JP2006199891A - Method for producing chamomilla extract - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for producing effectively a perfume ingredient, and anti-oxidant ingredient and other active ingredients from chamomilla. <P>SOLUTION: The method comprises acting glycosidase, protease and/or pectinase on chamomilla, so that a chamomilla extract being enhanced in an aromatic effect and an anti-oxidant activity can be produced. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an enzyme treatment method for producing a chamomile extract with enhanced aroma and enhanced antioxidant power.

  Chamomile has long been used for purposes such as sweating, lower fever, analgesia, anti-inflammation, and healthy stomach. There is also a demand for herbs because it has a good scent. For example, processed foods are increasingly used as beverages, confectionery candy, cookies, cakes, jelly, and other cosmetics and toiletries. Yes.

  Further, attention has been paid to the antioxidant power of flavonoids contained in chamomile, and proposals for application to foods, cosmetics and the like have also been made (see Patent Documents 1, 2, and 3).

As a method for extracting an active ingredient from chamomile, for example, a method of extracting chamomile flowers with aqueous C ₁ -C ₄ -alkanol having an alkanol content of 40 to 100% (see Patent Document 4), medicinal products such as chamomile A method of enzymatic treatment of a freeze-pulverized product of plant material with at least one degradation enzyme selected from the group consisting of protease, amylase, cellulase and pectinase in the presence of an aqueous medium (see Patent Document 5), chamomile stem and / or Or the method of extracting with the water-alcohol mixed solvent of a leaf (refer patent document 6 and 7) etc. is proposed.

JP-A-5-168429 JP 2004-75646 A JP 2004-262852 A Japanese Patent Publication No. 6-41414 JP-A-60-109526 Japanese Examined Patent Publication No. 6-104621 JP-A-5-112659

  However, the method of extracting active ingredients such as aroma components and antioxidant components from the above-described chamomile cannot be said to have sufficiently extracted these active ingredients, and the aroma components and antioxidant components contained in chamomile are not sufficient. It is insufficient from the viewpoint of effective use.

  Accordingly, an object of the present invention is to provide a method for efficiently extracting effective components such as aroma components and antioxidant components from chamomile.

  It is already known that when glycoside degrading enzymes are allowed to act on various plants, glycosides are decomposed and a new aroma is produced. Surprisingly, however, it was found that when a glycoside-degrading enzyme and protease and / or pectinase are allowed to act on chamomile, the odorous substance is remarkably increased and the antioxidant effect is further enhanced. The present invention has been completed.

  Thus, the present invention provides a method for producing a chamomile extract with enhanced aroma and enhanced antioxidant power, characterized by allowing a glycoside-degrading enzyme and protease and / or pectinase to act on chamomile. It is.

  According to the present invention, it is possible to provide a chamomile extract in which an aromatic substance several tens times as much as a conventional chamomile extract is obtained and the antioxidant effect is remarkably enhanced. For example, by adding the chamomile extract obtained by the method of the present invention to a food or drink, it provides a food or drink having a strong aroma and enhanced antioxidant properties compared to the case where a chamomile extract obtained by a conventional method is added. can do. Furthermore, for example, by blending in cosmetics and toiletries, a product with a strong fragrance can be provided, clothing can be protected, and oxidative conditions can be relaxed for the human body.

  Hereinafter, the present invention will be described in more detail.

  The chamomile that can be used as a raw material in the method of the present invention is a plant of the genus Compositae and genus Matricaria, Germania chamomill L., and other species of the same species, Camomile ( M. inodora L., M. matrixoides (Less.) Porter, M. tetragonosperma (Fr. Schm.) Hara et Kitam., Etc., and Roman camomiles (Anthemis noil). it can. In the present invention, these chamomile flowers, petals, stems, leaves, rhizomes, seeds, whole plants can be used, but preferably petals are used. The chamomile used in the present invention can be used either raw or dried, but it is preferable to use a dried product for ease of processing.

  As described above, the present invention is characterized in that the aroma is enhanced and the antioxidant power is enhanced by allowing glycoside degrading enzyme and protease and / or pectinase to act on these chamomiles.

    Specific examples of β-glucosidase used for glycoside enzyme treatment include, for example, β-glucosidase-producing bacteria belonging to the genus Aspergillus, Penicillium, Rhizopus, Pseudomonas, Pichia, etc., wheat bran, rice bran, etc. Examples thereof include solid culture medium or liquid culture medium in a solid nutrient medium or liquid nutrient medium according to a conventional method, and a culture obtained or a processed product thereof purified by a conventional method. As β-glucosidase, those obtained by purification from plants such as vanilla beans and fresh tea leaves can also be used, and further include almond-derived emulsine commercially available from Sigma-Aldrich, or β-glucosidase. Those separated from enzyme preparation cellulase A (manufactured by Amano Enzyme), cellulase T (manufactured by Amano Enzyme) and the like can also be used. As β-xylosidase, for example, β-xylosidase-producing bacteria belonging to the genus Penicillium, Aspergillus, Rhizopus, Mucor, etc. are solid or liquid cultured in a solid nutrient medium or liquid nutrient medium such as wheat bran or rice bran according to a conventional method. In addition, the culture obtained or the processed product obtained by purifying by a conventional method can be used. In addition, those derived from Aspergillus niger commercially available from Sigma-Aldrich, or those isolated from the enzyme preparation Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.) containing β-xylosidase can also be used. β-primeverosidase, for example, β-primeverosidase producing bacteria belonging to the genus Cellulomonas genus, Penicillium genus, Aspergillus genus and the like in solid or liquid culture in a solid medium or liquid medium such as wheat bran, rice bran, etc. The obtained culture product or a treated product thereof can be exemplified by those purified by a conventional method, and those obtained by separation and purification from plants such as fresh tea leaves can also be used. The amount of these glycoside-degrading enzymes varies depending on the titer and the like and cannot be generally specified, but can usually be exemplified in the range of 0.001 to 10 U / g based on the weight of the chamomile raw material.

  The protease is not particularly limited, and can be one or a combination of two or more proteases derived from animals and plants or microorganisms. For example, protease A, protease M, protease P, equinezyme, peptidase R, neurolase A , Newase F (Amanoenzyme-derived protease from Koji mold); Sumiteam AP, Sumiteam LP, Sumiteam MP, Sumiteam FP, Sumiteam LPL (Since Nippon Koji Co., Ltd., Koji mold-derived protease); Protin FN (Yamato Kasei-derived protease from Kasei Co., Ltd .; Denapsin 2P, Denateam AP, XP-415 (and above, Neisseria mutexes-derived protease from Nagase ChemteX); Orientase 20A, Orientase ONS, Tetrase S (above, HI Morocin F, PD enzyme, IP enzyme, AO-protease (above, Kikkoman's Neisseria gonorrhoeae protease); Sakanase (Kanken Pharma's Neisseria gonorrhoeae-derived protease); Punchase YP-SS, Pantidase NP-2, Pandidase P (above, Koji mold-derived protease manufactured by Yakult Pharmaceutical Co., Ltd.); Flavorzyme (Novozymes Koji mold-derived protease); Coclase SS, Coclase P (Sankyo Lifetech, Koji mold-derived protease) ); VERON PS, COROLASE PN-L (above, a protease derived from Aspergillus manufactured by AB Enzyme); P, Deskin, Depilace, Protin A, Samoaase (bacteria-derived protease manufactured by Daiwa Kasei Co., Ltd.); Biolase XL-416F, Biopase SP-4FG, Biopase SP-15FG (Bacteria-derived protease manufactured by Nagase ChemteX Corporation) ); Orientase 90N, nucleicin, orientase 10NL, orientase 22BF (above, bacteria-derived protease manufactured by HIBI); aloase AP-10 (bacteria-derived protease manufactured by Yakult Pharmaceutical Co., Ltd.); protamex, neutrase, Alcalase (above, bacterial protease from Novozymes); COROLASE N, COROLASE 7089, VERON W, VERON P (above, bacterial protease from AB Enzyme); Entiro NBS (Bacteria-derived protease manufactured by Nitto Kasei Kogyo Co., Ltd.); Alkaline protease GL440, Purefect 4000L, Protease 899, Protex 6L (Bacteria-derived protease manufactured by Genecon Kyowa); Actinase AS, Actinase AF (above, Kaken Pharma) Actinomycetes-derived protease); Tacinase (Protein-derived protease manufactured by Genecon Kyowa); Papain W-40 (Amanoenzyme plant-derived protease); Food-purified papain (Nagase Chemtex plant-derived protease) ); Other animal-derived pepsin, trypsin, and the like. The amount of these proteases to be used varies depending on the titer and the like and cannot be generally specified, but can usually be exemplified in the range of 0.01 to 100 U / g based on the weight of the chamomile raw material.

  Pectinases are known to be contained in bacteria, molds, yeasts, higher plants, snails, etc. In the present invention, pectinases obtained from organisms including these can be widely used. A commercially available pectinase preparation may also be used. Examples of commercially available pectinase preparations include sucrase (manufactured by Sankyo Lifetech), pectinex ultra SP-L (manufactured by Novozymes), mecerase (manufactured by Meiji Seika Co., Ltd.), ultrazyme (manufactured by Novozymes), and newase F ( Examples include Amano Enzyme). The amount of use of these pectinases varies depending on the titer and the like and cannot be generally specified, but can usually be exemplified within the range of 10 to 2000 U / g based on the weight of the chamomile raw material.

  The above-described treatment of chamomile with the enzyme is a method known per se, for example, the Patent Office Gazette Well-known and conventional technology collection (fragrance) Part II Food Fragrance (issued 2000.1.14) Microorganism / Enzyme Flavor (P46-P57) It can carry out according to the method as described in such publications. An embodiment of the present invention is exemplified as follows. 8 to 30 parts by weight of water is added to 1 part by weight of the chamomile raw material, sterilized at about 60 to about 121 ° C. for about 2 seconds to about 20 minutes, and then cooled, and the above glycoside-degrading enzyme and protease and / or Pectinase is added and the enzyme treatment is performed at about 20 to about 60 ° C. for about 30 minutes to about 24 hours. After the enzyme treatment, the enzyme is inactivated by heating at about 60 to about 121 ° C. for about 2 seconds to about 20 minutes, cooled, and then clarified by separating the chamomile raw material by appropriate separation means such as centrifugation and filter paper filtration. Can be obtained. If desired, the obtained chamomile extract can be in the form of a concentrated solution by an appropriate concentration means such as vacuum concentration, reverse osmosis membrane concentration, and freeze concentration. Furthermore, the obtained chamomile extract can then be in any form such as a paste or powder, if desired.

  The chamomile extract of the present invention thus obtained can be blended in beverages such as soft drinks, carbonated drinks, milk drinks and functional drinks; and foods and drinks such as sweets such as candies, cookies, cakes and jellies. . Furthermore, for example, shampoos, hair creams, other hair cosmetic bases; oscillating lipsticks, other cosmetic bases and cosmetic detergent bases; laundry detergents, disinfecting detergents, deodorant detergents And other various health and sanitary detergents; various health and hygiene materials such as toothpaste, tissue, and toilet paper; and pharmaceuticals.

  Hereinafter, the present invention will be described more specifically with reference to Examples and Comparative Examples.

Example 1
180 g of soft water was added to 12 g of Roman chamomile (Nippon Green Tea Center Co., Ltd.), and this was heated to 80 ° C. with stirring. This was cooled to 40 ° C., 5 units of emulsin (manufactured by Sigma Aldrich), 5 units of β-xylosidase (manufactured by Sigma Aldrich), 0.02 g of protease A (manufactured by Amano Enzyme) and 0 of sucrase N (manufactured by Sankyo) 0.02 g was added, stirring was stopped, the mixture was allowed to stand at 40 ° C. for 16 hours to react, then heated again with stirring, deactivated by heating at 75 ° C. for 15 minutes, and cooled to 40 ° C. The reaction solution was filtered with a bare cloth, further centrifuged (800 g × 20 minutes), and then filtered with a filter paper precoated with diatomaceous earth to obtain 155 g of chamomile extract (Bx3.0: Invention product 1).

Comparative Example 1
In Example 1, except for not using all enzymes, it processed like Example 1 and obtained 120 g of chamomile extracts (Bx1.5: comparative product 1).

Example 2
In Example 1, except that Roman chamomile was changed to German chamomile (manufactured by Nippon Green Tea Center Co., Ltd.), the same treatment as in Example 1 was carried out to obtain 135 g of chamomile extract (Bx3.8: Invention product 2).

Comparative Example 2
In Example 2, chamomile extract 115g (Bx2.0: Comparative product 2) was obtained in the same manner as in Example 2 except that no enzyme was used.

Example 3
In Example 2, 130 g of chamomile extract (Bx3.6: product 3 of the present invention) was obtained in the same manner as in Example 2 except that protease A was not used.

Example 4
In Example 2, a chamomile extract 125g (Bx3.4: product 4 of the present invention) was obtained in the same manner as in Example 2 except that sucrase N was not used.
(sensory evaluation)
About each chamomile extract of this invention products 1-4 and comparative products 1-2, it diluted 10 times with ion-exchange water, and sensory evaluation was performed by ten panelists who were well trained in the flavor. The results are shown in Table 1.

(Aroma analysis)
About the chamomile extract of this invention product 1 and comparative product 1, and this invention product 2 and comparative product 2, aroma analysis was performed by the dynamic head space method shown below, and the difference in aroma was compared.

Aroma analysis (dynamic headspace method)
Collect 5 g of each of the chamomile extracts of the present invention products 1 and 2 and comparative products 1 and 2 in a 500 mL 2-diameter flask, and keep the sample at 40 ° C. while keeping the sample at 40 ° C. Nitrogen gas was blown at / min., A cooling pipe and an adsorbent (TENAX TA) were connected to the other port, and the fragrance expelled from the sample was adsorbed for 30 minutes. The adsorbent which adsorb | sucked a fragrance carried out the gas chromatographic analysis on the following conditions by carrying out the heat | fever desorption of a fragrance | flavor component by Thermo Dessection System made from GERSTEL, and the obtained gas chromatogram is shown in FIG.

Gas chromatography analysis conditions
Model: Hewlett Packard HP-6890
Column: Fused Silica Capillary
OV101 60m x 0.25mm
Column temperature: 70 to 220 ° C. (3 ° C./min)
Injection temperature: 250 ° C
Detector temperature: 250 ° C
Carrier gas: N2 1.8 kg / cm ²
(Gas chromatography analysis results)
Compared to the total integrated value 25045.2 of the sample of the comparative product 1, the total integrated value of the sample of the product 1 of the present invention was 37315.5, and the amount of fragrance of the product 1 of the present invention was larger. That is, the total integrated value increased 1.5 times by treating the Roman chamomile with the enzyme. The components in which the integrated value of the product 1 of the present invention was remarkably increased compared to the comparative product 1 were isobutyraldehyde, methyl vinyl ketone, diacetyl, and 2-methyl-3-buten-2-ol.

  Further, the total integrated value of the sample of the present invention 2 is 9927.6 compared to the total integrated value 2711.0 of the sample of the comparative product 2, and the total integrated value of 3.7 is obtained by enzymatic treatment of German chamomile. Doubled. Ingredients in which the integrated value of the product of the present invention 2 is significantly increased compared to the comparative product 2 are 2-methylbutanol, isoamyl alcohol, 3-methyl-3-butenol, 2-heptanol, prenyl alcohol, trans-3-hexenol , Alcohols such as cis-3-hexenol, 2,5,5-trimethyl-3,6-heptadien-2-ol, trans-2-hexenol, heptanol, octanol, cis-3-nonenor, methyl heptenone, artemisia ketone, etc. Ketones.

(Antioxidant)
About each chamomile extract of this invention product 1 and the comparative product 1, and this invention product 2 and the comparative product 2, the antioxidant property of each chamomile extract was measured by the hypoxanthine-xanthine oxidase system antioxidant ability measuring method shown below. .
Hypoxanthine-xanthine oxidase system antioxidant capacity
[1] Reagent (a) Substrate (for pump injection)
A solution obtained by dissolving 24.5 mg of hypoxanthine in 250 mL of 100 mM potassium dihydrogen phosphate buffer (containing 0.05 mM EDTA, pH 7.5: hereinafter simply referred to as buffer) was used.
(B) Enzyme solution 994 μL of buffer solution was added to 6 μL of xanthine oxidase (Sigma-4 Aldrich X-4500; 25 U) to adjust to 0.1 U / mL.
(C) Coloring reagent AB-2950MPEC (2-methyl-p-methoxyphenylimidazopyrazinone) was diluted 10 times with ion-exchanged water and used.

[2] Sample solution (d) Positive blank 180 μL of buffer solution, 60 μL of enzyme solution and 10 μL of luminescent reagent were mixed.
(E) Sample measurement solution 170 μL of buffer solution, 60 μL of enzyme solution, 10 μL of luminescent reagent, and 10 μL of sample solution (diluted 10 times or 300 times) were mixed.
(F) Negative blank 230 μL of buffer solution, 10 μL of luminescent reagent and 10 μL of sample solution (10-fold dilution or 300-fold dilution) were mixed.
(G) SOD solution 1.0 g of superoxide dismutase (S-2515; 3000 U, manufactured by Sigma-Aldrich) was dissolved in pure water to make 3 mL, and a 1,000 U / mL solution was prepared. This solution was diluted with pure water to make 0.1 U / mL, 1 U / mL, 10 U / mL, and 100 U / mL.

[3] Measurement Using a luminescence sensor JNR2 (manufactured by Ato Co., Ltd.), d, e, f, and g were measured under the following conditions.
Measurement conditions Substrate dispensing volume: 50 μL
Measurement mode: Instant emission Measurement time: Integrated light emission for 20 seconds Measurement temperature: 20 ° C
In addition, it was confirmed that the negative blank did not emit each sample itself.

[4] Calculation Luminescence inhibition rate (%) = [1- (e) / (d)] × 100
(D) Integrated luminescence value of positive blank (e) Integrated luminescence value of sample measurement solution The luminescence inhibition rate of the SOD diluted solution was taken as a calibration curve, and the SOD equivalent amount of each sample was calculated, and the results are shown in FIG.

  As is apparent from the results of FIG. 3, the products 1 and 2 of the present invention treated with the enzyme have enhanced antioxidant properties (SOD-like activity) compared to the products 1 and 2 which are not treated with the enzyme. It was.

It is a figure which shows the gas chromatogram which measured the chamomile extract of this invention product 1 and the comparative product 1 with the dynamic head space method. It is a figure which shows the gas chromatogram which measured the chamomile extract of this invention product 2 and the comparative product 2 by the dynamic head space method. It is a figure which shows the measured value which measured the antioxidant property of the chamomile extract of this invention products 1 and 2 and the comparative products 1 and 2 by the hypoxanthine-xanthine oxidase type antioxidant capacity measuring method.

Claims (3)

A method for producing a chamomile extract with enhanced aroma and enhanced antioxidant power, characterized by allowing a glycoside-degrading enzyme and protease and / or pectinase to act on chamomile. The production method according to claim 1, wherein the glycoside degrading enzyme is at least one selected from β-glucosidase, β-primeverosidase, and β-xylosidase. A chamomile extract obtained by the production method according to claim 1 or 2.

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