TW202131795A - Tea stalk containing gallic acid and methods of manufacturing the same - Google Patents

Abstract

Disclosed is a method of making tea stalk containing gallic acid. This method comprises the steps of planting tannase-producing bacteria into tea stalk, performing post-fermentation treatment on the tea stalk, and performing baking treatment on the tea stalk. It takes only half the time for manufacturing tea stalk containing a large amount of gallic acid by using the method of the present invention. Therefore, the economic benefits of the tea stalk can be much higher than that of tea leaf.

Description

Tea sticks containing gallic acid and preparation method thereof

This creation is related to a tea stick and its making method, and especially a tea stick containing gallic acid and its making method.

Gallic acid (chemical name: 3,4,5-trihydroxybenzoic acid) is an organic acid that can be found in plants such as gall, witch hazel, sumac, oak bark, tea, etc., also known as gallic acid or Gallic acid. Gallic acid has phenol and carboxylic acid, and is easily soluble in water, alcohol and ether. Gallic acid can synthesize gallic acid ester compounds, such as: methyl ester, ethyl ester, propyl ester, octyl ester, lauryl ester, stearyl alcohol ester, etc. These ester compounds are food antioxidants with excellent performance, especially propyl gallate as an antioxidant, which can be used for edible oils to prevent odor deterioration. Many research and scientific research results point out that gallic acid has various biological effects such as anti-inflammatory, anti-mutation, anti-oxidation, anti-free radical, anti-bacterial, anti-virus, etc. At the same time, gallic acid has anti-tumor effects and can inhibit mast cells. Tumor metastasis, thereby prolonging survival.

Park et al. found that after 24 hours of treatment with gallic acid and/or protease inhibitors, the growth of Calu-6 and A549 lung cancer cells was reduced. Yoon et al. found that gallic acid has pro-apoptotic and anti-inflammatory activities on fibroblast-like synovial cells (FLS) in rheumatoid arthritis, and can be used to treat rheumatoid arthritis in the future . Wang et al. found that gallic acid may have a protective effect on liver injury. The study by Chao et al. found that mice receiving oral gallic acid can resist hepatic steatosis, obesity and hypercholesterolemia caused by a high-fat diet. The study by Shahrzad et al. used tannin tablets (10% gallic acid and 90% glucose) and brewed black tea (containing 10% gallic acid, 93% of which is in the free state). ) To determine the pharmacokinetics of gallic acid and the relative availability of healthy humans. The results show that gallic acid is rapidly absorbed in the human body. Its half-lives are 1.19±007h for tablets and 1.06±006h for tea bags, which means The availability of gallic acid is similar between the two. The research results of Su et al. showed that gallic acid significantly inhibited melanin synthesis and tyrosinase activity in a dose- and time-dependent manner, and reduced the expression of melanin biosynthesis-related protein genes.

Oolong tea is a popular drink in China and Taiwan. It contains various biologically active ingredients, including polyphenols and phenolic acid. According to reports, old oolong tea has a better taste and better health benefits than fresh or fresh oolong tea. Generally, if the quality of tea is good enough, it is stored for more than five years and baked every year, it can be called old tea. The preparation of old oolong tea is a series of roasting fresh Tieguanyin oolong tea at 120-140°C for 72 hours, followed by long-term storage for 5, 10 and 20 years. Using liquid chromatography ultraviolet visible detection-tandemmass spectrometry to analyze the tea soup of old oolong tea, it was confirmed that the content of catechin (Catechin) in old oolong tea was reduced, while the content of gallic acid was relatively increased .

From the above research results, it can be seen that old tea containing gallic acid does have health benefits. However, the formation time of old tea is too long, and the content of gallic acid in various tea species is difficult to control; moreover, water-soluble tea polysaccharides The compound polysaccharides in the human body can significantly reduce blood sugar in the human body, and achieve the effect of preventing and treating diabetes without any side effects. Among them, the sulfated tea polysaccharide has a more obvious blood sugar lowering effect. It is expected to replace or cooperate with other diabetes drugs, and has a good market prospect .

In view of the above-mentioned problems with the knowledge and skills, one purpose of this creation is to provide a tea stick containing gallic acid and its production method.

To achieve the foregoing object, the present invention provides a method of making tea containing no gallic acid of the branch, comprising: a metal selected from aspergillus (Aspergillus), Mucor (Mucor), Actinomucor elegans (Actinomucor elegans) or China Rhizopus chinensis (Rhizopus chinensis) tannase-producing bacteria is inoculated into tea branches, wherein the aspergillus is selected from Aspergillus niger , Aspergillus japonicus , Aspergillus oryzae , Aspergillus sojae , or a combination thereof, and the Mucor is selected from Mucor hiemalis , Mucor silvaticus , Mucor prainii , or Mucor subtilissimus ; the tea branches inoculated with the tannase-producing bacteria are subjected to aerobic post-fermentation treatment at 25 to 50°C for 24 to 48 hours; and roasted at 70 to 150°C for post-fermentation The tea sticks are processed to obtain the tea sticks with gallic acid.

In one embodiment, the step of inoculating the tannase-producing bacteria to the tea branches includes: screening the tannase-producing bacteria with a plate medium; sequentially culturing the tannase-producing bacteria with a liquid medium and a fermenter, and then Take out the above-mentioned tannase-producing bacteria from the ferment tank; and evenly mix the above-mentioned tannase-producing bacteria and the above-mentioned tea sticks in sterile water.

In one embodiment, the weight ratio of the sterile water to the tea branch is 0.5 to 1:3.

In one embodiment, the weight of the gallic acid is preferably higher than 500 micrograms per milliliter of the extract obtained by extracting 2 grams of the tea sticks that have been roasted and post-fermented with 15 milliliters of water.

The present invention further provides a gallic acid-containing tea branch, which contains gallic acid and a tannase-producing bacteria, wherein the weight of gallic acid in each milliliter of extract obtained by extracting 2 grams of tea branch with 15 milliliters of water Preferably it is higher than 500 micrograms.

In one embodiment, the above-mentioned tannase producing strain selected from the genus aspergillus (Aspergillus) Mucor (Mucor), Actinomucor genus (Actinomucor) and Rhizopus (Rhizopus).

By inoculating the tannase-producing bacteria to the tea branches, and then performing post-fermentation and baking treatments on the tea branches, the tannase-producing bacteria can convert the tannic acid in the tea branches into gallic acid. The manufacturing method of the present invention It can quickly and stably make tea sticks with high content of gallic acid.

In order to enable Jun Shen to have a further understanding and understanding of the technical features of the present invention and the technical effects that can be achieved, the preferred embodiments and detailed descriptions are provided below.

In order to understand the technical features, content and advantages of this creation and its achievable effects, this creation is combined with the diagrams, and detailed descriptions are given in the form of embodiments as follows, and the diagrams used therein have only To illustrate and supplement the manual, it may not be the true proportions and precise configuration after the implementation of this creation. Therefore, the proportion and configuration relationship of the attached drawings should not be interpreted or limited to the scope of rights of the creation in actual implementation. In addition, in order to facilitate understanding, the same elements in the following embodiments are denoted by the same symbols.

Figure 1 shows the flow chart of the production method of tea sticks containing gallic acid in this creation. As shown in Fig. 1, the method for making tea sticks containing gallic acid includes: step 101, inoculating tannase-producing bacteria into the tea sticks; step 102, post-fermenting the tea sticks inoculated with tannase-producing bacteria Processing; and step 103, baking the tea sticks that have been fermented to obtain gallic acid-containing tea sticks.

In detail, step 101 includes the secondary steps of strain cultivation and tea branch inoculation. The sub-steps of strain cultivation in step 101 can be further divided into: strain screening, small-scale cultivation of strains, shake-flask cultivation of strains, and large-scale cultivation of strains.

Strain screening: Potato Dextrose Agar (PDA) plate medium was used to screen out the tannase-producing bacteria suitable for the growth of tea branches. The tannase-producing bacteria can be selected from the genus Aspergillus and Mucor. Mucor ), Actinomucor and Rhizopus . It is worth noting that the selected tannase-producing bacteria are all strains that meet food safety standards. Specifically, the genus Aspergillus includes but is not limited to: Aspergillus niger , Aspergillus japonicus , Aspergillus oryzae , Aspergillus sojae , or a combination thereof ; Mucor includes but is not limited to: Mucor hiemalis , Mucor silvaticus , Mucor praini , Mucor subtilissimus ; Actinomucor includes but is not limited to: Actinomucor elegans ; Rhizopus includes but is not limited to: Rhizopus chinensis , these strains can be obtained from American Type Culture Collection (ATCC) or Hsinchu Food Industry Research Institute.

Small-scale cultivation of strains: in an aseptic operation table, use aseptic technique to inoculate the selected tannase-producing bacteria to several plates of PDA plate medium, and then place the PDA plate medium in an incubator and cultivate for a period of time at an appropriate temperature The tannase-producing bacteria are grown to a basic number, for example, cultured at 20 to 40°C for 1 to 3 days, preferably, for example, 25 to 35°C for 1 to 3 days, and more preferably, for example, 30°C for 3 days.

Strain shake flask culture: in a sterile operating table, use aseptic technique to inoculate a small amount of cultured tannase-producing bacteria into a 500 mL triangular shake flask containing 100 mL of Potato Dextrose Broth (PDB) liquid medium Place the triangular shake flask in an incubator, and cultivate it at an appropriate temperature for a period of time to allow the tannase-producing bacteria to grow to a basic number, for example, at 20 to 40°C for 1 to 3 days, preferably, for example, at 25 to 35°C From 1 to 3 days, more preferably, for example, culture at 30°C for 3 days.

Large-scale cultivation of strains: use 4 shake flasks/5L fermenter as the required ratio of bacteria quantity, and centrifuge from the triangular shake flask with aseptic technique to take out the tannase-producing bacteria, and then inoculate the tannase-producing bacteria to the fermenter The tank is cultured in large quantities at an appropriate temperature, for example, culture at 20 to 40°C for 2 to 4 days, preferably, for example, at 25 to 35°C for 2 to 4 days, and more preferably, for example, at 33°C for 3 days.

The second step of tea branch inoculation in step 101 includes: centrifuging the tannase-producing bacteria of the aforementioned strains from the fermentation tank, and uniformly mixing the tannase-producing bacteria and the tea branches in sterile water to remove the tannase The producing bacteria are inoculated to the tea branches. During the inoculation, sterile water is added according to the weight of the tea branches. The weight ratio of the sterile water to the tea branches is, for example, 0.5 to 1:3, preferably, for example, 2 to 2.5 liters of sterile water is added per 6,000 g of tea branches. The amount of tannase-producing bacteria is adjusted according to the amount of tea branch inoculation, based on the weight that can be evenly mixed with the tea branch, the amount of inoculation per batch of tea branch is, for example, about 3,000 to 6,000 grams, and the type of tea branch is, for example, oolong tea Tea sticks.

Secondly, step 102 includes: performing aerobic post-fermentation on the tea branches inoculated with tannase-producing bacteria at an appropriate temperature, and the aerobic post-fermentation temperature is, for example, about 25 to 50°C, preferably, for example, about 33°C; The oxygen fermentation time is, for example, about 24 hours to 48 hours, preferably about 30 hours to 40 hours, and more preferably about 36 hours, for example.

Next, step 103 includes: baking the post-fermented tea branches at an appropriate temperature to remove the water content of the tea branches and increase the aroma components of the tea branches, so as to complete the production of the gallic acid-containing tea branches in this creation. In step 103, the baking temperature and time, taking Oolong tea sticks as an example, is, for example, about 70 to 150°C, preferably, for example, about 90 to 130°C, and more preferably, about 110°C, for example.

Please refer to Figure 2. Figure 2 shows the HPLC spectra of the tea sticks that have not been processed by the above-mentioned production process, the tea sticks that have been processed by the above-mentioned production process, and the gallic acid standard product.

First, take 2 grams of tea sticks that have not been processed by the production method of this creation (Comparative Example 1) and 2 grams of tea sticks that have been processed by the production method of this creation (Experimental Example 1), and extract them with 15 ml of hot water. And the treated tea sticks, and then use High Performance Liquid Chromatography (HPLC) to determine the gallic acid content of the untreated and treated tea stick extracts (5 or 10 microliters) quantity. This creation department uses Oolong tea as an example. Therefore, the above-mentioned tea branches are, for example, the oolong tea branches of Camellia sinensis , and the varieties of tea plants are, for example, but not limited to, Sijichun. That is, the method of making oolong tea sticks used in this creation is, for example, first fermented to make oolong tea, and then picked out the tea sticks from the tea pile for subsequent inoculation (planting), post-fermentation and drying. Therefore, the source of tea sticks is the same as that of tea leaves. The oolong tea sticks used in the data of this creation are, for example, but not limited to, purchased from Songfu Tea Factory, Mingjian Township, Nantou County, Taiwan. In addition, the tea sticks used and made in this creation are not limited to oolong tea sticks, and can also be tea sticks with different degrees of post-fermentation such as green tea or black tea.

HPLC specifications: pump L-2130, Autosampler L-2200, UV Detector L-2400. HPLC sample preparation 1. Weigh 2.0g tea branch sample into a 50mL centrifuge tube, add 15mL of hot water at 100°C, extract for 15 minutes, and centrifuge for 10 minutes (speed 4,000 rpm/min). 2. After centrifugation, take 1.2 mL of the supernatant in a 2.0 mL microcentrifuge tube and centrifuge for another 10 minutes (rotating speed 12,000 rpm/min). 3. After centrifugation, take 1mL of supernatant and pass through a 0.22m filter membrane (nylon). Chromatographic condition column is Inertsil ^® ODS-2 C-18 (4.6mm × 250mm), mobile phase (A) 0.026% phosphoric acid, (B) acetonitrile. Gradient conditions: 0-40 minutes, the concentration of B solution increased from 10% to 30%; 40-40.1 minutes, the concentration of B solution decreased from 30% to 10%; after 40.1 minutes, the concentration of B solution was maintained at 10% until the end of 70 minutes. Sample injection volume: standard 40L (gallic acid) (500g/mL), untreated tea branch sample 5 or 10L, and processed tea branch sample 5 or 10L; flow rate: 1mL/ min, absorbance value: 254 nm.

As shown in Figure 2, profile A is the HPLC profile of untreated tea branches, profile B is the HPLC profile of treated tea branches, and profile C is the HPLC profile of the gallic acid standard.

It can be observed from Atlas A and Atlas B that, compared with the tea sticks that have not been processed by the production method of this creation (Figure A), the GA content is greatly increased after the tea sticks are processed by the production method of this creation (Figure B). The specific components in Ning acid such as EGCG content and ECG content are slightly reduced. The measurement results show that the gallic acid content of the untreated tea branches is lower than the gallic acid content of the treated tea branches. From this result, it can be inferred that the specific components of EGCG and tannic acid in the tea branches are all of the present invention. The production method produces the source way of gallic acid.

This creation further analyzes the influence of different post-fermentation times on the gallic acid content produced by tea sticks. In this creation, the method of determining the content of gallic acid is to use HPLC analytical grade standard products, use the standard products to quantify the standard curve of gallic acid, and then use the interpolation method to obtain the content of gallic acid in each sample. Among them, this tea branch is the tea branch of Oolong tea.

First, take 2 grams of tea sticks produced according to the method of the present invention, extract the tea sticks processed by the production method of this creation with 15 milliliters of hot water, and then use high performance liquid chromatography (High Performance Liquid Chromatography, HPLC) to determine the Treat the amount of gallic acid contained in the extract of tea branches (5 or 10 microliters). This creation was repeated three times, and then the average value was taken. Figure 3 is a graph showing the concentration and time of gallic acid produced by tea sticks. It can be seen from Table 1 and Figure 3 that the content of gallic acid produced by tea branches will gradually increase with the post-fermentation time, and begin to rise rapidly at the 24th hour. When the post-fermentation time is about 36 hours, the gallic acid content will gradually increase. The acid content can reach the maximum. After that, the gallic acid content of the tea branches will gradually decrease with the post-fermentation time, and begin to decline rapidly at the 48th hour until it almost disappears. According to the production method of this creation, the content of gallic acid in the extract obtained by extracting 2 g of the tea branch of this creation with 15 ml of water is 322 to 527 μg/ml. For example, the weight of the gallic acid is preferably higher than 500 micrograms per milliliter of the extract obtained by extracting 2 grams of the tea branches that have been roasted and post-fermented with 15 milliliters of water.

Table 1 is the determination of the gallic acid content of tea sticks (unit: μg/ml) Second hour 0 twenty four 36 48 60 72 84 96 First measurement 32 330 517 423 122 37 57 16 Second measurement 28 326 549 441 115 35 19 16 The third measurement 36 310 516 240 79 43 14 3 average value 32 322 527 368 105 38 30 12 standard value 4 11 19 111 twenty three 4 twenty four 8

Figure 4 is a graph showing the concentration and time of gallic acid produced by tea. Among them, tea leaves are also processed by the method of making gallic acid-containing tea sticks in this creation. This tea is oolong tea, and the raw material source is the same as the tea sticks in Table 1. It can be seen from Table 2 and Figure 4 that the content of gallic acid produced by tea will also gradually increase with the post-fermentation time, but the increase is quite slow, and it starts to rise rapidly at the 36th hour, when the post-fermentation time is about 72 The gallic acid content can reach its maximum in hours. After that, the gallic acid content of tea sticks will gradually decrease with the post-fermentation time, and begin to decline rapidly at the 84th hour.

Table 2 shows the determination of gallic acid content in tea (unit: μg/ml) Second hour 0 twenty four 36 48 60 72 84 96 First measurement 52 180 434 910 1190 1030 1450 164 Second measurement 62 65 186 840 1067 1292 770 606 The third measurement 18 76 194 640 659 1079 939 612 average value 44 107 271 797 972 1134 1053 461 standard value twenty three 63 141 140 278 139 354 257

By comparing Table 1 and Table 2, it can be seen that according to the production method of this creation, the gallic acid content of tea sticks can reach the highest point after 36 hours of fermentation time, and the required fermentation time is only about 0.5 times that of tea. That is, the gallic acid content of tea requires 72 hours of post-fermentation time to reach the highest point. Although the gallic acid content of tea leaves can reach 2.15 times of the gallic acid content of oolong tea branches. However, in terms of raw material cost, taking Oolong tea as an example, the price of tea is 12 times that of tea sticks. In contrast, the economic benefit ratio of the tea sticks prepared by the production method of this creation compared with the tea leaves is 24:2.15.

In addition, as far as the storage method of post-fermentation products is concerned, tea branches only need to be stored in plastic bags at room temperature, while tea leaves need to be vacuum-packed in aluminum foil (antioxidants and desiccants are required). As far as product flavor and appearance are concerned, the appearance and flavor of tea sticks change less, and the change of tea leaves is large (the tea leaves are easy to unfold). As far as the stability of gallic acid is concerned, the reproducibility of tea branches is high, but the stability of tea leaves is low. In addition, the tea sticks can reach the highest gallic acid production within 36 hours, which can reduce the chance of contamination by bacteria. In contrast, it takes about 72 hours for oolong tea to reach the highest production of gallic acid, which will increase the chance of bacterial contamination. In addition, because the catechin content of oolong tea branches is low, it is less likely to inhibit the growth of tannase-producing bacteria. Therefore, the growth rate of tannase-producing bacteria is fast and it is easy to reach dominant strains. In contrast, oolong tea has a higher catechin content, which is easy to inhibit the growth of tannase-producing bacteria. Therefore, the growth rate of tannase-producing bacteria is slow and it is not easy to achieve a dominant strain.

Based on the above production method, this creation provides a gallic acid-containing tea branch, which contains gallic acid and tannase-producing bacteria, in which 15 ml of water is used to extract 2 grams of this creation’s tea branch. The content of gallic acid is 322 to 527 micrograms/ml. Tea sticks to the invention, tannase producing strain selected from the genus aspergillus food grade (Aspergillus), Mucor (Mucor), Actinomucor genus (Actinomucor) and Rhizopus (Rhizopus), and tea branch The raw materials can be selected from any tea tree (scientific name: Camellia sinensis ) oolong new tea, oolong old tea, green tea or black tea, and the tea branches are picked out after the tea making process is completed, and unnecessary tea branches are discarded.

This creation further uses HPLC-DAD to analyze the concentration of gallic acid components in freeze-dried tea leaves and tea branches before and after processing. For example, in this creation, the tea sticks can be extracted first, and then the drying step can be performed. The above-mentioned drying step is, for example, using traditional freeze-drying to obtain dry powder (without any excipients), wherein 1 ml of freeze-dried tea leaves and tea branch extracts can obtain approximately 0.04 g and 0.027 g of dry powder, respectively. Since the above-mentioned freeze-drying method can, for example, use a traditional technique, it will not be repeated here.

Table 3 is a table created by HPLC-DAD to analyze the concentration of gallic acid components in freeze-dried tea leaves and tea branches before and after processing. Among them, PFOTE is a processed tea leaf extract, OTE is an unprocessed tea leaf extract, PFVTE is a processed tea branch extract, OTE is an unprocessed tea branch extract, and GA is gallic acid. mg/g PFOTE OTE PFVTE VTE GA 28.35 + 3.48 1.10 + 0.58 19.53 + 0.69 1.19 + 0.15

In summary, this creation uses tannase-producing bacteria to inoculate the tea branches, and then the tea branches are subjected to 24-48 hours post-fermentation and baking treatment to make the tea branches produce gallic acid. The production method of this creation only takes half the time of tea to obtain tea branches with high content of gallic acid. Although the gallic acid content of tea branches is only half of the gallic acid content of tea, it is because of the price of tea branches. It is much lower than tea, and the time required to reach the highest gallic acid content is only half that of tea. Therefore, this creation can make the economic benefits of traditionally useless tea sticks far higher than tea.

The above descriptions are merely illustrative and not restrictive. Any equivalent modifications or alterations that do not depart from the spirit and scope of the present invention should be included in the scope of the appended patent application.

101, 102, 103: steps

Figure 1 is a flow chart of the creation method of making gallic acid-containing tea sticks.

Figure 2 shows the HPLC profile of the extracts of the treated tea tree ( Camellia sinensis ) oolong tea branch and the untreated oolong tea branch, and the gallic acid standard.

Figure 3 is a graph showing the concentration of gallic acid produced by the treated oolong tea branches and the fermentation time.

Figure 4 is a graph showing the concentration of gallic acid produced by treated oolong tea and the post-fermentation time.

101, 102, 103: steps

Claims (10)

The method of making tea containing no branches of gallic acid, comprising: genus selected from aspergillus (Aspergillus), Mucor (Mucor), Actinomucor elegans (Actinomucor elegans) or Chinese Rhizopus (Rhizopus chinensis) of a single Ning enzyme-producing bacteria are inoculated into tea branches, wherein the genus Aspergillus is selected from Aspergillus niger , Aspergillus japonicus , Aspergillus oryzae , Aspergillus sojae , or a combination thereof, and the Mucor is selected from Mucor hiemalis , Mucor silvaticus , Mucor prainii , or Mucor subtilissimus ; Perform aerobic post-fermentation treatment on the tea branch inoculated with the tannase-producing bacteria for 24 to 48 hours; and roast the tea branch after the aerobic post-fermentation treatment to obtain the gallic acid Tea sticks. According to the method for making gallic acid-containing tea branches as described in item 1 of the scope of patent application, the step of inoculating the tannase-producing bacteria to the tea branches includes: Screen the tannase-producing bacteria with a plate medium; After culturing the tannase-producing bacterium with a liquid medium and a fermenter in sequence, take out the tannase-producing bacterium from the fermenter; and The tannase-producing bacteria and the tea sticks are uniformly mixed in sterile water. The method for making gallic acid-containing tea sticks as described in item 1 of the scope of patent application, wherein the aerobic post-fermentation treatment time of the aerobic post-fermentation treatment step is 36 hours. According to the method for making gallic acid-containing tea sticks described in item 2 of the scope of patent application, the weight ratio of the sterile water to the tea sticks is 0.5 to 1:3. The method for making gallic acid-containing tea sticks as described in item 1 of the scope of patent application, wherein the tea sticks are oolong tea sticks. The method for making gallic acid-containing tea sticks as described in item 1 of the scope of patent application, in which 2 g of the gallic acid-containing tea sticks are extracted with 15 ml of water per ml of extract, gallic acid The content is 322 to 527 micrograms/ml. The method for making gallic acid-containing tea sticks as described in item 1 of the scope of patent application, wherein the processing temperature of the aerobic post-fermentation treatment step is 25 to 50°C, and the temperature of the baking step is 70 to 150°C . Tea containing sticks of gallic acid, comprising: gallic acid and is selected from the genus aspergillus (Aspergillus), Mucor (Mucor), Actinomucor elegans (Actinomucor elegans) or Chinese Rhizopus (Rhizopus chinensis) of Tannase-producing bacteria, wherein the genus Aspergillus is selected from Aspergillus niger , Aspergillus japonicus , Aspergillus oryzae , Aspergillus sojae , or a combination thereof, and the Mucor is selected from Mucor hiemalis , Mucor silvaticus , Mucor prainii , or Mucor subtilissimus . As described in item 8 of the scope of patent application, the gallic acid-containing tea branches, in which the content of gallic acid in each milliliter of extract obtained by extracting 2 g of the gallic acid-containing tea branches with 15 ml of water is 322 to 527 μg/ml. As described in item 8 of the scope of patent application, the tea branch containing gallic acid, wherein the tea branch is Camellia sinensis oolong tea branch.

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