Novel lactic acid bacteria belonging to three species of lactiplantibacillus plantarum isolated from plum fruit, wisteria flower, and persimmon fruit, and use thereof

Abstract

To solve the following problem that: to date, the only case in which tofu itself has been fermented is a product obtained by fermenting tofu by immersing the tofu in a fermentation liquid in which koji and lactic acid bacteria are mixed.SOLUTION: The purpose of the present invention is to provide lactic acid bacteria satisfying the following properties: a lactic acid bacterial liquid containing lactic acid bacteria cultured until reaching a stationary state, when inoculated into tofu at 1%(V / W) and cultured at 30°C for 24 hours, attains a bacterial count of 3×109 CFU / g or more.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to novel lactic acid bacteria belonging to Lactiplantibacillus plantarum that are excellent at fermenting crushed firm tofu and crushed silken tofu, foods and beverages containing the lactic acid bacteria, fermented products obtained by the lactic acid bacteria, foods and beverages containing the fermented products, and methods for producing fermented products using the lactic acid bacteria. [Background technology]

[0002] Japan today is said to be living in an age of plenty, where not only are foods from all over the world available, but people can also eat cuisine from all over the world.

[0003] The food industry produces a huge amount of food every day to meet the demands of many people, resulting in excess food being wasted without being eaten.

[0004] These include firm tofu and silken tofu, which are made by adding bittern to soy milk and solidifying it. Tofu is said to have been introduced from China during the Nara period, and became popular among the general public during the Edo period, and has continued to be popular to this day.

[0005] Nowadays, it is no exaggeration to say that it is a national dish of Japan, and it is an indispensable ingredient in cold tofu in the summer and in hot pot in the winter.

[0006] To satisfy the dietary needs of the many people in Japan, major tofu manufacturers produce and sell large quantities of tofu every day. As a result, not only are there large amounts of unsold tofu, but because tofu is soft and breaks easily, a large amount of tofu becomes unsold during transportation.

[0007] Tofu that cannot be sold in this way has a shelf life of only a few days and cannot be used for other products, so it is discarded every day.

[0008] Major tofu manufacturers are throwing away tens of thousands of blocks of tofu per day.

[0009] Tofu is discarded daily in this way, and to date there has been almost no development into using it in other new foods.

[0010] This problem could be solved if discarded tofu could be made to last longer through fermentation and then repurposed into a new product with added value through fermentation.

[0011] Tofu is originally made by solidifying soy milk with bittern, so it may be possible to ferment it using plant-based lactic acid bacteria that are excellent at fermenting soy milk.

[0012] There are two main types of tofu: firm tofu and silken tofu. Silken tofu is made by adding bittern to soy milk and solidifying it, but firm tofu is made by first breaking up the solidified tofu, then placing a cotton cloth over it and pressing it down. This causes the moisture to escape and the nutrients to be compressed, but at the same time, water-soluble components are also lost.

[0013] The nutritional value of firm tofu is nearly 1.3 times higher than that of silken tofu, but silken tofu contains more aqueous vitamin B complex, potassium, and carbohydrates than firm tofu, and nearly 2.5 times more carbohydrates than firm tofu.

[0014] For these reasons, soy milk, firm tofu, and silken tofu can be said to be different foods in terms of properties and nutritional aspects.

[0015] Yogurt-like products made by fermenting soy milk have been commercialized, but yogurt and cheese-like products have also been produced by precipitating soy milk with acids or fermenting it using yeast or koji (Patent Document 1). However, there are no commercially available cheese-like foods made by fermenting soy milk with lactic acid bacteria, in the same way as cheese made by conventional lactic acid fermentation of milk. [Prior art documents] [Patent documents]

[0016] [Patent Document 1] Patent Publication No. 2008-067680 Summary of the Invention [Problem to be solved by the invention]

[0017] However, to date, the only products that have fermented tofu itself are those that are soaked in a fermentation liquid containing a mixture of koji and lactic acid bacteria (for example, Patent Document 1). Lactic acid bacteria are not used alone to ferment the tofu, and since the tofu is simply soaked, the tofu is not fermented uniformly. Furthermore, the number of lactic acid bacteria in the fermented tofu has not been investigated, so it is unclear whether the number of bacteria required to obtain the effects of probiotics has been achieved. [Means for solving the problem]

[0018] Therefore, one object of the present invention is to provide a novel lactic acid bacterium belonging to the genus Lactiplastibacillus plantarum that has excellent fermentation ability, achieving a high viable cell count even when plant components such as firm tofu or silken tofu are used as fermentation substrates, and a food or drink containing the same. Another object of the present invention is to provide lactic acid bacteria-fermented tofu obtained by fermenting tofu using only lactic acid bacteria in the absence of koji mold, and a method for producing the same.

[0019] As a result of extensive research conducted by the present inventors to solve the above problems, it was found that three types of lactic acid bacteria belonging to Lactiplantibacillus plantarum isolated from plums, wisteria flowers, and persimmons can achieve a viable cell count of 3 x 10, which is sufficient to exert probiotic functions, even when crushed firm tofu is used as the fermentation substrate. 9 Even when crushed silken tofu is used as the fermentation substrate, the number of viable bacteria reached is 3 x 10, which is sufficient to exert probiotic functions. 9 The inventors have found that the CFU / g or more is higher than that of the control, and have completed the present invention.

[0020] [1] That is, the object of the present invention is to The present invention provides lactic acid bacteria belonging to Lactiplantibacillus plantarum, which satisfy the following properties and are entrusted under accession numbers NITE AP-04123, NITE AP-04124, and NITE AP-04125: The lactic acid bacteria solution containing lactic acid bacteria cultured in Lactobacilli MRS Broth medium at 30°C for 16 hours until the bacteria reached a steady state was inoculated into tofu at 1% volume (V / W) and cultured at 30°C for 24 hours. The viable cell count reached was 3 x 10 9 CFU / g or more.

[0021] By using this lactic acid bacterium, tofu can be fermented.

[0022] [2] Another object of the present invention is to provide a composition obtained by fermenting a food material with the lactic acid bacterium described in [1].

[0023] [3] Another object of the present invention is to provide a composition comprising the lactic acid bacteria according to [1] and a food material.

[0024] [4] In the composition according to [2] or [3], the food material may be a material derived from a plant.

[0025] [5] In the composition described in [4], the material derived from the plant may be the leaves, roots, stems, fruits, extracts thereof, crushed products thereof, processed products thereof, and / or combinations thereof.

[0026] [6] [5] The composition according to [5], The plant may be soybean; The extract may be soy milk; The processed product may be tofu.

[0027] [7] In the composition according to [2] or [3], the composition may be a fermented product.

[0028] [8] Another object of the present invention is to provide a food or drink containing the lactic acid bacteria according to [1] or the composition according to [2] or [3].

[0029] [9] Another object of the present invention is to provide a method for producing a composition, which comprises inoculating a food material with the lactic acid bacteria described in [1] and fermenting the food material.

[0030]

[10] Another object of the present invention is to provide lactic acid bacteria-fermented tofu, which is obtained by inoculating tofu with only the lactic acid bacteria described in [1] as the fermenting bacteria, and fermenting the tofu.

[0031]

[11] In the lactic acid bacteria-fermented tofu described in

[10] , the lactic acid bacteria may be at least one of the three types of lactic acid bacteria described in [1].

[0032]

[12] Another object of the present invention is to provide crushing the tofu to obtain pulverized tofu; a step of inoculating the crushed tofu with only the lactic acid bacteria described in [1] as fermenting bacteria; fermenting the crushed tofu inoculated with the lactic acid bacteria; The present invention provides a method for producing lactic acid bacteria-fermented tofu, comprising the steps of:

[0033]

[13] In the method for producing lactic acid bacteria-fermented tofu described in

[12] , the lactic acid bacteria may be at least one of the three types of lactic acid bacteria described in [1]. [Effects of the Invention]

[0034] The lactic acid bacteria of the present invention can ferment a variety of plants and can be used to prepare foods and beverages, such as probiotic beverages, based on a variety of fermentation substrates. [Brief explanation of the drawings]

[0035] [Figure 1] Figure 1 shows a radar chart of the sourness (mV), bitterness (mV), astringency (mV), umami (mV), bitterness (mV), astringency (mV), umami body (mV), and sweetness (mV) of silken tofu measured using a taste sensor. [Figure 2] Figure 2 shows a radar chart of the sourness (mV), bitterness (mV), astringency (mV), umami (mV), bitterness (mV), astringency (mV), umami body (mV), and sweetness (mV) of firm tofu measured using a taste sensor. [Figure 3] 1 shows photographs of the external appearance and internal condition of the dry-ripened lactic acid bacteria-fermented tofu cheese-like food product of Example 1-1. [Figure 4] 1 shows photographs of the external appearance and internal condition of the dry-ripened lactic acid bacteria-fermented tofu cheese-like food product of Example 1-2. [Figure 5] Photographs of the appearance and internal condition of the dry-ripened lactic acid bacteria-fermented tofu cheese-like food product of Examples 1-3 are shown. DETAILED DESCRIPTION OF THE INVENTION

[0036] definition For convenience, certain terms used in this application are collected here. Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0037] Although the numerical ranges and parameters set forth in the present invention are approximate, the numerical values ​​set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently contains certain errors necessarily resulting 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 within an acceptable standard error as considered by one of ordinary skill in the art.

[0038] "Tofu" as used herein includes firm tofu and silken tofu.

[0039] The term "fermented product" as used herein means a product that has been fermented using lactic acid bacteria, and includes products that have been fermented using fermentation substrates such as food materials as raw materials.

[0040] In this example, "Thickened Cotton Tofu" (manufactured by Takano Foods Co., Ltd.) was used as the firm tofu. In this example, the silken tofu used was "Kokushitate Kinu" (manufactured by Takano Foods Co., Ltd.).

[0041] Hereinafter, embodiments of the present invention will be described. The following embodiments are merely examples, and the scope of the present invention is not limited to the following embodiments. Note that similar content will not be described again to avoid repetition.

[0042] lactic acid bacteria The lactic acid bacteria of this embodiment satisfy the following properties. A lactic acid bacteria solution containing lactic acid bacteria cultured until it reached a steady state was inoculated into tofu at 1% (V / W) and cultured at 30°C for 24 hours, reaching a bacterial count of 3 x 10 9 CFU / g or more.

[0043] In some embodiments, the tofu includes firm tofu and silken tofu.

[0044] The number of viable bacteria reached was 3.0 x 10 9 CFU / g or more, 3.5 x 109 CFU / g or more, 4.0 x 10 9 CFU / gl or more, 4.5 × 10 9 CFU / g or more, 5.0 × 10 9 CFU / g or more, 5.5 x 10 9 CFU / g or more, 6.0 x 10 9 CFU / g or more, 6.5 x 10 9 CFU / g or more, 7.0 × 10 9 CFU / g or more, 7.5 x 10 9 CFU / g or more, 8.0 × 10 9 CFU / g or more, 8.5 x 10 9 CFU / g or more, 9.0 x 10 9 CFU / g or more, 9.5 x 10 9 It may be CFU / g.

[0045] The lactic acid bacteria liquid may be obtained by culturing Lactobacilli in MRS Broth medium, for example, at 30°C for 16 hours.

[0046] In one embodiment, the tofu is crushed tofu. The crushed tofu is prepared by processing 200 g of firm tofu in a juicer until creamy, then weighing 25 g portions into jars with lids, putting the jars in the lids, placing them in an incubator, setting the temperature to 80°C, and sterilizing for 1 hour after the temperature reaches 80°C.

[0047] In this specification, the term "achieved viable cell count" refers to the viable cell count measured after culturing a seed culture in Lactobacilli MRS Broth (Difco) medium, which has been autoclaved at 121°C for 15 minutes, for 16 hours at 30 or 37°C until a steady state is reached, followed by inoculating a 1% volume of the preculture solution into a fermentation substrate of a food material (such as crushed tofu), followed by culturing for 24 hours at 30°C. The viable cell count is measured using MRS agar plates. The unit of viable cell count, CFU / g, indicates the colony-forming ability of 1 g of the culture medium when inoculated onto a culture plate.

[0048] In one embodiment, the lactic acid bacteria belong to the genus Lactobacillus. In another embodiment, the lactic acid bacteria belong to Lactiprantibacillus plantarum. In yet another embodiment, the lactic acid bacteria belong to Super Probiotics-1, Super Probiotics-2, or Super Probiotics-3.

[0049] In yet another embodiment, the lactic acid bacteria are those deposited under accession numbers NITE AP-04123, NITE AP-04124, and NITE AP-04125.

[0050] In one embodiment, the lactic acid bacteria also include mutant strains that can grow under the above conditions.

[0051] To achieve the physiological effects of the lactic acid bacteria of the present invention, they can be used in conventional pharmaceutical formulations, for example, by mixing them with solid or liquid non-toxic pharmaceutical carriers. Examples of such formulations include solid formulations such as tablets, granules, powders, and capsules; liquid formulations such as solutions, suspensions, and emulsions; and lyophilized formulations. These formulations can be prepared using conventional pharmaceutical techniques. Examples of such non-toxic pharmaceutical carriers include fatty acid glycerides, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, glucose, lactose, sucrose, starch, mannitol, dextrin, amino acids, gelatin, albumin, water, and physiological saline. Conventional additives such as stabilizers, humectants, emulsifiers, binders, isotonicity agents, and excipients can also be added as needed.

[0052] Furthermore, the lactic acid bacteria of the present invention can be incorporated into foods and beverages in any form, such as solid or liquid, in addition to the formulations described above. When incorporated into foods and beverages, they may be incorporated as is or together with various nutritional components. Specifically, when incorporating the lactic acid bacteria of the present invention into foods and beverages, additives that can be used in foods and beverages are used appropriately, and the lactic acid bacteria can be formed into edible forms, such as granules, tablets, capsules, pastes, etc., using conventional means. Examples of foods and beverages include processed meat foods such as ham and sausage, processed seafood foods such as kamaboko and chikuwa, bread, confectionery, butter, yogurt, cheese, and powdered milk, as well as beverages such as water, fruit juice, milk, soft drinks, and tea. Incidentally, animal feed is also included in foods and beverages.

[0053] In this embodiment, a composition obtained by fermenting a food material with the lactic acid bacteria is also provided. In this embodiment, a composition comprising the lactic acid bacteria and a food material is also provided. The food material may be a plant-derived material. 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, mushrooms, seaweed, extracts thereof, crushed products thereof, and / or combinations thereof. In one embodiment, the plant is soybean. In one embodiment, the extract is soy milk. In one embodiment, the processed product is tofu.

[0054] In this embodiment, there is also provided a food or drink containing the lactic acid bacteria or the composition. In this embodiment, there is also provided a method for producing a composition, which comprises inoculating a food material with the lactic acid bacteria and fermenting the food material.

[0055] In this embodiment, lactobacillus-fermented tofu is also provided, which is obtained by inoculating tofu with only the above-mentioned lactobacillus as the fermenting bacterium and fermenting it. The lactobacillus may be at least one of the above-mentioned three types of lactobacillus (lactobacillus deposited under accession numbers NITE AP-04123, NITE AP-04124, and NITE AP-04125).

[0056] Fermented products obtained by fermenting food materials with the lactic acid bacteria of the present invention, foods and drinks containing the fermented products, and foods and drinks containing the lactic acid bacteria of the present invention The composition of this embodiment may be a fermented product. The fermented product of this embodiment is obtained by fermenting a food material with the lactic acid bacteria of the present invention. Furthermore, the food or drink of this embodiment includes the fermented product. In this embodiment, the food material is not particularly limited as long as it is usable for food, and examples thereof include plant leaves, roots, stems, flowers, buds, fruits, mushrooms, seaweed, extracts thereof, and crushed products thereof. In particular, those that can be fermented by the lactic acid bacteria of the present invention are preferred, such as firm tofu and silken tofu. Lactic acid bacteria-fermented firm tofu can be obtained by first inoculating and culturing crushed firm tofu that has been sterilized under conditions suitable for the crushed firm tofu to be used with the lactic acid bacteria of the present invention, alone or together with other microorganisms. In the case of silken tofu, lactic acid bacteria-fermented silken tofu can also be obtained by a similar method.

[0057] These fermented products can be mixed with unfermented fruit juice, unfermented vegetable juice, crushed unfermented fruit, crushed unfermented vegetable, alcohol, etc., and various nutrients, vitamins, flavors, etc. can be added to produce final products. These preferably contain the lactic acid bacteria of the present invention in a live state.

[0058] The food and drink of this embodiment may be in any form that can be orally ingested, such as a solution, solid, powder, etc., and is not particularly limited. Specific examples include beverages (lactic acid beverages, fruit juice beverages, soy milk beverages, vegetable beverages, tea beverages, carbonated beverages, energy drinks, sports drinks, coffee beverages, soups, alcoholic beverages, etc.), dairy products (yogurt, cheese, butter, ice cream, etc.), flour products (bread, noodles, cake mixes, etc.), sweets (chocolate, cookies, candy, caramel, jelly, gum, Japanese sweets, etc.), oily foods (dressings, mayonnaise, cream, etc.), seasonings (sauces, tomato ketchup, vinegar, flavor seasonings, soup bases, etc.), instant foods (instant noodles, instant soups, miso soup, canned foods, retort foods, etc.), and supplements (tablets, syrups, granules, capsules, rapid-disintegrating tablets, etc.).

[0059] The lactic acid bacteria of this embodiment can be used for various purposes in the same way as lactic acid bacteria used in conventional probiotics, and by ingesting them, physiological effects such as intestinal regulation and antibacterial action can be expected.

[0060] When humans or animals ingest the lactic acid bacteria of this embodiment, there is no strict limit to the amount, but the preferred amount is 10 viable bacteria count per serving. 6 CFU to 10 14 CFU is preferred. Furthermore, although it is preferred that the lactic acid bacteria of this embodiment be taken continuously, this is not limiting, and the lactic acid bacteria may be taken at intervals such as every other week or every other day, or for a short period of time.

[0061] The above-mentioned foods, beverages and fermented products may contain optional ingredients such as various nutrients, various vitamins, various minerals, sweeteners, stabilizers such as emulsifiers, thickeners, dietary fiber, flavors and the like.

[0062] Nutrients include DHA, EPA, acetylglucosamine, catechin, turmeric, propolis, agaric, β-cryptoxanthin, quercetin, anthocyanins, etc. Vitamins include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, folic acid, nicotinamide, biotin, and vitamin K. Minerals include magnesium, calcium, zinc, manganese, iron, sodium, and potassium. Sweeteners include sugars such as sucrose, glucose, fructose, trehalose, xylose, lactose, palatinose, high-fructose corn syrup, maltose, fructose, and honey, as well as high-intensity sweeteners such as sorbitol, xylitol, erythritol, lactitol, palatinit, aspartame, sucralose, stevia, and acesulfame K. Examples of acidulants include citric acid, lactic acid, acetic acid, malic acid, tartaric acid, butyric acid, etc. Examples of flavors include orange, citrus, berry, apple, mint, grape, apricot, shiso, lemon, grapefruit, peach, banana, tropical, herb, coffee, tea, etc.

[0063] Furthermore, when producing the fermented product or food or drink of this embodiment, bacteria other than the lactic acid bacteria of this embodiment can be used in combination. Examples of such bacteria include Bifidobacterium bacteria such as Bifidobacterium, bifidum, Bifidobacterium, lactis, and Bifidobacterium, longum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus kefir, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus delbrueckii, and Lactobacillus penicillin. Examples of such bacteria include Lactobacillus bacteria such as Lactobacillus tosus and Lactobacillus plantarum, Streptococcus bacteria such as Streptococcus thermophilis and Streptococcus salivarius, Lactococcus bacteria such as Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris, Enterococcus bacteria such as Enterococcus faecalis, and yeasts belonging to the Saccharomyces cerevisiae and Candida kefir, and the Candida genus.

[0064] Manufacturing method for lactic acid bacteria fermented tofu The method for producing lactobacillus-fermented tofu according to this embodiment includes the steps of crushing tofu to obtain crushed tofu, inoculating the crushed tofu with the lactobacillus as the only fermenting bacterium, and fermenting the crushed tofu inoculated with the lactobacillus. The lactobacillus may be at least one of the three lactobacillus species deposited under accession numbers NITE AP-04123, NITE AP-04124, and NITE AP-04125.

[0065] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. [Example]

[0066] Example 1 Isolation and culture conditions of lactic acid bacteria in the examples Super Probiotic-1, belonging to Lactobacillus plantarum, was isolated from plum fruit, Super Probiotic-2 from wisteria flowers, and Super Probiotic-3 from persimmon fruit. After isolation, the bacteria that could be cultured in Lactobacilli MRS Broth (Difco) medium autoclaved at 121°C for 15 minutes and then at 30°C for 24 hours were used in subsequent experiments.

[0067] These bacteria formed white circular colonies on Lactobacilli MRS agar plates. Microscopic observation revealed that the bacteria were rod-shaped, non-motile, and did not form spores (not shown). The bacteria showed a positive Gram staining test, showed no activity in a catalase activity test, and did not produce gas (not shown). The bacteria grow under anaerobic conditions, but have also been confirmed to grow in the presence of air.

[0068] Bacterial species identification using 16S rDNA sequences The species of the above bacteria were identified based on the 16S rDNA sequence as follows. The culture solution was cultured in MRS medium at 30°C for 24 hours, and then centrifuged to obtain a bacterial pellet. DNA extracted from the pellet was used as a template to amplify the full length of the 16S rDNA sequence by PCR. The nucleotide sequences of the amplified products were determined by the dye terminator method. The nucleotide sequences are shown in SEQ ID NO: 1 (plum lactic acid bacteria), SEQ ID NO: 2 (wisteria lactic acid bacteria), and SEQ ID NO: 3 (persimmon lactic acid bacteria). The obtained nucleotide sequences were searched against a database to identify the species.

[0069] As a result, the 16S rDNA sequences of the above lactic acid bacteria and the plum lactic acid bacteria showed 99.7% homology with the 16S rDNA of Lactipranchibacillus plantarum and Lactobacillus pentosus, the 16S rDNA sequence of the wisteria lactic acid bacteria showed 99.25% homology with the 16S rDNA of Lactipranchibacillus plantarum, and the 16S rDNA sequence of the persimmon lactic acid bacteria showed 99.25% homology with the 16S rDNA of Lactipranchibacillus plantarum. Although they showed 99.9% homology, Lactobacillus pentosus was able to utilize D-xylose, whereas none of the three lactic acid bacteria could. Furthermore, because these three lactic acid bacteria exhibited different assimilation abilities, they were identified as new strains of lactic acid bacteria belonging to Lactiplantibacillus plantarum, named Super Probiotic-1, Super Probiotic-2, and Super Probiotic-3 (hereinafter referred to as the lactic acid bacteria of Examples 1-1, 1-2, and 1-3, respectively), and were deposited at the National Institute for Product Evaluation and Research (NIER) Patent Microorganisms Depositary, 2-5-8, Kisarazu Kazusa Kamatari, Chiba Prefecture, Japan, on June 5, 2024. The accession numbers for Super Probiotic-1, Super Probiotic-2, and Super Probiotic-3 are NITE AP-04123, NITE AP-04124, and NITE AP-04125, respectively.

[0070] The sugar utilization properties of the lactic acid bacteria of Examples 1-1, 1-2 and 1-3 are shown in Tables 1, 2 and 3, respectively (sugar fermentation properties: measured by API50CH).

[0071] [Table 1]

[0072] [Table 2]

[0073] [Table 3]

[0074] Example 2 Fermentation experiment of crushed firm tofu with lactiprancid Bacillus plantarum (1) Strains Seven strains of Lactobacillus plantarum were used as comparative lactic acid bacteria (control group): strain 3070 (Comparative Example 1-1), strain 3074 (Comparative Example 1-2), strain 12006 (Comparative Example 1-3), strain 101973 (Comparative Example 1-4), strain 101975 (Comparative Example 1-5), strain 101977 (Comparative Example 1-6), and strain 109604 (Comparative Example 1-7).

[0075] (2) Seed culture and cultivation The lactic acid bacteria of Comparative Examples 1-1 to 1-7 were inoculated into Lactobacilli MRS Broth (manufactured by Difco) after autoclaving at 121°C for 15 minutes, and cultured at 30°C for 16 hours until a steady state was reached.

[0076] The lactic acid bacteria Super Probiotics-1, Super Probiotics-2, and Super Probiotics-3 in Examples 1-1, 1-2, and 1-3 were inoculated into Lactobacilli MRS Broth (manufactured by Difco) after autoclaving at 121°C for 15 minutes, and then cultured at 30°C for 16 hours until a steady state was reached.

[0077] (3) Growth in crushed firm tofu A sample containing 25 g of crushed firm tofu prepared from thickened firm tofu (manufactured by Takano Foods Co., Ltd.) was sterilized, and then inoculated with 1% volume (V / W) of the bacterial solution of Comparative Examples 1-1 to 1-7, and cultured at 30°C for 24 hours.

[0078] A sterilized sample containing 25 g of crushed firm tofu was inoculated with a 1% volume (V / W) bacterial solution of each of the lactic acid bacteria of Examples 1-1, 1-2, and 1-3, and cultured at 30°C for 24 hours.

[0079] Measurement of viable bacterial count A sterilized dilution solution (0.1% yeast extract solution) of the culture medium was applied to MRS agar plates using a serial dilution method, and the viable cell count was measured. Growth on crushed firm tofu was examined, and the results are shown in Table 4. The viable cell count was calculated by averaging the values ​​obtained from three separate experiments.

[0080] [Table 4]

[0081] Examination of proliferation in crushed firm tofu The lactic acid bacteria of Examples 1-1, 1-2, and 1-3 showed growth ability approximately 1.47 to 20.3 times higher than that of the lactic acid bacteria of Comparative Examples 1-1 to 1-7, demonstrating superior growth ability.

[0082] Example 3 Fermentation experiment of crushed silken tofu with lactiprancid Bacillus plantarum (1) Strains Seven strains of Lactobacillus plantarum were used as comparative lactic acid bacteria (control group): strain 3070 (Comparative Example 1-1), strain 3074 (Comparative Example 1-2), strain 12006 (Comparative Example 1-3), strain 101973 (Comparative Example 1-4), strain 101975 (Comparative Example 1-5), strain 101977 (Comparative Example 1-6), and strain 109604 (Comparative Example 1-7).

[0083] (2) Seed culture and cultivation The lactic acid bacteria of Comparative Examples 1-1 to 1-7 were inoculated into Lactobacilli MRS Broth (manufactured by Difco) after autoclaving at 121°C for 15 minutes, and cultured at 30°C for 16 hours until a steady state was reached.

[0084] The lactic acid bacteria Super Probiotics-1, Super Probiotics-2, and Super Probiotics-3 in Examples 1-1, 1-2, and 1-3 were inoculated into Lactobacilli MRS Broth (manufactured by Difco) after autoclaving at 121°C for 15 minutes, and then cultured at 30°C for 16 hours until a steady state was reached.

[0085] (3) Growth in crushed silken tofu A sample containing 25 g of crushed silken tofu prepared from thick silk (manufactured by Takano Foods Co., Ltd.) was sterilized, and then inoculated with 1% volume (V / W) of the bacterial solution of Comparative Examples 1-1 to 1-7, and cultured at 30°C for 24 hours.

[0086] A sterilized sample containing 25 g of the above-mentioned crushed silken tofu was inoculated with a 1% volume (V / W) bacterial solution of each of the lactic acid bacteria of Examples 1-1, 1-2, and 1-3, and cultured at 30°C for 24 hours.

[0087] Measurement of viable bacterial count A sterilized dilution solution (0.1% yeast extract solution) of the culture medium was applied to MRS agar plates using a serial dilution method, and the viable cell count was measured. Growth on crushed silken tofu was examined, and the results are shown in Table 5. The viable cell count was calculated by averaging the values ​​obtained from three separate experiments.

[0088] [Table 5]

[0089] Examination of growth in crushed silken tofu The lactic acid bacteria of Example 1 exhibited a proliferation ability that was approximately 1.31 to 59.4 times greater than that of the lactic acid bacteria of Comparative Examples 1-1 to 1-7, demonstrating their superior proliferation ability.

[0090] Example 4 Preparation of lactic acid bacteria-fermented tofu cheese-like food using Examples 1-1, 1-2, and 1-3 as starters (1) Using thickened cotton (Takano Foods Co., Ltd.) and thickened silk (Takano Foods Co., Ltd.), crush the material in a juicer until it becomes creamy, measure out 200g into Tupperware (preparing two samples of each), cover, and place in an incubator. Set the incubator to 80°C, and once the temperature inside the cabinet reaches 80°C, sterilize and adjust for one hour.

[0091] (2) The sterilized samples were inoculated with 1% volume (V / W) of a bacterial solution of each of the lactic acid bacteria of Examples 1-1, 1-2, and 1-3, and fermented at 30°C for 24 hours.

[0092] (3) Two fermented samples, each 200 g, were mixed together, and 40 g each from the crushed firm tofu fermented in Examples 1-1, 1-2, and 1-3, 40 g each from the crushed silken tofu fermented in Examples 1-1 and 1-2, and 80 g from the crushed silken tofu fermented in Example 1-3 were used for measurement using a taste sensor (TS-5000Z, Intelligent Sensor Technology, Inc.).

[0093] The test solution for the taste sensor was obtained by the following procedure. 40g of lactic acid bacteria-fermented tofu was weighed into a bowl and 160g of ultrapure water was added. The mixture was mixed with a hand mixer for 1 minute, and any remaining foam was removed with a spoon. The mixture was transferred to a 250mL centrifuge tube and centrifuged at 5000 rpm for 10 minutes. The aqueous layer was filtered through a nonwoven fabric, and the filtrate was used as the test solution.

[0094] The umami test solution for testing umami was obtained by the following procedure. 330 g of 1 / 3 standard solution was prepared (110 g of standard solution + 220 g of ultrapure water diluted by weight). 1.5 mL of the above test solution was placed in a Falcon tube, and 46.5 mL of the 1 / 3 standard solution was added. The homogenized solution was used as the umami test solution.

[0095] The conditions for the taste sensor are shown in Tables 6 and 7.

[0096] [Table 6]

[0097] [Table 7]

[0098] The average relative values ​​of the taste sensors are shown in Tables 8 and 9, and radar charts based on these average values ​​(excluding saltiness) are shown in Figure 1 (silken tofu) and Figure 2 (firm tofu).

[0099] [Table 8]

[0100] [Table 9]

[0101] The remaining sample used in the taste sensor was wrapped in gauze, placed in a wooden frame, and dehydrated for one day with a weight of approximately 2 kg placed on top. After dehydration, it was immersed in 15% salt water for 30 minutes. It was then placed in a refrigerator at 4°C for 10 days to dry and mature.

[0102] The weights of each sample before and after dehydration and after dry-aging are shown in Table 10. Photographs of the external appearance and internal condition of each sample (lactic acid bacteria-fermented tofu cheese-like food) after dry-aging are shown in Figure 3 (Example 1-1), Figure 4 (Example 1-2), and Figure 5 (Example 1-3).

[0103] [Table 10]

[0104] Sensory evaluation Using the sensory evaluation index (p. 38) from "Development of a method for evaluating and designing the palatability of natural cheese based on food sensory engineering" (Doctoral dissertation by Morita Airi, University of Tokyo), a sensory evaluation was conducted on 15 men and women (aged from their 20s to 80s). Evaluations were conducted on five items: appearance, aroma, flavor, taste, and texture. Multiple answers were allowed for each item. Responses to items that did not apply were not tallied. The results are shown in Tables 11 to 16.

[0105] [Table 11]

[0106] [Table 12]

[0107] [Table 13]

[0108] [Table 14]

[0109] [Table 15]

[0110] [Table 16]

[0111] The sensory evaluation of these 15 people confirmed that the lactobacillus-fermented tofu cheese-like food produced changes color to yellow with maturation, just like cheese made from milk by lactobacillus fermentation. It also confirmed that the texture becomes creamy or sticky, and the taste and aroma resemble those of cheese made from milk by lactobacillus fermentation.

Claims

1. Lactic acid bacteria belonging to Lactiplantibacillus plantarum satisfying the following properties and deposited under accession numbers NITE AP-04123, NITE AP-04124 and NITE AP-04125. A lactic acid bacteria solution containing lactic acid bacteria cultured in Lactobacillus MRS Broth medium at 30°C for 16 hours until a steady state was reached was inoculated into tofu at 1% volume (V / W) and cultured at 30°C for 24 hours. The number of viable bacteria reached was 3 x 10 ９ CFU / g or more.

2. A composition obtained by fermenting a food material with the lactic acid bacteria of claim 1.

3. A composition comprising the lactic acid bacteria of claim 1 and a food material.

4. The composition according to claim 2 or 3, wherein the food material is a material derived from a plant.

5. 5. The composition according to claim 4, wherein the plant-derived material is plant leaves, roots, stems, fruits, extracts thereof, crushed products thereof, processed products thereof and / or combinations thereof.

6. the plant is soybean; the extract is soy milk, The processed product is tofu. The composition of claim 5.

7. The composition of claim 2 or 3, wherein the composition is a fermented product.

8. A food or drink comprising the lactic acid bacteria of claim 1 or the composition of claim 2 or 3.

9. A method for producing a composition, comprising inoculating a food material with the lactic acid bacterium according to claim 1 and fermenting the food material.

10. 10. Lactic acid bacteria-fermented tofu, obtained by inoculating tofu with only the lactic acid bacteria according to claim 1 as the fermenting bacteria, and fermenting the tofu.

11. The lactic acid bacteria-fermented tofu according to claim 10, wherein the lactic acid bacteria is at least one of the three types of lactic acid bacteria according to claim 1.

12. crushing the tofu to obtain pulverized tofu; a step of inoculating the tofu crushed material with only the lactic acid bacteria according to claim 1 as fermenting bacteria; and fermenting the crushed tofu inoculated with the lactic acid bacteria; A method for producing lactic acid bacteria-fermented tofu, comprising:

13. The method for producing lactic acid bacteria-fermented tofu according to claim 12, wherein the lactic acid bacteria is at least one of the three types of lactic acid bacteria according to claim 1.

Images