JPH0995672A - Readily water-soluble myricaceae plant extract - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject extract excellent in solubility in water and antioxidant effect, and usable as a water-soluble antioxidant for foods, medicines, cosmetics, etc., by making an enzyme act on a Myricaceae plant extract in the presence of a galactose residue transfer source. SOLUTION: (C) An enzyme having galactose residue transfer activity (e.g. β-D-galactoside galactohydrase) is made to act on (A) a Myricaceae plant extract having antioxidant effect (for example, obtained by the following process: a ground product of the bark, rhizome, twigs or leaves of Myrica rubra or gale is subjected to extraction with an organic solvent such as a 1-5C aliphatic alcohol or 3-5C carbonyl compound followed by removing water-soluble substances such as condensed type tannin, catechins and saccharides from the extracted product) in the presence of (B) a galactose residue transfer source (e.g. lactose, galactooligosaccharide) to effect transferring galactose residues to the component A.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an easily water-soluble extract of the bayberry family having an antioxidant effect. The water-soluble Sophoraceae plant extract obtained in the present invention can be used for producing foods, pharmaceuticals, quasi drugs, cosmetics or feeds.

[0002]

2. Description of the Related Art Products such as oils and fats or foods containing oils and fats, pharmaceuticals, quasi-drugs, cosmetics and feed are oxidized,
It is well known that it causes resin deterioration, off-flavor, coloration, discoloration, generation of toxic substances, or deterioration of nutritional value, resulting in deterioration of quality. Conventionally, synthetic antioxidants such as butylhydroxyanisole (hereinafter referred to as BHA) and butylhydroxytoluene (hereinafter referred to as BHT) have been used as antioxidants, and tocopherols, L- Ascorbic acid, sesamolin in sesame oil, caffeic acid derivative, melanoidin, amino acid, phytic acid, tea leaf extract, spice extract such as rosemary and sage, and the like alone or a mixture thereof is used.

[0003]

DISCLOSURE OF INVENTION Problems to be Solved by the Invention The inventors of the present invention have made a detailed study on the extract of the family Asaceae, and as a result, have found that the extract of the family Asaceae contains a substance having a strong antioxidant effect. In particular, it has been found that a substance having an excellent antioxidative effect which is excellent in thermal stability and which meets the demand of the industry can be obtained in the poorly water-soluble fraction. In order to effectively utilize this excellent function in the industry, we plan to modify the extract of the bayberry family that is sparingly water-soluble and difficult to handle, dissolve it in water and water-containing organic solvent systems, and stabilize it. It is an important subject to provide a method for preventing deterioration of quality of foods, pharmaceuticals, quasi drugs, cosmetics or feeds due to oxidation, as well as to provide antioxidants.

[0004]

In view of the above problems, the present inventors have retained the excellent antioxidant effect of the bayberry family extract while maintaining the solubility and stability in an aqueous system or a water-containing organic solvent system. The method for improving the sex was earnestly studied. As a result, in the coexistence of a bayberry plant extract and a galactose residue transfer source, by reacting an enzyme having a galactose residue transfer activity, a water soluble soluble bayberry in which a galactose residue was transferred to a bayberry family extract Invented a plant extract, which was used in foods, drugs, quasi drugs, cosmetics,
It has come to be provided as a method for preventing quality deterioration by using it as feed. The details will be described below.

The poorly water-soluble plant extract of the bayberry family, which is the antioxidant substance of the raw material used in the present invention, is a bayberry (Myrica rubra SIEB. Et ZUCC.) And / or a willow tree (Myrica gale L.) of the genus Bayberry family. Extract from. Prior to extracting the antioxidant from these bayberry plants, bark, rhizomes, branches or leaves are crushed using a crusher. The method for extracting the antioxidant is the method already filed by the applicant (Japanese Patent Laid-Open No. 15624/1993)
9, Japanese Patent Application No. 7-250902) can be used.
That is, a method of extracting an antioxidant substance or the like by a general means such as immersing the plant body in an organic solvent, and then removing tannin, condensed tannin, catechins, sugars and other water-soluble substances from the extract Alternatively, it is also a method of previously removing these water-soluble substances from the plant and then extracting the antioxidant substance with an organic solvent.

The extract obtained by any method is composed of a sparingly water-soluble extract of the bayberry family and can be a raw material used in the present invention. The organic solvent used for this extraction has 1 carbon
To 5 as the aliphatic alcohol organic solvent, methanol, ethanol, propanol, isopropanol,
Butanol, isobutanol, 2-butanol, pentanol, 1,2-propanediol, 1,3-propanediol,
Examples of other organic solvents include carbonyl compounds having 3 to 5 carbon atoms such as acetone, 2-butanone, 2-pentanone, and 3-pentanone, and water-soluble acid amides such as formamide and N, N-dimethylformamide, N, N-diethylformamide, N, N-
Examples of the water-soluble amine such as dimethylacetamide include pyridine, butylamine, and the like, and dimethylsulfoxide and the like, which may be used alone or in an appropriate combination. Further, water may be appropriately used in combination with the above organic solvent, if necessary.

In the method for extracting the plant with an organic solvent and then removing the water-soluble substance from the extract to obtain an antioxidant, the plant is dipped in the above-mentioned solvent by a general method. The solvent is removed from the extract obtained by the treatment by evaporation or other common means. Water is added to and mixed with the concentrated liquid and the concentrated dried product obtained by this operation, and water-soluble substances such as catechin, tannin, condensed tannin and sugar are transferred to the water phase side and removed to make it poorly water-soluble. To give the antioxidant as a solid. The obtained product is the sparingly water-soluble bayberry plant extract used in the present invention. In another method of extracting the antioxidant substance with an organic solvent after removing the water-soluble substance from the plant, the water-soluble substance is extracted by a general means such as immersing a crushed product of bark, rhizome, branch or leaf in water. Material is extracted and removed.

At this time, if necessary, the ground product may be subjected to a steaming treatment and then extracted. Further, one kind selected from sodium hydroxide, sodium carbonate, sodium hydrogen carbonate, potassium hydroxide, potassium carbonate, potassium hydrogen carbonate, ammonia gas, ammonia water, and ammonium hydrogen carbonate in order to more easily extract and remove the water-soluble substance. Alternatively, two or more kinds may be used to adjust the pH of the liquid at the time of extraction so as to fall within the range of 4 to 8. For the purpose of reducing the amount of solvent used in extraction with an organic solvent and facilitating the recovery and purification work of the solvent, the obtained pulverized product is cold-air dried, hot-air dried, freeze-dried or other You may dry by a method etc. Further, in order to improve extraction efficiency and workability, the crushed product may be molded into pellets or granules. The product subjected to such a treatment is a pulverized product of the bayberry family with water-soluble substances removed. Then, the poorly water-soluble antioxidant substance can be obtained by immersion in the above-mentioned organic solvent or extraction by another general method.
If necessary, it may be further purified by washing with water or hot water, purification by chromatography or liquid-liquid countercurrent extraction, or recrystallization from an organic solvent or a water-containing organic solvent. .

The bayberry plant extract obtained by such an operation contains myricitrin (myricetin 3-O-3 rhamnoside), which is a flavonol glycoside, and other compounds as active ingredients, and has a strong antioxidant effect. , But
It is poorly soluble in water and water-containing organic solvents, and is a substance that is difficult to handle in order to exert its excellent effects. The problem that this solubility is low and difficult to handle is that substances such as mylycitrin and other flavonol glycosides that are insoluble in the bayberry plant extract are transferred to a galactose residue of the present invention, which is easily soluble in bayberry. It was solved by using the family plant extract.

A water-soluble soluble bayberry plant extract of the present invention,
The galactose residue transfer source used in the present invention may be a substrate for an enzyme having a galactose residue transfer activity, and one or more molecules of the galactose residue can be transferred to a bayberry family extract, for example, lactose alone. , Lactose known method (Agric. Biol. Chem., 48: 3053-306).
Page 1 (1984)) galacto-oligosaccharides obtained by transferring galactose residues or commercially available galacto-oligosaccharides derived from lactose, galacto-oligosaccharides derived from beans such as soybean, and the like. 1
One kind or two or more kinds are adopted. The galactose residue source may be added in an amount of 1 to 80% by weight (hereinafter, indicated by%), preferably 10 to 70%, based on the whole reaction mixture.
More desirably, an amount of about 20 to 60% is advantageous.

The enzyme having a galactose residue transfer activity used in the present invention is preferably β-D-galactoside galactohydrase (EC 3.2.1.23, hereinafter referred to as β-galactosidase). For example, Bacillus circulans (Bacillus circulans), Bacillus genus such as Bacillus macerans (B. macerans), Lactobacillus bulgaricus, Lactobacillus lactis (L. lactis), Lactobacillus plantarum (L. plantarum), Escherichia Escherichia coli and other Aspergillus oryzae
e), Aspergillus genus such as A. niger, Kluyveromyces
lactis), Kliberomyces fragilis (K. fragilis)
Such as Kleberomyces spp., Streptococcus thermophilus and other Streptococcus spp., Helix pomatia, and other Helix spp., Penicillium chrysogenum (Penicilliu)
m crysogenum), Penicillium multicolor (P.multi)
color) etc. Penicillium genus, Saccharomyces fragilis (Saccharomyces fragilis) genus Saccharomyces,
Others originated from microorganisms such as conch (Chalonia
lampas) and other sources derived from shellfish, those derived from plants such as jack bean (Japanese name: jack bean, Canavalia ensiformis), or those originating from the liver of cattle or the small intestine of mammals. Yes, it can be used freely in this invention.

It is not always necessary to purify and use these enzymes, and a crude enzyme can usually achieve the purpose. In addition, a commercially available enzyme preparation (for example, product name B manufactured by Daiwa Kasei Co., Ltd.
IOLACTA G10 etc.) can also be used. In addition, instead of adding β-galactosidase to a culture solution containing a bayberry plant extract and a galactose source, a β-galactosidase-producing bacterium having a galactose residue transfer activity is inoculated, and a glycosyl transfer reaction is performed by a fermentation method. You can also do it. Furthermore, the reaction may be carried out using β-galactosidase or a β-galactosidase-producing microorganism immobilized in accordance with a conventional method. Even if these β-galactosidases originate from a single species of microorganism, plant or animal are used, two or more enzymes of different origins can be used in combination, and β-galactosidase producing bacteria It is also possible to inoculate more than one seed and perform the glycosyl transfer reaction by fermentation.

The amount of β-galactosidase used is not particularly limited. The amount of this enzyme used varies greatly depending on the source and the dosage form of the enzyme. For example, even when the same enzyme is used, the amount used varies greatly depending on whether it is used as an enzyme solution or immobilized. Therefore, since it cannot be determined unambiguously, for example, when BIOLACTA G-10 is used, an amount of about 10 to 2000 units / g substrate is usually advantageous. The enzyme unit will be described in detail in Examples. In addition, as an enzyme activator, Mn may be added if necessary.
Metal ions such as 2+, Mg2 + and Ca2 + may be used in combination with the enzyme. The addition amount thereof is usually small and is selected from the range of 1 to 500 ppm with respect to the reaction mixture system. The pH of the reaction system in this transfer reaction is preferably around the optimum pH of the enzyme used, and it is usually preferable to select it in the range of about 2-9. The temperature of this transfer reaction is preferably around the optimum temperature of the enzyme used, and it is usually preferable to select it from the range of 20 to 70 ° C.

Thus, the galactose residue-transferred bayberry plant extract of the present invention can be produced in a high yield by a simple operation, and the purpose of the production of the water-soluble bayberry plant extract of the present invention is to Can be reached. Furthermore, if desired, the reaction system is treated with an ion exchange resin or an ion exchange membrane, a resin having a porous polymer structure, silica gel,
It may be purified by an adsorption chromatographic treatment using aluminum oxide, cellulose, or the like as an adsorbent, a reverse phase partition chromatographic treatment using activated carbon, an alkylsilylated silica gel, an arylsilylated silica gel, or the like as an adsorbent.

A galactose residue-transferred bayberry family extract (water-soluble bayberry family plant extract), which is a modified bayberry family plant extract produced by the above method, constitutes a sparingly water soluble bayberry family plant extract. By transferring galactooligosaccharide residues to the sugar chain structure of flavonol glycosides, the solubility in water and water-containing organic solvents is improved, and not only it exerts its effect as an antioxidant but also water-soluble. It can also be used as a deodorant for sexuality, and when it contains an oligosaccharide chain portion, it is expected to be effective as a growth promoter for Bifidobacterium.

The water-soluble extract of the bayberry family of the present invention contains corn oil, rapeseed oil, cottonseed oil,
Soybean oil, safura oil, sunflower oil, sesame oil, wheat germ oil, olive oil, evening primrose oil, camellia oil, tea seed oil, avocado oil, castor oil, coffee oil, cashew nut oil, cacao bean oil, peanut oil, fish oil, palm oil, palm. Nuclear oil,
Animal and vegetable oils and fats such as lard, beef tallow, chicken and partially hydrogenated oils and fats of these animal and vegetable oils or fats, oleic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, eicopentaenoic acid, docosahexaenoic acid Not only can it be used for unsaturated fatty acids and esters thereof or unsaturated alcohols thereof, but also for processed oils and fats such as butter, margarine, shortening and dressing.

Further, it can be added to foods containing a large amount of fats and oils, for example, donuts, fried foods, fried foods, chocolates, instant ramen and the like. In addition, Japanese sweets such as okaki, senbei, okoshi, manju, and candy,
Cookies, biscuits, crackers, pies, sponge cakes, castella, donuts, waffles, puddings, butter cream, custard cream, cream puffs, chocolate, chocolate confectionery, caramel, candy, chewing gum, jelly, hot cakes, bread and other Western confectionery, potatoes Snack snacks such as chips, ice cream, ice lollies, frozen desserts such as sorbet, lactic acid bacteria drinks, milk drinks, fruit juice drinks, fruit juice-free drinks,
Fruit drinks, functional drinks, clear carbonated drinks, carbonated drinks containing fruit juice, soft drinks such as fruit-colored carbonated drinks, green tea, tea, instant coffee, cocoa, coffee drinks in cans, favorite drinks such as commercial coffee, fermented milk, processed milk,
Dairy products such as cheese, soybean processed foods such as tofu and soy milk,
Marmalade, jam, fruit syrup pickling, paste such as flower paste, peanut paste, fruit paste, pickles, ham, sausage, bacon, meat products such as dry sausage, beef jerky, fish ham, fish sausage, kamaboko, Fish paste products such as Chikuwa, Hanpen, and Tempura, various dried fish such as fish, squid, octopus, shellfish and their delicacies, various sections such as bonito, mackerel, and eel, sardines, sea urchin, salted squid, dried fish only. Smoked products such as salmon, seaweed, small fish, shellfish, sea bream, wild vegetables, mushrooms,
Tsukudani cooked with kelp, curry such as instant curry, retort curry, canned curry, miso, powdered miso, soy sauce, powdered soy sauce, moromi, fish sauce, sauce, ketchup, mayonnaise, solid bouillon, sesame oil, roasted meat It can be used for various seasonings such as sauce, curry roux, stew base, soup base, dashi base, etc., and also various foods and drinks such as various range foods and frozen foods containing fats and oils and their derivatives. , Can also be used for luxury items.

In addition, as pharmaceuticals, quasi-drugs and cosmetics, it is free for drinks, troches, liver oil drops, mouthwash, mouthwash, mouthwash tablets, toothpaste, sunscreen skin lotion, UV protection cream, lipstick, etc. As a feed, it can be added to various cat foods, dog foods, ornamental fish feeds, cultured fish feeds, and the like.

The easily water-soluble bayberry plant extract of the present invention can be dissolved in water or a suitable solvent such as ethanol, propylene glycol or glycerin, even if it is used as a powder depending on the shape of the target product. Or as an emulsion. The product of the present invention can also be used in combination with other antioxidants such as tocopherol, L-ascorbic acid, sodium erythorbate, BHA and BHT. Water-soluble soluble bayberry plant extract of the present invention, food, drug, quasi drug,
When used in cosmetics, feed, etc., the amount added is usually 0.0002 to 5% (weight, the same below), and preferably 0.002 to 2%. Examples of the present invention will be shown below.

[0020]

【Example】

Extraction Example 1 10 kg of methanol was added to 1 kg of a dried product of bayberry bark, and the mixture was extracted at about 60 ° C. for 5 hours and then filtered, and the residue was washed with 3 kg of methanol to obtain about 10 kg of a methanol extract.
After concentrating this extract, transfer it to another container, and vacuum degree 5mmH
g, and dried under reduced pressure at a bath temperature of 60 ° C. to obtain 0.25 kg of a yellow powder.
The obtained solid matter was pulverized, suspended in 5 L of water at room temperature and then filtered, and the remaining solid matter was washed with 5 L of water. Then, the solid content is dried under reduced pressure at a vacuum degree of 5 mmHg and a bath temperature of 80 ° C. and dried under reduced pressure to form a bayberry plant extract (referred to as extract 1) 0.13.
kg gained.

Extraction Example 2 10 kg of water was added to 1 kg of the dried product of bayberry bark, and the mixture was stirred and mixed at room temperature for 6 hours at 60 ° C., and the water-soluble substance was removed using a centrifuge to obtain a solid product. Was dried with warm air at 65 ° C. to obtain 780 g of crushed bark from which water-soluble substances were removed. To this dried product, 8 kg of ethanol was added, and the mixture was stirred and mixed at about 80 ° C. for 5 hours and then filtered, and the residue was washed with 3 kg of ethanol to obtain about 9 kg of an ethanol extract. The extract was concentrated and transferred to another container, and dried under reduced pressure at a vacuum degree of 5 mmHg and a bath temperature of 60 ° C. to obtain 0.21 kg of a bayberry plant extract (referred to as extract 2) consisting of yellow powder.

Extraction Example 3 800 g of ethanol was added to 100 g of dried ground willow bark bark, and the mixture was extracted at about 80 ° C. for 5 hours and filtered, and the residue was washed with 200 g of ethanol to obtain 800 g of an ethanol extract. After concentrating this extract solution, transfer it to another container, vacuum degree 5 mmHg,
After drying under reduced pressure at a bath temperature of 60 ° C., 20 g of a yellowish brown powder was obtained. The obtained solid was crushed, suspended in 200 ml of water at room temperature and filtered, and the remaining solid was washed with 100 ml of water. Then, the solid content is dried under reduced pressure at a vacuum degree of 5 mmHg and a bath temperature of 80 ° C. and dried under reduced pressure to form a yellow-brown solid matter of the bayberry family extract (referred to as extract 3).
Obtained 8.5 g.

Method for measuring β-galactosidase activity 0.05M phosphate buffer containing 0.1% p-nitrophenyl-β-D-galactoglycoside (pH is optimal for the enzyme.
Adjust to H) 0.2 ml, and add 0.1 ml of enzyme solution (2-5 mm units) appropriately diluted in 0.05M phosphate buffer to 40 ° C.
After reacting for 15 minutes, 2M of 1M sodium carbonate solution is added to the reaction solution.
The reaction is stopped by adding, and the absorbance at 420 nm is measured using a 1M sodium carbonate solution as a control using a spectrophotometer, and the enzyme unit is determined by the following formula. Enzyme unit = absorbance × 0.01 × 1 / enzyme concentration (g / ml) The molecular extinction coefficient of p-nitrophenol under these conditions is 15,000, and one enzyme unit obtained from the above formula is per minute. This corresponds to the amount of liberating 1 μM of p-nitrophenol from p-nitrophenyl-β-D-galactoside.

(Galactose Residue Transfer Method Analyzing Extracts of Pediculaceae) Measurements were carried out by high performance liquid chromatography under the following conditions. Column: Micro bonder pack C18, column diameter 4.6
mm, column length 250 mm Solvent: Methanol / acetonitrile / acetic acid / water = 7/1
/ 1/12 Flow rate: 1 ml / min Detector: UV / visible spectrophotometer Measurement wavelength: 350 nm Under these measurement conditions, the peak of the raw bayberry extract and the peak of the reaction product were completely separated.

Example 1 Lactose (250 g) was added to 100 ml of 0.1 M phosphate buffer (pH 7.0) to prepare 6
After heating and dissolving at 0 ° C, 100 ml of dimethyl sulfoxide solution containing 20 g of Extract 1 and 1 g of β-galactosidase (enzyme titer of 20,000) derived from Bacillus circulans manufactured by Daiwa Kasei Co., Ltd. were added to this solution at 60 ° C. It was stirred for 4 hours. After completion of the reaction, the mixture was diluted with 1 L of water to give a porous polymer 70 composed of a styrene-divinylbenzene copolymer.
The column filled with 0 ml was passed for 1 hour, and then 5 L of ion-exchanged water was passed for 1.5 hours. Then, 2 L of 40 V / V% methanol was passed for 1 hour to elute the adsorbed substance. The methanol solution was concentrated to obtain 25 g of a water-soluble soluble bayberry plant extract (referred to as product 1 of the present invention), which was a yellow solid. 1 mg was taken from the obtained solid matter and deionized water 1
It was dissolved by adding 0 ml and subjected to analysis of a water-soluble extract of the bayberry family. As a result, unlike the high-performance liquid chromatogram of the extract 1 as a raw material, the easily soluble soluble bayberry plant extract was composed of flavonol glycosides having many galactose residues transferred.

100 mg of the product 1 of the present invention was dissolved in 15 ml of 5% hydrochloric acid and heated at 100 ° C. for 3 hours for hydrolysis. After cooling to 20 ℃
The mixture was neutralized to pH 5 with a 5% sodium hydroxide solution, allowed to stand overnight in a refrigerator, and then separated into a precipitate and a filtrate. The precipitate was used for an aglycone analysis sample, and the filtrate was used for sugar analysis. The analysis of the precipitate and the sugar was performed using a high performance liquid chromatograph under the following conditions.

Analysis of precipitate Column: Microbonder pack C18, column diameter 4.6 m
m, column length 250 mm Solvent: Methanol / 5% acetic acid aqueous solution = 2/3 Flow rate: 1 ml / min Detector: UV / Visible spectrophotometer Measurement wavelength: 350 nm As a result of analysis, it is equivalent to a water soluble soluble bayberry plant extract. The peak disappeared, and flavonoid peaks such as myricetin were detected as aglycones. Further, the analysis of this filtrate was carried out under the same conditions, but the peak corresponding to the water-soluble extract of the bayberry family disappeared.

Analysis of Filtrate Column: Chemically modified aminopropyl silica (manufactured by Tokyo Chemical Industry Co., Ltd .; Kaseisorb LC NH2 Super) Column diameter 4.
6 mm, column length 250 mm Solvent: Acetonitrile / water = 4/1 Flow rate: 1 ml / min Detector: Differential refractometer As a result of analysis, the peak corresponding to galactose is the main component from the filtrate, and it corresponds to rhamnose and a small amount of glucose. Three peaks were detected. The composition ratio of galactose, rhamnose and glucose was determined from the comparison of the peak areas with the separately prepared standard solution, and the ratio was 2.4: 1: 0.2.

As is clear from the results of the high performance liquid chromatography analysis, it is clear that the product 1 of the present invention is a bayberry family extract in which 1 mol or more of galactose residues are transferred to the bayberry family plant extract.

Example 2 10 g of Extract 2 was suspended in 100 ml of hot water, 50% sodium hydroxide solution was added little by little to dissolve Extract 2, and then acetic acid was added to adjust the pH to 7.0 and then lactose. The temperature was adjusted to 65 ° C by adding 250 g. 0.5 g of β-galactosidase (enzyme titer 20,000) derived from Bacillus circulans was added to this solution to homogenize it, and the mixture was stirred at 65 ° C for 8 hours. Obtained reaction solution 0.0
2 ml was diluted with 2 ml of pure water and used for the analysis of the easily soluble waterberry plant extract. As a result of the analysis under the same conditions as in Example 1, the reaction solution was composed of an extract of the bayberry family in which many galactose residues were transferred. Then styrene-
Porous polymer composed of divinylbenzene copolymer
Purification was carried out in the same manner as in Example 1 using a column packed with 700 ml to obtain 15 g of a pale yellow solid substance, a water soluble soluble bayberry plant extract (referred to as product 2 of the present invention).

100 mg of this solid was hydrolyzed in the same manner as in Example 1, and the precipitate and the filtrate were analyzed by high performance liquid chromatography. As a result, the precipitate consisted of flavonoids such as myricetin, and the peak corresponding to the glycosided extract of the bayberry family disappeared from the filtrate. As a result of sugar analysis in the filtrate, galactose, rhamnose and glucose were detected, and their composition ratio was 2.5: 1:
Was 0.2. From the results of high-performance liquid chromatography analysis, it is clear that the product of the present invention is an extract of a plant belonging to the family Pediculaceae in which 1 mol or more of galactose residues are transferred.

Example 3 5.0 g of lactose was added to 5 ml of 0.05 M phosphate buffer (pH 6.8),
To this solution, 0.25 ml of dimethyl sulfoxide solution containing 50 mg of Extract 3 and 5 mg of β-galactosidase (enzyme titer of 20,000) derived from Bacillus circulans were added, and the temperature was 60 ° C.
And reacted for 5 hours. 0.02 ml of the obtained reaction solution was added to pure water 2
It was diluted with ml and subjected to analysis of transglycosylation reaction by high performance liquid chromatography under the same conditions as in Example 1. As a result, as in Example 1, a bayberry family plant extract having a large number of galactose residues transferred was detected. A water soluble soluble bayberry plant consisting of a pale yellow-brown powder was prepared by fractionating reaction products and unreacted substances under the following conditions using a preparative column packed with 100 liters of octadecylsilylated silica gel diluted with 1 liter of water. 15 mg of the extract (referred to as product 3 of the present invention) was obtained.

Solvent: 15% tetrahydrofuran aqueous solution Flow rate: 50 ml / min Detector: UV / visible spectrophotometer Measurement wavelength: 350 nm 5 mg of reaction product was hydrolyzed in the same manner as in Example 1 to obtain a precipitate and a solution. They were separated and subjected to high performance liquid chromatography analysis. As a result, flavonoids such as myricetin, which is an aglycone of flavonoid glycoside, were detected in the precipitate. As a result of sugar analysis, galactose, rhamnose and glucose were detected, and their composition was 2.6: 1: 0.2.
Met. These facts prove that the product of the present invention is a transfer of galactose residues to an extract of a plant belonging to the family Asaceae.

Example 4 To 1 ml of 0.05 M phosphate buffer (pH 6.8) was added 1.5 g of soybean oligosaccharide, and to this solution was added 0.05 ml of dimethyl sulfoxide solution containing 25 mg of extract 2 and Bacillus. Circulance-derived β-galactosidase (enzyme titer 20,000) (1 mg) was added, and the mixture was reacted at 60 ° C. for 5 hours. Dilute 0.02 ml of the obtained reaction solution with 2 ml of pure water,
The galactose residue transfer was used for the analysis of the bayberry plant extract. As a result, the reaction product was composed of a bayberry plant extract having four galactose residues transferred.

In order to clarify the effect of the present invention, a comparison of solubility in pure water and a test of antioxidant effect were carried out. Test Example 1 Comparison of Solubility in Pure Water An aqueous solution obtained by heating and dissolving an aqueous solution of the product 1 of the present invention according to Example 1 and the product 2 of the present invention according to Example 2 and the extract 1 as a raw material in pure water at 4 ° C. The presence and amount of precipitation was observed when left for 1 week. Table 1 shows the results.

[0036]

[Table 1]

Test Example 2 Comparison of Antioxidant Effect Water-soluble soluble bayberry plant extracts prepared in Example 1, Example 2 and Example 3 and BH widely used in general.
T and natural vitamin E were prepared for comparative experiments. still,
As a result of high performance liquid chromatography analysis, natural vitamin E was composed of a mixture of α, β, γ and δ type isomers, and the total tocopherol content was 70%.

Using palm oil, the present invention product 1, the invention product 2, the invention product 3 and other antioxidants were added, and a comparative test of the antioxidant effect was conducted. This method used a Rancimat (automatic oil and fat stability tester) manufactured by Metrohm Co., Ltd. According to this principle, air is blown into heated oil and fat, and this air is then blown into pure water. Volatile secondary products are produced along with the oxidation of fats and oils. The air blown into the oil layer carries the secondary product and transfers it to the water layer. The conductivity of water changes accordingly. The conductivity is plotted against time, the inflection point of the obtained curve is determined, and this time is used as the induction time. The stability of fats and oils is determined by comparing the length of the induction time with the increase of the induction time as the stability of the fats and oils increases. As the measurement conditions, heating temperature 100 ℃, air flow 20L / hour, sample amount 3.5g
I went in. Inventive products 1, inventive products 2 and 3 which are antioxidants to be tested were dissolved in hydrous ethanol, and BHT was dissolved in ethanol and added to fats and oils. The results are shown in Table 1.

[0039]

[Table 2]

The product of the present invention exerted a stronger effect of delaying the rancidity as compared with other antioxidants with a small amount added to palm oil.

Test Example 3 An experiment was conducted under the same conditions as in Test Example 1 except that the addition amount of the antioxidant was changed by using corn oil. The results are shown in Table 2.

[0042]

[Table 3]

The product of the present invention exerted a stronger effect of delaying the rancidity induction time on corn oil as compared with other antioxidants.

Test Example 4 Using pure lard, the heating temperature was 110 ° C. as a measurement condition.
An experiment was conducted under the same conditions as in Test Example 2 except that The results are shown in Table 4.

[0045]

[Table 4]

The product of the present invention exerted a stronger effect of delaying the rancidity induction time on pure lard as compared with other antioxidants. From the above results, the modified bayberry plant extract of the present invention was found to contain vitamin E and the synthetic antioxidant BH.
It was found that it has a better antioxidant power than T.

Test Example 5 A horse mackerel was abdominal opened, the internal organs were removed, and the horse mackerel was washed with water and then immersed in fresh water to remove blood. The present invention product 1 added 0.01% 10
% Saline solution was prepared, and the blood-tipped horse mackerel was immersed in this saline solution for 1 hour and then dried with cold air to prepare dried horse mackerel. As a control product, the additive-free product of the present invention was prepared in the same manner. Each dried horse mackerel was wrapped in a polyethylene film and stored at 3 ° C for 1 month. After sensory evaluation after baking so that both sides have an average baking color, the product to which the product of the present invention was added did not have an oxidative odor of oil, but the product without the additive had an oxidative odor of oil and was deteriorated. Was. Immediately after preparation and after 2 weeks of storage, dried horse mackerel was finely chopped and extracted with ethyl ether using a Soxhlet extractor to obtain crude fat. The amount of peroxide in the obtained crude fat was measured by RB Koch, B. Stern and CG Ferrar.
i proposed the Rodan iron method (Archives of Biochemistry)
and Biophysics. 78, 165-179 (1958)). The results are shown in Table 5.

[0048]

[Table 5]

It is said that the oxidation starts when the absorbance becomes 0.3 or more. As is clear from these results, the dried horse mackerel soaked in the saline solution containing the product 1 of the present invention clearly suppressed the oxidation as compared with the one dipped in the saline solution without the additive.

Test Example 6 Raw noodles were mixed with commercial salad oil containing 0.01% of the product 1 of the present invention at 170 ±
Fried noodles were prepared by frying at 5 ° C for about 1 minute. Similarly, fried noodles fried with additive-free commercial salad oil were prepared as a control product. The fried noodles were put in a polyethylene bag and stored in an incubator at 50 ° C. for 1 month, and then taken out for sensory comparison. As a result, the one to which the product 1 of the present invention was added was almost the same as that immediately after production, but , Those without additives had an oxidative smell of oil and the taste was changed.

Test Example 7 670 g of ground beef and pork, 30 g of beef tallow, 50 g of bread crumbs, 50 g of whole egg,
170g onion, 7g salt, 3g beef extract, white pepper powder
0.01 g of 1.5 g, nutmeg 0.5 g and the product 2 of the present invention were added and mixed sufficiently, and molded. As a control product, the product of the present invention 2 without addition was similarly prepared. After baking so that both sides have an average brown color, put it in a polyethylene bag and keep it at -20 ° C.
It was stored in the freezer for 6 months, taken out, thawed, and subjected to sensory evaluation to find the product of the present invention 2. The added ones did not change much after the production, but the unadded ones had a smell of oil and burns, and the taste was slightly changed.

Test Example 8 50 g of cream cheese and 29 g of powdered sugar were kneaded into a cream, and 5 g of white sucrose and 2 g of 50% citric acid aqueous solution were added and mixed until smoother. Into this, 0.1 g of a 10% ethanol solution of the product 1 of the present invention, 1 g of a flavoring agent and 0.2 g of a coloring agent were added, and then soft flour 100 was added.
A mixture of g, baking powder 2 g, xanthan gum 0.5 g and vegetable oil 40 g was added, mixed lightly, let stand for 1 hour in the refrigerator, molded, and baked at 170 ° C. for 10 minutes to prepare a cookie. As a control product, a product to which the product 1 of the present invention was not added was similarly prepared and baked. When each was put in a polyethylene bag and stored at 35 ° C for 3 months, those without the product 1 of the present invention had an offensive odor derived from peroxide, but the added cookies were the same as those immediately after baking. It was

Test Example 9 A raw material for a medicated vanishing cream having the following formulation was prepared.
Each of A and B was heated to 80 ° C. and dissolved. While stirring A, B was added to A and emulsified. The internal temperature of the mixture is 40
Stirring was continued until the temperature reached ℃, and then transferred to a jar.
Similarly, a control product to which the product 3 of the present invention was not added was prepared. When each of them was stored at 35 ° C for 12 months, the product to which the product of the present invention was added was not different from the product immediately after preparation,
The non-additive product as a control product had an offensive odor due to the oxidation of oil.

Formulation of A vanishing cream (weight ratio) Polyethylene glycol monostearate 2.8 Sorbitan monopalmitate 1.2 Dehydrated lanolin 1.0 Squalane 3.0 Stearic acid 8.5 Olive oil 2.0 Cetanol 4.0 Vaseline 4.0 Diphenhydramine 1.0 Liquid paraffin 3.0 Methylparaben 0.1 B Invention product 3 2.5 Propylene glycol 5.0 Boric acid 0.3 Purified water 61.6

Test Example 10 A cold cream raw material having the following formulation was prepared. A,
Each B was heated to 82 ° C. and uniformly dissolved. While well stirring A, B was added to A and emulsified. As a control product, a product to which the product 1 of the present invention was not added was prepared in the same manner. Each was transferred to a wide-mouthed bottle and stored at 35 ° C for 6 months. When the product of the present invention 1 was added, there was no difference from that immediately after preparation, but when the product of the present invention 1 was not added, an off-flavor was produced due to oxidation of fats and oils. Was there.

Cold cream formulation (weight ratio) A sorbitan monostearate 1.0 sorbitan monoisostearate 4.0 batyl monostearate 1.0 liquid paraffin 25.0 cetanol 4.0 beeswax 15.0 vaseline 5. 0 Methylparaben 0.1 B Borax 0.8 Inventive product 1 0.5 Purified water 42.1

Test Example 11 Northern seafood powder 600 g, fish liver powder 50 g, casein 100 g,
α-starch 150g, vitamin-mineral mixture 100
g, and corn oil 50 g into a mixture of 10 of the product 1 of the present invention.
% Ethanol solution (3 g) was added, mixed well, granulated and dried to prepare a feed for cultured fish. The present invention product 1 as a control product
A feed to which was not added was similarly prepared. Put each in a plastic bag and store at 35 ℃ for 2 months,
The product to which the product 1 of the present invention was added showed almost no change from the product immediately after preparation, but the additive-free product of the control product discolored and had a strong oxidative odor of fats and oils.

[0058]

EFFECTS OF THE INVENTION The readily water-soluble extract of the bayberry family of the present invention has the following features as compared with the extract of the bayberry family as a raw material. (1) Very good solubility in water. (2) Strong antioxidant effect. (3) Since the solubility in water is improved, there is no problem of precipitate formation even if used in high concentrations in foods, pharmaceuticals, quasi-drugs, and cosmetics containing water. be able to. These characteristics are due to the extremely high solubility in water as a result of the transfer of galactose residues to the extract of the bayberry family.As a water-soluble antioxidant, foods, pharmaceuticals, and quasi drugs are used. , Can be used for cosmetics or feed.

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Claims (2)

[Claims] 1. A galactose residue is transferred to a bayberry plant extract by allowing an enzyme having a galactose residue transfer activity to act in the coexistence of a bayberry plant extract having an antioxidant effect and a galactose residue transfer source. A water soluble soluble bayberry plant extract characterized by the above. 2. The easily water-soluble bayberry plant extract according to claim 1, wherein the bayberry family plant extract is an extract of a plant of the family Pebra family of the family Pebrinaceae with an organic solvent.