JP3321731B2 - α-Glycosyl naringin, its production method and use - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel α-glycosyl naringin, a method for producing the same, and uses thereof. More specifically, α-glycosyl naringin in which a D-glucose residue is α-bonded to naringin in an equimolar amount or more, And a method for producing α-glycosyl naringin, which comprises reacting a solution containing naringin and an α-glucosyl sugar compound with a glycosyltransferase to produce a novel α-glycosyl naringin, and collecting the same, and Beverages characterized by containing the α- glycosyl naringin obtained by this method, food and drink such as processed foods,
Prophylactic or therapeutic agents for susceptibility diseases, ie, anti-sensitivity disease agents,
The present invention relates to a use for various tissues such as cosmetics such as skin-beautifying agents and fair-skin agents.

[0002]

2. Description of the Related Art Naringin has a chemical structure represented by the following chemical formula 1, and has long been known as a vitamin P having physiological actions such as strengthening of capillaries, prevention of bleeding and blood pressure regulation, and as a bittering agent. It is used for food, medicine, cosmetics, etc.

[0003]

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[0004] Vitamin P participates in vivo in the physiological activity of vitamin C, for example, the hydroxylation reaction of proline and lysine necessary for the synthesis of collagen, which is a main component of biological connective tissue. Fe ++
It is known to be involved in oxidation-reduction reactions such as reduction of + to Fe ++, and also to enhance those reactions, such as being involved in the immunopotentiating action due to increased leukocytes. Plays an important role.

[0005] The use of naringin is not limited to vitamin P fortifier as a nutrient, but it may be used alone or in combination with other vitamins due to its chemical structure and physiological action. Agents, quality-improving agents, ultraviolet absorbers, etc., in foods and drinks, as well as in prophylactic and therapeutic agents for susceptible diseases such as viral diseases, bacterial diseases, cardiovascular diseases, malignant tumors, ie, anti-sensitive disease agents, and Has a wide range of cosmetics as a beautiful skin agent such as a stabilizer, an antioxidant, an ultraviolet absorber, a melanin production inhibitor, and a skin lightening agent.

[0006] However, naringin is poorly soluble in water,
At room temperature 25 ° C., only 1 g (about 0.1 w
/ V%), which is extremely difficult to use.

[0007]

SUMMARY OF THE INVENTION A naringin derivative which solves the drawbacks of conventional naringin, has excellent water solubility, maintains bitterness, has no concern about toxicity, and has a sufficient biological activity in vivo. Realization is strongly desired.

[0008]

DISCLOSURE OF THE INVENTION The present invention has been made to solve the above-mentioned drawbacks, and in particular, has made intensive studies for a novel naringin derivative by utilizing a biochemical technique.

As a result, the glycosyltransferase is allowed to act on a solution containing naringin and an α-glucosyl sugar compound, so that it is excellent in water solubility, maintains bitterness, has no toxicity, and is easily hydrolyzed in vivo. It has been found that it is possible to produce a novel α-glycosyl naringin that can be decomposed to exhibit the original physiological activity of naringin, and its production method and its use in various compositions such as foods and drinks, preventive agents for susceptible diseases, therapeutic agents, and cosmetics. Established and completed the present invention.

[0010] Further, α-
In purifying glycosyl naringin, it was found that the reaction solution was brought into contact with a porous synthetic adsorbent and the difference in the adsorptivity was utilized to easily purify the glycosyl naringin, and the present invention was completed.

Therefore, the method of the present invention for producing α-glycosyl naringin eliminates the drawbacks of the prior art at once, and makes it very easy to realize industrialization.

Hereinafter, the present invention will be described in detail.

The naringin used in the present invention need not be limited to highly purified naringin, but may be a mixture of naringin and flavonoid glycosides such as citronin, hesperidin, rutin, etc. Extracts derived from various plants or partially purified products thereof can be used as appropriate. As the plant tissue, for example, citrus fruits, pericarp, immature fruits and the like can be advantageously used.

The α-glucosyl sugar compound used in the present invention can be converted from naringin to α-glucosyl by the simultaneously used glycosyltransferase.
Any substance capable of producing glycosyl naringin may be used. For example, starch partially hydrolyzed products such as amylose, dextrin, cyclodextrin, and maltooligosaccharides, and further, liquefied starch and gelatinized starch are appropriately selected.

Accordingly, in order to facilitate the production of α-glycosyl naringin, an α-glucosyl sugar compound suitable for a glycosyltransferase is selected.

For example, when α-glucosidase (EC 3.2.1.20) is used as a glycosyltransferase, maltooligosaccharides such as maltose, maltotriose, and maltotetraose, or DE of about 10 to 70 A partial hydrolyzate of starch and the like are preferable, and cyclomaltodextrin glucanotransferase (EC 2.4.
When 1.19) is used, cyclodextrin or a starch hydrolyzate having a DE of about 60 from a starch gelatinized product having a DE of 1 or less is preferable, and α-amylase (EC3.
When 2.1.1) is used, starch gelatinized products having a DE of 1 or less, dextrin having a DE of about 30 and partially hydrolyzed starch are preferred.

The concentration of the α-glucosyl sugar compound at the time of the reaction is preferably about 0.5 to 100 times, preferably about 2 to 20 times that of naringin. The naringin-containing liquid at the time of the reaction is desirably one containing naringin as high as possible. For example, naringin is dissolved in suspension or at a high temperature, or at an alkaline pH exceeding pH 7.0. It is suitable to use a solution containing a high concentration in the form of a solution having a concentration of about 1.0 w / v%.
The above high concentration, desirably about 2.0 to 20.0 w /
means a solution containing v%. At this time, naringin in the alkaline solution is liable to be decomposed. Therefore, in order to prevent this, it is desirable that the naringin be kept as light-shielded and as anaerobic as possible. If necessary, an antioxidant such as L-ascorbic acid and erythorbic acid may be co-present.

The glycosyltransferase used in the present invention is one which produces α-glycosyl naringin without decomposing naringin when acted on a solution containing naringin and an α-glucosyl sugar compound having properties suitable for this enzyme. Should be fine. For example, α-glucosidase is an enzyme derived from animal or plant tissues such as pig liver, buckwheat seed, or a genus Mucor, Penicillium.
um) or a fungus belonging to the genus Saccharomyces (S
An enzyme derived from a culture obtained by culturing a microorganism such as yeast belonging to the genus Accharomyces or the like in a nutrient medium is cyclomaltodextrin glucanotransferase. Α-amylase is a bacterium belonging to the genus Bacillus, or Aspergillus (Aspergi).
For example, an enzyme derived from a mold culture belonging to the genus lux or the like can be appropriately selected.

These glycosyltransferases do not necessarily need to be purified and used as long as the above conditions are satisfied.
Usually, the purpose of the present invention can be achieved with a crude enzyme. If necessary, it may be used after being purified by various known methods. Alternatively, a commercially available glycosyltransferase can be used. There is a close relationship between the amount of enzyme used and the reaction time, and usually the amount of enzyme is selected so that the reaction is completed in about 5 to 80 hours from the viewpoint of economy.

The immobilized glycosyltransferase may be used in a batch-wise or continuous manner for the reaction.

Further, in order to prevent the decomposition of naringin in the reaction solution, it is desirable to carry out the reaction under light shielding, anaerobic conditions and slightly acidic conditions as much as possible.

The relatively high-molecular-weight α-glycosyl naringin produced by the transglycosylation reaction may be purified as it is or after purification with a porous synthetic adsorption resin, if necessary, with glucoamylase (EC 3.2.1). .3)
Alternatively, partial hydrolysis by an amylase such as β-amylase (EC 3.2.1.2) can reduce the number of α-D-glucosyl groups of α-glycosylnarindin. For example, when glucoamylase is allowed to act, it is possible to hydrolyze a high molecular substance equal to or more than α-maltosyl naringin to generate glucose and accumulate and generate α-glucosyl naringin.
When amylase is allowed to act, it is possible to hydrolyze high-molecular substances of α-maltotriosyl naringin or more to generate maltose and to accumulate and generate a mixture of mainly α-glucosyl naringin and α-maltosyl naringin. .

The reaction solution that has produced α-glycosyl naringin as described above can be directly used as an α-glycosyl naringin product. Usually, the reaction solution is filtered, concentrated and syrup-shaped, and further dried,
Powdered into powdered α-glycosyl naringin product.

This product is used not only as a vitamin P enhancer but also as a highly safe natural bittering agent, antioxidant, stabilizer, quality improving agent, preventive agent, therapeutic agent, ultraviolet absorber, etc. It can be advantageously used for various compositions such as foods, foods, feeds, feeds, anti-sensitivity agents, cosmetics, and plastic products.

Further, when producing a purified α-glycosyl naringin product, α-glycosyl naringin and α-glycosyl naringin are utilized by utilizing the difference in adsorptivity of the porous synthetic adsorbent.
-It may be separated and purified from impurities such as glucosyl sugar compounds.

The porous synthetic resin referred to in the present invention is a porous synthetic resin having a large adsorption surface area and a non-ionic styrene resin.
It is a synthetic resin such as a divinylbenzene polymer, a phenol-formalin resin, an acrylate resin, and a methacrylate resin.
Amberlite XAD-1, Amberlite XAD-2, Amberlite XAD-4 manufactured by Haas
Amberlite XAD-7, Amberlite XAD-
8, Amberlite XAD-11, Amberlite XA
D-12, trade name manufactured by Mitsubishi Kasei Kogyo Co., Ltd. Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, trade name IMACTI manufactured by IMACTI S
yn-42, Imakti Syn-44, Imakti Sy
n-46.

The method of purifying a reaction solution that has produced α-glycosyl naringin according to the present invention is characterized in that, when the reaction solution is passed through, for example, a column filled with a porous synthetic adsorbent, α-glycosyl naringin and a relatively small amount of While unreacted naringin is adsorbed on the porous synthetic adsorbent, a large amount of coexisting α-glucosyl sugar compound and water-soluble saccharide flow out without being adsorbed. If necessary, after the completion of the glycosyltransferase reaction and before contact with the porous synthetic adsorbent, for example, the reaction solution is heated to remove insolubles generated by filtration, or magnesium aluminate silicate, alumina It is treated with magnesium acid or the like to adsorb and remove proteinaceous substances and the like in the reaction solution, and ion exchange resins (H type and OH
It is optional to use a combination of purification methods such as desalting after treatment with a compound (type).

As described above, the α-glycosyl naringin selectively adsorbed on the porous synthetic adsorbent column and a relatively small amount of unreacted naringin are washed with a dilute alkali, water or the like, and then washed with a relatively small amount. When an organic solvent or a mixture of an organic solvent and water, for example, methanol water or ethanol water, is passed, first, α-glycosyl naringin is eluted, and it is not possible to increase the flow rate or increase the concentration of the organic solvent. Reaction naringin elutes. The eluate having a high content of α-glycosyl naringin is subjected to a distillation treatment, an organic solvent is first distilled off, and then concentrated to an appropriate concentration to obtain a syrup-like product containing α-glycosyl naringin as a main component. Furthermore, by drying and powdering this,
A powdered product based on α-glycosyl naringin is obtained. The elution operation of α-glycosyl naringin and unreacted naringin with the organic solvent is performed simultaneously,
Since the operation becomes a regeneration operation of the porous synthetic adsorbent, the porous synthetic adsorbent can be repeatedly used.

Further, the purification using the porous synthetic adsorbent of the present invention has a feature that not only α-glucosyl sugar compounds and water-soluble saccharides but also impurities such as water-soluble salts can be removed at the same time. The α-glycosyl naringin thus obtained has the following features.

(1) Compared with naringin, α-glycosyl naringin has extremely high water solubility. (2) Compared with naringin, α-glycosyl naringin has higher light fastness and stability. (3) α-glycosyl naringin has substantially the same bitterness as naringin. (4) α-Glycosyl naringin is hydrolyzed by an enzyme in the body into naringin and glucose, and exhibits the original physiological activity (vitamin P) of naringin. Also, α-lycosyl naringin is used as an antioxidant for other vitamins such as thiamine, riboflavin, vitamin C, and vitamin E.
It is also convenient to use them in combination with one or more species. In addition, the combined use with vitamin C can enhance their physiological activity. (5) In the case of a product containing an α-glucosyl sugar compound, not only the α-glycosyl naringin can exert the effect, but also the α-glucosyl sugar compound can exert the shaping, bulking and sweetening effects. In the case of a purified product from which the α-glucosyl sugar compound has been removed,
The effect of α-glycosyl naringin can be exhibited without increasing the amount.

Due to these features, α-glycosyl naringin is not only a highly safe natural vitamin P enhancer, but also a bittering agent, an antioxidant, a stabilizer, a quality improving agent, a viral disease, and a bacterial disease. As foods, foods, foods, feeds, feeds, anti-sensitivity agents, skin-whitening agents, skin-whitening agents, cosmetics such as circulatory diseases, malignant tumors, etc. It can be advantageously used for various compositions such as products.

The taste of α-glycosyl naringin is in harmony with various substances exhibiting sourness, astringency, umami, etc. from acidity and salt, and has high acid resistance and heat resistance. Goods, for example, seasonings, Japanese sweets, western confectionery, frozen desserts, beverages, spreads, pastes, pickles, canned bottles, processed meat, processed fish and marine products, processed milk and eggs, processed vegetables, processed fruits, processed cereals It can be widely used for goods. In particular, it is also convenient to impart the bitterness exhibited by α-glycosyl naringin to citrus fruits, cocoa, coffee, chocolate, teas, herbal medicines, or flavors thereof to improve the flavor of various foods and drinks. In addition, α-glycosyl naringin is a plant-derived sweetener such as stevioside, α-glycosyl stevioside, rebaudioside A, glycyrrhizin, α-glycosyl glycyrrhizin, dihydrochalcones, amino acid sweeteners such as glycine, alanine, L-asparatyl phenylalanine methyl ester, Sucrose, syrup, sugar-bound syrup, glucose, isomerized sugar, fructose, honey, maltose, sorbitol, maltitol,
It can also be advantageously used in combination with one or two or more sweeteners selected from sugar sweeteners such as lactose for the purpose of improving taste and flavor. Further, since α-glycosyl naringin inhibits crystallization and precipitation of the flavonoid compound in the juice, it can be advantageously used as an antifoggant, an antifogging agent, and the like for fruit juice drinks, fruit juice jelly and the like. In addition, livestock, poultry, bees, silkworms,
Vitamin P in feed and feed for breeding animals such as fish
It is also convenient to mix and use them for the purpose of enhancing agents and improving palatability.

In addition, tobacco, troches, liver oil drops, multivitamin preparations, oral fresheners, oral incense tablets, gargles, tube feedings, crude drugs, oral medications, injections, toothpaste,
Lipstick, lip balm, sunscreen, etc. various solid, paste, liquid taste, preventive and therapeutic agents for susceptible diseases,
In other words, it can be advantageously used by blending it into cosmetics such as anti-sensitivity disease agents, skin-beautifying agents, and skin-whitening agents, and further, it is blended and used in plastic products and the like as an ultraviolet absorber, a deterioration inhibitor and the like. This can also be carried out advantageously.

The susceptibility disease referred to in the present invention is α-
A disease that is prevented or treated by glycosyl naringin, such as a viral disease,
It may be a bacterial disease, traumatic disease, immune disease, rheumatism, diabetes, circulatory disease, malignant tumor and the like. α-glycosyl naringin susceptible disease preventive agent, therapeutic agent,
The shape can be freely selected according to the purpose. For example, sprays, eye drops, nasal drops, gargles, solutions such as injections, ointments, cataplasms, pastes such as creams,
Solid preparations such as powders, granules, capsules and tablets.
In the formulation, if necessary, other components, for example,
It is optional to use one or more of a therapeutic agent, a physiologically active substance, an antibiotic, an auxiliary agent, a bulking agent, a stabilizer, a coloring agent, a flavoring agent and the like. The dose can be appropriately adjusted depending on the content, administration route, administration frequency and the like. Usually α
Glycosyl naringin in the range of about 0.001 to 10.0 grams per adult daily is preferred. Also, in the case of cosmetics, it can be used according to the above-mentioned prophylactic and therapeutic agents.

As a method of utilizing α-glycosyl naringin, known processes such as kneading, kneading, dissolving, dipping, penetrating, spraying, coating, spraying, and pouring are used in the steps until the products of the various compositions are completed. The method is appropriately selected.

Hereinafter, an example of the α-glycosyl naringin of the present invention will be described by experiments.

(Experiment 1) <Preparation of α-glycosyl naringin> (1) Glycosyl transfer reaction 1 part by weight of naringin and dextrin (DE20) 6
50 parts by weight of water was added to the parts by weight and melted by heating, and the mixture was dissolved in Bacillus stearothermophilus (Bacillus st.
Cyclomaltodextrin glucanotransferase (produced by Hayashibara Biochemical Laboratories, Inc.) derived from E. thermothermophilus was added at a rate of 2 per dextrin rum.
0 units were added, the reaction was carried out for 18 hours while maintaining the pH at 6.0 and 70 ° C., and then the mixture was inactivated by heating to obtain an α-glycosyl naringin-containing liquid.

(2) Purification The reaction solution obtained by the method of (1) was filtered, and the filtrate was applied to a column filled with a porous synthetic adsorbent, Diaion HP-10 (trade name, sold by Mitsubishi Chemical Corporation). The liquid was passed.
After washing the column with water, 50 v / v% ethanol was passed therethrough, the eluate was concentrated, the solvent was distilled off, the powder was pulverized, and pale yellow α-glycosyl naringin standard [I] was used as the starting naringin. Obtained in a yield of about 130% by weight.

(3) Hydrolysis by Amylase The α-glycosyl naringin standard [I] obtained by the method of (2) is dissolved in water at 1 w / v%, and glucoamylase (EC 3.2.1. 3, sold by Seikagaku Corporation), 100 units per gram of the sample, pH 5.0,
The reaction was maintained for 5 hours at 55 ° C. The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was passed through a column of a porous synthetic adsorbent, Diaion HP-10 (trade name, sold by Mitsubishi Chemical Industry Co., Ltd.) at SV2. As a result, the α-glycosyl naringin and the unreacted naringin in the solution were adsorbed by the porous synthetic adsorbent, and glucose, salts, etc. were flown out without being adsorbed. Next, the column was passed through with water and washed, and then passed while increasing the concentration of the aqueous ethanol solution in a stepwise manner. The α-glycosyl naringin fraction was collected, concentrated under reduced pressure, and powdered to obtain pale yellow α-glycosyl. The naringin standard [II] was obtained in a yield of about 70% based on the weight of the naringin as the raw material per solid.

Also, instead of glucoamylase, β-amylase (EC 3.2.1.2) and (available from Seikagaku Corporation) were used to prepare α-glycosylnarindin standard according to the method described above. [I] was hydrolyzed, purified, concentrated and powdered to obtain a pale yellow α-glycosyl naringin standard [III] in a yield of about 70% based on the weight of the raw material naringin.

(Experiment 2) <Physicochemical properties of α-glycosyl naringin> (1) Improvement of water solubility α-glycosyl naringin-containing solution prepared by performing a transglycosylation reaction according to the method of Experiment 1 (1) and used in this method The enzyme was heated and inactivated in advance and treated according to the method of Experiment 1 (1). The control solution was allowed to stand in a cold room at 4 ° C. for 2 days. In contrast, the solution containing α-glycosyl naringin remained transparent.

Accordingly, α-glycosyl naringin produced by the transglycosylation reaction is highly water-soluble and is improved.

(2) Solubility in Solvent The α-glycosyl naringin standard is easily soluble in water, 0.1N sodium hydroxide, 0.1N hydrochloric acid, slightly soluble in methanol and ethanol, and insoluble in ether, benzene and chloroform.

(3) Taste Taste naringin, α-glycosyl naringin [I],
Each of the sample [II] and the sample III] was used as a 1 mM aqueous solution, and the taste was compared. The samples [I] and [I]
It was found that both I] and the preparation [III] maintain the bitterness of the raw material naringin well and show almost the same bitterness as that of the raw material naringin.

(4) Ultraviolet absorption spectrum An aqueous solution of α-glycosyl naringin sample (pH 6.
When the ultraviolet absorption spectrum was examined using 0), all of the sample [I], the sample [II] and the sample [III] showed 211 n in the order of the absorption intensity similarly to the case of naringin.
m, absorption maxima at around 282 nm and 329 nm.

(5) Infrared absorption spectrum The infrared absorption spectrum of the α-glycosyl naringin sample was examined by the KBr tablet method. FIG. 1 shows the results of the α-glycosyl naringin standard [II]. FIG. 2 shows the results of nalinn as a control.

(6) Stability to Hydrolysis (a) α-Glycosyl naringin preparation is α-glucosidase derived from pig liver (EC 3.2.1.20)
To produce naringin and D-glucose.

(B) Not hydrolyzed by β-glucosidase.

[0049]

(B) Results of analysis When the α-glycosyl naringin sample was analyzed, in the case of the sample [I], in addition to the naringin spot of Rf 0.79, Rf 0.68, 0.49, 0.3
A few spots to the origin are recognized along with 8, 0.21, etc., and in the case of the sample [II], spots of Rf 0.68 are recognized, and in the case of the sample [III], Rf 0.6
8, 0.49 spots were observed.

From the above physicochemical properties, the substance exhibiting Rf 0.68 contained in the sample [I], the sample [II] and the sample [III] has one mole of naringin and one mole of D-glucose residue. It was determined to be α-linked α-glucosyl naringin, and Rf0.
The substance showing 49 was determined to be α-maltosyl naringin in which 2 moles of D-glucose was α-bonded to 1 mole of naringin. Similarly, the substance showing a plurality of spots with Rf 0.38 or less contained in the sample [I] was , 1 mole of naringin D
-It is judged to be α-maltooligosyl naringin in which glucose residues are α-bonded by 3 mol or more.

Thus, the α-glycosyl of the present invention
Naringin is a novel naringin sugar derivative having good water solubility in which D-glue residue is α-bonded to naringin in equimolar or more. When ingested into a living body, it is easily hydrolyzed by α-glucosidase, and the natural physiological properties of naringin Demonstrate activity.

(Experiment 3) <Acute toxicity> Experiment 1 was performed using a 7-year-old dd mouse.
An acute toxicity test was conducted by oral administration of the α-glycosyl naringin preparation [I] prepared by the method of (2).
No deaths were observed up to 5 g per kg of body weight. Therefore, the toxicity of this substance is extremely low.

It should be noted that α-prepared by the method of Experiment 1 (3)
When this test was performed using glycosyl naringin preparation [II], similar results were obtained, and it was found that the toxicity was extremely low.

Hereinafter, as examples of the present invention, a production example of α-glycosyl naringin will be described in Example A, and application examples of α-glycosyl naringin in various compositions will be described in Example B.

(Example A-1) <α-Glycosyl naringin> 1 part by weight of naringin and 4 parts by weight of dextrin (DE10) were dissolved in 10 parts by weight of water while heating, and immediately cooled to 75 ° C., followed by Bacillus stearothermophilus. Cyclomaltodextrin glucanotransferase (available from Hayashibara Biochemical Laboratory Co., Ltd.) was added at 20 units per dextrin trauma, pH
The reaction was maintained at 5.5 and 75 ° C. for 24 hours with stirring. When the reaction solution was analyzed by thin-layer chromatography, about 65% of the naringin was converted to α-glycosyl naringin such as α-glucosyl naringin, α-maltosyl naringin, α-maltotriosyl naringin, α-maltotetraosyl naringin. Had been converted.
The reaction solution is heated to inactivate the enzyme, filtered, and the filtrate is desalted and purified using an ion exchange resin (H type and OH type) according to a conventional method, and concentrated to obtain a syrup-like α-glucosyl sugar compound. The contained α-glycosyl naringin product was obtained in a yield of about 85% by weight of the raw material per solid.

This product is used not only as a vitamin P enhancer but also as a highly safe natural bittering agent, antioxidant, stabilizer, quality improving agent, preventive agent, therapeutic agent, ultraviolet absorber, etc. Food, food, feed, feed, anti-sensitivity drug,
It can be advantageously used for various compositions such as cosmetics and plastic products.

(Example A-2) <α-Glucosyl naringin> One part by weight of an α-glycosyl naringin product having a syrup-like α-glucosyl sugar compound prepared according to the method of Example A-1 was mixed with 4 parts of water.
Glucoamylase (EC 3.
2.1.3, sold by Seikagaku Corporation), 100 units per gram of α-glycosyl naringin product solids,
The reaction was performed at 50 ° C. for 5 hours. When the reaction solution was analyzed by thin layer chromatography, α-glycosyl naringin was converted to α-glucosyl naringin. The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was passed through a column of a porous synthetic adsorbent, Diaion HP-10 (trade name, sold by Mitsubishi Chemical Industry Co., Ltd.) at SV2. As a result, α-glucosyl naringin and unreacted naringin in the solution were adsorbed on the porous synthetic adsorbent, and glucose, salts, etc., flowed out without being adsorbed. Then, the column was passed through and washed with water, and then passed while increasing the concentration of the aqueous ethanol solution in a stepwise manner. The α-glucosyl naringin fraction was collected, concentrated under reduced pressure, powdered, and powdered α-glucosyl Naringin was obtained in a yield of about 55% based on the weight of the raw material naringin per solid. When α-glucosyl naringin was hydrolyzed with an acid, 1 mole of L-rhamnose and 2 moles of D-glucose were produced for 1 mole of naringenin, and α-glucosyl naringin was extracted from pig liver and partially purified. It was found that when α-glucosidase was acted on, it was hydrolyzed to naringin and D-glucose.

The α-glucosyl naringin can be used as a highly purified water-soluble vitamin P enhancer, as well as a bittering agent, an antioxidant, a stabilizer, a quality improving agent, a prophylactic agent, a therapeutic agent, and an ultraviolet absorber. For example, it can be advantageously used for various compositions such as foods and drinks, favorite foods, anti-sensitivity agents, cosmetics and the like.

(Example A-3) <α-Glycosyl naringin> 1 part by weight of naringin and 10 parts by weight of dextrin (DE8) were dissolved by heating in 20 parts by weight of water, cooled to 70 ° C., and added with cyclomaltodextrin gluca. No transferase was added in an amount of 30 units per dextrin, and the mixture was reacted at pH 6.0, maintained at 65 ° C., and stirred for 40 hours. When the reaction solution was analyzed by thin-layer chromatography, it was found that
Was converted to α-glycosyl naringin. The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was filtered using a porous synthetic adsorbent, trade name Amberlite XAD-7 (Rohm &
(Manufactured by Haas) at SV 1.5.

As a result, α-glycosyl naringin and unreacted naringin in the solution were adsorbed on the porous synthetic adsorbent, and dextrin, oligosaccharides, salts, etc. were flowed out without being adsorbed. The column was washed with water and washed with water.
0-v / v% methanol was passed through to elute α-glycosyl naringin and unreacted naringin, which were concentrated and pulverized to obtain a powdered α-glycosyl naringin product of about 110% based on the weight of the raw material naringin. In a yield of

The product is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural bittering agent, antioxidant, stabilizer, quality improving agent, prophylactic agent, therapeutic agent, ultraviolet absorber. For example, it can be advantageously used for various compositions such as foods and drinks, foods, feeds, feeds, anti-sensitivity agents, cosmetics, and plastic products.

Example A-4 <α-Glycosyl naringin> (1) Preparation of α-glucosidase preparation Maltose 4% w / v, monopotassium phosphate 0.1% w / v
%, Ammonium nitrate 0.1 w / v%, magnesium sulfate 0.05 w / v%, potassium chloride 0.05 w / v%, polypeptone 0.2 w / v%, calcium carbonate 1 w / v%
(Separately sterilized by dry heat and aseptically added at the time of inoculation) and 500 parts by weight of a liquid medium consisting of water and Mucor javanicus IFO 4570
Was shake-cultured at a temperature of 30 ° C. for 44 hours. After completion of the culture, the mycelium was collected, and 0.5 parts with respect to 48 parts by weight of the wet mycelium.
4M urea solution 50 dissolved in M acetate buffer (pH 5.3)
After adding 0 parts by weight, the mixture was allowed to stand at 30 ° C. for 40 hours, and then centrifuged. The supernatant was dialyzed overnight in running water, and then ammonium sulfate was added.
The mixture was left to stand at 4 ° C. overnight at 9 ° C., and the formed salt precipitate was collected by filtration, suspended and dissolved in 50 parts by weight of 0.01 M acetate buffer (pH 5.3), and then centrifuged to collect the supernatant. Then, an α-glucosidase preparation was obtained.

(2) Preparation of α-glycosyl naringin 3 parts by weight of naringin and dextrin (DE30) 2
0 parts by weight are heated and dissolved in 50 parts by weight of water, cooled to 55 ° C., and immediately after cooling to 55 ° C., the α-glucosidase preparation 1 prepared by the method of (1) is used.
0 parts by weight, maintaining the pH at 6.0 and stirring with 55 parts by weight.
Reaction was performed at 40 ° C. for 40 hours. The reaction solution was analyzed by thin-layer paper chromatography.
Was converted to α-glycosyl naringin. The reaction solution was purified, concentrated and powdered in the same manner as in Example A-3 to obtain a powdery α-glycosyl naringin product in a yield of about 95% based on the weight of the raw material naringin.

This product is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural-type bittering agent, antioxidant, stabilizer, quality improvement, as in Example A-3. It can be used for various compositions as an agent, a prophylactic agent, a therapeutic agent, an ultraviolet absorber and the like.

(Example B-1) <Hard candy> 1,500 parts by weight of reduced maltose starch syrup (manufactured by Hayashibara Shoji Co., Ltd., registered trademark: Mavit) are heated and concentrated under reduced pressure until the water content becomes about 2% or less. Then, 1 part by weight of the powdery α-glycosyl naringin obtained by the method of Example A-3 and an appropriate amount of caramel and coffee flavor were mixed, followed by molding and packaging according to a conventional method to obtain a hard candy.

This product is a coffee candy with enhanced vitamin P and bitterness, and has low caries and low calories.

(Example B-2) <Boiled Butterfly> Butterfly was peeled, cut into appropriate lengths, immersed in a thin saline solution for several hours, and then subjected to α-glycosyl.
Boiled in a liquid containing a green colorant prepared by mixing rutin and Blue No. 1 and a syrup-like α-glycosyl naringin obtained by the method of Example A-1, and boiled green fresh butterbur in water. Obtained.

This product has a bitter taste rich in wildness,
It is also suitable as a material for various Japanese dishes.

(Example B-3) <Fertilization> 1 part by weight of waxy seed starch was mixed with 1.2 parts by weight of water, and gelatinized by heating, and 1.5 parts by weight of sugar and crystalline β
-0.7 parts by weight of maltose (manufactured by Hayashibara Co., Ltd., sun malt), 0.3 parts by weight of starch syrup, and Example A-
Then, 0.02 parts by weight of the syrup-like α-glycosyl naringin obtained by the method of Example 1 was mixed, and then molded and formed according to a conventional method.
Packed to produce fertilizer.

This product is a kibidan-style Japanese confectionery with a rich and subtle bitter taste and a good mouthfeel.

(Example B-4) <Mixed sweetener> 100 parts by weight of honey, 50 parts by weight of isomerized sugar, 1 part by weight of α-glycosyl stevioside (manufactured by Toyo Sugar Co., Ltd., trade name α-G suite) and 0.02 parts by weight of the powdery α-glycosyl naringin obtained by the method of Example A-4 was mixed to obtain a mixed sweetener.

This product is a sweetener with good taste and enhanced vitamin P. Its sweetness is about twice that of sugar and is suitable as a health food.

(Example B-5) <Sand cream> 1,200 parts by weight of crystalline α-maltose (manufactured by Hayashibara Co., Ltd., fine tose)
1,000 parts by weight of shortening, 50 parts by weight of cacao mass, powdery α-glycosyl obtained by the method of Example A-3
3 parts by weight of naringin and 1 part by weight of lecithin were mixed by a conventional method to produce a sand cream.

This product is a chocolate-flavored sand cream with enhanced vitamin P and increased bitterness.

(Example B-6) <Tablet> 20 parts by weight of L-ascorbic acid, 13 parts by weight of crystalline β-maltose, 4 parts by weight of cornstar and powdered α-form obtained by the method of Example A-2 After uniformly mixing 3 parts by weight of glucosyl naringin, diameter 12 mm, 20R
Tablets were obtained using a pestle to obtain tablets.

This product is a multivitamin of L-ascorbic acid and α-glucosyl naringin, and is a tablet with good stability of L-ascorbic acid and easy to swallow.

(Example B-7) <Capsule> 10 parts by weight of calcium acetate monohydrate, L
-50 parts by weight of magnesium lactate / trihydrate, matose 57
Parts by weight, 20 parts by weight of α-glucosyl naringin obtained by the method of Example A-2 and 12 parts by weight of a γ-cyclodextrin clathrate compound containing 20% of eicosapentaenoic acid were uniformly mixed and granulated by a granulator. Then, according to a conventional method, the capsule is encapsulated in a gelatin capsule, and one capsule 150
mg capsules were prepared.

The product can be advantageously used as a blood cholesterol lowering agent, an immunopotentiator, a skin-beautifying agent, a preventive agent for susceptibility diseases, a therapeutic agent, a food for promoting health, and the like.

(Example B-8) <Ointment> 1 part by weight of sodium acetate / trihydrate and 4 parts by weight of DL-calcium lactate were uniformly mixed with 10 parts by weight of glycerin, and this mixture was mixed with 50 parts by weight of petrolatum, Tree wax 10
Parts by weight, lanolin 10 parts by weight, sesame oil 14.5 parts by weight,
In addition to a mixture of 1 part by weight of α-glycosyl naringin obtained by the method of Example A-4 and 0.5 part by weight of peppermint oil,
The mixture was further uniformly mixed to produce an ointment.

The product can be advantageously used as a sunscreen, a skin-beautifying agent, a skin-whitening agent, etc., and also as an accelerator for healing wounds and burns.

Example B-9 <Injection> α-Glucosyl obtained by the method of Example A-2
Naringin was dissolved in water and purified and filtered to be pyrogen-free according to a conventional method. This solution was dispensed into a 20-mL ampoule so that the amount of α-glucosyl naringin became 50 mg, and this was dried under reduced pressure, sealed, and injected. Was manufactured.

This injection can be administered intramuscularly or intravenously either alone or as a mixture with other vitamins and minerals. In addition, the product does not need to be stored at low temperature, and has extremely good solubility in physiological saline and the like when used.

(Example B-10) <Injection> 6 parts by weight of sodium chloride, 0.1 part of potassium chloride.
3 parts by weight, 0.2 parts by weight of calcium chloride, 3.1 parts by weight of sodium lactate, 45 parts by weight of maltose and Example A-
Dissolve 1 part by weight of α-glucosyl naringin obtained in method 2 in 1,000 parts by weight of water, purify and filter it according to a conventional method to make it pyrogen-free, fill the sterilized plastic container with 250 mL each and inject it. An agent was manufactured.

This product is an injection for supplementing calories and minerals as well as supplementing vitamin P, and can be advantageously used for promoting treatment during and after illness and promoting recovery.

(Example B-11) <Guide for tube feeding> 20 parts by weight of crystalline α-maltose, 1.1 parts by weight of glycine, 0.18 parts by weight of sodium glutamate, 1.2 parts by weight of sodium chloride, 1 part of citric acid Parts by weight, calcium lactate 0.4 parts by weight, magnesium carbonate 0.1 parts by weight,
A formulation comprising 0.01 part by weight of α-glycosyl naringin, 0.01 part by weight of thiamine and 0.01 part by weight of riboflavin obtained by the method of Example A-3 is prepared. Each 24 g of the blend was filled into a laminated aluminum pouch and heat sealed to prepare a tube nutritional supplement.

The present tube-feeding nutrient can be used for a bag of about 300 to 50
It is dissolved in 0 mL of water and can be advantageously used as an oral or parenteral nutritional supplement for the nasal cavity, stomach, intestine, etc. by the tube method.

(Example B-12) <Bath agent> 21 parts by weight of DL-sodium lactate, 8 parts by weight of sodium pyruvate, α- obtained by the method of Example A-1
5 parts by weight of glycosyl naringin and 40 parts by weight of ethanol were mixed with 26 parts by weight of purified water and an appropriate amount of a coloring agent and a flavor to prepare a bath agent.

The product is suitable as a skin-beautifying agent and a skin-whitening agent, and may be used after diluting it 100 to 10,000 times in hot water for bathing. This product is similar to the case of hot water for bathing,
It can also be advantageously used by diluting it for use in face wash water, lotion and the like.

Example B-13 <Emulsion> 0.5 parts by weight of polyoxyethylene behenyl ether, 1 part by weight of polyoxyethylene sorbitol tetraoleate, 1 part by weight of lipophilic glyceryl monostearate, 0.1 part of pyruvic acid 5 parts by weight, behenyl alcohol 0.
5 parts by weight, avocado oil 1 part by weight, α-glycosyl naringin 1 part by weight obtained by the method of Example A-3, vitamin E
And an appropriate amount of a preservative are dissolved by heating according to a conventional method, and 1 part by weight of sodium L-lactate, 5 parts by weight of 1,3-butylene glycol, 0.1 part by weight of carboxyvinyl polymer and 85.3 parts by weight of purified water are dissolved therein. Was added to the mixture, and the mixture was emulsified with a homogenizer. Further, an appropriate amount of a flavor was added, followed by stirring and mixing to produce an emulsion.

The product can be advantageously used as a sunscreen, a skin beautifying agent, a fairing agent and the like.

Example B-14 <Cream> 2 parts by weight of polyoxyethylene glycol monostearate, 5 parts by weight of self-emulsifying glyceryl monostearate, α-glucosyl naringin 2 obtained by the method of Example A-2 Parts by weight, 1 part by weight of liquid paraffin, 10 parts by weight of glyceryl trioctanoate and an appropriate amount of preservative
Heat and dissolve according to a conventional method, and add 2 parts by weight of L-lactic acid,
5 parts by weight of 1,3-butylene glycol and 66 parts by weight of purified water were added, and the mixture was emulsified by a homogenizer, and an appropriate amount of a flavor was further added, followed by stirring and mixing to produce a cream.

The product can be advantageously used as a sunscreen, a skin beautifying agent, a fairing agent and the like.

[0094]

As described in the text, naringin, a novel substance of the present invention, has D-glucose residues in equimolar or more than α.
The bound α-glycosyl naringin is excellent in water solubility, has a bitter taste, is not toxic, and is easily hydrolyzed to naringin and D-glucose in vivo, thereby exhibiting the original physiological activity of naringin .

The α-glycosyl naringin can be easily produced by a biochemical method in which a solution containing naringin and an α-glucosyl sugar compound is reacted with a glycosyltransferase. It is easy to implement.

Further, it has been found that the reaction can be carried out with an increased concentration of naringin, and that α-glycosylnaringin can be easily produced at a high concentration. In addition, when the reaction solution is purified, the reaction solution is made porous. It has also been found that α-glycosyl naringin can be purified by contact with a synthetic adsorbent, which greatly facilitates mass production of α-glycosyl naringin.

The α-glycosyl naringin thus obtained has characteristics such as good water solubility, good light fastness and stability, and a sufficient physiological activity, and is a highly safe natural type. Not only as a vitamin P enhancer, but also as a bitter, antioxidant, stabilizer, quality improver,
As a preventive agent, a therapeutic agent, an ultraviolet absorber, an anti-deterioration agent, etc., it can be used for various compositions such as foods and drinks, foods, feed, feed, anti-sensitivity agents, skin-skin agents, skin-whitening agents, and plastic products. It is advantageously used for applications.

Therefore, the industrial production method of α-glycosyl naringin according to the present invention and the establishment of its use can be found in food and drink,
It has great industrial significance in the cosmetics, pharmaceuticals and plastic industries.

[Brief description of the drawings]

FIG. 1 is an infrared absorption spectrum of α-glycosyl naringin sample [II] as an example of the present invention.

FIG. 2 is an infrared absorption spectrum of naringin as a control.

Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61P 43/00 105 A61P 43/00 105 // C07H 17/07 C07H 17/07 C12N 9/24 C12N 9/24 C12P 19/60 C12P 19 / 60 (58) Fields investigated (Int.Cl. ⁷ , DB name) A61K 31/7048 A23L 1/03 A61K 7/00 A61K 47/26 A61P 17/00 A61P 43/00 C07H 17/07 C12N 9/24 C12P 19/60 CA (STN) EMBASE (STN) MEDLINE (STN)

Claims (15)

(57) [Claims] 1. A high-concentration suspension having a concentration of 1 w / v% or more.
Concentration of 1 g dissolved in carrot or alkaline pH
/ V% or higher concentration solution naringin, α-glucosi
Α-glucosidase, in a solution containing a sucrose compound,
Cyclomaltodextrin glucanotransferr
Acts on glycosyltransferases selected from the group consisting of
An anti-sensitive disease agent obtained by adding α-glycosyl naringin in which D-glucose residues are α-bonded to naringin in equimolar or more. 2. α-Glycosyl naringin reacts with glycosyltransferase, then with amylase,
Α-glycosyl obtained by forming glucosyl naringin and / or α-maltosyl naringin
2. The anti-sensitive disease agent according to claim 1, wherein the agent is naringin. 3. The α-glycosyl naringin obtained by contacting a solution containing the generated α-glycosyl naringin with a porous synthetic adsorbent and purifying the solution, wherein the α-glycosyl naringin is obtained. Or the anti-sensitive disease agent according to 2. 4. The anti-sensitive disease agent according to claim 1, wherein the α-glycosyl naringin is α-glycosyl naringin obtained by being produced in the presence of an antioxidant. 5. A method according to claim 5, wherein said compound is α-glycosyl naringin.
Thiamine, riboflavin, vitamin C and vitamin E
The anti-sensitive disease agent according to any one of claims 1 to 4, comprising a vitamin selected from the group consisting of: 6. A high-concentration suspension of naringin having a concentration of 1 w / v% or more, or a concentration of 1 w / v% dissolved in alkaline pH.
/ V% or more of a high-concentration solution of naringin and an α-glucosyl sugar compound in a solution containing α-glucosidase,
A stabilizer obtained by reacting a glycosyltransferase selected from cyclomaltodextrin glucanotransferase and α-amylase, wherein the stabilizer contains α-glycosyl naringin in which D-glucose residues are α-bonded to equimolar or more to naringin. 7. α-Glycosyl naringin reacts with glycosyltransferase, then reacts with amylase,
Α-glycosyl obtained by forming glucosyl naringin and / or α-maltosyl naringin
The stabilizer according to claim 6, which is naringin. 8. The α-glycosyl naringin obtained by contacting a solution containing the produced α-glycosyl naringin with a porous synthetic adsorbent and purifying the resulting solution, wherein the α-glycosyl naringin is obtained. Or the stabilizer according to 7. 9. The stabilizer according to claim 6, wherein the α-glycosyl naringin is α-glycosyl naringin obtained by being produced in the presence of an antioxidant. 10. The stabilizer according to claim 6, further comprising a vitamin selected from thiamine, riboflavin, vitamin C and vitamin E, together with α-glycosyl naringin. 11. A highly concentrated suspension of naringin having a concentration of 1 w / v% or more, or a concentration of 1 dissolved in alkaline pH.
A solution containing w / v% or more of a high-concentration solution of naringin and an α-glucosyl sugar compound is reacted with a glycosyltransferase selected from α-glucosidase, cyclomaltodextrin glucanotransferase and α-amylase. And α-glycosyl naringin in which D-glucose residues are α-bonded to naringin in equimolar or more. 12. α-Glycosyl naringin reacts with glycosyltransferase, then reacts with amylase,
The cosmetic according to claim 11, wherein the cosmetic is α-glycosyl naringin obtained by producing -glucosyl naringin and / or α-maltosyl naringin. 13. The method according to claim 11, wherein the α-glycosyl naringin is α-glycosyl naringin obtained by contacting a solution containing the produced α-glycosyl naringin with a porous synthetic adsorbent and purifying the solution. Or the cosmetic according to 12. 14. An α-glycosyl obtained by producing α-glycosyl naringin in the presence of an antioxidant.
14. Naringin, characterized in that it is naringin
Cosmetics as described. 15. The cosmetic according to claim 11, further comprising a vitamin selected from thiamine, riboflavin, vitamin C and vitamin E, together with α-glycosyl naringin.