KR102483343B1 - Sparassis crispa extract with enhanced functional components - Google Patents

Abstract

The present invention relates to a method for producing a Sparassis crispa extract, and a Sparassis crispa extract produced by the method, wherein the method comprises the steps of: repeating a drying step after spraying a natural extract on the Sparassis crispa; and performing extraction after adding water to the dried Sparassis crispa.

Description

Flower mushroom extract with enhanced functional components {Sparassis crispa extract with enhanced functional components}

The present invention comprises the steps of (1) preparing a quick-stage extract by adding water to the quick-stage, extracting, and then filtering; (2) adding water to the leaves, extracting them, and then filtering to prepare an extract; (3) spraying the quick-release extract prepared in step (1) on the zinnia mushroom; (4) drying the sprayed zinnia mushroom of step (3); (5) spraying the dried zinnia extract prepared in step (2) on the dried zinnia mushroom in step (4); (6) drying the sprayed zinnia mushroom of step (5); and (7) adding water to the dried cauliflower mushroom in step (6) and then extracting it.

Mushrooms, which have attracted the most attention among natural products effective in the treatment and prevention of diseases, have been mainly used only for food in the past. Currently, 992 species of Korean mushrooms are classified, and among them, there are about 100 edible mushrooms and 50 poisonous mushrooms. In addition, mushrooms used for medicinal purposes are reported as 162 species in 35 families and 82 genera, but the rest of the mushrooms have not yet been identified. With the improvement of living standards, interest in health promotion and anti-aging has increased in modern society, and active research is being conducted on natural substances that can be consumed as food and have antibacterial, antioxidant, and anticancer effects.

Cinnamon mushroom ( Sparassis crispa ) is native to Korea, Japan, Europe, North America, Australia, etc., and mainly grows wild on the stump of coniferous trees or the edge of dead trees. The diameter of the fruiting body is 10-30 cm in size, the height is 10-25 cm, it is fleshy, and the lower part is a stalk with a thick stem. The fruiting layer of the zinnia mushroom developed on the back of the petals, and the stalk is short, blunt, and hard with a length of 2 to 5 cm and a width of 2 to 4 cm. In addition, the tissue is white, and the spores are egg-shaped or oval with a length of 5 to 7 μm and a width of 3 to 5 μm, and the surface is flat and the spores are white. As it is known that the zinnia mushroom has a large amount of beta glucan (containing 43.6 grams per 100 grams of dry fruiting body), research on artificial cultivation methods is rapidly progressing.

Korean Patent No. 1513712 discloses a method for producing a fermented zinnia mushroom extract with increased water-soluble dietary fiber content, and Korean Patent No. 1068699 discloses a composition having antioxidant activity containing a zinnia mushroom extract. It is different from the zinnia mushroom extract in which the functional ingredient of the invention is enhanced.

The present invention has been derived from the above needs, and an object of the present invention is to improve the content of functional components and prepare a mushroom extract having excellent palatability using a mushroom mushroom rich in beta-glucan content.

In order to solve the above problems, the present invention comprises the steps of (1) preparing a quick-stage extract by adding water to the quick-stage, extracting and then filtering; (2) adding water to the leaves, extracting them, and then filtering to prepare an extract; (3) spraying the quick-release extract prepared in step (1) on the zinnia mushroom; (4) drying the sprayed zinnia mushroom of step (3); (5) spraying the dried zinnia extract prepared in step (2) on the dried zinnia mushroom in step (4); (6) drying the sprayed zinnia mushroom of step (5); and (7) adding water to the dried zinnia mushroom in step (6) and then extracting it to provide a method for producing a zinnia mushroom extract, characterized in that it is produced.

In addition, the present invention provides a flower mushroom extract prepared by the above method.

The present invention has an effect that can be usefully used in various processed foods by improving the taste for easy intake while further enhancing the content of beta-glucan, which is abundant in the mushroom mushroom, while removing the peculiar odor of the mushroom mushroom.

In order to achieve the object of the present invention, the present invention

(1) preparing a quick-stage extract by adding water to quick-stage, extracting, and filtering;

(2) adding water to the leaves, extracting them, and then filtering to prepare an extract;

(3) spraying the quick-release extract prepared in step (1) on the zinnia mushroom;

(4) drying the sprayed zinnia mushroom of step (3);

(5) spraying the dried zinnia extract prepared in step (2) on the dried zinnia mushroom in step (4);

(6) drying the sprayed zinnia mushroom of step (5); and

(7) It provides a method for producing a mushroom extract characterized in that it is prepared by adding water to the dried mushroom mushroom in step (6) and then extracting it.

In the method for producing the zinnia mushroom extract of the present invention, the quick-stage extract in step (1) is preferably extracted at 80-100 ° C. for 20-40 minutes after adding 0.8-1.2 L of water to 80-120 g of rapid-stage. It can be prepared by filtration, more preferably by adding 1 L of water to 100 g of rapid cutting, followed by extraction at 90 ° C. for 30 minutes and then filtration.

In addition, in the manufacturing method of the zinnia mushroom extract of the present invention, the fermented persimmon extract in step (2) is preferably added to 80 to 120 g of persimmon leaves with 1.8 to 2.2 L of water, followed by 2 to 4 hours at 60 to 80 ° C. It can be prepared by filtering after extraction for a period of time, more preferably by adding 2 L of water to 100 g of Korean cabbage leaves, extracting at 70 ° C. for 3 hours, and then filtering.

In addition, in the method for producing the zinnia mushroom extract of the present invention, the step (3) may preferably spray the zinnia mushroom with the rapid stage extract at a ratio of 8.8 to 9.2: 0.8 to 1.2 (w: v), more preferably can be sprayed with cauliflower mushroom at a ratio of 9:1 (w:v).

In addition, in the method for producing the zinnia mushroom extract of the present invention, the step (4) may preferably dry the sprayed zinnia mushroom at 20 to 30 ° C. for 1 to 3 days, more preferably the sprayed zinnia mushroom. can be dried for 2 days at 20-30 ° C.

In addition, in the method for producing the zinnia mushroom extract of the present invention, in the step (5), the extract may be preferably sprayed on the dried zinnia mushroom at a ratio of 8.8 to 9.2: 0.8 to 1.2 (w: v), more preferably Preferably, the dried zinnia mushroom can be sprayed with the yeokyun extract at a ratio of 9:1 (w:v).

In addition, in the method for producing the zinnia mushroom extract of the present invention, step (6) may preferably dry the sprayed zinnia mushroom at 20 to 30 ° C. for 1 to 3 days, more preferably the sprayed zinnia mushroom. can be dried for 2 days at 20-30 ° C.

In addition, in the manufacturing method of the zinnia mushroom extract of the present invention, the step (7) is preferably 8 to 12 times (v / w) of water compared to the zinnia mushroom added to the dried zinnia mushroom, and then 50 to 70 °C at 80 ~ 100 ° C. It can be extracted for minutes, more preferably, it can be extracted for 60 minutes at 90 ° C. after adding 10 times (v / w) of water compared to blossom mushrooms to dried blossom mushrooms.

The method for producing the zinnia mushroom extract of the present invention is more specifically

(1) adding 0.8 to 1.2 L of water to 80 to 120 g of rapid stage, followed by extraction at 80 to 100 ° C. for 20 to 40 minutes, followed by filtration to prepare a rapid stage extract;

(2) adding 1.8 to 2.2 L of water to 80 to 120 g of saffron leaves, extracting at 60 to 80 ° C. for 2 to 4 hours, and then filtering to prepare a saffron extract;

(3) spraying the mushroom extract prepared in step (1) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);

(4) drying the sprayed zinnia mushroom of step (3) at 20-30 ° C for 1-3 days;

(5) spraying the dried cauliflower mushroom in step (4) with the extract prepared in step (2) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);

(6) drying the sprayed zinnia mushroom of step (5) at 20-30 ° C for 1-3 days; and

(7) adding 8 to 12 times (v/w) of water compared to the zinnia mushroom to the dried zinnia mushroom in step (6) and then extracting at 80 to 100 ° C. for 50 to 70 minutes,

more specifically

(1) adding 1 L of water to 100 g of rapid stage, followed by extraction at 90° C. for 30 minutes, followed by filtration to prepare a rapid stage extract;

(2) adding 2 L of water to 100 g of Yeo Kyun leaves, extracting at 70 ° C. for 3 hours, and then filtering to prepare a Yeo Kwi extract;

(3) spraying the mushroom extract prepared in step (1) at a ratio of 9: 1 (w: v);

(4) drying the sprayed zinnia mushroom of step (3) at 20 to 30 ° C for 2 days;

(5) spraying the dried zinnia extract prepared in step (2) at a ratio of 9:1 (w:v) to the dried zinnia mushroom in step (4);

(6) drying the sprayed zinnia mushroom of step (5) at 20 to 30 ° C for 2 days; and

(7) adding 10 times (v/w) of water to the dried zinnia mushroom in step (6) and then extracting at 90° C. for 60 minutes.

The present invention also provides a flower mushroom extract prepared by the above method.

The present invention also provides a processed food containing the zinnia mushroom extract. There is no particular limitation on the type of the processed food. Examples of foods to which the zinnia mushroom extract can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, rice cakes, dairy products including ice cream, various soups, beverages, There are tea, drinks, alcoholic beverages and vitamin complexes, etc., and includes all processed foods in a conventional sense.

Hereinafter, production examples and examples of the present invention will be described in detail. However, the following Preparation Examples and Examples are only to illustrate the present invention, and the content of the present invention is not limited to the following Preparation Examples and Examples.

Preparation Example 1. Zinnia mushroom extract

(1) Sokdan ( Phlomis umbrosa ) After adding 1 L of purified water to 100 g, extraction was performed at 90 ° C. for 30 minutes, followed by filtration to prepare a rapid-stage extract.

(2) After adding 2 L of purified water to 100 g of Yeo Kyun leaves, extracting at 70 ° C. for 3 hours, and then filtering to prepare Yeo Kyun extract.

(3) The quick-release extract prepared in step (1) was evenly sprayed on the zinnia mushroom at a ratio of 9: 1 (w: v).

(4) The sprayed flower mushrooms in step (3) were naturally dried at 20-30 ° C for 2 days.

(5) The naturally dried flower mushroom in step (4) was evenly sprayed with the extract prepared in step (2) at a ratio of 9: 1 (w: v).

(6) The sprayed flower mushroom in step (5) was naturally dried at 20-30 ° C for 2 days.

(7) 10 times (v/w) of purified water compared to the zinnia mushroom was added to the naturally dried zinnia mushroom in step (6), extracted at 90 ° C. for 1 hour, and then filtered.

Comparative Example 1. Zinnia mushroom extract

(1) Cauliflower mushrooms were naturally dried at 20-30 ° C for 2 days.

(2) 10 times (v/w) of purified water compared to the zinnia mushroom was added to the naturally dried zinnia mushroom in step (1), extracted at 90 ° C. for 1 hour, and then filtered.

Comparative Example 2. Zinnia mushroom extract

(1) Sokdan ( Phlomis umbrosa ) After adding 1 L of purified water to 100 g, extraction was performed at 90 ° C. for 30 minutes, followed by filtration to prepare a rapid-stage extract.

(2) The quick-release extract prepared in step (1) was evenly sprayed on the zinnia mushroom at a ratio of 9: 1 (w: v).

(3) The sprayed flower mushrooms in step (2) were naturally dried at 20 to 30 ° C for 2 days.

(4) After adding 10 times (v/w) of purified water compared to the zinnia mushroom to the naturally dried zinnia mushroom in step (3), extraction was performed at 90 ° C. for 1 hour and filtered.

Comparative Example 3. Zinnia mushroom extract

(1) After adding 2 L of purified water to 100 g of Yeo Kyun leaves, extracting at 70 ° C. for 3 hours, and then filtering to prepare Yeo Kyun extract.

(2) The mushroom leaf extract prepared in step (1) was evenly sprayed on the flower mushroom at a ratio of 9: 1 (w: v).

(3) The sprayed flower mushrooms in step (2) were naturally dried at 20 to 30 ° C for 2 days.

(4) After adding 10 times (v/w) of purified water compared to the zinnia mushroom to the naturally dried zinnia mushroom in step (3), extraction was performed at 90 ° C. for 1 hour and filtered.

Experiment method

1. Measurement of β-glucan content

(a) Pretreatment for total glucan analysis: 0.1 g of each lyophilized sample was taken in a cap tube, 2 mL of 12M H ₂ SO ₄ was added, the cap was closed, and the mixture was vortexed from time to time at 4 ° C. Digested for 2 hours. After carefully adding 4 mL of distilled water to the degraded sample and vortexing for 10 seconds, 6 mL of distilled water was added and further vortexing for 10 seconds. After that, the cap was slightly opened and reacted in boiling water (100 ° C.) for 5 minutes, and then the lid was closed and reacted for an additional 2 hours. After the reaction was completed, the sample was slowly cooled at room temperature, the cap was carefully opened, taken into a 100 mL constant-use flask, and 6 mL of 10M KOH was added thereto, followed by sodium acetate buffer (200 mM, pH 5). After filtering, it was used as a total glucan test solution.

(b) Analysis of α-glucan and free glucose Pretreatment: 0.1 g of each lyophilized sample was taken in a cap tube, 2 mL of 2M KOH was added, stirred, and allowed to stand at 4° C. for 20 minutes. Then, 8 mL of sodium acetate buffer (1.2M, pH 3.8) was added, and 0.2 mL of bottle 2 (Amyloglucosidase (1,630 U/mL) plus invertase (500 U/mL) solution in 50% (v/v) glycerol) was added. and then reacted at 40° C. for 30 minutes. Then, 100 mL of distilled water was added, filtered, and used as a test solution.

(c) Total glucan analysis: Take 0.1 mL of test solution and take 0.1 mL of bottle 1 (exo-1,3-β-Glucanase (100 U/mL) plus β-Glucosidase (20 U/mL) ammonium sulphate suspension) After vortexing, the mixture was reacted at 40° C. for 60 minutes. Thereafter, 3 mL of GOPOD reagent was added and reacted at 40° C. for 20 minutes, and then absorbance was measured at 510 nm. As a reagent blank, 0.2 mL sodium acetate buffer (200 mM, pH 5.0) and 3 mL GOPOD reagent were used, and as glucose standards, 0.1 mL sodium acetate buffer (200 mM, pH 5.0) and 0.1 mL D- A glucose standard solution (1.00 mg/mL in 0.2% benzoic acid) and 3 mL GOPOD reagent were used.

(d) Analysis of α-glucan and free glucose: 0.1 mL of the test solution was taken, 0.1 mL of sodium acetate buffer (200 mM, pH 5.0) was vortexed, 3 mL of GOPOD reagent was added, and 40 ° C. After reacting for 20 minutes, absorbance was measured at 510 nm. As a reagent blank, 0.2 mL sodium acetate buffer (200 mM, pH 5.0) and 3 mL GOPOD reagent were used, and as glucose standards, 0.1 mL sodium acetate buffer (200 mM, pH 5.0) and 0.1 mL D- A glucose standard solution (1.00 mg/mL in 0.2% benzoic acid) and 3 mL GOPOD reagent were used.

2. Anti-inflammatory activity

(a) Raw 264.7 macrophage cell line pre-treatment

Experiments were conducted using rat macrophage Raw 264.7 cells. Raw 264.7 cells were cultured using DMEM medium supplemented with 10% FBS and 1% antibiotics (penicillin/streptomycin), and were subcultured before reaching confluent and used for experiments.

(b) Evaluation of viability of Raw 264.7 macrophage cell line

The cell viability of Raw 264.7 cell samples was examined for toxicity using cell counting kit-8, and the concentration of the sample to be used for anti-inflammatory evaluation was determined through this. Raw 264.7 cells in culture were seeded in a 96-well plate at 5×10 ⁴ cells/mL, cultured for 24 hours in an incubator at 37° C. and 5% CO ₂ , and samples were then diluted to a certain concentration and treated. After incubation for 48 hours, the medium was removed, and 100 μl of PBS and 10 μl of cck-8 reagent were added thereto, followed by reaction in a dark room at 37° C. and 5% CO ₂ incubator for 2 hours. The absorbance of each well developed was measured at 450 nm using a microplate reader, and the cell viability was confirmed using the following formula.

Cell viability (%) = sample absorbance/control absorbance × 100

(c) Comparison of production of NO (Nitric oxide)

The amount of NO produced from Raw 264.7 cells was measured using Grease reagent in the form of NO ₂ . Raw 264.7 cells in culture were seeded in a 96-well plate at 5×10 ⁵ cells/mL, cultured in an incubator at 37° C. and 5% CO ₂ for 24 hours, and then treated with a dilution factor that maintained a cell viability of 80% or more and LPS. (1 μg/mL) was divided into experimental groups and non-added groups and cultured for up to 24 hours. After incubation, 100 μl of the culture supernatant was mixed with 100 μl of grease reagent and allowed to react at room temperature for 10 minutes. After the reaction, absorbance was measured at 540 nm using a microplate reader, and NO production was measured with uM nitrite using sodium nitrite (NaNO ₂ ) as a standard material.

Example 1. β-glucan content

Glucan is a polysaccharides and is contained in a large amount in mushrooms and grains such as barley, oats, and rye. Although it has excellent anticancer effects, it does not directly kill cancer cells, but increases immunity, suppresses the activity of cancer cells, and increases the number and activity of cells related to immune function, such as T cells. In addition, β-glucan is a type of polysaccharide that is abundantly contained in mushrooms, cell walls of yeast, grains, and fungi. The results of measuring the beta-glucan content of the zinnia mushroom extract are shown in Table 1 below.

β-glucan content of zinnia mushroom extract extract type β-glucan content (%) Preparation Example 1 6.7±0.1 Comparative Example 1 4.5±0.0 Comparative Example 2 5.2±0.2 Comparative Example 3 5.6±0.1

As a result, it was confirmed that the zinnia mushroom extract extracted by the method of Preparation Example 1 had the highest β-glucan content.

Example 2. Anti-inflammatory activity

1. Cell viability

As a result of evaluating the cell viability of RAW 264.7 macrophages, all four samples at a concentration of 1 mg/mL showed a viability of 95% or higher (Table 2). Therefore, it was prepared at a concentration of 1 mg/mL and the following experiment was conducted.

Cytotoxicity of zinnia mushroom extract extract type Cell viability (%) Preparation Example 1 98.2±0.8 Comparative Example 1 96.3±0.6 Comparative Example 2 95.4±1.1 Comparative Example 3 97.2±1.6

2. Macrophage NO (nitric oxide) production

NO (nitric oxide), which plays a role in immune activity, is an inorganic low-molecular-weight radical, and is known to play a role in nerve transmission, blood coagulation and blood pressure control, and immune function against cancer cells. synthesized from In particular, macrophages (macrophase) produce NO (nitric oxide) by stimuli such as inflammation in vivo to play an important role in immune activity such as anti-inflammatory and anti-tumor.

Inflammation was induced using 1 μg/mL of LPS (Lipopoly saccharide), which is mainly used as an inflammation-inducing substance, using immune cells, and NO ₂ ^- (nitrite), a NO oxide generated from the cells of Raw 264.7, was mixed with a grease reagent. The reaction color was developed to indicate the degree of macrophage generation. In the LPS treatment group, the amount of NO production increased, but when the sample was diluted and treated at a concentration of 1 mg/mL, the amount of NO production decreased, and among the extracts, the extract of Preparation Example 1 decreased the most.

Amount of nitric oxide (NO) produced by the zinnia mushroom extract extract type NO production amount (uM) DMEM 0.2±0.0 DMEM+LPS 21.2±1.2 Preparation Example 1 7.1±0.9 Comparative Example 1 11.0±0.5 Comparative Example 2 9.2±1.7 Comparative Example 3 10.5±1.1

Claims (5)

(1) preparing a quick-stage extract by adding water to quick-stage, extracting, and filtering;
(2) adding water to the leaves, extracting them, and then filtering to prepare an extract;
(3) spraying the quick-release extract prepared in step (1) on the zinnia mushroom;
(4) drying the sprayed cauliflower mushroom of step (3);
(5) spraying the dried zinnia extract prepared in step (2) on the dried zinnia mushroom in step (4);
(6) drying the sprayed zinnia mushroom of step (5); and
(7) A method for producing a zinnia mushroom extract, characterized in that it is prepared by adding water to the dried zinnia mushroom in step (6) and then extracting it. According to claim 1,
(1) adding 0.8 to 1.2 L of water to 80 to 120 g of rapid stage, followed by extraction at 80 to 100 ° C. for 20 to 40 minutes, followed by filtration to prepare a rapid stage extract;
(2) adding 1.8 to 2.2 L of water to 80 to 120 g of saffron leaves, extracting at 60 to 80 ° C. for 2 to 4 hours, and then filtering to prepare a saffron extract;
(3) spraying the quick-release extract prepared in step (1) on the zinnia mushroom;
(4) drying the sprayed zinnia mushroom of step (3);
(5) spraying the dried zinnia extract prepared in step (2) on the dried zinnia mushroom in step (4);
(6) drying the sprayed zinnia mushroom of step (5); and
(7) A method for producing a zinnia mushroom extract, characterized in that it is prepared by adding water to the dried zinnia mushroom in step (6) and then extracting it. According to claim 2,
(1) adding 0.8 to 1.2 L of water to 80 to 120 g of rapid stage, followed by extraction at 80 to 100 ° C. for 20 to 40 minutes, followed by filtration to prepare a rapid stage extract;
(2) adding 1.8 to 2.2 L of water to 80 to 120 g of saffron leaves, extracting at 60 to 80 ° C. for 2 to 4 hours, and then filtering to prepare a saffron extract;
(3) spraying the mushroom extract prepared in step (1) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);
(4) drying the sprayed zinnia mushroom of step (3);
(5) spraying the dried cauliflower mushroom in step (4) with the extract prepared in step (2) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);
(6) drying the sprayed zinnia mushroom of step (5); and
(7) A method for producing a zinnia mushroom extract, characterized in that it is prepared by adding 8 to 12 times (v / w) of water compared to the zinnia mushroom to the dried zinnia mushroom in step (6) and then extracting. According to claim 3,
(1) adding 0.8 to 1.2 L of water to 80 to 120 g of rapid stage, followed by extraction at 80 to 100 ° C. for 20 to 40 minutes, followed by filtration to prepare a rapid stage extract;
(2) adding 1.8 to 2.2 L of water to 80 to 120 g of saffron leaves, extracting at 60 to 80 ° C. for 2 to 4 hours, and then filtering to prepare a saffron extract;
(3) spraying the mushroom extract prepared in step (1) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);
(4) drying the sprayed zinnia mushroom of step (3) at 20-30 ° C for 1-3 days;
(5) spraying the dried cauliflower mushroom in step (4) with the extract prepared in step (2) at a ratio of 8.8 to 9.2:0.8 to 1.2 (w:v);
(6) drying the sprayed zinnia mushroom of step (5) at 20-30 ° C for 1-3 days; and
(7) It is characterized in that it is prepared by adding 8 to 12 times (v / w) of water compared to the zinnia mushroom to the dried zinnia mushroom in step (6) and then extracting at 80 to 100 ° C. for 50 to 70 minutes Method for producing a flower mushroom extract to do. delete