Method for preparing mushroom melanin solution and mushroom melanin solution prepared thereby

Abstract

The present invention relates to a method for preparing mushroom melanin by using extracted mushroom juice made by grinding selected mushrooms to a predetermined size, and then reacting, by injecting, a substrate and an enzyme group having an enzyme, particularly tyrosinase, as a main component, said group being included in the mushroom juice, and then recovering the mushroom melanin so as to apply the mushroom melanin to various food materials and additives.

Description

Method for producing mushroom melanin solution and mushroom melanin solution produced thereby

[0001] The present invention relates to a method for preparing a mushroom melanin solution and a mushroom melanin solution prepared thereby. The present invention also relates to applications of the mushroom melanin solution.

[0002] Melanin is a general term for black or brown pigments that exist in tissues such as the skin, eyes, fur, and hair of various animals. Representative enzymes that play a major role in the production of melanin include tyrosinase, laccase, and mono- or deoxy-oxidase, and melanin can be synthesized using the above enzymes as catalysts and amino acids such as tyrosine, cysteine, histidine, and lysine or their derivatives as substrates, or hydroxyphenyl, pyruvic acid, homogentisate, dihydroxynaphthalene, and polyphenolic acid. More specifically, tyrosine can be converted into various forms of melanin by tyrosinase, an oxidizing enzyme that exists in animals, plants, microorganisms, and fungi in nature, and the above enzyme groups.

[0003] Tyrosinase can convert tyrosine into L-dopa as a reaction intermediate, and oxygen is used in this process, and various intermediates (e.g., dopaquinone, dopachrome, hydroxyphenyl pyruvate, homogentisate, etc.) are then produced. These intermediates are ultimately used to synthesize various colored melanins (e.g., eumelanin, pheomelanin, pyomelanin, and allomelanin).

[0004] The present invention relates to a method for producing a mushroom melanin solution using various enzymes, including tyrosinase, from mushroom powder or mushroom juice obtained by crushing mushrooms and then squeezing the juice, and relates to a mushroom melanin solution obtained through a natural enzyme oxidation reaction from mushroom juice extracted from various types of mushrooms, and a mushroom melanin solution usable as a food additive.

[0005] [Prior Art Literature]

[0006] [Patent Document]

[0007] (Patent Document 1) Korean Patent Publication No. 10-2014-0119502 (October 10, 2014)

[0008] (Patent Document 2) Korean Patent Publication No. 10-2010-0049794 (May 13, 2010)

[0009] The present invention provides a method for producing mushroom melanin by using mushroom juice made by grinding mushrooms into a certain size and utilizing enzymes contained therein, specifically, enzymes having tyrosinase as a main component, and applying the same to various food materials (e.g., food additives).

[0010] The present invention specifically provides the following.

[0011] (1) (a) A step of crushing mushrooms to obtain mushroom powder, or crushing mushrooms, mixing them with water, and then squeezing the juice with a cotton cloth or juicer to obtain mushroom juice, and

[0012] (b) a step of reacting the mushroom powder or mushroom juice with a substrate to obtain a mushroom melanin solution,

[0013] A method for producing a mushroom melanin solution, wherein the substrate is selected from the group consisting of amino acids selected from L-tyrosine, L-cysteine, L-histidine or L-lysine; polyphenols selected from phloretin, resveratrol, P-coumaric acid, caffeic acid, isoflavonoids, flavonoids, catechol, catechin, tyrosol, tannic acid or gallic acid; and an extract of a plant containing polyphenols selected from blueberry extract, tea leaf extract, soybean extract or a mixed extract thereof.

[0014] (2) A method for producing a mushroom melanin solution according to (1), wherein the mushroom is a pine mushroom, a pine mushroom, a oyster mushroom or a oyster mushroom.

[0015] (3) A method for producing a mushroom melanin solution, further comprising, after step (b), (c) a step of low-temperature sterilization at 60 to 65°C for 25 to 35 minutes, high-temperature sterilization at 120°C or higher for 4 minutes or more, or ultraviolet (UV) sterilization for 30 minutes or more.

[0016] (4) In (1), in step (b), a method for producing a mushroom melanin solution wherein the substrate is 4 to 8 mM L-tyrosine.

[0017] (5) In step (1), a method for producing a mushroom melanin solution, wherein the L-tyrosine is contained in a food.

[0018] (6) A method for producing a mushroom melanin solution, comprising, in (3), an additional step of concentrating by centrifugation at 6,000 to 12,000 xg (rcf) for 20 to 35 minutes after step (c).

[0019] (7) A mushroom melanin solution manufactured according to any one of the manufacturing methods of (1) to (6).

[0020] (8) In (7), the mushroom melanin solution is a mushroom melanin solution containing melanin or eumelanin.

[0021] (9) In (7), the mushroom melanin solution further comprises a thickener selected from the group consisting of gellan gum, xanthan gum, carboxymethylcellulose, konjac gum, tamarind gum, guar gum, alginic acid, carrageenan, and sodium caseinate.

[0022] (10) In (7), the mushroom melanin solution further comprises an emulsifier selected from the group consisting of lecithin, saponin, cocobetaine, polysorbate, glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, polyoxyethylene fatty acid ester, and propylene glycol fatty acid ester.

[0023] (11) (7), the mushroom melanin solution is prepared in the form of a concentrate, paste, freeze-dried, powder or granule formulation.

[0024] (12) A food additive containing a mushroom melanin solution of (11).

[0025] (13) In (12), the food additive is a food additive in the form of a sauce.

[0026] (14) (13), the food additive is a food additive for use in a food selected from the group consisting of bakery, rice cake, tofu, noodles, red pepper paste, soybean paste, soybean paste, pasta sauce, and food dressing.

[0027] The present invention optimizes the mushroom grinding process to maximize the production of tyrosine oxidase complexes, primarily tyrosinase, from mushroom juice extracted from ground mushrooms. Furthermore, by providing various substrates, the efficiency of mushroom melanin production can be increased, and the desired melanin color and quality can be controlled. This can improve color, flavor, and functionality in the food industry.

[0028] The present invention differs from existing animal-derived melanin production methods (e.g., squid ink) in that it can increase the efficiency of mushroom melanin production. Specifically, compared to existing animal-derived melanin, the present invention has the advantage of being marketed as a vegan food product because it is a plant-based raw material derived from mushrooms. Furthermore, it is environmentally friendly compared to animal-derived melanin, which is affected by the surrounding environment.

[0029] Furthermore, the present invention has the advantage of being applicable to foods that meet various dietary and dietary requirements, in addition to vegan foods. For example, the present invention can be utilized in the production of halal and kosher foods.

[0030] The present invention can be manufactured while preserving the flavor and nutrients of mushrooms, and can be manufactured in various formulations such as liquid and powder forms, so that it can be used as a food additive and food.

[0031] The present invention is a technology that utilizes various mushrooms, such as shiitake mushrooms, oyster mushrooms, oyster mushrooms, and oyster mushrooms, and has utility value in that it can be utilized as an edible coloring, including the flavor and nutrients of mushrooms.

[0032] Figure 1 is a process diagram for manufacturing mushroom juice and mushroom melanin solution according to the present invention.

[0033] Figure 2 is a drawing confirming the reactivity between tyrosinase and L-tyrosine extracted according to the type of mushroom.

[0034] Figure 3 is a diagram showing the mushroom melanin production amount according to the control of the concentration of shiitake mushrooms and the concentration of L-tyrosine.

[0035] Figure 4 is a diagram showing the consumption pattern of L-tyrosine, a reaction substrate, over time by measuring the absorbance corresponding to dopachrome.

[0036] Figure 5 shows various formulations of mushroom melanin solution ((a) dry, (b) paste, (c) liquid formulation).

[0037] Figure 6 is an example of how mushroom melanin solutions can be applied as food additives. (a) and (b) are examples of using squid ink as a control food additive, and (c) and (d) are examples of using the mushroom black sauce according to the present invention as a food additive. (a) and (c) were used in risotto, and (b) and (d) were used in bakery.)

[0038] The present invention is described in more detail below. However, the present invention may be implemented in various modified forms and is not limited to the implementation examples described herein.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In general, the nomenclature used herein and the experimental methods described below are well known and commonly used in the art.

[0040] The present invention provides a method for producing a mushroom melanin solution and a mushroom melanin solution produced thereby.

[0041] The method for producing the above mushroom melanin solution may include (a) a step of crushing mushrooms to obtain mushroom powder, or crushing mushrooms, mixing them with water, and then squeezing the juice with a cotton cloth or a juicer to obtain mushroom juice, and (b) a step of reacting the mushroom powder or mushroom juice with a substrate to obtain a mushroom melanin solution.

[0042] The above mushroom powder can be obtained using a mixer or the like. There is no limitation on the size of the mushroom powder, but for example, the size of the powder can be 500 μm or less.

[0043] The above mushroom melanin solution means a solution containing melanin derived from mushrooms, which is a product of reaction between mushroom powder or mushroom juice and a substrate.

[0044] The above substrate may be selected from the group consisting of amino acids such as L-tyrosine, L-cysteine, L-histidine, and L-lysine; polyphenols such as phloretin, resveratrol, P-coumaric acid, caffeic acid, isoflavonoids, flavonoids, catechol, catechin, tyrosol, tannic acid, and gallic acid; and extracts of plants containing various polyphenols (e.g., blueberry extract, tea leaf extract, soybean extract, etc.) and mixed extracts.

[0045] Preferably, the substrate may be L-tyrosine, phloretin, resveratrol, catechin, tannic acid, gallic acid.

[0046] The above mushrooms are not limited to mushrooms containing tyrosinase. Preferably, the mushrooms may be oyster mushrooms, shiitake mushrooms, oyster mushrooms, or oyster mushrooms. More preferably, they may be oyster mushrooms.

[0047] The method for producing the above mushroom melanin solution may further include, after step (b), a step (c) of low-temperature sterilization at 60 to 65°C for 25 to 35 minutes, high-temperature sterilization at 120°C or higher for 4 minutes or more, or ultraviolet (UV) sterilization for 30 minutes or more. It is more preferable that the low-temperature sterilization step be performed at 62 to 64°C for about 30 minutes.

[0048] In the above step (b), the concentration of the substrate may be tyrosine of 4 to 8 mM. More preferably, the concentration of tyrosine may be 4 mM.

[0049] In step (b), the L-tyrosine may be contained in a food. Specifically, the food may be avocado, almond, banana, cheese and other dairy products, fish, red meat, legumes, poultry, sesame seeds, pumpkin seeds, etc.

[0050] The method for producing the above mushroom melanin solution may further include, after step (c), a step (d) of concentrating by centrifugation at 6,000 to 12,000 xg (rcf) for 20 to 35 minutes.

[0051] Specifically, step (d) may be a centrifugation step at 7000 xg (rcf) for 20 minutes.

[0052] The present invention also relates to a mushroom melanin solution manufactured according to the above manufacturing method.

[0053] The above mushroom melanin solution may contain melanin or eumelanin.

[0054] Additionally, the mushroom melanin solution may further comprise a thickening agent selected from the group consisting of gellan gum, xanthan gum, carboxymethylcellulose, konjac gum, tamarind gum, guar gum, alginic acid, carrageenan, and sodium caseinate. More preferably, the mushroom melanin solution may further comprise xanthan gum as a thickening agent.

[0055] The above mushroom melanin solution may additionally contain an emulsifier selected from the group consisting of lecithin, saponin, cocobetaine, polysorbate, glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, polyoxyethylene fatty acid ester, and propylene glycol fatty acid ester.

[0056] The above mushroom melanin solution may be prepared in the form of a concentrate, paste, freeze-dried, powder or granule formulation.

[0057] In the present invention, a solution in which additives such as a thickener and an emulsifier are added to a mushroom melanin solution is defined as a mushroom black sauce, and the mushroom black sauce has a sauce formulation and can be used as a food additive.

[0058] In addition, the present invention relates to a food additive comprising the mushroom melanin solution.

[0059] The above food additive may be in the form of a sauce.

[0060] The above food additive may also be used in a food selected from the group consisting of bakery, rice cake, tofu, noodles, red pepper paste, soybean paste, soybean paste, pasta sauce and food dressing.

[0061] The present invention can control melanin production by selecting tyrosinase and substrate extracted from mushrooms, thereby controlling the properties of the product and controlling the melanin content, thereby providing a differentiated food product.

[0062] [Example]

[0063] Experimental Example 1: Method for preparing mushroom juice and mushroom melanin solution

[0064] To extract mushroom juice, 300g of shiitake mushrooms and 300g of water were ground in a 1:1 ratio in a blender. Adding water aids in the dissolution of tyrosinase within the mushrooms and increases the fluidity of the mushroom particles within the blender. After blending the mushrooms for approximately 2 minutes, a cotton cloth was used to separate large particles, yielding approximately 500g of mushroom juice.

[0065] Afterwards, L-tyrosine, a substrate for melanin formation, was added. Specifically, approximately 500 g of water was added to the extracted mushroom juice, and 4 to 8 mM of L-tyrosine was added to induce a reaction at 37°C for approximately 7 hours.

[0066] After the reaction was completed, it was sterilized at low temperature for about 30 minutes at 63°C.

[0067] Afterwards, the mushroom melanin solution was centrifuged at about 7,000 rpm for 20 minutes to concentrate it, and about 100 g of the mushroom melanin paste in concentrated form was obtained (Fig. 1).

[0068] Experimental Example 2: Mushroom Screening for Preparation of Mushroom Melanin Solution

[0069] The above manufacturing method was performed using shiitake, oyster, shiitake, and pine mushrooms. The production of mushroom melanin solution was confirmed by observing the color of the solution before and after reaction with L-tyrosine (Fig. 2). As a result, the solution turned black in shiitake mushrooms, indicating the production of mushroom melanin solution, but this reaction did not occur in other mushroom species.

[0070] Experimental Example 3: Determination of Melanin Production According to Substrate and Mushroom Concentration

[0071] Melanin production was measured over time according to the reaction concentration of the substrate L-tyrosine and the shiitake mushroom. The higher the concentration of mushrooms and L-tyrosine, the more melanin was produced, and it was confirmed that the production increased as the reaction time passed (see Fig. 3 and Table 1 below).

[0072] [Table 1]

[0073]

[0074] Experimental Example 4. Confirmation of the consumption time of L-tyrosine substrate.

[0075] In Experimental Example 3, the amount of melanin produced according to the reaction time was confirmed, but in Experimental Example 4, the time for the substrate L-tyrosine to be consumed was additionally confirmed. The substrate L-tyrosine is further oxidized to L-dopaquinone after undergoing a hydroxylation reaction to L-dopa by tyrosinase extracted from mushrooms. Afterwards, the final melanin is produced through the intermediate dopachrome. Therefore, the consumption time of the L-tyrosine substrate was confirmed by measuring the absorbance corresponding to dopachrome, the intermediate in the production process of the mushroom melanin.

[0076] For test groups 3 and 4 in Experimental Example 3 (L-tyrosine concentrations of 4 mM and 8 mM), the absorbance at a wavelength of 475 nm (the absorbance corresponding to dopachrome) was measured at 2, 4, 6, and 8 hours after the start of the reaction, respectively. As a result, in both test groups, the absorbance corresponding to 475 nm was less than 0.1 mAU at 8 hours after the start of the reaction, indicating that dopachrome was no longer produced. Therefore, it was confirmed that all tyrosine was consumed after 8 hours of reaction (Fig. 4).

[0077] Experimental Example 5. Preparation of Mushroom Melanin Solution Formulation

[0078] Various formulations of mushroom melanin solution produced through the reaction of mushroom extract and L-tyrosine were prepared. Specifically, (a) a dry formulation was prepared by removing moisture from a mushroom melanin solution using a freeze dryer, (b) a paste formulation was prepared by obtaining a solid after removing the supernatant using a centrifuge, and (c) a liquid formulation was prepared by dispersing a portion of the paste formulation in water (Fig. 5).

[0079] Experimental Example 6: Application of Mushroom Melanin Solution

[0080] Mushroom melanin solution can be used as a food additive. A representative control of the present invention is squid ink. Mushroom melanin solution can be used as a food additive in sauces, food coloring, dressings, and other foods, including gochujang (red pepper paste), soy sauce, soybean paste, bread, rice cakes, tofu, noodles, and pasta sauces.

[0081] As can be seen in Fig. 6, when the mushroom melanin solution of the present invention is added as a food additive to risotto and bakery (Fig. 6 (c) and (d) of the present invention), it can produce a color similar to that of squid ink, which is the control group (Fig. 6 (a) and (b) of the control group).

Claims

1. (a) A step of crushing mushrooms to obtain mushroom powder, or crushing mushrooms, mixing them with water, and then squeezing the juice with a cotton cloth or juicer to obtain mushroom juice, and (b) a step of reacting the mushroom powder or mushroom juice with a substrate to obtain a mushroom melanin solution, A method for producing a mushroom melanin solution, wherein the substrate is selected from the group consisting of amino acids selected from L-tyrosine, L-cysteine, L-histidine or L-lysine; polyphenols selected from phloretin, resveratrol, P-coumaric acid, caffeic acid, isoflavonoids, flavonoids, catechol, catechin, tyrosol, tannic acid or gallic acid; and an extract of a plant containing polyphenols selected from a blueberry extract, a tea leaf extract, a soybean extract or a mixed extract thereof.

2. A method for producing a mushroom melanin solution according to claim 1, wherein the mushroom is a pine mushroom, a Japanese oyster mushroom, a clam mushroom or a porous mushroom.

3. A method for producing a mushroom melanin solution, further comprising, after step (b) in paragraph 1, a step (c) of low-temperature sterilization at 60 to 65°C for 25 to 35 minutes, high-temperature sterilization at 120°C or higher for 4 minutes or longer, or ultraviolet (UV) sterilization for 30 minutes or longer.

4. A method for producing a mushroom melanin solution in step (b) of paragraph 1, wherein the substrate is 4 to 8 mM L-tyrosine.

5. A method for producing a mushroom melanin solution, wherein in step (b), the L-tyrosine is contained in a food.

6. A method for producing a mushroom melanin solution, wherein, in the third paragraph, after step (c), the method further comprises the step of concentrating by centrifugation at 6,000 to 12,000 xg (rcf) for 20 to 35 minutes (d).

7. A mushroom melanin solution manufactured according to any one of the manufacturing methods of clauses 1 to 6.

8. In paragraph 7, the mushroom melanin solution is a mushroom melanin solution containing melanin or eumelanin.

9. In paragraph 7, the mushroom melanin solution further comprises a thickening agent selected from the group consisting of gellan gum, xanthan gum, carboxymethyl cellulose, konjac gum, tamarind gum, guar gum, alginic acid, carrageenan, and sodium caseinate.

10. In paragraph 7, the mushroom melanin solution further comprises an emulsifier selected from the group consisting of lecithin, saponin, cocobetaine, polysorbate, glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, polyoxyethylene fatty acid ester, and propylene glycol fatty acid ester.

11. In paragraph 7, the mushroom melanin solution is prepared in the form of a concentrate, paste, freeze-dried, powder or granule formulation.

12. Food additive containing the mushroom melanin solution of Article 11.

13. In paragraph 12, the food additive is a food additive in the form of a sauce.

14. In the 13th paragraph, the food additive is a food additive for use in a food selected from the group consisting of bakery products, rice cakes, tofu, noodles, red pepper paste, soybean paste, soybean paste, pasta sauce, and food dressing.

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