below pH 3 and affected by digestive enzymes and bile acids secreted in the small intestine. Even when it reaches the intestine, it competes with the microorganisms that have settled in the intestine, and at the same time, the growth is inhibited by various harmful components and active oxygen.
  • a high concentration of lactic acid bacteria is introduced a method of producing a double-structured jelly or double coating with protein and polysaccharides, or direct reaction with air, water is suppressed and heat resistance, acid resistance and A method of preparing tri-coated lactic acid bacteria (BK Patent Application No. 10-2008-10397) with enhanced bile properties and enhanced viability and processing stability has been developed.
  • the coating technique of the conventional lactic acid bacteria also can not completely coat the surface of the lactic acid bacteria, it is still pointed out that the lactic acid bacteria prepared by the above method is not sufficiently excellent in heat resistance, acid resistance and bile resistance.
  • the present inventors earnestly researched to develop a method capable of improving the storage and lactic acid bacteria stability in the body, as a result, the lactic acid bacteria, preferably Lactobacillus pentosus strain Xylitol; Gelatin or collagen; And the result of the three-coating sequentially using the dextrin was confirmed that the storage stability, thermal stability, acid resistance, bile resistance, and the stability of the digestive enzymes of the strain is improved, the present invention was completed based on this.
  • the lactic acid bacteria preferably Lactobacillus pentosus strain Xylitol
  • Gelatin or collagen the result of the three-coating sequentially using the dextrin was confirmed that the storage stability, thermal stability, acid resistance, bile resistance, and the stability of the digestive enzymes of the strain is improved, the present invention was completed based on this.
  • an object of the present invention is to provide a method for producing lactic acid bacteria with improved stability.
  • the present invention provides a method for producing lactic acid bacteria with improved stability, including the following steps.
  • the present invention provides a method for enhancing the stability of lactic acid bacteria, comprising the following steps.
  • xylitol in the step (a) may be 1% to 30% concentration.
  • the gelatin or collagen in step (b) may be 1% to 30% concentration.
  • the dextrin in step (c) may be a concentration of 10% to 50%.
  • the xylitol in the step (a), is mixed with the primary coating in a ratio of 10 parts by weight to 100 parts by weight with respect to 100 parts by weight of lactic acid bacteria cells;
  • the gelatin or collagen is mixed with a secondary coating in a ratio of 0.5 parts by weight to 50 parts by weight with respect to 100 parts by weight of lactic acid bacteria cells;
  • the dextrin may comprise a third coating by mixing in a ratio of 0.5 parts by weight to 50 parts by weight with respect to 100 parts by weight of lactic acid bacteria cells.
  • the lactic acid bacteria is Lactobacillus sp. , Genus Bifidobacterium ( Bifidobacterium) sp . ), Genus Streptococcus sp. , Genus Lactococcus sp . ) Enterococcus sp . ), Pediococcus sp . , Genus Leuconostoc sp . ), And Weissella sp. ) May be one or more strains selected from the group consisting of.
  • the lactic acid bacteria may be Lactobacillus pentosus .
  • Lactic acid bacteria prepared by the coating method according to the present invention is xylitol; Gelatin or collagen; And triple-coating sequentially with dextrin, to experimentally confirm that the storage temperature and heat stability, acid resistance, bile resistance, and digestive enzyme stability are improved compared to lactic acid bacteria which have not been conventionally coated, thereby resisting external stress. And it was confirmed that the viability of the strain when passing through the gastrointestinal tract and can maintain the original physiological activity function in the intestine, the coating method according to the present invention is fermented milk, fermented food, functional food, general food, cosmetics, and using lactic acid bacteria It is expected to be usefully used in related industries such as pharmaceuticals.
  • 1 is a photograph of the final raw material after freeze-drying the Lactobacillus pentosus KF340 strain sequentially coated with xylitol, gelatin, and dextrin triple.
  • Figure 2 is a photograph of the final raw material after lyophilization of Lactobacillus pentosus KF340 strain sequentially coated with xylitol, collagen, and dextrin in threefold.
  • Figure 3 is a photograph taken with a scanning electron microscope of the Lactobacillus pentosus KF340 strain sequentially coated with xylitol, gelatin, and dextrin triple.
  • FIG. 4 is a photograph taken with a scanning electron microscope of the Lactobacillus pentosus KF340 strain sequentially coated with xylitol, collagen, and dextrin in a triple.
  • the present inventors have studied diligently to develop a coating method that can enhance the storage and stability of the lactic acid bacteria in the body, and as a result, the storage and thermal stability of the strain, acid resistance, bile resistance, and lactic acid bacteria with enhanced stability to digestive enzymes
  • the present invention has been completed by developing a coating method.
  • the present invention (a) adding xylitol to the lactic acid bacteria and homogenizing the first coating; (b) adding a gelatin or collagen to the first coated lactic acid bacteria and homogenizing the second coating to homogenize; And (c) adding a dextrin (dextrin) to the secondary coated lactic acid bacteria and provides a method of producing a lactic acid bacteria with improved stability, comprising the step of homogeneous tertiary coating.
  • the present invention (a) adding xylitol to the lactic acid bacteria and homogenizing the first coating; (b) adding a gelatin or collagen to the first coated lactic acid bacteria and homogenizing the second coating to homogenize; And (c) adding a dextrin to the secondary coated lactic acid bacteria and homogenizing the third coating to provide a method for enhancing stability of the lactic acid bacteria.
  • the lactic acid bacteria culture can be carried out in conventional lactic acid culture medium and culture conditions according to the type of lactic acid bacteria to be used, and is not particularly limited.
  • xylitol is dissolved in distilled water and may be in a concentration of 1% to 30%, preferably 1% to 15%, more preferably 3% to 7%, even more preferably 5%, and use. Pre-sterilized, cooled to room temperature and available for coating.
  • the xylitol is added to 10 parts by weight to 100 parts by weight, preferably 50 parts by weight to 100 parts by weight, more preferably 100 parts by weight, which is the same amount as that of recovered cells, and homogenized, and room temperature.
  • the cells can be first coated by standing at room temperature for 0.5 to 3 hours, more preferably 1 hour to 2 hours.
  • gelatin may be dissolved in distilled water at a concentration of 1% to 30%, preferably 1% to 15%, more preferably 4% to 7%, even more preferably 6%, and used. Pre-sterilized, cooled to room temperature and available for coating.
  • the gelatin is added to 0.5 parts by weight to 50 parts by weight, preferably 5 parts by weight to 50 parts by weight, more preferably 10 parts by weight to 30 parts by weight, based on 100 parts by weight of the cells recovered after the culture, and homogenized at room temperature.
  • the cells can be secondary coated by standing at room temperature for 0.5 to 3 hours, more preferably 1 to 2 hours.
  • the collagen may be dissolved in distilled water at a concentration of 1% to 30%, preferably 5% to 20%, more preferably 10% to 15%, even more preferably 12%, and used. Pre-sterilized, cooled to room temperature and available for coating.
  • the collagen is homogenized after adding 0.5 parts by weight to 50 parts by weight, preferably 5 parts by weight to 50 parts by weight, more preferably 10 parts by weight to 30 parts by weight, based on 100 parts by weight of the cells recovered after the culture.
  • the cells can be secondary coated by standing at room temperature for 0.5 to 3 hours, more preferably 1 to 2 hours.
  • the dextrin may be dissolved in distilled water at a concentration of 10% to 50%, preferably 10% to 30%, more preferably 15% to 25%, even more preferably 20%, and used. Pre-sterilized, cooled to room temperature and available for coating.
  • the dextrin is homogenized after adding 0.5 parts by weight to 50 parts by weight, preferably 5 parts by weight to 50 parts by weight, more preferably 10 parts by weight to 30 parts by weight, based on 100 parts by weight of the cells recovered after the culture.
  • the cells may be tertiary coated by standing at room temperature for 0.5 to 3 hours, more preferably 1 to 2 hours.
  • the present invention may include a process of lyophilizing the coated lactic acid bacteria after the lactic acid bacteria coating process as described above, it can be carried out by those skilled in the art according to a conventional method.
  • the stability is meant to include both in vitro stability and in vivo stability to lactic acid bacteria, more specifically, storage and heat resistance to lactic acid bacteria, bile acid, acid resistance, and digestive enzymes Stability, and stability in intestinal conditions.
  • the stability of the strain was evaluated in various aspects as compared to the case where the Lactobacillus pentosus strain was not coated after the triple coating by the above production method.
  • the viable cell count and viability were measured at two-week intervals while storing the lactic acid bacteria samples for 4 weeks at 4, 25, and 40 ° C. conditions to evaluate storage stability enhancement.
  • the rate of decrease of the cells increases, and it was confirmed that the survival rate of the tri-coated lactic acid bacteria was significantly increased as compared with the case without the coating treatment (see Example 3).
  • the lactic acid bacteria samples were treated with artificial gastric juice of pH 2.0 and 2.5, respectively, and reacted for 30 minutes, 60 minutes, and 120 minutes, and then the viable cell count and survival rate. Measured. As a result, it was found that the survival rate was higher in the triple coated sample according to the present invention compared to the case where the coating was not performed (see Example 4).
  • the lactic acid bacteria were treated with 0.2% pancreatin and 100 unit / mL of alpha-amylase, respectively, for 30 minutes, 60 minutes, and to evaluate the stability of the lactic acid bacteria for digestive enzymes. After reacting for 120 minutes, viable cell count and viability were measured. As a result, it was possible to confirm the high digestive enzyme stability of the Lactobacillus pentosus strain itself used in this example, it was confirmed that the stability of the digestive enzymes of the lactic acid bacteria by the triple coating treatment according to the present invention is more enhanced. (See Example 6).
  • the triple coating method according to the present invention generally improves the storage stability, acid resistance, bile resistance, stability against digestive enzymes, and thermal stability according to storage temperature, compared to the non-coated lactic acid bacteria.
  • the triple coating method according to the present invention generally improves the storage stability, acid resistance, bile resistance, stability against digestive enzymes, and thermal stability according to storage temperature, compared to the non-coated lactic acid bacteria.
  • the present invention provides a tri-coated lactic acid bacteria prepared by the above method.
  • the lactic acid bacteria are Lactobacillus bacteria (Lactobacillus sp.) Genus, Bifidobacterium (Bifidobacterium sp.), A Streptococcus (Streptococcus sp.) Genus Lactococcus (Lactococcus sp.) Genus Enterococcus (Enterococcus sp . ), Pediococcus sp . , Genus Leuconostoc sp . ), And Weissella sp.
  • Genus may be one or more strains selected from the group consisting of, more preferably Lactobacillus pentosus ( Lactobacillus pentosus ), more preferably Lactobacillus pentosus KF340 (Accession number KCCM 11675P).
  • Lactobacillus pentosus KF340 strain was incubated with stirring for 12 ⁇ 3 hours at 32 °C, pH 6.0 conditions using 360 L modified-MRS medium.
  • the cultured Lactobacillus pentosus KF340 cells were separated using a centrifuge, and 5.2 Kg of cells were recovered through this process.
  • Experimental group 1 is a triple coating strain using xylitol, gelatin, and dextrin
  • experimental group 2 is a triple coating strain using xylitol, collagen, and dextrin
  • the control group is an uncoated strain.
  • Experimental Group 1 dissolved each raw material in distilled water so as to have a concentration of 5% xylitol, 6% gelatin, and 20% dextrin. Sterilized at 121 ° C. for 15 minutes and then cooled at room temperature. Next, the same amount of 5% xylitol aqueous solution was added to the recovered cells, and the cells were homogenized and allowed to stand for 1 hour to be first coated.
  • the mixture After adding 12% collagen aqueous solution to 25% of the recovered cells and homogenizing the cells, the mixture was left to stand for 1 hour and then coated with 20% dextrin solution to 25% of the recovered cells. After homogenization, the mixture was allowed to stand for 1 hour and subjected to tertiary coating. In this case, the same cells without coating treatment were used as the control (sample 3) without any raw material.
  • the samples of Table 1 were prepared according to the method of Example 2, and then 4 weeks at 4, 25, and 40 ° C., respectively.
  • the viable cell counts were measured at two week intervals during storage.
  • the cells stored at each storage temperature were suspended in sterile saline solution, aliquoted and smeared according to dilution ratio in MRS (Difco) solid medium, and then cultured for 48 hours at 37 ° C. to measure colony numbers. .
  • the survival rate (%) is expressed as [(Log index value at Week 4 / Log index value at Week 0) X 100].
  • Experiment group 1 Control Week 0 4.6 X 10 8 (CFU / mg) 2.8 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 2.0 X 10 8 (CFU / mg) 2.4 X 10 8 (CFU / mg) 2.5 X 10 7 (CFU / mg) 4 Weeks 2.0 X 10 8 (CFU / mg) 2.2 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) Survival Rate after Week 4 (%) 95.8 (%) 98.8 (%) 92.6 (%)
  • Experiment group 1 Experiment group 2 Control Week 0 4.6 X 10 8 (CFU / mg) 2.8 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 8.3 X 10 7 (CFU / mg) 9.3 X 10 7 (CFU / mg) 1.3 X 10 6 (CFU / mg) 4 Weeks 8.9 X 10 7 (CFU / mg) 5.0 X 10 7 (CFU / mg) 5.1 X 10 6 (CFU / mg) Survival Rate after Week 4 (%) 91.8 (%) 91.2 (%) 82.9 (%)
  • Experiment group 1 Experiment group 2 Control Week 0 4.6 X 10 8 (CFU / mg) 2.8 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 1.6 X 10 7 (CFU / mg) 1.5 X 10 7 (CFU / mg) 1.7 X 10 3 (CFU / mg) 4 Weeks 4.5 X 10 6 (CFU / mg) 1.2 X 10 7 (CFU / mg) 4.5 X 10 3 (CFU / mg) Survival Rate after Week 4 (%) 76.8 (%) 83.9 (%) 45.1 (%)
  • the samples of Table 1 were prepared according to the method of Example 2 and then added with artificial gastric juice of pH 2.0 and 2.5, respectively, for 30 minutes, After reacting for 60 minutes and 120 minutes, the solution was diluted 10-fold with sterile saline solution and dispensed and plated according to the dilution ratio in MRS (Difco) solid medium. Thereafter, the colonies were measured by incubating at 37 ° C. for 48 hours, and the survival rate (%) was expressed as [(Log index value after initial gastric fluid exposure / initial Log index value) ⁇ 100].
  • the survival rate of the cells showed a tendency to decrease as the time of exposure to gastric juice (pH solution) increased, and coating treatment was performed at both pH 2.0 and pH 2.5 conditions. Compared with the control group did not show a high survival rate in the three-coated experiment groups 1 and 2. In addition, it was confirmed that the triple coating method significantly increased acid resistance compared to the case where no treatment was observed when maintaining a survival rate of about 95% or more in an acidic environment of 98% or more and pH 2.0 in a pH 2.5 condition.
  • Experiment group 1 Control 0 min 2.0 X 10 8 (CFU / mg) 2.1 X 10 8 (CFU / mg) 1.7 X 10 7 (CFU / mg) 30 minutes 2.8 X 10 8 (CFU / mg) 9.4 X 10 7 (CFU / mg) 2.1 X 10 7 (CFU / mg) 60 minutes 1.0 X 10 8 (CFU / mg) 1.1 X 10 8 (CFU / mg) 6.1 X 10 6 (CFU / mg) 120 minutes 7.9 X 10 7 (CFU / mg) 7.4 X 10 7 (CFU / mg) 4.8 X 10 6 (CFU / mg) Survival rate after 120 minutes (%) 95.0 (%) 94.5 (%) 92.5 (%)
  • Experiment group 1 Control 0 min 2.0 X 10 8 (CFU / mg) 2.1 X 10 8 (CFU / mg) 1.4 X 10 7 (CFU / mg) 30 minutes 2.3 X 10 8 (CFU / mg) 1.7 X 10 8 (CFU / mg) 1.4 X 10 7 (CFU / mg) 60 minutes 2.2 X 10 8 (CFU / mg) 1.7 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) 120 minutes 1.6 X 10 8 (CFU / mg) 1.6 X 10 8 (CFU / mg) 6.7 X 10 6 (CFU / mg) Survival rate after 120 minutes (%) 98.8 (%) 98.4 (%) 94.4 (%)
  • the samples of Table 1 were prepared according to the method of Example 2 and then bile solution (Bile extract 0.3%, 2.0%) The reaction was carried out for 30 minutes, 60 minutes and 120 minutes, and then diluted 10-fold with sterile saline, and then dispensed and plated according to the dilution ratio in MRS (Difco) solid medium. After culturing at 37 ° C. for 48 hours to determine colony numbers, the survival rate (%) was expressed as [(Log index value after initial bile acid exposure / initial Log index value) ⁇ 100].
  • Experiment group 1 Control 0 min 1.9 X 10 8 (CFU / mg) 1.4 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) 30 minutes 2.9 X 10 7 (CFU / mg) 3.6 X 10 7 (CFU / mg) 3.0 X 10 6 (CFU / mg) 60 minutes 3.3 X 10 7 (CFU / mg) 3.8 X 10 7 (CFU / mg) 5.8 X 10 6 (CFU / mg) 120 minutes 6.3 X 10 7 (CFU / mg) 3.7 X 10 7 (CFU / mg) 2.6 X 10 6 (CFU / mg) Survival rate after 120 minutes (%) 94.3 (%) 92.8 (%) 85.7 (%)
  • Experiment group 1 Control 0 min 1.9 X 10 8 (CFU / mg) 1.4 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) 30 minutes 2.3 X 10 7 (CFU / mg) 2.7 X 10 7 (CFU / mg) 4.0 X 10 6 (CFU / mg) 60 minutes 2.6 X 10 7 (CFU / mg) 3.1 X 10 7 (CFU / mg) 2.4 X 10 6 (CFU / mg) 120 minutes 4.1 X 10 7 (CFU / mg) 5.0 X 10 7 (CFU / mg) 3.3 X 10 6 (CFU / mg) Survival rate after 120 minutes (%) 92.0 (%) 94.4 (%) 87.1 (%)
  • the samples of Table 1 were prepared according to the method of Example 2 after digestion enzyme solution, that is, 0.2% pancreatin (pancreatin) and 100 unit / mL of alpha-amylase were added, respectively, and reacted for 30 minutes, 60 minutes, and 120 minutes, and then diluted 10-fold with sterile saline solution to dilute the MRS (Difco) solid medium. Dispensed and smeared accordingly. Colonies were measured after incubation at 37 ° C. for 48 hours. The survival rate (%) was expressed as [(Log index value / initial Log index value after digestive enzyme solution exposure) ⁇ 100].
  • Experiment group 1 Control 0 min 2.2 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2.8 X 10 7 (CFU / mg) 30 minutes 3.0 X 10 8 (CFU / mg) 2.2 X 10 8 (CFU / mg) 2.7 X 10 7 (CFU / mg) 60 minutes 3.5 X 10 8 (CFU / mg) 2.4 X 10 8 (CFU / mg) 3.2 X 10 7 (CFU / mg) 120 minutes 2.4 X 10 8 (CFU / mg) 2.0 X 10 8 (CFU / mg) 1.1 X 10 7 (CFU / mg) Survival rate after 120 minutes (%) 100.4 (%) 102.6 (%) 94.8 (%)
  • Experiment group 1 Control 0 min 1.5 X 10 8 (CFU / mg) 1.5 X 10 8 (CFU / mg) 2.2 X 10 7 (CFU / mg) 30 minutes 3.3 X 10 8 (CFU / mg) 2.4 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) 60 minutes 3.6 X 10 8 (CFU / mg) 2.5 X 10 8 (CFU / mg) 2.1 X 10 7 (CFU / mg) 120 minutes 2.9 X 10 8 (CFU / mg) 3.1 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) Survival rate after 120 minutes (%) 103.4 (%) 104.1 (%) 97.9 (%)
  • the samples of Table 1 were prepared according to the method of Example 2 and then the temperature at 40 °C, 50 °C, and 70 °C Sterile saline solution was added to each sample and maintained at the same temperature for 10 minutes and 30 minutes in a constant temperature bath, and then diluted 10 times with sterile saline solution and dispensed and plated according to the dilution ratio in MRS (Difco) solid medium. After 48 hours of incubation at 37 °C colony number was measured, the survival rate (%) is expressed as [(Log index value / initial Log index value after heat treatment) X 100].
  • the difference in thermal stability by coating treatment was more prominent at 70 °C high temperature condition, whereas the control group without coating was not identified after living treatment for 30 minutes.
  • the experimental group 2 which is greatly improved to the above and triple-coated, has great significance in that the cell survival rate is improved to 20% or more. Therefore, in view of the previous results, the triple-coated sample of Lactobacillus pentosus KF340 protects the strain from the external environment, thereby increasing the storage stability, acid resistance, bile resistance, stability of digestive enzymes, and thermal stability according to storage temperature. It was found to improve.
  • Experiment group 1 Control 0 min 1.4 X 10 8 (CFU / mg) 1.4 X 10 8 (CFU / mg) 1.9 X 10 7 (CFU / mg) 10 minutes 2.4 X 10 8 (CFU / mg) 2.1 X 10 8 (CFU / mg) 1.2 X 10 7 (CFU / mg) 30 minutes 2.2 X 10 8 (CFU / mg) 1.7 X 10 8 (CFU / mg) 1.0 X 10 7 (CFU / mg) Survival rate after 30 minutes (%) 102.4 (%) 101.2 (%) 96.4 (%)
  • Experiment group 1 Control 0 min 1.4 X 10 8 (CFU / mg) 1.4 X 10 8 (CFU / mg) 1.9 X 10 7 (CFU / mg) 10 minutes 4.8 X 10 7 (CFU / mg) 4.2 X 10 7 (CFU / mg) 3.6 X 10 6 (CFU / mg) 30 minutes 2.8 X 10 7 (CFU / mg) 2.2 X 10 7 (CFU / mg) 1.6 X 10 6 (CFU / mg) Survival rate after 30 minutes (%) 91.4 (%) 90.3 (%) 85.1 (%)
  • Experiment group 1 Control 0 min 2.1 X 10 8 (CFU / mg) 2.1 X 10 8 (CFU / mg) 1.1 X 10 7 (CFU / mg) 10 minutes 4.3 X 10 4 (CFU / mg) 9.0 X 10 4 (CFU / mg) 7.3 X 10 3 (CFU / mg) 30 minutes 5.5 X 10 2 (CFU / mg) 5.0 X 10 1 (CFU / mg) - Survival rate after 30 minutes (%) 33.0 (%) 20.4 (%) 0.0 (%)
  • the present invention is xylitol; Gelatin or collagen; And by sequentially coating the lactic acid bacteria with dextrin in triplicate, it was experimentally confirmed that the stability to storage temperature and heat, acid resistance, bile resistance, and digestive enzymes were improved more than lactic acid bacteria which were not conventionally coated.
  • the present invention can be maintained in the intestinal physiological activity through the gastrointestinal tract without killing the lactic acid bacteria enhanced stability by triple coating, fermented milk, fermented foods, functional foods, general foods, cosmetics, and pharmaceuticals It is expected to be usefully used in related industries.

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Abstract

La présente invention concerne des bactéries lactiques présentant une stabilité accrue, et leur procédé de production, plus particulièrement des bactéries lactiques présentant des propriétés améliorées de stabilité thermique et au stockage, de résistance aux acides, de résistance à la bile et de stabilité par rapport aux enzymes digestives grâce à un triple revêtement, dans l'ordre, de xylitol, de gélatine ou de collagène, et de dextrine et un procédé de production des bactéries lactiques. En appliquant un triple revêtement, dans l'ordre, de xylitol, de gélatine ou de collagène, et de dextrine, les bactéries lactiques produites au moyen du procédé de revêtement selon la présente invention présentent, par rapport aux bactéries lactiques classiques sans revêtement, une température de stockage supérieure et de meilleures propriétés confirmées expérimentalement de stabilité thermique, de résistance aux acides, de résistance à la bile et de stabilité par rapport aux enzymes digestives, et donc une résistance aux contraintes externes, une viabilité accrue de la souche lors du passage à travers le tractus gastro-intestinal, et la capacité à maintenir la bioactivité originale dans l'intestin ont été confirmées, et par conséquent le procédé de revêtement selon la présente invention devrait être appliqué efficacement dans des industries apparentées à l'aide de bactéries lactiques, telles que du lait fermenté, des aliments fermentés, des aliments fonctionnels, des aliments de grande consommation, des produits cosmétiques et des médicaments.
PCT/KR2017/014806 2016-12-16 2017-12-15 Bactéries lactiques présentant une stabilité accrue, et leur procédé de production Ceased WO2018111023A1 (fr)

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KR20160172787 2016-12-16
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KR1020170172396A KR102004204B1 (ko) 2016-12-16 2017-12-14 안정성이 증진된 유산균 및 이의 제조방법
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100387245B1 (ko) * 1997-10-17 2003-08-19 일양약품주식회사 유산균의안정화를위한미세장용성코팅과립
US7229818B2 (en) * 2003-01-14 2007-06-12 Randolph Stanley Porubcan Formulations to increase in vivo survival of probiotic bacteria and extend their shelf-life
KR20070104140A (ko) * 2006-04-21 2007-10-25 (주)케비젠 유산균 다중 마이크로캡슐의 제조방법, 이 방법에 의해제조된 마이크로캡슐 및 이를 포함하는 제품
KR20090082035A (ko) * 2008-01-25 2009-07-29 정명준 3중 코팅 유산균의 제조방법 및 나노 입자 코팅 방법, 그방법으로 제조된 3중 코팅 유산균 및 이를 포함하는 제품
US20150359894A1 (en) * 2006-09-27 2015-12-17 Little Calumet Holdings, Llc Probiotic oral dosage forms

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100387245B1 (ko) * 1997-10-17 2003-08-19 일양약품주식회사 유산균의안정화를위한미세장용성코팅과립
US7229818B2 (en) * 2003-01-14 2007-06-12 Randolph Stanley Porubcan Formulations to increase in vivo survival of probiotic bacteria and extend their shelf-life
KR20070104140A (ko) * 2006-04-21 2007-10-25 (주)케비젠 유산균 다중 마이크로캡슐의 제조방법, 이 방법에 의해제조된 마이크로캡슐 및 이를 포함하는 제품
US20150359894A1 (en) * 2006-09-27 2015-12-17 Little Calumet Holdings, Llc Probiotic oral dosage forms
KR20090082035A (ko) * 2008-01-25 2009-07-29 정명준 3중 코팅 유산균의 제조방법 및 나노 입자 코팅 방법, 그방법으로 제조된 3중 코팅 유산균 및 이를 포함하는 제품

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