KR101381950B1 - Fermented tea of persimmon leaf and manufacturing method of the same - Google Patents

Abstract

The present invention relates to a manufacturing method of persimmon leaf fermented tea which comprises: a step of pretreating persimmon leaves; a step of inoculating and fermenting monascus sp. or aspergillus sp. into the pre-treated persimmon leaves; and a step of drying the fermented persimmon leaves. Persimmon leaf fermented tea according to the present invention has various functionality of such cholesterol-lowering activity or antioxidative activity because useful biologically active substances such as monacolin K, polyphenols, flavonoids increase by microbial fermentation. Therefore, in case of drinking the persimmon leaf fermented tea according to the present invention by brewing it with hot water, etc., it is good for maintaining or improving health of a taster.

Description

TECHNICAL FIELD The present invention relates to a fermented tea of persimmon leaves and a manufacturing method thereof,

The present invention relates to a persimmon leaf fermented tea, and more particularly, to a persimmon leaf fermented tea improved in various functions such as cholesterol lowering activity or antioxidant activity by fermentation of microorganisms and a method for producing the same.

The persimmon is a special product of oriental origin, which is produced mainly in Korea, China and Japan and has a unique sweetness. Approximately 400 species of persimmon are distributed throughout the country as deciduous broad-leaved arboreous trees. Persimmon leaves have long been used in the private sector as a source of persimmon leaves. The clinical pharmacokinetics and efficacy of persimmon leaves are well documented in various documents such as Dongbu Bohak and Bon - gok Gangmok. Studies on the components and effects of persimmon leaves have recently been made. Persimmon leaves are diseases such as hypertension, arteriosclerosis, heart disease, and gastric ulcers. Duodenal ulcer, diabetes and other chronic diseases are known to be effective, and is also known to have an effect on cancer prevention. It is also known that flavonoids isolated from persimmon leaves inhibit the proliferation of tumor cells. Specifically, Persimmon Leaves are flavonoid glycosides such as astragalin, myricitrin, tannins, polyphenols, resins, coumarin-like compounds, betulic acid, oleanolic acid, And ursolic acid, chlorophyll, and vitamins A and C. The content of vitamin B1, pantothenic acid and folic acid is high, and it is recommended as a good food for prevention of adult diseases.

It was reported that DDPH radical scavenging activity, lipid peroxidation inhibitory effect and α-amylase inhibitory activity were increased in proportion to the content of polyphenols contained in persimmon leaves [Hong Jeonghui et al. (2008): Dried persimmon, (1999): Salmonella typhimurium strain TA98, 100, the extracts of persimmon leaf tea, green tea, oolong tea were found to be effective in inhibiting the mutation Mutagenic Effects of Korean Nutritional Supplements, Korean Journal of Food Science and Nutrition. Korean Patent Laid-Open Publication No. 10-2003-0024111 discloses a composition for lowering blood lipid concentration including persimmon leaf extract. Korean Patent Laid-Open Publication No. 10-2010-0055952 discloses a composition for reducing Helicobacter pylori An antimicrobial extract is disclosed. However, the above prior arts focus on using a physiologically active substance present in the persimmon leaf itself by a method such as extraction, and do not disclose a method for improving the functionality of persimmon leaf. In general, despite the presence of various physiologically active substances in persimmon leaves, there is no remarkable difference with other plants, and therefore, there is not much developed medicinal materials using them. In order to overcome this, it is important to increase the content of the physiologically active substance of the persimmon leaf by further treating the persimmon leaf rather than using the physiologically active substance present in the persimmon leaf itself. In this regard, Korean Patent Laid-Open Publication No. 10-2001-0103256 discloses that Monascus cream ( Monascus) ruber or Monascus purpureus inoculated and cultured to suggest the possibility of producing a persimmon leaf tea having a function such as blood pressure lowering and cholesterol lowering. However, the above-mentioned prior art does not have a specific embodiment for the manufacture of persimmon leaves, and the functional properties of fermented persimmon leaves have not been actually verified, and thus it is unclear whether the persimmon leaves can be used as functional materials.

SUMMARY OF THE INVENTION The present invention has been made under these circumstances, and an object of the present invention is to provide a persulfate fermented tea having various functionalities such as cholesterol-lowering activity or antioxidative activity, etc., which are enhanced by an increase in useful physiologically active substances such as monacolin K, polyphenol or flavonoid, And a method of manufacturing the same.

The inventors of the present invention have found that when persimmon leaves are fermented by specific microorganisms in the process of fermenting persimmon leaves with various microorganisms to produce fermented tea, useful physiologically active substances such as monacolin K, polyphenol or flavonoid are increased and cholesterol lowering Activity or antioxidative activity, and the present invention has been completed.

In order to solve the above-mentioned problem, the present invention provides a method for producing a fermented soybean oil, comprising the steps of: The pre-treated persimmon leaves were seeded with Monascus husk pilosus culture or Aspergillus tereus culture; And drying the fermented persimmon leaves.

In addition, the present invention relates to a process for producing The present invention also provides a persimmon leaf fermented tea characterized by being produced by fermenting Persimmon leaf with Aspergillus tereus or Aspergillus tereus .

The fermented tea of the persimmon leaves according to the present invention may exhibit various functionalities such as cholesterol lowering activity or antioxidative activity by increasing useful physiologically active substances such as monacolin K, polyphenol or flavonoid by microbial fermentation. Therefore, when the fermented tea of the persimmon leaves according to the present invention is drunk with hot water or the like, it is advantageous to maintain or improve the health of the tasting person.

Fig. 1 shows dry weight changes of P. gonorrhoeae cells contained in the culture broth after 10 days of incubation at 28 ° C in a liquid medium such as Mizutani medium, PDB medium, S-INAE medium and Kumhwa medium Graph.
FIG. 2 is a photograph of the persimmon fermented tea prepared in Production Example 6, and FIG. 3 is a photograph of the persimmon leaf tea produced in Comparative Production Example 1. 2, the persimmon fermented tea was obtained when the fermentation time was 6 days.
4 shows the HPLC analysis peak of the persimmon fermented tea obtained when the fermentation time was 6 days in Production Example 6, and Fig. 5 shows the HPLC analysis peak of the persimmon leaf tea prepared in Comparative Production Example 1. Fig.
6 is a graph showing the content of monacolin K in the persimmon fermented tea prepared in Production Examples 9 to 11 by fermentation time.

Hereinafter, the present invention will be described in detail.

A method of manufacturing a persimmon leaf fermented tea according to the present invention comprises the steps of pretreating persimmon leaves, fermenting the pretreated persimmon leaves, and drying the fermented persimmon leaves . Hereinafter, a method of manufacturing a persimmon fermented tea according to the present invention will be described step by step.

Pre-treatment of persimmon leaves

In the process for preparing a persimmon leaf fermented tea according to the present invention, the step of pretreating the persimmon leaves may be carried out by changing the physical or chemical state of the persimmon leaves to increase the efficiency of the fermentation step described later, further sterilizing or sterilizing the fermentation raw material, . In the present invention, the step of pretreating the persimmon leaves comprises concretely growing the persimmon leaves or drying the persimmon leaves. In the present invention, steaming steam means steaming the persimmon leaves, and the steaming means adding liquid such as water or liquid medium to the persimmon leaves and blowing steam to sterilize the persimmon leaves. The above-mentioned method of increasing the temperature is not limited to a kind of atmospheric pressure, pressurization, etc. Specifically, a predetermined water or a liquid medium is added to the persimmon leaves and mixed, and then the mixture is heated to a predetermined temperature. When adding the persimmon leaves, the amount of water or the liquid medium is not limited. However, it is preferable that the amount of the water or the liquid medium is 50 to 150 ml per 100 parts by weight of the persimmon leaves, Considering the efficiency of the drying step, it is preferably 75 to 125 ml. In addition, the temperature for growing the persimmon leaves is preferably from 100 to 125 캜, more preferably from 110 to 125 캜. In addition, from the viewpoint of ensuring the state change and sterilization effect of the persimmon leaves, the time for the increase is preferably 5 to 60 minutes, more preferably 10 to 30 minutes. In the present invention, the pretreatment step may further include a step of adding a liquid medium or water to the dried persimmon leaf and sterilizing the persimmon leaf when the persimmon leaf is pretreated by steam-drying and drying. At this time, the sterilization can also be preferably performed by the above-described growth method. When the persimmon leaves pretreated by the above-mentioned steamed and dried processes are sterilized, the amount of the liquid medium or water to be added is preferably 50 to 150 ml per 100 g of the dried persimmon leaves. When considering the efficiency of sterilization and the efficiency of the drying step after fermentation of persimmon leaves, To 125 ml. In addition, when sterilized persimmon leaves pretreated by the above-mentioned wet-growing and drying, the sterilization temperature is preferably 110 to 125 ° C, and the sterilization time is preferably 10 to 30 minutes, more preferably 10 to 20 minutes. The liquid medium to be added to the persimmon leaf when sterilized with the persimmon leaf or sterilized by the above-mentioned steaming and drying is not limited as long as it is suitable for the microorganism used in the fermentation step of the persimmon leaf described later, , 3.0 g of red bean powder, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85-110 ml of distilled water (preferably 90-100 ml of distilled water) Lt; / RTI > medium (Kumhwa medium); 1.0 g of maltose, 6.0 g of maltose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water (preferably 90 to 100 ml of distilled water) (Kumhwa medium-modified); 1.0 g of red bean powder 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO ₄ and 85-110 ml of distilled water (preferably 90-100 ml of distilled water) (Kumhwa medium-optimized); Or a medium in which two or more of them are mixed.

The step of fermenting the pretreated persimmon leaves

The step of fermenting the persimmon leaves pretreated in the method of manufacturing a persimmon leaf fermented tea according to the present invention is for fermenting persimmon leaves with specific microorganisms to produce new active ingredients or to increase the amount of active ingredients contained in persimmon leaves themselves. The step of fermenting the persimmon leaves pretreated in the present invention comprises inoculating a pre-treated persimmon leaf into a monacasque genus or an Aspergillus strain and culturing a microorganism inoculated at a predetermined temperature for a predetermined period of time.

At this time, the strain of Monascus crassacea used as a seed bacterium is not limited in its kind as long as it belongs to the genus Monascus and is capable of producing monacoline K as a secondary metabolite. For example, Monacus crassillus strain Monascus pilosus , Monascus ruber), Pseudomonas coarse frame buffer mouse (Monascus purpureus , Monascus kaoliang , Monascus barykery ) and Monascus anka ( Monascus anka ). In particular, considering the improvement in the amount of monacolin K produced during fermentation of persimmon leaves, the total content of polyphenol compounds in fermented persimmon leaves, the content of total flavonoid compounds, or the antioxidant activity, the Monacaschus strain used as a seed strain is called Monacus spp. ( Monascus pilosus ). The above-mentioned Monascus pilosus may specifically be KCCM 60084, IFO 4480 or KCCM 60396, and the deposit number is IFO 4480 or KCCM 60396. In addition, the type of Aspergillus strain used as a seed bacterium is not limited as long as it is a bacterium capable of producing useful active ingredients by fermenting persimmon leaves. The amount of the monacolin K produced during fermentation of the persimmon leaves, Aspergillus tereus is preferable considering the improvement of phenol compound content, total flavonoid compound content or antioxidant activity. The type of Aspergillus tereus is not limited as long as it exhibits characteristics similar to those of the genus Monacus sp. In terms of the dominant product. For example, it is preferably ATCC 21542.

In addition, in the step of fermenting the pretreated persimmon leaves, the seed is preferably selected from the group consisting of Monascus pilosus ) culture medium or Aspergillus tereus culture medium. At this time, the above-mentioned Monascus < RTI ID = 0.0 > pilosus) culture or Aspergillus terephthalate mouse (Aspergillus tereus culture medium was Monascus < RTI ID = 0.0 > pilosus) strain or Aspergillus terephthalate mouse (Aspergillus tereus ) in a liquid medium. At this time, Monascus < RTI ID = 0.0 & gt ; pilosus) culture or Aspergillus terephthalate mouse (Aspergillus Pseudomonas coarse Philo suspension or Aspergillus strains (Monascus pilosus) Aspergillus present in tereus) culture's mouse terephthalate (Aspergillus tereus ) is generally present in the form of a mycelium. It is preferable that the mycelium is homogenized by physical stirring, mechanical cutting or the like in consideration of the aspect of uniformly using the persimmon leaves as a substrate. In addition, Monascus < RTI ID = 0.0 & gt ; pilosus) culture or Aspergillus terephthalate mouse (Aspergillus tereus ) culture medium is not limited to a wide range as long as it can increase the number of bacterial strains or improve the production ability of useful active ingredients such as monacolin K. For example, PDB potato dextrose broth) medium, Mizutani medium (5.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.2 g of potassium acetate, 0.1 g of NaCl, 100 ml of distilled water, Sodium chloride (2.0 g of rice flour, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.2 g of sodium nitrate, 0.1 g of NaCl, 100 ml of distilled water, pH 6.0) And the like. In addition, the liquid medium used in the production of the seed culture is Monascus < RTI ID = 0.0 & gt ; pilosus) strain or Aspergillus terephthalate mouse (Aspergillus Considering the shortening of the growth period or the growth of the tereus strain and the improvement of the ability of producing monacolin K, 3.0 g of red bean powder, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , MgSO ₄ .7H ₂ O 0.05 g of NaCl, 0.1 g of NaCl and 85 to 110 ml of distilled water (preferably 90 to 100 ml of distilled water) in a Kumhwa medium; 1.0 g of maltose, 6.0 g of maltose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water (preferably 90 to 100 ml of distilled water) (Kumhwa medium-modified); 1.0 g of red bean powder 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO ₄ and 85-110 ml of distilled water (preferably 90-100 ml of distilled water) (Kumhwa medium-optimized); Or a medium in which two or more of them are mixed. In addition, when producing the seed culture, Monascus pilosus) strain or Aspergillus terephthalate mouse (Aspergillus tereus strain is preferably activated on a potato dextrose agar medium (PDA medium).

In addition, the amount of inoculum of the seedlings is not limited in the step of fermenting the pretreated persimmon leaves, but it is preferably 1 to 40 ml per 100 g of dried persimmon leaves considering economical efficiency of the fermented tea of persimmon leaves and efficiency of the drying step after fermentation of persimmon leaves And more preferably 2 to 20 ml per 100 g of dried persimmon leaves. In addition, the fermentation temperature and fermentation time in the step of fermenting the pretreated persimmon leaves, that is, the culture temperature and incubation time of the strain may have various ranges depending on the kind of the used strain. For example, , While simultaneously maximizing the total polyphenol compound content, total flavonoid compound content, or antioxidant activity. Considering the above conditions, the fermentation temperature in the step of fermenting the pretreated persimmon leaves is preferably 25 to 35 ° C and preferably 26 to 30 ° C. The fermentation time is preferably 2 to 6 days, more preferably 2 to 4 days. If the fermentation time exceeds 4 days, the amount of monacolin K is increased but the total polyphenol compound content, total flavonoid compound content, or antioxidant activity may decrease.

Step of drying fermented persimmon leaves

In the method of manufacturing a persimmon leaf fermentation tea according to the present invention, the step of drying the persimmon leaf fermented by reducing the water content of the fermented persimmon leaf to a proper level or less, for example, 10% or less, preferably 5% or less, And to improve storage stability. The step of drying the persimmon leaves fermented in the present invention comprises treating the fermented persimmon leaves at a predetermined temperature for a predetermined time using a known dryer such as a hot air dryer. At this time, the drying temperature of the fermented persimmon leaves is not limited. However, considering the stability of the active ingredient generated or increased by fermentation, it is preferably 80 ° C or lower, and 45 ° C to 65 ° C is preferable considering economical efficiency of the drying step. The drying time of the fermented persimmon leaves may be selected from various ranges depending on the water content and drying temperature of the fermented persimmon leaves, and is generally selected within the range of 3 to 20 hours, preferably 4 to 15 hours.

Another aspect of the present invention relates to a persimmon leaf fermentation tea, wherein the persimmon leaf fermentation tea according to the present invention is preferably a persimmon leaf fermented by Monascus pilosus ) or Aspergillus tereus (fermented with Aspergillus tereus ). Also, the persimmon fermented tea according to the present invention can be manufactured by the above-described method for producing persimmon fermented tea, and useful physiologically active substances such as monacolin K, polyphenol or flavonoid are increased, and various kinds of substances such as cholesterol- Functionality can be demonstrated. For example, the content of monacolin K in the persimmon fermented tea according to the present invention may be 50 to 350 μg, preferably 100 to 350 μg, more preferably 150 to 300 μg . In addition, the content of total polyphenol compounds in the persimmon fermented tea according to the present invention may be 60 to 65 占 퐂 / g of persimmon leaf fermentation tea. The total flavonoid compound content of the persimmon fermented tea according to the present invention may be 275 to 285 g per g of the persimmon leaf fermentation tea. In addition, the radical scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl radical) assay of the persimmon fermented tea according to the present invention may be 92 to 97%.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are intended to clearly illustrate the technical features of the present invention, and do not limit the scope of protection of the present invention.

One. Hong Guk,  Screening of liquid medium to prepare culture

(1) Selection of a liquid medium considering the growth of P. gum

As a known liquid medium for culture of P. ginseng, PDB (potato dextrose broth) medium, Mizutani medium (5.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.2 g of potassium acetate (2.0 g of rice flour, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.2 g of sodium nitrate 0.2 g, 0.1 g of NaCl, 100 ml of distilled water, pH 6.0). However, such a liquid medium has a problem that the growth of P. gonorrhoeae is slow and it is not economically feasible to use it as a liquid medium for producing a seed culture. The inventors of the present invention investigated a liquid medium capable of enriching the growth of P. gonorrhoeae in a short period of time. Among them, 3.0 g of red bean powder, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 91.05 ml of distilled water was developed and named as Kumhwa medium.

Fig. 1 shows dry weight changes of P. gonorrhoeae cells contained in the culture broth after 10 days of incubation at 28 ° C in a liquid medium such as Mizutani medium, PDB medium, S-INAE medium and Kumhwa medium Graph. With honggukgyun inoculated in a liquid medium which is a kind of honggukgyun Monascus pilosus IFO 4480 (the same strain as KCCM 60396 strain) was cultivated in potato dextrose agar medium (PDA medium) for 7 days, and then activated with agar block (about 1 cm x 1 cm) Agar block) was used.

As shown in FIG. 1, the gold culture medium can reduce the growth period of P. gingko by about 2 to 6 days and the production amount of the seed culture can be increased by 90 to 180% or more on the 10th day of cultivation .

(2) Selection of liquid medium considering the amount of monocholin K produced by Hong Guk Kim

While it is trying to consider the angular amount of choline K honggukgyun study the liquid medium improves the gold medium (Kumhwa medium) previously selected, red bean powder 1.0g, 6.0g maltose, peptone _{2.0g, KH 2 PO 4 0.8g,} MgSO 4 · 7H ₂ O, 0.1 g of NaCl and 100 ml of distilled water was developed and named Kumhwa medium-modified medium. The gold coin and the modified coin coin were respectively labeled with Monascus pilosus IFO 4480 was inoculated and cultured at about 28 ° C for 4 days to obtain a culture of P. gracilis. The monacolin K present in the obtained cultures of P. aeruginosa was analyzed using high performance liquid chromatography (HPLC). Specifically, the culture of P. aeruginosa was filtered with a 0.45 mu m filter to obtain a filtrate. The filtrate was then filtered through a filtration apparatus (Lee et al., Lee, SI, Kim, JW, Lee, YK, Yang, SH, Lee, IA, Suh, JW and Kim, SD ) Anti-obesity effect of Monascus pilosus Mycelial Extract in high fat diet- induced obese rats. J. Appl . Biol . Chem . 54: 197-205), the content of monacolin K per liter of culture medium of P. gingkoii was measured. As a result, the content of monacolin K in the culture solution of Hongkuk culture obtained by using Kumhwa medium as a liquid medium was 25 mg / l, and the concentration of the monocholin K in the culture solution of Hongkuk culture obtained by using Kumhwa medium- Of monacolin K was 65 mg / l.

In addition, while studying the liquid medium in which the Kumhwa medium was improved in consideration of the growth of P. gonorrhoeae and the amount of monacolin K produced, 1.0 g of red bean powder, 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO ₄ , and 100 ml of distilled water. The liquid medium was designated Kumhwa medium-optimized medium. As an optimal gold coin, Monju pilosus IFO 4480 was inoculated and cultured at about 28 ° C for 10 days to obtain a culture of R. guti. The cell mass and the content of monacolin K of P. gingkoifera in the obtained culture solution of P. gingkoii were analyzed. Monacolin K was analyzed using high performance liquid chromatography (HPLC). Specifically, the culture of P. aeruginosa was filtered with a 0.45 mu m filter to obtain a filtrate. The filtrate was then filtered through a filtration apparatus (Lee et al., Lee, SI, Kim, JW, Lee, YK, Yang, SH, Lee, IA, Suh, JW and Kim, SD ) Anti-obesity effect of Monascus pilosus Mycelial Extract in high fat diet- induced obese rats. J. Appl . Biol . Chem . 54: 197-205), the content of monacolin K per liter of culture medium of P. gingkoii was measured. As a result, the cell mass and the monacolin K content in the culture medium of Hongkuk culture obtained using the Kumhwa medium-optimized medium as the liquid medium were 50 mg / ml and 30 mg / l, respectively.

2. Preparation of culture medium for seed culture

Production Example 1

Monascus purpureus The KCTC 6120 strain was activated for 7 days in a potato dextrose agar medium (PDA medium), and agar blocks containing about 1 cm x 1 cm of P. vaginalis were cut out. Thereafter, the cut agar block was added to 100 ml of an optimal gold culture medium (1.0 g of red bean powder, 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO ₄ , 100 ml) and cultured at about 28 DEG C for 10 days at a stirring speed of 150 rpm to prepare a culture broth of P. gingkoii. Then, the culture of P. gracilis was stirred with a mixer to homogenize the mycelia of P. japonica and obtain a culture of Monascus purpureus for the bacterium.

Production Example 2

Honggukgyun with Monascus pilosus Except for using IFO 4480 strain (which is the same strain as KCCM 60396 strain) in the same manner as in Preparation Example 1, except that Monascus pilosus A culture solution was obtained.

Production Example 3

Honggukgyun with Monascus ruber Except using KCCM 60617 strain is Monascus and for seed in the same conditions and method as in Preparation Example 1 ruber A culture solution was obtained.

Production Example 4

Aspergillus with similar characteristics to tereus ATCC 21542 strain was cultivated in potato dextrose agar medium (PDA medium) for 7 days and activated. Aspergillus tereus An agar block containing the bacteria was cut out. Thereafter, the cut agar block was added to 100 ml of an optimal gold culture medium (1.0 g of red bean powder, 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO ₄ , 100 ml) and incubated at a stirring speed of 150 rpm at about 28 ° C for 10 days to obtain Aspergillus tereus To obtain a culture medium. Then, the Aspergillus tereus culture was stirred with a blender to produce Aspergillus tereus Mycelial mycelium was homogenized and Aspergillus tereus A culture solution was obtained.

3. Production of Persimmon Fermented Tea

Production Example 5

After removing persimmon leaves from persimmon leaves, leaves with diseased leaves and insects were removed and persimmon leaves were selected. The selected persimmon leaves were washed three times with tap water, steamed for about 2 minutes in a steamer, and then boiled and dried. The dried persimmon leaves were cut to the appropriate size. Was placed into a flask of 500㎖ a persimmon leaf cutting capacitor 10g, optimal gold medium here (red bean powder 1.0g 4.0g glucose, peptone _{1.5g, KH 2 PO 4 0.4g,} MgSO 4 · 7H 2 O 0.1g, NaH 2 0.1 g of PO ₄ , and 100 ml of distilled water) was added to moisten the persimmon leaves moistly. Then, the flask was sealed, transferred to an autoclave, and sterilized by treating it at about 121 DEG C for 15 minutes. The sterilized persimmon leaves were inoculated into a flask containing Monascus purpureus 0.5 ml of the culture was inoculated and cultured at about 28 ° C for 10 days to ferment the persimmon leaves. Fermented persimmon leaves were dried at 50 ° C for 5 hours to prepare persimmon leaf fermented tea.

Production Example 6

As the seed bacterium, Monascus for seeds obtained in Production Example 2 pilosus The fermented tea of the persimmon leaves was prepared under the same conditions and in the same manner as in Preparation Example 5, except that the culture solution was used.

Production Example 7

As the seed bacterium, Monascus for seedlings obtained in Production Example 3 ruber The fermented tea of the persimmon leaves was prepared under the same conditions and in the same manner as in Preparation Example 5, except that the culture solution was used.

Production Example 8

Aspergillus for seeds obtained in Production Example 4 tereus The fermented tea of the persimmon leaves was prepared under the same conditions and in the same manner as in Preparation Example 5, except that the culture solution was used.

Production Example 9

As the seed bacterium, Monascus for seeds obtained in Production Example 2 pilosus The persimmon fermented tea was prepared under the same conditions and in the same manner as in Preparation Example 5 except that the culture was inoculated in an amount of 1 ml per 10 g of the dried persimmon leaf and that the fermented persimmon leaves were dried at 50 ° C for 10 hours.

Production Example 10.

As the seed bacterium, Monascus for seeds obtained in Production Example 2 pilosus Except that the culture broth was inoculated in an amount of 1 ml per 10 g of dried persimmon leaves, fermented at 25 ° C after seedling inoculation, and fermented persimmon leaves were dried at 50 ° C for 10 hours. Persimmon tea fermented tea was prepared.

Production Example 11.

As the seed bacterium, Monascus for seeds obtained in Production Example 2 pilosus Except that the culture broth was inoculated in an amount of 1 ml per 10 g of the dried persimmon leaf, the fermented product at 35 ° C after seedling inoculation, and the fermented persimmon leaf were dried at 50 ° C for 10 hours. Persimmon tea fermented tea was prepared.

Comparative Production Example 1

After removing persimmon leaves from persimmon leaves, leaves with diseased leaves and insects were removed and persimmon leaves were selected. The selected persimmon leaves were washed three times with tap water, steamed for about 2 minutes in a steamer, and then boiled and dried. The dried persimmon leaves were cut to the appropriate size to prepare persimmon leaves.

4. Evaluation of shape and function of persimmon leaf tea

(1) The form of fermented tea of persimmon leaves

FIG. 2 is a photograph of the persimmon fermented tea prepared in Production Example 6, and FIG. 3 is a photograph of the persimmon leaf tea produced in Comparative Production Example 1. 2, the persimmon fermented tea was obtained when the fermentation time was 6 days. As shown in FIG. 2 and FIG. 3, the persimmon leaf fermentation tea has Hong Guk-gyun on the surface of the persimmon leaf in the form of mycelium.

(2) Changes in the content of monacolin K in persimmon leaves

1 g of the persimmon fermented tea was pulverized, 20 ml of 70% ethanol aqueous solution was added thereto, and the mixture was extracted at about 25 ° C for 24 hours, followed by filtration through a filter paper to prepare an extract of persimmon leaf tea. The extract was analyzed by high performance liquid chromatography (HPLC) to determine the content of monacolin K. At this time, Mevinolin was used as a standard substance.

The content of monacolin K in the persimmon leaf tea fermented tea prepared in Preparation Example 6 and in the persimmon leaf tea prepared in Comparative Preparation Example 1 was analyzed to determine whether or not the fermentation produced monacolin K. 4 shows the HPLC analysis peak of the persimmon fermented tea obtained when the fermentation time was 6 days in Production Example 6, and Fig. 5 shows the HPLC analysis peak of the persimmon leaf tea prepared in Comparative Production Example 1. Fig. In Fig. 4, "MK" means Mona Colin K. As shown in FIG. 4 and FIG. 5, monacore K was present in the persimmon leaf tea fermented by Hong Guk Kyo, whereas monacore K was not detected in the non fermented persimmon leaf tea.

The content of monacolin K per g of the persimmon fermented tea prepared in Preparation Examples 5 to 8 was analyzed in order to examine the content of monacolin K in the persimmon fermented tea according to the type of strain.

Table 1 below shows the content of monacolin K in the persimmon fermented tea prepared in Preparation Examples 5 to 8. In the following Table 1, the persimmon fermented tea was obtained when the fermentation time was 6 days.

Persimmon leaves Monacoline K content (/ / g) The persimmon fermented tea of Preparation Example 5 295 The persimmon fermented tea of Preparation Example 6 289 The persimmon fermented tea of Preparation Example 7 206 The persimmon fermented tea of Preparation Example 8 291

Considering the content of monacolin K as shown in Table 1, the strain suitable for manufacturing the persimmon fermented tea was Monascus purpureus , Monascus < RTI ID = 0.0 > pilosus , and Aspergillus tereus .

In order to examine the content of monacolin K in the fermented tea of the persimmon leaves according to the fermentation temperature and fermentation time, the content of monacolin K per g of the persimmon fermented tea prepared in Preparation Examples 9 to 11 was analyzed. 6 is a graph showing the content of monacolin K in the persimmon fermented tea prepared in Production Examples 9 to 11 by fermentation time. As shown in FIG. 6, the optimum fermentation temperature was 28 ° C for fermentation time of fermented persimmon leaves of 7 days or less, and 25 to 28 ° C for fermentation time exceeding 7 days. It is judged that the fermentation temperature is 26 to 30 ° C.

(3) Changes in total polyphenol contents of persimmon leaves

To determine the content of total polyphenol compounds in the persimmon leaf fermentation tea according to the type of strain, the content of total polyphenol compounds per gram of persimmon leaf tea prepared in Preparation Examples 5 to 8 and Comparative Preparation Example 1 was analyzed Respectively. 1 g of the persimmon fermented tea was pulverized, 20 ml of 70% ethanol aqueous solution was added thereto, and the mixture was extracted at about 25 ° C for 24 hours, followed by filtration through a filter paper to prepare an extract of persimmon leaf tea. To 1 ml of the extract sample, 1 ml of Folin-Ciocalteau's phenol reagent (supplier: Sigma) and 1 ml of 10% Na ₂ CO ₃ solution were added in order, and the mixture was allowed to stand at room temperature for about 1 hour and then absorbance was measured at 700 nm. A standard calibration curve was obtained using gallic acid as a reference material, and the total polyphenol compound content was calculated in comparison with the standard calibration curve.

Table 2 shows the total content of polyphenol compounds in the persimmon leaf tea prepared in Preparation Examples 5 to 8 and the persimmon leaf tea prepared in Comparative Preparation Example 1. In Table 2, the persimmon fermented tea was obtained when fermentation time had passed three days.

Persimmon leaves Total polyphenol compound content (占 퐂 / g) The persimmon fermented tea of Preparation Example 5 59 The persimmon fermented tea of Preparation Example 6 64 The persimmon fermented tea of Preparation Example 7 57 The persimmon fermented tea of Preparation Example 8 63 The fermented tea of the persimmon leaves of Comparative Production Example 1 55

As shown in Table 2, when the total polyphenol compound content is considered, the strains suitable for manufacturing the persimmon fermented tea were Monascus pilosus and Aspergillus tereus .

(4) Changes in total flavonoid compound contents of persimmon leaves fermented tea

To determine the content of total flavonoid compounds in the persimmon leaf fermented tea according to the type of strain, the contents of total flavonoid compounds per g of the persimmon leaf tea prepared in Preparation Examples 5 to 8 and Comparative Preparation Example 1 were analyzed. 1 g of the persimmon fermented tea was pulverized, 20 ml of 70% ethanol aqueous solution was added thereto, and the mixture was extracted at about 25 ° C for 24 hours, followed by filtration through a filter paper to prepare an extract of persimmon leaf tea. 0.1 ml of a 10% aluminum nitrate solution, 0.1 ml of a 1 M potassium acetate solution and 4.3 ml of an 80% aqueous ethanol solution were added to 0.5 ml of the extract sample, and the mixture was allowed to stand at room temperature for about 40 minutes. Nm absorbance was measured. A standard calibration curve was obtained using quercetin as a standard substance, and the total flavonoid compound content was calculated in comparison with the standard calibration curve.

Table 3 shows the total flavonoid compound contents of the persimmon fermented tea prepared in Preparation Examples 5 to 8 and the persimmon leaf tea prepared in Comparative Preparation Example 1. In the following Table 3, the persimmon fermented tea was obtained when fermentation time had passed three days.

Persimmon leaves Total flavonoid compound content (占 퐂 / g) The persimmon fermented tea of Preparation Example 5 262 The persimmon fermented tea of Preparation Example 6 282 The persimmon fermented tea of Preparation Example 7 226 The persimmon fermented tea of Preparation Example 8 283 The fermented tea of the persimmon leaves of Comparative Production Example 1 260

As shown in Table 3, when the total flavonoid compound content was considered, the strains suitable for manufacturing the persimmon fermented tea were Monascus pilosus and Aspergillus tereus .

(5) Evaluation of Antioxidative Activity of Fermented Tea of Persimmon Leaf

The antioxidant activity of persimmon leaf tea was measured by radical scavenging activity by DPPH (1,1-diphenyl-2-picrylhydrazyl radical) assay. 1 g of the persimmon fermented tea was pulverized, 20 ml of 70% ethanol aqueous solution was added thereto, and the mixture was extracted at about 25 ° C for 24 hours, followed by filtration through a filter paper to prepare an extract of persimmon leaf tea. To 50 μl of the extract sample, 200 μl of 0.0002M DDPH solution (prepared by dissolving DPPH in ethanol) was added, mixed for 10 seconds, and reacted for about 20 minutes. Then, the absorbance of the reaction solution was measured at 517 nm using a spectrophotometer, and the DPPH radical scavenging activity was calculated by the following equation.

DDPH radical scavenging activity (%) = (1-A ₁ / A ₀ ) × 100

* A ₁ : Absorbance of the sample added with sample (ie liquid extract)

* A ₀ : Absorbance of sample without added sample

The DPPH radical scavenging activity of the persimmon leaf tea fermented tea prepared in Preparation Examples 5 to 8 and the persimmon leaf tea prepared in Comparative Preparation Example 1 are shown in Table 4 below. In Table 4, the persimmon fermented tea was obtained when fermentation time had passed three days.

Persimmon leaves DPPH radical scavenging activity (%) The persimmon fermented tea of Preparation Example 5 91.5 The persimmon fermented tea of Preparation Example 6 95.2 The persimmon fermented tea of Preparation Example 7 91.2 The persimmon fermented tea of Preparation Example 8 94.7 The fermented tea of the persimmon leaves of Comparative Production Example 1 88.4

As shown in Table 4, when the antioxidant activity was considered, the strains suitable for manufacturing the persimmon fermented tea were Monascus pilosus and Aspergillus tereus .

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Accordingly, the scope of protection of the present invention is not limited to the specific embodiments but should be construed as including all embodiments belonging to the claims attached hereto.

Claims (20)

Drying the persimmon leaves after boiling or growing them, and pretreating them;
The pre-treated persimmon leaves were seeded with Monascus husk pilosus) culture or Aspergillus terephthalate mouse (Aspergillus tereus ) culture broth and fermenting the same; And
And drying the fermented persimmon leaves.
[Claim 5] The method according to claim 1, wherein the pretreatment step further comprises adding a liquid medium or water to the dried persimmon leaf and sterilizing the persimmon leaf when the persimmon leaf is pretreated by steam-drying and drying.
[Claim 3] The process according to claim 2, wherein the amount of the liquid medium or water is 50 to 150 ml per 100 g of the dried persimmon leaf.
[Claim 3] The method according to claim 2, wherein the sterilization temperature and the sterilization time are 110 to 125 DEG C and 10 to 30 minutes.
3. The method according to claim 2, wherein the liquid medium comprises 3.0 g of red bean powder, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water (Kumhwa medium). ≪ RTI ID = 0.0 > 8. < / RTI >
The method according to claim 2, wherein the liquid medium is prepared by adding 1.0 g of red bean powder, 6.0 g of maltose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water (Kumhwa medium-modified). ≪ RTI ID = 0.0 > 21. < / RTI >
3. The method according to claim 2, wherein the liquid medium comprises 1.0 g of red bean powder, 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO _4, (Kumhwa medium-optimized). ≪ / RTI >
The method according to claim 1, wherein the monascus pilosus has an accession number of IFO 4480 or KCCM 60396.
[Claim 3] The method according to claim 1, wherein the Aspergillus tereus is ATCC 21542 .
The method according to claim 1, wherein said Monascus < RTI ID = 0.0 > pilosus) culture or Aspergillus terephthalate mouse (Aspergillus tereus culture medium was Monascus < RTI ID = 0.0 > pilosus) strain or Aspergillus terephthalate mouse (Aspergillus tereus) persimmon method for producing a fermented tea that a strain characterized in that obtained by inoculating and culturing in liquid medium.
11. The method according to claim 10, wherein the monascus < RTI ID = 0.0 > pilosus) culture or Aspergillus terephthalate mouse (Aspergillus tereus culture medium is characterized in that the mycelium of Monascus pilosus or Aspergillus tereus is homogenized.
11. The method according to claim 10, wherein the liquid medium is prepared by adding 3.0 g of red bean powder, 3.0 g of glucose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water , Kumgwa medium, 1.0 g of maltose, 6.0 g of maltose, 2.0 g of peptone, 0.8 g of KH ₂ PO ₄ , 0.05 g of MgSO ₄ .7H ₂ O, 0.1 g of NaCl and 85 to 110 ml of distilled water (Kumhwa medium-modified), 1.0 g of red bean powder, 4.0 g of glucose, 1.5 g of peptone, 0.4 g of KH ₂ PO ₄ , 0.1 g of MgSO ₄ .7H ₂ O, 0.1 g of NaH ₂ PO _4, And 85 to 110 ml of distilled water, or a medium in which two or more of them are mixed. The method of manufacturing a persimmon-fermented tea according to claim 1,
11. The method according to claim 10, wherein the liquid medium is a monascus saccharin pilosus) strain or Aspergillus terephthalate mouse (Aspergillus tereus ) is activated on a potato dextrose agar medium (PDA medium).
[Claim 5] The method according to claim 1, wherein the inoculation amount of the seed is 1 to 40 ml per 100 g of the dried persimmon leaves.
The process according to claim 1, wherein the fermentation temperature is 25 to 35 ° C and the fermentation time is 2 to 6 days.
A persimmon fermented tea produced by the method of any one of claims 1 to 15.
17. The fermented tea according to claim 16, wherein the content of the monacolin K in the persimmon fermented tea is 50 to 350 g per g of persimmon leaf fermentation tea.
17. The fermented tea according to claim 16, wherein the content of total polyphenol compounds in the persimmon leaf fermentation tea is 60 to 65 占 퐂 / g of persimmon leaf fermentation tea.
17. The fermented tea according to claim 16, wherein the total flavonoid compound content of the persimmon fermented tea is 275 ~ 285 g per g of the persimmon leaf fermentation tea.
[17] The persimmon leaf tea according to claim 16, wherein the radical scavenging activity of the persimmon leaf fermented tea by DPPH (1,1-diphenyl-2-picrylhydrazyl radical) assay is 92 to 97%.

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