JP7444949B2 - Medium for lactic acid bacteria - Google Patents

Description

The present invention relates to a medium for lactic acid bacteria suitable for culturing lactic acid bacteria.

Until now, as a medium used for culturing lactic acid bacteria, MRS (deMan, Rogosa, Sharpe) medium has been widely used because it promotes the growth of all lactic acid bacteria.

There is no problem in culturing lactic acid bacteria in this MRS medium and measuring the number of bacteria or physical properties, but since MRS medium contains reagents that cannot be used as food materials, cultured lactic acid bacteria can be directly used. It could not be used for food purposes.

Until now, as a culture medium for lactic acid bacteria made of food materials, a culture medium made of barley shochu distillation residue to which sugars and emulsifiers have been added has been reported (Patent Document 1). There was a problem in that the culture medium did not reach a sufficient number of lactic acid bacteria.

Patent No. 4921499

An object of the present invention is to provide a medium for lactic acid bacteria that is made of a food material and can reach a sufficient number of bacteria by culturing the lactic acid bacteria, and a technique using the same.

As a result of intensive research to solve the above problems, the present inventors found that a medium containing soybean peptide and yeast extract can reach a sufficient number of bacteria by culturing lactic acid bacteria, and completed the present invention. .

That is, the present invention is a medium for lactic acid bacteria containing soybean peptide and yeast extract.

Further, the present invention is a method for culturing lactic acid bacteria, which comprises culturing lactic acid bacteria in the above-mentioned medium for lactic acid bacteria.

Furthermore, the present invention is a lactic acid bacteria culture obtained by culturing lactic acid bacteria in the above-mentioned medium for lactic acid bacteria.

Furthermore, the present invention is a composition characterized by containing lactic acid bacteria and the above-mentioned medium for lactic acid bacteria.

Furthermore, the present invention is a method for producing freeze-dried lactic acid bacteria cells, which comprises dispersing the lactic acid bacteria cells cultured using the above-mentioned medium for lactic acid bacteria in a dispersion medium and then freeze-drying them.

The medium for lactic acid bacteria of the present invention can promote the growth of all lactic acid bacteria and achieve a high number of bacteria after culture.

Moreover, since the medium for lactic acid bacteria of the present invention is composed of a food material, it can be used as it is in various foods and drinks after culturing.

The medium for lactic acid bacteria of the present invention (hereinafter referred to as "the medium of the present invention") contains soybean peptide and yeast extract.

The above-mentioned soybean peptide is not particularly limited as long as it is a peptide derived from soybean, but examples include peptides obtained by hydrolyzing soybean with enzymes, acids, etc. according to conventional methods. The molecular weight of this soybean peptide is also not particularly limited, but is, for example, about 10,000 or less. Note that the molecular weight of soybean peptides may be measured using gel filtration analysis. Further, one type or two or more types of these soybean peptides can be used. Among these soybean peptides, soybean peptone obtained by hydrolyzing soybean with an enzyme such as protease is preferred. As such soybean peptone, Bacterio-N SS-(B)K (registered trademark, Maruha Nichiro Co., Ltd.), soybean peptone (Organo Food Tech Co., Ltd.), etc. are commercially available.

The content of soybean peptide in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 20% by mass (hereinafter simply referred to as "%"), preferably 0.5 to 10%, particularly preferably 0.5%. ~5%.

The above-mentioned yeast extract is not particularly limited as long as it is an extract derived from yeast, and examples thereof include yeast autolytic fluid, yeast extract, and the like. The method for obtaining the yeast extract is not particularly limited, and examples include a method of extraction with a solvent such as hot water or alcohol. The yeast extract may be an autolyzed yeast liquid, a yeast extract itself, or a concentrated liquid or powder thereof. The type of yeast is not particularly limited, and examples thereof include beer yeast, baker's yeast, wine yeast, etc., with beer yeast being preferred. Moreover, these yeast extracts can be used alone or in combination of two or more. Among these yeast extracts, beer yeast extract is preferred. Examples of such brewer's yeast extract include Meest P1G (registered trademark, Asahi Beer Foods Co., Ltd.), Meest P2G (registered trademark, Asahi Beer Foods Co., Ltd.), Yeast Extract B2 (Oriental Yeast Industries Co., Ltd.), etc. It is commercially available.

The content of yeast extract in the medium of the present invention is not particularly limited, but is, for example, 0.1 to 20% as yeast extract powder, preferably 0.5 to 10%.

The medium of the present invention may further contain one or more selected from the group consisting of fermented barley extract, emulsifiers, and sugars, preferably all of them.

The above-mentioned fermented barley extract is not particularly limited as long as it is an extract derived from fermented barley, but examples include the residual liquid after distillation of alcoholic beverages obtained when barley is fermented to produce alcoholic beverages such as shochu. , specifically, barley shochu distillation residue and the like. Moreover, these fermented barley extracts can be used alone or in combination of two or more. Among these fermented barley extracts, barley shochu distillation residue is preferred. As such barley shochu distillation residue, Burlex (registered trademark, Sanwa Shurui Co., Ltd.), Burlex S (registered trademark, Sanwa Shurui Co., Ltd.), etc. are commercially available.

The content of fermented barley extract in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 50% as extract (liquid), preferably 1.0 to 25%.

The emulsifier is not particularly limited, and examples thereof include polyglycerin fatty acid esters such as decaglycerin monooleate and decaglycerin monostearate. Moreover, these emulsifiers can be used alone or in combination of two or more. Among these emulsifiers, hydrophilic polyglycerol fatty acid esters are preferred among polyglycerol fatty acid esters, and decaglycerin monooleate is particularly preferred. As this decaglycerin monooleate, Sunsoft Q-17S (registered trademark, Taiyo Kagaku Co., Ltd.), NIKKOL DECAGLYN 1-OV (Nikko Chemicals Co., Ltd.), and the like are commercially available.

The content of the emulsifier in the culture medium of the present invention is not particularly limited, but is, for example, 0.001 to 5%, preferably 0.01 to 0.5%.

The above-mentioned saccharide is not particularly limited as long as it can be assimilated by lactic acid bacteria, and examples thereof include lactose, glucose, fructose, sucrose, and the like. Moreover, these saccharides can be used alone or in combination of two or more. Among these sugars, glucose and/or lactose are preferred, and lactose is more preferred. As lactose, lactose GranuLac200 (Megglet Japan Co., Ltd.), Meggletose B200 (Megglet Japan Co., Ltd.), etc. are commercially available. As glucose, anhydrous crystalline glucose (Sanei Toka Co., Ltd.) and the like are commercially available.

The content of saccharides in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 20%, preferably 0.5 to 5%.

Furthermore, the culture medium of the present invention may contain minerals, components that do not inhibit the growth of lactic acid bacteria, and which improve operability (antifoaming agents, etc.).

Preferred embodiments of the culture medium of the present invention include the following.
Soybean peptide 0.1-20%, preferably 0.5-5%
Yeast extract 0.1-20%, preferably 0.5-10%
Fermented barley extract 0.1-50%, preferably 1.0-25%
Emulsifier 0.001-5%, preferably 0.01-0.5%
Sugars 0.1-20%, preferably 0.5-5%
Water remaining amount

The most preferred embodiment of the culture medium of the present invention has the following composition, and the culture medium with this composition is referred to as SMB medium.
Soy peptone 2%
Yeast extract 2%
Fermented barley extract 6%
Decaglycerin monooleate 0.1%
Lactose 2%
Water remaining amount

The culture medium of the present invention can be prepared by dissolving the above components in a medium base material such as water or milk, followed by pH adjustment, sterilization, etc. The pH may be adjusted by adding sodium hydroxide or the like, and the preferred pH is 6.0 to 7.2, and the more preferred pH is 6.2 to 7.0. Note that when saccharides are contained in the present invention, it is preferable to dissolve substances other than saccharides in the base material, and then dissolve the saccharides after sterilization.

Note that the culture medium of the present invention can also be made into a culture medium kit by encapsulating components other than the culture medium base material. Then, at the time of use, this culture medium kit may be appropriately dissolved in a base material.

The medium of the present invention described above can be used for culturing lactic acid bacteria. The culture conditions for lactic acid bacteria using the medium of the present invention are not particularly limited, and normal culture conditions for lactic acid bacteria can be applied. The types of lactic acid bacteria that can be cultured in the medium of the present invention are not particularly limited, and include lactic acid bacteria of the genus Lactobacillus, Enterococcus, Lactococcus, Streptococcus, and Bifidobacterium. Among these lactic acid bacteria, lactic acid bacteria of the genus Lactobacillus are preferred, and among the lactic acid bacteria of the genus Lactobacillus, Lactobacillus crispatus , Lactobacillus acidophilus , Lactobacillus casei , and Lactobacillus casei are preferred. Lactobacillus delbrueckii subsp . bulgaricus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp . lactis , Lactobacillus plantarum plantarum ), Lactobacillus gasseri, Lactobacillus acidipiscis, Lactobacillus brevis , Lactobacillus coryniformis , Lactobacillus delbrueckii subsp . Lactobacillus delbrueckii subsp. delbrueckii , Lactobacillus kefiri, Lactobacillus kefiranofaciens subsp. kefiranofaciens , Lactobacillus kefiranofaciens subsp. kefiranofaciens Lactobacillus kefranofaciens subsp. kefirgranum , Lactobacillus nodensis , Lactobacillus parabrevis, Lactobacillus paracasei , Lactobacillus parakefiri , Lactobacillus parakefiri・Lactobacillus pentosus , Lactobacillus perolens, Lactobacillus rhamnosus , Lactobacillus salivarius , Lactobacillus tucceti, Lactobacillus curvatus curvatus ), Lactobacillus johnsonii , Lactobacillus fermentum , and Lactobacillus mali . One or more of these lactic acid bacteria can be cultured. Among these, Lactobacillus crispertus, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus plantarum, and Lactobacillus gasseri are preferred; Lactobacillus crispertus is more preferred, and Lactobacillus crispertus YIT 12319 (FERM BP-11500, date of entrustment: April 28, 2011, Patent Organism Depositary, National Institute of Technology and Evaluation (Kisarazu City, Chiba Prefecture, Japan 292-0818) Kazusa Kamatari 2-5-8 Room 120)) is particularly preferred.

By culturing lactic acid bacteria in the medium of the present invention, a lactic acid bacteria culture can be obtained. The culture temperature may be set appropriately depending on the lactic acid bacteria, but 30 to 37°C is preferable. Since the culture medium of the present invention is composed of a food material, the lactic acid bacteria culture can be blended into conventional foods and drinks as is or after being concentrated, diluted, dried, etc. Furthermore, compositions containing lactic acid bacteria and the culture medium of the present invention can be similarly incorporated into conventional foods and drinks.

Foods and drinks to which the above composition can be blended are not particularly limited, but include processed meat foods such as ham and sausages, processed seafood foods such as kamaboko and chikuwa, bread, sweets, butter, and yogurt. Examples include fermented milk, soft drinks, dairy products, lactic acid bacteria drinks, and lactic acid bacteria drinks. In addition, the form of the food/beverage product includes commonly used forms of food/beverage products, such as solid forms such as powder and granules, paste forms, liquid forms, and the like. Furthermore, dried microbial cells may be processed into tablets, powders, chewables, hard capsules, soft capsules, pills, gums, and the like. Oral compositions include mouthwash, toothpaste, powdered toothpaste, water toothpaste, oral ointment, gel, tablet, granule, fine granule, gummy jelly, troche, tablet, capsule, candy, chewing gum, etc. It can also be used in pet food, etc.

The culture medium of the present invention can be used for producing freeze-dried lactic acid bacteria cells. The method for producing freeze-dried lactic acid bacteria cells is not particularly limited, and any known method for producing freeze-dried lactic acid bacteria cells can be used as appropriate, but preferably lactic acid bacteria cells cultured using the medium of the present invention are used. This is a method in which the product is dispersed in a dispersion medium and then freeze-dried.

The dispersion medium in which the lactic acid bacteria cells are dispersed may be appropriately selected depending on the type of lactic acid bacteria, but for example, it is preferable to use an aqueous solution containing a protective agent and an antioxidant. The method of dispersing lactic acid bacteria cells in a dispersion medium is not particularly limited, and may be performed by adding the lactic acid bacteria cells to the dispersion medium and stirring. Further, the amount of lactic acid bacteria cells to be dispersed in the dispersion medium is not particularly limited, but is, for example, about 0.01 to 20%. The lactic acid bacteria to be dispersed are basically live bacteria, but may also contain dead bacteria.

The protective agent used in the dispersion medium is not particularly limited, and includes, for example, glutamic acid, glutamic acid salts such as sodium glutamate and potassium glutamate, disaccharides such as trehalose, sucrose, lactose, and maltose, glycerol, maltodextrin, cyclodextrin, Examples include skimmed milk powder and potato starch. One or more of these protective agents can be used, but it is preferable to use trehalose, skim milk powder, and potato starch. The content of the protective agent in the dispersion medium is not particularly limited, but is preferably, for example, 1 to 40%, more preferably 3 to 40%, particularly preferably 3 to 30%.

The antioxidant used in the dispersion medium is not particularly limited, and examples include ascorbic acid, salts of ascorbic acid such as sodium ascorbate and calcium ascorbate, vitamin E, catechin, glutathione, and astaxanthin. These antioxidants can be used alone or in combination of two or more, but ascorbic acid is preferred. The content of the antioxidant in the dispersion medium is not particularly limited, but is preferably, for example, 0.01 to 10%, more preferably 0.05 to 5%.

The dispersion medium is preferably an aqueous solution containing one or more selected from the group consisting of skim milk powder, trehalose, ascorbic acid, and potato starch, more preferably an aqueous solution containing skim milk powder, trehalose, ascorbic acid, and potato starch. Particularly preferred are aqueous solutions containing 3-40%, 3-20%, 0.1-5% and 3-20% of milk powder, trehalose, ascorbic acid and potato starch, respectively. In addition, regarding a dispersion medium (potato starch, etc.) that does not affect the pH even when dissolved, the dispersion medium can be added after adjusting the pH of a suspension of lactic acid bacteria in the dispersion medium, which will be described later.

After the lactic acid bacteria cells are dispersed in the dispersion medium as described above, it is preferable to adjust the pH of the dispersion medium to an alkaline side, more preferably to adjust the pH to 7.5 or higher, and adjust the pH to 7.5 or higher. It is more preferable to adjust the pH to 5 to 10.5, and most preferably to adjust the pH to 8.0 to 10.0. The pH may be adjusted using an alkaline substance such as sodium hydroxide.

After adjusting the pH of the dispersion medium, it is freeze-dried. Freeze-drying conditions are not particularly limited, but for example, conditions such as freezing at -35°C to -45°C for 6 to 48 hours followed by drying at 12°C to 35°C for 40 to 90 hours. can be mentioned. An example of the freeze dryer is TF20-80TANNS (manufactured by Takara Seisakusho Co., Ltd.).

The freeze-dried lactic acid bacteria cells thus obtained can maintain a high bacterial count even after storage. Specifically, the number of viable bacteria after storing freeze-dried lactic acid bacteria cells at 37°C for 8 weeks is determined by dispersing the lactic acid bacteria cells in a dispersion medium, adjusting the pH of the dispersion medium to 7.0, and then freeze-drying the cells. The number of viable bacteria after storing the freeze-dried lactic acid bacteria cells at 37° C. for 8 weeks is 120% or more. Of course, in order to produce freeze-dried lactic acid bacteria cells obtained by adjusting the pH of the dispersion medium to 7.0, freezing of lactic acid bacteria obtained by adjusting the pH of the dispersion medium to 7.5 to 10.5 is required. The same conditions and strain as used for producing dried bacterial cells are used except for the pH of the dispersion medium. In addition, the survival rate (%) after storing lyophilized lactic acid bacteria cells at 37°C for 8 weeks was determined by dispersing the lactic acid bacteria cells in a dispersion medium, adjusting the pH of the dispersion medium to 7.0, and then lyophilizing the cells after lyophilization. It is more preferable that the survival rate of the freeze-dried lactic acid bacteria cells obtained by the above method is 110% or more when the survival rate after storage for 8 weeks at 37° C. is 100%. The survival rate is calculated as (number of viable bacteria after storage period/number of viable bacteria on day 0 of storage) x 100.

The freeze-dried lactic acid bacteria cells described above can be used in foods and drinks as they are or by mixing them with other food materials that are normally added to foods. For example, foods include processed meat foods such as ham and sausage, processed marine foods such as kamaboko and chikuwa, bread, sweets, butter, yogurt, and fermented milk. Beverages include soft drinks, dairy products lactic acid bacteria drinks, lactic acid bacteria drinks, and the like. In addition, the form of the food/beverage product includes commonly used forms of food/beverage products, such as solid forms such as powder and granules, paste forms, liquid forms, and the like. Furthermore, the dried microbial cells may be processed into tablets, powders, chewable preparations, hard capsules, soft capsules, pills, gums, and the like. Oral compositions include mouthwash, toothpaste, powdered toothpaste, water toothpaste, oral ointment, gel, tablet, granule, fine granule, gummy jelly, troche, tablet, capsule, candy, chewing gum, etc. It can also be used in pet food, etc.

EXAMPLES Hereinafter, the present invention will be explained in detail with reference to Examples, but the present invention is not limited to these Examples in any way.

Implementation example 1
Preparation of medium and culture of lactic acid bacteria:
The components listed in Table 1 below were mixed and dissolved in water, the pH was adjusted to 6.2 to 6.7 with sodium hydroxide, and the mixture was sterilized to prepare a medium. Lactobacillus crispertus YIT 12319 (FERM BP-11500, date of entrustment: April 28, 2011, Japan Patent Organism Depositary, National Institute of Technology and Evaluation (Kazusa, Kisarazu City, Chiba Prefecture, Japan 292-0818) Kamatari 2-5-8 Room 120) (hereinafter also referred to as "LcY") was inoculated at 0.25% and cultured at 37°C for 12 hours. Table 1 also shows the results of measuring the number of viable bacteria (cfu/ml) and pH after culturing. The number of viable bacteria was measured using a spiral plater (EDDY JET, IUL Instruments).

From this result, it was found that media 5 and 6 of the present invention containing both soybean peptide and yeast extract had a higher number of viable bacteria after culture than media 1 to 4.

Implementation example 2
Preparation of medium and culture of lactic acid bacteria:
A medium was prepared in the same manner as in Example 1 except for using the components listed in Table 2 below (this medium is referred to as "SMB medium"). This medium was inoculated with 0.25% of the lactic acid bacteria listed in Table 3, and cultured at 37°C for 12 or 24 hours. For comparison, lactic acid bacteria were cultured at 37° C. for 12 or 24 hours using MRS medium (Becton Dickinson & Company). The number of viable bacteria after culture was measured in the same manner as in Example 1, and the results are shown in Table 3.

From this result, it was found that the SMB medium yielded the same or higher number of viable bacteria than the MRS medium, which is a synthetic medium.

Implementation example 3
Preparation of medium and culture of lactic acid bacteria:
B. in the same SMB medium as in Example 2. breve YIT 12272 ^*11 or B. bifidum YIT 10347 ^*12 was inoculated at 1% and cultured under anaerobic conditions at 37°C for 12 or 24 hours. For comparison, lactic acid bacteria were cultured under anaerobic conditions at 37° C. for 12 or 24 hours using MRS medium (Becton Dickinson & Company). As a result of measuring the number of viable bacteria after culturing in the same manner as in Example 1, B. breve YIT 12272 and B. bifidum YIT 10347 was also confirmed to grow in SMB medium.
*11: FERM BP-11320, Entrusted date: February 16, 2010, Patent Organism Depositary Center, National Institute of Technology and Evaluation (Address: 2-5-8 120 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) Issue room )
*12: FERM BP-10613, date of entrustment: June 23, 2005, National Institute of Technology and Evaluation, Patent Organism Depositary (120 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) Issue room )

Implementation example 4
Production of freeze-dried lactic acid bacteria cells:
(1) Preparation of dispersion medium suspension The culture solution of LcY precultured in SMB medium was inoculated into 2 L of SMB medium at 0.1%, and then statically cultured at 37°C for 12 hours (until reaching the stationary phase). . Next, each culture solution was centrifuged (4°C, 4500 x g, 7 minutes), the bacterial cells were washed once with physiological saline, and the LcY bacterial cells were dissolved in a dispersion medium (20% skimmed milk powder, 10% trehalose, ascorbic acid). (pH 5.1). In the system in which the pH was not adjusted, 10% potato starch was further added and mixed to obtain a dispersion medium suspension. On the other hand, in the system for adjusting the pH of the dispersion medium, the pH was adjusted to pH 7.0 and 9.0 with 5N sodium hydroxide, and then 10% potato starch was added and mixed to obtain a dispersion medium suspension.

(2) Freeze drying and crushing The dispersion medium suspension obtained in (1) was frozen at -40°C for 2 days in a freeze dryer (TF20-80TANN: manufactured by Takara Seisakusho Co., Ltd.), and then at a shelf temperature of 20°C. Freeze-drying was performed for 2 days. This lyophilized product was pulverized to obtain lyophilized LcY cells.

(3) Storage test The LcY freeze-dried cells obtained in (2) were packaged into laminate bags with zippers (LamiZip, manufactured by Seisaku Nippon Sha Co., Ltd.), the air was thoroughly removed, and the zippers were closed. Thereafter, it was stored at 37°C for the period shown in Table 3, and the number of viable bacteria was measured before and after storage. The number of viable bacteria was measured using a spiral plater (EDDY JET, IUL Instruments). In addition, the survival rate was calculated using the following formula. Furthermore, when the number of viable bacteria without pH adjustment was taken as 100%, the ratio of the number of viable bacteria to the number of viable bacteria at pH 7.0 or 9.0 was calculated using the following formula. The results are also shown in Table 4.

From this result, it was found that increasing the pH of the dispersion medium before freeze-drying increases the number of viable bacteria and survival rate. In addition, the number of viable bacteria of the freeze-dried lactic acid bacteria cells after storage at 37° C. for 8 weeks was 20290% when the pH was adjusted to 7.0 and 100% when the pH was adjusted to 9.0.

Implementation example 5
Production of freeze-dried lactic acid bacteria cells:
Lyophilized LcY cells were obtained in the same manner as in Example 4, except that the static culture was performed at 30°C for 22 hours instead of at 37°C for 12 hours. The number of viable bacteria was measured for these in the same manner as in Example 4, and the survival rate was calculated. Furthermore, when the number of viable bacteria at pH 7.0 was taken as 100%, the ratio of the number of viable bacteria to the number of viable bacteria at pH 9.0 was calculated using the following formula. The results are shown in Table 5.

From this result, it was found that even if the culture conditions were changed, increasing the pH of the dispersion medium before freeze-drying increased the number of viable bacteria and survival rate.

Implementation example 6
Culture in tank:
160L or 900L of SMB medium was placed in a stainless steel tank with a capacity of 200L or 1000L, inoculated with 0.1% of precultured LcY, and cultured at 37°C under aerobic conditions for 12 hours. The same or higher number of viable bacteria was obtained compared to the medium.

The medium for lactic acid bacteria of the present invention can be used for culturing lactic acid bacteria.

Claims (6)

Lactic acid bacteria cells cultured in a lactic acid bacteria medium containing soybean peptide, yeast extract, fermented barley extract, emulsifier, and saccharide were mixed with 3-40% by mass of skim milk powder, 3-20% by mass of trehalose, and 0.1% ascorbic acid. The freeze-dried lactic acid bacteria cells are dispersed in a dispersion medium containing ~5% by mass and 3~20% by mass of potato starch, the pH of the dispersion medium is adjusted to 7.5 or higher, and then freeze-dried. Production method. The method for producing freeze-dried lactic acid bacteria cells according to claim 1, wherein the lactic acid bacteria belong to the genus Lactobacillus. The medium for lactic acid bacteria contains 0.1-20% by mass of soybean peptide, 0.1-20% by mass of yeast extract, 0.1-50% by mass of fermented barley extract, 0.001-5% by mass of emulsifier, and saccharides. The method for producing freeze-dried lactic acid bacteria cells according to claim 1 or 2, which contains 0.1 to 20% by mass. The method for producing freeze-dried lactic acid bacteria cells according to any one of claims 1 to 3, wherein the pH of the dispersion medium is adjusted to 8.0 to 10.0. The saccharide used in the medium for lactic acid bacteria is lactose, the fermented barley extract is barley shochu distillation residue, the soy peptide is soy peptone, the yeast extract is beer yeast extract, and the emulsifier is decaglycerin monooleate. A method for producing freeze-dried lactic acid bacteria cells according to any one of claims 1 to 4. The method for producing freeze-dried lactic acid bacteria cells according to any one of claims 1 to 5, wherein the lactic acid bacteria of the genus Lactobacillus are Lactobacillus crispertus and/or Lactobacillus casei.