WO2010101175A1 - Poudre lyophilisée de cellules microbiennes et son procédé de fabrication - Google Patents

Poudre lyophilisée de cellules microbiennes et son procédé de fabrication Download PDF

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Publication number
WO2010101175A1
WO2010101175A1 PCT/JP2010/053421 JP2010053421W WO2010101175A1 WO 2010101175 A1 WO2010101175 A1 WO 2010101175A1 JP 2010053421 W JP2010053421 W JP 2010053421W WO 2010101175 A1 WO2010101175 A1 WO 2010101175A1
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freeze
trehalose
sucrose
producing
dried powdered
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PCT/JP2010/053421
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English (en)
Japanese (ja)
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政幸 上條
正樹 寺原
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明治乳業株式会社
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Priority to JP2011502775A priority Critical patent/JP5583114B2/ja
Priority to SG2011049061A priority patent/SG172859A1/en
Priority to CN201080004172.6A priority patent/CN102272287B/zh
Publication of WO2010101175A1 publication Critical patent/WO2010101175A1/fr
Priority to HK12100228.0A priority patent/HK1159688A1/xx

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    • 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
    • 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
    • C12N1/04—Preserving or maintaining viable microorganisms

Definitions

  • the present invention relates to a freeze-dried powdered cell and a method for producing the same, and particularly to a freeze-dried powdered cell that can be stored for a long time even under a predetermined high temperature condition and a method for producing the same.
  • a large number of processed food groups containing these fungi have been developed for the purpose of improving and normalizing the digestive tract. Moreover, the device for ingesting these fungi simply and effectively is also made
  • Patent Document 1 by coating and granulating useful bacteria such as lactic acid bacteria and bifidobacteria that die under acidic or high temperature, the solubility in the intestine is not improved in the gastric juice in the sense of preservation. want to be.
  • a physiologically active substance is granulated by three-layer coating, and sugar is used as one of them.
  • Patent Document 2 proposes a method of adding a survival improver containing a saccharide selected from glycerol, xylitol, adonitol, arabitol, and mannitol to improve the survival of Bifidobacterium. Yes.
  • the purpose of this method is to maintain the survival and the number of bacteria during storage under aerobic conditions and low pH conditions.
  • this technique aims to improve the survival rate of the system in the medium.
  • Patent Document 3 a polyglycerol fatty acid ester is mixed with lactic acid bacteria cell powder to provide an enteric lactic acid bacteria composition having excellent long-term stability, acid resistance and enteric properties.
  • the above technique focuses on storage and tolerance under specific conditions.
  • Patent Document 4 discloses that a lot of konjac powder derived from lactic acid bacteria is dispersed in a lactic acid bacteria dispersion for the purpose of providing a production method with little damage or death of the bacteria in the freezing or freeze-drying process and having a high survival rate.
  • a method of freeze-drying by adding a saccharide partial hydrolyzate is disclosed.
  • the polysaccharide partial hydrolyzate has a predetermined molecular weight obtained by enzymatic degradation, and requires a complicated process for producing it.
  • this technique is mainly intended to improve the survival rate in freeze-thawing, and there is no suggestion or reference regarding the survival rate after storage of freeze-dried powdered cells produced by this technique.
  • Patent Document 5 proposes a preservation solution having a good survival rate and activity even after thawing frozen bacteria.
  • This cryopreservation solution for fungi contains trehalose and / or polyethylene glycol as active ingredients. This is characterized by the fact that it is a preservative solution that can be used immediately after thawing, while preventing damage during freezing. It is a system that contains water, and is a technology that solves the problem in the process of freezing to thawing.
  • Lactobacillus fermentum is coated with a protective film containing starch and saccharide to form a spray-dried powder.
  • This technique is a technique that focuses on enhancing the survival of bacteria during spray drying.
  • Patent Document 7 improves the storage stability in a dry state by allowing a dry microorganism such as a dry lactic acid bacterium to coexist with an L-arginine acidic amino acid salt.
  • This technology is mainly intended to improve the storage stability of microorganisms when formulating microorganisms (fungi).
  • the results of bifidobacteria preparations produced by this technique were stored for 2 weeks at 40 ° C., the survival rate is only about 25%, and it cannot withstand long-term storage for several months. It was.
  • Non-Patent Document 1 from the mechanism analysis in lyophilization, it was found that the damage during lyophilization of microorganisms is largely due to changes in the physical state of phospholipids constituting the cell membrane and structural changes in proteins, In contrast, the results of lyophilization of Escherichia coli and the like in the presence of trehalose or sucrose are shown. However, there is no suggestion or mention about using a mixture of trehalose and sucrose. Furthermore, no investigation has been made on the long-term storage of specific freeze-dried products and storage at a predetermined high temperature condition.
  • Non-Patent Document 2 studies on lyophilization and storage of Lactobacillus salivarius have been conducted for the same purpose as described above.
  • sucrose and 18% nonfat dry milk as a freeze-drying protective agent
  • freeze-dried powdered cells are stored at -85 ° C, and the survival rate is examined.
  • the sugar concentration in the cell suspension at the time of freeze-drying is about 3.2%
  • the nonfat dry milk concentration is 14.4%.
  • Non-Patent Document 2 shows the results of a storage test under environmental conditions that deviate from the normal living environment, and there is no suggestion or reference regarding storage under high temperature conditions. It was still difficult to apply the technology to general food.
  • powdery microbial cell for use as a powdery microbial cell that can be applied to foods that can be stored at room temperature for a long period of time, it can be used under special conditions at high temperatures (30-40 ° C) within the range expected for normal living environment temperatures. It is necessary to have high survivability without limitation of storage conditions. A powdery cell having no such property cannot guarantee its quality and cannot be used as a food material.
  • an object of the present invention is to provide a lyophilized powdery microbial cell suitable for long-term storage and obtained by simple and effective lyophilization, and a method for producing the same.
  • lactic acid bacteria and bifidobacteria are useful even if they are stored for a long time after freeze-drying, or even if they are stored in a living temperature range (especially in summer) that is considered to have a significant impact on storage stability, not at very low temperatures.
  • a lyophilized powdery cell that can be stored in a state where the survival rate of the fungus is high, can be directly ingested as a powder after storage, or can be effectively used as a raw material in food production, and its production A method is provided.
  • the food composition containing the obtained freeze-dried powdered microbial cell is also provided.
  • the present invention provides a lyophilized powdery microbial cell suitable for long-term storage and obtained by simple and effective lyophilization.
  • a lyophilized powdery cell is obtained by freeze-drying a suspension in which lactic acid bacteria and / or bifidobacteria are suspended in a saccharide solution to obtain a lyophilized powdered cell.
  • the concentration of each of trehalose and sucrose is preferably 4.5 to 15% by weight, more preferably 8 to 12% by weight.
  • the weight ratio of trehalose to sucrose is preferably 5: 1 to 1: 5.
  • preferred examples of the lactic acid bacteria and / or bifidobacteria include fungi belonging to the genus Bifidobacterium.
  • fungi belonging to the genus Bifidobacterium include Bifidobacterium bifidum strains, Bifidobacterium longum strains, and the like.
  • Bifidobacterium bifidum strain include the Bifidobacterium bifidum OLB6378 strain.
  • a preferred example of the Bifidobacterium longum strain is a Bifidobacterium longum OLB6001 strain.
  • Another aspect of the present invention is a freeze-dried powdered cell produced by the method for producing a freeze-dried powdered cell as described above.
  • Still another embodiment of the present invention is a food composition comprising an effective amount of the above lyophilized powdered microbial cells.
  • the method for producing a freeze-dried powdered cell that can be stored for a long period of time according to the present invention is particularly effective when it is necessary to guarantee long-term storage, such as a solid or powdered food. That is, the powder freeze-dried powder produced by the production method of the present invention has a significantly improved survival rate during high-temperature storage, so that it is useful even after use after storage, such as lactic acid bacteria and / or bifidobacteria. The effect of can be obtained.
  • the production method of the present invention is a simple method, it does not require any special equipment or complicated steps, and there is no increase in costs associated therewith.
  • the lyophilized powdery cell produced by the production method of the present invention can be used as it is, it can be easily ingested directly after storage or effectively used as a raw material in food production. be able to.
  • the trehalose concentration and the sucrose concentration in the suspension before lyophilization were each 6.6% by weight (a sugar solution containing 20% by weight of trehalose and sucrose was used as a sugar solution before mixing with the cells). It is a graph which shows the result of the storage test in 20 degreeC, 30 degreeC, and each temperature of 40 degreeC of the freeze-dried powdery cell body of Bifidobacterium bifidum prepared on a certain condition.
  • the conditions in which the trehalose concentration and the sucrose concentration in the suspension before lyophilization are 10% by weight (a solution containing 30% by weight of trehalose and sucrose as a sugar solution before mixing with the cells) were used.
  • Examples of lactic acid bacteria and bifidobacteria used in the present invention include bacteria belonging to the genus Bifidobacterium, such as Bifidobacterium longum strains, Bifidobacterium infantitis (Bifidobacterium infantitis). ) Strains, Bifidobacterium breve strains, Bifidobacterium bifidum strains, Bifidobacterium adrescentis strains and the like. Still further, fungi belonging to the genus Lactobacillus and Streptococcus can be exemplified, for example, Lactobacillus gasseri strain, Lactobacillus bulgaricus strain, Etc. can be specifically mentioned. However, the present invention is not limited to these species, and these strains can be used alone or in combination of two or more.
  • preferable examples include a Bifidobacterium bifidum strain and a Bifidobacterium longum strain.
  • Bifidobacterium bifidum OLB6378 strain, Bifidobacterium longum OLB6001 strain and the like are mentioned as examples of deposited strains of these preferable examples.
  • (A) Deposit of Bifidobacterium bifidum OLB6378 strain The Bifidobacterium bifidum OLB6378 strain used in the present invention was deposited under the following conditions. (1) Depositary Institution: National Institute of Technology and Evaluation, Patent Microorganisms Deposit Center (2) Contact: 2-5-8 Kazusa Kamashichi, Kisarazu City, Chiba Prefecture 292-0818 Phone number 0438-20-5580 (3) Accession number: NITE BP-31 (4) Display for identification: Bifidobacterium bifidum OLB6378 (5) Original deposit date: October 26, 2004 (6) Date of transfer to deposit under the Budapest Treaty: January 18, 2006 (B) Deposit of Bifidobacterium longum OLB6001 strain The Bifidobacterium longum OLB6001 strain used in the present invention was deposited under the following conditions.
  • the Bifidobacterium bifidum OLB6378 strain and the Bifidobacterium longum OLB6001 strain used in the present invention have the following mycological properties.
  • Bifidobacterium bifidum OLB6378 is a Gram-positive obligate anaerobe derived from human infant feces. Lactobacilli MRS Broth (BD) is inoculated with this bacterium, and cultured in anaerobic state by using AnaeroPack Kenki (manufactured by Mitsubishi Gas Chemical Company) at 37 ° C. for 18 hours, a Y-shaped microbial form is observed.
  • BiBIF- a specific primer for Bifidobacterium bifidum (intestinal flora symposium 8, molecular biological detection / identification of intestinal flora, Tomohiro Mitsuoka, Takahiro Matsumoto), specifically, BiBIF-, which is a species-specific primer for 16S rDNA.
  • PCR products were observed by PCR using 1: CCA CAT GAT CGC ATG TGA TT and BiBIF-2: CCG AAG GCT TGC TCC CAA A.
  • the Bifidobacterium longum OLB6001 strain is a Gram-positive obligate anaerobic bacterium derived from human adult feces, and the shape of the fungus is a gonococcal or branched polymorph and does not have spore formation or motility.
  • This bacterium is applied on a BL agar medium (Eiken) plate and cultured by a steel wool method at 37 ° C. for 48 hours, colonies having an opaque circular hemispherical gloss are formed.
  • the saccharide as a protective agent used in the present invention is a mixture of trehalose and sucrose.
  • the concentration of trehalose and sucrose in the suspension before lyophilization is 4.5% by weight or more, preferably 8% by weight, from the viewpoint of obtaining lyophilized powdery cells suitable for long-term storage at high temperatures. That's it.
  • the upper limit of the concentration is preferably 15% by weight, more preferably 12% by weight from the viewpoint that the effect of long-term storage stability at high temperatures is equivalent even if the concentration is excessively increased.
  • the weight ratio of trehalose to sucrose is not particularly limited, but is preferably 5: 1 to 1: 5, more preferably 3: 1 from the viewpoint of obtaining lyophilized powder cells suitable for long-term storage at high temperatures.
  • the carbohydrates are preferably mixed with cultured bacteria in the form of a carbohydrate-containing aqueous solution, and the bacteria are resuspended.
  • the lyophilized powdery cell of the present invention can be obtained by freeze-drying the suspension thus obtained.
  • the saccharide-containing aqueous solution may contain, for example, milk protein, amino acids, ascorbic acid and the like in addition to water and saccharides.
  • the manufacturing method of the lyophilized powdery microbial cell of this invention is not specifically limited, For example, it consists of the following procedures. 1) A desired fungus is cultured according to a conventional method. 2) Use the culture solution containing the cultured bacteria as it is in the next 3), or concentrate or solid-liquid separate the culture solution containing the cultured bacteria by centrifugation or the like. Bacteria separated as a liquid (bacterial cell liquid) or solid content is obtained and used in the following 3). 3) The obtained bacterial cell liquid or bacterial cell and a solution containing a predetermined concentration of sugar (protective agent) are mixed to obtain a suspension, and then the suspension is freeze-dried to obtain the present invention. To obtain a freeze-dried powdered cell.
  • freeze-drying method examples include a method using a freeze-dryer. For example, after rapid pre-freezing at a low temperature (for example, ⁇ 30 to ⁇ 90 ° C.), the temperature is reduced at room temperature (for example, 0 to 20 ° C.). It may be dried under a low pressure (preferably 1000 Pa or less, more preferably 100 Pa or less), and the temperature of the freeze dryer is increased to, for example, 30 to 70 ° C. while maintaining the degree of vacuum. .
  • the lyophilized powdery cell of the present invention can be ingested as it is because it uses trehalose and sucrose which can also be used as food additives, and it can be added to various food compositions. Can also be used.
  • the freeze-dried powdered cell of the present invention shows a predetermined survival rate even after long-term storage, it can be used as a raw material for inoculum such as fermented milk.
  • the food composition examples include various foods and beverages (soft drinks, fermented milk, yogurt, adjusted powdered milk, etc.).
  • the food composition can be used as it is, or can be used according to a conventional method for ordinary food compositions such as mixing with other foods or food ingredients.
  • the food composition may be in the state of a commonly used food or drink, for example, solid (powder, granule, etc.), paste, liquid or suspension.
  • components in the food composition are not particularly limited, and examples include water, protein, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, and flavors. These components may be used individually by 1 type, or can also be used in combination of 2 or more type. In addition, as another component in a food composition, you may use the foodstuff which contains many synthetic products and / or synthetic products.
  • Example 1 According to the procedure shown below, freeze-dried powdered cells were produced based on the production method of the present invention. 1) The Bifidobacterium bifidum OLB6378 strain was subjected to neutralization culture in a casein decomposition medium (a medium containing enzymatically decomposed casein as a protein). 2) Centrifugation (320 ° C., 10000 G for 20 minutes) of 320 ml of the culture solution was performed to remove 307.2 ml of the supernatant to obtain a cell pellet fraction (12.8 ml).
  • casein decomposition medium a medium containing enzymatically decomposed casein as a protein
  • the protective agent liquid used in this experimental example is as shown in Table 1 below.
  • Table 2 shows the results of the Bifidobacterium bifidum preservation test for Samples 1 and 2 and Comparative Samples 1, 2 and 3 according to the present invention.
  • the survival rate was low about the comparative samples 1 and 2
  • saving for 7 days at 20 degreeC is described together for reference.
  • Comparative Examples 1 and 2 are tests on a trehalose-only system typified by Non-Patent Document 2, but it was found that the objective could not be sufficiently achieved with trehalose alone.
  • Non-Patent Document 2 describes the effect of storage under ultra-low temperature and humidity conditions, but it has been found that the desired effect cannot be obtained at the living environment temperature as in this experiment.
  • Example 2 Experiments were conducted on the effects of long-term storage (up to 6 months) of the freeze-dried microbial cells obtained by the production method of the present invention under high temperature conditions (20 ° C, 30 ° C, 40 ° C). .
  • the freeze-dried powdered cells used were produced according to the same production method as Samples 1 and 2, except that the amount charged was increased about 30 times. Therefore, it demonstrates as the samples 1 and 2 below.
  • the results obtained by the above experiment are shown in FIGS.
  • sample 1 (see FIG. 1), containing 6.6% by weight of each of trehalose and sucrose obtained by mixing a solution containing 20% by weight of trehalose and a solution containing 20% by weight of sucrose.
  • Protectant solution and sample 2 (see FIG. 2), each containing 10% by weight of trehalose and sucrose obtained by mixing a solution containing 30% by weight trehalose and a solution containing 30% by weight sucrose.
  • Both the protective agent solution maintained a sufficient viable count even after 6 months even under storage temperature conditions of 20 ° C. and 30 ° C., and showed very high survival. Although it is a considerably extreme living environment temperature condition, with respect to storage at 40 ° C., the sample 1 shows 93.9% survival after 1 month storage. Therefore, it was found that the concentration used in Sample 1 was more preferable as the concentration of the protective agent solution under these conditions.
  • Example 3 Experiments were performed in the same manner as in Example 1 except that the samples shown in Table 3 below were used and the samples were stored at 40 ° C. for 3 months. The results are shown in Table 4. From Table 4, it was found that there was no difference between the mass ratio of trehalose and sucrose in the range of 3: 1 to 1: 3 and the survival rate was 11 to 17%.
  • Example 4 According to the procedure shown below, freeze-dried powdered cells were produced based on the production method according to the present invention.
  • Bifidobacterium longum OLB6001 strain was neutralized and cultured in a casein decomposition medium (a medium containing enzymatically decomposed casein as a protein).
  • a casein decomposition medium a medium containing enzymatically decomposed casein as a protein.
  • Centrifugation 5 ° C., 10000 G for 20 minutes
  • 5400 ml of the culture broth removed 5200 ml of the supernatant to obtain a cell pellet fraction (200 ml).
  • 3) 100 ml of a protective agent solution containing a predetermined concentration of sugar was added to the cell pellet fraction (200 ml), the cells were suspended, frozen at ⁇ 80 ° C.
  • freeze-dried To 1 g of freeze-dried powdered cells immediately after lyophilization, physiological saline was added to cover the cells, and the number of viable bacteria in the water-covered solution was measured using a BL agar plate medium. 5) Furthermore, freeze-dried after 2 g of freeze-dried powdered cells were placed in lamizip (registered trademark; trade name of plastic bag) and stored at 40 ° C. for 8 days, 30 days, 82 days, and 124 days. Similarly, the number of viable cells was measured on a BL agar plate medium for powdered cells.
  • lamizip registered trademark; trade name of plastic bag
  • Example 8 A sugar solution (hereinafter referred to as Sample 6) containing 20% by weight of trehalose and sucrose was used. The contents of trehalose and sucrose in the suspension before lyophilization formed by mixing the sugar solution and the cell pellet fraction were 6.7% by weight, respectively.
  • Comparative Example 5 A solution containing 6% by weight of skim milk powder, 1.7% by weight of lactose, 0.4% by weight of amino acid (lysine, etc.) and 4% by weight of other components (dextrin, etc.) (hereinafter referred to as Comparative Sample 4) is used. It was. The results are shown in Table 3 and FIG.
  • Table 5 shows that the freeze-dried microbial cell of Comparative Example 5 (circled number 1 in FIG. 3) shows only 0.002% survival when stored at 40 ° C. for about 4 months.
  • the freeze-dried microbial cell of Example 8 (circled number 2 in FIG. 3) shows a high survival rate of 40.0% when stored at 40 ° C. for about 4 months.
  • the method for producing lyophilized powdered cells that can be stored for a long period of time according to the present invention, lactic acid bacteria and / or useful after storage, especially when it is necessary to guarantee long-term storage, such as solids and powdered foods
  • the effect of bifidobacteria can be obtained.
  • no special equipment or complicated processes are required, and there is no cost increase associated therewith, which is economically advantageous.
  • the lyophilized powdery cell of the present invention can be used as it is, it can be easily ingested directly as a powder even after storage, or can be effectively used as a raw material in food production. Value is high.

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Abstract

L'invention porte sur une poudre lyophilisée de cellules microbiennes appropriée pour un stockage prolongé, qui peut être obtenue par un procédé de lyophilisation commode et efficace. L'invention porte sur un procédé de production d'une poudre lyophilisée de cellules microbiennes comprenant la lyophilisation d'une suspension dans laquelle des cellules d'une bactérie à acide lactique et/ou une bifidobactérie sont mises en suspension dans une solution de sucres, pour donner une poudre lyophilisée de cellules microbiennes, les sucres étant le tréhalose et le sucrose et les concentrations en tréhalose et sucrose dans la suspension avant la lyophilisation étant chacune supérieures ou égales à 4,5 % en poids. Le rapport en poids du tréhalose au sucrose est, de préférence, de 5:1 à 1:5.
PCT/JP2010/053421 2009-03-04 2010-03-03 Poudre lyophilisée de cellules microbiennes et son procédé de fabrication WO2010101175A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2011502775A JP5583114B2 (ja) 2009-03-04 2010-03-03 凍結乾燥粉末状菌体及びその製造方法
SG2011049061A SG172859A1 (en) 2009-03-04 2010-03-03 Freeze-dried microbial cell powder and method for producing same
CN201080004172.6A CN102272287B (zh) 2009-03-04 2010-03-03 冷冻干燥粉末状菌体及其制造方法
HK12100228.0A HK1159688A1 (en) 2009-03-04 2012-01-09 Freeze-dried microbial cell powder and method for producing same

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JP2009051118 2009-03-04
JP2009-051118 2009-03-04

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JP2012055288A (ja) * 2010-09-13 2012-03-22 Kaneka Corp 安定化された生菌製剤およびその製造方法。
JP2014068545A (ja) * 2012-09-27 2014-04-21 Meiji Co Ltd ビフィズス菌の検出方法
JP2014171423A (ja) * 2013-03-08 2014-09-22 Nissin Foods Holdings Co Ltd 凍結乾燥菌試料およびその製造方法
WO2016103699A1 (fr) * 2014-12-26 2016-06-30 株式会社明治 Promoteur de production d'acides organiques, et agent pour prévenir et/ou améliorer une maladie intestinale inflammatoire
JP2018007639A (ja) * 2016-07-15 2018-01-18 株式会社明治 微生物培養液の濃縮方法及び微生物濃縮物
JP2018526973A (ja) * 2016-07-15 2018-09-20 シージェイ チェイルジェダン コーポレーションＣｊ Ｃｈｅｉｌｊｅｄａｎｇ Ｃｏｒｐｏｒａｔｉｏｎ ガス発生量の低いリゥコノストック・メゼンテロイデスｃｊｌｍ１１９菌株およびこれを用いたキムチの製造方法
JP2019524097A (ja) * 2016-07-15 2019-09-05 シージェイ チェイルジェダン コーポレーションＣｊ Ｃｈｅｉｌｊｅｄａｎｇ Ｃｏｒｐｏｒａｔｉｏｎ ガス発生量の低いリゥコノストック・メゼンテロイデスｃｊｌｍ１８１菌株およびこれを用いたキムチの製造方法
JP2019525752A (ja) * 2016-07-15 2019-09-12 シージェイ チェイルジェダン コーポレーションＣｊ Ｃｈｅｉｌｊｅｄａｎｇ Ｃｏｒｐｏｒａｔｉｏｎ ガス発生量の低いリゥコノストック・メゼンテロイデスｃｊｌｍ６２７菌株およびこれを用いたキムチの製造方法
CN111534434A (zh) * 2020-06-28 2020-08-14 江南大学 一种冻干保护剂及其在冻干青春双歧杆菌中的应用

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JP2019524097A (ja) * 2016-07-15 2019-09-05 シージェイ チェイルジェダン コーポレーションＣｊ Ｃｈｅｉｌｊｅｄａｎｇ Ｃｏｒｐｏｒａｔｉｏｎ ガス発生量の低いリゥコノストック・メゼンテロイデスｃｊｌｍ１８１菌株およびこれを用いたキムチの製造方法
JP2019525752A (ja) * 2016-07-15 2019-09-12 シージェイ チェイルジェダン コーポレーションＣｊ Ｃｈｅｉｌｊｅｄａｎｇ Ｃｏｒｐｏｒａｔｉｏｎ ガス発生量の低いリゥコノストック・メゼンテロイデスｃｊｌｍ６２７菌株およびこれを用いたキムチの製造方法
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