KR20180045658A - Solid fermented soybean with mushroom and method for preparation thereof - Google Patents

Abstract

The present invention relates to fermented soybean which is solid-fermented by mushroom, composed of: steamed soybean; and at least one of mushroom mycelium selected among Polyozellus multiplex (Underw.) Murrill, pine mushroom, and Grifola frondosa (Dicks.) Gray, and a cultured product thereof. The present invention further relates to a production method thereof. According to the present invention, amountd of conjugated linoleic acid and non-glycoside-isoflavin are increased while content of ammonia is decreased, and flavor is improved, thereby enabling production of functional foods having antioxidative activities and effects of ameliorating diabetes and obesity.

Description

FIELD OF THE INVENTION The present invention relates to a fermented soybean fermented by a mushroom and a method for producing the fermented soybean fermented soybean,

The present invention relates to soybean fermented by solid fermentation by mushroom and a method for producing the same. More specifically, the present invention relates to a soybean fermented product obtained by solid-fermenting mushroom with improved conjugated linoleic acid and non-glycoside-isoflavin, reduced ammonia content and improved flavor, and a process for producing the same.

Soybean [ Glycine max (L.) Merrill ] has been an important part of our diet for a long time, after rice and rice, originating in the northern part of the Korean Peninsula and Manchuria. Soybean has played a very important role as a source of protein and fat in the Korean rice dish, which is made from grains such as rice and barley. Especially, soybean is the main ingredient of soybean paste, soybean paste, kochujang, and chungkukjang, which is frequently used by Korean people. It is an edible crop that is used frequently as a raw material for processed products such as soybean milk, tofu and cooking oil.

In addition, mushroom is defined as a generic name of a flock that forms visually identifiable fruiting bodies among fungi. They are composed of various nutrients such as carbohydrates, lipids, proteins, minerals and vitamins. They are characterized by their unique texture and fragrance. They are used for edible and medicinal purposes. Many mycelium and fruiting bodies of bacterium have been reported to have various anti-cancer effects and immunity enhancement. Mycelium of basidiomycetes does not have any side effects unlike the commonly used anticancer therapies, and it has the greatest advantage of enhancing immune function and exhibiting anticancer effect.

On the other hand, conjugated linoleic acid (CLA) is an isomer of linoleic acid and has a conjugated double bond structure. Linoleic acid has a cis-shaped double bond at 9 and 12 carbons, whereas the double bond position of conjugated linoleic acid is 9 and 11, 10 and 12 and 11 and 13, and the double bond type has cis-cis , cis-trans, trans-cis, trans-trans and the like, and a total of twelve conjugated linoleic acid isomers are known.

The conjugated linoleic acid (CLA) is chemically synthesized or produced by microorganisms. When chemically synthesized, it is possible that all of the isomers of conjugated linoleic acid (CLA) are produced, but cis-9 and trans-11 conjugated linoleic acid (CLA) are mainly produced by microorganisms. Conjugated linoleic acid (CLA) is present in trace amounts because meat derived from rumen contains very low concentrations of conjugated linoleic acid (CLA), and lactic acid fermented foods, especially yogurt, use skimmed milk.

In addition, there is no conjugated linoleic acid (CLA) in soy-related foods such as soy milk. The production of conjugated linoleic acid (CLA) through liquid fermentation of defatted soybean using some edible mushroom mycelium has been reported and the production of conjugated linoleic acid by soybean fermentation using fungi (fungi) has been reported, and the solid fermentation method using edible mushroom mycelium or fungus The production of conjugated linoleic acid (CLA) by a microorganism is insignificant.

In addition to nutritional ingredients, soy contains isoflavones, known as vegetable estrogens, which are widely known to be effective in preventing various adult diseases. It is known that isoflavones vary slightly depending on the soybean varieties, but generally glycosides 25%, malonyl-glycosides 70-80%, acetyl-glycosides 5% and aglycone 2% . Although several groups of isoflavones exhibit physiological activity, in particular, isoflavones of non-glycosides have been reported to have high bioavailability. These soybean varieties may vary in content of isoflavones depending on the soybean variety, cultivation environment, germination, processing, and action of microorganisms. In general, the conversion of glycosides to non-glycosyltransferases is caused by the increase, heat treatment or germination and fermentation treatment. In addition, fermented foods are degraded by β-glycosidase activity produced by fermenting microorganisms and converted to non-glycoside forms. However, the typical fermentation method using solid fermentation method requires a more efficient solid fermentation method with less than 40% of non-glycoside production rate. However, it is limited in bacteria such as Bacillus subtilis and has a low preference due to its unique smell.

As a background of the present invention, Korean Patent Laid-Open No. 10-2009-0081598 describes a method for producing fermented soybean powder solution.

As a result of continuous research in the related art, the inventors of the present invention have found that soybean fermented product produced by soybean solid fermentation using mushroom such as shrike mushroom has an increased content of conjugated linoleic acid (CLA) and non-glycosylated isoflavone and antioxidant activity, And the fermentation odor such as ammonia is reduced as compared with that of Chungkukjang. Thus, the present invention has been completed.

Accordingly, an object of the present invention is to provide a fermented soybean which is solid fermented by mushroom, wherein the content of conjugated linoleic acid and non-glycoside isoflavone is increased.

Another object of the present invention is to provide a high-quality soybean fermentation product having reduced ammonia content, improved flavor, and improved functionality such as antioxidant activity, diabetes and obesity improving effect.

It is still another object of the present invention to provide a food comprising the soybean fermented product.

It is another object of the present invention to provide a solid fermentation method of soybeans capable of increasing the non-glycoside production rate.

It is still another object of the present invention to provide a method for producing a soybean fermented product capable of economically and efficiently producing the high-quality soybean fermented product.

The object of the present invention is not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood from the description of the detailed description.

According to one aspect of the present invention, there is provided a method for producing a mushroom mycelium comprising at least one or more of at least one mushroom mycelium selected from magpie mushroom, pine mushroom, and mushroom and a culture thereof; And fermented soybean fermented solid by mushroom, including soybeans added.

According to one embodiment of the present invention, the soybean fermentation product of the present invention can increase at least one of the contents of the conjugated linoleic acid and the non-glycoside isoflavone.

According to one embodiment of the present invention, the fermented soybean of the present invention can reduce the ammonia content and improve the flavor.

According to one embodiment of the present invention, the beans may be soaked beans or germinated beans.

According to another aspect of the present invention, there is provided a food comprising the soybean fermented product.

According to one embodiment of the present invention, the food is selected from a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, a tablet, Or the like.

According to one embodiment of the present invention, the food may be a functional food having antioxidative activity, diabetes or obesity prevention or improvement effect.

According to still another aspect of the present invention, there is provided a method for producing soybean mushroom, comprising the steps of: inoculating a soybean which has been cultivated with at least one of mushroom mycelia selected from magpie mushroom, mushroom mushroom, and mushroom and a culture thereof; And a fermentation step of fermenting the soybeans.

According to still another aspect of the present invention, there is provided a method for producing soybean mushroom, comprising the steps of: inoculating a soybean which has been cultivated with at least one of mushroom mycelia selected from magpie mushroom, mushroom mushroom, and mushroom and a culture thereof; And a fermentation step of fermenting the soybean.

According to an embodiment of the present invention, the seeding step may further include soaking or germinating the soybeans before the soybean is grown.

According to one embodiment of the present invention, in the step of inoculation, the mushroom mycelium or the culture thereof may be inoculated at 5 to 10 parts by weight with respect to the weight of the soybean.

According to an embodiment of the present invention, the mushroom may be cultured at room temperature for 4 to 6 days before the inoculation step.

According to an embodiment of the present invention, the fermentation step may be performed at 20 to 30 ° C for 5 to 10 days.

According to an embodiment of the present invention, the method may further include a step of drying and pulverizing after the fermentation step.

According to an embodiment of the present invention, the pulverizing step may be performed by drying the fermented soybeans at 45 to 55 ° C for 2 to 3 days.

According to an embodiment of the present invention, after the fermentation step, the fermented soybean may be used as a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, , And < RTI ID = 0.0 > formulating < / RTI >

The soybean fermented product according to an embodiment of the present invention can increase the content of the conjugated linoleic acid and the non-glycoside isoflavone.

The soybean fermented product according to an embodiment of the present invention may be processed into various types of foods such as an alginate type powder, granule, pills, tablet, and beverage, because the fermented soybean and ammonia contents are reduced and the flavor is improved.

The soybean fermented product according to one embodiment of the present invention can provide a functional food having improved functionality such as antioxidant activity, diabetes and obesity improving effect.

The solid fermentation method of soybean according to one embodiment of the present invention can increase the non-glycoside production rate to 40% or more.

The method of manufacturing a soybean fermented product according to an embodiment of the present invention can economically and efficiently produce a high quality soybean fermented product in which the content of conjugated linoleic acid and nonisaccharide isoflavone is increased.

FIG. 1 is a photograph of soybean fermented product of an edible mushroom mycelium produced according to an embodiment of the present invention. Fig. 1 (A) is a photograph of soybeans before fermentation, B is photograph of soybean fermented product of mycelium of mushroom mycelium, C is photograph of soybean fermentation product of mycelia of leaf mushroom, and D is photograph of soybean fermentation product of black mushroom mycelium.
FIG. 2 is a GC chromatogram of conjugated linolenic acid of soybean fermentations prepared according to one embodiment of the present invention. 2 shows GC chromatogram of soybeans before fermentation, B shows GC chromatogram of soybean fermented mycelia, C shows GC chromatogram of fermented soybean mycelia, and D shows black pepper mushroom GC chromatogram of fermented soybean mycelium.
FIG. 3 is a photograph showing soaked beans and sprouted beans according to an embodiment of the present invention. Fig. 3 (A) shows soaked soybeans, and B shows germinated soybeans.
4 is a photograph of a fermented soybean powder according to the prior art and a bean fermented product powder of mycelia of edible mushrooms according to an embodiment of the present invention. FIG. 4A shows chungkukjang powder, B shows the powder of soaked soybean fermented by the magpie mushroom, and C shows the fermented powder of germinated soybean by the magpie mushroom mycelium.
5 is isoflavone HPLC chromatogram of soybean fermentations prepared according to one embodiment of the present invention. FIG. 5A shows the isoflavone HPLC chromatogram of the chrysanthemum powder, B shows the isoflavone HPLC chromatogram of the soaked fermented soybean powder with the mycelium of magpie, and C shows the fermented soybean fermented product Isoflavone HPLC chromatogram of the powder.
FIG. 6 is a chromatogram of free amino acid and ammonia amino acid automatic analyzer of soybean fermentations prepared according to an embodiment of the present invention. FIG. 6A shows the chromatogram of the free amino acid and ammonia amino acid analyzer of the chungkukjang powder, B shows the chromatogram of free amino acid and ammonia amino acid automatic analyzer of the soaking fermented soybean powder with the mycelium of magpie mushroom, The chromatogram of free amino acid and ammonia amino acid analyzer of germinated fermented soybean powder by mushroom mycelium is shown.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise.

In this application, the terms "comprises", "having", and the like are intended to specify the presence of stated features, integers, steps, operations, components, parts, or combinations thereof, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.

In the present application, when a component is referred to as " comprising ", it means that it can include other components as well, without excluding other components unless specifically stated otherwise.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention is capable of various modifications and various embodiments, and particular embodiments are illustrated in the drawings and will be described in detail in the detailed description. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. In the following description, well-known functions or constructions are not described in detail since they would obscure the invention in unnecessary detail.

According to one aspect of the present invention, there is provided a method for producing a mushroom mycelium comprising at least one or more of at least one mushroom mycelium selected from magpie mushroom, pine mushroom, and mushroom and a culture thereof; And fermented soybean fermented solid by mushroom, including soybeans added.

The soybean fermented product of the present invention can increase at least one of the contents of the conjugated linoleic acid and the non-glycoside isoflavone. In particular, the content of the non-glycoside isoflavone can be increased to 40% or more.

The Polyozellus multiplex is 5 to 15 cm in width and 5 to 30 cm in height. It is known as an edible mushroom that grows in coniferous forests such as pine trees and broad-leaved forests in Gangwon Province. In some parts of Gangwon-do, it is known as "mushroom", and in Yangyang area it is also known as "rubber mushroom" or "bear mushroom". It is known that magpie mushroom has the effect of preventing stomach cancer, anticancer efficacy and preventing dementia. According to the present invention, the mycelia or culture thereof of the black mushroom can increase the content of conjugated linoleic acid and / or non-glycoside isoflavone when used for solid fermentation of soybean. The magpie mushroom is superior to other mushrooms in producing conjugated linoleic acid.

The above mushroom ( Tricholoma matsutake ) is a mushroom that grows in the roots of red pines and forms mycorrhiza in the roots of red pines. Every year in the autumn (about 19 ℃), mycelium grows and grows suddenly. It grows suddenly and starts appearing on the ground about two weeks later, forming fruiting body. Pine mushroom is thick, fragrant, and delicious wild mushroom. It is known to have medicinal value such as edible and anti-cancer. According to the present invention, the mycelium of pine mushroom or the culture thereof can increase the content of conjugated linoleic acid and / or non-glycoside isoflavone when used for solid fermentation of soybean.

The leaf mushroom ( Grifola frondosa ) is a group of leaf-like glands that are folded in layers to form a bundle of fruit bodies. The colors are black, dark brown, grayish brown, and white. Leaf mushrooms are known to have good anticancer effects and tumor suppression rates because they have good taste and good pharmacological action. According to the present invention, the mycelium of the mushroom or the culture thereof can increase the content of conjugated linoleic acid and / or non-glycoside isoflavone when used for solid fermentation of soybean.

In the present invention, beans are meant to include all the fruits obtained from plants classified as beans, regardless of their kind. Examples include, but are not limited to, peas, beans, seomyeotae, chungtae, eastern, bamboo, white beans, herb beans, rice bran, soybean, seaweed and soybean.

In the present invention, the soybean fermented product means that fermented soybean is produced by using the fermentation action of a microorganism using soybean as a raw material.

According to one embodiment of the present invention, the fermented soybean of the present invention can reduce the ammonia content and improve the flavor. The soybean fermented product produced by the present invention has reduced ammonia content and improved flavor as compared with chungkukjang fermented by solid fermentation method using Bacillus subtilis.

According to one embodiment of the present invention, the beans may be soaked beans or germinated beans, although not limited thereto. When the soya bean or sprouts are used, the content of conjugated linoleic acid and / or non-glycoside isoflavone can be increased.

According to another aspect of the present invention, there is provided a food comprising the soybean fermented product.

As described above, the soybean fermented product produced by the present invention can be used as a food by increasing at least one of the contents of the conjugated linoleic acid and the non-glycoside isoflavone, reducing the ammonia content and improving the flavor, It can also be used as a food additive such as confectionery and beverage.

According to an embodiment of the present invention, the food is limited to a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, a tablet, And a liquid phase.

According to one embodiment of the present invention, the food may be a functional food having antioxidative activity, diabetes or obesity prevention or improvement effect. The soybean fermented product according to the present invention can be utilized as a functional food for preventing or improving antioxidant activity, diabetes or obesity when at least one of the contents of at least one of conjugated linoleic acid and non-glycoside isoflavone is increased as a food or food additive.

In the present invention, the antioxidant activity means an activity of eliminating, suppressing or suppressing the harmfulness of singlet oxygen, superoxide anion, superoxiradical or hydrogen peroxide. In the following examples of the present invention, the DPPH radical scavenging activity was verified.

The soybean fermented product according to the present invention is not limited to the soybean fermented product but may be powdered and added to the raw material mixture of cotton, bread or pastry, kneaded, and shaped into a predetermined shape to prepare cotton, bread or confection. Mixtures of ingredients commonly used in the art can be used to make such surfaces, breads or confections. In the production of the surface, bread, or confection, the amount of soybean fermentation product added can be up to 99.99%.

According to still another aspect of the present invention, there is provided a method for producing soybean mushroom, comprising the steps of: inoculating a soybean which has been cultivated with at least one of mushroom mycelia selected from magpie mushroom, mushroom mushroom, and mushroom and a culture thereof; And a fermentation step of fermenting the soybeans.

According to the solid fermentation method of soybean of the present invention, at least one of the contents of the conjugated linoleic acid and the non-glycoside isoflavone of the fermented soybean can be increased. In particular, the content of the non-glycoside isoflavone can be increased to 40% or more. Further, according to the solid fermentation method of soybean of the present invention, the soybean fermented product has a reduced ammonia content and an improved flavor as compared with chungkukjang fermented by solid fermentation method with Bacillus subtilis.

According to still another aspect of the present invention, there is provided a method for producing soybean mushroom, comprising the steps of: inoculating a soybean which has been cultivated with at least one of mushroom mycelia selected from magpie mushroom, mushroom mushroom, and mushroom and a culture thereof; And a fermentation step of fermenting the soybean.

According to the method for producing a soybean fermented product of the present invention, at least one of the contents of the conjugated linoleic acid and the unglycosylated isoflavone of the soybean fermented product can be increased. In particular, the content of the non-glycoside isoflavone can be increased to 40% or more. Further, according to the method for producing soybean fermented product of the present invention, the soybean fermented product has reduced ammonia content and improved flavor as compared with chungkukjang, which is fermented by solid fermentation method using Bacillus subtilis.

In the present invention, there is no particular limitation as long as the soy sauce is boiled so as to induce sterilization and fermentation and sterilized unless the nutritional ingredients are destroyed. For example, at 120 ° C for 30 to 60 minutes. If the sterilization time is less than 30 minutes, the soybean may not be boiled or sterilized and the fermentation may not proceed. If the sterilization time is more than 60 minutes, excessive disinfection and nutrients may be lost.

According to an embodiment of the present invention, the seeding step may further include soaking or germinating the soybeans before the soybean is grown. There is no particular limitation on the condition of soaking if the soaking provides sufficient moisture to the soybean. For example, three times as much water as beans can be added to the flour and the flour can be soaked at room temperature for 8-12 hours. The germination initiates the generation or growth of the bud of the soybean, and the germination condition is not particularly limited. For example, germinated soybeans can be produced by sprouting two times as much of the soaked soybeans.

According to one embodiment of the present invention, in the step of inoculation, the mushroom mycelium or the culture thereof may be inoculated at 5 to 10 parts by weight with respect to the weight of the soybean. If the mushroom mycelium or the cultured product thereof is less than 5 parts by weight based on the soybean weight, the fermentation rate is delayed, and if it exceeds 10 parts by weight, there is no economical efficiency irrespective of the fermentation rate.

According to an embodiment of the present invention, the step of culturing the mushroom at room temperature for 5 to 15 days prior to the seeding step may further include providing seeds. The seed culture of the mushroom can be cultured using known techniques known in the art. For example, rice bran can be washed with water to prepare a liquid broth, and the broth can be cultured at room temperature for 5 to 15 days using the liquid broth.

According to an embodiment of the present invention, the fermentation step is preferably performed at 20 to 30 ° C for 6 to 12 days, but is not limited thereto. When the fermentation temperature is lower than 20 캜, the fermentation rate is retarded. When the temperature is higher than 30 캜, the growth of the mycelium is inhibited and the fermentation rate is remarkably increased. If the fermentation period is less than 6 days, the mycelial growth is insufficient, and if it is more than 8 days, the active ingredient etc. may decrease.

According to an embodiment of the present invention, the method may further include a step of drying and pulverizing after the fermentation step. The drying may be performed at 45 to 55 ° C. for 2 to 3 days. If the drying temperature is less than 45 ° C, the drying period becomes longer and contamination of Bacillus subtilis may occur. When the drying temperature exceeds 55 ° C, the nutrients may rapidly break down. If necessary, freeze-drying can be performed to minimize the destruction of nutritional ingredients. The dried beans may be pulverized by a pulverizer to be provided as a powder.

According to an embodiment of the present invention, after the fermentation step, the fermented soybean may be used as a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, , And < RTI ID = 0.0 > formulating < / RTI > The above-described formulation method can be carried out using a known technique in the art.

The soybean fermented product of the mushroom mycelium according to the present invention, particularly the soybean fermented product of the magpie mushroom mycelium, is excellent in the production ability of the conjugated linoleic acid (see Test Examples 1 to 2 and Example 1) , The content of conjugated linoleic acid and non-glycoside-isoflavone is higher than that of cheonggukjang prepared by the solid fermentation method. Also, since the content of ammonia, which is a unique protein degradation product, is low, the odor of the fermentation odor is remarkably low and the flavor is improved. Test Example 1 and Examples 1 to 2). In addition, the antioxidative activity and the improvement effect of diabetes and obesity are remarkably improved, and the functionalities are also excellent (see Test Example 1 and Examples 1 to 2).

According to another object of the present invention, there is provided a high-quality soybean fermented milk powder, a granule, a granule, a flavor enhancer, an antioxidant activity improving effect, an improvement in diabetes and obesity, Pills, tablets and beverages.

The present invention will be described in more detail by the following examples. These examples are for illustrating the present invention, and the scope of the present invention should not be limited by them.

Example  1 to 3: of edible mushroom mycelium Conjugated linoleic acid Production capacity  Confirm

The amount of conjugated linoleic acid (CLA) was determined from three mycelia of mushroom (pine mushroom, mushroom and mushroom).

Test Example 1

i) Production of fermented seeds

After washing with water for 12 hours, the water was removed, and 10 g was weighed into a 250 mL Erlenmeyer flask. Sterilized water (90 mL) was added and sterilized at 121 ° C for 15 minutes to prepare a 10% rice liquid medium.

Each of the plates cultivated with mycelium of matsutake mushroom, mushroom mycorrhiza mycelia and mushroom mycelia was inoculated aseptically into 10% rice liquid medium and cultured at 25 ° C for 10 days to prepare seeds for fermentation.

Mushroom mycorrhizae were collected from the nearby mountains of Anyeong - myeon, Hamyang - gun, Gyeongsangnam - do, The cultivation of mycorrhizal mushroom mycelium was carried out in a potato dextrose (PDA, Difico, MI, USA) solid medium for 10 days, once every 2-3 months.

ii) Increase soybean  Produce

3 kg of yellow soybeans were washed with flowing water, and water was removed. After water was dipped in 20 ° C for 12 hours, the water was removed. 200 g of soaked soybean was divided into polystyrene fermentation vessels, and the soybean was sterilized by heating at 121 ° C for 30 minutes.

iii) Fermentation

Each sterilized soybean was inoculated with 5% (w / v) each of the prepared seed culture and fermented at 25 ° C for 12 days (Fig. 1).

iv) Soybean fermentation product  Powder manufacturing

The fermented soybean fermented product was dried at 50 DEG C for 3 days, pulverized by a pulverizer, and pulverized.

≪ Analysis of conjugated linolenic acid &

Analysis of the conjugated linoleic acid was carried out by extracting 2 g of the powder and extracting fatty acids by adding chloroform: methanol (1: 1) to the fatty acid analysis. That is, the fatty acid analysis was carried out according to the fatty acid analysis method of the food method.

Before the fatty acid analysis, the fat content of the raw materials of each soybean variety was extracted. That is, 30 mL of a chloroform: methanol (2: 1) Folch solution was added to 10 g of soybean powder for each breed, and the fat was extracted by stirring at room temperature for 2 hours.

After the extraction of fat, the filtrate was filtered and 4 mL of saturated NaCl was added to the filtrate. Only the fat layer with layer separation was recovered again, dehydrated with anhydrous sulfuric acid, and concentrated under reduced pressure at 60 ° C to recover pure fat.

To each 1 mL of soybean fat obtained, 3 mL of 0.5 N methanolic NaOH was added and heated at 100 ° C for 10 minutes to hydrolyze the fatty acid and glycerol. Then, 2 mL of boron trifluoride (BF ₃ ) was added and stirred, and the mixture was heated again for 30 minutes to proceed with fatty acid methyl esterification.

After the reaction was completed, 1 mL of isooctane was added, and the mixture was vigorously shaken, centrifuged, and the separated iso-octane layer was collected, dehydrated with anhydrous sodium sulfate, and filtered through a 0.45 μm-membrane filter (Dismic-25CS).

As a result, as shown in Fig. 2, no conjugated linolenic acid was detected in the soybeans before fermentation (Comparative Example 1), and the total conjugated linolenic acid content of the soybean mushroom mycelium and the mushroom mycelia after fermentation of the mycelia of the mushrooms of Examples 2 and 3 was about 20 To 25 μg / g, and in the case of bean (Example 3) after the fermentation of the mushroom mycelium, about 46 μg / g was produced. Table 1 shows the composition of the conjugated linoleic acid (CLA) of soybean fermented product of the edible mushroom mycelium.

Conjugated linolenic acid content
(μg / g)
Soybeans before fermentation
(Comparative Example 1)
Soybeans after fermentation Example 1
(Matsutake) Example 2
(Leaf mushroom) Example 3
(Mushroom crab) cis- 9, trans- 11 CLA nd 17.56 21.95 42.94 trans- 10, cis- 12 CLA nd 2.51 2.63 3.27 Total CLA nd 20.47 24.58 46.21 * All experiments were repeated three times
** nd; Not detected

Example  4 and 5:  Shrike mushroom  Production of powder through fermentation of bean solid using soybean

Yellow bean (product name: Daewon) was soaked for 12 hours and germinated at about 2 times of soaked soybeans in a dark room at 25 캜 to produce germinated soybean. The results are shown in Fig.

Sprouted beans were prepared by sprinkling two times as many grains while water was supplied to the bean sprouts in a 4 -

200 g of each of the soymilk beans (Example 4) and the sprouted beans (Example 5) were placed in a polystyrene fermentation vessel, and the mixture was incubated at 121 DEG C for 30 minutes, cooled to room temperature, ). The soybean solid fermentation was used for fermentation at 25 ℃ for 8 days.

Chungkookjang, which is produced by a typical solid fermentation method of soybean and widely used as food, was used as a control (Comparative Example 2). Chungkukjang was also soaked with yellow soybeans (foam name: Daewon bean) for 12 hours, and then the water was removed and treated at 121 ° C for 30 minutes. After cooling to room temperature, Bacillus inoculated 2TDJ15 amyloliquefaciens strain culture solution 5% of the total weight, which was produced by fermentation in 37 ℃ 3 days. Fermented soybean fermented product and Chungkukjang were dried at 50 캜 for 3 days and pulverized by a pulverizer to prepare a powder (Fig. 4).

Experimental Example  1. Fatty acids and Conjugated linoleic acid  content

The fatty acid and conjugated linolenic acid contents of the chungkukjang powder, the shiitake mushroom mycelia germinated fermented powder and the shodoshin mushroom fermented soybean powder prepared in Examples 4 and 5 were analyzed.

<Fatty Acid Analysis>

The fatty acid was carried out according to the fatty acid analysis method in the same manner as in Test Example 1 above.

The fatty acid content of the fermented soybean fermented product (Example 4) and the fermented product of germinated fermented soybean curd mushroom (Example 5) are shown in Table 2. Total fatty acid contents were as follows: Shiitake mushroom mycelium soaking fermented soybean powder> Chungkukjang powder> Magpie mushroom mycelium germination fermented soybean powder. Table 2 shows the comparison of fatty acid composition of soybean fermented powder of Chongkukjang and Shokuchishi mushroom mycelium.

Fatty acid content
(mg / 100 g)
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 5
(Sprouted beans) Saturated fatty acid   Palmitic acid (C16: 0) 2,529.7 3,449.0 1,985.0   Stearic acid (C18: 0) 540.4 1,072.2 605.9   Arachidic acid (C20: 0) 68.7 86.7 51.4   Behenic acid (C22: 0) 99.1 112.0 65.1   Lignoceric acid (C24: 0) 39.2 42.2 24.6 Total 3,277.1 5,025.0 2,882.2 Unsaturated fatty acid   Palmitoleic acid (C16: 1) 17.3 29.6 6.9   Oleic acid (C18: 1c) 3,602.9 4,893.1 2,756.1   Linoleic acid (C18: 2c) 5,252.8 6,435.5 3,302.3   α-Linolenic acid (C18: 3n3) 248.5 359.5 170.1   γ-Linolenic acid (C18: 3n6) 7.5 7.3 3.9   Eicosenic acid (C20: 1n9) nd 29.6 23.2   Eicosadienoic acid (C20: 2) 10.2 27.9 25.1 Eicosatrienoic acid (C20: 3n3) nd 3.5 2.2   Eicosatrienoic acid (C20: 3n3) 6.4 nd nd Arachidonic acid (C20: 4n6) nd 3.5 2.2   Nervonic acid (C24: 1n9) 4.7 23.3 13.7 Total 9,150.3 11,860.7 6,325.9 Total fatty acid 12,427.4 16,885.7 9,208.1 * All experiments were repeated three times
** nd; Not detected

&Lt; Analysis of conjugated linolenic acid &

The content of conjugated linolenic acid in the fermented soybean curd (Comparative Example 2), the fermented soybean curd submerged fermented soybean soybean (Example 4) and the fermented soybean fermented soybean curd subbed (Example 5) prepared in Examples 4 and 5, same. Conjugated linolenic acid was not detected in chungkukjang powder, and the total conjugated linolenic acid contents of fermented soybean curd flour and soybean curd fungus fermented powder were 55.22 μg / g and 50.68 μg / g, respectively. Table 3 shows a comparison of the content of conjugated linoleic acid in soybean fermented product powder of Chongkukjang and Shokuchishi mushroom mycelium.

Conjugated linolenic acid content
(μg / g)
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 4
(Sprouted beans) cis- 9, trans- 11 CLA nd 50.35 46.24 trans- 10, cis- 12 CLA nd 4.87 4.44 Total CLA nd 55.22 50.68 * All experiments were repeated three times
** nd; Not detected

Experimental Example  2. Total Phenolics  And isoflavone content

The fatty acid and conjugated linolenic acid contents of the chungkukjang powder, the shag mushroom soymilk fermented soybean powder and the shiitake mushroom fermented soybean powder prepared in Examples 4 and 5 were analyzed.

<Preparation of extract>

10 g of fermented soybean powder, 10 g of fermented soybean fermented soybean powder, and 10 g of 50% methanol were added thereto, followed by extraction at 300 rpm at room temperature for 12 hours. The extract was centrifuged for 20 minutes and then the supernatant was filtered with a 0.45 μm filter to prepare an extract. The extract was used for the analysis of total phenolics and isoflavones. The remainder was concentrated and dried at 0.5 mg / ml and 1.0 mg / ml Were used for antioxidant activity and digestive enzyme inhibitory activity.

&Lt; Analysis of total phenolics >

Total phenolic content was measured by Folin and Denis (1912). 0.5 mL of each extract was placed in a test tube, and 25% Na ₂ CO ₃ And then incubated for 3 minutes. Then, 0.25 mL of 2 N-Folin-Ciocalteu phenol was added and mixed. The mixture was developed at 30 ° C for 1 hour, and the absorbance at 750 nm was measured. Using gallic acid, (GAE mg / g) from the curve, and the results are shown in Table 4.

The content of chungkukjang powder (Comparative Example 2) was 11.67 mg / g, which was lower than that of soybean fermented soybean curd mushroom. Total phenolic contents of the fermented soybean fermented soybean powder and the fermented soybean fermented soybean powder were 12.07 mg / g and 13.05 mg / g, respectively. Table 4 shows the total phenolic content of soybean fermented powder of Chungkukjang and Shokuchishi mushroom mycelium.

Total phenolic content
(mg / g)
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 5
(Sprouted beans) Total phenolics 11.67 12.07 13.05 * All experiments were repeated three times

&Lt; Isoflavone analysis >

Isoflavone analysis And analyzed by HPLC according to the method of Cho et al. (2011). The analytical column was analyzed using a Lichrophore 100 RP C18 column (4.6 mm, 5 μm, Merck, Germany) and mobile phase solvent was analyzed with 0.2% acetic acid in water (mobile phase A) and 0.2% acetic acid in water (mobile phase B). The mobile phase conditions were 0 min -100%, 15 min-90%, 25 min -80%, 35 min -75%, 45 min-65% and 50 min-65% on the basis of solvent A. Each sample was injected with 20 μL of each extract and the mobile phase was maintained at 30 ° C at 1 mL / min. The detector was a diode array detector (Agilent 1200 series, Agilent Co.) and quantified at UV 254 nm.

The total isoflavone contents of the fermented powder of Chongkukjang powder, Shoku mushroom mycelia, and soaked fermented soybean powder were 1,583.88 ㎍ / g and 1,681.8 ㎍ / g and 1,418.84 ㎍ / g, respectively. The fermented product had much higher unglycosylated - isoflavone content. Particularly, in the case of the fermented germinating soybean mycelia (Example 5), the ratio of glycoside to isoflavone was 20.1% and that of non-glycoside to isoflavone was 79.9% Non-glycoside-isoflavone ratio of 14.8%) (Table 4 and Figure 5). Table 5 shows the isoflavone content of soybean fermented product powder of Chungkukjang and Shokuchishi mushroom mycelium.

Isoflavone content (μg / g) /
ratio (%)
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 5
(Sprouted beans) Glycosides - isoflavones   Daidzin 406.09 / 25.6 149.41 / 8.9 87.40 / 6.2   Glycitin 341.50 / 21.6 142.29 / 8.4 36.10 / 2.5   Genistin 602.20 / 38.0 266.64 / 15.9 162.28 / 11.4   Total 1,349.78 / 85.2 558.34 / 33.2 285.78 / 20.1 Non-glycoside-isoflavone   Daidzein 99.79 / 6.3 578.14 / 34.4 584.24 / 41.2   Glycitein 68.24 / 4.3 191.67 / 11.4 178.34 / 12.6   Genistein 66.39 / 4.2 353.66 / 21.0 370.47 / 26.1   Total 234.41 / 14.8 1,123.46 / 66.8 1,133.06 / 79.9 Total isoflavones 1,583.88 / 100 1,681.80 / 100 1,418.84 / 100 * All experiments were repeated three times
** nd; Not detected

Experimental Example  3. Free amino acid  And ammonia content

Free amino acid and ammonia contents of the chungkukjang powder, the shiitake mushroom mycelia germinated fermented powder and the shodoshin mushroom mycelial fermented soybean powder prepared in Examples 4 and 5 were analyzed.

&Lt; Free Amino Acid and Ammonia Analysis >

For analysis of free amino acid and ammonia, 0.1 g of powder was taken into a test tube according to the Food Ordinance method, 5 mL of HPLC water was added, and hydrolysis was carried out at 60 ° C. for 1 hour. 1 mL of 10% salicylic acid was added to each sample to obtain 2 The protein was allowed to settle for a time. The filtrate was filtered with a 0.2 μm membrane filter (Dismic-25CS, Toyoroshikaisha Ltd., Tokyo, Japan). The filtrate was dissolved in 2 ml of lithium citrate buffer (pH 2.2) The results were analyzed using an automatic analyzer, and the results are shown in Table 5 and FIG.

The total amino acid contents of chungkukjang powder, shrike mushroom mycelia sole fermented water powder and shawn mushroom mycelia germinated fermented powder were 1,580.29mg / 100g, 1,580.5mg / 100g and 1,886mg / 100g, respectively.

In the case of protein fermented foods, fermented soybean fermented soybean fermented soybean (25.52 mg / 100 g and 26.17 mg / 100 g) was the main factor of the occurrence of odor in the case of ammonia content than that of chungkukjang powder (ammonia content: 112.81 mg / ), Which is much lower than that of Chungkukjang powder. Table 6 shows the amino acid and ammonia content of soybean fermented powder of Chungkukjang and Shark mushroom mycelium.

Amino acid content
(Mg / 100 g)
Cheonggukjang
(Comparative Example 2)
Fermented soybean curd mycelium Example 4
(Soymilk) Example 5
(Sprouted beans) Non-essential amino acids   Proline nd 20.60 43.46   Aspartic acid 87.57 118.18 161.09   Serine 46.76 42.78 78.59   Glutamic acid 196.37 315.24 552.10   Sarcosine 44.43 9.32 8.85   Aminoadipic acid 78.73 20.27 17.07   Glycine 60.23 36.25 40.71   Alanine 10.44 6.11 57.11   Citrulline 27.33 5.59 8.88   alpha-aminobutyric acid 15.06 nd 8.54   Cystine nd 18.75 23.98   Tyrosine 26.06 37.18 20.61   β-domain 131.79 3.14 57.47   β-aminoisobutyric acid 177.63 nd 22.58   γ-aminobutyric acid 44.23 nd 22.94   Aminoethanol 104.91 7.00 1.82   Hydroxyproline 2.52 2.99 6.96   Ornithine 66.87 5.08 18.62   3-Methylhistidine nd nd 12.56   Anserine 65.08 21.40 28.87   Carnosine 8.47 3.28 9.39   Arginine 54.91 42.96 67.97   Totals 1,249.39 945.94 1,550.26 Essential amino acids   Threonine 33.17 17.14 38.32   Valine 38.85 35.80 44.25   Methionine 13.73 18.15 22.80   Isoleucine 32.42 16.03 34.57   Leucine 66.93 37.48 55.80   Phenylalanine 58.97 43.33 63.78   Lysine 70.81 33.25 58.08   Histidine 16.02 11.38 18.14   Total 330.9 212.56 335.74 Total amino acids 1,580.29 1,158.50 1,886.00 Ammonia 112.81 25.52 26.17 * All experiments were repeated three times
** nd; Not detected

Experimental Example  4. Antioxidant activity

OH 라디칼 소거활성으로 항산화 활성을 분석하였다. 표 7은 청국장과 까치버섯 균사체 콩발효물 분말의 항산화 활성 비교한 것을 나타낸다.The DPPH radical scavenging activity, the ABTS radical scavenging activity, and the antioxidative activity of the fermented powder of chrysanthemum powder, shiitake mushroom germination fermented product and shag mushroom mycelium prepared in Examples 4 and 5

The antioxidative activity was analyzed by OH radical scavenging activity. Table 7 shows the comparison of antioxidative activities of soybean fermented powder of Chongkukjang and Shoku mushroom mycelium.

Radical scavenging activity
(%)
Chungkookjang powder
(Comparative Example 4)
Mushroom mycelia soybean fermented water powder Example 2
(Soymilk) Example 3
(Sprouted beans) DPPH 52.49 58.74 62.49 ABTS 82.01 87.61 92.36

OH

OH 52.62 59.32 72.99 * All experiments were repeated three times
** Treatment concentration: 0.5 mg / ml

&Lt; DPPH radical scavenging activity >

The DPPH radical scavenging activity was measured according to the method of Blois (1958). 0.8 ml of 10 ^{- 4} mM DPPH solution and 0.2 ml of extract were added and reacted in a dark room for 30 minutes. Absorbance was measured at 525 nm using a spectrophotometer. In the negative control experiment, 0.2 mL of ethanol was used instead of the sample, and the DPPH radical scavenging activity was expressed as a relative value (%) of the absorbance of the experimental group and the negative control.

The fermented powder of soybean fermented mushroom mycelia showed higher activity than that of chungkukjang powder (Comparative Example 2), but germinated soybean showed more excellent activity. In other words, 52.49% of Chungkukjang powder and 62.49% of Shung-mushroom mycelia germinated fermented powder showed more DPPH radical scavenging activity than the 58.74% of Shung mushroom mycelium sole fermented powder (Table 7).

<ABTS radical scavenging activity>

The ABTS radicals (ABTS ⁺ ) were formed by mixing 7 mM ABTS solution and 2.45 mM potassium persulfate in 1: 1 (v / v) and left for 12 ~ 16 hours in the dark room. And then diluted with methanol to obtain an absorbance value of 0.7 at 732 nm. 0.9 mL of the appropriately diluted ABTS solution and 0.1 mL of the extract were mixed and the absorbance was measured at 732 nm using a spectrophotometer after 3 minutes. In the negative control experiment, 0.1 mL of methanol was used instead of the sample, and the ABTs radical scavenging activity was shown in the same manner as the DPPH radical scavenging activity described above.

The ABTS radical scavenging activity was also higher in the soybean fermented powder than in the chungkukjang powder (Comparative Example 2), but the germinated soybean showed more excellent activity (Table 7).

OH 라디칼 소거활성><

OH radical scavenging activity>

10 mM FeSO _4. 0.2 mL of 7H ₂ O-EDTA, 0.2 mL of 10 mM 2-deoxyribose, 0.2 mL of 10 mM H ₂ O ₂ , and 1.4 mL of a sample were mixed and reacted at 37 ° C. for 4 hours. 1 ml of 1% thiobarbituric acid and 2.8% trichloroaceric acid were added to the mixture, and the mixture was heated at 100 ° C for 20 minutes, cooled, and absorbance was measured at 520 nm. In the negative control experiment, PBS buffer solution (8.76 g NaCl, 0.11 g NaH ₂ PO ₄ , 0.596 g Na ₂ HPO ₄ ) was used instead of the sample. The hydroxyl radical scavenging activity was expressed as a percentage (%) of the absorbance difference between the addition of the sample solution and the addition of the sample solution (Adjimani and Asare, 2015).

OH 라디칼 소거활성 청국장 분말(비교예 2)은 52.62%로 까치버섯 균사체 콩발효물 분말이 활성이 낮았고, 까치버섯 균사체 발아콩발효물 분말이 72.99%로 가장 높은 활성을 나타내었다(표 7).

The activity of the radish scavenging active chungkukjang powder (Comparative Example 2) was 52.62%, the activity of the soybean fermented powder of the magpie mushroom was low, and the fermented powder of the germinated fermented soybean curd mushroom showed the highest activity of 72.99% (Table 7).

Experimental Example  5. Digestive enzyme inhibitory activity

The alpha-glucosidase inhibitory activity and the pancreatic lipase inhibitory activity of the fermented powder of chongkukjang powder, shiitake mushroom mycelia germinated soybean fermented product and shrike mushroom mycelial sole prepared in Examples 4 and 5, and carbohydrate and lipid digestive enzyme inhibitory activity Respectively. Table 8 shows the digestive enzyme inhibitory activity of soybean fermented powder of Chongkukjang and Shokuchishi mycelia.

Digestive enzyme inhibitory activity
(μg / g)
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 5
(Sprouted beans) Alpha-glucosidase 85.49 92.64 94.55 Pancreatic lipase 26.96 42.12 46.77 * All experiments were repeated three times
** Treatment concentration: 1.0 mg / ml

&Lt; Alpha-Glucosidase Inhibitory Activity >

To each 30 μL of each extract, 70 μL of 0.5 U / mL alpha-glucosidase enzyme solution was added, and 50 μL of 200 mM sodium phosphate buffer (pH 6.8) was mixed and preliminarily incubated at 37 ° C. for 10 minutes. Then, p -NPG ( p- nitrophenyl butyrate was dissolved in 100 μL of sodium phosphate buffer solution (pH 6.8), and the mixture was reacted at 37 ° C. for 10 minutes. Then, 750 mL of 100 mM Na ₂ CO ₃ was added to the reaction solution to stop the reaction and the absorbance at 420 nm was measured. Negative control experiments were conducted with extraction solvents instead of samples.

The activity of soybean fermented powder of Cryptomeria japonica was higher than that of Chungkookjang powder (Comparative Example 2), and germinated soybean showed the highest activity. In other words, 85.49% of Chungkukjang powder and 94.55% of Shung-mushroom mycelia germinated fermented powder showed 92% higher DPPH radical scavenging activity than 92.64% of Shiitake mushroom mycelium sole fermented powder (Table 8).

<Pancreatic Lipase Inhibitory Activity>

50 μL of each extract and 50 μL of 0.5 U / mL lipase enzyme were mixed with 50 μL of 200 mM sodium phosphate buffer, preliminarily cultured at 37 ° C. for 10 minutes, 100 μL of sodium phosphate buffer dissolved in p -NPG to 10 mM, The reaction was carried out for 10 minutes. The reaction was stopped by adding 750 mL of 100 mM Na ₂ CO ₃ to the reaction solution, and the absorbance was measured at 420 nm. Negative control experiments were conducted with extraction solvents instead of samples.

The pancreatic lipase inhibitory activity was also higher in the soybean fermented powder of the black mushroom mycelium than in the chungkukjang powder (Comparative Example 2), but the germinated soybean showed more excellent activity (Table 8).

Experimental Example  6. Evaluation of palatability

The palatability evaluation of the cheonggukjang powder, the shiitake mushroom mycelia germination fermented product powder and the shodoshin mushroom mycelia sole fermented product powder prepared in Examples 4 and 5 was firstly recognized for the purpose of palatability evaluation and samples, The results of this study were as follows: First, three excellent Stallmario food staffs and 20 students from Kyungnam University of Science and Technology were selected as excellent 5, good 4, medium 3, poor 2 and very good Bad 1 point '.

The results of the palatability evaluation are shown in Table 9, and the color was almost similar, but the fermented powder of germinated soybean curd mushroom mycelium was superior to that of chongkukjang powder in terms of odor, flavor, taste and taste. Especially, in the case of odor, the ammonia odor characteristic of protein fermented food was more severe than that of soaked mushroom mycelia sole fermented powder. Table 9 shows a comparison of the palatability of soybean fermented product powder of Chongkukjang and Shoku mushroom mycelium.

Purity evaluation item
Chungkookjang powder
(Comparative Example 2)
Mushroom mycelia soybean fermented water powder Example 4
(Soymilk) Example 5
(Sprouted beans) Color 3.98 4.02 4.06 stink 4.44 3.72 3.88 incense 3.71 4.43 4.32 flavor 4.09 4.23 4.31 Likelihood 4.01 4.22 4.18 * All experiments were repeated three times
** Treatment concentration: 1.0 mg / ml

Although specific portions of the present invention have been described in detail, those skilled in the art will appreciate that these specific embodiments are merely examples of preferred embodiments and that the scope of the present invention is not limited thereto. It is therefore intended that the scope of the present invention be defined by the appended claims and their equivalents.

Claims (16)

At least one of at least one of mushroom mycelium and culture thereof selected from magpie mushroom, pine mushroom, and mushroom; And
Fermented soybean fermented solid by mushroom, including soybeans added.
The method according to claim 1,
A fermented soybean fermented by mushroom solid fermentation, wherein at least one of the contents of the conjugated linoleic acid and the non-glycoside isoflavone is increased.
The method according to claim 1,
Fermented soybean fermented solid by mushroom, with reduced ammonia content and improved flavor.
The method according to claim 1,
The soybean is a fermented soybean which is solid-fermented by mushroom, which is soaked bean or germinated soybean.
A food comprising the soybean fermented product according to any one of claims 1 to 4.
6. The method of claim 5,
Wherein the food is processed into a form selected from a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, a tablet, a capsule and a liquid.
6. The method of claim 5,
Antioxidant activity, diabetes, or obesity.
An inoculation step of inoculating the soybean to which at least one of mushroom mycelia selected from mushroom mushroom, mushroom mushroom, and mushroom and at least one of the cultures thereof is added; And
And a fermentation step of fermenting the soybean.
An inoculation step of inoculating the soybean to which at least one of mushroom mycelia selected from mushroom mushroom, mushroom mushroom, and mushroom and at least one of the cultures thereof is added; And
And a fermentation step of fermenting the soybean.
10. The method according to claim 8 or 9,
Further comprising soaking or germinating the soybeans before the soybean is grown in said seeding step.
10. The method according to claim 8 or 9,
Wherein the mushroom mycelium or the culture thereof is inoculated in an amount of 5 to 10 parts by weight based on the weight of the soybean.
10. The method according to claim 8 or 9,
Further comprising culturing the mushroom at room temperature for 4 to 6 days prior to the inoculating step.
10. The method according to claim 8 or 9,
Wherein the fermentation step is performed at 20 to 30 DEG C for 5 to 10 days.
10. The method according to claim 8 or 9,
Further comprising drying and pulverizing after the fermentation step.
15. The method of claim 14,
Wherein the pulverizing comprises drying the fermented soybeans at 45 to 55 DEG C for 2 to 3 days.
10. The method according to claim 8 or 9,
The step of formulating the soybean fermented product into a powder, a granule, a flake, a paste, a syrup, a gel, a jelly, a bar, a pill, a tablet, a capsule and a liquid after the fermentation step &Lt; / RTI &gt;

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