JP2008263831A - Functional fermented tea and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide functional fermented tea containing new bioactive component through the fermentation treatment of tea leaves with microorganisms. <P>SOLUTION: A method for producing the functional fermented tea comprises inoculating to tea leaves, microorganisms which belong to Aspergillus or Eurotium, and produce 4-ethenyl-1,2-dimethoxybenzene, aceto-eugenol or iso-eugenol through fermenting tea leaves, and fermenting the product so as to produce fully fermented tea leaves which contain 4-ethenyl-1,2-dimethoxybenzene, aceto-eugenol or iso-eugenol. As a result of this, the functional fermented tea containing the useful components which are not contained in conventional tea is provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a post-fermented tea having a new function and a method for producing the same. More specifically, the present invention relates to 4-ethenyl-1,2-dimethoxybenzene, acetoate by fermenting tea leaves with a microorganism belonging to the genus Aspergillus or Eurotium. The present invention relates to a method for producing functional fermented tea that produces fermented tea leaves containing eugenol or isoeugenol, and functional fermented tea.

  The functional fermented tea of the present invention is characterized by fermenting tea leaves with microorganisms of the genus Aspergillus and Eurotium deposited at the Patent Microorganism Depositary Center, thereby providing antibacterial, neuroanesthetic, tonic stimulation, A novel functional fermented tea containing the above-mentioned physiologically active compound having an action such as sedation can be produced and provided. Conventionally, fermented tea has been used as a herbal medicine having various effects, for example, but it has not been easy to create tea leaves containing a component having a new bioactive effect by fermentation treatment. The present invention makes it possible to solve these problems of the prior art, and has functionality that contains physiologically active ingredients such as sedative effects that were not contained in conventional green tea and fermented tea. It provides new technology that enables the production of fermented tea.

  Traditionally, tea making consists of the steps of heating, twisting, and drying. From the difference in the manufacturing method, at which stage the tea leaves are naturally fermented by heating, and whether to ferment by mold. It is roughly classified into non-fermented tea (green tea), semi-fermented tea (Oolong tea, Baked tea), fully fermented tea (black tea) and post-fermented tea (black tea, puer tea). Non-fermented tea is obtained by steaming or heat-treating fresh tea leaves and killing them, then twisting them and drying them, and the enzymes that the fresh tea leaves themselves have lost. On the other hand, the semi-fermented tea and the completely fermented tea are those which are dried without sterilization, after having been fermented by the action of the enzyme group of the fresh tea leaves themselves, then inactivated by heating. Yes, it can be divided into semi-fermented tea and completely fermented tea depending on the degree of fermentation.

  Post-fermented tea is usually made from tea leaves that have been subjected to blue-killing treatment, planted with mold, deposited, and fermented using mold, and those that have been fermented for a long time are prized. The characteristics of post-fermented tea are said to be that catechins having a gallate group in tea leaves are decomposed to produce gallic acid having a strong antioxidant effect, and that umami and richness are increased. . Furthermore, post-fermented tea makes it possible to provide tea leaves with a unique fragrance or enhanced functionality through post-fermentation, and to provide functional foods using these functionalities. It is also possible to do. For example, Pu'er tea is a type of black tea, a post-fermented tea made by fermenting green tea with mold for a period of six months to one year or more, and has the effect of reducing cholesterol and neutral fat in the blood. It is said to be useful for effects and immune enhancement.

  As a known method for producing post-fermented tea, for example, the tea leaves are fermented with microorganisms of the genus Aspergillus or Rhizopus in the coexistence of tea stems, thereby maintaining the physiological activities such as cholesterol-lowering action of black tea. Thus, a production method has been proposed in which black tea can be produced in a short period of time and the yield of the extract from fermented tea is improved (see Patent Document 1). In addition, slurry fermentation of tea leaves with Aspergillus niger is carried out, the tea leaf suspension at the end of fermentation is collected and dried, and the amount of catechins having a gallate group per unit tea leaf can be extracted. A post-fermented tea beverage that has a low bitter taste and a strong umami and richness despite the fact that the amount of gallic acid and catechins extracted is large compared to other tea beverages (Patent Literature). 2).

  Further, as conventional techniques, 25 to 80 wt% water is contained in tea leaves after killing blue, tea leaves after rough softness, tea leaves after softness, tea leaves after neutralization, tea leaves after refinement, or rough tea. A fermented functional tea obtained by adjusting and fermenting by the action of yeasts or fungi such as shochu for shochu (A. sirousamii) and then drying by heating has been proposed. This fermented tea can be enjoyed with aroma, and at the same time, the functional substances are increased by leaving tea leaves-derived theanine and catechins in the tea leaves without decomposing them, and the glutamic acid contained in the tea leaves is changed. Containing γ-aminobutyric acid (see Patent Document 3).

  In addition, as an example of improving food quality and providing foods with new functions by using post-fermented tea as a food additive, Lactobacillus plantarum (lactic acid bacteria) isolated from green tea Fermented green tea that has been cultured, inoculated into fermented tea leaves and fermented, is easy to use in the food industry and easy to quality control, and can be used for drinking in the same way as ordinary tea leaves However, it is used to improve the flavor of bread as a lactic acid fermented species (see Patent Document 4), and is composed of a powdered or extracted microbial post-fermented tea obtained by fermenting green tea with black koji fungus, etc. A food additive having a flavoring action, a food preserving action and the like has been proposed (see Patent Document 5).

  As described above, post-fermented tea that has been developed in the past is a component that converts ingredients contained in tea leaves into useful substances by fermentation technology, and makes it possible to improve their aroma and functional characteristics. However, it has not been easy to develop and provide tea with new functions and scent components by conventional fermentation techniques.

JP 2005-333929 A JP 2005-278519 A JP 2006-14684 A Japanese Patent Laid-Open No. 11-276072 Japanese Patent Laid-Open No. 5-344852

  Under such circumstances, the present inventor has developed a method for producing post-fermented tea capable of efficiently producing post-fermented tea having new functional characteristics in a short period of time in view of the above-described conventional technology. As a result of earnest research, the production of post-fermented tea in the process of producing post-fermented tea produces a new fermented tea containing a new scent component and a new functional substance by carrying out the fermentation process using specific microorganisms. The present invention has been completed by finding that it is possible to do this and further research.

  An object of the present invention is to provide a post-fermented tea containing a novel functional substance as well as a novel aroma. Another object of the present invention is to produce post-fermented tea using a specific microorganism, thereby producing post-fermented tea containing 4-methyl-1,2dimethoxybenzene, acetoeugenol, or isoeugenol, which is a substance having a sedative effect. And a new post-fermented tea containing 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, 1-ethenyl-3-methoxybenzene. In addition, the object of the present invention is to provide a post-fermented tea in which the content of gallic acid, epicatechin and epigallocatechin is increased and the content of epicatechin gallate and epigallocatechin gallate is decreased compared to green tea. That is. Moreover, the objective of this invention is manufacturing and providing fermented tea after content of epigallocatechin and epicatechin increases compared with puer tea or 3 years black tea. Moreover, the objective of this invention is providing the manufacturing method of functional fermented tea which can manufacture and provide the post fermented tea containing a new useful component efficiently in a short period of time.

The present invention for solving the above-described problems comprises the following technical means.
(1) Belonging to the genus Aspergillus or Eurotium, inoculating tea leaves with microorganisms that ferment tea leaves to produce 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol. A method for producing functional fermented tea, comprising producing post-fermented tea leaves containing 1,2-dimethoxybenzene, acetoeugenol, or isoeugenol.
(2) The method for producing a functional fermented tea according to (1) above, wherein the microorganism belonging to the genus Aspergillus is PK-1 (FERM AP-21280).
(3) The method for producing a functional fermented tea according to (1), wherein the microorganism belonging to the genus Eurotium is Eurotium herbariorum.
(4) The method for producing a fermented functional tea according to (3) above, wherein the microorganism belonging to the genus Eurotium is KA-1 (FERM AP-21291).
(5) The above (1) to (4), wherein the tea leaves are selected from tea leaves after blue killing, tea leaves after rough softening, tea leaves after softening, tea leaves after middle softening, tea leaves after fine softening, or crude tea The manufacturing method of functional fermented tea of any one of these.
(6) The functionality according to any one of (1) to (5) above, which contains 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, or 1-ethenyl-3-methoxybenzene A method for producing fermented tea.
(7) A functional fermented tea produced by the method for producing a functional fermented tea according to any one of (1) to (6) above.
(8) A functional fermented tea comprising tea leaves containing 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol.
(9) The functional fermented tea according to (8) above, which contains 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, or 1-ethenyl-3-methoxybenzene.

Next, the present invention will be described in detail.
The present invention belongs to the genus Aspergillus or Eurotium, inoculates tea leaves with microorganisms that ferment tea leaves to produce 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol. A method for producing a functional fermented tea that produces post-fermented tea leaves containing -1,2-dimethoxybenzene (Benzene, 4-ethenyl-1,2-dimethoxy), acetoeugenol, or isoeugenol, And a functional fermented tea produced by this method.

  The present invention provides a novel functionality containing 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol that is not contained in tea leaves such as green tea and puer tea produced by conventional tea making methods. It is possible to produce and provide fermented tea. These compounds contained in the functional fermented tea of the present invention are components contained in, for example, essential oils (essential oils) and the like, and have antibacterial, antiviral, neural anesthesia, tonic stimulation, aphrodisiac, anthelmintic and the like. It is said that there is. In addition, these compounds are, for example, administered to a mammal (human) and sedated (see Patent Publication No. 2003-510365), mental sedative fragrance composition (see JP-A No. 2003-119490). Moreover, the usefulness as an active ingredient which exhibits a sedative effect in the throat sedative (refer patent publication 2001-526626) etc. is reported.

  In addition, the functional fermented tea of the present invention includes 2-ethyl-1-hexanol (1-Hexanol, 2-ethyl), 1-ethenyl-4-methoxybenzene (Benzene, 1-ethenyl-4-methoxy), or 1 -Ethenyl-3-methoxybenzene (Benzene, 1-ethenyl-3-methoxy) is contained. Since these aroma components are not contained in the conventional green tea, the present invention can provide a functional fermented tea that exhibits a fragrance that is not found in the conventional green tea.

  For the production of the functional fermented tea of the present invention, the microorganism PK-1 belonging to the genus Aspergillus (reception number FERM AP-21280), the europium hervariolam, or the microorganism KA-1 belonging to the genus Eurotium (reception number FERM AP- 21291) is characterized in that tea leaves are fermented, and functional fermented tea having the above components can be produced by the fermentation action of these microorganisms. These microorganisms are deposited at the Patent Microorganism Depositary Center (NPMD), microorganisms belonging to the genus Aspergillus are PK-1 (reception number FERM AP-21280), microorganisms belonging to the genus Eurotium are KA-1 (reception number FERM AP -21291). PK-1 belonging to the genus Aspergillus of the present invention is a microorganism collected from puer tea, and KA-1 belonging to the genus Eurotium is a microorganism collected from bonito.

  The tea leaves used in the production of the functional fermented tea of the present invention are not particularly limited, and any type of tea leaves, such as Ichibancha, Nibancha, Sanbancha, can be used as long as they are used in ordinary tea making. Even used. In order to produce the functional fermented tea of the present invention, it is preferable to heat-treat for the purpose of preventing an oxidation reaction or the like by an enzyme contained in the tea leaf before the fermentation treatment, and the harvested tea leaf Tea leaves that have been heat-treated may be used as they are, but the heating method is not particularly limited, and heating by various dryers using a direct fire method using a kettle, electricity, gas, etc., or steam is used. Any method or apparatus may be used as long as it can inactivate the enzyme by heat-treating tea leaves, such as a steaming and drying machine or a sun-drying machine. Examples of tea leaves whose enzyme has been deactivated by heating include, for example, tea leaves after killing in the conventional sencha production process, tea leaves after rough softening, tea leaves after softening, tea leaves after middle softening, tea leaves after fine softening, Or rough tea etc. are used for manufacture of the functional fermented tea of this invention.

  In order to ferment tea leaves, it is necessary to retain the raw tea leaves under conditions in which fermentation by the inoculated microorganisms proceeds. As the water content, tea leaves adjusted to contain about 15 to 80% by weight of water are used, and preferably 30 to 40% by weight are preferably used. Since it contains 25 to 40% by weight, the tea leaves after the middle meal are used for the fermentation treatment of the present invention without adjusting the water content. The fermentation temperature in the present invention is preferably about 15 to 40 ° C., and the fermentation treatment is usually performed continuously for 1 to 60 days, preferably 3 to 15 days. Therefore, it is shorter than the period required for efficient production. It is preferable to carry out by appropriately selecting the water content of the tea leaves, the fermentation temperature, the fermentation period and the like so that the production amount of the useful component of the present invention is increased as much as possible during the fermentation treatment.

  Next, after picking the most new shoots of tea leaves and carrying out steaming, rough koji, cocoon twisting, and mid-straw according to the standard tea making method, an appropriate amount of the microorganism PK-1 of the present invention is mixed with this green tea within a few days. The results of analyzing the contents of caffeine and catechins of fermented tea and green tea produced by culturing at about 25 ° C. for 1 to 4 weeks with stirring once are shown in FIGS.

  Analysis of the components contained in the produced fermented tea leaves revealed that there was no change in caffeine content between the fermented tea and green tea of the present invention, but the content of gallic acid was 10 times that of the fermented tea of the present invention compared to green tea. The above values were shown (see FIG. 2). Moreover, the content of epicatechin and epigallocatechin showed the tendency to increase by carrying out a fermentation process (refer FIG.5 and FIG.6). On the other hand, it was found that the contents of epigallocatechin gallate and epicatechin gallate are reduced in the functional fermented tea of the present invention as compared to green tea (see FIGS. 3 and 4). Thus, it turned out that post-fermented tea of this invention differs from green tea in content of a gallic acid and catechin. Gallic acid is a useful component that quickly exhibits an antioxidant / reducing action, prevents in vivo oxidation, and protects the living body from the damage of active oxygen. Therefore, the post-fermented tea of the present invention having an increased content of gallic acid is considered to exhibit excellent bioactivity.

  Comparing the content of catechins of commercially available post-fermented tea, puer tea or 3 year black tea, and green tea, epigallocatechin content in commercial post-fermented tea is about 1/10 that of green tea. It has been reported that the content of epicatechin is about 1/4 (see analysis values described in Table 3 of Patent Document 2). On the other hand, in post-fermented tea of this invention, as FIG.5 and FIG.6 shows, content of epigallocatechin and epicatechin is increasing rather than green tea. This indicates that the post-fermented tea produced by the fermenting action of the microorganism PK-1 of the present invention contains a fermentation product that could not be achieved by conventional puer tea or three-year black tea.

  Further, the aroma component of the functional fermented tea of the present invention produced by fermenting tea leaves using microorganism PK-1 (reception number FERM 21280) or microorganism KA-1 (reception number FERM AP-21291) is GC- As a result of analysis by MS, it was found that 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol was contained in the aroma component. These components are components that are considered to have antibacterial, antiviral, neural anesthesia, tonic stimulation, aphrodisiac, anthelmintic, sedative, and the like, and according to the present invention, contain components that exhibit an action effect not found in green tea. This makes it possible to produce and provide new fermented tea. Furthermore, the post-fermented tea of the present invention contains 2-ethylhexanol, 1-ethenyl-4-methoxybenzene, or 1-ethenyl-3-methoxybenzene, which is a component not contained in green tea. The analysis result of the aromatic component of the functional fermented tea and green tea of the present invention is shown in FIGS. The numbers attached to the peaks in each figure indicate that each peak corresponds to the compound described in Table 1 below.

  The extract obtained by extracting the functional fermented tea of the present invention with hot water produced by fermenting tea leaves with microorganism PK-1 or microorganism KA-1 exhibits a flower-like scent and a mild (less bitter) taste. It shows that the fermented tea of the present invention is excellent in palatability. Moreover, the fermented tea of the present invention contains useful components having various physiological activities, and the present invention makes it possible to produce and provide a new functional fermented tea excellent in palatability. is there.

Next, the microorganisms used in producing the functional fermented tea of the present invention will be described in detail. A method for separating microorganisms and the characteristics of the separated microorganisms will be described with respect to PK-1 belonging to the genus Aspergillus and KA-1 belonging to the genus Eurotium.
A. About microorganism PK-1 Separation of Microorganisms from Pu'er Tea A medium composed of 1 g of matcha tea, 2 g of agar and 100 mL of water was sterilized by autoclaving at 121 ° C. for 20 minutes and used. 1 g of Pu'er tea was put in 10 mL of sterile water, and 0.1 mL of the resulting liquid was applied to the above flat medium. Subsequently, it culture | cultivated at 30 degreeC for 72 hours, the resulting colony was isolate | separated into the matcha agar slope culture medium, and preserve | saved, and this strain was set to PK-1.

2. Properties of the Isolated Microorganism A strain obtained by culturing the separated PK-1 under the following conditions was used as a test cell.
・ Media Potato Dextrose Agar “DAIGO” (Nippon Pharmaceutical, Tokyo) (PDA)
2% Malt Agar (MA)
Bacto Oatmeal Agar (Becton Dickinson, MD, USA) (OA)
LCA (Miura Medium) (LCA)
・ Culture temperature 25 ℃
・ Cultivation period 1 week 2.1 Macroscopic observation results Macroscopic observations were made after 1 week of culture on each culture plate, and the colony diameter, color (colony surface and back), surface properties, and presence or absence of soluble pigment production were recorded. did. As a result, the characteristics shown in Table 2 were observed on each plate after one week of culture.

2.2 Microscopic observation results
(1) Vegetative mycelium Mycelia were formed on or in the agar surface, and formation of colorless to brownish brown, septal hyphae was observed.
(2) Asexual reproductive organs a) Conidial head and stalk The conidial head was dark brown to black, the shape was spherical at the beginning of culture, and a shape that split into a columnar shape with the passage of culture time was observed. The pattern (conidial pattern) was upright from the vegetative mycelium, non-branched, no partition, and the surface was observed to be smooth, colorless and slightly colored. The tip of the handle swelled and the formation of a crest was observed. The apical sac showed a spherical to subspherical shape. Formation of L-shaped to T-shaped pedicle cells was observed at the base of the handle.
b) Conidia-forming cells Cylindrical metre was formed in a row from the entire edge of the apical sac, and the structure of a two-row aspergillam was formed in which the conidia-shaped cell phialide was formed. The fiarride showed an ampoule shape with a short neck.
c) Conidia Conidia were fiaro-type conidia, spherical to subspherical, 1 cell, the surface was fine to spiny to rough, colorless to dark brown.
(3) Sexual reproductive organs The formation of sexual reproductive organs could not be confirmed from cultured specimens of about one month.

3. Estimation of belonging taxon of microorganism PK-1 obtained by the above isolation method The base sequence of 28S rDNA-D1 / D2 obtained from PK-1 is a reference strain NRRL4851 of Aspergillus phoenicis, an ascomycete fungus. It showed 100% homology with the base sequence of A. phoenicis including session number U28822) (see Table 3). In the phylogenetic tree prepared based on the obtained sequence, the same phylogenetic branches as A.phenicis, A.tubingensis, A.awamori and A.niger were formed (see FIG. 11). The ITS-5.8SrDNA base sequence obtained from PK-1 shows 100% homology with base sequences of Asperugillus phoenicis, A. tubingensis, A. awamori and A. niger, which are a type of ascomycete. It was. The same phylogenetic branch was formed also in the phylogenetic tree (refer to Drawing 12) created based on the obtained arrangement. From the results of the above two DNA base sequences, the specimen was determined to belong to Aspergillus.

However, in the base sequence analysis of 28S rDNA-D1 / D2, 100% agreement with A. phoenics (A. niger var. Phoenics) is indicated, and closeness with A. phoenics is estimated, but ITS-5. From the nucleotide sequence analysis of 8S rDNA, 100% coincidence with a plurality of species was seen, so the species level assignment could not be estimated. Therefore, PK-1 was estimated to be a kind of Aspergillus genus closely related to A. phoenics, A. tugingensis, A. awamori, and A. niger from the analysis results of both base sequences. All of these species considered to be closely related to PK-1 are included in the Nigri section, a member of the Asprugillus genus group, and are a group of bacteria commonly known as so-called black Asperigilli. .
On the other hand, as a result of observing colony properties and morphology, PK-1 formed black colonies, and the characteristics of the Aspergillus genus Nigri section were observed among Aspergillus genus such as apical sac and fiaro type conidia.

  From the results of the above 28rDNA-D1 / D2 and ITS-58SrDNA nucleotide sequence analysis, colony properties and morphology observation, PK-1 is A. phoenicis (A. niger var. Phoenicis), A. niger var. Presumed to be Aspergillus sp. closely related to tubingensis, A.awamori, and A.niger.

B. 1. Isolation and identification of bonito (microorganisms belonging to the genus Eurotium) from bonito Separation medium (MY20 medium)
Culture was performed using malt extract agar medium + 20% sucrose as a separation medium.
2. Separation of Bonito Mold 1 g of bonito slice was suspended in 10 mL of sterile water in MY20 flat medium, and 0.1 mL of the supernatant was applied to MY20 separation medium. This was cultured at 25 ° C. for 72 hours to isolate the resulting strain, planted in MY20 slant medium, and stored at 5 ° C.
3. Identification of strain KA-1 The resulting colony spreads rapidly on 20% scroll MY medium at 25 ° C., often irregular, the diameter after 14 days of culture is 5-7 cm, flat, pure green to gray-green conidial head Occurred in a wide band or spot. The back surface of the colony was brown, a large number of cones were formed, covered with loose yellowish-orange-red mycelium, spherical to subspherical, and the diameter was 75 to 100 μm. The cysts were spherical to subspherical, the diameter was 10 to 12 μm, the ascospores were lens-shaped, 4.8 to 5.6 × 4 to 5.5 μm, and lacked smooth surfaces, grooves and bumps. The conidia were grey-green, radial, the diameter was 500-1000 μm, and the conidia were iguriform oval-sub-spherical, 4.5-7 μm in diameter.
From the above observation results, the KA-1 strain was identified as Eurotium herbariorum.

The following effects are exhibited by the present invention.
(1) A post-fermented tea having a novel scent component and containing a novel functional substance can be provided.
(2) A novel post-fermented tea can be produced and provided by microorganisms that have not been used in conventional post-fermented tea.
(3) A post-fermented tea containing 4-methyl-1,2dimethoxybenzene, acetoeugenol, or isoeugenol, which is a substance having a sedative effect, can be produced and provided.
(4) A new post-fermented tea containing 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, 1-ethenyl-3-methoxybenzene as an aroma component can be produced and provided.
(5) Compared with green tea, fermented tea can be produced and provided after the content of gallic acid, epicatechin and epigallocatechin is increased and the content of epicatechin gallate and epigallocatechin gallate is decreased. .
(6) Compared with Pu'er tea and 3-year black tea, post-fermented tea having a composition with an increased content of epigallocatechin and epicatechin can be produced and provided.
(7) Post-fermented tea containing new components can be efficiently produced and provided in a short period of time.

  EXAMPLES Next, although this invention is demonstrated concretely based on an Example, this invention is not limited at all by the following Examples.

  In this example, a fermentation test of green tea using Aspergillus microorganism PK-1 (FERM AP-21280) was performed, and catechins and caffeine contained in the produced tea were analyzed and aroma was analyzed.

1. Materials and methods After picking the most new shoots from the tea garden (Yabukita) in Shimizu-ku, Shizuoka City, after steaming, rough kneading, twisting, and mid-brushing according to the standard tea making method, it is placed in a refrigerator (-20 ° C). Each 500 g was stored. 50 g of pre-cultured PK-1 was mixed with 500 g of green tea thawed at room temperature and cultured at about 25 ° C. for 2 weeks and 4 weeks with stirring once in several days (treatment 1). Thereafter, a part was freeze-dried and subjected to analysis of caffeine and catechins. The rest was stored frozen for scent analysis.
In addition, the precultured PK-1 is obtained by inoculating PK-1 in 50 g of medium green tea sterilized at 121 ° C. for 15 minutes in an autoclave and culturing at 30 ° C. for 7 days.

2. Analysis of caffeine and catechins About 25 mg of a finely pulverized sample was added 5 mL of acetonitrile, extracted by shaking for 1 hour, filtered, and subjected to analysis by HPLC.

3. Results and Discussion The caffeine content did not change even when fermented with microorganisms. However, gallic acid increased greatly by more than 10 times, and epicatechin gallate and epigallocatechin gallate gallates decreased. Like Chinese black tea made by fermenting black koji mold, it was estimated that gallate was decomposed during the fermentation process and gallic acid was produced in this experiment. Moreover, the amount of gallic acid produced was higher when the amount of PK-1 added was small and treated for a long time. Furthermore, when roasted, the content of gallic acid was greatly reduced. From this, it was found that increasing the amount of gallic acid is more effective when cultivating slowly and carefully, and it is more appropriate not to increase the drying temperature.

  On the other hand, epicatechin and epigallocatechin contents also tended to increase by fermentation, but not as much as gallic acid. Compared to normal values, the gallic acid content increased more than 5 times by fermentation.

In the same manner as in Example 1, the amount of PK-1 twice as much as that in Example 1 was added to green tea according to the standard tea making method and stirred for several weeks at about 25 ° C. for about one week. Each was cultured for 2 weeks (treatment 2).
Samples cultured for one week were partially lyophilized and subjected to analysis of caffeine and catechins. Some of the cultures for 2 weeks were dried in a dryer, and the rest were roasted on Japanese paper. A part of each was freeze-dried and subjected to analysis of caffeine and catechins. The rest was stored and subjected to aroma analysis.

  Inoculated into green tea that had been made up to the middle tea in accordance with the standard tea method in the same manner as in Example 1 except that the microorganism of the genus Europium (KA-1) separated from bonito was fermented at 25 to 35 ° C. By doing so, a post-fermented tea of the present invention was obtained. The content of caffeine and catechins in the produced post-fermented tea showed the same tendency as the result obtained in Example 1.

The aroma components of the fermented green tea prepared in Example 1 and Example 3 were analyzed.
How to collect aroma components:
About 150 mg of each tea leaf was put into a sample bottle using SPME (Micro Solid Phase Extraction Method) manufactured by Spelco, and about 3 mL of ultrapure water warmed in a range was added. One minute later, SPMA was set, and aroma components were collected by the headspace method using SPME while placing the sample bottle in hot water at 60 ° C. for 5 minutes, and subjected to GC-MS analysis. As the SPME fiber, a polydimethylkiloxane (= PDMS, methylsilicon) fiber having a membrane pressure of 100 μm was used.
Analysis conditions: column; TC-1 (nonpolar)
Vaporization chamber temperature: 250 ° C
Interface temperature: 230 ° C
Column temperature: 40 ° C .− (10 ° C./min)−200° C .− (40 ° C./min)−290° C. (2 min)
The analysis results are shown in FIGS.

  6 to 10 are compared, the functional fermented tea of the present invention shown in FIGS. 9 and 10 both have a peak of peak number 16, and the tea leaves produced by the conventional tea making method are used. Found that this peak number 16 did not exist. Therefore, the post-fermented tea of the present invention is 4-ethenyl-1,2-dimethoxybenzene (Benzene, 4-ethenyl-1,2-dimethoxy), acetoeugenol (Aceteugenol), or It was found to contain isoeugenol. The functional fermented teas of the present invention all have 10 peaks, and 2-ethyl-1-hexanol (1-ethyl), 1-ethenyl-4-methoxybenzene (Benzene) as aromatic components. , 1-ethenyl-4-methoxy), or 1-ethenyl-3-methoxybenzene (Benzene, 1-ethenyl-3-methoxy).

  As described above in detail, the present invention inoculates tea leaves with a microorganism that belongs to the genus Aspergillus or Eurotium and ferments tea leaves to produce 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol. A method for producing functional fermented tea, characterized by producing post-fermented tea leaves containing 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol. Functionality of fermenting tea leaves with PK-1 (FERM AP-21280) belonging to the genus Aspergillus or microbial KA-1 (FERM AP-21291) belonging to the genus Eurotium deposited at the Patent Microorganism Depositary It relates to a new technology for producing fermented tea.

  Conventionally, black tea, oolong tea, puer tea, black tea, and the like are well known as fermented teas, and the ingredients produced in the tea leaf fermentation process dissolve fat, regulate the intestine, suppress blood sugar, and enhance immune. It is said that it will demonstrate. The functional fermented tea of the present invention makes it possible to produce and provide a new tea leaf containing components not contained in conventional fermented tea or green tea, and has antibacterial action, neuroanesthetic action, sedative action The functional fermented tea containing the above-mentioned useful ingredients having physiological activities such as the above is provided. INDUSTRIAL APPLICABILITY The present invention provides a new type of fermented tea, thereby enhancing the utility of tea and is useful for providing a tea having an unprecedented taste and aroma.

The influence which the fermentation process by microorganisms has on the catechin content of green tea is shown. The influence which the fermentation process by microorganisms has on the gallic acid content of green tea is shown. The influence which the fermentation process by microorganisms has on the epicatechin gallate content of green tea is shown. The influence which the fermentation process by microorganisms has on the epigallocatechin gallate content of green tea is shown. The influence which the fermentation process by microorganisms has on the epicatechin content of green tea is shown. The influence which the fermentation process by microorganisms has on the epigallocatechin content of green tea is shown. The result of having analyzed the aromatic component of green tea by GC-MS is shown. The result of having analyzed the aromatic component of commercially available puer tea by GC-MS is shown. The result of having analyzed the aromatic component of the tea leaf fermented by PK-1 by GC-MS is shown. The result of having analyzed the aromatic component of the tea leaf fermented by KA-1 by GC-MS is shown. The molecular systematic diagram created by the neighborhood binding method based on the base sequence obtained from PK-1 is shown. A molecular system diagram created by the neighbor binding method based on the base sequence obtained from PK-1 is shown.

Explanation of symbols

About FIGS. 7-10 1-16: The peak number in each graph is shown, and the compound corresponding to each number is shown in Table 1.

Claims (9)

  It belongs to the genus Aspergillus or Eurotium, fermented by inoculating tea leaves with microorganisms that ferment tea leaves to produce 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol, and then ferment 4-ethenyl-1,2 -A method for producing functional fermented tea, characterized in that a post-fermented tea leaf containing dimethoxybenzene, acetoeugenol, or isoeugenol is produced.   The method for producing a functional fermented tea according to claim 1, wherein the microorganism belonging to the genus Aspergillus is PK-1 (FERM AP-21280).   The method for producing a functional fermented tea according to claim 1, wherein the microorganism belonging to the genus Eurotium is (Eurotium herbariorum).   The method for producing a fermented functional tea according to claim 3, wherein the microorganism belonging to the genus Eurotium is KA-1 (FERM AP-21291).   The tea leaves are selected from tea leaves after killing blue, tea leaves after softness, tea leaves after softness, tea leaves after middle softness, tea leaves after refinement, or crude tea. The manufacturing method of functional fermented tea of description.   The method for producing a functional fermented tea according to any one of claims 1 to 5, comprising 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, or 1-ethenyl-3-methoxybenzene.   A functional fermented tea produced by the method for producing a functional fermented tea according to any one of claims 1 to 6.   A functional fermented tea comprising tea leaves containing 4-ethenyl-1,2-dimethoxybenzene, acetoeugenol, or isoeugenol. The functional fermented tea according to claim 8, which contains 2-ethyl-1-hexanol, 1-ethenyl-4-methoxybenzene, or 1-ethenyl-3-methoxybenzene.

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