Immunostimulating composition, food and beverage including immunostimulating composition, and method for imparting immunostimulating effect to food or beverage

Abstract

To provide: an immunostimulating composition including an insect-derived substance; a food and beverage including the immunostimulating composition; and a method for imparting an immunostimulating effect to a food or beverage.SOLUTION: An immunostimulating composition contains an insect-derived substance. A food and beverage include the immunostimulating composition. There is also provided a method for imparting an immunostimulating effect to a food or beverage. The insect-derived substance is preferably cricket powder and preferably further includes a lactic acid bacterium. The lactic acid bacterium is preferably a lactobacillus strain K-1.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an immunopotentiating composition, foods and beverages containing the immunopotentiating composition, and a method for imparting an immunopotentiating effect to a food or beverage.

Background Art

[0002] Immunity is a system in which immune cells attack antigens such as viruses and bacteria that have invaded the human body, and is an indispensable system for protecting humans from various diseases.

[0003] Such an immune system usually exhibits a certain effect as long as a person lives healthily, but the existence of substances and bacteria with an immunopotentiating effect that improve the action of immunity has also been clarified, and foods and beverages containing them are sold and distributed in various ways.

[0004] Lactic acid bacteria have long been known as having an immunopotentiating effect, and immunopotentiating compositions containing specific strains of lactic acid bacteria and immunopotentiating compositions containing lactic acid bacteria and other compositions have been developed.

[0005] As a prior art document of an immunopotentiating composition containing such lactic acid bacteria, for example, Patent Document 1 contains an acetic acid bacteria culture containing lipopolysaccharide obtained by culturing acetic acid bacteria and heat-killed cells of lactic acid bacteria. The bacterial species of the heat-killed cells of lactic acid bacteria is Enterococcus faecalis, the bacterial species of acetic acid bacteria is Gluconobacter oxydans, and the ratio (m1 / m2) of the mass (m1) of the acetic acid bacteria culture to the mass (m2) of Enterococcus faecalis is 0.0001 to 100, and the acetic acid bacteria culture is cultured in a medium containing soy peptone. An immunopotentiating composition is disclosed.

[0006] By the way, at present, the global food shortage due to population growth has become a problem, and there is a demand for food that is rich in nutrients other than meat and fish and does not require a large amount of cost and energy for production.

[0007] Among such foodstuffs, insects such as crickets and locusts, despite being rich in nutrients such as protein, are attracting attention as a new foodstuff that can replace existing foodstuffs that require costs and energy for breeding, as they are easy to breed.

[0008] In addition, it has been confirmed by research that such insects not only have rich nutrients, but also have various useful effects. For example, in Non-Patent Document 1, antioxidant activity was observed in the ethanol extract of cricket powder, and it was reported that in mice fed the water extract of cricket powder, fatty liver caused by alcohol and permeability in the intestinal tract were suppressed.

[0009] In addition, in Non-Patent Document 2, it has been reported that in humans, the intake of crickets changes the number of intestinal bacteria and reduces the blood TNF-α concentration.

Prior Art Documents

Patent Documents

[0010]

Patent Document 1

Non-Patent Documents

[0011]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0012] However, the immunostimulatory effect of insect-derived substances has not been known until now.

[0013] The present invention has been made in view of such circumstances, and an object thereof is to provide an immunopotentiating composition characterized by containing an insect-derived substance, a food and beverage containing the immunopotentiating composition, and a method for imparting an immunopotentiating effect to a food or beverage.

Means for Solving the Problems

[0014] The inventors conducted intensive studies and found that an insect-derived substance has an immunopotentiating effect. More specifically, it was found that an insect-derived substance has an NK cell activating effect. Furthermore, it was found that when an insect-derived substance and lactic acid bacteria are combined, a more remarkable immunopotentiating effect is exhibited, and the present invention has been completed. That is, the present invention provides the following.

[0015] (1) The first aspect of the present invention is an immunopotentiating composition characterized by containing an insect-derived substance.

[0016] (2) The second aspect of the present invention is the immunopotentiating composition according to (1), wherein the insect-derived substance is a powder of a cricket.

[0017] (3) The third aspect of the present invention is the immunopotentiating composition according to (1), further characterized by containing lactic acid bacteria.

[0018] (4) The fourth aspect of the present invention is the immunopotentiating composition according to (3), wherein the lactic acid bacteria are Lactobacillus sp. K-1 strain.

[0019] (5) The fifth aspect of the present invention is the immunopotentiating composition according to (3), wherein the mixing ratio of the lactic acid bacteria and the insect-derived substance is 1:5 to 2:1.

[0020] (6) The sixth aspect of the present invention is the immunopotentiating composition according to any one of (1) to (5), wherein the immunopotentiating action is an NK cell activating action.

[0021] (7) The seventh aspect of the present invention is a food containing the immunostimulating composition according to any one of (1) to (5).

[0022] (8) The eighth aspect of the present invention is a beverage containing the immunostimulating composition according to any one of (1) to (5).

[0023] (9) The ninth aspect of the present invention is a method for imparting an immunostimulating effect to a food or beverage, which includes adding the immunostimulating composition according to any one of (1) to (5) to the food or beverage.

Advantages of the Invention

[0024] According to the present invention, it is possible to provide an immunostimulating composition characterized by containing an insect-derived substance, a food and a beverage containing the immunostimulating composition, and a method for imparting an immunostimulating effect to a food or beverage.

Brief Description of the Drawings

[0025]

Figure 1

Figure 2

Embodiments for Carrying Out the Invention

[0026] Hereinafter, embodiments for carrying out the present invention (hereinafter simply referred to as "the present embodiment") will be described in detail. The following present embodiment is an exemplification for explaining the present invention, and is not intended to limit the present invention to the following contents. The present invention can be appropriately modified and implemented within the scope of its gist.

[0027] In the present invention, the "immunostimulating composition" means a composition for bringing about an immunostimulating action on animals (humans and non-human animals) or cells derived from animals. In the present invention, the "immunostimulatory effect" includes activation of immunity and the resulting effects of maintaining physical condition and therapeutic effects. The actions included in the activation of immunity include promotion of the production of antibodies (immunoglobulins) such as IgG, IgM, and IgA, and activation of lymphocytes such as B cells, T cells, and NK cells. The immunostimulatory composition of the present invention particularly brings about an activation effect on NK cells.

[0028] <Immunostimulatory composition> The immunostimulatory composition according to one embodiment of the present invention is characterized by containing an insect-derived substance. Hereinafter, the contained components will be described in detail.

[0029] (Insect-derived substance) The insect-derived substance contained in the immunostimulatory composition according to this embodiment is not particularly limited as long as it is a substance derived from an insect that is being bred for food or for which breeding is being considered. However, it is preferably a powder of dried insects. Examples of insects include crickets, locusts, grasshoppers, bees, etc., and among them, crickets are preferred. Examples of types of crickets include crickets that are distributed as food, such as Jamaican crickets, Giant crickets, and European field crickets.

[0030] Also, there is no particular limitation on the form of the above insect-derived substance as long as it can be added to food or beverages. However, when it is a dry powder, the particle size is preferably 30 to 120 μm. By having the particle size within the above numerical range, it becomes easier to mask the prickliness and texture peculiar to insects even when added to food or beverages. Note that the devices and methods for processing insects into dry powder are not particularly limited as long as they are devices used in normal food processing, such as extruders and grinders.

[0031] (Lactic acid bacteria) The immunostimulatory composition according to this embodiment preferably further contains lactic acid bacteria. By containing lactic acid bacteria, a more remarkable immunostimulatory effect can be obtained.

[0032] The lactic acid bacteria are not particularly limited as long as they are lactic acid bacteria with a proven immune activation effect, but among them, lactic acid bacteria K-1 are preferred. Lactic acid bacteria K-1 refers to Lactobacillus casei subsp. casei strain 327 described in WO 2020 / 129888 pamphlet, which is a plant-derived lactic acid bacterium isolated from rice and rice processed products and has antimutagenicity. Note that K-1 is the product name of the lactic acid bacteria of the above strain sold by Kameda Seika Co., Ltd.

[0033] The blending amount of the cells of the lactic acid bacteria contained in the composition for immune activation is generally such that the number of bacteria is around 1.0×10 12 per gram of lactic acid bacteria (it is not necessary to be viable bacteria, but when dead cells are included, it shall be counted as the viable bacteria count before sterilization), and it can be appropriately selected from this amount.

[0034] As described above, the composition for immune activation according to this embodiment can significantly improve the activation effect by containing an insect-derived substance and lactic acid bacteria, compared to administering the insect-derived substance and lactic acid bacteria alone.

[0035] In the composition for immune activation according to this embodiment, the mixing ratio of lactic acid bacteria and the insect-derived substance is preferably 1:5 to 2:1. When the mixing ratio is within the above numerical range, the immune activation effect of the composition for immune activation of the present invention is further improved by a synergistic effect.

[0036] In addition, the composition for immune activation according to this embodiment may, if necessary, add excipients, sweeteners, acidulants, vitamins, minerals, etc. in addition to the insect-derived substance and lactic acid bacteria, and prepare this into forms such as powders, granules, tablets, capsules, paste-like or liquid beverages for use.

[0037] <Food, Beverage> Furthermore, the composition for immune activation according to this embodiment can be incorporated into foods and beverages to impart an immune activation effect. Examples of such foods include, but are not particularly limited to, dairy products such as yogurt and cheese, soups, noodles, processed foods and beverages such as puddings and jams, confectioneries such as chewing gum, candies, tablets, gummy candies, chocolates, biscuits, and snacks, frozen desserts such as ice cream, sherbet, and iced confections, and seasonings such as soy sauce and miso. Examples of beverages include fruit juice drinks, vegetable juices, fruit and vegetable juices, tea drinks, coffee drinks, sports drinks, soft drinks, and health drinks.

[0038] In addition, as a method for imparting an immune activation effect to a food or beverage, not only can it be added during the production of the food or beverage, but the immune activation composition according to the present embodiment may also be in the form of a powder or liquid supplement or seasoning and added when consuming existing foods or beverages.

[0039] As described above, the present invention has been described using embodiments, but it goes without saying that the technical scope of the present invention is not limited to the scope described in the above embodiments. It is obvious to those skilled in the art that various changes or improvements can be made to the above embodiments. Also, it is clear from the description of the claims that forms with such changes or improvements can also be included in the technical scope of the present invention.

Examples

[0040] Hereinafter, the present invention will be described in detail with reference to examples. Note that the present invention is not limited to the examples shown below.

[0041] <Test 1: Evaluation of NK cell activity with cricket powder and Lactobacillus K-1 alone> As a first test, the NK (natural killer) cell activity of cricket powder and Lactobacillus K-1 alone was evaluated. In addition, the concentration involved was also examined. NK cells are one of the main factors of innate immunity, and by evaluating NK cell activity, the immune activation effect of the immune activation composition of the present invention can be evaluated.

[0042] (Preparation of Lactobacillus K-1 suspension) A suspension was prepared using the plant-derived lactic acid bacterium L. casei K-1 (Lacticaseibacillus casei K-1) (Kameda Seika Co., Ltd., 50% of the bacterial cells, Lot 040801). The final exposure amounts of the bacterial cells were adjusted to 30, 10, 3, 1, 0.3, and 0.1 μg / mL, respectively.

[0043] (Preparation of cricket powder suspension) As an insect-derived substance, a suspension was prepared using dried cricket powder (100% cricket) (Ecology Co., Ltd., Lot Z-20220921-CP500). The final exposure amounts were adjusted to 300, 100, 30, 10, 3, and 1 μg / mL, respectively.

[0044] (Cultivation and collection of culture supernatant) Cells suspended in a medium containing 10% FBS and 1% antibiotic solution (hereinafter referred to as the medium) were seeded at 3.0×10 5 cells / well in a 24-well plate and cultured in a CO2 incubator (37°C, 5% CO2) for 6 hours. The medium used was RPMI 1640 with L-glutamine, phenol red, and HEPES (FUJIFILM Wako Pure Chemical Corporation, #189-02145, Lot TPL7017), the FBS was Fetal Bovin Serum (PAN Biotech, #CCP-FBS-BR, Lot P210410), and the cells were a mouse macrophage-like cell line (J774.1). Six hours after seeding, the adjusted L. casei K-1 suspension or cricket powder suspension was added, and the cells were cultured in a CO2 incubator (37°C, 5% CO2) for 24 hours. In this experiment, the suspensions were added to achieve the final exposure amounts shown in Table 1 below.

Table 1

[0045] Thereafter, the culture medium after culturing was collected, centrifuged (4°C, 1,900 rpm, 5 minutes), and the supernatant was collected. Also, to remove impurities, centrifugation was performed again (4°C, 15,000 rpm, 5 minutes), and the supernatant was collected.

[0046] (Preparation of IL-12p40 sample) Next, a sample of IL-12p40, which is a type of interleukin that stimulates NK cells, was prepared. IL-12p40 standards were prepared to be 500, 250, 125, 62.5, 31.3, 15.6, and 7.8 pg / mL, respectively. The measurement samples were prepared by diluting the culture supernatant two-fold with Assay Diluent A and used for measurement.

[0047] (ELISA and calculation of IL-12p40 concentration) Then, ELISA was performed to measure the IL-12p40 concentration. For ELISA, ELISA MAX (registered trademark) Deluxe Set Mouse IL-12p40 (Biolegend, #431604, Lot B355476) was used, and the set included a capture antibody, biotinylated detection antibody, IL-12p40 standard, avidin HRP, TMB substrate reagent, coating buffer, and assay diluent. First, 100 μL / well of the capture antibody was added and incubated at 4°C for 17 hours, and then the wells were washed with wash buffer (PBS + 0.05% Tween-20). Next, assay diluent was added and incubated at room temperature for 1 hour. Thereafter, after washing the wells in the same manner as above, 100 μL / well of the sample and calibration curve solution were added and incubated at room temperature for 2 hours. Thereafter, the wells were washed in the same manner as above.

[0048] Then, 100 μL / well of the detection antibody was added and incubated at room temperature for 1 hour. Thereafter, after washing the wells in the same manner as above, 100 μL / well of the enzyme-labeled antibody (avidin HRP) was added and incubated at room temperature for 30 minutes, and the wells were washed again in the same manner as above. Finally, a color developer was added, and after incubating in the dark at room temperature for 15 minutes, dilute sulfuric acid was added to stop the reaction. Then, the absorbances at 450 nm and 570 nm were measured to calculate Abs450-570nm, and the IL-12p40 concentration in the sample was calculated using the non-linear regression (4-parameter) function of Graphpad PRISM 5.02J from the obtained values.

[0049] (Evaluation) The average value and standard deviation of the IL-12p40 concentration were calculated.

[0050] Figure 1 shows the IL-12p40 concentrations during the treatment with Lactobacillus K-1 and bat powder. Lactobacillus K-1 showed NK cell activation activity at 0.3 μg / mL or higher, and bat powder showed NK cell activation activity at 3.0 μg / mL or higher, and their activities increased in a concentration-dependent manner.

[0051] (Test 2: Evaluation of NK cell activity when bat powder and Lactobacillus K-1 are combined) Next, in the same procedure as in Test 1, the NK cell activity was evaluated when bat powder and Lactobacillus K-1 were combined. The Lactobacillus K-1 suspension was prepared so that the final exposure amount of the cells was 3.0 and 1.0 μg / mL, and the bat powder suspension was prepared so that the final exposure amount was 5.0, 3.0, 2.0, and 1.0 μg / mL. Also, the suspensions added to the medium were added so that the final exposure amounts shown in Table 2 below were obtained.

Table 2

[0052] In Test 2, the significance of the combined use of Lactobacillus K-1 and bat powder was determined by calculating the ratio of the combined treatment to the total value of the single treatment, and when this ratio exceeded 1.3 times, it was judged to have a synergistic effect.

[0053] As a result of Test 2 conducted in this manner, the IL-12p40 concentrations at the time of treatment with Lactobacillus K-1 and cricket powder are shown in Fig. 2. Both Lactobacillus K-1 and cricket powder showed NK cell activation activity at 1.0 μg / mL or higher.

[0054] In addition, Table 3 shows the ratio of the combined treatment to the total value of the single treatments.

Table 3

[0055] As described above, in Lactobacillus K-1 and cricket powder, NK cell activation activity was observed in a concentration-dependent manner. Although there are some differences in the numerical values between Test 1 and Test 2, it is considered that this is due to the difference in the number of cells, and it was determined that there is no problem in the reactivity of the test.

[0056] That is, according to the present invention, it was confirmed that an immunopotentiating composition containing an insect-derived substance can be obtained. It was also confirmed that an immunopotentiating composition containing an insect-derived substance and lactic acid bacteria can be obtained.

Claims

1. An immunopotentiating composition characterized by containing an insect-derived substance.

2. The immunopotentiating composition according to claim 1, wherein the insect-derived substance is a powder of a cricket.

3. The immunopotentiating composition according to claim 1, further characterized by containing lactic acid bacteria.

4. The immunopotentiating composition according to claim 3, wherein the lactic acid bacteria are Lactobacillus K-1 strain.

5. The immunopotentiating composition according to claim 3, wherein the mixing ratio of the lactic acid bacteria and the insect-derived substance is 1:5 to 2:

1.

6. The immunopotentiating composition according to any one of claims 1 to 5, wherein the immunopotentiating effect is an NK cell activation effect.

7. A food containing the immunopotentiating composition according to any one of claims 1 to 5.

8. A beverage containing the immunopotentiating composition according to any one of claims 1 to 5.

9. A method for imparting an immunopotentiating effect to a food or beverage, which includes adding the immunopotentiating composition according to any one of claims 1 to 5 to the food or beverage.

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