Lipopolysaccharide, method for producing lipopolysaccharide, and lipopolysaccharide-blended product

Abstract

The purpose of the present invention is to identify a novel substance / factor involved in anti-inflammatory activities of beet, and to provide the substance / factor. This will enable development of health care products such as pharmaceutical products, foods, and cosmetics for suppressing chronic inflammation. Provided is a lipopolysaccharide characterized by being obtained from a bacterium that has been deposited under accession number NITE BP-03839, NITE BP-03840, NITE BP-03841, NITE BP-03842, or NITE BP-03843. Also provided is a pharmaceutical product, a pharmaceutical product for veterinary use, a quasi-pharmaceutical product, a cosmetic, a food, a functional food, a feed, a fertilizer, or a bath additive in which the lipopolysaccharide is blended.

Description

Lipopolysaccharide, method for producing lipopolysaccharide, and lipopolysaccharide formulation

[0001] The present invention relates to lipopolysaccharides, methods for producing lipopolysaccharides and lipopolysaccharide formulations, and in particular to lipopolysaccharide (LPS) derived from bacteria that are uniquely symbiotic to beets.

[0002] Beets are considered a highly nutritious and functional vegetable. They are rich in minerals such as potassium, magnesium, and iron. Other benefits include the nitrite contained in beets, which converts to nitric oxide in the body and dilates blood vessels; betalains, which have antioxidant properties that prevent cell aging; and dietary fiber, which is known to regulate the intestinal environment and improve rough skin (Non-Patent Document 1). Beets are known to maintain immune cell function through the polyphenols and vitamin C found in common vegetables. The pigment betacyanin in beets is known to have anti-inflammatory properties and has been reported to reduce the expression of inflammatory cytokines (TNF-α, IL-6) (Non-Patent Document 2). However, it is not known whether beets activate macrophages, a type of innate immune cell, and increase the expression of anti-inflammatory cytokines (IL-10).

[0003] We have found that lipopolysaccharide (LPS) derived from symbiotic bacteria is attached to plants (Non-Patent Document 3), and that this LPS is taken up by the body and activates the human innate immune system (Non-Patent Document 4). Therefore, it is possible that part of the immunomodulatory effect of beets is related to LPS derived from bacteria that are unique to beets (hereinafter referred to as "beet LPS").

[0004] L. Chen et al., “Beetroot as a functional food with huge health benefits: Antioxidant, antitumor, physical function, and chronic metabolomics activity”, Food Sci Nutr., 2021.11, 9(11), p.6406-6420S. Saito et al., “Metabolic engineering of betacyanin in vegetables for anti-inflammatory therapy”, Biotechnol Bioeng, 2023.05, 120(5), p.1357-1365H. Inagawa et al., “Homeostasis as regulated by activated macrophage. II. LPS of plant origin other than wheat flour and their concomitant bacteria”, Chemical and Pharmaceutical Bulletin, 1992.04, 40(4), p.994-997. “Lipopolysaccharide for Disease Prevention Through the Induction of Priming in Macrophages”, Anticancer Res., 2014.08, 34(8), p.4497-4501

[0005] The present invention clarifies and provides novel substances and factors involved in the anti-inflammatory effects of beets, which will enable the development of healthcare products, such as medicines, foods, and cosmetics, that suppress chronic inflammation.

[0006] The present invention includes the following contents.

[0007] [1] Lipopolysaccharide obtained from bacteria having accession numbers NITE BP-03839, NITE BP-03840, NITE BP-03841, NITE BP-03842, or NITE BP-03843. [2] A method for producing lipopolysaccharide, characterized by obtaining lipopolysaccharide from bacteria having accession numbers NITE BP-03839, NITE BP-03840, NITE BP-03841, NITE BP-03842, or NITE BP-03843. [3] A lipopolysaccharide formulation containing the lipopolysaccharide described in [1] above. [4] The lipopolysaccharide formulation described in [1] or [3] above, further containing a substance of less than 0.5 kDa in beets. [5] An edible plant processed product characterized by containing the lipopolysaccharide described in any one of [1] and [3] to [4] above.

[0008] [6] The edible plant processed product according to any one of [1] and [3] to [5] above, further comprising a beetroot substance of less than 0.5 kDa. [7] The edible plant processed product according to [5] or [6] above, characterized in that the edible plant processed product is a beetroot processed product. [8] The edible plant processed product according to any one of [5] to [7] above, characterized in that the edible plant processed product is dried vegetables or dried edible plant powder. [9] A food product comprising the edible plant processed product according to any one of [5] to [8] above.

[10] The lipopolysaccharide formulation according to [3] or [4] above, characterized in that the lipopolysaccharide formulation is a pharmaceutical product, veterinary drug, quasi-drug, cosmetic, food, functional food, feed, fertilizer, or bath additive.

[0009] Beet LPS, derived from bacteria isolated from beets, activates macrophages. Compared to the known Pantoea agglomerans LPS, it exhibits unique macrophage activation, including a higher ability to induce the anti-inflammatory cytokine IL-10. This effect was demonstrated by the action of polymyxin B, which is clearly LPS. Furthermore, it was shown that macrophage activation is not the result of the action of beet LPS alone, but rather an interaction with low-molecular-weight substances in the beet. This may enable the development of healthcare products, such as medicines, foods, and cosmetics, that suppress chronic inflammation.

[0010] In this specification, beets refer to beets listed in the 2020 edition (8th revision) of the Standard Tables of Food Composition in Japan.

[0011] As used herein, "edible plants" refer to plants consumed by humans. As used herein, edible plants are not limited to those consumed by humans, but include vegetables, grains, potatoes, legumes, nuts and seeds, fruits, mushrooms, and algae. Vegetables are preferred, with beets being particularly preferred. Edible plants may be used singly or in any combination of two or more. Furthermore, edible plants may be used as is or after various processes (e.g., drying, heating, removing lye, peeling, removing seeds, ripening, salting, peel processing, etc.). Even for foodstuffs whose edible parts (e.g., edamame soybeans and green peas) are considered vegetables, the entire plant (e.g., soybeans and peas) combined with the inedible parts (e.g., pods) can be used to determine whether they are legumes. Furthermore, edible plant classification can be determined based on the entire plant, including the inedible parts. Specifically, for example, by referring to the classifications listed in the "Standard Tables of Food Composition in Japan, 2015 Edition (7th revision) Supplement 2018" (a food composition table established by the Ministry of Health, Labour and Welfare, see especially Table 1 on page 236), such as vegetables, grains, potatoes, beans, nuts and seeds, fruits, mushrooms, and algae, it is possible to understand what foods fall under the category of edible plants in the present invention.

[0012] The properties of the edible plant processed product of the present invention are not limited, but are preferably one or more selected from edible plant powder, edible plant paste, and aqueous extract of edible plants. For example, a processed product obtained by subjecting an edible plant to heat treatment (e.g., 80°C or higher) such as drying, roasting, or hot water extraction is preferred. The edible plant processed product of the present invention is preferably dried vegetables or dried edible plant powder. Examples of the present invention will be described below, but the present invention is not limited to the following examples.

[0013] [Evaluation of macrophage activation by beet hot water extract] (1) Method 1) Preparation of beet extract

[0014] Beets with the skin on were washed with water and cut into 1cm square cubes with a knife. The cut beets were then heated in a steamer at a temperature of 90°C or higher for 15 minutes. The cooked beets were then spread evenly on a perforated tray, and the tray was placed in a hot air dryer set at 70°C. The beets were dried (7-8 hours) until the moisture content reached 6% or less, creating beet chips. The chips were then crushed in a grinder (Osaka Chemical) and passed through a 355μm mesh to create dried beet powder.

[0015] Approximately 2 g of the dried beet powder prepared above was weighed and transferred to a 50 mL tube. Distilled water was added to a concentration of 100 mg / mL, and the mixture was heated in an autoclave at 90°C for 20 minutes. After cooling, the mixture was stirred 10 times for 30 seconds each in a vortex mixer (Vortex-genie2, Scientific Industries), followed by ultrasonic treatment at 37°C for 15 minutes. After ultrasonic treatment, the mixture was centrifuged and the collected supernatant was used as the beet extract.

[0016] 2) LPS amount measurement method LPS amount was measured using a toxinometer (Fujifilm Wako Pure Chemical Industries) as a limulus value (value obtained by the limulus test, converted value of LPS produced by the control standard endotoxin E. coli UKT-B).

[0017] 3) Preparation of samples for evaluation of macrophage activation by beet extract RAW264.7 cells were subcultured in RPMI1640 medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin. The cells were cultured in a T25 culture flask, with 0.25 × 10 cells / ml added every 3 or 4 days. 5 The cells were subcultured at 1000 cells / mL. They were cultured in a 37°C 5% CO2 incubator (hereinafter referred to as the incubator). All test procedures were performed in a clean bench. Cells precultured in a T25 culture flask were detached from the wall by pipetting, and the resulting cell suspension was transferred to a conical tube. The tube was centrifuged at 1000 rpm for 5 minutes at room temperature, the supernatant was discarded by decantation, and the cells were recovered. After loosening the cells by tapping, culture medium was added and the cells were uniformly suspended by pipetting. These RAW264.7 cells were plated at 5 x 10 5 Cells were seeded at 0.5 mL per well and cultured for 3 hours in a 37°C incubator. Then, 0.5 mL of culture medium containing or not containing 2x concentrations of each test substance was added to the wells. LPS derived from Pantoea agglomerans (LPSp: Funakoshi, mac0001) was used as a positive control. Samples other than the medium were diluted with medium to an LPS concentration of 10 ng / mL. After addition, the cells were cultured in a 37°C 5% CO2 incubator, and the culture medium was collected after 4 hours.

[0018] 4) IL-10 Gene Expression Analysis Method: Total RNA extract (40 μL) was prepared from cells after removing the culture medium using the RNeasy Mini Kit (QIAGEN) according to the manufacturer's instructions. Absorbance and RNA concentration were measured using 10 μL of the RNA extract using NanoVue Plus (GE Healthcare Japan). Two μg of the recovered total RNA was treated with ReverTra Ace® qPCR RT Master Mix with gDNA Remover (Toyobo Co., Ltd.) according to the kit's instructions to degrade chromosomal DNA, followed by reverse transcription to synthesize cDNA. Expression of GAPDH (a housekeeping gene) and IL-10 was analyzed by qPCR.

[0019] (2) Results The results of IL-10 gene expression in RAW264.7 cells for each sample are shown in Table 1. The average in the table represents the arithmetic mean of the IL-10 expression fold for each sample. Medium: Medium only LPSp: Medium with LPSp added Beetroot extract: Medium with beetroot extract added Beetroot extract showed a higher IL-10 expression fold than LPSp.

[0020]

[0021] Polymyxin B (PB), an LPS inhibitor, was added to RAW264.7 cells, and IL-10 gene expression was measured in response to LPSp and beet extract. The PB-induced inhibition rate was calculated by comparing the fold-inhibition of IL-10 expression with that before PB addition. The results are shown in Table 2.

[0022] *PB(-): No PB added. PB(+): PB added. Both samples were shown to be inhibited by more than 90% by the addition of PB, indicating that this activity was due to LPS.

[0023] LPS is a lipopolysaccharide present in the cell membrane of gram-negative bacteria. These results suggest that the substance in beets that has the characteristic macrophage activation effect is LPS derived from gram-negative bacteria that live symbiotically with beets.

[0024] [Search for substances with characteristics of beet hot water extract] Next, we investigated whether beet-derived components other than LPS have any effects using molecular fractionation. (1) Method 1) Preparation of beet extract As described in Example 1.

[0025] 2) Extract Fractionation The beet hot water extract was centrifuged using a 10 kDa centrifugal ultrafiltration filter (Merck Millipore Ltd.) to separate the inner fraction (ultrafiltrate inner fraction) containing LPS and the outer fraction (ultrafiltrate outer fraction) containing LPS. Distilled water was added to the inner fraction, and the centrifugation (washing procedure) was repeated five times. Each fraction, including the distilled water used for washing, was lyophilized, and distilled water was added.

[0026] Furthermore, the ultrafiltrate (less than 10 kDa) was dialyzed against distilled water using a 0.5 kDa dialysis membrane (Funakoshi). Dialysis was performed five times, and the fluid inside the dialysis membrane (0.5-10 kDa) and the fluid outside the dialysis membrane (less than 0.5 kDa) were collected. The fluid outside the dialysis membrane was lyophilized and then reconstituted with distilled water. 3) IL-10 Gene Expression Analysis Additional samples were added and analyzed as described in Example 1.

[0027] (2) Results Each sample was added to RAW264.7 cells, and the results of examining IL-10 gene expression are shown in Table 3. The average in the table represents the arithmetic mean value of the IL-10 expression fold for each sample. LPSp: LPSp added to the medium. Beetroot extract: Beetroot extract added to the medium. Ultrafiltrate inner solution: Ultrafiltrate inner solution added to the medium. Ultrafiltrate outer solution: Ultrafiltrate outer solution added to the medium. Ultrafiltrate inner solution + ultrafiltrate outer solution: Ultrafiltrate inner solution and ultrafiltrate outer solution added to the medium.

[0028]

[0029] These results indicate that IL-10 gene expression in the ultrafiltrate inner solution was significantly lower than that in the beetroot extract. Furthermore, IL-10 gene expression was barely observed in the ultrafiltrate outer solution. The fact that some substance in the ultrafiltrate outer solution is responsible for the characteristics of beetroot extract was confirmed by reconstituting the ultrafiltrate inner solution and the ultrafiltrate outer solution. This indicates that the ultrafiltrate outer solution contains a substance that induces the characteristics of beetroot extract.

[0030] Here, polymyxin B (PB), an LPS inhibitor, was added to RAW264.7 cells, and IL-10 gene expression in the ultrafiltrate was measured, and the results are shown in Table 4. The inhibition rate by PB was calculated in the same manner as in Example 1. LPSp: LPSp was added to the medium. LPSp + ultrafiltrate: LPSp and ultrafiltrate were added to the medium. Beet extract: Beet extract was added to the medium. Ultrafiltrate: Ultrafiltrate was added to the medium. Ultrafiltrate + ultrafiltrate: Ultrafiltrate and ultrafiltrate were added to the medium.

[0031] *PB(-): No PB added. PB(+): PB added. As shown above, the addition of PB inhibited IL-10 expression by more than 90%, indicating that this activity was due to LPS.

[0032] Furthermore, in order to limit the molecular weight of low-molecular-weight substances contained in the 10 kDa ultrafiltrate external solution, a 0.5 kDa dialysis membrane was used to create a dialysis membrane internal solution (0.5-10 kDa) and a dialysis membrane external solution (less than 0.5 kDa), and their effects were examined. The results are shown in Table 5. The average in the table represents the arithmetic mean value of the IL-10 expression fold for each sample. LPSp: LPSp added to the medium. Beetroot extract: Beetroot extract added to the medium. Ultrafiltrate internal solution: Ultrafiltrate internal solution added to the medium. Ultrafiltrate internal solution + dialysis membrane external solution: Ultrafiltrate internal solution and dialysis membrane external solution added to the medium.

[0033]

[0034] The beet extract showed a characteristic of increasing IL-10 expression by reconstituting the ultrafiltrate inner fluid and the dialysis membrane outer fluid. Based on the above, it was narrowed down to substances of less than 0.5 kDa that contain low molecular weight substances that increase IL-10 expression in the ultrafiltrate outer fluid.

[0035] [Isolation of microorganisms with characteristics of beet hot water extract] (1) Method for isolating beet indigenous bacteria

[0036] 1) Preparation of beet agar medium (beet extract 25, 75%, yeast extract 0.5%, amphotericin 5 μg / mL, vancomycin 10 μg / mL, agar 1.5%)

[0037] (1): Hokkaido-grown beets were cut into approximately 1 cm cubes and an amount of water for injection (Otsuka Pharmaceutical Co., Ltd.) equal to the weight of the beets was added. After pulverization using a homogenizer, the mixture was centrifuged (3500 rpm, 10 minutes) and the supernatant was collected (50% beet extract). The 50% beet extract was heated at 68°C for 30 minutes and then freeze-dried. Distilled water was added to the freeze-dried product in an amount equal to the original 50% beet extract concentration (50%) and three times the original concentration (150%). Amphotericin B (Fujifilm Wako Pure Chemical Corporation) was added to a final concentration of 10 μg / mL, and vancomycin (Fujifilm Wako Pure Chemical Corporation) was added to a final concentration of 20 μg / mL.

[0038] (2): In a separate container, agar (powder, Fujifilm Wako Pure Chemical Industries, Ltd.) and yeast extract (Nacalai Tesque, Inc.) were added to distilled water to final concentrations of 3% and 1%, respectively, and autoclaved at 121°C for 20 minutes.

[0039] (3): When the solution (2) reached approximately 60°C, it was mixed with an equal amount of (1) to prepare a final concentration of 25% and 75% beetroot medium. 25 mL of this mixture was dispensed into 10 cm petri dishes.

[0040] 2) Bacterial isolation method The following procedures were performed aseptically in a clean bench. The surface of the beet was scrubbed and washed with Kimwipes soaked in water for injection. The skin of the washed area was collected and cut into small pieces with scissors. The skins of three beets were mixed, and an equal amount of water for injection was added by weight and stirred with a vortex mixer (suspension). This suspension was diluted with physiological saline to 5 x 10 3 Double and 5x10 4 The diluted suspension was diluted 1:100. 100 μL of the diluted suspension was plated on beet agar medium (beet extract 25%, 75%) and LB agar medium.

[0041] 3) Bacterial culture method The plates spread on each medium were cultured under aerobic and anaerobic conditions. 4 The plates were cultured at 25°C for 3 to 5 days. 3 The plates containing the diluted cultures were placed in an anaerobic pouch (Anelopouch Kenki, manufactured by Sugiyamagen Co., Ltd.) and cultured for 3 days in an incubator set at 25°C. Two hours after the start of culture, the anaerobic indicator was confirmed to show an oxygen-free state (0.1% or less).

[0042] (2) Results Aerobic culture was carried out for 5 days and anaerobic culture for 3 days on beet extract medium (25, 75%). The number of colonies obtained is shown in Table 6. Beet extract 25% medium: beet extract medium with a concentration of 25% Beet extract 75% medium: beet extract medium with a concentration of 75%

[0043] Colonies from each plate were scraped off with a cell scraper, and water for injection was added to a wet cell weight of 10-100 mg / mL, followed by heating at 90°C for 20 minutes (hot water extract of cultured bacterial populations). The LPS content of the hot water extract of each cultured bacterial population is shown in Table 6. Each cultured bacterial population contained LPS that could be measured with Limulus enzyme.

[0044]

[0045] Next, IL-10 gene expression was examined using hot water extracts of the cultured bacterial population with and without PB to confirm that this activity was derived from LPS. As shown in Table 7, the IL-10 expression fold was significantly higher in anaerobic culture in 25% beet extract medium than in 75% aerobic and anaerobic cultures, relative to LPSp (p<0.05 in the t-test). Furthermore, the inhibition rate by PB exceeded 90% in aerobic and anaerobic cultures in 25% beet extract medium, and in aerobic cultures in 75% beet extract medium. Based on these findings, the anaerobic culture in 25% beet extract medium (number of colonies: 6) and the aerobic culture in 75% beet extract medium (number of colonies: 13) were selected. The inhibition rate by PB was calculated using the same method as in Example 1.

[0046] *PB(-): No PB added. PB(+): PB added. *: Significant P<0.05 in t-test.

[0047] [Identification of microorganisms with characteristics of beet hot water extract] Beets were cultured anaerobically in a 25% beet extract medium and aerobicly in a 75% beet extract medium. From the resulting bacterial population, strains were selected, and the genus was identified using the 16S rRNA method. The strains were then deposited. (1) Identification method 1) Preparation method of LB agar medium

[0048] Agar was added to LB medium to a concentration of 1.5%, and the mixture was autoclaved at 121°C for 20 minutes. After cooling to approximately 60°C, reagents were added to give final concentrations of amphotericin B of 5 μg / mL and vancomycin of 10 μg / mL, and 25 mL of each was dispensed into petri dishes.

[0049] 2) Identification of the bacteria: Nineteen colonies selected from replicas (stored at 4°C) of 25% beetroot medium (anaerobic culture) and 75% beetroot medium (aerobic culture) were streaked onto new LB medium. After aerobically culturing at 25°C for two days, single colonies were streaked onto new medium. After culturing at 25°C for two days and confirming the colonies, the plates were sent to Techno Suruga Lab, a subcontractor, to obtain partial 16s rDNA sequence analysis data.

[0050] 3) Hot phenol water extraction Hot phenol water extraction was performed using the method of Westphal et al. Water for injection was added to the wet cells to a concentration of 100 mg / mL and stirred using a vortex mixer. An equal volume of 90% phenol to the suspension was added and mixed using a vortex mixer. The mixture was heated in a water bath set at 68°C for 20 minutes. During heating, the mixture was stirred for approximately 10 seconds every 5 minutes. After cooling to room temperature, the mixture was centrifuged (3500 rpm, 20 minutes, room temperature) and the aqueous layer was recovered. An equal volume of water for injection to the recovered aqueous layer was added, and phenol extraction was performed again.

[0051] The collected aqueous layer was diluted 10-fold with water for injection. To remove phenol from this solution, ultrafiltration was performed using a centrifugal 10 kDa ultrafiltration filter, and the LPS content in this solution was measured by the Limulus method.

[0052] 4) IL-10 Gene Expression Analysis Method IL-10 gene expression analysis was performed for each sample as described in Example 1. Ultrafiltrate was used as the low molecular weight substance here, but as mentioned above, any low molecular weight substance that actually increases the IL-10 expression fold is a substance of less than 0.5 kDa.

[0053] (2) Results The partial 16s rDNA sequences of the 19 strains obtained above were determined by Techno Suruga Lab, and the sequence data was subjected to a BLAST search against the international base sequence database to identify the genus of the bacteria. Of these, three strains from the 25% beet medium (anaerobic culture) and two strains from the 75% beet medium (aerobic culture) were deposited at the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation. Table 8 shows the genus and accession number of each strain.

[0054]

[0055] The IL-10 gene expression of the deposited bacterial cells was measured, and the fold increase in the expression level when the low molecular weight substance was added to the sample alone compared to before the addition of the low molecular weight substance was calculated. The results are shown in Tables 9 and 10.

[0056]

[0057]

[0058] These results revealed that the IL-10 expression fold increase in the sample alone was higher than that in LPSp (LPS derived from BR-4, BR-6, BR-15, and BR-19 strains), or that the IL-10 expression fold increase was due to the addition of low molecular weight substances (ultrafiltrate of less than 10 kDa was used, but in reality substances of less than 0.5 kDa were effective), with the increase being more than 20-fold compared to the sample alone (LPS derived from BR-1 and BR-15 strains) in LPS derived from beet isolates.

[0059] These findings demonstrate that beets have anti-inflammatory properties due to the interaction between LPS derived from bacteria specific to beets and low molecular weight substances that modify the action of LPS. New substances related to the immunoregulatory function of beets may enable the development of pharmaceuticals, veterinary medicines, quasi-drugs, cosmetics, food, functional foods, feed, fertilizers, and bath additives to suppress chronic inflammation, as well as healthcare products such as foods that incorporate edible plant products obtained by processing edible plants such as vegetables, fruits, and grains, particularly root vegetables.

[0060] All publications cited herein are incorporated herein by reference in their entirety.

[0061] Name of depository institution: National Institute of Technology and Evaluation, Patent Microorganisms Depositary Address of depository institution: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan Date of deposit: March 2, 2023 Accession number: (1) NITE BP-03839 (2) NITE BP-03840 (3) NITE BP-03841 (4) NITE BP-03842 (5) NITE BP-03843

Claims

1. Lipopolysaccharide obtained from a bacterium having accession number NITE BP-03839, NITE BP-03840, NITE BP-03841, NITE BP-03842, or NITE BP-03843.

2. A method for producing lipopolysaccharide, which comprises obtaining lipopolysaccharide from a bacterium having accession number NITE BP-03839, NITE BP-03840, NITE BP-03841, NITE BP-03842, or NITE BP-03843.

3. A lipopolysaccharide preparation comprising the lipopolysaccharide according to claim 1.

4. A lipopolysaccharide preparation according to claim 1 or claim 3, further comprising a substance less than 0.5 kDa in beets.

5. An edible plant processed product, characterized in that the lipopolysaccharide according to any one of claims 1 and 3 to 4 is blended therein.

6. An edible plant processed product according to any one of claims 1 and 3 to 5, further comprising a substance of less than 0.5 kDa contained in beets.

7. The edible plant processed product according to claim 5 or 6, characterized in that the edible plant processed product is a beetroot processed product.

8. The edible plant processed product according to any one of claims 5 to 7, characterized in that the edible plant processed product is a dried vegetable or a dried edible plant powder.

9. A food comprising the edible plant processed product according to any one of claims 5 to 8.

10. The lipopolysaccharide formulation according to claim 3 or 4, characterized in that the lipopolysaccharide formulation is a pharmaceutical product, an animal drug, a quasi-drug product, a cosmetic product, a food product, a functional food product, an animal feed, a fertilizer, or a bath additive.

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