WO2022185419A1 - 新規乳酸菌株、及び新規乳酸菌株を含有する組成物 - Google Patents

新規乳酸菌株、及び新規乳酸菌株を含有する組成物 Download PDF

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WO2022185419A1
WO2022185419A1 PCT/JP2021/008002 JP2021008002W WO2022185419A1 WO 2022185419 A1 WO2022185419 A1 WO 2022185419A1 JP 2021008002 W JP2021008002 W JP 2021008002W WO 2022185419 A1 WO2022185419 A1 WO 2022185419A1
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strain
cremoris
lactic acid
present
fermented
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French (fr)
Japanese (ja)
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真通 渡辺
英樹 小阪
知里 相磯
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Fujicco Co Ltd
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Fujicco Co Ltd
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Priority to JP2021559615A priority Critical patent/JP7035283B1/ja
Priority to CN202180095075.0A priority patent/CN117062905B/zh
Priority to PCT/JP2021/008002 priority patent/WO2022185419A1/ja
Priority to TW111106638A priority patent/TWI799153B/zh
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • 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; 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 novel lactic acid bacteria strains.
  • the present invention also relates to various uses of the lactic acid bacterium strain, such as compositions containing the lactic acid bacterium strain (inoculum, oral composition), and methods for producing fermented foods and drinks using the lactic acid bacterium strain.
  • lactic acid bacteria are widely used in the production of fermented milk and lactic acid beverages.
  • Lactococcus lactis subsp. cremoris a type of lactic acid bacterium, has long been used as a bacterium for producing fermented milk products such as yogurt and cheese. It is known that there are strains that produce it.
  • Yogurt obtained by fermenting Cremoris bacteria (hereinafter referred to as "Cremoris FC strain") isolated by the applicant as the main fermented lactic acid bacteria (trade name "Caspian Sea Yogurt", registered trademark of the applicant, registered No.
  • Cremoris FC strain-containing products such as inoculum for fermentation and supplements have been developed in anticipation of the above physiological effects.
  • lactic acid bacteria which are facultative anaerobes, are known to deteriorate in survival under aerobic or acidic (low pH) conditions, and the Cremoris FC strain is no exception. Therefore, in acidic foods such as yoghurt and dry powders such as lactic acid bacteria supplements, the development of Cremoris FC strain-containing products that suppress the decrease in the number of viable bacteria that occurs during long-term storage and maintain useful physiological effects is desired.
  • cremoris FC triggers IFN- ⁇ production from NK and T cells via IL-12 and IL-18”
  • the present invention provides a new Cremoris FC strain with improved acid tolerance (low pH tolerance) and improved survival under aerobic and dry conditions, in other words, fermentation
  • An object of the present invention is to provide a cremoris FC strain that has good survivability even in acidic foods and drinks such as milk and in a dry powder state.
  • the present invention provides an inoculum for fermentation, an oral composition such as supplements and fermented foods and drinks, and fermentation using the Cremoris FC strain, which has excellent acid resistance and aerobic conditions and excellent survival under dry conditions.
  • An object of the present invention is to provide a food and drink manufacturing method.
  • cremoris strain is the same mycological strain as the cremoris FC strain (Lactococcus lactis subsp. cremoris FC, FERM P-20185) owned by the applicant.
  • Cremoris FC strains Conventionally known Cremoris FC strains will be referred to as "Cremoris FC strains", "existing Cremoris FC strains” or “parent FC strains", and to distinguish the newly found Cremoris FC strains of the present invention from them, This Cremoris FC strain”, “Cremoris FC46 strain” or “FC46 strain”.
  • the present invention has the following embodiments.
  • Fermentation inoculum (II-1) Fermentation inoculum containing Lactococcus lactis subspecies cremoris FC46 strain (accession number: NITE BP-03310) or a subculture strain thereof.
  • II-2) The fermentation inoculum according to (II-1), which further contains Streptococcus thermophilus.
  • II-3 Lactococcus lactis subspecies cremoris FC46 strain (accession number: NITE BP-03310) or its subculture strain;
  • II-4) The fermentation inoculum according to any one of (II-1) to (II-3), which is in a dry state.
  • (III) Oral composition (III-1) An oral composition containing the lactic acid bacterium described in (I-1) or (I-2). (III-2) The oral composition according to (III-1), which is food, oral pharmaceuticals, or oral quasi-drugs. (III-3) The oral composition according to (III-1) or (III-2), which is a fermented food or drink.
  • the present Cremoris FC strain of the present invention has the same mycological properties and useful physiological actions as existing Cremoris FC strains, but is less viable under low pH conditions around pH 4 than existing Cremoris FC strains. It has high persistence and viscosity maintenance effect, and has excellent acid resistance.
  • the present Cremoris FC strain has higher survival under dry and aerobic conditions and is more resistant to dry and aerobic conditions than existing Cremoris FC strains. Therefore, the present Cremoris FC strain has excellent stability in oral compositions such as acidic foods and drinks such as yogurt and lactic acid bacteria beverages and dry supplements. Physiological action can be maintained and exerted for a long period of time.
  • FIG. 2 shows the results of measuring the number of viable cells (cfu/ml) of each Cremoris FC strain (FC46 strain, parent FC strain) in an acidic medium over time in Example 2.
  • cremoris FC strain This cremoris FC strain is a strain belonging to Lactococcus lactis subsp. cremoris having the following mycological properties.
  • Identification label Lactococcus lactis subsp. cremoris FC46, accession number: NITE BP-03310 Internationally deposited.
  • Carbon source utilization Assimilable to the following carbon sources: D-galactose, D-glucose, D-fructose, D-mannose, D-lactose, N-acetylglucosamine, aesculin.
  • cremoris FC strain has 100% homology with an existing registered cremoris FC strain (Lactococcus lactis subsp. cremoris FC, FERM P-20185). It was identified as a strain belonging to subsp. cremoris (Lactococcus lactis subsp. cremoris).
  • the nucleotide sequence of the 16S rRNA gene of this Cremoris FC strain is shown in SEQ ID NO: 1 of the sequence listing.
  • this Cremoris FC strain produces viscous polysaccharides like the existing Cremoris FC strains.
  • the present Cremoris FC strain like the existing Cremoris FC strain, has various useful physiological effects such as intestinal regulation, blood glucose level elevation suppression, immunostimulation, influenza virus infection protection, allergy suppression, and exercise condition improvement. presumed to have an effect.
  • the present Cremoris FC strain has higher survival and viscosity maintenance effects under low pH conditions around pH 4 compared to the existing Cremoris FC strain, and has acid resistance. It has the characteristic of being excellent in
  • the present Cremoris FC strain has higher survival under dry and aerobic conditions than the existing Cremoris FC strain, and has high survival under dry and aerobic conditions. It has the property of being highly resistant to Therefore, the present Cremoris FC strain has excellent stability in oral compositions such as acidic foods and drinks such as yogurt and lactic acid bacteria beverages and dry supplements. Physiological action can be maintained and exerted for a long period of time.
  • the Cremoris FC strain includes not only the Cremoris FC strain (deposited fungus) internationally deposited under the accession number: NITE BP-03310 and its original strain, but also its subculture strain.
  • the passage of this Cremoris FC strain can be carried out according to a standard method generally used for passage of lactic acid bacteria, and is not particularly limited. For example, a method of inoculating the present Cremoris FC strain into a test tube containing the TY-Glu medium (pH 7.0) and culturing at an optimum temperature of about 25 to 30° C. for about 12 to 48 hours can be mentioned.
  • Cremoris FC strains subcultured by culture can be added with glycerol to a final concentration of 10 to 30 w/v% and stored at -80°C until use.
  • a normal medium containing a carbon source, a nitrogen source, inorganic ions, and, if necessary, organic micronutrients can be used.
  • Any carbon source may be used as long as it contains an assimilable carbon source.
  • carbohydrates such as D-glucose, D-galactose, D-fructose, D-mannose, and D-lactose are preferably used.
  • As the nitrogen source assimilable nitrogen compounds or those containing them may be used, and examples thereof include ammonium sulfate, casamino acids, peptones, and the like.
  • Inorganic salts such as phosphoric acid, iron, potassium, magnesium, zinc, manganese, copper and calcium can also be used as appropriate. Furthermore, if necessary, amino acids, biotin, riboflavin, pyridoxine, nicotinic acid, pantothenic acid, folic acid, thiamine and other vitamins necessary for the growth of the fungus can be added to the medium and used.
  • the proliferation of the Cremoris FC strain can be enhanced by culturing at a temperature of 15°C or higher, preferably about 25-30°C, and at a pH of 5 or higher, preferably about pH 6-7.
  • the Cremoris FC strain can be cultured statically under aerobic conditions, but preferably under anaerobic conditions to enhance proliferation.
  • this Cremoris FC strain has high viability under dry and aerobic conditions. Alternatively, it may be dried together with other components such as medium components or excipients to prepare a dried product.
  • Fermentation inoculum also referred to as "lactic acid bacteria starter" for producing fermented foods and drinks by utilizing its fermentative ability.
  • Fermented foods include milk-based fermented foods (yogurt, lactic acid bacteria drinks, cheese, etc.), soymilk fermented foods and drinks mainly made from beans (yogurt, lactic acid bacteria drinks, etc.), grains (rice, Grain fermented food and drink (amazake, bread, etc.) and vegetable/fruit fermented food and drink (pickles, etc.) using wheat, etc. as the main raw material can be mentioned.
  • the fermentation inoculum of the present invention is preferably used for fermentation of raw materials mainly composed of milk or soy milk, and by fermentation of the raw materials, milk fermented foods and drinks such as yogurt and lactic acid bacteria drinks, or soy milk fermented foods and drinks are produced.
  • the raw material milk raw material, soymilk raw material
  • milk raw material, soymilk raw material mainly composed of milk or soymilk may contain protein derived from milk or soybeans (preferably soybeans), and may be milk or soymilk itself. Alternatively, it may be one from which lipids have been removed or reduced.
  • milk used in the present invention includes animal milk, such as cow's milk, goat's milk, sheep's milk, horse's milk, etc., and its form is not particularly limited.
  • raw milk whole milk, skim milk, whole milk powder, skim milk powder, etc.
  • similk used in the present invention includes, but is not limited to, soymilk obtained by squeezing soybeans after grinding soybeans and water. It is a liquefied emulsion (soybean emulsion), and the type of beans used as raw materials is not particularly limited, and any beans such as soybeans, adzuki beans, peas, broad beans, kidney beans, and mung beans may be used.
  • the inoculum for fermentation of the present invention is used so that the number of viable cells of the aforementioned Cremoris FC strain is about 1 ⁇ 10 5 to 1 ⁇ 10 9 (cfu/g) when producing fermented milk or soymilk fermented food. is preferably inoculated. Therefore, before use, the present Cremoris FC strain is cultured in advance using, for example, a milk raw material or a soy milk raw material, and the number of viable bacteria in the culture is about 1 ⁇ 10 7 to 1 ⁇ 10 9 (cfu/g). (inoculum composition).
  • FC strain is inoculated and cultured at the optimum temperature to obtain the above bacterial concentration.
  • the number of viable cells of this Cremoris FC strain can be measured by pour culture method (25°C) using TY-Glu medium (pH 7.0) and counting the number of grown colonies. Yes (colony count).
  • the number of bacteria and turbidity are correlated, if the correlation between the number of bacteria and turbidity (absorbance at a wavelength of 600 nm) is determined in advance, the number of bacteria can be counted by measuring the turbidity.
  • the inoculum for fermentation of the present invention may consist solely of the present Cremoris FC strain, or may be used in combination with other microorganisms having fermentation ability as long as the effects of the present invention are not hindered.
  • Other microorganisms include, but are not limited to, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. lactis biovar.
  • lactic acid bacteria belonging to the genus Streptococcus such as Streptococcus thermophilus
  • examples include lactic acid bacteria belonging to the genus Lactobacillus, such as Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus gasseri, and Lactobacillus acidophilus. These lactic acid bacteria can be used singly or in any combination of two.
  • the microorganism used in combination with the present Cremoris FC strain is preferably Streptococcus thermophilus (hereinafter referred to as "S. thermophilus"). As shown in Example 3, which will be described later, this Cremoris FC strain was able to grow S. cerevisiae under low pH conditions. By co-fermenting with Thermophilus bacteria, a higher number of viable bacteria can be maintained (improved survivability) and a decrease in viscosity after storage can be suppressed compared to single fermentation.
  • the present Cremoris FC strain: S. Thermophilus 1:0.01-100, more preferably 1:0.1-10 can be exemplified.
  • S. The number of viable B. thermophilus bacteria can be measured by colony counting by the pour culture method (37° C.) using BCP-added plate count agar agar medium (Nissui Pharmaceutical Co., Ltd.).
  • acetic acid bacteria can also be used as a microorganism used in combination with this Cremoris FC strain.
  • acetic acid bacteria include, but are not limited to, acetic acid bacteria belonging to the genus Acetobacter, Gluconobacter, Gluconacetobacter, and the like.
  • the fermentation seed fungus of the present invention is not particularly limited as long as it exhibits fermentation ability when used in the production of fermented foods and drinks. From the viewpoints of sanitation, quality retention, handleability, etc., it is preferably a dry solid form, more preferably a dry powder or a dry granule. It may be prepared as a dried product by drying treatment together with other components such as medium components or excipients.
  • Fermentation conditions (culture conditions) for the fermentation inoculum of the present invention include: culture temperature of 15° C. or higher, preferably about 25 to 30° C.; culture pH of 5 or higher, preferably about pH 6 to 7; culture time of 3 hours or more, preferably 8. Up to about 24 hours can be mentioned.
  • the oral composition of the present invention is characterized by containing the present Cremoris FC strain.
  • the oral composition is prepared in the form of a food or drink using an appropriate edible carrier (raw material for food or drink) in the same manner as ordinary food or drink.
  • the oral composition is prepared into a pharmaceutical form (supplement form, pharmaceutical form, quasi-drug form) using suitable pharmaceutical raw materials such as excipients or diluents that are pharmaceutically acceptable. be done.
  • This Cremoris FC strain like the existing Cremoris FC strain, has various physiological actions in the body (intestinal regulation action, blood sugar level elevation suppression action, immunostimulatory action, influenza virus infection prevention action, etc.) , allergy suppression and exercise condition improvement action).
  • the action is considered to be the action of the present Cremoris FC strain itself and/or the viscous polysaccharide produced by the present Cremoris FC strain. Therefore, the oral composition of the present invention preferably contains the present Cremoris FC strain in a viable state. However, it is desirable to contain as many Cremoris FC strains as viable cells, and this does not preclude containing the Cremoris FC strains in a dead state.
  • the oral composition of the present invention only needs to contain the present Cremoris FC strain, and as long as it contains the present Cremoris FC strain culture, crude or purified culture product, freeze-dried product or spray-dried product thereof may be
  • the above culture is not limited, but is cultured for about 12 to 48 hours at an optimum temperature of about 25 to 30° C. using a medium suitable for the present Cremoris FC strain, such as TY-Glu medium (pH 7.0).
  • TY-Glu medium pH 7.0
  • the cells of the present Cremoris FC strain can be obtained by centrifuging the culture after the above culture, for example, at 3,000 rpm for about 10 minutes to collect the cells. These can also be purified according to a conventional method.
  • the above-mentioned cells can be freeze-dried or spray-dried.
  • the dried cells thus obtained can also be used as an active ingredient of the oral composition of the present invention.
  • the present Cremoris FC strain, its culture, crude and refined culture products, and dried products thereof are collectively referred to as "this Cremoris FC strain and its culture”.
  • the oral composition of the present invention can contain an appropriate amount of nutritional components suitable for maintaining and growing the Cremoris FC strain.
  • the nutrient components include carbon sources such as glucose, galactose, fructose, lactose and mannose used in culture media for culturing microorganisms; nitrogen sources such as yeast extract and peptone; vitamins and minerals; Each component such as a class, a trace metal element, and other nutritional components can be mentioned.
  • the carrier used in the preparation of each of these forms may be an edible carrier or a carrier such as a pharmaceutically acceptable excipient or diluent. Details of preparation into a food and drink form and edible carriers that can be used at that time are described later in the section "Oral compositions in food and drink form”.
  • the amount of the Cremoris FC strain to be incorporated into the oral composition of the present invention is generally such that the viable cell count of the Cremoris FC strain is 1 ⁇ 10 7 to 1 ⁇ 10 11 (cfu) in the oral composition of the present invention. ) can be appropriately selected from the amounts that are before and after.
  • the amount of the present Cremoris FC strain to be blended can be appropriately changed according to the form of the oral composition of the present invention to be prepared, the desired effect, and the like, using the above amount as a guideline.
  • Oral composition in the form of food or drink may be ingested as food or drink.
  • preferred examples include fermented milk such as yogurt, fermented milk foods and drinks such as lactic acid beverages, and soy milk such as soy yogurt and fermented soy milk beverages.
  • fermented milk such as yogurt
  • fermented milk foods and drinks such as lactic acid beverages
  • soy milk such as soy yogurt and fermented soy milk beverages.
  • a fermented food and drink can be illustrated.
  • formulation forms such as solid formulations (tablets, pills, powders, granules, microcapsules, capsules, etc.) and liquid formulations (solutions, suspensions, syrups, emulsions, etc.) containing this Cremoris FC strain
  • confectionery such as gum, caramel, gummy, nougat, and chocolate containing the present Cremoris FC strain of the present invention
  • fermented food and drink such as fermented vegetable drinks and fermented fruit drinks
  • Examples include dairy products.
  • the terms "fermented milk” and “lactic acid beverage” are defined in the former Ministry of Health and Welfare "Ministerial Ordinance Concerning Milk and Dairy Products Ingredients” Article 2, 37 “fermented milk” and 38 "lactic acid beverage”. shall be That is, the term “fermented milk” refers to paste or liquid obtained by fermenting milk or dairy products with lactic acid bacteria. Therefore, the fermented milk includes the yogurt form as well as the drink form.
  • lactic acid bacteria beverage refers to a beverage prepared by diluting a paste or liquid obtained by fermenting milk or dairy products with lactic acid bacteria as the main raw material with water.
  • the method for producing a fermented food and drink of the present invention is characterized by containing the present Cremoris FC strain.
  • a fermented food or drink can be produced according to a standard method, and includes, for example, appropriate fermentation raw materials (e.g., milk, soy milk (soybean emulsion), grains, vegetables, and fruits) containing nutrients of the present Cremoris FC strain. It can be produced by inoculating the present Cremoris FC strain into a raw material), culturing it, and fermenting the raw material.
  • Fermentation using the present Cremoris FC strain is preferably carried out by preparing a starter in advance and inoculating it into the raw materials for fermentation to ferment it.
  • the starter include the above-described fermentation inoculum of the present invention.
  • Such a starter preferably contains about 1 ⁇ 10 7 to 1 ⁇ 10 9 (cfu/ml) viable cells of the present Cremoris FC strain.
  • the raw materials for fermentation include, if necessary, fermentation promoters for good growth of the present Cremoris FC strain, such as carbon sources such as glucose, galactose, fructose, lactose and mannose; nitrogen sources such as yeast extract and peptone; Source: vitamins, minerals, trace metal elements, other nutritional ingredients, etc. can be added.
  • fermentation promoters for good growth of the present Cremoris FC strain such as carbon sources such as glucose, galactose, fructose, lactose and mannose; nitrogen sources such as yeast extract and peptone; Source: vitamins, minerals, trace metal elements, other nutritional ingredients, etc. can be added.
  • the inoculum amount of the present Cremoris FC strain is, for example, when milk is used as a raw material for fermentation, the amount of the present Cremoris FC strain in the milk is 1 ⁇ 10 5 (cfu/ml) or more, preferably 1 ⁇ 10 7 to 1 ⁇ It is suitable to be selected from the amount contained around 10 9 (cfu/ml).
  • Culture conditions are generally selected from a fermentation temperature of 15° C. or higher, preferably about 25 to 30° C., and a fermentation time of about 3 to 24 hours.
  • the fermented product obtained as described above has a curd form (yogurt or pudding-like form), which can be ingested as it is as a solid food.
  • the curd-form fermented product can be made into a desired beverage form by further homogenizing it. This homogenization can be carried out using a common emulsifier (homogenizer). Such homogenization makes it possible to obtain a beverage with a smooth texture.
  • homogenization if necessary, it is diluted appropriately, organic acids are added for pH adjustment, sugars, fruit juices, thickeners, surfactants, flavorings, etc. Various additives that are commonly used can be added as appropriate.
  • the beverage thus obtained can be aseptically filled into suitable containers for the final product.
  • the intake (administration) amount is appropriately determined according to the age, sex, weight, degree of disease, etc. of the living body to be ingested, and is not particularly limited.
  • the concentration of the Cremoris FC strain should be selected from the range of about 1 ⁇ 10 7 to 1 ⁇ 10 11 (cfu/ml).
  • the composition can generally be prepared so that about 50-1,000 mL thereof is ingested per person per day.
  • oral compositions of the present invention in the form of food and drink include supplement forms, specifically solid preparations such as tablets, pills, powders, granules, microcapsules and capsules; foods and drinks having formulation forms such as liquid formulations such as formulations, syrups and emulsions.
  • the food and drink having a formulation form is preferably a dry solid formulation.
  • Food and drink in the form of supplements like pharmaceutical compositions, contain fillers, extenders, binders, wetting agents, disintegrants, surface active agents, and lubricants that are usually used in this field as formulation carriers. It can be manufactured using a diluent or excipient such as an agent.
  • the amount of the lactococcus of the present invention to be added to the oral composition in the form of a food or drink may be any amount as long as the action of the lactococcus is exhibited, and can be appropriately selected from a wide range as long as the amount is sufficient. Usually, it is preferable to contain about 1 ⁇ 10 7 to 1 ⁇ 10 11 (cfu) in one ingestion (administration) unit form.
  • the method of administration is not particularly limited, and is determined according to various forms, age, sex and other conditions of the subject, degree of disease, and the like.
  • the amount of intake of the above composition is appropriately selected depending on the usage, age, sex and other conditions of the subject. It is preferably dosed at a rate of about 10 11 (cfu).
  • the intake of the present Cremoris FC strain, which is an active ingredient, is preferably about 1 ⁇ 10 7 (cfu) or more per day, and can be ingested in several divided doses per day.
  • the oral composition of the present invention is preferably taken continuously in order to obtain its effect effectively. For example, it can be exemplified a form in which it is ingested continuously or intermittently for two weeks or longer.
  • the oral composition of the present invention includes foods and drinks that have specific functions and are eaten for the purpose of maintaining health.
  • the oral composition of the present invention is a food or drink having an intestinal regulation effect, a blood glucose level elevation suppressing effect, an immunostimulatory effect, an influenza virus infection protective effect, an allergy suppressing effect, or/and an exercise condition improving effect. is included.
  • these foods and drinks include foods and drinks that are manufactured and sold by labeling or claiming the above effects and uses, such as foods with health claims (foods with nutrient function claims, foods with function claims, foods for specified health uses), foods for special dietary uses (specified health foods), and similar health foods.
  • Example 1 Isolation and identification of novel lactic acid bacterium Cremoris FC strain Cremoris FC strain (FERM P-20185) was inoculated (1%) into a sterilized 8% skim milk powder, and at its optimum temperature (25 ° C.) The mixture was left for about 10 hours to activate the Cremoris FC strain (starter 1).
  • inoculate (1%) Streptococcus thermophilus (hereinafter referred to as "thermophilus”) into a sterilized 8% skim milk powder, and keep it at its optimum temperature (37 ° C.) for about 10 hours. Leave to activate the thermophilus (starter 2).
  • starters 1 and 2 prepared above were inoculated into separately prepared sterilized milk, and fermentation was performed at 30°C. Cooling (4° C.) was initiated when curd formation was observed.
  • TY-Gal medium 1% galactose, 0.5% Bacto (TM) Tryptone (Thermo Fisher Scientific), 0.5% Bacto ( TM) Yeast Extract (Thermo Fisher Scientific), 0.5% NaCl) (pH 7.0) was inoculated at 4% and cultured at 26°C for 72 hours.
  • the culture solution was streaked on a TY-Gal agar medium (pH 7.0) and cultured at 25° C. for 48 hours, and grown colonies were cultured in sterilized 8% skimmed milk powder and preserved.
  • strains with high low pH resistance were screened by the method described in Example 1, and their mycological properties were evaluated according to standard methods.
  • Table 1 shows the results.
  • API50CHL manufactured by bioMérieux Japan Co., Ltd.
  • the assimilation ability for 49 types of carbon sources was evaluated.
  • Table 2 shows the results.
  • Table 1 is as described in paragraph [0015] and Table 2 is as described in paragraph [0016].
  • the obtained strain had the same mycological properties and sugar assimilation as the Cremoris FC strain used as the parent strain (hereinafter also simply referred to as "parent FC strain”).
  • parent FC strain the strain was identified as a strain belonging to Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. abbreviated as “Cremoris FC46 strain” or simply "FC46 strain”).
  • Cremoris FC46 strain was transferred to the National Institute of Technology and Evaluation Patent Microorganisms Depositary Center, which has an address in Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, as of November 6, 2020. , Identification mark: Lactococcus lactis subsp. cremoris FC46, Accession number: NITE BP-03310 Internationally deposited (deposit receipt date: November 30, 2020).
  • FC46 strain obtained above was added to TY-Glu medium (1% glucose, 0.5% Bacto (TM) Tryptone, 0.5% Bacto (TM) Yeast Extract, 0.5% NaCl) (pH 7.0). ) at a ratio of 4% and cultured at 25° C. for 15 hours to prepare a preculture solution.
  • the resulting preculture was inoculated into 40 ml of TY-Glu medium adjusted to pH 4.0 with lactic acid to a final concentration of 1%. This was statically cultured at 26° C.
  • FC46 strain in the culture medium was determined by the pour culture method using a BCP-added plate count agar agar medium (Nissui Pharmaceutical Co., Ltd.). Measurements were taken over time, and the survival rate (%) after 27 hours was calculated.
  • the parent FC strain was used in place of the above FC46 strain and cultured in the same manner. A residual rate (%) was calculated.
  • FIG. 1 shows the results of measuring the number of viable cells (cfu/ml) of each Cremoris strain (FC46 strain, parent FC strain) over time.
  • the parental FC strain rapidly died under low pH conditions of pH 4.0, and after 27 hours of culture, the number of viable cells was 6.6 ⁇ 10 3 (cfu/ml) (survival rate 0.2%).
  • the FC46 strain has a high survival rate even under low pH conditions of pH 4.0, and even after culturing for 27 hours under this pH condition, the survival rate is 1.9 ⁇ 10 6 (cfu/ml) (survival rate 33.6). %) was maintained. From this, it was confirmed that the FC46 strain is a cremoris strain that is superior in acid resistance compared to the parent FC strain.
  • Example 3 Evaluation of stability in fermented milk Fermented milk was prepared using the FC46 strain, and stability in the fermented milk was evaluated. As a control, fermented milk was similarly prepared using the parent FC strain, and the stability in the fermented milk was evaluated.
  • Each starter prepared above was inoculated into sterilized milk and fermented at 30°C. When the formation of curd was observed, cooling (4° C.) was started to suppress fermentation to obtain fermented milk.
  • pH, acidity, and viscosity in the fermented milk were measured by the following methods.
  • Measurement of pH The pH in the fermented product was measured after adjusting the fermented product to 10° C. using a pH meter F-52 (manufactured by Horiba, Ltd.).
  • Table 3 shows the results on the 1st day after the start of storage
  • Table 4 shows the results on the 30th day after the start of storage.
  • the parental FC strain died rapidly during storage, leaving only a few viable cells per mL after 30 days (fermented milk 1).
  • strain FC46 maintained a high viable count of 1.1 ⁇ 10 4 (cfu/ml) (survival rate 0.0026%) even after 30 days (fermented milk 2).
  • both the parent FC strain and the FC46 strain maintain a higher viable cell count than single fermentation by co-fermenting with Thermophilus (fermented milks 3 and 4), especially in the FC46 strain, 5.8 ⁇ 10 6 (cfu/ml) (survival rate 2.8%), which is a very high number of viable bacteria (fermented milk 4).
  • the parent FC strain showed a decrease in viscosity after storage for 30 days, but the decrease in viscosity was suppressed in the FC46 strain compared to the parent FC strain, and the same was true when co-fermented with Thermophilus. A tendency was obtained.
  • the fermented milks 1 to 4 produced in this example had good appearance (color, presence or absence of syneresis), aroma and taste, and no difference was observed in these respects.
  • Example 4 Preparation of Lactic Acid Bacteria-Containing Composition and Evaluation of Stability
  • a lactic acid bacterium-containing composition was prepared using strain FC46, and storage stability in a dry state was evaluated.
  • a composition containing lactic acid bacteria was prepared using the parent FC strain instead of the FC46 strain, and the storage stability in a dry state was similarly evaluated.
  • FC46 strain or parent FC strain was inoculated at a rate of 4% in Difco M17 medium (BD) supplemented with glucose to a final concentration of 0.5%, and incubated at 25°C for 16 hours.
  • a pre-culture was prepared by culturing. The same medium was inoculated with 4% of the preculture solution prepared above and cultured at 25° C. for 16 hours to obtain 10 L of culture solution. After subjecting this to centrifugation (3,000 rpm x 10 minutes) and removing the resulting supernatant by decantation, the residue was washed twice with physiological saline to concentrate the FC46 strain or the parent FC strain, respectively.
  • a lactic acid bacteria solution was prepared.
  • Skimmed milk powder was added to each prepared lactic acid bacteria solution at a rate of 10%, and this was further freeze-dried using a freeze dryer (FD-81, Tokyo Rikakikai Co., Ltd.) according to a conventional method to obtain about 50 g. Freeze-dried lactic acid bacteria (FC46 strain, parent FC strain) were obtained.
  • FC46 strain 0.5 g of the obtained freeze-dried lactic acid bacteria (FC46 strain, parent FC strain) was placed in a sterilized bag made of laminated film with a zipper (Lamizip [registered trademark]: manufactured by Seisan Seisan Co., Ltd.). and stored under dark conditions at 37°C (forced deterioration test). From the start of the test (first day), the number of viable bacteria of the FC46 strain or the parent FC strain in the freeze-dried lactic acid bacteria was measured over time, and the survival rate (%) was calculated.
  • FC46 strain died rapidly when stored at a temperature of 37°C, and the number of viable cells after 3 weeks was 3.2 ⁇ 10 9 (cfu/g) (survival rate: about 1.6%). In contrast, the FC46 strain showed a very high viable count of 1.1 ⁇ 10 10 (cfu/g) (survival rate: about 14.7%). From this, it was confirmed that the FC46 strain is superior in storage stability to the parent FC strain even in a dried powder state under aerobic conditions.
  • SEQ ID NO: 1 shows the nucleotide sequence of the 16S rRNA gene of this Cremoris FC strain.

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Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LINH, N. T. H. ET AL.: "Screening of lactic acid bacteria isolated from fermented food as potential probiotics for aquacultured carp and amberjack", FISHERIES SCIENCE, vol. 84, 2018, pages 101 - 111, XP036413567, DOI: 10.1007/s12562-017-1150-9 *
MORIYA, NAOKO; KANEMATSU, NOBUE; TAJIMA, KIYOSHI; NARITA, TAKUMI; KIMOTO-NIRA, HIROMI: "Feeding milk fermented with a Lactococcus strain to weaning piglets ameliorates growth performance, but not immune function", FINAL REPORTS FOR RESEARCH GRANTS FOR MEAT AND MEAT PRODUCTS, vol. 35, 30 November 2016 (2016-11-30), Japan, pages 345 - 350, XP009540748 *
ZHU ZHENGMING; YANG PEISHAN; WU ZHIMENG; ZHANG JUAN; DU GUOCHENG: "Systemic understanding ofresponse to acid stress using transcriptomics approaches", JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, BASINGSTOKE, GB, vol. 46, no. 11, 14 August 2019 (2019-08-14), GB , pages 1621 - 1629, XP036925464, ISSN: 1367-5435, DOI: 10.1007/s10295-019-02226-x *

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