KR20190138066A - kombucha manufacturing method - Google Patents

Abstract

Disclosed is a method of manufacturing sugar-excluded Kombucha, which facilitates drinking of Kombucha and increases anticancer and antioxidant effects by mixing one or more extracts among tangerine, redhyang, cheonhyehyang and hanrabong extracts, i.e., Jeju special products with a Kombucha base having a healthy sweet taste by replacing sugar with coconut sugar, i.e., a natural sweetener. The method according to the present invention is a method of manufacturing sugar-excluded Kombucha, comprising the steps of: preparing a Kombucha base; and mixing one or more of tangerine, redhyang, cheonhyehyang and hanrabong extracts with the Kombucha base. The step of preparing the Kombucha base may comprise: a step of injecting a coconut sugar powder into boiled spring water and stirring the coconut sugar powder with the boiled spring water until the coconut sugar powder is completely dissolved into the boiled spring water to prepare sugar water; a step of dipping a black tea bag and a green tea bag into the sugar water; a step of cold fermenting the sugar water having the tea bags dipped thereinto at a room temperature of 18 to 30°C for 10 days; a tea bag removing step of removing the tea bags from the sugar water; a step of mixing a fermentation promoting vinegar with the tea bag-removed sugar water; a step of injecting fermented Kombucha into the sugar water; a step of putting the Kombucha-injected sugar water into a container, sealing the container, and culturing the Kombucha-injected sugar water in the container for 7 to 20 days to prepare the Kombucha base; and a step of filtering the Kombucha base to remove impurities included in the Kombucha base.

Description

Kombucha manufacturing method without sugar {kombucha manufacturing method}

The present invention relates to a method for manufacturing kombucha, and more particularly, to Kombucha base, which has a healthy sweet taste by substituting a natural sweetener instead of sugar, one of Jeju's specialties such as tangerine, red flavor, Cheonhye flavor, Hallabong extract The present invention relates to a method for producing sugar-free comb tea, which is easy to drink by mixing the above extracts and has enhanced anticancer and antioxidant effects.

Kombucha is a fermented beverage obtained by fermenting by symbiotic mycelium of bacteria and yeast known as black tea mushroom in Korea. Kombucha originated from the ancient Qin dynasty of ancient China, and was called "sacred tea" because of its high detoxification and tonic effect.Then it was passed to Korea through Japan around 414 AD and is now mainly called "Tea Kvass." It's called "a lot of drinking".

Many studies have been conducted on the effects of Kombucha on the physiological activity and public health, and representatively, various physiological activities such as antioxidant activity, antimicrobial activity, anticancer activity, wound healing effect, and liver function protection through active oxygen production inhibition Reported. Among the studies on the anticancer activity of Kombucha, Jayabalan et al measured anticancer activity of various solvent fractions of Kombucha extract showed strong anticancer activity in ethyl acetate fraction. Malonic acid and vitexin have been found to be active ingredients.

Srihari et al. Measured the anticancer activity of Kombucha using PC-3, a prostate cancer of humans, and found that m-RNA expression of interleukin8 and VEGF was significantly reduced among the target molecules. It was confirmed that the activities of matrix metalloproteinases-2 and -9, which are known to be significantly reduced.

However, despite the various physiological activities of Kombucha, its application and utilization in the industry is remarkably low. The main reason is that the fermentation of Kombucha is carried out through the symbiosis of microorganisms, so that the exact species and distribution of microorganisms used are established. Microbial safety and toxicity of fermentation products to normal cells. In particular, research on the cytotoxicity of Kombucha is inconsistent, and there is much controversy. For example, the Centers for Disease Control and Prevention (CDC) in the United States, in 1995, reported two clinical cases from Kombucha drinking in Iowa, USA. Although the exact cause has not been determined, the possibility of cytotoxicity due to low acidity of Kombucha has been suggested, and Srinivasan et al. Showed the clinical results of four patients who developed after drinking Kombucha. The potential for toxicity was suggested, but the direct cause of this is unknown.

Other studies have reported no cytotoxicity of Kombucha.Bhattacharya et al. Reported that Kombucha was fed with 2 mL / kg or 1% (v / v) of distilled water for 3 months in a rat experiment. Long-term toxicity, weight, etc. have been reported to be nontoxic. The US FDA has also confirmed that Kombucha is manufactured for GMP (Good Manufacturer's Practice) and is suitable for consumption by consumers and is non-toxic. Previously reported cytotoxicities also have high acidity, excessive intake and unsanitary production. It may be associated with such.

On the other hand, with respect to the manufacturing method of Kombucha, the Republic of Korea Patent Publication No. 2015-0124258 discloses a comb secondary fermentation composition. Although the present invention describes a composition having an anticancer effect by containing a green tea solution, a culture solution and a herbal medicine extract, it has a disadvantage that it is difficult to drink because the taste of the herbal medicine is added to the taste of Kombucha.

In addition, the Republic of Korea Patent No. 0482308 discloses a method for producing a green tea Kombucha drink. In the present invention, the black tea extract is mixed with green tea extract and sugar, and then described by the fermented Kombucha fermented beverage prepared by fermentation, but has the disadvantage that it is difficult to expect the efficacy of the conventional Kombucha.

1. Registered Patent Publication No. 10-0482308 (registered March 31, 2005) 2. Korean Patent Application Publication No. 10-2015-0124258 (published 5 November 2015) 3. Registered Patent Publication No. 10-1797813 (November 8, 2017 registration)

Vinod H Nargund et al, 2012, Seminars in oncology, 39 (5): 559-572 2. Sang Kwon Byun, 2006, Master Thesis, Graduate School, Yonsei University OS Frankfurt et al, 2003, Anti-cancer Drugs, 14 (7): 555-561 H Steller, 1995, Science, 267 (5203): 1445-1449 5.WS Simonet et al, 1997, Cell, 89 (2): 309-319 6.V Schreiber et al, 2006, Nature Reviews Molecular Cell Biology 7: 517-528 7.Z Jin et al, 2005, Cancer biology & therapy, 4 (2): 147-171 SN Farrow et al, 1996, Curr Opin Genet Dev, 6 (1): 45-49 9.OS Frankfurt et al, 2010, Febs Journal, 277 (16): 3437-3448

The technical problem to be achieved by the present invention is to drink by mixing one or more extracts of the Jeju special products citrus, red flavor, Cheonhye flavor, Hallabong extract to Kombucha base, which has a healthy sweet taste by substituting coconut sugar which is a natural sweetener instead of sugar. It is an object of the present invention to provide a method for preparing sugar-free Kombucha, which is easy and enhances anticancer and antioxidant effects.

The technical problem to be achieved in the present invention is not limited to the technical problem mentioned above, and other technical problems not mentioned above may be clearly understood by those skilled in the art from the following description. There will be.

In order to achieve the above technical problem, manufacturing a comb secondary base; The method of manufacturing a kombucha without sugar comprising the step of mixing any one or more of citrus fruits, red scent, Cheonhyehyang, Hallabong extract to the kombucha base, the step of preparing the kombucha base, powder in boiled spring water Preparing a sugar water by injecting a coconut sugar on the phase and stirring until completely dissolved; Immersing black tea and green tea bags in the sugar water; Cooling and fermenting the sugar water in the state where the tea bag is immersed at a room temperature of 18 to 30 ° C. for 10 days; A tea bag removing step of removing the tea bag from the sugar water; Mixing fermentation promoting vinegar with the sugar water from which tea bag is removed; Injecting the fermented Matsutake mushroom into the sugar water; Preparing the Kombucha base by sealing the sugar water, into which the pine mushroom is added, in a container and incubating for 7 to 30 days; Comprising the step of filtering the Kombucha base to remove impurities contained in the Kombucha base; Comprising, provides a method for manufacturing a Kombucha excluding sugar.

In an embodiment of the present invention, the mixing ratio of the comb secondary base and the extract may be 6: 4.

In an embodiment of the present invention, the pine mushroom, a pre-treatment step of washing, drying and cutting the pine mushroom in water; After the mixture of the Hwangchil-tree extract and the sugar source in the pine mushroom prepared in the pretreatment step and inoculated with the fermentation bacteria, the fermentation bacteria are inoculated and fermented into sugar water, and the Hwangchil-tree extract is extracted by using an extraction solvent. Water or methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1 , 3-butylene glycol, propylene glycol or a mixed solvent thereof.

In an embodiment of the present invention, the hwangchil wood extract may be mixed 5 to 20 parts by weight based on 100 parts by weight of red mushrooms.

According to an embodiment of the present invention, by using coconut sugar, which is a natural sweetener instead of sugar, it can give a healthy sweet taste, and the composition comprising citrus (red flavor, Cheonhyeyang, Hallabong) Kombucha fermentation broth as an active ingredient for normal cells Without toxicity, it induces apoptosis of bladder cancer cells, in particular non-invasive (superficial) bladder cancer cells, resulting in an excellent killing effect on early bladder cancer cells. Therefore, it can be usefully used as a pharmaceutical composition or food composition for the prevention or treatment of bladder cancer.

The effects of the present invention are not limited to the above-described effects, but should be understood to include all the effects deduced from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a flow chart illustrating a method for manufacturing a comb tea without sugar according to the present invention.

Hereinafter, a preferred embodiment of the present invention will be described in detail. In the following description of the present invention, if it is determined that the detailed description of the related known technology may obscure the gist of the present invention, the detailed description thereof will be omitted. Throughout the specification, when a part is said to "include" a certain component, it means that it may further include other components, not to exclude other components, unless otherwise stated.

The present invention pays attention to the problem that the conventional Kombucha (kombucha) using a white sugar as a sweetener is not good for health and has a low antioxidant activity, and drinking is burdensome as having a unique sour taste, and solved this problem, Instead of eliminating white sugar, Kombucha base is manufactured by replacing coconut sugar with healthy sweetness, and the antioxidant properties of Jeju specialty products such as tangerine, red flavor, Cheonhye flavor and Hallabong extract are mixed. The present invention was proposed after confirming that it can be increased and easy to drink.

In one embodiment of the present invention, as shown in Figure 1, the Kombucha (a) after preparing the Kombucha base (S10), (b) the tangerine, red flavor, Cheonhyeyang, Hallabong extract in the Kombucha base Disclosed is a method of mixing any one or more of them (S20). Here, it is preferable that the mixing ratio of a comb secondary base and an extract liquid is 6: 4.

The manufacturing process of the comb secondary base is as follows.

In one step Powdered coconut in boiled spring water Sugar  After input, coconut Sugar  Stir until completely dissolved to prepare sugar water (S11).

Conventional Kombucha has a unique sour taste and is not easy to drink due to the characteristics of fermented black tea. Therefore, it is preferable to add the sugar component in order to suppress the sour taste. At this time, the amount of the coconut sugar may be added in a ratio of 30 to 120 g per 1 L (liter) of spring water, and more preferably 50 to 100 g may be added.

In an embodiment of the present invention, the spring water is preferably used in the Jeju spring water containing a large amount of minerals.

In addition, in the embodiment of the present invention, coconut sugar is put in place of the existing white sugar is a natural sweetener made by collecting the flower juice contained in the buds of the coconut tree to give a healthy sweetness. These coconut sugars are rich in minerals such as potassium, magnesium, zinc and iron, which help your body metabolize, boosting immunity and affecting hormone production. In addition, it contains vitamins B ₁ , B ₂ , B ₃ , and B, which help metabolize carbohydrates, fats, and proteins to help with fatigue.

In 2 steps , remind Sugar In water Black tea  And Green Tea Tea Bag  Inject (S12).

Here, the input ratio between the tea tea bag and the tea tea bag is preferably 1: 3. When the amount of green tea is increased in the mixing ratio of green tea and black tea, the taste is softened to obtain a high quality fermented beverage.When the ratio of black tea tea bag is increased, the color of the fermentation broth becomes dark brown and the astringent taste becomes strong. Because of this, the above ratio is most suitable.

In the present embodiment, but using a tea bag to shorten the manufacturing time, black tea extract and green tea extract, or black tea powder and green tea powder may also be applied.

In 3 steps , remind Black Tea Tea Encyclopedia Green tea bag  Added Sugar water  Cool fermentation for 10 days at 18 ~ 30 ℃ room temperature (S13).

When the sugar is added during the fermentation process, there is a problem that the fermentation period takes a lot, but the present application can shorten the fermentation time since coconut sugar, which is a natural sweetener, is already added.

Coconut sugar has an acid production promoting effect in all cases. That is, a beverage fermented with sugar as a fermentable sugar has less acid production than fermented with coconut sugar and takes a long time for fermentation.

On the other hand, when using black tea and green tea extract without using black tea tea bag and green tea tea bag, adding coconut sugar to black tea and green tea extract also has an acid production promoting effect. When the addition ratio of coconut sugar in the sugar content ratio is increased from 20% by weight to 100% by weight, the titratable acidity of the fermentation broth increases. For example, when the coconut shoe is fermented by mixing at least 50% by weight, it takes about 12 days to reach an optimum acidity of about 1.2% (w / v), and the soft taste of the fermented beverage is smooth and high quality.

As another example, when 10% (w / v) of sugar is added to a mixed solution of green tea and black tea extract, the acidity of the fermentation broth is about 10% (w / v) after 12 days of fermentation. When 10% (w / v) of sugar was added to the mixture and 01% (v / v) of alcohol was added, the titratable acidity was as high as 22% (w / v) after 12 days. However, when 0.3% (v / v) or 0.5% (v / v) of alcohol is added, the degree of change in titratable acidity is similar to that of no alcohol.

Therefore, the addition of alcohol (0.1% (v / v)) enables the production of high organic acid during a short fermentation period, 10 ~ 15% (w / v) titration acidity showing the optimum taste of fermented beverage by fermentation for about 7 days To reach.

In addition, when coconut sugar is used as a sugar substitute and alcohol is added, that is, 0.1% (v / v) alcohol is added to the initial green tea and black tea extract, and 50% or 80% by weight of sugar is used as fermentable sugar. When added in place of sugar, the effect of promoting acid production is greatly improved. Particularly, when 10% (w / v) sugar is mixed and added by replacing 0.1% (v / v) alcohol and sugar with more than 80% honey, the titratable acidity is 15% (w) / v) or more, and can reach 15% (w / v) or less of titratable acidity which indicates the optimum taste of the fermented beverage by fermentation of about 5 days.

Therefore, the addition of sugars prepared by replacing at least 50% by weight of sugar with coconut sugar to 0.1% (v / v) alcohol and fermentable sugars in the initial green tea and black tea extracts not only shortens the fermentation period but also honey and green tea and It is an optimal condition that enables the production of high-quality functional drinks including black tea ingredients.

In 4 steps , remind Sugar In water  Remove the tea bags (S14).

The sugar water fermented by putting the tea bag into the shooter water is passed through a filtering network to extract only the fermentation broth except the tea bag.

In 5 steps , Sugar In water  Cucumber vinegar for fermentation is mixed (S15).

To prepare the cucumber vinegar described above,

5-1. First clean cucumbers and crush them, then put them in a juicer to juice.

The washed cucumber is preferably pulverized to a size of 1 ~ 6cm, when the cucumber is pulverized to less than 1cm to be pulverized more than necessary to obtain the same yield and activity as when pulverized to 1 ~ 6cm. It is uneconomical, and when pulverizing in excess of 6 cm size, the time for juice is long, it is not preferable because the juice can not be obtained sufficiently.

Grinding the cucumber to 15 to 25 parts by weight of the juice of the juice juice juice is preferably added 15 to 25 parts by weight of apple concentrate, 40 to 60 parts by weight of purified water to 10 to 20 Brix °, more preferably 14 ~ It is desirable to have a sweetness of 16 Brix °.

The addition of sugars to the above-mentioned sugar is to ferment the total acidity, which is a quality standard of fruit vinegar, to 40% or more through the following alcohol fermentation and acetic acid fermentation.

In addition, the sugar added to the cucumber juice is to use one or two or more of fruit concentrate, honey, coconut sugar, preferably to use apple concentrate in the fruit concentrate.

Cucumber juice added with the sugar is called cucumber juice.

5-2. In the juice of the added sugar cucumber sugar is added to the ratio of 1: 0.05 ~ 0.15 to the weight ratio, then mixed with alcohol fermentation.

Jumo used in the alcohol fermentation is inoculated with Saccharomyces cerevisiae in a ratio of 1: 0.01 to 0.1 by weight in cucumber juice sugar adjusted to a sugar concentration of 10 ~ 20 Brix ° with apple concentrate at 45-55 ℃ After that, it is preferable to culture.

The alcohol fermentation is preferably inoculated with a ratio of 1: 005 ~ 015 to weight of cucumber juice sugar culture incubated for 7-10 days in a 25-30 ℃ constant temperature incubator.

When inoculated with jujube in the ratio of 1: 0.05 ~ 0.15 to the weight of the cucumber juice and fermented below 25 ℃ can not be derived content of alcohol and sugar suitable for vinegar can be difficult to prepare vinegar smoothly, When the added cucumber is inoculated at a ratio of 1: 0.05 to 0.15 to weight ratio and fermented above 32 ° C., alcohol fermentation may not be performed smoothly due to high temperature, making it difficult to prepare vinegar.

In addition, when inoculated in the ratio of 1: 0.05 ~ 0.15 to the weight of the cucumber added to the sugar and fermented at less than 4 days at 25 ~ 32 ℃, the content of alcohol, sugar content for vinegar could not be derived It may be difficult to prepare smoothly, and if the fermentation is more than 10 days, the alcohol content may increase to some extent, but the sugar content does not change any more, which is uneconomical.

In addition, the inoculation of the sugar-added cucumbers in the ratio of 1: 0.05 to 0.15 compared to the weight is to inoculate the sugar-added cucumbers evenly to ensure that the alcohol fermentation proceeds smoothly. When the seedlings are inoculated in the ratio of less than 1: 0.05 to the weight of the added cucumber, the sugar-added cucumbers may not be evenly inoculated and alcohol fermentation may not proceed smoothly. When inoculated at a ratio of more than 1: 0.15 by weight, the sugar-added cucumber is inoculated more than necessary to be inoculated more economically and takes longer to ferment alcohol.

Cucumber juice sugar fermented in the same manner as the alcohol is called alcohol fermentation broth.

5-3. The alcohol fermented alcohol fermentation broth is sterilized for 5 to 20 minutes at 65 ~ 80 ℃. Sterilization as described above is to sterilize harmful bacteria without impairing the activity of the components of the alcoholic fermentation broth. When the alcoholic fermentation of the alcoholic fermentation broth is less than 5 minutes at 65 ° C, sterilization is not performed due to low temperature. In case of sterilization for over 20 minutes at 80 ° C., the activity of alcohol fermentation broth components is impaired due to the high temperature, which makes it difficult to carry out the following steps.

5-4. The sterilized alcohol fermentation broth is centrifuged and filtered.

The alcohol fermentation broth is centrifuged at 200 to 300 rpm, and filtered with a filtration membrane of 0.42 to 0.47 μm to remove foreign substances such as rinds. When the sterilized alcohol fermentation broth is centrifuged at less than 200rpm, foreign substances such as rinds are not easily precipitated, so it is difficult to remove foreign substances, and when centrifuged above 300rpm, it is uneconomical to centrifuge more than necessary. At this time, centrifuged and filtered with a filtration membrane is called a cucumber filtrate.

The alcohol fermentation broth from which foreign substances such as rinds are removed by the centrifugation and filtration membranes is called a cucumber filtrate.

5-5. The cucumber filtrate is inoculated with the seed for fermentation of acetic acid.

The seed is incubated by inoculating Acetobacter sp (acetic acid bacteria) in the cucumber filtrate. Acetobacter sp (acetic acid bacteria) inoculated in the cucumber filtrate at a ratio of 1: 0.01 to 0.1 by weight to 150 at 25 ~ 35 ℃ It is preferable to incubate shaking culture for 70-75 hours at ~ 300rpm.

The Acetobacter sp (acetic acid bacterium) has a bactericidal ability to prevent food decay by killing bacteria causing food poisoning, such as Escherichia coli or staphylococcus, and is usually used to make malt vinegar, apple vinegar, brewed vinegar, and alcohol vinegar.

The cucumber filtrate was inoculated with the prepared seedlings to ferment acetic acid. The cucumber filtrate was inoculated at a ratio of 1: 0.05 to 0.15 by weight and stirred at 25 to 35 ° C. at 150 to 300 rpm for 10 to 15 days. Acetic acid fermentation is performed to derive alcohol and sugar content suitable for vinegar. At this time, the fermentation temperature is preferably 25 ~ 35 ℃, more preferably acetic acid fermentation at a temperature of 28 ~ 32 ℃.

When the seedlings are inoculated at a ratio of 1: 0.05 to 0.15 to the weight of the cucumber filtrate and fermented at less than 25 ° C., acetic acid bacteria cannot grow smoothly, so that the content of alcohol and sugars suitable for vinegar cannot be derived, so that the vinegar is smooth It may be difficult to prepare, when the fermentation exceeds 35 ℃ acetic acid may not grow due to the high temperature may not be fermentation.

In addition, inoculate the cucumber filtrate to the seed and fermented at less than 10 days 150rpm, less than 10 days at 25 ~ 35 ℃ may not be able to produce a good amount of alcohol and sugar to fit the vinegar, 300rpm, Fermentation beyond 15 days is more economical than necessary.

In addition, when inoculating the cucumber filtrate in the ratio of less than 1: 0.05, the seed may not be evenly inoculated in the cucumber filtrate may not be able to proceed acetic acid fermentation smoothly, at a ratio exceeding 1: 0.15 When inoculated, the cucumber filtrate is inoculated more than necessary with seed, which is uneconomical and takes longer for acetic acid fermentation.

Cucumber vinegar produced by the above method is preferably made of fruit vinegar quality standards of less than 10% residual alcohol content, 55 ~ 60 Brix ° sugar and 56 ~ 60% acidity.

In 6 steps , Fermented Sugar In water  Hong These mushrooms  Inject (S16).

The red mushroom is added after going through the following process before adding it to sugar water.

A pretreatment step of washing, drying and cutting red mushrooms may comprise the step of fermenting a mixture of Hwangchil-tree extract and sugar source in the pretreated red mushrooms.

6-1. Pretreatment stage

The red mushroom is washed, dried and chopped in water. The water is fresh water containing no salt. The drying is not limited as long as it removes the moisture of the red mushroom, and as an example, any one of natural drying, hot air drying, vacuum drying, etc. may be used. If cut to size is not limited to this. The cutting size is about 3 to 6 cm in length.

Through the pretreatment process, first, foreign matters such as soil and worms may be removed from the surface of the red mushroom, and the quality of the red mushroom fermented food may be prevented from being degraded by the bacteria inhabiting the foreign substance in the subsequent process. .

6-2. Fermentation stage

The step of fermenting by inoculating the fermentation bacteria by mixing the Hilchi chile extract and the sugar source to the red mushroom prepared in the pretreatment step.

In the present invention, the deciduous tree (Dendropanax morbifera) is a plant belonging to the family Elderaceae, diabetes, hypertension, blood circulation disorders, liver function development, antioxidant, osteoporosis, periodontal disease, immunity strengthening, food poisoning prevention, neurostable, antibacterial, It is known to be effective in anti-inflammatory and anti-cancer.

In the present invention, the hwangchil wood extract refers to a material extracted by using an extraction solvent, the extracted liquid may be used in liquid form or filtered, concentrated and / or dried.

The extraction solvent is water or methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO) , 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, can be extracted at room temperature or warmed under the condition that the active ingredient of the extract is not destroyed or minimized. Since the extraction degree and loss degree of the active ingredient of the extract may vary depending on the extraction solvent to be extracted, an appropriate extraction solvent should be selected and used. The extraction method is not particularly limited, for example, cold extraction, ultrasonic extraction, and reflux. Cold extraction and the like.

In addition, the hwangchil wood extract may be obtained through a conventional purification process in addition to the extraction by the extraction solvent. For example, through various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). It can also be obtained through the obtained fraction.

Filtration is a process of removing the suspended solid particles from the extract, it may be used to filter the particles using cotton, nylon or the like, ultrafiltration, cryofiltration, centrifugal separation, etc., but is not limited thereto. Drying the filtrate includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, infrared drying, and the like. You can add

In the present invention, the Hilchi chinensis extract includes all the extracts and purified products obtained at each step of extraction, fractionation or purification, their dilutions, concentrates or dried products.

In addition, the hwangchil wood extract of the present invention can be extracted using the leaves, branches, berries or roots of the hwangchil tree, preferably extracted using the leaves and branches of the hwangchil tree, but is not limited thereto.

In the present invention, any one of the sugar members can be used as long as it can cause an osmotic pressure as a medium for extracting the active ingredients of red mushrooms and hwangchil wood. For example, sugar, starch syrup, honey or oligosaccharides may be used or mixtures thereof may be used.

In one specific embodiment, honey in the sugar source contains about 12 times the content of invert sugar (sugar) compared to sugar, so the osmotic pressure occurs faster, so in the case of fermented food prepared by adding honey to the red mushroom, added sugar Compared to the fermented food prepared by the useful ingredient has a significant increase effect.

In the present invention, the fermentation is to decompose organic materials using enzymes fermented to produce useful materials, or to significantly increase the content of useful materials.

In the present invention, the fermentation bacterium is not limited so long as it has a property of fermenting food and can be used safely. For example, aspergillus oryzae, aspergillus kawauchi, aspergillus niger, Bacillus subtilis, yeast, lactic acid bacteria, acetic acid bacteria, butyric acid bacteria ), Heochi bacteria (hiochi bacteria) and lactic acid bacteria, etc. can be used, it is preferable to use the sulfur bacteria.

In one specific embodiment, in the case of fermented food prepared by inoculating fermented bacteria, the contents of polyphenolic compounds, which are useful components of beta-glucan and β-chil extract, are useful components of red mushrooms. Increased.

In the present invention, when the red mushrooms fermented foods are prepared using the sugar source and Hwangchil-tree extract together, there is an effect of improving the taste and flavor of the red mushrooms fermented foods.

In one specific implementation, when preparing red mushrooms fermented food using sugar extract and Hwangchil-tree extract could provide a red mushroom fermented food with increased taste, flavor, shelf life and overall preference.

In the present invention, the extract of Hwangchil-tree may be mixed in an amount of 5 to 20 parts by weight, preferably 8 to 15 parts by weight, based on 100 parts by weight of red mushrooms. If the efficacy of the weak and exceeds 20 parts by weight, there is a problem that the efficacy of the increase according to the content is weak.

In the present invention, the sugar source may be 40 to 80 parts by weight, preferably 50 to 75 parts by weight, more preferably 55 to 68 parts by weight based on 100 parts by weight of the red mushrooms. If the weight of the sugar source is less than 40 parts by weight, enzymes, useful components and fragrances may not be sufficiently eluted during fermentation, and contamination of the decayed bacteria may occur, and if it exceeds 80 parts by weight, all of the yeast mushrooms are killed.

In the present invention, the fermentation may be performed for 5 to 15 days, preferably 7 to 12 days at a temperature of 30 to 50 ℃, preferably 35 to 45 ℃ fermentation temperature is less than 30 ℃ or fermentation time is 5 If less than one day, useful ingredients may remain in the red mushroom, it is uneconomical because the fermentation efficiency is almost the same if the fermentation temperature exceeds 50 ℃ or fermentation time exceeds 15 days.

Example  1: manufacture of Hwangchil wood extract

The leaves and branches of the Hwangchil trees collected in July-August in Jeollanam-do were selected and washed to completely remove foreign substances and impurities, and then the surface moisture was removed. 1kg of dry hwangchil wood was pulverized and powdered, 10L of 70% ethanol was added and stirred for 24 hours. The extract stock was filtered through Whatman, Advantes, No2 filter paper, and concentrated under reduced pressure and lyophilized to prepare an extract of Hwangchil-tree.

Example  2: coconut Sugawa  Redness Using Hwangchil Tree Extract Two mushrooms  Fermented Food Manufacturing

2-1. Hongi Mushroom Pretreatment Step

100 g of red mushrooms were washed, and then dried in a 1m × 1m dry tray to remove water. Then, the fungi was cut into 4 cm lengths in a longitudinal direction.

2-2. Fermentation stage

The red mushroom of 2-1 was put in a glass bottle, and 60 g of coconut sugar and 10 g of Hwangchil-tree extract of Example 1 were mixed, and then inoculated with H. erythrocytes and fermented at 30 to 35 ° C. for 8 days.

Example  3: coconut Sugar  Hong Hong Two mushrooms  Fermented Food Manufacturing

In the fermentation step of Example 2-2, except for the fermented by mixing 60g coconut sugar to 60g of red mushrooms, the rest of the fermented food was prepared in the same manner as in Example 2.

Comparative example  1: Hong with Sugar and Hwangchil Tree Extracts Two mushrooms  Fermented Food Manufacturing

In the fermentation step of Example 2-2, except for fermented by mixing the white sugar 60g and the white sugar 60g and Hwangchil wood extract 10g in the fermentation step, the rest of the fermented food was prepared in the same manner as in Example 2.

Comparative example  2: red with sugar Two mushrooms  Fermented Food Manufacturing

In the fermentation step of Example 2-2, except for fermentation by mixing 60g of white sugar to 100g of red mushrooms, the rest of the fermented food was prepared in the same manner as in Example 2.

Example  4: Hong Two mushrooms  Determination of Useful Components in Fermented Foods

4-1 Determination of the Content of Beta-Glucan

100 ml of red mushrooms fermented food prepared in Example 2, Example 3, Comparative Example 1 and Comparative Example 2 was concentrated under reduced pressure and lyophilized to powder. Then, 100 mg of each sample was added to a culture bottle, and 15 ml of hydrochloric acid solution was added. Then, beta-glucan was extracted by mixing with a vertex mixer every 15 minutes for 45 minutes in an incubator maintained at 30 ° C. Then, 10ml of distilled water was added to the culture bottle, and heated to a temperature of 100 ° C for 2 hours, and then cooled, and 10ml of 2N KOH was added thereto. 200mM sodium acetate (pH5.0) was added to the sample of each culture bottle, and it adjusted to the volume of 100 ml. Each mixture was centrifuged at 14,000 rpm for 10 minutes to obtain a supernatant. The supernatant was then exo-1,3-beta-glucanase (exo-1,3-β-glucanse, 20 U / ml) and beta-glucose. Incubated for 60 minutes at 40 ° C. with a mixture of Sidayes (β-gludosidase, 4U / ml) and then using a beta-glucan assay kit sold by Megazyme Int, wicklow, Ireland The content was analyzed.

To quantify alpha-glucan, 2 ml of 2M KOH was added to 100 mg of each powdered sample, followed by stirring for 20 minutes in an ice bath. 8 ml of 12 M sodium acetate (pH 38) was added to each culture bottle, and a mixture of 02 ml amylolucosidase (amyloglucosidase (1630 U / ml) and invertase (500 U / ml) was added thereto. The culture was incubated with intermittent well for 30 minutes in the incubator to maintain the temperature of. Each culture was centrifuged at 14,000 rpm for 10 minutes to separate the supernatant. 0.1 ml of the supernatant was transferred to a glass test tube, and 0.1 ml of 200 mM sodium acetate (pH 50) and 30 ml of GOPOD solution were mixed and incubated for an additional 20 minutes. Alpha-glucan content was analyzed using a beta-glucan assay kit. It was.

The total amount of glucan was calculated by subtracting the amount of alpha-glucan, and each treatment was analyzed by three iterations to obtain an average value. Red mushroom powder was used as a control. The results are shown in Table 1.

sample Beta-Glucan Content (%) Control 35 Example 2 50 Example 3 48 Comparative Example 1 37 Comparative Example 2 38

As shown in Table 1, the beta-glucan content was increased by about 13 times when fermented with fermented bacteria compared to the red mushrooms not fermented. In addition, the red mushroom fermented foods of Example 2 and Example 3 fermented by adding coconut sugar was confirmed that the beta-glucan content was increased compared to the red mushroom fermented foods of Comparative Examples 1 and 2 fermented with sugar. It was.

4-2. Total Polyphenol Content Determination

100 ml of red mushrooms fermented food prepared in Example 2, Example 3, Comparative Example 1 and Comparative Example 2 was concentrated under reduced pressure and lyophilized to powder. Then, 2 ml of sodium carbonate was added to the samples and 1 g of the yellow lacquer extract of Example 1, followed by addition of 100 µl of 50% Folin-Ciocalteu reagent. The absorbance was then measured at 720 nm using a spectrophotometer and the total polyphenol content of each extract was determined based on the calibration curve of gallic acid. The results are shown in Table 2.

sample Total Polyphenol Content (mg / g) Example 1 1.04 Example 2 8.14 Example 3 7.85 Comparative Example 1 4.39 Comparative Example 2 0.64

As shown in Table 2, the hongyi mushroom fermented food of Example 2 prepared by adding fermentation of the Hwangchil-tree extract increased about 8 times the total polyphenol content compared to the Hwangchil-tree extract of Example 1 not fermented.

In addition, the red mushroom fermented food of Example 2 fermented with the addition of coconut sugar was found to have a higher total polyphenol content than the red mushroom fermented food of Comparative Example 1 fermented with sugar.

Example 5 Sensory Evaluation of Red Mushrooms Fermented Foods

The red mushroom fermented foods of Example 2, Example 3, Comparative Example 1 and Comparative Example 2 are to be taken for one month (30 days) targeting 50 adult males and 50 adult females. In addition, the sensory evaluation was conducted after establishing stable criteria for taste. The sensory items were evaluated on a five-point scale based on a total of three types of taste, aroma, shelf life, and overall preference. The results are shown in Table 3.

division flavor incense Zhejiang Symbol Example 2 8.9 7.2 7.1 9.1 Example 3 6.9 5.8 6.2 8.8 Comparative Example 1 6.5 6.9 4.5 7.1 Comparative Example 2 5.6 5.4 4.2 6.5

* Scoring criteria: Very good 10 points, Good 8 points, Normal 6 points, Bad 4 points, Very bad 2 points

As shown in Table 3, the red mushroom fermented foods using the coconut sugar and Hwangchilchi extract of Example 2 received a high evaluation score in all sensory evaluation items. This is because the natural sugars of coconut sugar are involved in the growth of yeast to increase fermentation efficiency, and the pharmacological components of Hwangchil-tree are implemented.

In addition, the red mushroom fermented foods including the Hwangchil-tree extract of Example 2 and Comparative Example 1, the flavor and shelf life is improved compared to the red mushroom fermented foods that do not contain the Hwangchil-tree extract of Example 3 and Comparative Example 2.

Thus, during the fermentation step, the sugar source promotes the extraction of useful ingredients to increase the workability of the red mushrooms fermented food, the fermentation with coconut sugar improves the pharmacological components, taste and aroma of Hwangchil wood and red mushrooms, This brings an improved effect. In addition, the pharmacological component of the Hwangchil wood may be implemented fermented foods hongyi mushrooms effective in antioxidant activity, liver function improvement, osteoporosis and strengthening immunity in addition to the physiological activity effect of the red mushrooms.

With 7 steps Self fermented red These mushrooms  remind Sugar Water and  Sealed in a container together, incubated for 7-30 days Kombucha  To prepare the base (S17).

This resulted in a better fermented Kombucha base by injecting the fermented red mushrooms into a container with sugar water and fermenting them once again.

In step 8 , the kombucha base is filtered to remove the impurities contained in the kombucha base (S18). Accordingly, the comb secondary base is provided in a pure liquid state containing no foreign matter.

Meanwhile, the present invention mixes any one or more of citrus fruits, red flavor, Cheonhye flavor, Hallabong extract to the Kombucha base provided as described above. An embodiment thereof will be described in detail.

Sample processing

Citrus peel extract, for example, was mixed with the Kombucha base. Here, the citrus peel extract was heated for 10 minutes at a rate of adding 200 mL of distilled water to 10 g of citrus peel.

Statistical processing

This experiment was carried out independently repeated 5 times. Experimental results were calculated using the SPSS Version 180Package Program (IBM, New York, NY, USA) for statistical analysis, and the average and standard deviation of each experimental group, Duncan's multiple range at P <005 level after t-test and ANOVA analysis. The test was used to verify the significance between treatment groups.

Example  One

20 g of pellicle was added to 100 mL of Kombucha culture broth obtained through preculture, and 200 μl of citrus fruit (Namhae Sangsa, Jeju, Korea) extract was incubated at 30 ° C. for 10 days.

Comparative example  One

Kombucha was incubated for 10 days under 30 ° C conditions by adding 100 mL of culture medium and 20 g of pellicle obtained by stirring 90 g of sugar in 800 mL of Kombucha base.

Experimental Example  1: Cell Culture

The B16F10 mouse dermal malignant tumor cell line, 5637 human bladder cancer cell line, and the raw 2647 cell line, a mouse macrophage, were purchased from the Korea Cell Line Bank (KCLB, Seoul, Korea) and 10% FBS in RPMI-1640 (Sigma-Aldrich Co) medium. (Gibco Inc San Francisco, CA, USA) and 1% Penicillin (Gibco Inc) were added and cultured at 37 ° C. and 5% CO 2.

Experimental Example  2 : ORAC assay

The ORAC values of the general comb tea of Comparative Example 1 and the citrus comb tea of Example 1 were measured by applying the method of Ou et al. (14). Samples, standards and reagents used in this experiment were diluted in 75 mM phosphate buffer (pH 74) solution. AAPH (150 mM) was incubated at 37 ° C for 15 minutes immediately before the experiment, followed by mixing 25 μL of 100-fold diluted sample with 120 μL of fluorescein (40 nM) in a 96-well plate and measuring 25 μμL of AAPH (150 mM) for measurement. Was added. The measurement was performed using a fluorescence microplate reader (Spectra Max i3, Molecular Devices, Sunnyvale, Calif., USA) at 37 ° C for 1 hour at excitation at 485 nm and emission at 520 nm for 2 minutes. The measured values were shown by substituting the standard curve of trolox after calculating the area under curve (AUC) of each sample.

The antioxidant powers of the Kombucha of Comparative Example 1 and the Kombucha containing the citrus extract of Example 1 were compared. Antioxidant activity of each sample was compared by measuring oxygen radical absorbance capacity (ORAC), and as a result, Kombucha is known to be excellent in protecting cells from free radicals due to its high antioxidant capacity. Bhattacharya et al. (12) measured antioxidative activity after feeding Kombucha in an experiment using diabetic model mice, and reported that they significantly inhibited the reduction of glutathione in hepatocytes of mice fed Kombucha compared to the control group fed unfermented tea. I've done it. The citrus extract-containing Kombucha prepared in this study was found to increase about three times as much as the antioxidant power of normal fermented Kombucha. This increase in antioxidant power indicates that there is a strong antioxidant in citrus peel. It can be said. In fact, the antioxidant effects of citrus peel have been reported in many previous studies.

Although the above-described embodiment has been described with respect to the preferred embodiment of the present invention, it is noted that the present invention is not limited thereto and may be implemented in various forms without departing from the technical spirit of the present invention.

Claims (4)

Preparing a comb secondary base;
In the method of manufacturing a Kombucha excluding sugar consisting of mixing any one or more of citrus, red, Cheonhyeyang, Hallabong extract to the Kombucha base,
The step of preparing the Kombucha base, a step of preparing a sugar water by adding a powdered sugar sugar to the boiled spring water and then stirred until completely dissolved; Immersing black tea and green tea bags in the sugar water; Cooling and fermenting the sugar water in the state where the tea bag is immersed at a room temperature of 18 to 30 ° C. for 10 days; A tea bag removing step of removing the tea bag from the sugar water; Mixing fermentation promoting vinegar with the sugar water from which tea bag is removed; Adding fermented red mushrooms to the sugar water; Preparing a kombucha base by culturing for 7-30 days by putting the sugar water into which the red mushroom was added and putting it in a container; Filtering the Kombucha base to remove impurities contained in the Kombucha base; consisting of, sugar-free combcha manufacturing method.
The method according to claim 1,
The mixing ratio of the Kombucha base and the extract is 6: 4, wherein the method of manufacturing a sugar-free Kombucha.
The method according to claim 1,
The red mushrooms,
A pretreatment step of washing, drying and rinsing the red mushroom in water;
After the mixture of Hwangchil-tree extract and the sugar source in the red mushroom prepared in the pretreatment step and inoculated with the fermentation bacteria, the fermented bacteria were inoculated and added to sugar water,
The hwangchil wood extract is extracted using an extraction solvent, the extraction solvent is water or methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, the method for producing a sugar-free Kombucha.
The method according to claim 3,
The Hwangchil-tree extract is a comb tea production method is excluded sugar, characterized in that 5 to 20 parts by weight based on 100 parts by weight of red mushrooms.

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