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WO2018111024A1 - 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
WO2018111024A1
WO2018111024A1 PCT/KR2017/014807 KR2017014807W WO2018111024A1 WO 2018111024 A1 WO2018111024 A1 WO 2018111024A1 KR 2017014807 W KR2017014807 W KR 2017014807W WO 2018111024 A1 WO2018111024 A1 WO 2018111024A1
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Prior art keywords
lactic acid
acid bacteria
coating
cfu
parts
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Korean (ko)
Inventor
엄기안
연성흠
손락호
박채리
필감방
오명환
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Huons Co Ltd
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Huons Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
    • A23B2/00—Preservation of foods or foodstuffs, in general
    • A23B2/70—Preservation of foods or foodstuffs, in general by treatment with chemicals
    • A23B2/725—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of liquids or solids
    • 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
    • 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
    • A23P20/10—Coating with edible coatings, e.g. with oils or fats
    • 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 with enhanced stability and a method for preparing the same, more specifically dextrin; Gelatin or collagen; And it relates to a lactic acid bacterium and a method for producing the same that is enhanced by storage and thermal stability, acid resistance, bile resistance, and digestive enzymes by triple coating sequentially xylitol.
  • 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 the 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 of lactic acid bacteria and stability in the body, and as a result, the lactic acid bacteria, preferably Lactobacillus pentosus strain, dextrin; Gelatin or collagen; And triple coating with sequentially using xylitol was confirmed that the storage stability, thermal stability, acid resistance, bile resistance, and stability to the digestive enzymes of the strain is improved, based on this, the present invention was completed.
  • the lactic acid bacteria preferably Lactobacillus pentosus strain, dextrin; Gelatin or collagen
  • triple coating with sequentially using xylitol was confirmed that the storage stability, thermal stability, acid resistance, bile resistance, and stability to the digestive enzymes of the strain is improved, based on this, the present invention was completed.
  • 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.
  • the dextrin in step (a) may be 1% to 30% concentration.
  • the gelatin or collagen in step (b) may be 1% to 30% concentration.
  • the xylitol in step (c) may be 10% to 50% concentration.
  • the lactic acid bacteria is Lactobacillus sp. , Genus Bifidobacterium ( Bifidobacterium) sp . ), Genus Streptococcus sp. , Genus Lactococcus sp . ) Enterococcus sp . ), Pediococcus sp . , Genus Leuconostoc sp .) genus, and Weissella sp. genus.
  • the lactic acid bacteria may be Lactobacillus pentosus .
  • the dextrin in the step (a), is the first coating step by mixing 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 xylitol may include 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.
  • Lactobacillus prepared by the coating method according to the invention is dextrin; Gelatin or collagen; And triple-coating sequentially using xylitol to experimentally confirm that the storage temperature and heat stability, acid resistance, bile resistance, and digestive enzyme stability are improved compared to the 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.
  • Figure 1 is a photograph of the final raw material after freeze-drying the Lactobacillus pentosus KF340 strain sequentially coated with dextrin, gelatin, and xylitol in triple.
  • Figure 2 is a photograph of the final raw material after lyophilizing Lactobacillus pentosus KF340 strain sequentially coated with dextrin, collagen, and xylitol in triple.
  • Figure 3 is a photograph taken with a scanning electron microscope of the Lactobacillus pentosus KF340 strain sequentially coated with dextrin, gelatin, and xylitol in triple.
  • Figure 4 is a photograph of the Lactobacillus pentosus KF340 strain sequentially coated with dextrin, collagen, and xylitol by scanning electron microscope.
  • 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 was completed by developing a coating method.
  • the present invention (a) adding a dextrin (dextrin) 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 xylitol to the secondary coated lactic acid bacterium and homogenizing the third coating to improve the stability of the lactic acid bacterium.
  • the present invention (a) adding a dextrin (dextrin) 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 xylitol to the secondary coated lactic acid bacterium 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.
  • the dextrin may be dissolved in distilled water at a concentration of 1% to 30%, preferably 1% to 15%, more preferably 3% to 7%, even more preferably 5%, and used. Pre-sterilized, cooled to room temperature and available for coating.
  • the dextrin is homogenized after adding 10 parts by weight to 100 parts by weight, preferably 50 parts by weight to 100 parts by weight, and more preferably 100 parts by weight which is the same amount as the recovered cells, based on 100 parts by weight of the cells recovered after the culture.
  • the cells can be first coated by standing at room temperature for 0.5 to 3 hours, more preferably 1 hour 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.
  • 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.
  • xylitol is dissolved in distilled water and may be in 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 xylitol 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 recovered cells after culturing and homogenized at room temperature.
  • 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 .) genus, and Weissella sp.
  • genus may be one or more strains, more preferably Lactobacillus pentosus ( Lactobacillus pentosus ), more preferably Lactobacillus pento Sus KF340 (Accession No. 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 dissolved each raw material in distilled water to a concentration of 5% dextrin, 6% gelatin, and 20% xylitol. Sterilized at 121 ° C. for 15 minutes and then cooled at room temperature. Next, the same amount of 5% dextrin aqueous solution was added to the recovered cells, the cells were homogenized, and left to stand for 1 hour, followed by primary coating. 6% gelatin aqueous solution was added to 25% of the recovered cells, homogenized, and left to rest for 1 hour, followed by secondary coating, followed by adding 20% xylitol solution to 25% of the recovered cells.
  • each raw material was dissolved in distilled water to a concentration of 5% dextrin, 12% collagen, and 20% xylitol, sterilized at 121 ° C. for 15 minutes, and then cooled at room temperature.
  • the same amount of 5% dextrin aqueous solution was added to the recovered cells, the cells were homogenized, and left to stand for 1 hour, followed by primary coating.
  • 12% collagen aqueous solution was added to 25% of the recovered cells, homogenized, and left to rest for 1 hour, followed by secondary coating, followed by adding 20% xylitol solution to 25% of the recovered cells.
  • 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. conditions, 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 Experiment group 2 Control Week 0 3.9 X 10 8 (CFU / mg) 4.0 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 1.9 X 10 8 (CFU / mg) 3.0 X 10 8 (CFU / mg) 2.5 X 10 7 (CFU / mg) 4 Weeks 2.2 X 10 8 (CFU / mg) 2.9 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) Survival Rate after Week 4 (%) 97.1 (%) 98.4 (%) 92.6 (%)
  • Experiment group 1 Experiment group 2 Control Week 0 3.9 X 10 8 (CFU / mg) 4.0 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 9.9 X 10 7 (CFU / mg) 9.7 X 10 7 (CFU / mg) 1.3 X 10 6 (CFU / mg) 4 Weeks 4.6 X 10 7 (CFU / mg) 8.3 X 10 7 (CFU / mg) 5.1 X 10 6 (CFU / mg) Survival Rate after Week 4 (%) 89.2 (%) 92.1 (%) 82.9 (%)
  • Experiment group 1 Experiment group 2 Control Week 0 3.9 X 10 8 (CFU / mg) 4.0 X 10 8 (CFU / mg) 1.2 X 10 8 (CFU / mg) 2 weeks 1.1 X 10 7 (CFU / mg) 5.4 X 10 6 (CFU / mg) 1.7 X 10 3 (CFU / mg) 4 Weeks 5.5 X 10 6 (CFU / mg) 4.5 X 10 6 (CFU / mg) 4.5 X 10 3 (CFU / mg) Survival Rate after Week 4 (%) 78.5 (%) 77.3 (%) 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. After that, the colonies were measured by incubating for 48 hours at 37 ° C., 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 94% or more even in an acidic environment of 98% or more at pH 2.5 and pH 2.0.
  • Experiment group 1 Experiment group 2 Control 0 min 1.4 X 10 8 (CFU / mg) 2.3 X 10 8 (CFU / mg) 1.7 X 10 7 (CFU / mg) 30 minutes 1.3 X 10 8 (CFU / mg) 1.8 X 10 8 (CFU / mg) 2.1 X 10 7 (CFU / mg) 60 minutes 1.9 X 10 8 (CFU / mg) 1.9 X 10 8 (CFU / mg) 6.1 X 10 6 (CFU / mg) 120 minutes 9.2 X 10 7 (CFU / mg) 8.3 X 10 7 (CFU / mg) 4.8 X 10 6 (CFU / mg) % Survival After 120 Minutes 97.8 (%) 94.6 (%) 92.5 (%)
  • Experiment group 1 Experiment group 2 Control 0 min 1.4 X 10 8 (CFU / mg) 2.3 X 10 8 (CFU / mg) 1.4 X 10 7 (CFU / mg) 30 minutes 1.7 X 10 8 (CFU / mg) 3.2 X 10 8 (CFU / mg) 1.4 X 10 7 (CFU / mg) 60 minutes 2.0 X 10 8 (CFU / mg) 1.8 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) 120 minutes 1.7 X 10 8 (CFU / mg) 1.9 X 10 8 (CFU / mg) 6.7 X 10 6 (CFU / mg) % Survival After 120 Minutes 101.2 (%) 98.8 (%) 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 measure 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 1.6 X 10 8 (CFU / mg) 2.0 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) 30 minutes 1.2 X 10 8 (CFU / mg) 5.5 X 10 7 (CFU / mg) 3.0 X 10 6 (CFU / mg) 60 minutes 3.1 X 10 7 (CFU / mg) 2.4 X 10 7 (CFU / mg) 5.8 X 10 6 (CFU / mg) 120 minutes 6.3 X 10 7 (CFU / mg) 3.8 X 10 7 (CFU / mg) 2.6 X 10 6 (CFU / mg) % Survival After 120 Minutes 95.1 (%) 91.4 (%) 85.7 (%)
  • Experiment group 1 Control 0 min 1.6 X 10 8 (CFU / mg) 2.0 X 10 8 (CFU / mg) 3.1 X 10 7 (CFU / mg) 30 minutes 2.3 X 10 7 (CFU / mg) 2.1 X 10 7 (CFU / mg) 4.0 X 10 6 (CFU / mg) 60 minutes 2.1 X 10 7 (CFU / mg) 4.1 X 10 7 (CFU / mg) 2.4 X 10 6 (CFU / mg) 120 minutes 2.1 X 10 7 (CFU / mg) 8.2 X 10 7 (CFU / mg) 3.3 X 10 6 (CFU / mg) % Survival After 120 Minutes 89.3 (%) 95.4 (%) 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, and diluted in 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) 2.1 X 10 8 (CFU / mg) 2.8 X 10 7 (CFU / mg) 30 minutes 2.3 X 10 8 (CFU / mg) 2.5 X 10 8 (CFU / mg) 2.7 X 10 7 (CFU / mg) 60 minutes 2.7 X 10 8 (CFU / mg) 3.2 X 10 8 (CFU / mg) 3.2 X 10 7 (CFU / mg) 120 minutes 2.3 X 10 8 (CFU / mg) 2.0 X 10 8 (CFU / mg) 1.1 X 10 7 (CFU / mg) % Survival After 120 Minutes 100.3 (%) 99.9 (%) 94.8 (%)
  • Experiment group 1 Control 0 min 2.8 X 10 8 (CFU / mg) 1.9 X 10 8 (CFU / mg) 2.2 X 10 7 (CFU / mg) 30 minutes 3.0 X 10 8 (CFU / mg) 3.1 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) 60 minutes 2.9 X 10 8 (CFU / mg) 3.6 X 10 8 (CFU / mg) 2.1 X 10 7 (CFU / mg) 120 minutes 2.7 X 10 8 (CFU / mg) 4.0 X 10 8 (CFU / mg) 1.5 X 10 7 (CFU / mg) % Survival After 120 Minutes 99.8 (%) 104.0 (%) 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 incubation for 48 hours at 37 °C colony number was measured, where the survival rate (%) was expressed as [(Log index value / initial Log index value after heat treatment) X 100].
  • the triple coated sample of Lactobacillus pentosus KF340 protects the strains from the external environment, so that storage stability, acid resistance, bile resistance, stability against digestive enzymes, and thermal stability according to storage temperature Overall improvement was found.
  • Experiment group 1 Experiment group 2 Control 0 min 8.4 X 10 7 (CFU / mg) 1.8 X 10 8 (CFU / mg) 1.9 X 10 7 (CFU / mg) 10 minutes 2.1 X 10 8 (CFU / mg) 2.4 X 10 8 (CFU / mg) 1.2 X 10 7 (CFU / mg) 30 minutes 2.0 X 10 8 (CFU / mg) 2.1 X 10 8 (CFU / mg) 1.0 X 10 7 (CFU / mg) % Survival after 30 minutes 104.7 (%) 100.9 (%) 96.4 (%)
  • Experiment group 1 Experiment group 2 Control 0 min 8.4 X 10 7 (CFU / mg) 1.8 X 10 8 (CFU / mg) 1.9 X 10 7 (CFU / mg) 10 minutes 4.2 X 10 7 (CFU / mg) 1.3 X 10 8 (CFU / mg) 3.6 X 10 6 (CFU / mg) 30 minutes 2.2 X 10 7 (CFU / mg) 1.8 X 10 7 (CFU / mg) 1.6 X 10 6 (CFU / mg) % Survival after 30 minutes 92.7 (%) 87.9 (%) 85.1 (%)
  • Experiment group 1 Control 0 min 1.6 X 10 8 (CFU / mg) 1.6 X 10 8 (CFU / mg) 1.1 X 10 7 (CFU / mg) 10 minutes 5.5 X 10 3 (CFU / mg) 4.0 X 10 3 (CFU / mg) 7.3 X 10 3 (CFU / mg) 30 minutes 5.0 X 10 1 (CFU / mg) 1.7 X 10 3 (CFU / mg) - % Survival after 30 minutes 20.7 (%) 39.2 (%) 0.0 (%)
  • the present invention is dextrin; Gelatin or collagen; And by sequentially coating the lactic acid bacteria three times using xylitol it was experimentally confirmed that the stability to storage temperature and heat, acid resistance, bile resistance, and digestive enzymes than the conventional lactic acid bacteria is improved.
  • 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 dextrine, de gélatine ou de collagène, et de xylitol, et un procédé de production des bactéries lactiques. En appliquant un triple revêtement, dans l'ordre, de dextrine, de gélatine ou de collagène, et de xylitol, les bactéries lactiques produites au moyen du procédé de revêtement selon la présente invention possèdent, par comparaison 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 de résistance aux contraintes externes, de viabilité accrue de la souche lors du passage à travers le tractus gastro-intestinal, et la capacité à maintenir la bioactivité d'origine 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 utilisant des bactéries lactiques, telles que celles liées au lait fermenté, aux aliments fermentés, aux aliments fonctionnels, aux aliments de grande consommation, aux produits cosmétiques et aux médicaments.
PCT/KR2017/014807 2016-12-16 2017-12-15 Bactéries lactiques présentant une stabilité accrue, et leur procédé de production Ceased WO2018111024A1 (fr)

Applications Claiming Priority (6)

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KR20160172803 2016-12-16
KR20160172818 2016-12-16
KR10-2016-0172818 2016-12-16
KR10-2016-0172803 2016-12-16
KR1020170172397A KR102048690B1 (ko) 2016-12-16 2017-12-14 안정성이 증진된 유산균 및 이의 제조방법
KR10-2017-0172397 2017-12-14

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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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