JP2015198638A - Bifidobacteria fermentation method and proliferation method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a bifidobacteria fermentation method and proliferation method that promote the fermentation process of bifidobacteria and improve the number of viable remaining bacteria of bifidobacteria.SOLUTION: The present invention relates to the bifidobacteria fermentation method and proliferation method that, by performing the fermentation treatment with bifidobacteria using a composition containing a pumpkin processed product, it is found that the proliferation of bifidobacteria is substantially promoted, and a fermentation treatment composition is obtained, in which a viable cell count is large, thereby, promoting the fermentation treatment of bifidobacteria and improving the number of viable remaining bacteria of bifidobacteria, wherein the composition containing the pumpkin treated product consists of 30 mass% or more of a pumpkin crushed material or 16 mass% or more of pumpkin juice based on the total amount of the composition.

Description

  The present invention relates to a bifidobacteria fermentation method or a bifidobacteria growth method that promotes the growth of bifidobacteria and increases the number of surviving bacteria of the bifidobacteria. More specifically, the present invention relates to a fermentation method using a pumpkin crushed or pumpkin juice of bifidobacteria and a method of growing bifidobacteria in enteric bacteria.

In a situation where consumers are interested in increasing awareness of food and improving health and preventing illness through daily eating habits, “improving intestinal flora balance. Probiotics, which are “live bacteria additives that have beneficial effects on animals”, are attracting attention as a means for maintaining and improving health and the like from a viewpoint different from the enhancement of intake of various nutrients. The probiotic conditions are: “It is a microorganism that is safe and constitutes the intestinal flora, has survival resistance to digestive juices such as gastric juice, is easy to settle in the intestinal tract, and has useful properties. One of the important factors for effective use of probiotics is that oral intake of a certain number or more of living microorganisms is one of the important factors. It has become. Lactobacillus genus bacteria, Enterococcus genus bacteria and Bifidobacterium genus bacteria (hereinafter referred to as “Bifidobacterium”) are known as representative microorganisms in this probiotic.

Bifidobacteria are the dominant bacteria in the intestine at birth, but then gradually replaced with Lactobacillus bacteria or bacteria that may adversely affect the living body, the proportion of bifidobacteria in intestinal bacteria gradually decreases It is known to go. Paying attention to this, attempts have been made to utilize bifidobacteria as probiotics. On the other hand, bifidobacteria are obligately anaerobic bacteria, so they are affected by oxygen during growth. However, there is a problem that the viable bacteria rate is not high, and it has been a big problem how to manufacture an oral composition containing a high density of bifidobacteria in a living state at low cost industrially.

  Therefore, several techniques for obtaining an oral composition having a large number of viable bacteria have been proposed. For example, a technique of culturing mixed with Lactococcus lactis (Patent Document 1) A technique of promoting the growth of bifidobacteria using sweet potato and shochu (Patent Document 2) By controlling the timing of sugar addition in the fermentation process A technique for propagating bifidobacteria (patent document 3) A technique for propagating bifidobacteria by dividing the fermentation treatment into two steps (patent document 4) and the like. However, since bacteria other than Bifidobacteria coexist, the use is limited due to special flavors, advanced technology in production management is required, and production cost is required, it is still possible to find satisfactory technology It has not reached.

Japanese Patent Laid-Open No. 10-229819 JP 2009-125055 JP 2012-105577 A JP 2013-090604 A

  It is an object of the present invention to provide a bifidobacteria fermentation method and a growth method for obtaining a composition containing a high concentration of bifidobacteria in a viable state. Accordingly, an object of the present invention is to provide a highly versatile bifidobacteria fermentation method for obtaining a composition containing a high concentration of bifidobacteria in a living bacterium state using an inexpensive raw material without requiring an advanced production technique and The object is to provide a method for proliferating bifidobacteria in the intestine by ingestion, a method for improving flora of intestinal bacteria, and a composition used therefor.

  As a result of intensive studies in view of such circumstances, the present inventors surprisingly fermented pumpkin crushed materials with bifidobacteria, thereby greatly promoting the growth of bifidobacteria and increasing the number of viable bacteria. The bifidobacteria fermented product with a large amount of Bifidobacteria can be obtained, and by simultaneously ingesting bifidobacteria and pumpkin crushed material, etc., the growth of bifidobacteria present in the intestine is promoted, It was greatly increased and the intestinal flora was found to be improved, and the present invention was completed.

That is, this invention provides the growth method of a bifidobacterium of claim | item 1 thru | or 6, the growth promoter, or a fermented product.
Item 1. A method for growing bifidobacteria using a composition containing a processed pumpkin of the following (A) or (B).
(A) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition A bifidobacteria growth promoter comprising a composition comprising the following processed pumpkin (A) or (B).
(A) Pumpkin crushed product of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition Item 3. Pumpkin processed product of the following (C) or (D) Bifidobacteria fermented product containing the composition.
(C) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (D) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition. A method for growing intestinal bifidobacteria using an oral composition containing the processed pumpkin (B) or bifidobacteria of (A) or (B) below.
(A) Pumpkin crushed product of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition An intestinal bifidobacteria growth promoter comprising an oral composition containing the following processed pumpkin (A) or (B) and bifidobacteria.
(A) Pumpkin crushed product of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition Item 6. Pumpkin processed product and bifido of (A) or (B) Item 6. The intestinal bifidobacteria growth promoter according to Item 5, which is an oral composition consisting of only bacteria.

  According to the present invention, since the bifidobacteria are proliferated using a pumpkin that is rich in various nutrients and has a high nutritional value as a food material, the obtained bifidobacteria fermentation product is a bifidobacteria fermentation product of pumpkin. I can say that. Therefore, the bifidobacteria fermented product obtained by the method of the present invention can be used as it is as a resource for high-concentration bifidobacteria in a living state or can be blended in other oral compositions. In addition, since the unique flavor of the pumpkin can be improved by the fermentation treatment of Bifidobacterium, it can also be provided as a pumpkin food material that can be adapted to a wide range of uses. In addition, ingesting pumpkin and viable Bifidobacterium at the same time can increase Bifidobacteria in the intestines and increase the ratio of Bifidobacteria in the intestinal bacteria. A method for bringing the flora closer to the desired state and an oral composition for use in the method can be provided.

The bifidobacteria used in the present invention refers to a bacterium belonging to the genus Bifidobacterium which is a Gram-positive sporeless bacterium. Specifically, Bifidobacterium longum , Bifidobacterium breve , Bifidobacterium bifidum , Bifidobacterium infantis , Bifidobacterium Katenuratamu (Bifidobakuterium catenulatum), Bifidobacterium address Sen Tisdale (Bifidobacterium adolescentis), Bifidobacterium shoe Delon gum (Bifidobacterium pseudolongum), Bifidobacterium animalis (Bifidobacterium animalis), and the like It is done. Among them, Bifidobacterium longum , Bifidobacterium breve , Bifidobacterium bifidum , Bifidobacterium infantis , Bifido Bifidobacterium adolescentis is preferred, Bifidobacterium longum , Bifidobacterium breve , Bifidobacterium infantis is more preferred, Bifidobacterium infantis Most preferred is Bifidobacterium longum . These Bifidobacterium spp. Can be used alone or in combination of a plurality of bacteria.

The pumpkin (nanban) used in the present invention means a fruit of an annual or perennial plant of the genus Cucurbita (scientific name: Cucurbita ). Also known as Nankin, Tounas, Nanjing, Nanban and Bowbra. Pumpkin varieties include Japanese pumpkin (also known as Oriental pumpkin: scientific name Cucurbita moschata Duchesne ex Poir. ), Western pumpkin (scientific name Cucurbita maxima Duchesne ex Lam. ), Pepo pumpkin (scientific name Cucurbita pepo L. ), Mikusta (also known as interspecific hybrids). : Scientific name Cucurbita mixta ), of which Japanese pumpkin and western pumpkin are preferred, and western pumpkin is most preferred. The names of the varieties are: black skinned Nanko, Kogiku Nanto, Sangemon Nanto, Aizu Nanto, Kikuza Nanto, Shikagaya Nanto, Chiba Nanto, Sakura Nanto, Ebisu Nanto, Miyako Nanto, Minehira Nanto, Kuriyutaka, Uchigi Red Skin Chinan-an, Bo-chan Nan-an, Sukuto-nan-an, Fragrance Blue Skin Chin-an, Shintosa Nan-an, Kai-ri Nan-an, Peaceful Goodwill Nan-an, Turban, Delicious, Happard, Zucchini, Hana Zucchini, Mini Zucchini, Gold Thread, Collinky, Butternut Squash, Akon squash, summer squash, spaghetti squash and the like. In the present invention, “pulverized pumpkin” obtained by crushing pumpkins (fruits) can be used, and “pumpkin juice” obtained by squeezing pumpkins and crushed pumpkins can also be used optimally. Can do. The “pumpkin crushed material” is a product obtained by crushing a pumpkin or a pumpkin crushed material to form a puree or paste, and further includes a concentrated product or a dried product obtained by performing a concentration treatment or a drying treatment. The pumpkin crushed material is, for example, heat treated (called blanching) before crushing the pumpkin fruit as necessary, then hammer crusher, electric mill, comitroll, mascolloider, food processor, pulper finisher It can be obtained by crushing using a crusher such as The preferable pumpkin crushed material can use the thing whose insoluble solid content amount is 30 mass% or more with respect to the crushed material whole quantity, and a 40 mass% or more thing can use it preferably. The insoluble solid content mentioned here is an insoluble solid content analysis method for fruit juice beverages, that is, 10 g of a sample is collected in a centrifuge tube and centrifuged at 3000 rpm for 10 minutes using a centrifuge with a rotor radius of 14.5 cm. This is a numerical value obtained by measuring the mass of the supernatant part and the precipitation part separately and measuring the mass percentage of the precipitation part. If the fluidity is poor and difficult to separate under the above-mentioned conditions of centrifugation, the sample that has been diluted with purified water exactly twice and uniformly dispersed is centrifuged, and the mass% of the precipitate is doubled to obtain an insoluble solid content. It is also possible to measure by this. The “pumpkin juice” means that the pumpkin is subjected to a heat treatment (called blanching) as needed, and then pressed as it is, or the crushed material is separated into solid and liquid using a continuous centrifuge or a filter. After removing some or all of the solid content of the pumpkin crushed material, the composition was made liquid, or after adding a suitable amount of purified water to the crushed material and heating it as necessary, it was stirred and solid-liquid separated. The squeezed juice obtained by processing, a squeezed concentrate obtained by concentrating the squeezed liquid, and a dried squeezed product having a shape such as dried powder are included. “Pumpkin crushed material and the like” described below includes both “pumpkin crushed material” and “pumpkin juice” defined above.

  Fermentation using bifidobacteria can be performed by a method exemplified below. The “fermented product” means a fermented product at the time of inoculation of bifidobacteria during the growth of bifidobacteria, that is, during the bifidobacteria fermentation treatment (when a starter is used).

  Before performing fermentation using bifidobacteria, purified water can be appropriately added to the pumpkin crushed material and the like as necessary. For example, when using a dried product or a crushed pumpkin that has been subjected to a concentration treatment, purified water is added and the dried product or a pumpkin that has been subjected to a concentration treatment in order to obtain a state similar to that of the crushed pumpkin before concentration / drying processing It can dissolve and disperse crushed materials. In particular, in pumpkin crushed materials, etc., if the viscosity is too high from the viewpoint of fermentation treatment, or if there is a need to reduce the viscosity of the fermented material for structural reasons of the fermentation treatment device, before the fermentation treatment Add liquid components that do not impair the effects of the present invention, such as purified water, vegetable juice other than pumpkin, fruit juice, etc., to pumpkin crushed materials, etc., and adjust the viscosity to the appropriate value for fermentation treatment, then use bifidobacteria It can be used for fermentation. In addition, when adding other components to pumpkin crushed materials, etc., the amount to be added is not specified, but it is less than the mass of pumpkin crushed materials, etc. when converted to pumpkin crushed materials that have not been concentrated or dried. It is preferable to set the addition amount because the desired effect of the present invention can be easily obtained. That is, it is preferable that 50% by mass or more of the fermented product to be fermented with bifidobacteria is a pumpkin crushed material that has not been subjected to concentration / drying treatment. Furthermore, 75% by mass or more is preferably pumpkin crushed material that has not been subjected to concentration / drying treatment, and most preferably subjected to fermentation using bifidobacteria without mixing other components. In addition, a composition obtained by mixing milk juice, whey, cruciferous plants, fruit juice, etc., with pumpkin crushed materials, etc., which is conventionally used as a fermented product of bifidobacteria, is subjected to fermentation treatment using bifidobacteria. It can also be used as a fermentation product. However, in this case, in the case of a pumpkin crushed product that has not been subjected to treatment such as concentration, the content of the pumpkin crushed product with respect to the total amount of the fermented product is preferably 30% by mass or more, and more preferably 40% by weight or more. More preferably, it is most preferably 50% by mass or more. When using a concentrated or dried pumpkin crushed product obtained by concentrating and drying pumpkin crushed material, the content when converted to the mass of the pumpkin crushed material before each treatment is specified above. It is made to become content to the to-be-fermented product whole quantity to do. Moreover, in the case of pumpkin juice that has not been subjected to processing such as concentration, the content of the juice with respect to the total amount of the fermentation product is preferably 12% by mass or more, more preferably 16% by weight or more, and 20 Most preferably, it is at least mass%. When using a pumpkin juice concentrate or dried product obtained by concentration and drying of pumpkin juice, the content when converted to the mass of the pumpkin juice before each treatment is specified above. It is made to become content to the to-be-fermented product whole quantity to do.

Fermentation using bifidobacteria starts by inoculating the starter of bifidobacteria on the pumpkin crushed material or the like. The bifidus starter can be prepared using conventional methods, and there are no particular restrictions on the properties and composition of the starter. The form of the starter of bifidobacteria used in the present invention may be liquid, frozen or powder. However, when using a starter other than liquid, it is preferable to inoculate after using purified water or a fermented product of starter before inoculation. In addition, when preparing a starter using the pumpkin crushed material etc. which do not add components other than a pumpkin, in order to perform uniform fermentation, water can be previously added to a pumpkin crushed material etc., and it can also adjust to an appropriate viscosity. However, when water is added, the insoluble solid content must be 5% by mass or more. The Bifidobacterium starter which can be preferably used should contain at least 10 8 or more, more preferably 10 9 or more Bifidobacteria per gram of starter at the time of inoculation.
The addition amount at the time of inoculation of the starter is not particularly limited, but usually, it becomes at least 10 6 or more, preferably 10 7 7 viable Bifidobacteria per gram of the fermented product. The amount of starter. The temperature of the pumpkin crushed material at the time of inoculation (when adding the starter to the fermented product) is not particularly limited as long as it is in the range of 30 to 43 ° C, but preferably 35 to 42 ° C, and 36 to 40 ° C. Is most preferred. The temperature of the crushed pumpkin etc. at the time of fermentation shall be 18-45 degreeC. Among these, fermentation is preferably performed in a temperature range of 25 to 42 ° C, and a temperature range of 35 to 42 ° C is most preferable. The fermentation treatment time using the bifidobacteria is appropriately adjusted depending on the temperature conditions at the time of fermentation, etc., but is usually 6 to 30 hours, preferably 8 to 26 hours.

  At the time of the fermentation treatment, a small amount of a plant extract typified by green tea catechin can be added to the Bifidobacterium-fermented product for the purpose of suppressing the growth of bacteria other than Bifidobacterium. These addition amounts are 0.01-0.2 mass% in green tea catechin, for example. Moreover, in order to further improve the growth of bifidobacteria, sugars such as glucose can be added. The addition amount of saccharides is 0.1-10 mass% with respect to a fermented material. Furthermore, a pH adjuster can be added for the purpose of pH control during fermentation of bifidobacteria. Examples of pH adjusters suitable for Bifidobacteria fermentation include sodium acetate, tripotassium phosphate, dipotassium hydrogen phosphate, and potassium dihydrogen phosphate, but are limited to these as long as they have a pH buffering action. It is not a thing. The addition amount of a pH adjuster is 0.01-1 mass% with respect to a fermented material.

  The fermented product obtained by performing the bifidobacteria fermentation treatment of the invention can be used in various forms as long as the viable bacteria rate of the bifidobacteria is not reduced. For example, the fermented product obtained by the method of the present application is solidified and directly or mixed with other solids, and then powdered, granulated, capsule-filled, or tablet / tablet processed to produce an oral composition It can be used as a starter for in-house fermentation such as food and yogurt, and as a raw material for foods and pharmaceuticals. In addition, the fermented product obtained by the method of the present application can be used as an oral composition by blending other raw materials and compounds, or the necessary raw materials for the fermented product can be blended in advance, so that the fermentation treatment of the method of the present application can be performed. What was obtained by performing can also be used as an oral composition as it is. About selection of crushed pumpkin (including selection of remaining degree of solid matter) or juice, viable amount or viability of bifidobacteria after fermentation treatment, processing content after fermentation treatment, fermentation other than pumpkin It is preferable to select as appropriate in consideration of the presence or absence of a substance, the presence or absence of utilization of the physiological function of the insoluble solid content of the pumpkin, and the like.

  For the fermented product used for the fermentation treatment of bifidobacteria of the present invention or the fermented product of bifidobacteria, food raw materials and compounds used in normal oral compositions other than pumpkins are appropriately selected as needed within a range not impairing the effect of the present application. Can be blended.

  Ingredients that can be blended include, for example, fruit juices other than citrus fruits such as apple juice, fruit puree, fruit sauna; juices, extracts, crushed products, pastes of vegetables other than pumpkins; sweeteners (sucrose, glucose, fructose, lactose Sugar alcohols (reduced maltose starch syrup, maltitol, xylitol, sorbitol, erythritol, etc.); oligosaccharides (lactulose, fructooligosaccharides, galactooligosaccharides, isomaltoligosaccharides, raffinose, sucralose, aspartame, acesulfame potassium, stevia extract, etc.) Palatinose oligosaccharides, xylooligosaccharides, etc .; sweeteners (saccharin, saccharin sodium, acesulfame potassium, stevia extract, stevioside, sucralose, neohesperidyl dihydrochalcone, glycyrrhizin, perilartin Saumatine, aspartylphenylalanine methyl ester, methoxycinnamic aldehyde, trehalose, erythritol, sorbitol, palatinose, palatinite, xylitol, maltose, lactitol, fructose, reduced palatinose, glucose, sugar, trithermal sugar, purified honey, unrefined honey, reduced Chickenpox, chickenpox, isomerized sugar (glucose fructose liquid sugar, fructose glucose liquid sugar, etc.); acidulants (fruit acids such as citric acid, gluconic acid, malic acid, tartaric acid, acetic acid, lactic acid, phosphoric acid, succinic acid, glutamic acid Reinforcing agent (L-ascorbic acid, thiamine hydrochloride, riboflavin, pyridoxine hydrochloride, carotene, α-tocopherol, cholecalciferol, calcium lactate, calcium gluconate, calcium carbonate, calcium hydroxide , Magnesium oxide, magnesium hydroxide, magnesium chloride, zinc gluconate, ferrous ammonium citrate, ferric pyrophosphate, etc.); thickening polysaccharides (pectin, carrageenan, guar gum, gum arabic, locust bean gum, alginic acid) Salt, xanthan gum, soybean polysaccharide, gellan gum, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, crystalline cellulose, powdered cellulose, carboxymethylcellulose, ethylcellulose, methylcellulose, etc .; other ingredients (pigment, antioxidant, water-soluble dietary fiber) , Insoluble dietary fiber, animal and plant extracts, amino acids, protein hydrolyzed peptides, trehalose, cyclodextrin, chitosan, hyaluronic acid, phytic acid, yeast extract, vitamins , Minerals, and fragrances).

The present invention can also be used for the purpose of growing bifidobacteria in the intestine, improving the intestinal bacterial flora, and adjusting the condition of the stomach. Specifically, by ingesting the pumpkin juice and / or the composition containing the pumpkin crushed material and bifidobacteria, the bifidobacteria present in the intestines are propagated, or the presence of bifidobacteria in the intestinal bacteria The ratio can be increased and brought closer to or realized in the intestinal bacterial flora that is originally desirable. The pumpkin juice and / or pumpkin crushed material used in the oral composition used in the method for growing Bifidobacteria in the intestine can be greatly improved in the quality retention period of the oral composition because it is a dry product. preferable. When using a pumpkin crushed material in the oral composition, it is 30% by mass or more based on the total amount of the oral composition. When using a pumpkin juice instead of the pumpkin crushed material, it is contained at least 16% by mass based on the total amount of the oral composition. It is preferable. Of course, both may be used in combination. In this case, each blending amount is “Relative value (A) corresponding to the blended amount of pumpkin crushed product when 100% of the pumpkin crushed product is 30% by mass with respect to the total amount of the oral composition”, “Pumpkin juice is When the relative value (B) corresponding to the blended amount of pumpkin juice when “16% by mass” is set to 100 with respect to the total amount of the oral composition, the total value of (A) and (B) exceeds 100. It is preferable to adjust each compounding amount. On the other hand, bifidobacteria need to be present in the oral composition in a viable state. Although not particularly limited, 1 * 10 ⁸ CFU or more exists in the total amount of the oral composition taken at one time. It is preferable to do.

  Hereinafter, the present invention will be specifically described, but the present invention is not limited to the following examples. In the following, “%” means “mass%” unless otherwise specified.

Evaluation of the growth promoting effect of bifidobacteria The growth promoting effect of bifidobacteria was evaluated according to the following method.

(Preparation of evaluation object)
Western pumpkins were cut and pumpkin crushed material was prepared using commitolol. The resulting pumpkin crushed material was subjected to a centrifugal separation process (device name: himac CR21G; manufactured by Hitachi Koki Co., Ltd.); 8000 rpm, room temperature, 10 minutes) to obtain a supernatant. The supernatant was sterilized using a sterilizing filter (0.45 μm) and used as a “pumpkin juice” used for evaluation.

(About the test strain)
The following bifidobacteria were evaluated.
Bifidobacterium longum ( Bifidobacterium longum JCM1217: described as “BL” in Tables 1 and 2)
Bifidobacterium adolescentis JCM1275 (described as “BA” in Table 1)
Bifidobacterium infantis ( Bifidobacterium infantis JCM1222: described as “BI” in Table 1)
Bifidobacterium breve ( Bifidobacterium breve JCM1192: described as “BB” in Table 1)

(About preparation method of inoculated bifidobacteria)
Modified GAM agar “Nissui” (manufactured by Nissui Pharmaceutical Co., Ltd.) was inoculated with each strain, and cultured at 37 ° C. for 24 hours under anaerobic conditions (anaeropack used, manufactured by Mitsubishi Gas Co., Ltd.) One platinum loop of the obtained colonies was collected and inoculated into a modified GAM bouillon “Nissui” (manufactured by Nissui Pharmaceutical Co., Ltd.), and at 37 ° C. for 24 hours under anaerobic conditions (anaeropack use, Mitsubishi Gas) (Manufactured by Co., Ltd.). The bacterial solution after the culture was referred to as “Bifidobacterium culture solution”.

(Evaluation test method)
25 parts by weight of the modified GAM bouillon “Nissui” (manufactured by Nissui Pharmaceutical Co., Ltd.) and 25 parts by weight of the pumpkin juice prepared or sterilized purified water were added and made uniform. 1 to 5% by mass of the inoculated “Bifidobacteria culture solution” is added so that the initial turbidity is in the range of 0.05 to 0.1 (620 nm) with respect to the total amount of the fermented product. The degree (T0, C0: 620 nm) was measured. After the measurement, the cells were cultured under anaerobic conditions (using Anaeropack, manufactured by Mitsubishi Gas Co., Ltd.) at 37 ° C. for 24 hours, and the turbidity (T24, C24: 620 nm) immediately after the cultivation was measured.

The multiplication promotion factor was calculated according to the following formula. The results obtained are shown in Table 1.

Growth promotion factor = ((T24−T0)) / (C24−C0)
T0: Turbidity of test culture solution at the start of culture (620 nm)
T24: Turbidity (620 nm) of the test culture immediately after completion of the culture
T0: Turbidity of control culture solution at the start of culture (620 nm)
C24: Turbidity (620 nm) of the control culture immediately after the end of the culture

  As shown in Table 1, Bifidobacterium longum (27.95), Bifidobacterium adrecentis (3.27), Bifidobacterium infantis (14.23), Bifidobacterium -In breve (15.54), it turned out that a pumpkin crushed material shows the outstanding growth promotion effect. In addition, the degree of the effect varies depending on the species of Bifidobacterium, showing excellent effects in Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium It was found to have the most excellent effect on longum.

Evaluation of the survival effect of bifidobacteria The survival effect of bifidobacteria was evaluated according to the following method.

(Preparation of evaluation object)
As the “pumpkin crushed material”, a western pumpkin was cut and crushed using a comitoroll. As the “broken apple”, apple puree (manufactured by Nagano Kono Co., Ltd.) was used. About both crushed materials, what heated to 90 degreeC and 10 minutes, and returned to normal temperature was used. As a control, Difco MRS Broth (manufactured by Becton Dickinson Company, described as “MRS” in the table) and 0.05% by mass of L-cysteine added to the MRS agar medium (“MRS +” in the table) (Describe)

(About the test strain)
Bifidobacterium longum ( Bifidobacterium longum JCM1217) was used as the Bifidobacterium .

(About preparation method of inoculated bifidobacteria)
“Bifidobacteria culture solution” for inoculation prepared by the method described in the above evaluation was used.

(Evaluation test method)
2% by mass of the “bifidobacteria culture solution” for inoculation prepared by the above method is added to the to-be-evaluated product and the control product prepared above, and the anaerobic condition is maintained at 37 ° C. for 24 hours ( The culture was performed using Anaeropack, manufactured by Mitsubishi Gas Co., Ltd.). After culturing, 1 ml or 1 g was taken and sufficiently stirred using 9 ml of physiological saline until uniform. After stirring, 1 ml was accurately taken and 9 ml of physiological saline was added and stirred sufficiently. This process was repeated and serially diluted. After exactly 100 μl was taken from each serially diluted solution, it was smeared on MRS agar medium (Becton Dickinson) and cultured under anaerobic conditions (anaeropack used, Mitsubishi Gas Co., Ltd.) at 37 ° C. for 48 hours. did. At each serially diluted concentration, the presence or absence of colonies was visually confirmed, and the number of colonies with the highest dilution ratio among those that could be confirmed was counted, and the number of viable Bifidobacteria was determined from the result. The results are shown in Table 2.

As shown in Table 2, the pumpkin crushed material showed a result of 1.5 * 10 ⁹ (CFU / g), and is usually an MRS medium (4.7 * 10 ⁸ (CFU / g) used for culture of bifidobacteria. )) And MRS medium supplemented with cysteine (7.1 * 10 ⁸ (CFU / g)), it was found to have an excellent survival effect on viable bacteria. On the other hand, the viable cell residual effect could not be confirmed in the crushed apple (2.3 * 10 ⁷ (CFU / g)).

Evaluation of intestinal bifidobacterial growth activity by oral ingestion Purchased 5-week-old male mouse C57BL / 6NCrlCrj (SPF) from Japan Charles River Co., Ltd. Groups were grouped on day (10 mice were assigned per group and divided into 4 groups, groups A to D). All groups of mice received the high fat diet D12492 (60 kcal%, Research Diet Inc.) ad libitum. In addition, Bifidobacterium breve powder was given to group B mice, and Bifidobacterium breve powder and pumpkin powder were given to group C mice, respectively. That is, each group was fed as follows.
Group A: High-fat diet feed Group B: High-fat diet feed + Bifidobacterium breve fungus powder Group C: High-fat diet feed + Bifidobacterium breve powder + pumpkin powder
(Pumpkin powder: made by Nippon Powder Chemical Co., Ltd., pumpkin powder)

The dose of Bifidobacterium breve powder is 10 mg (equivalent to 5 × 10 ⁸ CFU), the dose of vegetable powder is 100 mg / 30 g body weight / day, and these are syringes equipped with an oral sonde (disposable syringe, Terumo Co., Ltd.) and orally administered to mice at a frequency of once / day.
The administration period was 84 days, stool was collected on the 84th day, and the ratio of the amount of bifidobacteria to the total amount of bacteria in the stool was consigned to Techno Suruga Lab Co., Ltd. (real-time PCR method) and measured.
Table 3 shows the measurement results of the bifidobacteria growth-promoting effect by ingestion of bifidobacteria and pumpkins using high fat diet-induced obese mice. Specifically, the ratio of the amount of bifidobacteria to the total amount of bacteria in the feces of mice ingested in combination with a high fat diet, bifidobacteria (bifidobacterium breve powder) and pumpkin powder (n = 10 Average value).

  Compared with the control group A (high fat diet group: 0.1), the ratio of bifidobacteria to the total amount of bacteria in the stool of mice was significantly higher in both groups B and C, and group B ( Compared to the bifidobacteria powder alone administration group: 5.0), the “ratio of bifidobacteria to the total amount of bacteria in the stool of mice” in group C (bifidobacteria / pumpkin combination administration group: 6.3) is higher. It was confirmed.

Formulation example: Beverage Fermented product consisting of 30 parts by weight of crushed pumpkin, 20 parts by weight of broccoli puree and flavor, fermented under anaerobic conditions at 37 ° C for 24 hours using Bifidobacterium longum. The fermented bifidobacteria obtained by processing is directly used as a beverage composition.

Claims (6)

A method for growing bifidobacteria using a composition containing a processed pumpkin of the following (A) or (B).
(A) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition A bifidobacteria growth promoter comprising a composition comprising the following processed pumpkin (A) or (B).
(A) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition The bifidobacteria fermented product of the composition containing the pumpkin processed material of the following (C) or (D).
(C) Pumpkin crushed product of 30% by mass or more with respect to the total amount of the composition (D) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition A method for growing intestinal bifidobacteria using an oral composition containing the processed pumpkin (B) or bifidobacteria of (A) or (B) below.
(A) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition An intestinal bifidobacteria growth promoter comprising an oral composition containing the following processed pumpkin (A) or (B) and bifidobacteria.
(A) Pumpkin crushed material of 30% by mass or more with respect to the total amount of the composition (B) Pumpkin juice of 16% by mass or more with respect to the total amount of the composition   6. The intestinal bifidobacteria growth promoter according to claim 5, which is an oral composition comprising only the processed pumpkin of (A) or (B) and bifidobacteria.