JP6921350B1 - New lactic acid bacteria belonging to Lactobacillus plantarum isolated from Yaezakura flowers and their utilization - Google Patents

Abstract

PROBLEM TO BE SOLVED: To achieve a high number of viable bacteria to obtain an effect as a probiotic when fruits and vegetables are used as a fermentation substrate in the above-mentioned lactic acid bacteria which have been used for producing a fermented product of lactic acid bacteria. There wasn't. According to the present invention, a lactic acid bacterium solution cultured until it reaches a steady state is inoculated with 0.4% volume of a plant-based food or drink, and the reached viable cell count is 109 CFU when cultured at 30 ° C. for 48 hours. Regarding lactic acid bacteria that exhibit the property of / ml or more.

Description

The present invention was obtained from various fruit juices, vegetable juices, vegetable mixed juices, crushed fruits, novel lactic acid bacteria belonging to Lactobacillus plantarum excellent in fermentation of crushed vegetables, foods and drinks containing the above lactic acid bacteria, and the above lactic acid bacteria. It relates to a fermented product, a food or drink containing the fermented product, and a method for producing a fermented product using the above-mentioned lactic acid bacterium.

There are more than 100 trillion bacteria in the human intestine, and it is said that there are more than 1000 types. The bacterial species that has the effect of maintaining and improving the intestinal environment is lactic acid bacteria called good bacteria. Lactic acid bacteria not only suppress the growth of Escherichia coli and pathogens by producing lactic acid in the intestine and improve the intestinal environment, but also have physiological effects such as improvement of bowel movements, infection protection by increasing immunity, and antiallergic action. show.

For this reason, the intestine is called the second brain these days, and it has been said that maintaining a good intestinal environment leads to maintaining the health of the entire body.

Ingestion of lactic acid bacteria is important in normal daily life, but it is especially important to ingest lactic acid bacteria daily to maintain a good intestinal environment and raise immunity in the present day of corona.

However, after being ingested into the body and functioning in the intestine, lactic acid bacteria are excreted from the body without staying in the body.

For these reasons, daily intake of lactic acid bacteria is necessary to maintain a good intestinal environment.

Conventionally, in order to ingest lactic acid bacteria on a daily basis, it has become common to ingest lactic acid bacteria beverages or fermented milk obtained by fermenting milk using animal lactic acid bacteria.

The lactic acid bacteria used in such lactic acid fermented products are animal lactic acid bacteria suitable for fermentation of dairy raw materials, specifically, Lactococcus lactis, Lactobacillus bulgaricus, Lactobacillus acidophilus, Streptococcus thermos. For example, Philis.

On the other hand, there are people who are allergic to dairy products and cause indigestion due to dairy products, and it is difficult for such people to effectively ingest lactic acid bacteria from foods, so it is effective to ferment non-dairy products. It is necessary to develop a new lactic acid bacterium strain that can ingest lactic acid bacteria.

Recently, attention has been paid to plant-derived lactic acid bacteria capable of fermenting plant components. It is said that plant-derived lactic acid bacteria not only ferment plant components but also survive and proliferate in acidic regions, so that they reach the intestines alive and have the effect of directly maintaining and improving the intestinal environment. So far, lactic acid bacteria capable of fermenting plant components using materials other than dairy raw materials have been isolated, and lactic acid bacteria belonging to Lactobacillus plantarum are known as plant-derived lactic acid bacteria capable of fermenting soymilk. For example, the Lactobacillus plantarum HOKKAIDO strain coagulates into a solid yogurt when soymilk is fermented as a fermentation substrate, and this yogurt stimulates immune function and reduces stress markers (Unlicensed Document 1). Recently, Streptococcus salivarius Sakura 2 that ferments and coagulates soymilk in a short time has also been reported (Patent Document 1).

However, since there are not a few people who are allergic to soymilk and soymilk fermented products, fruits and vegetables are used as fermentation substrates as the third fermentation raw material by plant-derived lactic acid bacteria, which have a low protein content and are less likely to cause allergies. The development of fermented products has been desired.

In particular, if you can ferment a wide variety of fruits and vegetables, you will have more options to take lactic acid bacteria fermented products of other fruits and vegetables even if you are not good at one particular fruit or vegetable or have allergies. Almost all people will be able to ingest some form of fermented plant-derived lactic acid bacteria.

Lactobacillus plantarum belongs to Lactobacillus plantarum as a lactic acid bacterium having an excellent ability to ferment fruits and vegetables. For example Lactobacillus plantarum ATCC14431 as fermentation substrate those concentrated carrot juice was diluted 6-fold, it is known to reach the 10 ⁷ CFU / ml or more viable count (Patent Document 2).

Further, as lactic acid bacteria suitable for fermentation in fruit juice, Lactobacillus plantarum YIT0132, (FERM BP-11349), YIT0148 (FERM BP-11350) are known.

These lactic acid bacteria, the number reached bacteria when the grape and orange 100% juice was fermented substrate, it is both known to be more than 10 ⁸ CFU / ml. (Patent Document 3).

Japanese Unexamined Patent Publication No. 2020-61977 Japanese Unexamined Patent Publication No. 2001-252012 International Publication No. 2011/115114

Nishimura, et al. J Tradit Compliment Med. 2015 Aug 21; 6 (3): 275-80.

However, with the above-mentioned lactic acid bacteria used in the production of fermented lactic acid bacteria, when fruits and vegetables are used as fermentation substrates, it is not possible to achieve a high number of viable bacteria that can be obtained as a probiotic effect. rice field.

In addition, these strains belonging to Lactobacillus plantarum did not always give high concentrations of viable bacteria to various fruits and vegetables.

Therefore, the present invention belongs to Lactobacillus plantarum and has a high number of viable bacteria even when plants such as various fruit juices, vegetable juices, mixed vegetable juices, crushed fruits, and crushed vegetables are used as fermentation substrates. An object of the present invention is to provide a novel lactic acid bacterium having an excellent fermenting ability, a food or drink containing the lactic acid bacterium, and the like.

As a result of diligent studies to solve the above problems, the present inventor crushes not only 100% fruit juice, 100% vegetable juice, and vegetable mixed juice, but also fruit crushing, which is a lactic acid bacterium belonging to Lactiplantibacillus plantarum isolated from Yaezakura flowers. We have completed the present invention by finding that the ^{number of viable bacteria reached is 10 9} CFU / ml or more, which is capable of exerting a function as a probiotic, even when a product or a crushed vegetable product is used as a fermentation substrate.

That is, the present invention is a lactic acid bacterium that satisfies the following properties.
The lactic acid bacteria solution cultured until a steady state is reached the number of viable bacteria when cultured for 48 hours at 30 ° C. those 0.4% volume inoculation against plant food is 10 9 ^CFU / ml or more.

The present invention is orange, 100% fruit juice pine, tomato, 100% carrot vegetable juice, Lactobacillus plantarum reach viable count when the vegetable mix juice fermentation substrate is both 10 ⁹ CFU / ml or more It is a lactic acid bacterium belonging to (Lactobacillus plantarum).

Further, in the present invention, Lactobacillus plantarum (Lactobacillus plantarum) in which the number of viable bacteria reached when crushed fruits of strawberry, pear, watermelon, melon, mango, papaya, and red dragon fruit are used as fermentation substrates is 10 ^{9 CFU / ml or more.} It is a lactic acid bacterium belonging to Lactobacillus plantarum).

Further, the present invention relates to tomato, carrot, cabbage, lettuce, onion, radish, celery, Chinese cabbage, radish, turnip, red turnip, pepper, yellow paprika, sprout cabbage, zucchini, garlic, burdock, lentil, Yamato mana (Yamato vegetables). , Sirona (Yamato vegetables), Uda gold burdock (Yamato vegetables), Shukutsubomi (Yamato vegetables), Akane Katahira (Yamato vegetables) vegetables crushed the fermentation substrate and the time of arrival number of living bacteria are both 10 ⁹ CFU / It is a lactic acid bacterium belonging to Lactobacillus plantarum, which is more than ml.

Further, the present invention is a fermented product obtained by fermentation of a food material with the above-mentioned lactic acid bacteria.

Furthermore, the present invention is a food or drink containing the fermented product.

Furthermore, the present invention is a food or drink containing the above-mentioned lactic acid bacteria.

Further, the present invention is a method for producing a fermented product, which comprises inoculating a food material with the above-mentioned lactic acid bacteria and fermenting the food material.

The lactic acid bacterium of the present invention can ferment various plants, and can prepare foods and drinks such as beverages used for probiotics based on various fermentation substrates.

Hereinafter, embodiments of the present invention will be described. The following embodiments are examples, and the scope of the present invention is not limited to those shown in the following embodiments. It should be noted that the description of the same content will be omitted in order to avoid repetitive complications.

Definitions For convenience, the specific terms used in this application are collected here. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as generally understood by the parties to the technical field to which the present invention belongs. Unless otherwise specified in the context, the singular forms "a", "an" and "the" include multiple references.

The numerical ranges and parameters shown in the present invention are approximate values, but the numerical values shown in the specific examples are described as accurately as possible. However, each number essentially contains certain errors that inevitably result from the standard deviation found in each test measurement. Also, as used herein, the term "about" generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term "about" means that it is within acceptable standard error when considered by those skilled in the art.

The "vegetable" of the present invention means an edible plant, and includes edible parts such as leaves, roots, stems, flowers, buds, and fruits of the plant.

The "crushed product" of the present invention means a plant such as fruits and vegetables which is crushed as it is or by adding water to a juicer or the like and in a state where a solid component and a liquid component are mixed.

The "fermented product" of the present invention means a product fermented using lactic acid bacteria, and includes a product fermented using a fermentation substrate such as a food material as a raw material.

In the present specification, 100% fruit juice and 100% vegetable juice refer to those that satisfy the standard of 100% juice in the fruit juice and vegetable juice beverage quality labeling standards in the JAS method.

The method for producing the fruit juice may be either "straight fruit juice" in which the juice is squeezed and then frozen and stored, or "concentrated reduced fruit juice" in which the juice is concentrated by heating and then stored frozen. Further, sugars, flavors, oxidative preservatives and the like may be added as long as they meet the standards of 100% juice in the fruit juice beverage quality labeling standards in the JAS method.

Vegetable mixed juice is "Fulfilling vegetables (manufactured by Itoen)" "One day's worth of vegetables (manufactured by Itoen)" "Kinari vegetables (manufactured by Yakult Headquarters)" "One vegetable per day (manufactured by Kagome)" " "Kagome Vegetable Life (manufactured by Kagome)" and "48 kinds of dark vegetables (manufactured by Kirin)" were used.

Embodiment for carrying out the invention The lactic acid bacterium according to the present embodiment satisfies the following properties.
The lactic acid bacteria solution cultured until a steady state is reached the number of viable bacteria when cultured for 48 hours at 30 ° C. those 0.4% volume inoculation against plant food is 10 9 ^CFU / ml or more.

The number of viable bacteria reached is 1.0 × 10 ⁹ CFU / ml or more, 1.5 × 10 ⁹ CFU / ml or more, 2.0 × 10 ⁹ CFU / ml or more, 2.5 × 10 ⁹ CFU / ml or more, 3.0 × 10 ⁹ CFU / ml or more. , 3.5 × 10 ⁹ CFU / ml or more, 4.0 × 10 ⁹ CFU / ml or more, 4.5 × 10 ⁹ CFU / ml or more, 5.0 × 10 ⁹ CFU / ml or more, 5.5 × 10 ⁹ CFU / ml or more, 6.0 × 10 ⁹ CFU / ml or more, 6.5 x 10 ⁹ CFU / ml or more, 7.0 x 10 ⁹ CFU / ml or more, 7.5 x 10 ⁹ CFU / ml or more, 8.0 x 10 ⁹ CFU / ml or more, 8.5 x 10 ⁹ CFU / ml or more, It may be 9.0 × 10 ⁹ CFU / ml or more or 9.5 × 10 ⁹ CFU / ml or more.

The plant-based food and drink may be at least one selected from the group consisting of fruit juice, vegetable juice, mixed vegetable juice, crushed fruit, and crushed vegetable. The juice may be 100% fruit juice or less than 100% fruit juice. The vegetable juice may be 100% vegetable juice or less than 100% vegetable juice.

The fruit juice is not particularly limited, but fruit juices of grapefruit, orange, pine, apple and the like are preferable, and fruit juices of orange and pine are particularly preferable.

The vegetable juice is not particularly limited, but vegetable juices such as tomatoes and carrots are preferable.

The fruit that is the raw material of the crushed fruit is not particularly limited, but fruits such as apple, strawberry, pear, watermelon, melon, kiwi, mango, papaya, and red dragon fruit are preferable, and strawberry, pear, watermelon, melon, and mango are particularly preferable. , Papaya, red dragon fruit fruits are preferred.

Vegetables used as raw materials for crushed vegetables are not particularly limited, but tomatoes, carrots, cabbage, lettuce, cucumbers, onions, sardines, celery, Chinese cabbage, daikon, cubs, red cubs, peppers, yellow paprika, broccoli, asparagus, and Mizuna. , Shungiku, spinach, komatsuna, bok choy, nanohana, parsley, seri, bean seedling, sprout cabbage, bitter gourd, zucchini, ginger, garlic, burdock, renkon, creson, mana Yamato (Yamato vegetables), white vegetables (Yamato vegetables), Udakin Vegetables such as burdock (Yamato vegetables), buds (Yamato vegetables), and Akane Katahira (Yamato vegetables) are preferable, especially tomatoes, carrots, cabbage, lettuce, onions, Naganegi, celery, Chinese cabbage, daikon, cubs, red cubs, and peppers. , Yellow paprika, sprout cabbage, zucchini, garlic, burdock, renkon, mana Yamato (Yamato vegetables), Chinese cabbage (Yamato vegetables), Uda gold gobo (Yamato vegetables), buds (Yamato vegetables), Akane Katahira (Yamato vegetables) Vegetables are preferred.

The lactic acid bacterium solution may be obtained by culturing in Lactobacilli MRS Broth medium. The culture conditions are, for example, 30 ° C. for 16 hours.

In certain embodiments, the lactic acid bacterium exhibits at least one of the following properties (a), (b) and (c).
(A) Lactobacillus cultivated in Lactobacilli MRS Broth medium at 30 ° C. for 16 hours at 30 ° C. was added to 100% fruit juice of orange and pine, 100% vegetable juice of tomato and carrot, and 0. 4% volume inoculated reach viable count when cultured for 48 hours at 30 ° C. what was found at both 10 9 ^CFU / ml or more.
(b) Lactobacillus solution cultivated in Lactobacilli MRS Broth medium at 30 ° C. for 16 hours was applied to crushed fruits of strawberry, pear, watermelon, melon, mango, papaya, and red dragon fruit by 0.4 each. % volume reached viable count when cultured for 48 hours at 30 ° C. those inoculated is both 10 ⁹ CFU / ml or more.
(c) Tomato, carrot, cabbage, lettuce, onion, onion, celery, Chinese cabbage, radish, turnip, red turnip, pepper, yellow Paprika, sprout cabbage, zucchini, carrot, burdock, lotus root, mana Yamato (Yamato vegetables), Chinese cabbage (Yamato vegetables), Uda Kingobo (Yamato vegetables), turnips (Shukuri) (Yamato vegetables), Akane Katahira (Yamato vegetables) arrival number of viable bacteria are both 10 ⁹ CFU / ml or more when cultured for 48 hours at 30 ° C. those 0.4% volume inoculated respectively vegetable crushed.

The crushed fruit was adjusted as follows.
200 g of apple and 50 ml of water for crushed apple, 100 g of strawberry for crushed strawberry, 100 g of pear for crushed pear, 100 g of watermelon for crushed watermelon, 100 g of melon for crushed melon, 200 g of kiwi for crushed kiwi, For crushed mango, 200 g of mango and 100 ml of water, for crushed papaya, 100 ml of crushed papaya and 100 ml of water, and for crushed red dragon fruit, 150 g of red dragon fruit treated with a juicer at 85 ° C for 30 minutes. It was sterilized and adjusted.

The crushed vegetables were adjusted as follows.
Crushed tomato is 200 g of tomato, crushed carrot is 100 g of carrot and 100 ml of water, crushed turnip is 200 g of turnip and 100 ml of water, crushed lettuce is 200 g of lettuce, crushed cucumber is 200 g of cucumber, crushed onion. 150 g of onion, 100 ml of water for 200 g of Japanese mustard spinach, 50 ml of water for 100 g of celery, 150 ml of water for 150 g of bok choy, 100 ml of water for 200 g of Japanese radish, turnip crushed 100 g of turnip and 50 ml of water, crushed red turnip is 100 g of red turnip and 100 ml of water, crushed pepper is 150 g of pepper, crushed yellow paprika is 200 g of yellow paprika, and crushed broccoli is 100 g of broccoli and 100 ml of water. , Asparagus crushed product is 200 g of asparagus and 100 ml of water, Mizuna crushed product is 150 g of Mizuna and 150 ml of water, Shungiku crushed product is 150 g of Shungiku and 150 ml of water 200g, bok choy crushed product is 100g of bok choy and 100ml of water, nanohana crushed product is 100g of nanohana and 150ml of water, crushed parsley is 100g of parsley and 100ml of water 60 g of bean seedlings and 60 ml of water, crushed turnips in 100 g of turnips, 150 ml of water, crushed bitter gourd in 200 g of bitter gourd in 50 ml of water, crushed zucchini in 100 g of zucchini in 50 ml of water 50 ml of water, 100 ml of water for crushed garlic, 100 ml of water for 100 g of turnip, 100 ml of water for 100 g of turnip, 100 ml of water for 100 g of radish, 100 ml of water for 100 g of creson, Mana Yamato 70 g of Japanese mustard spinach and 70 ml of water, crushed Japanese mustard spinach 100 g of Japanese mustard spinach and 100 ml of water, crushed Japanese mustard spinach 100 g of Japanese mustard spinach and 100 ml of water, and crushed turnip crushed Japanese mustard spinach 100 g of water 50 ml of crushed Katahira Akane was prepared by treating 150 g of Katahira Akane with 150 ml of water with a juicer and treating at 85 ° C. for 30 minutes to sterilize.

In addition, in the present specification, the reached viable cell count is defined as the number of viable cells reached after culturing the inoculum at Lactobacillus MRS Broth (manufactured by Difco) medium autoclaved at 121 ° C. for 15 minutes at 30 to 37 ° C. until it becomes a steady state for 16 hours. , 0.4% volume of pre-cultivated solution is inoculated into a fermentation substrate such as 100% fruit juice, and the viable cell count after culturing at 30 ° C. or 37 ° C. for 48 hours is shown. The viable cell count is measured using an MRS agar plate. In addition, CFU / ml, which is a unit representing the number of viable bacteria reached, indicates the colony forming ability when 1 ml is inoculated on the culture plate.

In certain embodiments, the lactic acid bacterium belongs to the genus Lactobacillus. In another embodiment, the lactic acid bacterium belongs to Lactobacillus plantarum. In another embodiment, the lactic acid bacterium belongs to Lactobacillus plantarum Sakura 68.

In yet another embodiment, the lactic acid bacterium is a lactic acid bacterium deposited under accession number: NITE P-03374.

In certain embodiments, the lactic acid bacterium also includes a mutant strain capable of growing under the above conditions.

In order to obtain the physiological effect of the lactic acid bacterium of the present invention, for example, it can be mixed with a solid or liquid pharmaceutical non-toxic carrier and used in the form of a conventional pharmaceutical preparation. Examples of such a preparation include solid preparations such as tablets, granules, scattered and capsules, liquid preparations such as solutions, suspensions and emulsions, and freeze-dried preparations. These formulations can be adjusted by conventional means in the formulation. Examples of the above-mentioned pharmaceutical non-toxic carrier include fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, glucose, lactose, sucrose, starch, mannitol, dextrin, amino acid, gelatin and albumin. , Water, physiological starch and the like. Further, if necessary, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, tonicity agents, and excipients can be appropriately added.

Further, the lactic acid bacterium of the present invention can be contained not only in the above-mentioned preparation but also in any form of food or drink such as solid or liquid. When it is contained in foods and drinks, it may be contained as it is or together with various nutritional components. Specifically, when the lactic acid bacterium of the present invention is contained in a food or drink, an additive that can be used as a food or drink is appropriately used, and a form suitable for edible use is used by conventional means, that is, granular, granular, tablet, capsule. , It may be molded into a paste or the like. Types of food and drink include, for example, processed meat foods such as ham and sausage, processed marine foods such as kamaboko and chikuwa, foods such as bread, confectionery, butter, and milk powder, water, fruit juice, milk, soft drinks, and tea beverages. Beverages such as. The food and drink also includes animal feed.

In the present embodiment, the composition obtained by fermentation with the above-mentioned lactic acid bacteria is also provided. In the present embodiment, a composition containing the above-mentioned lactic acid bacteria and a food material is also provided. The food material may be a material derived from a plant. The plant-derived material may be plant leaves, roots, stems, flowers, buds, fruits, extracts thereof, crushed products thereof, and / or combinations thereof. The plant-derived material may be fruits, vegetables, extracts thereof, crushed products thereof, and / or a combination thereof.

In the present embodiment, foods and drinks containing the lactic acid bacteria or the composition are also provided. In the present embodiment, there is also provided a method for producing a composition, which comprises inoculating a food material with the above-mentioned lactic acid bacteria and fermenting the food material.

A fermented product obtained by fermentation of a food material with the lactic acid bacterium of the present invention, a food or drink containing the fermented product, and a food or drink containing the lactic acid bacterium of the present invention The fermented product of the present embodiment is obtained by fermentation of the food material with the lactic acid bacterium of the present invention. In addition, the food and drink of this embodiment contains a fermented product thereof. In the present embodiment, the food material is not particularly limited as long as it is used for food, and leaves, roots, stems, flowers, buds, fruits of plants, extracts thereof (for example, fruit juice), and crushed products thereof. In particular, those that can be fermented by the lactic acid bacterium of the present invention are preferable, and examples thereof include fruits, vegetables and soy milk. When a fermented fruit juice beverage is produced using the lactic acid bacterium of the present invention, the lactic acid bacterium of the present invention is first inoculated and cultured alone or simultaneously with other microorganisms in the fruit juice sterilized under the conditions suitable for the fruit juice to be used, and this is homogenized. Fermented fruit juice can be obtained by doing so. In the case of fruits, vegetables or soy milk, fermented vegetable juice, crushed fermented fruit, crushed fermented vegetables and fermented soy milk can be obtained by the same method as for fruit juice.

These fermented products may be mixed with unfermented fruit juice, unfermented vegetable juice, unfermented fruit crushed product, unfermented vegetable crushed product, alcohol, etc., and various nutrients, vitamins, flavors, etc. may be added to make the final product. It is possible. These are preferable because they are contained in a viable state of the lactic acid bacterium of the present invention.

The fruit juice used as a raw material for the fermented fruit juice beverage is not particularly limited, but fruit juices such as grapefruit, orange, pineapple, and apple are preferable, and orange and pineapple juices are particularly preferable.

The vegetable juice used as a raw material for the fermented vegetable juice is not particularly limited, but vegetable juices such as tomatoes and carrots are preferable.

The fruit that is the raw material of the fermented fruit crushed product is not particularly limited, but fruits such as apple, strawberry, pear, watermelon, melon, kiwi, mango, papaya, and red dragon fruit are preferable, and strawberry, pear, watermelon, and melon are particularly preferable. , Mango, papaya, red dragon fruit are preferred.

The vegetables used as raw materials for the above-mentioned crushed fermented vegetables are not particularly limited, but tomatoes, carrots, cabbage, lettuce, cucumbers, onions, onions, celery, Chinese cabbage, daikon, cubs, red cubs, peppers, yellow paprika, broccoli, and asparagus. , Mizuna, Shungiku, Chinese cabbage, Komatsuna, Chinese cabbage, Chinese cabbage, parsley, seri, bean seedlings, sprout cabbage, bitter gourd, zucchini, ginger, carrot, gobo, renkon, Chinese cabbage, mana Yamato (Yamato vegetables), Chinese cabbage (Yamato vegetables), Vegetables such as Uda Kingobo (Yamato vegetables), buds (Yamato vegetables), and Akane Katahira (Yamato vegetables) are preferable, especially tomatoes, carrots, cabbage, lettuce, onions, Naganegi, celery, Chinese cabbage, Japanese cabbage, cubs, and red cubs. , Chinese cabbage, yellow paprika, sprout cabbage, zucchini, carrot, gobo, renkon, Yamato mana (Yamato vegetables), Chinese cabbage (Yamato vegetables), Uda gold gobo (Yamato vegetables), buds (Yamato vegetables), Akane Katahira (Yamato vegetables) ) Vegetables are preferable.

The food or drink of the present embodiment may be in a form that can be orally ingested, such as a solution, a solid form, or a powder, and is not particularly limited. Specific examples include beverages (lactic acid beverages, fruit juice beverages, soy milk beverages, vegetable beverages, tea beverages, carbonated beverages, nutritional beverages, sports beverages, coffee beverages, soups, alcoholic beverages, etc.), dairy products (yogurt, cheese, butter, etc.) Ice cream, etc.), flour products (bread, noodles, cake mix, etc.), confectionery (chocolate, cookies, candy, caramel, jelly, gum, Japanese confectionery, etc.), fats and oils foods (dressing, mayonnaise, cream, etc.), seasonings (source) , Tomato ketchup, vinegar, flavor seasoning, soup source, etc.), Instant foods (instant noodles, instant soups, miso soup, canned foods, retort foods, etc.), supplements (tablets, syrups, granules, capsules, quick-disintegrating agents) Etc.).

The lactic acid bacterium of the present embodiment is used for various purposes in the same manner as the lactic acid bacterium used in conventional probiotics, and by ingesting it, physiological effects such as intestinal regulation and antibacterial effects can be expected.

If the lactic acid bacteria of the present embodiment will be ingested by humans or animals, there is no strict limits on the amount, the preferred intake is preferably 10 14 ^CFU from 10 6 ^CFU per the viable cell count. Further, the lactic acid bacterium of the present embodiment is preferably continuously ingested, but is not limited to this, and may be ingested at intervals such as every other week or every other day, or may be ingested for a short period of time.

Various nutrients, various vitamins, various minerals, stabilizers such as sweeteners and emulsifiers, thickeners, dietary fiber, flavors and other optional ingredients can be added to the foods and drinks and fermented products.

Nutrients include DHA EPA, acetylglucosamine, catechin, turmeric, propolis, agarisk, β-cryptoxanthin, quercetin, anthocyanins and the like. Examples of various vitamins include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, folic acid, nicotine amide, biotin, and vitamin K. Examples of various minerals include magnesium, calcium, zinc, manganese, iron, sodium, potassium and the like. Sweeteners include sucrose, glucose, fructose, trehalose, xylose, lactose, palatinose, fructose-glucose liquid sugar, malt sugar, fructose, honey and other sugars, sorbitol, xylitol, erythritol, lactitol, palatinit, aspartame, sclarose, stevia, High-sweetness sweeteners such as Acesulfam K can be mentioned. Examples of the acidulant include citric acid, lactic acid, acetic acid, malic acid, tartaric acid, butyric acid and the like. Flavors include orange, citrus, berry, apple, mint, grape, apricot, perilla, lemon, grapefruit, peach, banana, tropical, herb, coffee, and tea. And so on.

In addition, when producing the fermented product or food or drink of the present embodiment, bacteria other than the lactic acid bacteria of the present embodiment can be used in combination. Examples of such fungi include Bifidbacterium spp., Lactobacillus casei, Lactobacillus acidophilus, and other Bifidbacterium spp. Lactobacillus blevis, Lactobacillus kefia, Lactobacillus paracasei, Lactobacillus salivalius, Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus del Burky, Lactobacillus pentosas, Lactobacillus Lactobacillus bacteria such as Plantalum, Lactobacillus bacteria such as Streptococcus thermophilis, Streptococcus salivalius, Lactobacillus lactis, Lactis subspicis lactis, Lactobacillus lactis, Subspicis, Cremoris, etc. Examples include yeasts belonging to the genus Enterococcus, the genus Saccalomyces such as Saccalomyces cervisier and Candida Kefia, and the genus Candida.

Hereinafter, the contents of the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

Example 1
Isolation and culture conditions of lactic acid bacteria of Examples The novel lactic acid bacteria belonging to Lactobacillus plantarum were isolated using the sake lees filtration medium shown below. Add 200 g of sake lees to 800 ml of distilled water, heat over low heat, stir well and bring to a boil to dissolve. It was removed from the heat and cooled to about 50 ° C., filtered with kitchen paper, Brix was adjusted to 5 and 0, autoclaved at 121 ° C. for 15 minutes, and used as a sake lees filtration medium. The fungi attached to the flowers of Yaezakura were brought into contact with a sake lees filtration medium and cultured at 30 ° C. for 24 hours to isolate the grown fungi. After isolation, bacteria that could be cultured in Lactobacillus MRS Broth (Difco) medium autoclaved at 121 ° C. for 15 minutes at 30 ° C. for 24 hours were used in subsequent experiments. Such bacteria formed round, white colonies in Lactobacillus MRS Broth (Difco) medium. Upon microscopic observation, the above-mentioned bacterium was a bacillus, had no motility, and did not form spores. (Not shown). The above bacteria showed positive in the Gram stain test, showed no activity in the catalase activity test, and no gas generation was observed. (Not shown). Further, it has been confirmed that the above-mentioned bacteria grow under anaerobic conditions, but also grow in the presence of air.

Bacterial species identification using 16S rDNA base sequence Bacterial species identification of the above bacteria was performed by the following method based on the 16S rDNA base sequence. The entire length of the 16S rDNA sequence was amplified by the PCR method using the DNA extracted from the cell pellet obtained by centrifuging the culture solution cultured at 30 ° C. for 24 hours using MRS medium as a template, and the nucleotide sequence of the amplified product was determined by Dye Terminater. Determined by law. The base sequence is shown in SEQ ID NO: 1. The obtained nucleotide sequence was searched in a database to identify the bacterial species.

As a result, it was confirmed that the base sequence of 16S rDNA of the lactic acid bacterium was 99.2% identical to the base sequence of 16S rDNA of Lactobacillus plantarum and Lactobacillus pentosas, but Lactobacillus pentosas was D-xylose. However, since the lactic acid bacterium cannot assimilate D-xylose, the lactic acid bacterium was identified as a new strain of lactic acid bacterium belonging to Lactobacillus plantarum and named Lactobacillus plantarum Sakura 68 (hereinafter referred to as Lactobacillus plantarum Sakura 68). (Lactobacillus case of Example 1) was deposited on January 29, 2021 at the Patent Microbial Depositary Center of the Product Evaluation Infrastructure Organization, which has an address in 2-5-8 Kazusa Kazusa, Chiba Prefecture, Japan. The accession number is NITE P-03374.

The saccharification property of the lactic acid bacteria of the examples is shown in Table 1 (fermentation properties of sugar: measured by API50CH).

Example 2
Fermentation test of fruit juice, vegetable juice, and vegetable mixed juice with lactobacillus plantalum (1) Strains Two strains belonging to lactobacillus plantalum as lactic acid bacteria (control group) of Comparative Example 1 YIT0132 strain (FERM BP-11349) (Comparative Example 1) -1), YIT0148 (FERM BP-11350) (Comparative Example 1-2) was used.

(2) Inoculum and culture The lactic acid bacteria of Comparative Examples 1-1 and 1-2 were inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C for 15 minutes, and the mouth was loosened in the presence of air to 37 ° C. The cells were cultured for 16 hours until they were in a steady state.

The lactic acid bacterium of Example 1 (Lactobacillus plantarum Sakura 68) was inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C. for 15 minutes, and the mouth was loosened in the presence of air for 16 hours at 30 ° C. It was cultured until it became a steady state.

(3) Proliferation in fruit juice, vegetable juice, mixed vegetable juice Orange (manufactured by Yakult Headquarters), Orange (manufactured by Kirin Co., Ltd.), Pine (manufactured by Yakult Headquarters), Pine (manufactured by Kirin Co., Ltd.), Apple (manufactured by Yakult Headquarters), Apple (Kirin), 100% grapefruit (Kirin) juice, carrot (Carrot 100, Itoen), tomato (high lycopene, Kagome), tomato (premium, Kagome), tomato (ideal) Tomato, Itoen Co., Ltd.), Tomato (ripe tomato, Delmonte Co., Ltd.), Tomato (salt-free, Delmonte Co., Ltd.) 100% vegetable juice, vegetable mixed juice (Kinari vegetables, Yakult headquarters), vegetables Mixed juice (1 bottle of vegetables per day, made by Yakult Headquarters), vegetable mixed juice (1 day's worth of vegetables, made by Itoensha), vegetable mixed juice (full vegetables, made by Itoensha), vegetable mixed juice (vegetable life, Kagome) (Made by Kirin) and vegetable mixed juice (48 kinds of dark vegetables, manufactured by Kirin) were aseptically dispensed into test tubes, and Comparative Example 1-1 YIT0132 (FERM BP-11349), 1-2 YIT0148 (FERM BP-) 11350) was inoculated with 0.4% volume of the bacterial solution and cultured at 37 ° C. for 48 hours.

The above-mentioned commercially available fruit juice, vegetable juice, and vegetable mixed juice were aseptically dispensed into test tubes, inoculated with a 0.4% volume of the lactic acid bacterium of Example 1, and cultured at 30 ° C. for 48 hours.

(4) Measurement of viable cell count The culture solution was appropriately diluted with a diluted solution (0.1% yeast extract solution) and then applied to an MRS agar plate medium with a spanal system to measure the viable cell count. Table 2 shows the results of examining the proliferative properties of fruit juice, vegetable juice, and mixed vegetable juice. The viable cell count was calculated by performing three different experiments and averaging the obtained values.

(5) Examination of Proliferativeness in Fruit Juice, Vegetable Juice, and Vegetable Mixed Juice The lactic acid bacteria of Example 1 were about 1.7 times as much in all of the fruit juice, vegetable juice, and mixed vegetable juice as compared with the lactic acid bacteria of Comparative Example 2 strains. It was clarified that it showed 4 times the proliferative ability and had excellent proliferative ability.

Example 3
Fermentation experiment of crushed fruit with Lactobacillus plantarum (1) Strain Two strains belonging to Lactobacillus plantarum as lactic acid bacteria (control group) of Comparative Example 1 YIT0132 strain (FERM BP-11349) (Comparative Example 1-1), YIT0148 (FERM BP-11350) (Comparative Example 1-2) was used.

(2) Inoculum and culture The lactic acid bacteria of Comparative Examples 1-1 and 1-2 were inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C for 15 minutes, and the mouth was loosened in the presence of air to 37 ° C. The cells were cultured for 16 hours until they were in a steady state.

The lactic acid bacterium of Example 1 (Lactobacillus plantarum Sakura 68) was inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C. for 15 minutes, and the mouth was loosened in the presence of air for 16 hours at 30 ° C. It was cultured until it became a steady state.

(3) Proliferation in crushed fruit Fruit crushed apple, strawberry, pear, watermelon, melon, kiwi, mango, papaya, and red dragon fruit were aseptically dispensed into test tubes and compared with Comparative Example 1-1. Each of them was inoculated with 0.4% volume of bacterial solution of YIT0132 (FERM BP-11349) and 1-2 YIT0148 (FERM BP-11350), and cultured at 37 ° C. for 48 hours. The same crushed fruit products were aseptically dispensed into test tubes, and each of them was inoculated with a bacterial solution having a volume of 0.4% of the lactic acid bacterium of Example 1 and cultured at 30 ° C. for 48 hours.

(4) Measurement of viable cell count The culture solution was appropriately diluted with a diluted solution (0.1% yeast extract solution), and then applied to an MRS agar plate medium with a spanal system to measure the viable cell count. Table 3 shows the results of examining the proliferative property of crushed fruit. The viable cell count was calculated by performing three different experiments and averaging the obtained values.

(5) Examination of Proliferativeness in Crushed Fruits The lactic acid bacteria of Example 1 showed about 1.6 to 14 times the viable count of all crushed fruits as compared with the two strains of lactic acid bacteria of Comparative Example. From this, it was clarified that it has excellent proliferative ability.

Example 4
Fermentation experiment of crushed vegetables with Lactobacillus plantarum (1) Strains Two strains belonging to Lactobacillus plantarum as lactic acid bacteria (control group) of Comparative Example 1 YIT0132 strain (FERM BP-11349) (Comparative Example 1-1), YIT0148 (FERM BP-11350) (Comparative Example 1-2) was used.

(2) Culture of inoculum The lactic acid bacteria of Comparative Examples 1-1 and 1-2 were inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C for 15 minutes, and the mouth was loosened in the presence of air to 37 ° C. The cells were cultured for 16 hours until they reached a steady state.
The lactic acid bacterium of Example 1 (Lactobacillus plantarum Sakura 68) was inoculated after autoclaving sterilization of Lactobacilli MRS Broth (manufactured by Difco) at 121 ° C. for 15 minutes, and the mouth was loosened in the presence of air for 16 hours at 30 ° C. It was cultured until it became a steady state.

(3) Proliferation in crushed vegetables Tomato, carrot, cabbage, lettuce, cucumber, onion, Naganegi, celery, Chinese cabbage, Japanese mustard spinach, turnip, red turnip, pepper, yellow paprika, broccoli, asparagus, Mizuna, Shungiku, spinach , Komatsuna, bok choy, nanohana, parsley, seri, bean seedling, Brussels sprouts, bitter gourd, zucchini, ginger, garlic, burdock, renkon, creson, mana Yamato (Yamato vegetables), white vegetables (Yamato vegetables), Uda gold gobo (Yamato vegetables) ), Celebrations (Yamato vegetables), and Akane Katahira (Yamato vegetables) are aseptically dispensed into test tubes, and comparative strain example 1-1 YIT 0132 (FERM BP-11349), 1-2 YIT. Each of the bacterial solutions having a volume of 0.4% of 0148 (FERM BP-11350) was inoculated and cultured at 37 ° C. for 48 hours. The same crushed vegetables were aseptically dispensed into test tubes, inoculated with 0.4% volume of the lactic acid bacterium of Example 1, and cultured at 30 ° C. for 48 hours.

(4) Measurement of viable cell count The culture solution was appropriately diluted with a diluted solution (0.1% yeast extract solution), and then applied to an MRS agar plate medium with a spanal system to measure the viable cell count. Table 4 shows the results of examining the proliferative property of crushed vegetables. The viable cell count was calculated by averaging the values obtained by conducting the experiment three times each.

(5) Results The lactic acid bacteria of Example 1 were excellent because they showed about 1.3 to 13 times the viable count of all crushed vegetables as compared with the two strains of lactic acid bacteria of Comparative Example. It was revealed that it has a proliferative ability.

As shown in Tables 1, 2 and 3, the lactic acid bacteria of Example 1 are various as compared with the lactic acid bacteria of Comparative Example 1-1 YIT 0132 (FERM BP-11349) 1-2 (FERM BP-11350). It was clarified that in fruit juice, vegetable juice, mixed vegetable juice, crushed fruit, and crushed vegetable, the number of viable bacteria reached was always high and the growth ability was excellent.

Claims (11)

Lactic acid bacteria that meet the following properties and are entrusted with the entrustment number: NITE P-03374.
The lactic acid bacteria solution cultured until a steady state is reached the number of viable bacteria when cultured for 48 hours at 30 ° C. those 0.4% volume inoculation against plant food is 10 9 ^CFU / ml or more. The lactic acid bacterium according to claim 1, wherein the plant-based food and drink is at least one selected from the group consisting of fruit juice, vegetable juice, vegetable mixed juice, crushed fruit and crushed vegetables. The lactic acid bacterium according to claim 1 or 2, which satisfies at least one of the following properties (a), (b) and (c).
(A) Lactobacillus cultivated in Lactobacilli MRS Broth medium at 30 ° C. for 16 hours at 30 ° C. was added to 100% fruit juice of orange and pine, 100% vegetable juice of tomato and carrot, and 0. 4% volume inoculated reach viable count when cultured for 48 hours at 30 ° C. what was found at both 10 9 ^CFU / ml or more.
(b) Lactobacillus solution cultivated in Lactobacilli MRS Broth medium at 30 ° C. for 16 hours was applied to crushed fruits of strawberry, pear, watermelon, melon, mango, papaya, and red dragon fruit by 0.4 each. % volume reached viable count when cultured for 48 hours at 30 ° C. those inoculated is both 10 ⁹ CFU / ml or more.
(c) Tomato, carrot, cabbage, lettuce, onion, onion, celery, Chinese cabbage, radish, turnip, red turnip, pepper, yellow Paprika, sprout cabbage, zucchini, carrot, burdock, lotus root, mana Yamato (Yamato vegetables), Chinese cabbage (Yamato vegetables), Uda Kingobo (Yamato vegetables), turnips (Shukuri) (Yamato vegetables), Akane Katahira (Yamato vegetables) arrival number of viable bacteria are both 10 ⁹ CFU / ml or more when cultured for 48 hours at 30 ° C. those 0.4% volume inoculated respectively vegetable crushed. The composition obtained by fermentation with the lactic acid bacterium according to any one of claims 1 to 3. A composition comprising the lactic acid bacterium according to any one of claims 1 to 3 and a food material. The composition according to claim 5 , wherein the food material is a material derived from a plant. The composition according to claim 6 , wherein the material derived from the plant is leaves, roots, stems, flowers, buds, fruits, extracts thereof, crushed products thereof, and / or a combination thereof. The composition according to claim 6 , wherein the plant-derived material is a fruit, a vegetable, an extract thereof, a crushed product thereof, and / or a combination thereof. The composition according to any one of claims 4 to 8 , wherein the composition is a fermented product. A food or drink containing the lactic acid bacterium according to any one of claims 1 to 3 or the composition according to any one of claims 4 to 9. A method for producing a composition, which comprises inoculating a food material with the lactic acid bacterium according to any one of claims 1 to 3 and fermenting the food material.