KR101286068B1 - Method for selection of high-theanine containing tea tree - Google Patents

Abstract

The present invention relates to a method for selecting a high content of deanin from a population of tea trees, and according to the method of the present invention, it is possible to easily and quickly select an individual having a high content of deanine, thereby shortening a long breeding period. And can contribute greatly to the cultivation of varieties of tea trees.

Description

Method for selection of high-theanine containing tea tree}

The present invention relates to a method for selecting a high dean individual from a population of tea trees, and more specifically, (i) mixing an extract extracted from the tea tree species to a ninhydrin reaction solution; (ii) dropping the mixed solution on a thin layer chromatography plate or filter paper and confirming a color reaction; And (iii) selecting a tea tree and tree species that provide an extract having the darkest color according to the color reaction.

At present, there is a great deal of interest and support in the development of new varieties such as biodiversity conventions / genetic resources access and benefit sharing (CBD / ABS) and UPOV. Tea trees are no exception, and leading countries in the tea industry, including Japan, have long supported research on new breeding. The domestic car industry is expected to suffer severe tribulation due to tariff cuts on Chinese cars and increased imports following the FTA. In addition, the issue of royalty payments for tea trees imported from Japan is expected due to the entitlement of genetic resources of each country under the Convention on Diversity Convention (CBD).

Tea leaves contain caffeine, tannins, nitrogen, proteins, vitamins A and C, and inorganic salts, which are beneficial to the human body, such as awakening, diuresis, heart attack, detoxification, and fatigue recovery. Among them, deanine is the most amino acid contained in tea leaves. Deanine is one of the sedative and stress-reducing ingredients, and it promotes the production of alpha waves in brain waves. It also lowers blood pressure and helps other amino acids such as dopamine and GABA. Therefore, the growth of tea trees containing the above components is of great industrial value.

Korea has evolved into a unique group with various genetic variations, with the introduction of tea seeds 1200 years ago or sometimes live in the wild.However, tea trees grown in Korea are not fostered according to domestic cultivation characteristics and preferences, but are indiscriminate. The tea fields created by the real breeding by foreign varieties and unintroduced varieties of Korean varieties are not uniform, the quality is not good, and the native tea varieties indigenous to the cultivation environment of Korea are excellent. There is a problem that genes are being eroded. Therefore, it is urgent to select varieties with excellent genetic traits as indigenous to the climate climate of Korea and to register and disseminate farmland with unique varieties of Korea.

On the other hand, tea trees near Jirisan are well preserved in the wild due to regional isolation, and have a large number of genetic resources due to various genetic mutations. However, it is urgently needed to establish a selection method that can shorten the breeding period to within 7 to 10 years in the long-term breeding business plan, which takes 14 to 24 years in practice for breeding. In particular, in order to further increase the added value of tea, it is very important to establish a technology capable of selecting a tea tree with a high content of deanine, which is a high functional amino acid, from the tea tree population.

On the other hand, conventional plant-derived secondary metabolite content measurement has been made through the analysis of high performance liquid chromatography, GC, etc., these instrumental analysis method is complicated, time-consuming and expensive, and requires a lot of labor. While there has been a desire for a new selection method to replace this method, no effective method for screening high-denanine tea trees has been reported.

Accordingly, the present inventors have developed a colorimetric method for selecting high-denanine tea trees as a rapid and efficient selection method for the cultivation of varieties of tea trees, and applying them to extracts extracted from wild tea populations as a result of simple and rapid It confirmed that there is an effect of selecting, and came to complete this invention. Deanin high content selection method of the present invention can be used as a rapid and efficient selection method for the breeding of tea trees, it can shorten the traditional breeding period required 14-24 years will greatly contribute to the breeding of tea trees. Furthermore, the selection of high value-added deanine-containing tea trees will be able to cope with international movements such as CBD / ABS, and further contribute to raising incomes of farmers and fostering carmakers.

Accordingly, the present invention is to solve the problem that it was not easy to screen the tea tree with a high theanine content due to the complexity of the above-described deanine detection process, focusing on the chemical reaction of theanine in a low-cost and simple method The aim is to provide a method for screening high content of theanine.

In addition, an object of the present invention is to provide a tea product comprising an extract obtained from the tea tree having a high content of deanine selected by the above method as an active ingredient.

In order to achieve the above object. In one embodiment, the present invention comprises the steps of: (i) mixing the extract extracted from the tea tree species to the ninhydrin reaction solution; (ii) dropping the mixed solution on a thin layer chromatography plate or filter paper and confirming a color reaction; And (iii) selecting a tea tree and tree species that provide the extract having the darkest color according to the color reaction.

The tea tree extract is not limited thereto, but the tea tree extract may be obtained by crushing the tea tree species and distilled water into distilled water and extracting the hot water and filtering the extract with a filter paper. Preferably, the hot water extract may be extracted for 20 minutes to 40 minutes at 70 ° C. to 90 ° C., and 30 minutes at 80 ° C. in order to enhance the efficiency of color reaction.

In addition, the tea extract may have a concentration range of 100 to 1000 ppm. The color development can be easily confirmed with the naked eye in the concentration range, preferably when the 1000ppm is the most intense color reaction can be selected more easily.

The reaction solution for selecting the deanine content in the tea tree according to the present invention is characterized in that it comprises ninhydrin. In particular, the ninhydrin reaction solution may be used to dissolve 0.5-2mg of ninhydrin per 1mL of ethanol, it may be difficult to visually analyze because the color development is too light or too dark outside the mixing amount.

The mixing ratio between the tea tree extract obtained as described above and the reaction solution is preferably a ratio of the reaction solution: extract of 0.5-1: 1 (v / v), more preferably about 1: 1 (v / v), which is This is because the color is easily visible with the naked eye.

The step (ii) may further comprise a heat treatment for 10 minutes to 30 minutes at 60 ℃ to 80 ℃ before confirming the color reaction after the drop. The heat treatment may promote the reaction between the extract and ninhydrin.

The colorimetric method using ninhydrin used in the present invention uses the principle that when ninhydrin is reacted with an aqueous solution of amino acids, ninhydrin reacts with the hydroxyl group of the amino acid, and two molecules of ninhydrin are condensed by the action of amino acids. Therefore, since the color is unique according to the type of amino acid, it can be used for identification and quantification of amino acids and proteins.

The present inventors analyzed the 35 amino acid standards contained in the tea tree to establish the ninhydrin colorimetric method as a selection method of individuals containing a large amount of deanine from the population of the tea tree (see FIG. 1 (A) and Table 1). ), The optimum conditions were determined according to the ratio of deanine and ninhydrin (see FIG. 2). Based on this, the ninhydrin colorimetric method was applied to the tea extract to establish a selection method (FIG. 3). Reference). Then, as a result of performing ninhydrin color reaction with extracts of 160 wild tea trees in Hadong region, it was confirmed that the tea trees could be selected according to the contrast according to the different color development for each tea tree (see FIG. 4). . In addition, as a result of quantitative analysis of the deanine and amino acid content of the selected tea tree to test this result, it was possible to confirm the deanine content suitable for the results of the selection of high-density, low-containing individuals according to the chromaticity (Table 2 and Table 3). This was in agreement with the results of high performance liquid chromatography (HPLC) on the highest content of the dean and the lowest content of the dean. (See FIG. 1 (B) and FIG. 1 (C)).

As described above, according to the tea tree sorting method of the present invention, after visually judging the color reaction, the extract having the darkest color can be easily selected as a high deanine tea tree species.

The above-described method for selecting a high content of tea tree containing deanine can be equally applied to tea tree selection according to the content of other amino acids contained in tea tree.

Thus, as another aspect, the present invention comprises the steps of (i) mixing the extract extracted from the tea tree species to the ninhydrin reaction solution; (ii) dropping the mixed solution on a thin layer chromatography plate or filter paper and confirming a color reaction; And (iii) selecting the tea tree and the tree species that provide the extract having the darkest color according to the color reaction.

The amino acid is phosphoserine (Phosphoserine), phosphoethanolamine (Phosphoethanolamine), aspartic acid (Aspartic acid), serine (Serine), threonine (Threonine), asparagine (Glutamic acid), Glycine (Glycine) , Alanine, Valine, Cystine, Methionine, Isoleucine, Leucine, Tyrosine, Phenylalanine, Beta-alanine, β-Alanine, Gaba (GABA or γ-aminobutyric acid), histidine, carnosine, lysine, arginine, aminoisobutyric acid, citrulline, ornithine , And taurine may be selected from the group consisting of.

As another aspect of the present invention, there is provided a tea product comprising as an active ingredient an extract obtained from a tea tree having a high content of deanine selected by the above method. That is, by selecting the method of growing the tea tree species with a high deanin content in the tea tree by the above method, it is possible to obtain a tea product comprising a tea extract with a high deanine content obtained by processing it as an active ingredient.

As described above, according to the method of selecting the tea tree species with a high deanine content according to the present invention as described above, by applying a simple and rapid chemical color reaction method, by selecting an individual having a high deanine content, a highly functional amino acid Early selection of high indenine tea plants can be used for breeding.

1 (A) shows the chromatogram of 35 amino acid standards contained in the tea extract, (B) shows the chromatogram of the individual (H92) showing the most intense color reaction, (C ) Shows the chromatogram of the individual (H23) showing the lightest color reaction.
Figure 2 shows the degree of color development according to the concentration of deanine standard using the ninhydrin colorimetric reaction.
Figure 3 shows the degree of color development according to the concentration of tea extract using the ninhydrin colorimetric reaction.
FIG. 4 is a diagram illustrating various color reactions obtained from extracts of 160 Hadong tea trees using the ninhydrin colorimetric reaction, in which a box is shown a tea tree individual (dotted line) having a dark color reaction and a tea color individual (solid line). ) Is a selection.
Figure 5 is a graph showing the ratio of the deanine content to the total amino acid in each tea tree selected in Figure 4 above.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be obvious to you.

Example  1: Preparation of Tea Tree Extract

The tea leaves used in this experiment were taken 100 mg of leaves ( Camellia sinensis var. Sinensis) from Hadong area native wild tea from 1st to 3rd leaves of the shoots, dried, crushed and passed through 100 mesh. Vacuum packed in a bag and stored at -20 ° C. In the experiment, the powder was pulverized for 1 minute, mixed with 500 mg of leaves and 8 ml of distilled water, and extracted with hot water for 30 minutes in an 80 ° C water bath. The final extract was sufficiently cooled for 1 hour after the completion of extraction, and then filtered with distilled water to a final 10 ml (filter paper No., Advantec). Tea tree leaf extract of about 5% was obtained through the above method, and was used for the tea tree selection experiment having a high content of deanine.

Example  2: for selected objects Deni  And establishment of amino acid content analysis conditions

Amino acid analysis conditions were established for more accurate deanine content analysis in selected individuals for the verification of the ninhydrin colorimetric method, which allows early selection of high deanine individuals. Amino acid quantification was analyzed by HPLC by the method of Heresztyn et al. (2004).

Specifically, 10 g of 3% trichroloacetic acid (Sigma Co. USA) was added to 1 g of tea powder, followed by ultrasonic extraction at 30 ° C. for 2 hours (XD350H. Sungdong Ultrasound, Korea), followed by centrifugation at 4,500 rpm for 15 minutes. Was filtered through a 0.45 μm CA membrane filter (MFS-25, Advantec, MFS, Inc., CA, USA). 40 μl of filtered green tea extract solution was added to a small volume vial for HPLC (300 μl), and 50 μl of borate buffer and 10 μl of derivatization reagent (AccQ-Flour ™) were added for derivatization. Heat treatment was carried out for 10 minutes. 6-aminoquinoly- N -hydroxy-succinimidyl carbamate (AccQ-Flour) was used as a derivative reagent for fluorescence detection. The column used is analyzed by AccQTag ™ C ₁₈ Column (3.9 mm × 150 mm, 30 ° C., Waters Co. USA). The analysis conditions for mobile phases A and B were 0.1% sodium acetate (pH 6.8), 4% and 30%, respectively. A solvent containing% acetonitrile is used, and the gradient conditions are A: B = 94: 6 (20 minutes), 84:16 (20.2 minutes), 84:16 (35 minutes), 15:85 (38 minutes), 94: 6 (38.5 minutes) and 94: 6 (44.5 minutes), the flow rate was 1 mL / min, and the conditions of the fluorescence detector separated glutamic acid, γ-aminobutyric acid, arginine, and deanine at 250 nm and 395 nm. . The amino acid concentration as a result of the analysis was quantified using the Millenium 32 program. Standard amino acid (Sigma-Aldrich Chemical Co., Mo., USA) solution used for the analysis was made by the concentration curve to determine the amino acid content of each amino acid. Standards used are Phosphoserine, Taurine, Pho, Urea, Aspartic Acid, Hydroxyproline, Serine, Threonine, Asparagine, Glutamic Acid, Theanine, a-Aminoadipic, Glycine, Alanine, Citrulline, a-Amino-butyri, Valine, Cystine, Methionine, Isoleucine Leucine, Tyrosine, Phenylalanine, b-Alanine, B-Amino isobut, GABA, Histidine, 1-Methylhistid, 2-Methy Tryptophan, Carnosine, Ornithine, Lysine, Ammonia, Arginine.

The individual selected by the colorimetric method of the present invention was able to analyze the amino acid content, including deanine through the above analysis method. Chromatograms of a total of 35 amino acids are shown in FIG. 1. In addition, the analysis results of 35 amino acid standards are shown in Table 1 below, and as shown in Table 1, deanine was found at 38.37 minutes as the 11th material.

Analysis of 35 amino acid standards Name Ret . Time Area One Phosphoserine 3.760 2614848 2 Taurine 5.147 8094198 3 Pho 6.080 1150080 4 Urea 7.600 463023 5 Aspartic acid 18.320 3037322 6 Hydroxyproline 20.987 26477 7 Serine 24.747 2515610 8 Threonine 26.667 2569984 9 Asparagine 29.627 1518912 10 Glutathic acid 32.213 2558326 11 Theanine 38.373 20890466 12 a-Aminoadipic 40.587 2135449 13 Glycine 44.667 2589979 14 Alanine 47.013 2345570 15 Citrulline 49.093 2289461 16 a-Amino-butyri 51.573 2390338 17 Valine 55.893 2492907 18 Cystine 58.960 2789333 19 Methionine 62.400 2596521 20  Isoleucine 65.253 2533212 21 Leucine 66.907 2561485 22 Tyrosine 69.760 2608555 23 Phenylalanine 73.227 2443004 24 b-Alanine 75.013 458322 25 B-Amino isobut 75.707 480101 26 GABA 77.920 1798548 27 Histidine 82.667 2459132 28 1-Methylhistid 83.733 2244636 29 2-Methy 85.147 2028775 30 Tryptophan 86.987 1220409 31 Carnosine 88.347 1970072 32 Ornithine 90.693 2588350 33 Lysine 92.507 2456077 34 Ammonia 94.053 2391885 35 Arginine 101.653 2281518

Example  3: Ninhydrin  Colorimetric condition

In order to investigate the optimal colorimetric reaction between amino acids including deanine and ninhydrin, the conditions according to the ratio of deanine standard and ninhydrin were investigated. The ratio of deanine and ninhydrin was controlled by volume ratio 1: 1, 2: 1, 3: 1, 1: 2, 1: 3, respectively, and the aqueous solution of ninhydrin was 1mg / 1ml (1000 times). ) It was used by dissolving 20 ℃ of heat. Among these combinations, the purple reaction occurred best when the ratio of deanine and ninhydrin was 1: 1 in volume ratio. Therefore, in order to make the ratio of deanine and ninhydrin 1: 1, a treatment mixture of 10 μl of deanine + 10 μl of ninhydrin was added to a TLC plate, and then heat-treated at 70 ° C. in a dry oven for 10 to 20 minutes. At this time, since ninhydrin reacts with amino acids such as deanine, a UV reactant was used to generate a purple reactant having a maximum absorbance at 570 nm and 405 nm. This was confirmed as a useful way to check the height of the deanine content even with the naked eye.

As a result, as the concentration of deanine increased, the dark purple appeared, and color development was confirmed in the range of 100 to 1000 ppm concentration (see FIG. 2).

In other words, the method of the present invention can be measured by the naked eye and UV / VIS detector or 96 well UV reader through a simple reaction, it is possible to select a large number of samples in a short time at a low cost, it is possible to increase the breeding efficiency and shorten the time limit.

Example  4: in tea leaf extract Deni High content  Establish early individual selection

Tea leaf extract sample was prepared by the method of Example 1 to establish a deanin-containing individual early selection method using the tea leaf extract was used in the experiment by diluting the 5% extract prepared by the method 100-1000 times It was. In addition, the ninhydrin colorimetric method described in Example 3 was applied at a ratio of 1: 1 with the extract and ninhydrin.

As a result, as the concentration (100-> 1000ppm) was shown to show a deep purple reaction, it was confirmed that it is very suitable for the selection of high content of deanine using the tea leaf extract (see Fig. 3).

Example  5: from wild tea Colorimetric  Used amino acids and Deni High content  Object selection

In order to confirm whether the method mentioned in Example 3 is applicable to the actual tea tree, a test was performed using extracts extracted from 160 wild tea trees in Hadong area as samples. The 160 individual tea extracts prepared by the above method for deanin derivatization were prepared at 1000 ppm, which showed the deepest purple reaction, and reacted with the ninhydrin reagent (method described in Example 3).

As a result, the ninhydrin colorimetric reaction showed a different pattern for each of 160 extracts of tea trees, which could be easily selected according to the color contrast. In the present invention, according to the colorimetric reaction was classified into high amino acid and low amino acid containing deanine. The most prominent purple reaction among 160 individuals was estimated to be 143, 136, 126, 92, 81, and 15, and the most purple reaction among the 160 individuals was light An estimated low number was 156 and 23.

Example  6: Ninhydrin Colorimetrically  Of detailed tea tree objects Deni  Content test

The selected individuals were classified into high content (H15, H81, H92, H126, H136, H143 ) and low content (L23, L156) according to the degree of total amino acid content and their amino acid content including deanine was investigated. For the quantitative analysis of the deanine and amino acid content of the tea tree selected by the method described in Example 5, 10 ml of distilled water was added to 1 g of green leaves collected in a test tube, stirred at room temperature for 3 hours, and then centrifuged. (4 ° C 6,000 rpm, 15 min), only the supernatant was taken down into a flask and concentrated under a reduced pressure. The concentrate was dissolved in 5 ml of 0.2M Li-cirate buffer (pH 2.2), and analyzed after filtration of 0.2um membrance filter. Theanine and amino acid analysis was analyzed by the amino acid analyzer Syknm S7130 Amino Acid Reagent Organizer.

As a result, the deanin content of tea tree individuals selected through the ninhydrin colorimetric reaction was somewhat different, but the proper deanine content according to the selection of high and low content of deanine could be confirmed depending on the degree of chromaticity. (See Table 2 and Table 3).

The amino acid content of the six selected as high in theanine by the ninhydrin colorimetric method of the present invention reached 17.55 to 28.43 mg per 100 mg sample. However, the low content population had an amino acid content of 13.11 to 13.63 mg.

The proportion of deanine in the amino acid was higher in the high content population than in the low content population. H126 individuals had the highest content of deanine of 1.25 mg / 100mg among 28.43 mg / 100mg of amino acid. H92 is 1.53 mg / 100mg of the amino acid content of 25.93 mg / 100mg amino acid, H136 is 0.73 mg / 100mg of the amino acid content of 23.40 mg / 100mg amino acid, and H81 is 0.63 of 19.10 mg / 100mg amino acid content. The mg / 100mg, H15 was 0.25 mg / 100mg of the 18.63 mg / 100mg amino acid content, and the H143 was 0.15 mg / 100mg of the 17.55 mg / 100mg amino acid content.

In addition, L23 and L156 selected for the low content of deanine through the ninhydrin colorimetric method of the present invention showed a very low content of 0.03 mg / 100mg in the amino acid content of 13.11 mg / 100mg and 13.63 mg / 100mg, respectively. .

From these results, it is thought that the ninhydrin colorimetric method of the present invention can be applied to the selection of high and low amino acids, as well as selection according to deanine height.

On the other hand, even in the high and low content selection group, the amino acid type and content, including deanine, showed a difference between individuals. The difference in the content of H92 among the high deanine and L23 of the low deanine individuals was about 51 times, showing the largest difference in the deanine content (see FIG. 5).

In addition, as shown in (B) and (C) of Figure 1, even in the amino acid analysis conditions by HPLC shows the same difference as the result shown by the ninhydrin colorimetric method selection through the ninhydrin colorimetric method of the present invention Proved effective.

Deanine and Amino Acid Contents of Highly Selected Deanine Individuals amino acid High content  Population ( mg / 100 mg  sample) H15 H81 H92 H126 H136 H143 Phosphoserine 0.53 0.38 0.55 0.63 0.53 0.43 Phosphoethanolamine 0.03 0.03 - - - - Aspartic acid 3.18 3.10 3.03 4.48 3.88 3.50 Serine 0.40 0.43 1.48 0.58 0.53 0.48 Threonine 0.43 0.60 1.48 1.78 0.85 0.63 Asparagine 0.15 0.18 0.53 0.48 0.38 0.15 Glutathic acid 7.35 5.85 7.00 7.30 7.03 7.33 Theanine 0.25 0.63 1.53 1.25 0.73 0.15 Glycine 0.03 0.13 0.20 0.30 0.18 0.08 Alanine 2.78 2.58 3.18 4.03 3.50 2.10 Valine 0.25 0.28 0.93 0.88 0.58 0.13 Cystine 0.03 0.03 0.03 0.03 0.03 0.03 Methionine 0.03 - 0.08 0.08 0.05 - Isoleucine 0.13 0.10 0.30 0.28 0.23 0.05 Leucine 0.13 0.13 0.38 0.45 0.30 0.10 Phenylalanine 0.10 0.13 0.28 0.33 0.20 0.08 b-Alanine - - 0.03 0.03 - - GABA 2.50 2.00 3.68 4.13 3.23 1.58 Histidine 0.10 0.10 0.18 0.13 0.13 0.08 Carnosine 0.15 0.30 0.35 0.30 0.40 0.40 Lysine 0.08 0.08 0.25 0.20 0.15 0.05 Arginine - 0.08 0.08 0.15 0.15 0.10 Ammonia 0.05 1.03 0.18 0.13 0.13 0.08 Aminoisobutyric acid - 0.83 - - - - Citrulline - - - - 0.03 - Ornithine - - 0.03 0.05 0.03 - Taurine - - - - - - Tyrosine - 0.18 0.20 0.40 0.20 0.08 Urea - - 0.05 0.10 0.03 - Total amino acid content 18.63 19.10 25.93 28.43 23.40 17.55

Deanine and Amino Acid Contents of Selected Low Deanine Individuals amino acid High content population (mg / 100mg sample) L23 L156 Phosphoserine 0.45 0.55 Phosphoethanolamine - - Aspartic acid 2.68 2.85 Serine 0.35 0.48 Threonine 0.25 0.28 Asparagine 0.20 0.15 Glutathic acid 2.18 5.10 Theanine 0.03 0.03 Glycine 0.15 0.10 Alanine 3.30 1.18 Valine 0.48 0.28 Cystine 0.03 0.03 Methionine - - Isoleucine 0.08 0.13 Leucine 0.10 0.10 Phenylalanine 0.08 0.10 b-Alanine - - GABA 1.98 1.50 Histidine 0.08 0.08 Carnosine 0.33 0.33 Lysine 0.05 0.10 Arginine 0.08 0.08 Ammonia 0.15 0.08 Aminoisobutyric acid - - Citrulline - - Ornithine - - Taurine - - Tyrosine 0.08 0.10 Urea - - Total amino acid content 13.11 13.63

Claims (10)

(a) grinding tea tree species and extracting hot water at 70-90 ° C. for 20-40 minutes, and filtering the extract with filter paper to obtain an extract;
(b) reacting the extract obtained in step (a) by mixing the ninhydrin reaction solution with a volume ratio of 0.5 to 1: 1;
(c) dropping the reaction mixture of step (b) onto a thin layer chromatography plate or filter paper; And
(d) screening the darkest extract using a UV / VIS detector;
Deanine and amino acid-containing tea tree selection method comprising a. delete delete The method of claim 1, wherein the tea extract is characterized in that the concentration of 100 to 1000ppm, deanine and amino acid-rich tea tree selection method. The method according to claim 1, wherein the ninhydrin reaction solution is obtained by dissolving 0.5 to 2 mg of ninhydrin per 1 mL of ethanol. delete According to claim 1, wherein the step (c) further comprises the step of heat treatment for 10 to 30 minutes at 60 ~ 80 ℃ before confirming the color reaction after the drop, deanine and amino acid high tea Screening method. A tea product comprising as an active ingredient an extract obtained from a tea tree with a high content of deanine and amino acid selected by the method according to any one of claims 1, 4, 5 and 7. delete The method of claim 1, wherein the amino acid is phosphoserine, phosphoethanolamine, aspartic acid, serine, threonine, asparagine, glutamic acid. ), Glycine, alanine, valine, cystine, methionine, isoleucine, leucine, tyrosine, tyrosine, phenylalanine, betaalanine β-Alanine, GABA or γ-aminobutyric acid, Histidine, Carnosine, Lysine, Arginine, Aminoisobutyric acid, Citrulline Ornithine (Ornithine), and taurine (Taurine), characterized in that at least one selected from the group consisting of, deanin and amino acid-rich tea tree selection method.

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