JP2003250528A - Survival-improving agent of bacterium belonging to the genus bifidobacterium, proliferation-promoting agent, or method for producing fermentation product containing the bacterium - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a live Bifidobacterium bacterium-containing fermentation product having an excellent Bifidobacterium bacterium survival property, and to provide a food containing the same. <P>SOLUTION: A Bifidobacterium bacterium survival-improving agent contains as active ingredients the juice of one or more vegetables selected from pumpkin, cucumber, celery, eggplant, onion, garlic, avocado, grape, and persimmon, the ground products of one or more kinds of beans selected from red beans, white beans, large red beans, kidney beans, pea, violet Hana beans, Chana beans, black soybeans, blue soybeans, green gram, horse beans, Daifuku beans, Lens esculenta, and red lentil, the juice of violet sweet potato, the ground product of cocoa or powdered green tea, pyruvic acid, reduced glutathione, cysteine or β-carotene. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a survival improver for Bifidobacterium bacteria, a growth promoter, a method for producing a fermented product containing the same, a food or drink containing the same, and the production of the same. It relates to a method for improving the residual property.

[0002]

BACKGROUND ART Bifidobacterium and Lactobacillus bacteria have been shown to have a wide range of effects such as improvement of intestinal flora, improvement of fecal property, improvement of intestinal function, infection prevention, immunostimulation, cancer prevention. ing. It is considered that these microorganisms contribute to human health through improvement of the intestinal environment (edited by Tomohi Mitsuoka, Bifidobacterium genus, 1994; Yakult Central Research Institute, Lactobacillus).・ Casei Shirota strain, 1999).

Meanwhile, the Ordinance of the Ministry of Milk, etc. specifies that the number of viable lactic acid bacteria or yeast in fermented milk is 10 million cells / ml or more. Many foods containing live lactic acid bacteria, which are expected to have health effects, satisfy this viable cell count. Since Bifidobacterium is an obligate anaerobic bacterium, not only is it difficult to grow in the presence of oxygen, but it rapidly dies. Therefore, in order to provide Bifidobacterium bacteria to consumers in a lively state, the fermented product immediately after production is filled in a container made of an oxygen-impermeable packaging material,
Contact with oxygen must be completely cut off.

However, no oxygen-impermeable container is complete, the degree of freedom in molding is poor, and since composite materials are used, waste disposal is complicated, and such a container is expensive. Therefore, there are many restrictions on its use. Therefore, various studies have been conducted to maintain the viability of Bifidobacterium bacteria even under aerobic conditions. For example, N-acetylglucosamine, pantothenic acid or pantetheine, pantethein, peptides,
Survival agents such as lactulose have been reported. However, some of these are relatively expensive, and some of them have a great influence on the flavor when they are made into foods, and there is a demand for more superior materials, including in terms of price and survival effect. .

The survival of Bifidobacterium is considered to be influenced by the strain, growth phase, pH value of the product (food and drink), sugar added as a sweetener and dissolved oxygen. As an improvement method, the addition of yeast and lactic acid bacteria, the combined use of vitamin C, the material of the product container, etc. have been studied ("Bifidobacteria Science", Nos. 150-15).
Page 6. August 1, 1988. Yakult Honsha Co., Ltd.), a better survivability improving agent is demanded in order to cope with consumer preference that has been diversifying in recent years and to further improve survivability. On the other hand, in order for the Bifidobacterium bacterium to sufficiently exert the above physiological effects and the like, in products such as fermented milk, it is required to maintain a high viable cell count, but in general, to increase the viable cell count. Requires a long period of culture, which deteriorates workability. Therefore, spinach, broccoli and the like are used as growth promoters for Bifidobacterium (JP-A-11-266860) and potato juice (JP-A-6-21).
7733), the flavors of various materials may not be suitable for the flavor of fermented milk-containing foods containing bacteria of the genus Bifidobacterium, and depending on the type of material, survival after commercialization. Since some of them are not suitable for maintaining sex, more excellent growth promoters are required.

[0006]

SUMMARY OF THE INVENTION Therefore, the object of the present invention is to improve the preservation viability of live Bifidobacterium bacteria, which is expected to have health effects, and is inexpensive and easy to use in foods and the like. It is intended to obtain a material (survivability improving agent) and to provide a fermented product of a Bifidobacterium genus bacterium whose survival property is improved by the material, a food, a pharmaceutical product, and a cosmetic containing the same. Further, the present invention provides a growth promoter capable of increasing the number of viable bacteria of the genus Bifidobacterium inexpensively and easily, in particular, excellent growth in terms of flavor and survival improvement effect. It is intended to provide a promoter. A further object of the present invention is to provide a method for improving the survival of Bifidobacterium bacteria during storage by adding the obtained survival improving agent to foods and the like.

[0007]

Means for Solving the Problems As a result of intensive studies for solving the above problems, the present inventors have found that certain plant materials, compounds (biometabolites), etc. are fermented products containing Bifidobacterium bacteria. It was found that the viable cell count after storage can be stably maintained by adding the above-mentioned materials to the above, or by producing a fermented product of Bifidobacterium sp. It was also found that by adding a certain plant material during the fermentation of Bifidobacterium, it is possible to promote its growth, and at the same time, there is a material that also improves the survival property. Further, they have found that by using soy milk in combination with the above-mentioned material as a culture substrate for a fermentation product containing a Bifidobacterium bacterium, the survival properties of the bacteria are synergistically enhanced, and the present invention was completed. That is, the present invention specifically took the following means based on the above findings.

The survivability-improving agent for a Bifidobacterium bacterium according to claim 1 is one or more plant components selected from the following Group A or one biological metabolite selected from the following Group B: Alternatively, it is characterized in that two or more kinds are used as active ingredients. [Group A] Pumpkin, cucumber, celery, eggplant, onion,
Garlic, avocado, grape, oyster, adzuki bean, white adzuki bean, quince bean, kidney bean, pea, purple bean, green bean, black soybean, green soybean, mung bean, fava bean, daibu bean, lentil, red lentil bean, red bean sprouts, cocoa greens, cocoa. [Group B] Pyruvate, reduced glutathione, cysteine,
β-carotene.

The survivability-improving agent for a bacterium belonging to the genus Bifidobacterium according to claim 2 is, in the above-mentioned means, a squeezed product of one or more vegetables or fruits selected from the following A'group, and the following A ". Either one or two or more kinds of beans crushed from the group, one or more kinds of bio-metabolites selected from the following group B, squeezed product of Murasaki sweet potato, or cocoa or matcha ground matter [A'group] Pumpkin, cucumber, celery, eggplant, onion, garlic, avocado, grape, oyster [A'group] Azuki bean, white adzuki bean, quince bean, kidney bean, Peas, purple beans, green beans,
Black soybean, green soybean, mung bean, broad bean, daifu bean, lentil, red lentil bean. [Group B] Pyruvate, reduced glutathione, cysteine,
β-carotene

A viability-improving agent for a Bifidobacterium bacterium according to claim 3 is characterized in that, in any one of the above means, it further contains a soy milk component.

The method for producing a fermented product containing a Bifidobacterium bacterium according to claim 4, wherein the survival substrate for viability of a bacterium belonging to the genus Bifidobacterium is used as a survival substrate. Is added, and a Bifidobacterium bacterium is inoculated and cultured.

The method for producing a fermented product containing a Bifidobacterium bacterium according to claim 5 is characterized in that, in the above-mentioned means, the culture substrate on which the Bifidobacterium bacterium can grow contains soymilk or soybean protein. And The method for producing a fermented material containing a Bifidobacterium bacterium according to claim 6, wherein the bacterium belonging to the genus Bifidobacterium is Bifidobacterium breve or Bifidobacterium longum according to claim 4 or 5. Is characterized in that. A food or drink containing the fermented product containing a Bifidobacterium bacterium according to claim 7, comprising the fermented product containing a Bifidobacterium bacterium obtained by the method according to any one of claims 4 to 6. To do.

The method for improving the viability of a Bifidobacterium bacterium according to claim 8 is a method for improving the viability of a Bifidobacterium bacterium-containing fermented product before or after fermentation of the Bifidobacterium bacterium-containing fermented product according to any one of claims 1 to 3.
It is characterized by adding any of the survival improvers described.

The Bifidobacterium genus bacterial growth promoter according to claim 9 is one of plant components selected from the following group C:
It is characterized in that one kind or two or more kinds are used as active ingredients. [Group C] Pumpkin, cucumber, onion, garlic, tomato, coconut, peanut, adzuki bean, white adzuki bean, kidney bean, kidney bean, pea, purple soybean, ground bean, black soybean, blue soybean, mung bean, broad bean, daifu bean, lentil, red Lentil bean.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The agent for improving survival of a Bifidobacterium bacterium according to the present invention comprises one or more selected from plant components or biometabolites containing a carboxyl group. is there.

The plant components of the above-mentioned survival improver include vegetables such as pumpkin, cucumber, celery, eggplant, onion, garlic, avocado, azuki bean, white azuki bean, quinto bean, kidney bean, pea, purple bean, ground bean, Beans such as black soybean, green soybean, mung bean, broad bean, daifu kumame, lentil, red lentil bean, fruits such as grapes and oysters, purple sweet potato, cocoa, green tea, etc., and handling and survival improvement points etc. Therefore, cucumber, celery, avocado, legumes and purple sweet potato are preferable, and adzuki bean, white azuki bean, common bean, ground bean, mung bean, fava bean, daifu bean, red lentil bean, and purple sweet potato are preferable.

Further, beans, pumpkin, cucumber, onion and garlic are preferable because they have a survival promoting effect and a growth promoting effect at the same time.

The above-mentioned biometabolite is an organic substance which is present in an organism and contains one or more carboxyl groups which can be separated. Specifically, pyruvic acid, reduced glutathione, cysteine, β-carotene. , Or salts thereof and the like.

The use form of the above plant material is not particularly limited, and it is a squeezed product, an extract, a culture solution fermented with microorganisms, a dried product or a crushed product thereof, or a concentrate (liquid) thereof.
It can be used as a diluent (liquid) or the like. Among them, it is preferable to use a homogenized one in order to stabilize the survival property of the Bifidobacterium bacterium in the fermented product. Therefore, the edible portion or the non-edible portion is treated with water or solvent. It is desirable to prepare a processed product by processing by appropriately combining heat treatment, crushing treatment, adsorbent treatment, homogenization treatment and the like. Examples of the crushed product include a crushed product obtained by crushing a dried plant material with a mill or the like, a shredded crushed product obtained by crushing after shredding, and the like.

Specifically, beans, potatoes (purple potatoes), fruits, vegetables, cocoa, and matcha are shredded crushed products (or other crushed products) or suspended in water or hot water. A turbid suspension, a squeezed product obtained by pressing the shredded crushed material and separating and removing the fiber content by centrifugation, filtration, etc., a clarified juice from which a precipitate has been separated and removed, or a solubilized product obtained by heating an insoluble matter, etc. Can be used. Further, a treatment liquid obtained by preheating these homogenates, a treatment liquid saccharified with a microorganism or an enzyme, a dilute liquid diluted with water or hot water, or a degreased processed product can also be used. Beans, cocoa and matcha are preferably crushed products or suspensions thereof in water or hot water, and as potatoes, fruits and vegetables, squeezed products obtained by pressing shredded crushed products. It is preferable to use, and it is particularly preferable to use a fiber-free squeezed product in which the fiber content of the squeezed product is separated and removed by centrifugation, filtration or the like. In addition, beans can be used as a processed product such as arachi bean paste and boiled azuki beans, and cocoa and matcha may be commercially available products.

The forms of bio-metabolites include microorganisms such as bacteria, yeasts and filamentous fungi, extracts from biological materials such as plants and animals, microorganisms such as bacteria, yeasts and filamentous fungi, plants,
Products using enzymes such as animals, or preparations such as chemically synthesized products can be used. Further, these preparations can be used as purified products of biometabolites or as a mixture with other components.

Further, the growth promoter of the present invention includes pumpkin, cucumber, onion, garlic, tomato, coconut, peanut, adzuki bean, white azuki bean, quinto bean, kidney bean, pea, purple lentil,
Green beans, black soybeans, green soybeans, mung bean, fava beans, daifukumame, lentils, red lentil bean ingredients, among them, beans, pumpkin, cucumber, onion, garlic, as well as the growth-promoting effect and also the survival improvement effect. It is preferable because it has, and pumpkin and garlic are particularly preferable because they have a high growth promoting effect. The form of use of these growth promoters is the same as in the case of the above-mentioned survival improver, but it is preferable to prepare and use coconut milk as the coconut component and peanut butter as the peanut component according to a standard method.

The culture substrate for obtaining the fermented product containing viable bacteria of the genus Bifidobacterium is not particularly limited as long as it can grow the bacterium of the genus Bifidobacterium, plant material, animal material, microbial material, etc. However, those containing a soybean component or a milk component are preferable. Of these, soy milk, soy protein, cow milk, whole milk powder, skim milk powder, whey, or processed products thereof are more preferable, and soy milk and soy protein are particularly preferable.

Various materials can be used as additives for the culture substrate, and the following materials can be mentioned as examples. That is, glucose, sucrose, maltose, fructose, tagatose, lactose, glucose fructose syrup, trehalose, trehalulose, agarooligosaccharide, nigerooligosaccharide, galactooligosaccharide, fructooligosaccharide, xylooligosaccharide, raffinose, stachyose, lactulose, maltotriose, iso. Various sugars such as malto-oligosaccharide and cyclodextrin, various nutrients such as meat extract, yeast extract, fish meat extract, heart extract, liver extract, peptide, alginic acid, sodium alginate, fucoidan, sargassan, flucelan, funoran, porphyran, laminaran, pullulan , Tara gum, konjac mannan, inulin, β-glucan, chitin, chitosan, polydextrose, hyaluronic acid, chondroitin sulfate, manna , Galactan, fructan, xylan, arabinan, arabinogalactan, glucomannan, galactomannan, beet fiber, oat fiber, wheat fiber, soy fiber, rice fiber,
Barley fiber, xanthan gum, corn fiber,
Apple fiber, citrus fiber, silium fiber, pine fiber, prune fiber, banana fiber, acetic acid bacteria bacterial cellulose,
Lactobacillus cell wall, Bifidobacterium bacterium cell wall, yeast cell wall, natto fructan, collagen,
Various dietary fibers such as natto polyglutamic acid or various hydrolysates of these dietary fibers, wheat bran, barley bran,
Rice bran, oat bran, oat bran, rye bran, silium, rice bran, brown rice, chicory, soybean okara, apple pulp, resistant starch, barley malt, corn seed coat, lactic acid bacterium, bifidobacteria Fungus body, beer yeast fungus body, wine yeast fungus body, wine lees, sake lees, soy sauce lees, beer lees, rice malt, malt koji, soybean malt, red malt, yellow malt, natto mucilage, grape seed extract, royal jelly, Propolis, chlorella, spirulina, euglena, seaweed, kelp, kelp, hondawara, arame, kajime, asakusanori, aonori, hijiki, aosa, mozuku, etc. It can be cultured.

In addition, calcium, magnesium, zinc,
Iron, various minerals such as dolomite, or various salts of these minerals, citric acid, malic acid, tartaric acid, gluconic acid, succinic acid, fumaric acid, ascorbic acid, lactic acid,
Acetic acid, propionic acid, butyric acid, phosphoric acid, various acids such as amino acids or various salts of these acids, glutathione, phytin, phytic acid, lignin, saponin, ferulic acid, γ-aminobutyric acid, γ-oryzanol, chalcone, flavanone, flavone , Flavonols, isoflavones, anthocyans, catechins, proanthocyanidins, tea leaf polyphenols, curcumides, capsaicinoids, sesaminols, sesame lignans, theaflavins, β-diketones,
Carotenoids, allyl sulfur compounds, isothiocyanates, terpenes, chlorophylls, n-3 polyunsaturated fatty acids, n-6 polyunsaturated fatty acids, conjugated linoleic acids, phospholipids, plant sterols, eggs Various natural products such as protein, milk protein, rice protein, wheat protein, fish protein, vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, β-carotene, retinoic acid, folic acid, etc. Various vitamins, black cohosh, pumpkin seeds, pomegranate seeds, Hypericum perforatum, passionflower, valerian, Pueraria mirifica, rosemary, peppermint, parsley, marigold, lemon balm, mugwort, safflower, radish seeds, coffee tree, Ukogi, Yugao fruit, Kang Department of the pericarp, ginkgo leaf, Houttuynia, jujube, Kukoshi, licorice, Ganoderma lucidum, ginseng, various extracts, such as guarana, green tea, black tea, oolong tea, Tien-cha,
Gymnema tea, various plant extracts such as guava leaves, pepper,
Various spices such as salmon, cinnamon, turmeric, sage, thyme, basil, pepper and nutmeg may be added to the culture substrate for culturing.

Furthermore, rice, brown rice, barley, wheat, oats,
Rye, oats, germ components of seeds of various grains such as corn, corn, etc., ashitaba, kale, turmeric, potato, sweet potato, yam, eggplant, tomato, bitter melon, pepper, sesame, cabbage, broccoli, cauliflower, Lettuce, ginger, burdock, Japanese radish, horseradish, avocado, carrot, spinach, lily, lacquer, perilla, leek, leek, parsnips, bracken, bamboo shoots, shiitake, mushrooms, and other various vegetable ingredients, lemon, apple, strawberry, orange,
Guava, banana, blueberry, blackberry, cranberry, raspberry, cowberry, bayberry, feijoa, tamarillo, acerola, lime, shikuwasa, melon, peach, mango, yuzu, papaya, pineapple, pear, plum, grapefruit, karin, Fruit components such as apricots, mandarins, pomegranates, watermelons, prunes and kiwis, and various nut materials such as almonds, cashew nuts, macadamia nuts, chestnuts, ginseng and walnuts may be added to the culture substrate for culturing.

The viability-improving agent of the present invention may be added before, during or after the culturing of Bifidobacterium bacteria. To further enhance the effect, fermentation (culturing)
It is preferable to add it later. The growth promoter may be added before or during the culture of the Bifidobacterium bacterium, and is preferably added before the culture in order to enhance the effect.

The ratio of the additive added to the culture substrate may be any ratio, but the ratio of the solid content of the additive to the solid content of the substrate is preferably 99.999: 0.001 to 10:90. it can. For example, for sugars, 99: 1 to 90: 1
0 range, 99.99 to 0.01 to 99 to 1 for nutrients, 99 to 1 to 90 to 10 for dietary fiber or processed products or materials containing them, minerals and acids or their 99.99 to 0.01 to 99 to 1 for salts, 99.9 to 0.1 to 90 to 10 for natural products, 99.9 to 0.1 to 99 to 1 for vitamins, extracts or plant extracts 99.9 for food or spices
In the range of 0.1 to 99 to 1, 90 to 10 for grain ingredients, vegetable ingredients, fruit ingredients or nut ingredients
A range of 50 can be designated as a preferred range. Further, in order to adjust the optimum pH of the microorganism, an acid, a salt, a base or the like used in food may be added to the culture substrate for culturing.

The Bifidobacterium bacterium used in the production of a fermented product containing a viable bacterium of the Bifidobacterium bacterium may be of any kind as long as it is a microorganism belonging to the genus Bifidobacterium. The main member, Bifidobacterium breve
eve), Bifidobacte
rium longum), Bifidobacterium infantis, Bifidobacterium adolescent
is), Bifidobact
erium bifidum), Bifidobacterium catenulatum (Bifidobacterium catenulatum), Bifidobacterium pseudocatenulatum (Bifidobacterium)
pseudocatenulatum), Bifidobacterium angulatum (Bifidobacterium angulatum) and the like are preferable, and from the aspects of survival improvement effect and growth promotion effect, Bifidobacterium breve or Bifidobacterium
Longum is more preferable, and Bifidobacterium breve is particularly preferable.

Further, Bifidobacterium allicum, which is a Bifidobacterium bacterium having a source isolated from the human intestine, and Bifido, which is a Bifidobacterium bacterium used in foods. Bifidobacterium lacti
s), Bifidobacte animalis
rium animalis) etc. can also be used. For pets and livestock, Bifidobacterium pseudolongum, Bifidobacterium globosum,
Bifidobacterium sui
s), Bifidob thermophilus
Acterium thermophilus) etc. can also be used.

In the production of the fermented product containing the Bifidobacterium bacterium according to the present invention, a microorganism other than the Bifidobacterium bacterium can be used in combination. For example, Lactobacillus acidophilu
s), Lactobacillus gasser
i), Lactobacillus mali,
Lactobacillus plant
arum), Lactobacillus b
uchneri), Lactobacillus c
asei), Lactobacillus johnsonii (Lactobacillu
s johnsonii), Lactobaci galanum (Lactobaci)
llus gallinarum), Lactobacillus amylovorus, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus kefiroba, lactobacillus, lactobacillus Lactobacillus crispatu
s), Lactobacillus delbrueckii su
bsp. delbrueckii), Lactobacillus delbruecki subspecies bulgaricus
delbrueckii subsp. bulgaricus), Lactobacillus
Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus alimetarius
ntarius), Lactobacillus maltalomics (Lactoba
cillus maltaromicus) and other bacteria of the genus Lactobacillus, Streptococcus thermophilus (Streptococcus t
hermophilus) and other bacteria of the genus Streptococcus, Lactococcus lactis subspecies lactis (La
ctococcus lactis subsp.lactis), Lactococcus
Lactococcus cremoris (Lactococcu
lactis subsp. cremoris) and other bacteria belonging to the genus Lactococcus, Leuconostoc mesenteroides subs.
p. cremoris) etc., Bacillus subtilis (Bacillus subtilis), Bacillus coagulans (Bacillus coagulans) etc. Bacillus bacteria, Acetobacter aceti (Acetobacter aceti),
Acetobacter xylinum
Bacteria of the genus Acetobacter, Saccharomyces cerevisiae, Saccharomyces unisporus, Kluyveromyces marxianus (Kluyveromyces marx)
ianus), Torulaspora del Brocky (Torulaspora)
delbrueckii), Candida kefia
Yeasts belonging to the genera Saccharomyces, Torulaspora, Candida and the like such as yr) are mentioned as desirable microorganisms. Furthermore, filamentous fungi such as Aspergillus, Monascus, and Rhizopus can be used.

The culture conditions and method of the fermented product containing the Bifidobacterium bacterium are not particularly limited as long as the Bifidobacterium bacterium can be grown, and the culture substrate is inoculated with the bacterial solution of the cultivated microorganism. After that, fermentation may be performed by appropriately determining conditions such as temperature, time, stirring, and culture atmosphere suitable for the microorganism. For example, the culture temperature is preferably 30 to 37 ° C, and the culture time is preferably 15 to 48 hours.

The culture atmosphere is not particularly limited to the presence or absence of air, but it is preferable to use a sealed container that is not air-permeable during the culture, and a sealed culture medium containing an inert gas such as nitrogen is used. More preferably, the culture is performed using a container or a sealed container in which the gas phase on the culture substrate is replaced with an inert gas. Fermentation can be carried out under conditions suitable for culturing, for example, static culturing, stirring culturing, shaking culturing, aeration culturing and the like. The fermentation may be a mixed culture in which a plurality of strains of microorganisms are combined, or a continuous culture in which a plurality of strains are combined.

The timing of stopping the fermentation is not particularly limited, but it is usually continued until the viable number of Bifidobacterium bacteria is 1 × 10 ⁷ cells / ml or more, and then stopped by cooling or addition of an acid. The number of viable cells immediately after culturing is 1 x 1
Preferably due to the high effect in the case of 0 or ⁸ / ml, which is particularly noticeable in a case of adding various additives as food.

The thus obtained fermented product containing a viable bacterium of the bacterium belonging to the genus Bifidobacterium of the present invention may be used as it is as a food or an oral drug, but additives usually used may be added thereto. . Examples of additives used here include sugars, proteins, lipids, vitamins, extracts of various materials, fragrances, colorants, and thickeners.

For example, in addition to the above-mentioned sugars, various dietary fibers or their various hydrolysates, minerals, grains, honey, glucosamine, N-acetylglucosamine,
Various sugars such as, maple syrup, various sweeteners such as amazake, sorbitol, xylitol, erythritol, lactitol, various sugar alcohols such as palatinit, various high-sweetness sweeteners such as sucralose and aspartame, licorice, stevia, glycyrrhizic acid distribution Various natural sweeteners such as sugars, sucrose fatty acid esters, glycerin sugar fatty acid esters, various emulsifiers such as lecithin, agar, gelatin, carrageenan, guar gum, gum arabic, xanthan gum, pectin, locust bean gum, etc. ) Agents and the like may be added.

The fermented product containing viable bacteria of the genus Bifidobacterium of the present invention may be used as it is, or may be diluted with a liquid material and used. Further, it can be dried and processed into powder, granules, tablets, capsules and the like for use. In addition, further concentrated to concentrate,
It can also be provided after being processed into a high-viscosity liquid, gel, paste, sponge, solid or the like. Further, it can be frozen and processed into an ice shape, a sherbet shape or the like to be used and provided.

The fermented product containing live bacteria of the bacterium belonging to the genus Bifidobacterium of the present invention can be directly used as a food product, and can be added to all food products in an arbitrary range as the above-mentioned processed product. Preferred foods include milk and lactic acid beverages, milk such as yogurt, fermented milk, cheese, pudding, and ice cream, alcoholic beverages and soy milk, juice, tea, sweet drinks and other favorite beverages, raw sweets and semi-raw sweets, Japanese sweets, and Western sweets. , Uiro, rice cake, castella, jelly, dumpling,
Manju, sweet natto, yokan, cream puff, chocolate, gum, cake, candy and other confectionery, mayonnaise and other egg processed products, soup and other cooked processed products and nutritional drinks. When added to these foods,
The fermented product containing a viable bacterium of the bacterium belonging to the genus Bifidobacterium of the present invention can be contained in an amount of 1% to 99%.

The fermented product containing a live bacterium of the bacterium belonging to the genus Bifidobacterium of the present invention can also be used as medicines, cosmetics and the like. In that case, various excipients and lubricants,
It is possible to add materials commonly used in pharmaceuticals and cosmetics such as fragrances. The dose as a medicine varies depending on the administration method, age, weight and condition of the patient, but the daily intake of an adult patient is preferably 1 to 50 g as a solid content.

The fermented product containing a viable bacterium of the Bifidobacterium bacterium of the present invention is filled in a hermetically sealed container having low oxygen permeability composed of various materials suitable for stably maintaining living microorganisms. It can be provided, or can be provided by filling it in an oxygen-permeable sealed container, capsule, packaging bag, or the like. Further, it can be provided by filling the inside of the sealed container with an inert gas.

The storage of the fermented product containing the viable bacterium of the bacterium of the genus Bifidobacterium of the present invention is not particularly specified as long as it is a temperature of room temperature or lower, but is more preferably 10 ° C. or lower.

The fermented product containing a viable bacterium of the bacterium belonging to the genus Bifidobacterium of the present invention is not limited to humans, but can also be used for dogs, cats, cattle,
It can also be used for domestic animals such as horses, sheep, goats, kangaroos, pigs, chickens, ostriches, rabbits, guinea pigs, rats, mice, hamsters, lions, tigers, monkeys, chimpanzees, orangutans, gorillas, pet animals, and wild animals.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

[Example 1] Using soymilk (manufactured by Shikoku Kakoki, solid content 12%) as a culture substrate for Bifidobacterium, a plant preparation added at the time of culturing was found to have a survival property of Bifidobacterium. Was investigated.

(Investigation method) Tomatoes, onions, spinach, cabbage, broccoli, pumpkin, cucumber, ginger, celery, eggplant, peppers, garlic, avocado, bananas, oysters, cauliflower used as plants were used in the edible portion. Distilled water was added so that the solid content (4th edition Japanese Food Standard Ingredients Table (2nd edition), edited by the Science and Technology Agency Resource Research Committee) was about 10%, and the mixture was crushed with a mixer. Then, the mixture was filtered with gauze to obtain a filtrate as a plant preparation. The filtrates were used as additives during culture. As the coconut milk, a commercially available product was used as it was as an additive during culture. Peanut butter dispersed in 9 times the weight of water was used as an additive during culture. These culture additives were adjusted to pH 7 with 1N sodium hydroxide solution. A mixture was prepared as a culture sample by mixing soymilk as a culture substrate and the above-mentioned additives during culture in a weight ratio of 50:50. As a control sample, 50% diluted soy milk was used.

After adding 1% by weight of glucose to each culture sample, it was filled in a glass culture bottle and sterilized by steam at 121 ° C. for 15 minutes. After sterilization and rapid cooling in ice, the gas phase was filled with nitrogen gas and sealed with a butyl stopper. Of anaerobically prepared soy milk
Bifidobacterium breve YIT4065 (FERM BP-6223, hereinafter,
Abbreviated as 4065 shares. ) Was prepared, and 0.5% of the sterilized above-mentioned culture sample was inoculated under nitrogen gas, sealed with a butyl stopper, and cultured in an anaerobic state for 19 hours.

After culturing, the viable cell count and pH of the YIT4065 strain in these fermentation products were measured. For these fermented products, the viable cell counts were measured during anaerobic storage at 10 ℃ in a glass container and butyl stopper, or non-anaerobic storage at 10 ℃ in a glass container and silicon stopper. In addition, the measurement of the viable cell count is
The fermented product or the preserved fermented product was diluted appropriately, smeared on a MILS agar plate, anaerobically cultured at 37 ° C for 3 days, and then colonies were counted.

[0047]

[Table 1]

(Results) As shown in Table 1, among plant preparations mixed with soymilk, pumpkin, cucumber, celery, eggplant, onion, garlic, avocado, oyster had higher survival rate than the control 50% soymilk. Showed the rate. In a butyl stopper sealed state (anaerobic storage test) of each culture sample,
Pumpkin, cucumber, celery, onion, avocado,
For non-anaerobic preservation, cucumber, celery, eggplant, avocado,
The oysters were excellent. Cucumber, celery and avocado, which showed high survival properties under both conditions, were considered to be particularly excellent survival improvers. In addition, spinach, cabbage, broccoli, pumpkin, cucumber, onion, garlic, tomato, coconut milk, peanut butter and cauliflower had a growth promoting effect. Generally, when a growth promoter is added, the viability is deteriorated due to the decrease in the activity of the substrate due to the pH decrease of the substrate due to the acid production of the bacterium, but among these materials, pumpkin, cucumber, and onion. , Garlic was considered to be a particularly excellent growth promoter having a survival-improving effect as well as promoting growth.

[0049]

Example 2 The effect of the addition of plants and bio-metabolites as various materials to fermented soymilk of Bifidobacterium was investigated on the survival of Bifidobacterium.

(Investigation method) Plants among various materials added to fermented soybean milk include adzuki bean, white azuki bean, quinto bean, kidney bean, pea, purple soybean, green soybean, black soybean, blue soybean, mung bean, broad bean, daibukumame. , Red lentil bean, purple sweet potato, cocoa and matcha were used.

As the bio-metabolites, reduced glutathione, sodium pyruvate, cystine and β-carotene were used. For beans, dry matter is pulverized with a mill,
The preparation suspended in water and heated at 100 ° C. for 20 minutes was used as an additive to fermented soymilk. The purple sweet potato was squeezed from Ayamurasaki as an additive to the fermented soymilk. Cocoa and matcha were obtained by dissolving commercially available powder in water and used as additives to the fermented soymilk.
As the in-vivo metabolite, a reagent-level product was used as an additive to the fermented soymilk.

135 soy milk (made by Shikoku Kakoki, solid content 12%)
The plate was sterilized at 3.5 ° C for 3.5 seconds to obtain a medium. Separately, inoculate the 4065 strain inoculated with soymilk into the above medium under nitrogen gas.
% Inoculated and cultured at 34 ° C. for 20 hours. After the culturing, 0.2 volume of glucose-fructose syrup and 0.2 volume of agar sterilized by heating were added to 1 volume of the obtained fermented soy milk bacteria solution to prepare a fermented soy milk product. Various additives were added to the product so as to have the concentrations shown in Table 2 and mixed, then, dispensed into a paper container that was not oxygen impermeable and sealed with an aluminum cap. These samples were non-anaerobic preserved samples.

As for the additive-free sample in which the additive-containing solution was replaced with water, a sample was dispensed in a glass container and sealed with a butyl stopper to prepare an anaerobic storage sample.
After storage at 10 ° C for 21 days, the number of viable bacteria in the stored sample was measured by counting the colonies that appeared on the MILS agar plate.

[0054]

[Table 2]

(Results) As shown in Table 2, when unfermented fermented soymilk was stored for 21 days, the survival rate of the 4065 strain in the non-anaerobic container was about 0.04% to 1.3% as compared with the anaerobic container. Was reduced by. The survival rate of the 4065 strain at the time of non-anaerobic storage is
All beans, purple sweet potato, cocoa, green tea, reduced glutathione, sodium pyruvate, cysteine,
It was revealed that the addition was increased by adding β-carotene. Above all, the addition of beans, purple sweet potato, matcha, reduced glutathione, and sodium pyruvate was effective, and the effects of beans and purple sweet potato were particularly high. In addition, processed red bean such as sardine bean paste, boiled azuki bean, boiled azuki bean juice and the like also showed an activity of improving the survival property of the 4065 strain, and no disappearance of the activity due to processing was observed.

[0056]

Example 3 The effect of addition of adzuki bean powder to fermented milk of Bifidobacterium was investigated on the survival of Bifidobacterium.

(Investigation method) Azuki beans were made into powder by dry milling, then suspended in water and heated at 100 ° C. for 20 minutes. The preparation was used as an additive to fermented milk. Whole milk powder (made by Yotsuba Dairy Co., Ltd.) and yeast extract (Difco) were melted to 12% and 0.03%, respectively, and then plate sterilized at 135 ° C for 3.5 seconds to obtain a medium. Separately, 2% of the inoculum of strain 4065 prepared with a skim milk powder was inoculated into the above medium under nitrogen gas, and cultured at 37 ° C. for 20 hours.

After culturing, 0.2 volume of glucose-fructose syrup and 0.2 volume of agar solution were added to 1 volume of the obtained fermented milk bacterial solution to prepare a fermented milk product. After admixing 20 g of dry weight per 1 liter of the product as a dry weight, the mixture was dispensed into a paper container that was not oxygen impermeable and sealed with an aluminum cap. Further, the product without addition of Azuki was dispensed into a glass container and sealed with a butyl stopper to obtain an anaerobic storage sample. After storage at 10 ° C for 22 days, the viable cell count in the stored sample was measured by counting the colonies that appeared on the MILS agar plate.

[0059]

[Table 3]

(Results) As shown in Table 3, when the fermented milk without additives was stored for 22 days, the survival rate of the 4065 strain in the non-anaerobic container was reduced to about 0.7% as compared with the anaerobic container. did.
The survival rate of strain 4065 under non-anaerobic storage was increased by the addition of adzuki bean powder. The above results show that by adding adzuki bean to fermented milk, the survival of Bifidobacterium bacteria in storage under non-anaerobic conditions of the fermented bacteria-containing fermentation product of Bifidobacterium bacteria is improved. Is shown.

[0061]

[Example 4] The survival properties of the fermented soybean milk of Bifidobacterium produced using soybean milk to which the additive material was added were investigated.

(Investigation method) For Azuki, a dry matter was pulverized with a mill, suspended in water and heated at 100 ° C. for 20 minutes. In addition, the grape juice is 100 ° C of the juice of the berry A variety 20 ° C.
What was heated for a minute was used. A medium was prepared by adding 2% and 10% by weight of adzuki bean powder and grape juice to soymilk (manufactured by Shikoku Kakoki Co., Ltd., solid content: 12%) plate-sterilized at 135 ° C for 3.5 seconds. Separately, inoculum of 4065 strain inoculated with soy milk was inoculated into the above medium at 1% under nitrogen gas, and cultured at 34 ° C. for 20 hours. After culturing, heat-sterilized glucose fructose liquid sugar 0.2 volume and agar solution volume 0.2 volume were added to 1 volume of the fermented soybean milk bacterium solution obtained, and after mixing, dispensed into a paper container that is not oxygen impermeable. It was sealed with an aluminum cap. These samples were non-anaerobic preserved samples. As for the additive-free sample in which the additive-containing solution was replaced with water, a sample was dispensed in a glass container and sealed with a butyl stopper to prepare an anaerobic storage sample. After storage at 10 ° C for 21 days, the number of viable bacteria in the stored sample was measured by counting the colonies that appeared on the MILS agar plate.

[0063]

[Table 4]

(Results) As shown in Table 4, when the fermented milk without additives was stored for 21 days, the survival rate of the 4065 strain in the non-anaerobic container was up to about 0.2% of the survival rate in the anaerobic container. Decreased to. The survival rate of strain 4065 under non-anaerobic storage was increased by the addition of adzuki bean powder or grape juice.

The above results show that by adding adzuki bean or grape juice as soybean milk additive to the soybean milk, the fermentation of the fermented product containing live bacteria of the genus Bifidobacterium under non-anaerobic conditions was performed. It has been shown that the viability of Umbilus bacteria is improved.

[0066]

[Example 5] When fermented soymilk was produced with a bacterial species different from the above-mentioned bacterial species, the effect of addition of additional substances on the survival of Bifidobacterium was investigated.

(Investigation Method) Among the materials added to the fermented soymilk, Azuki was used as a representative material. For Azuki, a dry matter was pulverized with a mill, suspended in water and heated at 100 ° C. for 20 minutes to prepare an additive to the fermented soymilk. Soymilk (Shikoku Kakoki Co., Ltd., solid content 12%) was plate sterilized at 135 ° C for 3.5 seconds to obtain a medium. Separately, a 1% inoculum of Bifidobacterium longum ATCC 15707 strain prepared with soymilk was inoculated into the above medium under nitrogen gas, and cultured at 34 ° C. for 20 hours. After the culturing, 0.2 volume of glucose-fructose syrup and 0.2 volume of agar sterilized by heating were added to 1 volume of the obtained fermented soy milk bacteria solution to prepare a fermented soy milk product. After admixing 20 g of dry weight per 1 liter of the product as a dry weight, the mixture was dispensed into a paper container that was not oxygen impermeable and sealed with an aluminum cap. These samples were non-anaerobic preserved samples. As for the product without addition of azuki bean, it was dispensed into a glass container and sealed with a butyl stopper to prepare an anaerobic storage sample. 28 at 10 ° C
After storage for one day, the viable cell count in the stored sample was measured by counting the colonies that appeared on the MILS agar plate.

[0068]

[Table 5]

(Results) As shown in Table 5, when the additive-free fermented soymilk was stored for 28 days, the survival rate of the Bifidobacterium longum ATCC 15707 strain in the non-anaerobic container was the same as that in the anaerobic container. It was about 0.01% of the residual rate. It was revealed that the survival rate increased significantly when azuki bean was added and stored non-anaerobically. This result shows that when using adzuki bean as an additive, even fermented soy milk made with different bacterial species,
It has been shown that the viability of viable Bifidobacterium bacteria under non-anaerobic conditions is improved, and it is also shown that the type of Bifidobacterium bacteria does not matter.

[0070]

[Example 6] The effect of the fermented soybean milk of Bifidobacterium produced using soybean milk to which the additive material was added was investigated.

(Investigation Method) For Azuki, a dry matter was pulverized with a mill, suspended in water and heated at 100 ° C. for 20 minutes. A medium was prepared by adding 2% by weight of adzuki bean powder to soymilk (manufactured by Shikoku Kakoki Co., Ltd., solid content: 12%) that had been plate-sterilized at 135 ° C for 3.5 seconds. Another 4065 strain, Bi, made with soymilk
Fidobacterium catenulatum YIT4016 (hereinafter abbreviated as 4016 strain) inoculum was inoculated into the above medium at 0.5% and 2% respectively, non-anaerobically stoppered and cultured at 34 ° C. The effect of the additive on the growth property was to measure the pH over time, and the culture pH was 5.
Evaluation was made by culturing time until it reached 4.

(Results) pH 5.4 when cultured with soy milk only
The culture time of reaching 40% was 17 hours for the 4065 strain and 50 hours for the 4016 strain, but in the case of soy milk with adzuki bean added, it was shortened to 16 hours for the 4065 strain and 35 hours for the 4016 strain.

The results show that the use of adzuki bean as an additive during culture improves the growth of Bifidobacterium in soymilk under non-anaerobic conditions.

[0074]

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a Bifidobacterium bacterium, which is expected to have a health effect, in a state in which good survival is maintained even after storage, and on the other hand, Bifidobacterium It is possible to promote the growth of genus bacteria. In addition, as a result of the increased survival property, it is possible to use any container for storage and distribution of the fermented product containing Bifidobacterium bacteria regardless of oxygen permeability.

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Fumiho Ishikawa             1-1-19 Higashi-Shimbashi, Minato-ku, Tokyo             Yakult Head Office F-term (reference) 4B017 LC03 LK21 LP05                 4B018 LB08 MD85 MD91 ME11 MF13                 4B065 AA21X BB09 BB12 BB20                       BB26 CA42

Claims (9)

[Claims] 1. A bifidobacteria comprising one or more plant components selected from the following group A or one or more bio-metabolites selected from the following group B as an active ingredient. Survival-improving agent for Umbu bacteria. [Group A] Pumpkin, cucumber, celery, eggplant, onion,
Garlic, avocado, grape, oyster, adzuki bean, white adzuki bean, quince bean, kidney bean, pea, purple bean, green bean, black soybean, green soybean, mung bean, fava bean, daibu bean, lentil, red lentil bean, red bean sprouts, cocoa greens, cocoa. [Group B] Pyruvate, reduced glutathione, cysteine,
β-carotene. 2. One or more kinds of squeezed vegetables or fruits selected from the following A'group, one or more kinds of crushed beans selected from the following A "group, selected from the following B group: The bifidobacteria according to claim 1, characterized in that one or more kinds of biological metabolites to be used, a squeezed product of purple sweet potato, or a crushed product of cocoa or matcha is used as an active ingredient. Survival-improving agent for Umbu bacteria [A 'group] Pumpkin, cucumber, celery, eggplant, onion, garlic, avocado, grape, oyster [A "group] Azuki bean, white azuki bean, quinto bean, kidney bean, pea, purple saki Green beans, green beans,
Black soybean, green soybean, mung bean, broad bean, daifu bean, lentil, red lentil bean. [Group B] Pyruvate, reduced glutathione, cysteine,
β-carotene. 3. The viability-improving agent for Bifidobacterium bacteria according to claim 1 or 2, further comprising a soy milk component. 4. A method for adding a viability-improving agent according to claim 1 to a culture substrate on which a Bifidobacterium bacterium can grow, and inoculating and culturing the Bifidobacterium bacterium. A method for producing a fermented product containing a Bifidobacterium bacterium. 5. The method for producing a fermented product containing a Bifidobacterium bacterium according to claim 4, wherein the culture substrate on which the Bifidobacterium bacterium can grow is a soymilk- or soybean protein-containing culture substrate. Method. 6. The fermented product containing a Bifidobacterium bacterium according to claim 4 or 5, wherein the Bifidobacterium bacterium is Bifidobacterium breve or Bifidobacterium longum. Production method. 7. A food or drink comprising a fermented product containing a Bifidobacterium bacterium obtained by the method according to any one of claims 4 to 6. 8. A bifidobacteria characterized in that, before or after fermentation of a fermented product containing a Bifidobacterium bacterium, the viability-improving agent according to any one of claims 1 to 3 is added to a culture substrate. A method for improving the viability of Umbilus bacteria. 9. A growth-promoting agent for Bifidobacterium bacteria, which comprises one or more plant components selected from the following group C as an active ingredient. [Group C] Pumpkin, cucumber, onion, garlic, tomato, coconut, peanut, adzuki bean, white adzuki bean, kidney bean, kidney bean, pea, purple soybean, ground bean, black soybean, blue soybean, mung bean, broad bean, daifu bean, lentil, red Lentil bean.