WO2012046347A1

WO2012046347A1 - Tea extract

Info

Publication number: WO2012046347A1
Application number: PCT/JP2010/068214
Authority: WO
Inventors: 風雷陳; 川口　理衣; はるか木野; 冴美加東; 和種長野; 弘二村井; 怜藤田
Original assignee: 長谷川香料株式会社
Priority date: 2010-10-08
Filing date: 2010-10-08
Publication date: 2012-04-12
Also published as: CN103228147B; CN103228147A; JP5406379B2; HK1183207A1; JPWO2012046347A1; TWI404506B; TW201215330A

Abstract

The present invention provides a tea extract which comprises at least tannin, an amino acid and galacturonic acid, wherein galacturonic acid is contained in an amount of 1.1-5 mass% relative to the total solid content (in terms of Bx value) of the tea extract, the ratio of the content of galacturonic acid to the content of tannin (galacturonic acid/tannin) is 0.04-0.8 by mass, and the ratio of the content of galacturonic acid to the content of the amino acid (galacturonic acid/amino acid) is 0.08-0.8 by mass. The tea extract has masked bitter taste, is rich in sweet flavor, robust flavor and "umami" (tasty) flavor, and has a good flavor valance.

Description

Tea extract

The present invention relates to a tea extract having strong sweetness, richness and umami, and less astringency.

In recent years, products in which tea beverages are filled in cans or plastic bottles have been provided, and their production has been increasing steadily because of the high level of support from consumers' sweetness. As a recent trend, tea beverages with strong umami and richness and reduced astringency are preferred.
In the production of tea extracts, as a method of treating with an enzyme agent, for example, a method of extracting tea leaves using protopectinase and cellulase in combination (see Patent Document 1), a method of treating tea leaves with tannase (Patent Document 2) Cereals treated with pectinase, amylase and polyphenol oxidase (see Patent Document 3), impregnated with an aqueous solution of amylase, protease, cellulase or a mixed enzyme thereof, dried and then roasted at 100-170 ° C. Tea production method (see Patent Document 4), production method of instant tea extracted with a mixture of sticky starch and at least one enzyme selected from α- or β-amylase, cellulase and protease (see Patent Document 5) Digestion of tea leaves with tannase and at least one cell wall A method of moistening with an element (see Patent Document 6), a method of treating a tea leaf extract residue with cellulase and protease (see Patent Document 7), a method of pre-treating a hot water extract of tea with tannase and then freezing and concentrating it (Patent Document 6) Ref. 8), a method for producing a tea beverage with low turbidity by allowing chlorogenic acid esterase to act on tea extract (see Patent Document 9), and extracting tea raw materials in the presence of protease and tannase. A tea leaf extract comprising: a method for producing a tea extract (see Patent Document 10), an enzyme group containing at least cellulase, hemicellulase, pectinase and protopectinase; Production method (see Patent Document 11), tea leaves are extracted with water in the presence of protease, and the resulting extract is further purified with protease Extraction method of tea extract characterized by treatment (see Patent Document 12), decomposition of sugars such as glucoamylase, hemicellulase, pectinase, mannanase, invertase or α-galactosidase during and / or after extraction of tea raw materials A method for producing tea extracts characterized by enzymatic degradation using an enzyme (see Patent Document 13), and enzymatic degradation extraction treatment of tea raw materials using a bamboo shoot producing enzyme and cellulase, hemicellulase, pectinase or protopectinase A method for producing tea extracts (see Patent Document 14), which is characterized by the above, has been proposed.
However, these methods have achieved some results in terms of improving the taste such as sweetness, kokumi, and umami, and improving the yield, but there are still cell walls, proteins, etc. in the tea extraction residue. These useful ingredients remain, and not all of them are effectively used.

Japanese Patent Publication No.46-17958 Shoko 52-42877 Japanese Examined Patent Publication No. 62-15175 JP 57-47465 A Japanese Patent Publication No. 1-47979 Japanese Examined Patent Publication No. 4-63662 Japanese Patent No. 3157539 JP-A-5-328901 JP 11-308965 A JP 2003-144049 A JP 2003-210110 A JP 2008-67631 A JP 2008-86280 A JP 2008-125477 A

The object of the present invention is to extract cell wall components derived from tea leaves that could not be decomposed and extracted by the conventional enzyme-treated extraction method from tea leaves, and to further extract proteins that became extractable as the cell wall components were decomposed. By decomposing into amino acids, an amino acid component is extracted in abundantly, and as a result, a tea extract having abundant sweetness, kokumi and umami and less astringency is provided.

About 25% protein is contained in tea leaves (5 revised food ingredient table), and it is expected that a tea extract with a strong taste can be obtained by degrading this protein with protease. However, when only protease is allowed to act on tea leaves, not so many amino acids are released. In the previous study, the present applicant presumed that the protein in tea leaves was bound to tannin, and as a result of earnest research, the applicant extracted tea raw materials in the presence of protease and tannase. Has found that a tea extract with strong umami and richness and less astringency can be obtained, and has been proposed previously (see Patent Document 10).
However, even when the method described in Patent Document 10 is carried out, it has been clarified that cell wall components and proteins that have not been extracted still remain in the tea leaves after extraction. Therefore, the present inventors have studied to effectively utilize the cell wall components and proteins remaining without being extracted, and as a result, among pectinases, especially polygalacturonase efficiently decomposes the cell tissue of tea leaves. I found it. In order to decompose the pectic substances in tea leaves and increase the soluble solid content, it is usually considered that the active unit of polygalacturonase to be added should be increased. However, most commercially available pectinases do not have a very high activity unit of polygalacturonase, and are not effective at normal addition amounts (about 0.1 to 2% by mass with respect to tea leaves). The taste of the tea extract obtained is reduced by the excipients derived from the enzyme preparations and the enzyme protein, and unnatural sweetness that is different from that of tea is added. Problem arises.
Therefore, as a result of further studies to solve this problem, the present inventors have surprisingly found that tea leaves, in addition to protease and tannase, have a polygalacturonase activity of 20000 U / g or more. When an enzyme preparation having an amount of polygalacturonase activity added to an amount of 800 U or more with respect to 1 g of tea leaves is extracted, the yield of soluble solids from tea leaves is drastically improved, and a large amount of galacturonic acid is obtained. It was found that the amino acid yield was improved, and the resulting extract was found to have abundant sweetness, richness and umami, and the present invention was completed.
Thus, the present invention comprises at least tannin, amino acid and galacturonic acid,
(A) containing 1.1 to 5% by mass of galacturonic acid, based on the total solid content (Bx conversion) of the tea extract,
(B) the mass ratio of galacturonic acid / tannin is 0.04 to 0.8, and
(C) The present invention provides a tea extract characterized by having a mass ratio of galacturonic acid / amino acid of 0.08 to 0.8.

The tea extract of the present invention is obtained by converting about 40% by mass to about 80% by mass of the tea material used as a raw material into a soluble solid content, which greatly increases the extract yield from the tea material. It can be improved and contains a large amount of galacturonic acid. Moreover, the amino acid yield from tea raw materials can also be improved. Furthermore, the tea extract of the present invention contains abundant sweetness, kokumi and umami, and when added to tea beverages, it gives sweetness, kokumi and umami to tea beverages, or tea beverages. The sweetness such as kokumi and umami can be enhanced. In addition, when the tea extract of the present invention is produced by enzyme treatment of tea raw materials, the viscosity during enzyme treatment decreases with the enzyme treatment, and it becomes smoother, so the process of separating the tea leaf residue from the enzyme treatment slurry It can be done easily. Specifically, the time required for operations such as separation and filtration can be greatly shortened, the workability in production can be improved, and the production cost can be reduced by shortening the work time.

The tea extract of the present invention can be produced, for example, by subjecting tea materials to extraction treatment by adding protease, tannase, and an enzyme preparation having a polygalacturonase activity of 20000 U / g or more.
The above tea materials include fresh leaves obtained from buds, leaves, stems, etc. of tea (Camellia sinensis (L) O. Kuntze), which is an evergreen tree of the camellia family, non-fermented tea produced, and semi-fermented tea. Mention may be made of fermented tea. Examples of non-fermented tea include steamed non-fermented tea such as Sencha, Bancha, Hojicha, Gyokuro, Kabusecha, and Tencha, and unfermented tea such as Kama fried tea such as Ureshino tea, Aoyagi tea, and various Chinese teas. Examples of the semi-fermented tea include baked tea, iron kannon tea, oolong tea; and examples of the fermented tea include black tea, pu-erh tea, Awaban tea, and Goishi tea. In addition, tea obtained by adding unfermented tea or semi-fermented tea with flowers can be used. Among these, green tea, oolong tea, jasmine tea, and the like are preferable from the viewpoint of obtaining tea extracts having a fresh and natural aroma, sweetness, umami, and the like.
Proteases used for the above-mentioned enzyme treatment of tea raw materials are enzymes that hydrolyze peptide bonds of proteins and peptides. Such a protease is not particularly limited, and a protease derived from animals or plants or microorganisms can be used. For example, protease A “Amano”, protease M “Amano”, protease P “Amano” 3G, protease N “Amano” , Pancreatin F, Papain W-40, Promeline F (above, Amano Enzyme); Sumiteam (registered trademark) AP, LP, MP, FP, LPL (above, Shinnippon Chemical Co., Ltd.); Protin (registered) Trademark) FN (manufactured by Daiwa Kasei Co., Ltd.); Denapsin (registered trademark) 2P, Denateam (registered trademark) AP, XP-415, purified papain for foods, Biolase (registered trademark) XL-416F, SP-4FG, SP-15FG ( As described above, manufactured by Nagase ChemteX Corporation); Orientase (registered trademark) 22BF, 90N, ON 20A (above, manufactured by HIBI); Morsin (registered trademark) F, PD enzyme, IP enzyme, AO-protease (above, manufactured by Kikkoman Corp.); Sakanase (protease derived from Aspergillus manufactured by Kaken Pharma); Pandidase (registered trademark) NP-2, P, papain solver, protease YP-SS (manufactured by Yakult Pharmaceutical Co., Ltd.); Flavorzyme (registered trademark), Protamex (registered trademark), Neutase (registered trademark), Alcalase (registered) Trademark) (manufactured by Novozymes Japan); Cochlase (registered trademark) SS, P (and above, manufactured by Mitsubishi Chemical Foods); VERON PS, COROLASE PN-L, COROLASE N, COROLASE 7089, VERON W, VERON P (and above) Manufactured by AB Enzyme); Protin P, Deskin, Depi Chair, Protin A, Samoaase (registered trademark) (manufactured by Daiwa Kasei Co., Ltd.); Orientase (registered trademark) 90N, 10NL, 22BF, Nucleicin (registered trademark) (manufactured by HIBI); Aroase (registered trademark) AP -10 (manufactured by Yakult Pharmaceutical Co., Ltd.); Entilon NBS (manufactured by Toto Kasei Kogyo Co., Ltd.); Actinase (registered trademark) AS, AF (manufactured by Kaken Pharma); alkaline protease GL440, Purefect (registered trademark) 4000 L, protease 899, Protex 6L, Tasinase (registered trademark) (manufactured by Genencor Kyowa); other examples include animal-derived pepsin and trypsin. These proteases can be used alone or in combination of two or more. The amount of these proteases used varies depending on the titer, etc., and cannot be generally specified. However, it is usually about 0.01 U to about 100 U, preferably about 1 U to about 80 U per gram of tea raw material. it can.
Moreover, as tannase used for the enzyme treatment of said tea raw material, if it has the activity which decomposes | disassembles a tannin, there will be no restriction | limiting in particular and arbitrary things can be used. Specifically, for example, tannase-producing bacteria belonging to the genus Aspergillus, Penicillium, Rhizopus, Mucor and the like are obtained by solid culture or liquid culture according to a conventional method using a medium usually used for culturing these filamentous fungi. And a product obtained by purifying the treated product or its treated product by a conventional method. Commercially available tannase, for example, tannase “Kikkoman (5,000 U / g)” (Kikkoman), tannase “Kikkoman (500 U / g)” (Kikkoman), tannase (Mitsubishi Chemical Foods) Sumiteam TAN (manufactured by Shin Nippon Chemical Co., Ltd.) or the like may be used. These tannases can be used alone or in combination of two or more. The amount of tannase used varies depending on the titer, etc., and cannot be generally specified. However, the amount of tannase is usually about 0.1 U to about 50 U, preferably about 0.5 U to about 45 U per gram of tea raw material. it can.
In the present invention, in addition to the protease and tannase, an enzyme preparation having a polygalacturonase activity of 20000 U / g or more is preferably added in such an amount that the polygalacturonase activity is 800 U or more per 1 g of tea ingredients. Thus, the desired tea extract can be obtained. As a result, the yield of soluble solids from tea leaf materials is dramatically improved, and the resulting tea extract is rich in galacturonic acid and amino acids, and is rich in sweetness, kokumi and umami. A remarkable effect is obtained.
As described above, a technique for extracting tea raw materials by treating them with pectinase has been known before the filing of the present application. In addition, when pectinase is added to a tea material in addition to protease and tannase and extracted, a certain effect can be obtained as compared with the case where only protease and tannase are added and extracted. However, in addition to protease and tannase, in addition to protease and tannase, usually 800 U or more, preferably 1000 U or more, more preferably 1000 U to 10,000 U, and even more preferably 1500 U to 5000 U of polygalacturonase is added per gram of tea raw material. As a result, a surprising phenomenon occurs in which about 40% to about 80% by weight of the tea leaf material (dried tea leaves) is solubilized, and a large amount of galacturonic acid is produced with the decomposition of cell wall components. Furthermore, the amount of extracted amino acids also increased, and with these increases, it was found that umami, sweetness, kokumi, etc. were enhanced and a flavorful tea extract could be obtained in high yield.
Polygalacturonase is a kind of pectinase. Enzymes generally classified as pectinases include polygalacturonase, pectin lyase and pectin methylesterase. Polygalacturonase is an enzyme that hydrolyzes α-1,4 bonds in the main chain of polygalacturonic acid in pectin. Pectin lyase removes α-1,4 bonds in the main chain of polygalacturonic acid in pectin. Pectin methylesterase is an enzyme that hydrolyzes the methyl ester of pectin. Pectinase is an enzyme that is positioned at the center of an enzyme group that disrupts plant tissues. As described above, a technique for extracting tea raw materials by treatment with pectinase has been known before the filing of the present application. However, even when conventional tea pectinase described in, for example, the above-mentioned patent documents is used in a normal addition amount, tea material is treated with an enzyme, the tea tissue is sufficiently decomposed. That's not true. Therefore, we examined whether polygalacturonase, pectin lyase, or pectin methylesterase in pectinase is particularly effective against tea cell tissues. Polygalacturonase alone is also effective. Moreover, it discovered that sufficient decomposition | disassembly of a cell tissue was performed by using what has an active unit higher than conventionally used.
In the present specification, polygalacturonase activity is determined by allowing polygalacturonase to act on a polygalacturonic acid aqueous solution as a substrate by the Somogy Nelson method (J. Biol. Chem. 153, 375-380, 1994). The enzyme reaction product is a value measured by a colorimetric method for quantifying reducing sugar, and 1 unit of enzyme (1 U) means the amount of enzyme that produces 1 μmol of galacturonic acid per minute.
Examples of the pectinase include commercially available products such as pectinase PL “Amano”, pectinase G “Amano” (manufactured by Amano Enzyme), Pectinase-GODO (manufactured by Godo Shusei Co., Ltd.), sucrase (registered trademark) A, N , S (above, manufactured by Mitsubishi Chemical Foods), Sumiteam (registered trademark) AP-2, SPC, SPG, MC, PX, liquid Sumiteam AP-2 (above, manufactured by Shin Nippon Chemical Industry Co., Ltd.), pectinase XP-534 (Manufactured by Nagase ChemteX Corporation), Pectinex (registered trademark), Pectinex Ultra SP-L, Ultrazyme (registered trademark), Vinozyme (registered trademark), Citrozyme (registered trademark), Peelzyme (registered trademark) (above, Novonor Disk bioindustry); Cellulosin (registered trademark) PC5, PE60, PEL Soluble pectinase T (manufactured by HBI Enzymes Inc.), pectinase SS, pectinase HL (manufactured by Yakult Pharmaceutical Industry Co., Ltd.), and the like. Among these, as pectinase having particularly high polygalacturonase activity, for example, Sumiteam AP-2, SPC, SPG (manufactured by Shin Nippon Chemical Industry Co., Ltd.) can be mentioned.
The polygalacturonase activity of a general commercial pectinase preparation is usually about 500 U / g to about 20000 U / g. Therefore, in order to add 800 U to 1 g of tea leaf material, a large amount of pectinase preparation of 0.04 g to 1.6 g must be added to 1 g of tea leaf material. At that time, for example, if the amount of the enzyme preparation is added to 0.06 g or more, particularly 0.08 g or more with respect to 1 g of the tea leaf raw material, the influence of excipients and other components is strongly exerted on the tea extract, and the resulting tea There is a problem of adversely affecting the taste, for example, the taste of the fruit extract becomes light, an unnatural sweetness that is different from that of tea, or a miscellaneous taste is produced. Therefore, a pectinase originally having a high activity of 20000 U / g or more as the polygalacturonase activity can be used as it is, but in the case of a pectinase preparation having a polygalacturonase activity of less than 20000 U / g, for example, the enzyme It is necessary to purify the preparation by water miscible organic solvent (acetone, ethanol, etc.) precipitation, isoelectric point precipitation, ultrafiltration, gel filtration, etc., and collect and use fractions with polygalacturonase activity of 20000 U / g or more. There is.
In addition, in addition to the protease, tannase and polygalacturonase, in addition to the above-mentioned protease, a cellulase derived from Trichoderma longibrachiatum or Trichoderma reesei is added and extracted. Furthermore, the tea leaf tissue can be decomposed more efficiently and the extraction efficiency of the water-soluble component can be increased.
As described above, a technique for extracting tea by treatment with cellulase has been known before the filing of the present application. In addition, in addition to protease and tannase in addition to protease and tannase, extracted by adding cellulase derived from Aspergillus niger, Trichoderma viride, etc., add only protease and tannase Compared with the case, a certain effect can be obtained. However, when cellulase derived from Trichoderma longibrachiatum or Trichoderma reesei was extracted from tea materials in addition to protease and tannase, sufficient degradation of the cell tissue was performed. Turned out to be.
Examples of the cellulase derived from the above-mentioned Trichoderma longibrachiatum or Trichoderma reesei include, for example, cellulosin (registered trademark) T3 (manufactured by HIBI), Sumiteam (registered trademark) CS, C (or more). New Nippon Chemical Industry Co., Ltd.), Cellulase SS (manufactured by Nagase ChemteX Corporation), Sucrase (registered trademark) C (manufactured by Mitsubishi Chemical Foods), and the like. The amount of cellulase derived from Trichoderma longibrachiatum or Trichoderma reesei varies depending on the titer, etc., and cannot be generally stated, but is usually about 0.1 to about 0.1 g per tea raw material. Examples thereof include 200 U, preferably about 0.5 to about 100 U, more preferably about 1 to about 50 U.
In the present invention, other saccharide-degrading enzymes such as hemicellulase, protopectinase, glucoamylase, glucanase, mannanase, and α-galactosidase can be used in combination as long as the effects of the present invention are not hindered.
An embodiment for producing the tea extract of the present invention is exemplified as follows:
Prepare a solution in which 4 to 40 parts by weight of water and 0.1% to 1% by weight of ascorbic acid or sodium ascorbate of the tea raw material are dissolved as needed per 1 part by weight of the tea raw material, Tea raw materials are added thereto, and if necessary, sterilized at about 60 ° C. to about 121 ° C. for about 2 seconds to about 20 minutes, and then cooled. Next, tannase is first added and mixed uniformly, and then a protease and an enzyme preparation having a polygalacturonase activity of 20000 U / g or more are added to 800 U or more as polygalacturonase activity per 1 g of tea raw material. The enzyme treatment is performed at about 20 ° C. to about 60 ° C. for about 30 minutes to about 24 hours. After the enzyme treatment, the enzyme is inactivated at about 60 ° C. to about 121 ° C. for about 2 seconds to about 20 minutes, cooled, and separated using a suitable separation means such as centrifugation or filter paper filtration to obtain a clear tea extract. Obtainable. The obtained tea extract can be in the form of a concentrated solution by using an appropriate concentration means if desired.
The above enzyme-treated extraction produces about 4 to 5 times as much amino acid as tea extract without any enzyme treatment, and the cell tissue of tea materials decomposes to produce a large amount of galacturonic acid. About 40% by mass to about 80% by mass of tea used as a raw material can be converted into soluble solids.
As a result of increasing the solid content yield, the amino acid yield, and the galacturonic acid yield from the tea raw material by the above method, (a) galacturon based on the total solid content (Bx conversion) of the tea extract 1.1 to 5% by mass of an acid, (b) the mass ratio of galacturonic acid / tannin is 0.04 to 0.8, and (c) the mass ratio of galacturonic acid / amino acid is 0.08 to 0. .8 tea extract; preferably (a) 1.2 to 4% by mass of galacturonic acid based on the total solid content (Bx conversion) of tea extract, and (b) galacturonic acid / tannin And (c) a galacturonic acid / amino acid mass ratio of 0.14 to 0.6, more preferably (a) of a tea extract. Based on the total solid content (Bx conversion), galacturonic acid is 1.3 ~ Tea containing 3% by mass, (b) the mass ratio of galacturonic acid / tannin is 0.07 to 0.2, and (c) the mass ratio of galacturonic acid / amino acid is 0.19 to 0.4 Extract can be obtained.
In addition, galacturonic acid is thought to have effects such as masking of bitterness, masking of off-flavor, and imparting body feeling because it has a refreshing and sour acidity that makes you imagine high-quality tea such as matcha tea. The increase in galacturonic acid is presumed to be one of the important factors for the sweetness, richness and umami of the tea extracts of the present invention.
Thus, the present invention can provide, as one aspect, a tea extract in which the galacturonic acid in the tea extract is produced by enzymatic decomposition of the tea raw material.
If desired, the tea extract of the present invention can be stored for a long period of time by sterilization by heating after filling the container or before filling.
In addition, the tea extract of the present invention can usually be used in a liquid state as it is, but if desired, an excipient such as dextrin, modified starch, cyclodextrin, gum arabic or the like is added to the extract to form a powder. You can also
Hereinafter, the present invention will be described more specifically with reference to examples and comparative examples.

Reference Example 1 Measurement of polygalacturonase activity (Somogyelson method: see J. Biol. Chem. 153, 375-380, 1994)
0.1 ml of an appropriate dilution of the enzyme solution is added to 0.9 ml of 50 mM acetate buffer (pH 4.5) containing 1% polygalacturonic acid. After reacting the mixed solution at 45 ° C. for an appropriate (appropriate) time, the enzyme is inactivated by heating in a boiling water bath for 10 minutes, and ice-cooled to obtain a reaction solution. Add 0.3 ml of the somogenic copper reagent to 0.3 ml of the reaction solution, heat in a boiling water bath for 10 minutes, cool with ice, add 0.3 ml of Nelson reagent and stir well in a test tube mixer, and add 3 ml of ion-exchanged water. In addition, mix well with a test tube mixer. This solution is treated at 9000 rpm for 3 minutes in a centrifuge, and the absorbance (Abs.) At 500 nm of the supernatant is measured. On the other hand, using a solution obtained by inactivating an appropriate dilution of the enzyme solution in advance, the same operation as described above is performed to obtain the absorbance of the blank. From the enzyme concentration used, enzyme reaction time, and absorbance, the μmol number of galacturonic acid produced per minute per 1 g of enzyme is calculated, and the unit (U) per 1 g of enzyme is calculated.
Measured enzyme and polygalacturonase activity measurements:
Sumi Team AP2 (manufactured by Shin Nippon Chemical Industry Co., Ltd.): 12400U / g
Cellulosin PE60 (manufactured by HIBI): 20600 U / g
Sumiteam MC (manufactured by Shin Nippon Chemical Industry Co., Ltd.): 1690 U / g
Sucrase N (Mitsubishi Chemical Foods): 4550 U / g
Reference example 2
100 g of Sumiteam AP2 (manufactured by Shin Nippon Chemical Industry Co., Ltd.) (polygalacturonase activity by the above measurement: 12400 U / g) was dissolved in 1000 g of ion-exchanged water, and Vivaflow (registered trademark) 50VF05P2 (molecular weight cut off 30,000: Sartorius) And 30 ml of the non-passed part was recovered and lyophilized to obtain Reference Product 2 (12.0 g: polygalacturonase activity measured above: 86500 U / g).
Example 1
To a solution of 0.6 g of sodium ascorbate dissolved in 900 g of soft water, 100 g of green tea leaves (Chinese steamed blue) was sterilized at 80 ° C. for 5 minutes and cooled to 45 ° C. 1 g of tannase (Mitsubishi Chemical Foods Co., Ltd .: 500 U / g) was added thereto and stirred for 15 minutes. Thereafter, 1 g of protease M (manufactured by Amano Enzyme: 5500 U / g) and 4.8 g of reference product 2 (4152 U / g as polygalacturonase activity as measured above based on 1 g of tea leaves) were dissolved and dissolved. Enzyme treatment was performed at 0 ° C. for 8 hours.
After the enzyme treatment, the mixture was sterilized at 90 ° C. for 10 minutes, cooled to 30 ° C., and the solid residue of tea leaves was removed with an exposed cloth. 2 Using a Nutsche filter pre-coated with 10 g of cellulose powder on filter paper (8 cm), suction filtration (decompression degree 13.33 KPa) was performed at a constant pressure to obtain 825 g of a clear extract (required filtration time 3 minutes 42) Seconds). This extract was concentrated under reduced pressure to obtain 165.3 g of a Bx48 ° concentrate. This concentrated liquid was sterilized by heating at 95 ° C. for 30 seconds, filled into a sealed container, and then rapidly cooled to room temperature to obtain a green tea extract of the present invention product 1.
Example 2
In Example 1, the addition amount of Reference Product 2 was changed to 4.8 g, but was changed to 2.4 g (2076 U / g as the polygalacturonase activity based on the above measurement with respect to 1 g of tea leaves). (The time required for filtration: 4 minutes and 25 seconds) The product 2 (149.1 g) of the present invention was obtained.
Example 3
In Example 1, the amount of addition of Reference Product 2 was changed to 4.8 g, but 1.2 g (10 g U / g as the polygalacturonase activity as measured above from 1 g of tea leaves) was exactly the same as Example 1. (The time required for filtration was 5 minutes and 52 seconds). A product 3 (138.5 g) of the present invention was obtained.
Example 4
Example 1 is the same as Example 1 except that cellulosin PE60 (5.0 g, 1030 U / g as the polygalacturonase activity by the above measurement per 1 g of tea leaves) is added instead of Reference product 2 (4.8 g). The same operation was carried out (required filtration time: 5 minutes 21 seconds) to obtain the product 4 of the present invention (146.3 g).
Example 5
In Example 1, in addition to Reference product 2 (4.8 g), Sumiteam C (cellulase derived from Trichoderma longibrachiatum manufactured by Shin Nippon Chemical Industry Co., Ltd .: 0.25 g) was completely added to Example 1 except that 0.25 g was added. The same operation was performed (required filtration time: 3 minutes 21 seconds) to obtain Product 5 (167.3 g) of the present invention.
Example 6
In Example 1, in addition to Reference product 2 (4.8 g), the same operation as in Example 1 was carried out except that 0.25 g of cellulosin T3 (cellulase derived from Trichoderma reesei manufactured by Hibiai Co., Ltd .: 2600 U / g) was further added. (The time required for filtration was 3 minutes and 32 seconds) to obtain the product 6 (165.4 g) of the present invention.
Reference example 3
150 g of Sumiteam MC (manufactured by Shin Nippon Chemical Co., Ltd.) (polygalacturonase activity by the above measurement: 1690 U / g) was dissolved and washed in 1500 g of ion-exchanged water, and the precipitate was removed by centrifugation (4,500 × g, 5 minutes). The collected product was further freeze-dried to obtain Reference Product 3 (9.8 g, polygalacturonase activity by measurement described above: 20770 U / g).
Example 7
In Example 1, in place of Reference Product 2 (4.8 g), Reference Product 3 is added with 4.9 g (1018 U / g as a polygalacturonase activity based on the above measurement for 1 g of tea leaves). The completely same operation was performed (required filtration time: 4 minutes 49 seconds) to obtain the product 7 (153.2) of the present invention.
Reference example 4
100 g of sucrase N (manufactured by Mitsubishi Chemical Foods) (polygalacturonase activity by the above measurement: 4550 U / g) was dissolved in 1000 g of ion-exchanged water, and Vivaflow (registered trademark) 50VF05P2 (fraction molecular weight 30,000: manufactured by Sartorius) ), And 25 ml of the non-passed part was collected and freeze-dried to obtain Reference Product 4 (10.0 g, polygalacturonase activity by the above measurement: 32,000 U / g). .
Example 8
Example 1 is exactly the same as Example 1 except that 5.0 g of reference product 4 (1600 U / g as polygalacturonase activity as measured above per 1 g of tea leaves) is added instead of reference product 2 (4.8 g). The same operation was performed (required filtration time: 4 minutes 16 seconds) to obtain product 8 (155.4 g) of the present invention.
Comparative Example 1
In Example 1, except that no enzyme was used, the same operation as in Example 1 was performed (filtering time 10 minutes 25 seconds) to obtain Comparative Product 1 (66.8 g).
Comparative Example 2
In Example 1, except for not using the reference product 2 (4.8 g), the same operation as in Example 1 was performed (filtering time 9 minutes 57 seconds) to obtain a comparative product 2 (72.9 g).
Comparative Examples 3-5
In Example 1, instead of the reference product 2 (4.8 g), 2.0 g of Sumiteam AP2 (manufactured by Shin Nippon Chemical Industry Co., Ltd.) (248 U / g as a polygalacturonase activity by the above measurement with respect to 1 g of tea leaves) ), Sumiteam MC (manufactured by Shinnippon Kagaku Kogyo Co., Ltd.) 2.0 g (33.8 U / g as polygalacturonase activity based on 1 g of tea leaves as described above), Sucrase N (manufactured by Mitsubishi Chemical Foods) 2.0 g ( Except that the polygalacturonase activity as measured by the above measurement was 91 U / g per 1 g of tea leaves, the same operations as in Example 1 were performed to obtain comparative products 3 to 5 (filtering time and yield other than It is shown in the following Table 1 together with the measured value).
Comparative Examples 6 to 8 (Examples in which a polygalacturonase activity for 1 g of tea leaves was set to 800 U or more by using a large amount of commercially available pectinase)
In Example 1, instead of the reference product 2 (4.8 g), Sumiteam AP2 (manufactured by Shinnippon Kagaku Kogyo Co., Ltd.) 8.0 g (for 1 g of tea leaves, 992 U / g as the polygalacturonase activity as measured above) ), Sumiteam MC (manufactured by Shinnippon Kagaku Kogyo Co., Ltd.) 50.0 g (based on 1 g of tea leaves, 845 U / g as polygalacturonase activity as measured above), Sucrase N (manufactured by Mitsubishi Chemical Foods) 20 g (based on 1 g of tea leaves) Except that the polygalacturonase activity was determined to be 910 U / g by the above measurement, the same operations as in Example 1 were carried out to obtain comparative products 6 to 8 (filtering time and yield, together with other measured values) (Shown in Table 1).
Component Analysis Inventive products 1 to 8 and comparative products 1 to 8 were measured for tannin, amino acid and galacturonic acid concentrations (% is based on mass).
Measurement method Amino acid: Amino acid automatic analyzer Tannin: Iron tartrate method Galacturonic acid: High-performance liquid chromatography (HPLC) method Yield from green tea raw materials of the present invention products 1 to 8 and comparative products 1 to 8 and measured values (concentrations) ) And filter station time are shown in Table 1 below.

As shown in Table 1, the present invention products 1 to 8 and comparative products extracted by adding tea raw materials to protease, tannase and 1 g of tea leaves in an amount such that the polygalacturonase activity is 800 U or more. 6 to 8 were extracted by adding less than 800 U of polygalacturonase to Comparative Product 1 that did not use any enzyme, Comparative Product 2 that was extracted by adding protease and tannase, and 1 g of protease, tannase, and tea leaves As compared with any of Comparative products 3 to 5, it is clear that the filtration time is significantly shortened and the workability is remarkably improved.
In addition, the shortening of the filtration time is a difference in minute units in the small amount of preparation, and is not a big difference. Generally, in the industrial production of extracts, the filtration step is a step of limiting the work time of the whole process. Yes, when industrial mass production (several to several tens of tons) is carried out, it is expected to be a significant improvement.
In addition, as shown in Table 1, the comparative products 2 to 8 extracted by adding protease and tannase and the products of the present invention 1 to 8 were compared with the comparative product 1 that did not use any enzyme. Even amino acids have increased significantly.
Inventive products 1 to 8 and comparative products 6 to 8 extracted by adding 800 U or more of polygalacturonase to 1 g of tea leaf, protease, tannase, and green tea raw material are prepared by adding only protease and tannase to the green tea raw material. Compared with the extracted comparative product 2, the yield of the extract (Bx48 °) was increased to about twice, and the extract was obtained in a very high yield. In addition to the enzyme used in the product 1 of the present invention, the extract yield was further increased in the product 5 of the present invention using the cellulase derived from Trichoderma longibrachiatum and the product 6 of the present invention using the cellulase derived from Trichoderma reesei.
The products 2 and 3 of the present invention are obtained by reducing the amount of polygalacturonase used in the product 1 of the present invention, and the yield of the extract (Bx48 °) is slightly less than that of the product 1 of the present invention. Compared with comparative products 2-5, the amount increased by about 1.3 to about 2 times, and it can be seen that the yield of soluble solids from tea materials is greatly increased by the method of the present invention.
Comparative product 1 which does not use any enzyme contains almost no galacturonic acid, and comparative product 2 in which only protease and tannase are allowed to act on the green tea raw material contains only about 0.06% by mass of galacturonic acid. However, Comparative Products 3 to 8 and Invention Products 1 to 8 extracted by adding pectinase contained galacturonic acid in an amount of 0.16% to 0.92% by mass. In particular, it has been found that the galacturonic acid concentration increases as the added polygalacturonase activity unit increases. The products 1 to 8 of the present invention extracted by adding 800 U or more of polygalacturonase to 1 g of tea leaves have a particularly high concentration of galacturonic acid in the extract of 0.66 to 0.94% by mass. It was.
On the other hand, the inventive products 1 to 8 had slightly lower amino acid concentrations and tannin concentrations than the comparative products 3 to 5. However, this is considered to be due to the relative decrease in the amino acid concentration and the tannin concentration due to an increase in the degradation component of the cell wall.
In addition, a tea product was extracted by adding an enzyme preparation of less than 20000 U / g as a protease, tannase, and polygalacturonase activity to 1 g of tea leaves in an amount of 800 U or more as a polygalacturonase activity. 6-8, although the solid content yield is large, the concentrations of amino acids, tannins, and galacturonic acids are relatively low compared to the products 1-8 of the present invention, and the excipients in the enzyme preparations in tea extracts It seems that the component derived from etc. has been contained.
Therefore, Table 2 below shows the soluble solids yield and the yield of each component (calculated from Table 1) from the green tea raw materials of the present invention products 1-8 and comparative products 1-8.

As shown in Table 2, compared with Comparative Product 1 which does not use any enzyme, Comparative Products 2-8 and Invention Products 1-8 extracted by adding protease and tannase have a higher amino acid yield from tea leaves. It has increased 4 to 5 times. In addition, in the products 1 to 8 of the present invention and comparative products 6 to 8 extracted by adding 800 U or more of polygalacturonase to 1 g of tea leaves in addition to protease and tannase, less than 800 U for 1 g of protease, tannase and tea leaves Compared with comparative products 3-5 extracted by adding polygalacturonase, the amino acid yield from tea leaves is about 20% higher.
As for the tannin yield from tea leaves, the products 1 to 8 of the present invention and the comparative products 3 to 8 extracted by adding polygalacturonase in addition to protease and tannase are accompanied by an increase in the solid content yield. Increased. In particular, in the products 1 to 8 of the present invention and the comparative products 6 to 8 extracted by adding 800 U or more of polygalacturonase to 1 g of tea leaves in addition to protease and tannase, the tannin yield from the tea leaves is the tea leaf mass. Compared with Comparative Product 1 that does not use any enzyme and Comparative Product 2 that uses protease and tannase, the yield is about 20% higher.
The products 1 to 8 of the present invention and the comparative products 6 to 8 have a galacturonic acid yield from tea leaves of about 0.80% to 1.54%, indicating that a large amount of galacturonic acid is produced.
On the other hand, the comparative products 6 to 8 use polygalacturonase activity units of the same level as the products of the present invention 3, 4 and 7, and the galacturonic acid yield is the same, but the solids yield is More than the products 3, 4 and 7 of the present invention, in particular, there were many comparative products 7 and then comparative products 8 with a large absolute amount of the enzyme preparation added. From this, it is expected that the comparative products 6 to 8 contain a large amount of components derived from excipients and the like in the enzyme preparation.
Sensory evaluation After the inventive products 1 to 8 and comparative products 1 to 8 were diluted 160 times (Bx 0.3 °) with ion-exchanged water, sensory evaluation was performed by 10 well-trained panelists. The evaluation method is very good: 10 points, good: 8 points, slightly good: 6 points, slightly bad: 4 points, bad: 2 points, very bad: 0 points for bitterness, sweetness, umami, and balance, respectively. Sensory evaluation was performed and comments were made. The average points and average contents of comments are shown in Table 3 below.

As shown in Table 3, Comparative Product 1 that does not use any enzyme has an evaluation that green tea has a weak umami taste, sweet taste, and a strong bitter taste, and has any bitter taste, sweet taste, umami taste, or balance. Even the evaluation was low. In addition, comparative product 2 extracted by adding only protease and tannase to green tea raw material has a stronger taste of green tea and a bitter astringency than comparative product 1, but it is still quite strong and has a poor sweetness. The evaluation was somewhat higher than that of Comparative Product 1 for bitter astringency, sweetness, umami, and balance.
On the other hand, in addition to protease and tannase, an enzyme preparation having a polygalacturonase activity of 20000 U / g or more was added to 1 g of tea leaves in an amount such that the polygalacturonase activity was 800 U or more and extracted. The products 1 to 8 of the present invention have a strong taste, sweetness, and richness of green tea, a bitter and astringent taste, a well-balanced overall flavor, and taste like a high-quality matcha tea. there were.
On the other hand, comparative products 3 to 5 extracted by adding polygalacturonase of less than 800 U to 1 g of tea leaves in addition to protease and tannase have a slight bitter taste, although the taste and sweetness of green tea are felt to some extent. The balance was poor and the evaluation was inferior compared with the products 1 to 8 of the present invention.
Further, in addition to protease and tannase, an enzyme preparation having a polygalacturonase activity of less than 20000 U / g was added to 1 g of tea leaves in an amount of 800 U or more as a polygalacturonase activity, and extracted from comparative products 6 to 6 No. 8, green tea has a certain umami and sweet taste, but has a slightly different sweetness and miscellaneous taste compared to tea. In particular, Comparative Product 7 and Comparative Product 8 with a large absolute amount of the added enzyme preparation are , The sweetness and miscellaneous taste different from tea were felt strongly, the balance was bad, and the flavor was bad.
Ratio between ingredients Galacturonic acid has a mellow and refreshing acidity that makes you imagine a high-quality tea such as matcha tea, so it can be used for masking bitterness, masking off-flavors, adding a body sensation, etc. Therefore, it is presumed that an increase in galacturonic acid is one of the important factors for the sweetness, kokumi, umami and the like of the tea extracts obtained according to the present invention. That is, galacturonic acid exerts a masking effect in addition to the umami and sweetness of amino acids that are originally contained in teas and amino acids that are decomposed by protease treatment, masks the bitter and astringent taste of catechin, and is further produced by tannase treatment. It is presumed that the taste is improved by masking the acidity and egumi of gallic acid.
From the results shown in Tables 1 to 3, it was considered that the product of the present invention contained a relatively large amount of galacturonic acid as compared with other components. Therefore, the products of the present invention 1 to 8 and the comparative product 1 to 8: (a) galacturonic acid amount (mass) based on the total solid content of tea extract (Bx conversion), (b) galacturonic acid / tannin mass ratio, (c) galacturonic acid / amino acid mass ratio Calculated. The results are shown in Table 4 below.

As shown in Table 4, the products 1 to 8 of the present invention, which were very highly evaluated in terms of flavor, were (a) the content (mass) of galacturonic acid based on the total solid content (Bx conversion) of the tea extract. 1.3 to 2.0%, (b) the mass ratio of galacturonic acid / tannin is 0.07 to 0.12, and (c) the mass ratio of galacturonic acid / amino acid is in the range of 0.19 to 0.30. there were.
On the other hand, comparative products 1 to 5 have a content (mass) of galacturonic acid of less than 0.8% based on the total solid content (Bx conversion) of (a) tea extracts, and (b) galacturonic acid / tannin The mass ratio of (c) galacturonic acid / amino acid was less than 0.08.
Comparative products 6 to 8 have a content (mass) of galacturonic acid of 0.78 to 1.1% based on the total solid content (Bx conversion) of (a) tea extracts, and (b) galacturonic acid / The mass ratio of tannin was 0.059 to 0.07, and the mass ratio of (c) lacturonic acid / amino acid was 0.164 to 0.186, both of which were slightly lower than the products 1 to 8 of the present invention.
Therefore, it is presumed that the sweetness, richness, umami, etc. of the tea extracts obtained by the present invention were brought about by these differences.
In addition, as a numerical range, from the above examples, (a) the milk content (mass) of galacturonic acid based on the total solid content (converted to Bx) of tea extract is 1.1 to 5%. (B) the mass ratio of galacturonic acid / tannin is 0.04 to 0.8, and (c) the mass ratio of galacturonic acid / amino acid is 0.08 to 0.8; preferably (a) teas The content (mass) of galacturonic acid based on the total solid content (Bx conversion) of the extract is 1.2 to 4%, and (b) the mass ratio of galacturonic acid / tannin is 0.06 to 0.4. And (c) the mass ratio of galacturonic acid / amino acid is 0.14 to 0.6; more preferably, (a) the content of galacturonic acid based on the total solid content (converted to Bx) of the tea extract The amount (mass) is 1.3 to 3%, and (b) galacturonic acid / tannin mass ratio Is 0.07 to 0.2 and the mass ratio of (c) galacturonic acid / amino acid is 0.19 to 0.4, it is considered that the taste of the present invention is brought about.

Claims

Comprising at least tannin, amino acid and galacturonic acid,
(A) containing 1.1 to 5% by mass of galacturonic acid, based on the total solid content (Bx conversion) of the tea extract,
(B) the mass ratio of galacturonic acid / tannin is 0.04 to 0.8, and
(C) A tea extract having a galacturonic acid / amino acid mass ratio of 0.08 to 0.8.
The tea extract according to claim 1, wherein the galacturonic acid in the tea extract is produced by enzymatic decomposition of the tea raw material.
(A) containing 1.2 to 4% by mass of galacturonic acid based on the total solid content (converted to Bx) of the tea extract,
(B) the mass ratio of galacturonic acid / tannin is 0.06 to 0.4, and
(C) The tea extract according to claim 1, wherein the mass ratio of galacturonic acid / amino acid is 0.14 to 0.6.
(A) containing 1.3 to 3% by mass of galacturonic acid based on the total solid content of the tea extract (converted to Bx),
(B) the mass ratio of galacturonic acid / tannin is 0.07 to 0.2, and
(C) The tea extract according to claim 1, wherein the mass ratio of galacturonic acid / amino acid is 0.19 to 0.4.
A tea beverage containing the tea extract according to any one of claims 1 to 4.