Oyster fermentate with enhanced Anti-oxidant, Anti-inflammatory, and Anti-diabetic activities, and uses thereof

Abstract

The present invention relates to an oyster fermentate with enhanced anti-oxidant, anti-inflammatory, and anti-diabetic activities, and uses thereof. The oyster fermentate with enhanced anti-oxidant, anti-inflammatory, and anti-diabetic effects, obtained by inoculating an oyster fermentation medium with Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P), can be utilized in the development of health functional foods and cosmetics.

Description

Oyster fermentation product with enhanced antioxidant, anti-inflammatory and anti-diabetic activities and its use

[0001] The present invention relates to an oyster fermented product having enhanced antioxidant, anti-inflammatory and anti-diabetic activities and its use, and more particularly, to an oyster fermented product having enhanced antioxidant, anti-inflammatory and anti-diabetic activities and its use, obtained by inoculating an oyster fermentation medium with Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) and fermenting it.

[0002]

[0003] Oysters are mollusks belonging to the family Ostreidae of the order Eutaxodonta. There are over 100 species known worldwide, and in the coastal waters of Korea, there are species such as the Pacific oyster (Crassostrea gigas), the rock oyster (Crassostrea nippona), and the striped oyster (Ostrea denselamellosa). Of these, the Pacific oyster is the one that is industrially produced in Korea. Pacific oysters vary in shape, ranging from round to slender and long, but are generally about 10 cm in height and 7 cm in length. In Donguibogam, it is stated that oysters are good for relieving thirst caused by hangovers, and today, various nutritional values ​​of oysters are also reported. Oysters are mainly composed of protein and carbohydrates, and are known to have a high content of taurine (1,006 mg%) and glycogen.

[0004] Meanwhile, marine ecosystems account for 95% of the Earth's biota. Their unique metabolic processes and unique environments, unmatched by terrestrial organisms, offer significant potential for the exploration of diverse new bioactive compounds. While extensive research has already been conducted on terrestrial organisms, research on marine organisms has been limited due to factors such as the lack of traditional marine remedies and the difficulty of collecting marine organisms. Consequently, there is high hope for the future development of unknown natural substances using marine organisms.

[0005] The inventors of the present invention prepared an oyster fermentation medium using oysters with various efficacies as described above, and cultured useful microorganisms in the oyster fermentation medium to complete an oyster fermentation product with enhanced antioxidant, anti-inflammatory, and anti-diabetic activities.

[0006]

[0007] The purpose of the present invention is to provide a fermented oyster product characterized by enhanced antioxidant, anti-inflammatory and anti-diabetic activities through fermentation of useful microorganisms.

[0008] In addition, it is an object of the present invention to provide a method for producing fermented oysters using useful microorganisms.

[0009] In addition, the purpose of the present invention is to provide a health functional food composition for antioxidant and anti-inflammatory purposes containing fermented oyster as an active ingredient.

[0010] In addition, the purpose of the present invention is to provide an anti-diabetic health functional food composition containing oyster fermentation as an active ingredient.

[0011]

[0012] The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0013]

[0014] In order to solve the above problem, the present invention provides a fermented oyster product characterized in that the antioxidant, anti-inflammatory and anti-diabetic activities are enhanced by the fermentation of useful microorganisms.

[0015] The above useful microorganism is characterized in that it is one species selected from Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

[0016] The above oyster is characterized as being a Pacific oyster (Crassostrea gigas).

[0017]

[0018] In addition, the present invention provides a method for producing an oyster fermentation product, characterized by including a step of culturing useful microorganisms in an oyster fermentation medium to obtain an oyster fermentation product.

[0019] The above cultivation is characterized by inoculating useful microorganisms into an oyster fermentation medium and culturing it with shaking at 170 to 190 rpm for 3 to 5 days at 36 to 38°C.

[0020] The above useful microorganism is characterized in that it is one species selected from the group consisting of Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

[0021]

[0022] In addition, the present invention provides an antioxidant and anti-inflammatory health functional food composition comprising, as an active ingredient, an oyster fermented product having enhanced antioxidant, anti-inflammatory, and anti-diabetic activities through fermentation of useful microorganisms.

[0023]

[0024] In addition, the present invention provides an anti-diabetic health functional food composition comprising, as an active ingredient, an oyster fermented product having enhanced antioxidant, anti-inflammatory, and anti-diabetic activities through fermentation of useful microorganisms.

[0025]

[0026] By means of solving the above problem, the present invention provides an oyster fermented product having enhanced antioxidant, anti-inflammatory and anti-diabetic effects by inoculating Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) into an oyster fermentation medium and fermenting the same, thereby enabling the product to be utilized in the development of health functional foods and cosmetics.

[0027]

[0028] The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description of the claims.

[0029]

[0030] FIG. 1 is a graph showing the antioxidant activity according to ABTS radical scavenging ability of oyster fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) according to an embodiment of the present invention.

[0031] FIG. 2 is a graph showing the α-glucosidase inhibitory activity of oyster fermentation product fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) according to an embodiment of the present invention.

[0032]

[0033] The terms used in this specification will be briefly explained, and the present invention will be described in detail.

[0034] The terms used in this invention have been selected from widely used, common terms, taking into account their functions. However, these terms may vary depending on the intentions of those skilled in the art, precedents, the emergence of new technologies, etc. Therefore, the terms used in this invention should be defined based on their meaning and the overall content of the invention, rather than simply their names.

[0035] When a part of a specification is said to “include” a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise stated.

[0036] Below, with reference to the attached drawings, embodiments of the present invention are described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0037] Specific details, including the problems to be solved, means of solving them, and the effects of the invention, are included in the embodiments and drawings described below. The advantages and features of the present invention, and methods for achieving them, will become clearer with reference to the embodiments described below in detail, along with the accompanying drawings.

[0038] Hereinafter, the present invention will be described in more detail with reference to the attached drawings.

[0039]

[0040] The present invention provides an oyster fermented product characterized in that antioxidant, anti-inflammatory and anti-diabetic activities are enhanced by fermentation of useful microorganisms.

[0041] It is preferable that the above useful microorganism is one species selected from the group consisting of Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

[0042] It is preferable that the above oysters are Pacific oysters (Crassostrea gigas).

[0043]

[0044] In addition, the present invention provides a method for producing an oyster fermentation product, characterized by including a step of culturing useful microorganisms in an oyster fermentation medium to obtain an oyster fermentation product.

[0045] It is preferable that the above oyster fermentation medium be a medium containing oyster powder, yeast extract, and NaCl.

[0046] The above cultivation is preferably performed by inoculating useful microorganisms into an oyster fermentation medium and shaking at 170 to 190 rpm for 3 to 5 days at 36 to 38°C, but it is more preferable to inoculate useful microorganisms into an oyster fermentation medium and shaking at 180 rpm for 4 days at 37°C.

[0047] It is preferable that the above useful microorganism is one species selected from Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

[0048]

[0049] In addition, the present invention provides an antioxidant and anti-inflammatory health functional food composition comprising, as an active ingredient, an oyster fermented product having enhanced antioxidant, anti-inflammatory, and anti-diabetic activities through fermentation of useful microorganisms.

[0050]

[0051] In addition, the present invention provides an anti-diabetic health functional food composition comprising, as an active ingredient, an oyster fermented product having enhanced antioxidant, anti-inflammatory, and anti-diabetic activities through fermentation of useful microorganisms.

[0052]

[0053] The above composition may be formulated into a conventional health functional food formulation known in the art. For example, it may be manufactured into general forms such as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, infusions, solutions, and extracts, and may be manufactured into any health food form such as dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, jellies, ice creams, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

[0054] For the formulation of the above composition, a food-grade acceptable carrier or additive may be used, and any carrier or additive known in the art to be usable for the preparation of the formulation to be prepared may be used.

[0055] The above additives may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, fruit pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks may be contained. These additive components may be used independently or in combination.

[0056]

[0057] Hereinafter, to aid understanding of the present invention, examples will be given in detail. However, the following examples are intended only to illustrate the scope of the present invention and are not intended to limit its scope. These examples are provided to more fully explain the present invention to those of average skill in the art.

[0058]

[0059] <Example 1> Optimal fermentation strain using oysters

[0060] Special environmental microbial strains of the European Food Development and Safety Standard (qualified presumption of safety, QSP) were characterized for enzymes such as amylase, protease, lipase, and CMCase, and 12 strains that showed two or more activities and could be used as food ingredients were selected. Among these strains, Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) and Lactobacillus curvatus GH-118-24, which showed excellent antioxidant activity in a fermentation medium using oysters, a specialty crop from Sacheon, Gyeongsangnam-do, Korea, were selected. Lactobacillus curvatus GH-118-24 was deposited at the Biological Resource Center of the Korea Research Institute of Bioscience and Biotechnology and assigned “Accession Number KCTC19050P.”

[0061]

[0062] <Example 2> Production of fermented oyster product

[0063] An oyster fermentation medium was prepared by mixing 2% oyster powder, 3% NaCl, and 0.1% yeast extract in purified water and sterilizing in an autoclave at 121℃ for 15 minutes. Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) selected according to Example 1 was pre-cultured in MRS broth or Marine broth, and the pre-cultured Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) was inoculated into the oyster fermentation medium, and the medium was shaken at 180 rpm for 4 days at 37℃ to recover the oyster fermentation product.

[0064]

[0065] <Example 3> Antioxidant activity analysis (ABTS radical scavenging activity)

[0066] 7 mM ABTS and 2.45 mM potassium persulfate were mixed in a 1:1 ratio and left in a dark room for 16 hours to prepare an ABTS solution (OD value: 0.7 ± 0.02). 20 μl of the fermented oyster product prepared according to Example 2 and 180 μl of the prepared ABTS solution were added to a 96-well plate, mixed well, and reacted at room temperature for 2 minutes, and analyzed by measuring the absorbance at 734 nm.

[0067]

[0068] <Example 4> Analysis of anti-inflammatory effect (DCFH-DA assay)

[0069] The anti-inflammatory effect of the oyster fermentation product manufactured as in Example 2 above was measured by the DCFH-DA test method, which is an intracellular ROS (reactive oxygen species) scavenging efficacy test. Raw264.7 cells were cultured in a 96-well plate with DMEM medium (10% Fetal Bovine Serum, 1% Penicillin-Streptomycin) at 37°C for 24 hours in a 5% CO2 incubator, the culture medium was removed, and 80 μl of DMEM medium containing 5 μM CM-H2DCFDA was added, and the cells were incubated in a 5% CO2 incubator. 2, 37℃ Incubate for 30 minutes, and add oyster fermentation product or positive control (ascorbic acid, α-tocopherol, dexamethasone, L-NIL) prepared as in Example 2 above and incubate in 5% CO 2, The reaction was performed in a 37℃ incubator for 30 minutes. Afterwards, 1㎍ / mL of LPS (Lipopolysaccharides from Escherichia coli O55:B5) was added and the reaction was performed in a 5% CO2 incubator at 37℃ for 24 hours. The inflammatory response was analyzed by measuring the fluorescence of DCF at 485 / 535 nm using an EnVision device.

[0070]

[0071] <Example 5> Analysis of antidiabetic effect (α-glucosidase inhibitory activity)

[0072] The antidiabetic effect was analyzed by measuring the α-glucosidase inhibitory activity using the chromogenic method. In a 96-well plate, a phosphate buffer containing 2 g / L bovine serum albumin and 0.2 g / L NaN3 and the fermented oyster product or positive control (acarbose) prepared as in Example 2 were added and reacted for 5 minutes. A p-nitrophenyl-α-D-glucopyranoside substrate solution was added to the reaction mixture containing the phosphate buffer and the fermented oyster product or positive control, and the reaction was performed at 37°C for 5 minutes. The α-glucosidase inhibitory activity was analyzed by measuring the absorbance at 405 nm using a microplate reader.

[0073]

[0074] <Experimental Example 1> Antioxidant activity of fermented oysters

[0075] According to the above Example 2, the oyster fermentation product was manufactured, and the antioxidant activity using ABTS radical scavenging activity was measured according to the above Example 3 at 0, 24, 48, 72, and 96 hours. As shown in Fig. 1, the oyster fermentation product fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) showed an increase in antioxidant activity at 48 hours and steadily increased for 96 hours. At 96 hours, when the antioxidant activity was the highest, it was confirmed that the antioxidant activity was improved by about 50% compared to the control group. In addition, the oyster fermentation product fermented by Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) showed a rapid increase in antioxidant activity at 48 hours and steadily increased for 96 hours. At 96 hours, when the antioxidant activity was the highest, it was confirmed that the antioxidant activity was improved by about 60% compared to the control group.

[0076]

[0077] <Experimental Example 2> Anti-inflammatory effect of fermented oysters

[0078] As in Example 2 above, the anti-inflammatory effect of oyster fermentation by Lactobacillus curvatus GH-118-24 (KCTC19050P) was tested as in Example 4 and analyzed as shown in Table 1. It was confirmed that the ROS scavenging ability of non-fermented oysters or the oyster fermentation product, which were generated by the inflammatory response induced by LPS, increased compared to the negative control group. In particular, it was confirmed that the ROS scavenging ability of the oyster fermentation product increased more than that of non-fermented oysters. This shows that non-fermented oysters also have an anti-inflammatory effect, but the anti-inflammatory effect increases further when oysters are fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P).

[0079]

[0080] Material name Concentration ROS scavenging activity (%) Negative control LPS 1 ㎍ / mL 0 Positive control Ascorbic acid 40 ㎍ / mL 84.19 α-tocopherol 40 ㎍ / mL 71.77 Dexamethasone 4 ㎍ / mL 91.47 L-NIL 10 ㎍ / mL 77.64 GH-118-24 + Oyster GH 118 24 CG 37 50 ㎍ / mL 32.97 Oyster (NaCl x) GH CG 37 50 ㎍ / mL 19.26

[0081]

[0082] <Experimental Example 3> Antidiabetic activity of fermented oyster products

[0083] The anti-diabetic effect of the oyster fermented product manufactured as in Example 2 was confirmed by measuring the inhibitory activity of α-glucosidase as in Example 5. As shown in Fig. 3, it was confirmed that the oyster fermented product fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) had a greater inhibitory activity of α-glucosidase than that of unfermented oysters. In particular, it was confirmed that the oyster fermented product fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) showed very high activity for the inhibition of α-glucosidase.

[0084] This is a result showing that unfermented oysters and oyster fermented products fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) and Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) have antidiabetic activity, and in particular, it can be seen that the antidiabetic effect of the oyster fermented products fermented by Lactobacillus curvatus GH-118-24 (KCTC19050P) and Bacillus amyloliquefaciens JY-48-5 (KCTC19049P) is more enhanced than that of unfermented oysters.

[0085]

[0086] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred implementation examples and are not intended to limit the scope of the present invention. Therefore, the actual scope of the present invention is defined by the appended claims and their equivalents. The scope of the present invention is set forth in the claims below, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

[0087]

[0088] [Accession number]

[0089] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0090] Accession number: KCTC19050P

[0091] Date of acceptance: 202211121

[0092]

[0093]

Claims

1. Oyster fermentation product characterized by enhanced antioxidant, anti-inflammatory and anti-diabetic activities through fermentation of beneficial microorganisms.

2. In paragraph 1, An oyster fermented product, characterized in that the useful microorganism is one species selected from the group consisting of Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

3. In paragraph 1, Oyster fermentation product characterized in that the oysters above are Pacific oysters (Crassostrea gigas).

4. A method for producing an oyster fermented product, characterized by including a step of culturing useful microorganisms in an oyster fermentation medium to obtain an oyster fermented product.

5. In paragraph 4, The above culturing is a method for producing oyster fermentation, characterized in that the culturing comprises inoculating useful microorganisms into an oyster fermentation medium and culturing it at 36 to 38°C for 3 to 5 days with shaking at 170 to 190 rpm.

6. In paragraph 4, A method for producing fermented oysters, characterized in that the useful microorganism is one species selected from Lactobacillus curvatus GH-118-24 (KCTC19050P) or Bacillus amyloliquefaciens JY-48-5 (KCTC19049P).

7. A health functional food composition for antioxidant and anti-inflammatory purposes containing the fermented oyster product of paragraph 1 as an effective ingredient.

8. An anti-diabetic health functional food composition containing the fermented oyster product of paragraph 1 as an effective ingredient.

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