JP2008125515A - Method for producing germination differentiated plant seed, food containing easily water-soluble polyphenol and/or antioxidation substance of germination differentiated plant seed, and apparatus for germination differentiation of plant seed - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a germination differentiated seed, having increased contents of easily water-soluble polyphenols and antioxidation substances, food containing easily water-soluble polyphenols and/or antioxidation substances of the germination differentiation plant seed, and to provide a method for producing the food. <P>SOLUTION: The germination of a seed is induced within a selected temperature range. The germination induction is carried out, in an atmosphere having elevated carbon dioxide concentration and/or a reduced oxygen concentration. The easily water-soluble polyphenols can be extracted from an aqueous solution, without having to use organic solvent, such as alcohol, by irradiating the seed to light. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  TECHNICAL FIELD The present invention relates to germinated and differentiated seeds with increased readily water-soluble polyphenols and antioxidants, germinated and differentiated plant seeds containing easily water-soluble polyphenols and / or antioxidants, and a method for producing the same. According to the present invention, it is possible to produce germinated and differentiated seeds containing a large amount of easily water-soluble polyphenols useful as food ingredients, cosmetic ingredients or pharmaceutical ingredients, and easily extract readily water-soluble polyphenols from the seeds. Can do.

The polyphenol contained in a plant body is a general term for plant components having a phenolic hydroxy group in the molecule, and is contained in almost all plants, and the number thereof reaches 5,000 or more. Polyphenols are broadly classified into flavonoids, phenolic acids, ellagic acids, lignans, curcumins, coumarins, procyanines, caffeic acids, and various physiological activities such as radical scavenging ability and antioxidant activity. have. Specific bioactive functions have been reported to have functions such as antimutagenic action, anticancer action, suppression of blood pressure increase, antidiabetic action, and antiallergy (Wine chemistry Yair Margalit The Wine Appreciation Guide). 1997 Ca San Francisco). For this reason, taking an appropriate amount of polyphenols is considered to be good for health, and has been conventionally used in many foods, health foods, pharmaceuticals and cosmetics.
JP 2001-158739 A

Some of these polyphenols are present in plants as water-soluble polyphenols, but many are included in plants as poorly water-soluble aglycones. In particular, polyphenols contained in plant seeds are mostly poorly water-soluble polyphenols, and when extracting such polyphenols from plants, it is difficult to extract with a buffer solution that does not contain water or alcohol. In addition, an extract containing alcohol had to be used as described above. Further, since a poorly water-soluble polyphenol cannot be used in the state of an aqueous solution, its range of use tended to be relatively limited. For example, when it is used as a cosmetic, it can be contained in a cream, but it is not contained in a lotion free of alcohol. Furthermore, even when contained in health foods orally and used orally, generally, poorly water-soluble polyphenols are less likely to be absorbed as compared to readily water-soluble polyphenols, so that it has been considered that the effect cannot be sufficiently exerted. Therefore, even if polyphenols are actually contained in food or the like, if they are poorly water-soluble polyphenols, it is considered that sufficient effects cannot be obtained due to poor absorption.
Therefore, there is a need for a method and a production method that can produce germinated and differentiated seeds containing a large amount of useful water-soluble polyphenols useful for health foods, cosmetics, and the like, and can easily extract water-soluble polyphenols from the seeds.

The present invention performs germination plant seed retention under atmospheric conditions of carbon dioxide concentration of substantially 2000 ppm or more and / or oxygen concentration of substantially 18% by volume or less, and retention under germination temperature conditions. This is a method for producing germinated and differentiated seeds with increased readily water-soluble polyphenols. Plant seeds are hydrated by spraying water, sprinkling water, or other methods. The hydration is performed before the germination induction step and / or during germination. Furthermore, before the said germination induction | guidance | derivation process, it is the manufacturing method of the germination differentiation seed which the water-soluble polyphenol increased including the process of heating to the seed of a plant. The said germination induction process is a germination induction process which further irradiates light. Specifically, the light irradiation is preferably 5000 lux or more per day and 2 hours or more.
In the germination induction step, it is preferable that the carbon dioxide concentration in the atmospheric condition is 2,000 ppm or more and 50,000 ppm or less and is intermittently maintained for 60% or more of the germination induction step time.

In the germination induction step, it is preferable that the oxygen concentration in the atmospheric condition is 4% by volume or more and 18% by volume or less, and is intermittently maintained for 60% or more of the germination induction step time. In the germination induction step, the germination temperature is preferably 20 ° C to 45 ° C. The heating temperature is preferably 5 ° C. or higher and 30 ° C. or lower, which is the median temperature for germination. The germination induction step is a germination induction step performed by depositing seeds, and it is preferable to perform retention under the germination temperature condition by utilizing a temperature increase due to germination heat and a temperature decrease due to watering.
The present invention provides the seeds of germinable plants with a carbon dioxide concentration of 2000 ppm or more and / or an oxygen concentration of 18% by volume or less in an atmospheric condition, and a temperature condition of 20 ° C. to 45 ° C. The present invention relates to a food material containing a readily water-soluble polyphenol and / or antioxidant obtained by holding. Plant seeds are a germination induction process in which water is contained and / or light is irradiated. The light irradiation is preferably 5000 lux or more per day and 2 hours or more. It is preferable that the heating temperature is maintained at 5 ° C. or higher and 30 ° C. or lower, which is the median temperature for germination.

The food contains an easily water-soluble extract derived from plant seeds containing polyphenols. The food includes plant seeds. The plant seeds contained in the food preferably contain at least 1 milligram of a readily water-soluble polyphenol mixture per gram of plant seeds. Preferably, it contains at least 2 milligrams of readily water-soluble polyphenol per gram of plant seed. Furthermore, plant seeds contain an antioxidant equivalent to at least 40 μmol DPPH per gram of plant seeds, have a water-soluble polyphenol content of at least 8 times that of ungerminated seeds, and are basically grape plant seeds. Preferably it consists of.
As another aspect of the present invention, there is an apparatus for germinating plant seeds having an increased amount of water-soluble polyphenol. This apparatus has means for holding at atmospheric conditions of carbon dioxide concentration of substantially 2000 ppm or more and / or oxygen concentration of substantially 18% by volume or less. The apparatus further has means for keeping the plant seeds within the germination temperature range and means for hydrating the plant seeds. Furthermore, it is preferable that this apparatus has a light source that irradiates the plant seed with light at 5000 lux or more for 2 hours or more per day.

The present invention relates to a system and method for increasing the amount of water-soluble polyphenols in germinated and differentiated seeds. According to the examples, germinable plant seeds are maintained under atmospheric conditions of carbon dioxide concentration of substantially 2000 ppm or higher and / or oxygen concentration of substantially 18% by volume or lower, holding at germination temperature conditions, and Systems and methods are provided for inducing the germination process by performing watering.
The first, second, etc. terms are used to describe the various elements, but this is to distinguish the elements and not to limit them. For example, the first element may be expressed as the second element, and similarly the second element may be expressed as the first element, but this does not depart from the scope of the invention. The term “and / or” includes any and all of the listed items, alone or in combination.

In the following detailed description, reference is made to the drawings that accompany the specific examples. The foregoing drawings are illustrative of the invention as a specific example and are not intended to be limiting. These implementations have been described in detail and enable one skilled in the art to do so. Various other embodiments and implementations are also consistent with the principles of the present invention. Other embodiments and implementations may be used, and structural changes and / or substitutions may be made to various elements without departing from the spirit and scope of the present invention. The following detailed description is, therefore, not to be construed in a narrow sense.
A polyphenol is a plant-derived compound having two or more phenolic hydroxy groups in the same molecule. Flavonoids, phenylcarboxylic acids, ellagic acids, lignans, curcumins, coumarins, procyanines, coffee There are 5,000 or more types of polyphenols, which are divided into acid types. Examples include catechin, tannin, anthocyanin, proanthocyanin, quercetin, isoflavone, rutin, chlorogenic acid, cacao mass polyphenol, and xanthone. Plant seeds containing these polyphenols include teas, apples, blueberries, lotus roots, etc. as seeds containing a large amount of catechins, and teas, oysters, bananas, grapes, etc. as seeds containing a large amount of tannins. The seeds rich in proanthocyanins are grapes, apples, blueberries, etc. The seeds rich in quercetin are onions, spinach, broccoli, spring chrysanthemum, and the seeds rich in isoflavones are soybeans, clover. As seeds rich in rutin, buckwheat, potato, prunes, spring chrysanthemum, etc. as seeds rich in chlorogenic acid, cacao, coffee, etc. as seeds rich in cocoa mass polyphenols, xanthone With high seed content and Te, such as Mangosteen, and the like.

As a result of intensive research on a method for stably obtaining a water-soluble polyphenol from a plant body, the present inventors have found that when a plant seed germinates, the water-soluble polyphenol increases. (US patent application 11 / 246,442). Furthermore, by adjusting the oxygen concentration and / or carbon dioxide concentration and germination temperature at the time of heat treatment before germination induction and germination differentiation induction, the amount of water-soluble polyphenols contained in germinated and differentiated seeds is remarkably increased. Found to increase. The present invention is based on these findings. In particular, the amount of water-soluble polyphenols in germinated and differentiated seeds is increased by germinating and seeding seeds under atmospheric conditions of carbon dioxide concentration of 2000 ppm or more and / or oxygen concentration of substantially 18% by volume or less. I found it.
The easily water-soluble polyphenol in the present specification is not particularly limited as long as it is a polyphenol that is easily extracted with a buffer solution or water that does not contain an organic solvent such as alcohol. For example, most of glycoside polyphenols obtained by adding sugar to poorly water-soluble polyphenols, which are aglycones contained in large amounts in plant seeds, are readily water-soluble polyphenols. Specifically, glycoside flavonoids such as rutin, daidzin, naringin and hesperidin are readily water-soluble polyphenols. A typical polyphenol, anthocyanidin, is sparingly water-soluble, but anthocyanin, which is a glycoside having anthocyanidin as an aglycone, is a water-soluble polyphenol. In addition, glycoside proanthocyanidins, which are glycosides of proanthocyanidins contained in a large amount in grape seeds, are also readily water-soluble polyphenols.

  The seeds of the plant used in the method of the present invention are not particularly limited as long as the easily water-soluble polyphenol is increased by germination and differentiation. Many seeds contain poorly water-soluble polyphenols, which are fat-soluble aglycones. These aglycones become easily water-soluble polyphenols by becoming glycosides to which sugar is added. Seeds that increase water-soluble polyphenols by germination and differentiation include vines such as grapes, asteraceae sunflowers, safflowers, and burdock, and legumes such as clover, peanuts, and fenugreek. Family) such as sesame, sesame, sesame, linseed, perilla, rosemary, thyme, sage, and mint, soda buckwheat, rice, corn, wheat, wild rice, American red cherries in the family Rosaceae such as barley, millet, and millet, apricots, almonds, plums, strawberries, and loquats, grapefruits in citrus, oranges, St. John's wort in Hypericaceae, and eggplants such as mangosteen Tomatoes and green peppers, such as carrots from the celery family, Gore, such as pomegranate Department of pomegranate, polygonaceae of Japanese knotweed, such as cocoa of Sterculiaceae, trifoliate orange family, the ginkgo family, mention may be made of mulberry family, polygonaceae, camellia family, Oleaceae, and holly Department of seeds, and the like.

The reason why the water-soluble polyphenol is increased to a high concentration in the plant seed by the method of the present invention is not completely elucidated, but can be inferred as follows. However, the present invention is not limited by the following description.
The poorly water-soluble polyphenols in the plant seeds are converted into easily water-soluble polyphenols such as glycoside polyphenols by germination differentiation. This conversion of poorly water-soluble polyphenols to glycoside polyphenols is thought to occur via enzymatic reactions, etc., but as germination and differentiation proceed, accumulated water-soluble polyphenols are used for germination and differentiation. To do. Easily water-soluble polyphenols are thought to be consumed by seed respiration. By controlling the carbon dioxide concentration and / or the oxygen concentration, it is considered that the respiration of the seed can be suppressed, the consumption of the water-soluble polyphenol can be suppressed, and the water-soluble polyphenol concentration in the seed can be increased.
However, the action of the present invention not only suppresses the respiratory action, but also lowers the pH in the seed by lowering the carbon dioxide concentration, activates the enzyme reaction in the seed by changing the germination temperature, and by light irradiation. The influence and the like are also considered to affect the conversion of the poorly water-soluble polyphenol to the glycoside polyphenol and the accumulation of the water-soluble polyphenol. Of course, these hypotheses do not limit embodiments, nor are they dependent on or limited to any particular hypothesis or theory relating to the device.

The seed of the plant used for the method for increasing the readily water-soluble polyphenol of the present invention is a germinable seed. Germinated plant seeds are seeds that are not "dormant". For example, plant seeds with a moisture content of 9-12% are considered dormant.
The heating of the method of the present invention can be performed simultaneously with water as described above, but can also be performed as a single step. In order to change the seeds of a plant from a dormant state to a germinable state, specifically, the seeds can be brought into a germinable state by heating or containing water. However, without performing any special pretreatment, the water content of the seeds can be increased by watering during the germination induction process, so that the seeds can be germinated.
It is particularly effective to combine heating and hydration. Seeds that have become germinable by combining heating and water content generally have a higher water-soluble polyphenol content than seeds that can be germinated only by heating or water content. Heating also promotes germination. Furthermore, seeds can be made to germinate only with water without heating, but the germination rate increases by simultaneously heating the seeds and water. When heating and hydration are simultaneously performed, for example, the seed can be immersed in heated water for a certain period of time.
Even if the seed is not subjected to special pretreatment, it can be germinated by spraying water within the range of germinable temperature.
Plant seeds are allowed to germinate by heating in the range of germinable temperature, and the amount of polyphenol increases. In this embodiment, the temperature of the seed is maintained at a temperature that is not less than the appropriate temperature for germination and does not denature the protein in the plant seed. It is preferable to heat at a temperature 5 to 30 ° C. higher than the median temperature of each germination.
The heating temperature for heating the plant seeds is preferably in a temperature range higher than the plant germination optimum temperature. Although the optimal temperature for germination of plant seeds varies depending on the plant, it is 10 ° C to 45 ° C, preferably 20 ° C to 40 ° C. For example, the optimal temperature for germination of grape seeds is 20 ° C to 35 ° C, the optimal temperature for germination of mungbean is 20 ° C to 35 ° C, the optimal temperature for germination of radish is 18 ° C to 30 ° C, and the optimal temperature for germination of soybean is 10 ° C to 30 ° C. . The median appropriate temperature for germination of grapes and mungbean is about 28 ° C., the median optimal temperature for germination of radish is 24 ° C., and the median optimal temperature for germination of soybean is 20 ° C.

The higher the temperature, the more easily the water-soluble polyphenol. This is thought to be related to certain proteins in plant seeds. These proteins are thought to have an affinity for polyphenols. Since there is a competitive reaction with polyphenols, it is expected to reduce the amount of polyphenols required for the desired readily water-soluble materials.
An increase in temperature within 5 ° C. from the optimum germination temperature has little effect on the germination differentiation reaction that promotes the production of water-soluble polyphenols. However, it was confirmed that the amount of readily water-soluble polyphenols increased even with a slight temperature increase of about 5 ° C. One hypothesis is that this increase in production is partly due to promoting protein denaturation. It is thought that the decrease in the amount of protein hinders or suppresses a competitive reaction that reduces the amount of water-soluble polyphenol produced. While this may explain the effects of water content and temperature of the disclosed process, the processes, principles and examples disclosed herein are not dependent on this description or other theory.
A heating temperature 5 to 20 ° C. higher than the median temperature for germination was selected. Specifically, it is preferable to heat at 33 ° C. to 48 ° C. for grapes and mungbeans, and at 25 ° C. to 50 ° C. for soybeans.
The water content in the production method of the present invention can be performed simultaneously with the heating, but can also be performed alone. Furthermore, in the germination differentiation process described later, it is possible to hydrate the plant seeds by watering. This is because the grape seeds shift from the dormant state to the germination and differentiation state by the heating, and the plant seeds absorb a large amount of water with the start of germination and differentiation.

It is known that the amount of water-soluble polyphenol contained in germinated and differentiated seeds is affected by the carbon dioxide concentration and / or the oxygen concentration in the atmosphere. The increase in the carbon dioxide concentration and the decrease in the oxygen concentration increase the production of water-soluble polyphenols in germinated and differentiated seeds. However, adjusting both the carbon dioxide concentration and the oxygen concentration yields more readily water-soluble polyphenols (eg, increasing carbon dioxide and decreasing oxygen). Further, when the germination temperature is adjusted, the seed is sprinkled, and the seed is irradiated with light, the amount of the water-soluble polyphenol further increases.
The oxygen concentration is lower than the atmospheric oxygen concentration (20%), and is not particularly important as long as it increases the water-soluble polyphenol of germinated and differentiated seeds. The oxygen concentration is preferably 4% to 20% by volume, but 4% to 18%, 4% to 15% by volume is also effective. If it is less than 4% by volume, oxygen respiration of seeds is weak and the amount of easily water-soluble polyphenols decreases.
Heretofore, it has been postulated that the consumption of readily water-soluble polyphenols in seeds is suppressed when the oxygen concentration decreases. However, this embodiment does not depend on this or any other theory, and is not limited by any theory.
The oxygen concentration can be controlled by using an incubator, desiccator, cultivation room or the like that can adjust the oxygen concentration. However, oxygen is consumed by the respiration of germinated and differentiated seeds in a sealed or poorly ventilated device. For example, by depositing several layers of plant seeds in a sealable container, the oxygen concentration is reduced and adjusted. It is also possible. Oxygen in the container is consumed by seeds that have started germination and differentiation, and the oxygen concentration decreases. In order to prevent the oxygen concentration range from becoming lower than the concentration range where the effect is obtained, the oxygen concentration can be maintained at an appropriate concentration by replacing it with the normal atmospheric atmosphere at regular intervals. The replacement with the normal atmosphere can be performed by replacing the gas between the deposited seeds by watering which will be described later.
The oxygen concentration may be maintained at a constant concentration such as 5% by volume, 10% by volume, or 15% by volume in the germination induction step, but substantially maintained at 18% by volume or less. It only has to be done. “Substantial” in the oxygen concentration of the germination induction step means that the oxygen concentration is maintained at a predetermined concentration at 60% or more, preferably 80% or more, more preferably 90% or more of the total time of the germination induction step. To do. The time for which the predetermined concentration is maintained may be continuous, but is preferably maintained intermittently.

The carbon dioxide concentration is higher than the carbon dioxide concentration in the atmosphere, and it is not particularly limited as long as it is a concentration that increases easily water-soluble polyphenols of germinated and differentiated seeds, suppresses consumption in seeds, and accumulates. Preferably from 2,000 ppm (0.2 vol%) to 100,000 ppm (10 vol%), more preferably from 4,000 ppm (0.4 vol%) to 50,000 ppm (5 vol%), most preferably 000 ppm (1.2% by volume) to 20,000 ppm (2% by volume). If the amount is less than 2000 ppm, the respiratory repression effect is not observed, and easily water-soluble polyphenol is consumed, and if it exceeds 10%, germination is impaired.
The carbon dioxide concentration can be controlled by using an incubator, a desiccator, a cultivation room or the like that can adjust the carbon dioxide concentration. However, it is also possible to increase and adjust the carbon dioxide concentration by carbon dioxide discharged by the respiration action of germinated and differentiated seeds without using a special device. For example, by depositing several layers of plant seeds in a sealable container, the carbon dioxide concentration is discharged by the respiration action of seeds that have started germination differentiation, and the carbon dioxide concentration increases. In order to prevent the carbon dioxide concentration range from becoming higher than the concentration range where the effect is obtained, the carbon dioxide concentration can be maintained at an appropriate concentration by replacing air at regular intervals. The air can be replaced by replacing the air between the deposited seeds by watering which will be described later.
The carbon dioxide concentration may be maintained at a constant concentration such as 2,000 ppm, 10,000 ppm, or 50,000 ppm continuously in the germination induction step, but substantially around 12,000 ppm. It only has to be held. “Substantial” in the oxygen concentration of the germination induction step means that the oxygen concentration is maintained at a predetermined concentration at 60% or more, preferably 80% or more, more preferably 90% or more of the total time of the germination induction step. To do. The time for which the predetermined concentration is maintained may be continuous, but is preferably maintained intermittently.

The germination temperature condition in the germination induction process is not limited as long as it is a temperature condition that allows the seeds of the plant to germinate and differentiate. The temperature at which germination can be differentiated varies depending on the plant seed, but for example, it can be carried out at a suitable temperature for germination of the plant. Although the suitable temperature for germination of plant seeds varies depending on the plant seed as described above, it is 10 ° C to 50 ° C, preferably 20 ° C to 45 ° C, more preferably 26 ° C to 42 ° C. In the germination induction step, plant seeds may be maintained at a constant germination temperature, or may be maintained within a constant temperature range. When plant seeds are held within a certain temperature range, they may be held under conditions where the temperature rises and falls repeatedly.
The temperature condition can be controlled by using an incubator capable of adjusting the temperature or by using a cultivation room capable of adjusting the room temperature. The temperature can be maintained by a conventional heater, cooling device, air control, electronic control device, or the like. It is also possible to adjust the germination temperature conditions by the heat of germination due to the respiration of plant seeds during germination and watering.
Various methods may be taken simultaneously to maintain temperature and temperature range. For example, germination temperature conditions can be adjusted by germination heat and watering. By depositing plant seeds in several layers, germination heat is generated by the respiration action of seeds that have started germination differentiation. The germination heat generated because the seeds are deposited does not dissipate, and the temperature between the deposited seeds increases with time. It is possible to adjust the germination temperature within a certain temperature range by lowering this increased temperature by watering performed at regular intervals. When the temperature condition is adjusted by the germination heat and watering as described above, as shown in FIG. 1, the temperature repeatedly increases and decreases gradually within the temperature condition range in which the seeds of the plant can germinate and differentiate.
Even when maintained at a constant temperature, the water-soluble polyphenols in the germinating plant seeds increase sufficiently, but the gradual rise and fall within the temperature condition range can be repeated within the seeds of the water-soluble polyphenols. There is also a possibility that it is affecting the increase.

Watering is performed in order to prevent drying of plant seeds in the germination induction process. “Sprinkling” means spraying water on the seeds, spraying water, intermittently immersing them in water, applying fog to the seeds, and relatively high humidity (at a temperature within the germination temperature range of the seeds). 80% saturation or higher), which means preventing seed drying. Sprinkling plays multiple roles such as maintaining the temperature and temperature range in the germination induction process and maintaining humidity.
Although the temperature of the water to sprinkle is not specifically limited, The temperature within the range of germination temperature conditions is preferable. In particular, when adjusting the temperature condition to a certain range by watering, it is preferable to use water having a relatively low temperature within the germination temperature condition range. The water used for watering preferably contains a bactericidal agent from the viewpoint of hygiene in order to prevent the generation of bacteria and the like in the germination induction process. Examples of the disinfectant include 10 ppm sodium hypochlorite, but other antibacterial compounds may be used.
In the germination induction step, it is preferable to perform light irradiation. Even when light is not irradiated at all, the water-soluble polyphenols in the germinated and differentiated seeds are sufficiently increased. However, when light is irradiated, the water-soluble polyphenols are further increased. For the irradiation of light, a fluorescent tube usually used in plant cultivation can be used. Lights such as three-wavelength type, four-wavelength type, UV, and infrared having different wavelengths may be used singly or in combination. The irradiation time of light is not particularly limited, but is preferably 2 hours or more, more preferably 4 hours or more, most preferably 6 hours or more, and irradiation may be performed for a maximum of 24 hours. Further, light intensity, 20 [mu] mol / m ² · sec ~100μmol / m ² · sec is preferred. As used herein, “germinated and differentiated seed” means a seed that has transitioned from a dormant state to a germinable state. For example, the seed which received the heat shock by the heating process and the moisture content increased by the water-containing process is included. Accordingly, seeds that are not apparently sprouted are also included in the “germinated and differentiated seeds” of the present invention, and seeds that have sprouted and have elongated buds are also included. The germinated and differentiated seed of the present invention contains 1.5 times or more easily water-soluble polyphenol as compared with an ungerminated seed in a dormant state.

A known method can be used as a method for extracting a water-soluble polyphenol from germinated and differentiated seeds containing the water-soluble polyphenol obtained by the method of the present invention. For example, an extract containing an easily water-soluble polyphenol can be obtained by crushing germinated seeds and extracting with a water-based buffer solution that does not contain water or an organic solvent.
As described above, plant seeds are deposited in a sealed or partially sealed container. Furthermore, this structure decreases the oxygen concentration around the seeds and increases the carbon dioxide concentration, thus increasing readily water-soluble polyphenols (antioxidants).
As an apparatus for depositing plant seeds 830 and performing a germination induction process, a germination induction container 800 as shown in FIG. 8 can be exemplified. The germination induction container 800 can be used with the upper surface 810 open, but can also be used in a sealed state. In order to control the carbon dioxide concentration, oxygen concentration, and temperature in the container, it is desirable to use the upper surface 810 and the lower surface 840 in a sealed state. It can also be used in an open state in a cultivation room where the carbon dioxide concentration, oxygen concentration, and temperature can be controlled. Carbon dioxide, oxygen, and temperature in the container are close to the cultivation room.
The germination induction container 800 has a nozzle 860 for spraying water 820 upward. The bottom surface has drain ports (air ports) 850 at intervals of about 1 cm, so that excess water can be drained by watering. The drain is connected to a vertical pipe (not shown) coming out from the bottom of the lower container 840. Each tube is fitted with one arm of a U-shaped tube. The other arm of each U-shaped tube is open to the surrounding environment of the room, and when the water sprinkled accumulates to a certain height of the U-shaped tube, water overflows from the opened arm, and the U-shaped tube is a siphon. Work as. After the sprinkling is finished by this siphon, the inside of the container becomes empty.
According to the method for producing germinated differentiated seeds of the present invention, it is possible to remarkably increase the amount of water-soluble polyphenol contained in germinated differentiated seeds. In addition, according to the germinated and differentiated seed of the present invention, since it contains many readily water-soluble polyphenols compared to ungerminated seeds, extraction and purification of polyphenols from seeds without using an organic solvent such as alcohol Can be easily performed. Furthermore, since the extract containing the water-soluble polyphenol extracted by the method for producing the water-soluble polyphenol of the present invention does not contain a solvent such as alcohol, it can be used in a wide range of applications such as foods, health foods, pharmaceuticals and cosmetics. It can be easily applied.

Hereinafter, the present invention will be described in detail with reference to Examples and Comparative Examples, but these do not limit the scope of the present invention.
For these examples, the amount of water-soluble polyphenols in ungerminated and differentiated seeds was measured by a standard method (Japan Biotechnology Experiment Paper “Biotechnology Experiment” Culture Style Hall). The seeds were crushed, and easily water-soluble polyphenols were extracted with a buffer solution containing no water or organic solvent. The amount of easily water-soluble polyphenol in the solution was measured using light absorption with gallic acid as a standard. Polyphenols and glycosides were measured by the foreign / thiocult method and the phenol / sulfuric acid method, respectively. The compound in the extract was confirmed to be a readily water-soluble glycoside by a method using thin layer chromatography. The antioxidant activity of the water-soluble polyphenols in the extract from the germinated seeds was determined by referring to the Trolox (Trolox: 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) standard. Measured by phenicrylhydrazyl (DPPH) radical scavenging activity.

Example 1: Increase in water-soluble polyphenols in grape seeds After 225 g of grape seeds were washed with running water, they were immersed in an aqueous solution containing 100 ppm of sodium hypochlorite as a fungicide for 1 hour. The seeds were taken out and the 100 ppm sodium hypochlorite solution was exchanged and immersed for 4 hours at 40 ° C. Next, after washing the seeds with distilled water, the water was removed with a soft paper. The seeds were stacked in a germination induction container 800 with a thickness of about 5 cm (FIG. 8). The upper surface of the germination induction container 800 was sealed and placed in a cultivation room at room temperature of 28 ° C., and a 10 ppm sodium hypochlorite solution at 28 ° C. was sprinkled every 4 hours. Approximately 100 germinated seeds (4.12 g) were collected every 24 hours (1 day), 48 hours (2 days), 96 hours (4 days), and 168 hours (7 days).
4.12 g of the collected germinated differentiated seeds were placed in a mortar, 10 ml of distilled water was added, and the mixture was crushed with a pestle. The crushed material was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
The polyphenol contained in the extract obtained from germinated differentiated grape seeds was analyzed by the method described above. In one method, the extract contained 302 μg polyphenols. According to another glycoside analysis, the presence of 366 μg glycoside was confirmed. That is, a large amount of water-soluble glycoside polyphenol was present in the extract. Proanthocyanidin is the most abundant polyphenol contained in grape seeds, and the glycoside polyphenol contained in the germinated seed is considered to be a glycoside polyphenol obtained by adding sugar to proanthocyanidin or a derivative thereof.

Example 2: Extraction of readily water-soluble polyphenols from ungerminated seeds About 100 grape seeds (Riesling species: moisture content 9.5 to 11.0) (4,12 g) derived from juice pomace were placed in a mortar and distilled. 10 ml of water was added and crushed with a pestle. This was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
Compared with the extract obtained from the germinated differentiated grape seed of 24 hours after Example 1, about half of the water-soluble polyphenol was contained.

Example 3 Effect of Oxygen Concentration and Carbon Dioxide Concentration After 225 g of mungbean seed, clover seed, sunflower seed, or yuzu seed were washed with running water, an aqueous solution containing 100 ppm of sodium hypochlorite as a bactericide was used for 4 hours. Soaked. The seeds were then washed with distilled water and the water was removed with a soft paper. The seeds were stacked in a germination induction container 800 with a thickness of about 5 cm (FIG. 8).
The germination induction container 800 was installed in a cultivation room at room temperature (28 ° C.), and a 10 ppm sodium hypochlorite solution at 28 ° C. was sprinkled every 4 hours to induce germination for 48 hours, and about 675 g of germinated differentiated seeds were collected.
4.12 g of the collected germinated differentiated seeds were put in a mortar, 10 ml of distilled water was added and crushed with a pestle. This crushed material was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
The temperature change in the warm germination induction process of mungbean seeds using the germination induction container 800 was measured from the start of germination induction to 24 hours. The temperature is a temperature at a position 3 cm from the surface where the seeds are stacked. Although the cultivation room is 28 ° C., the temperature in the germination induction container gradually increases with the heat of germination of mungbean seeds. Although it reaches 39 ° C. in 4 hours from the start of germination induction, a 10 ppm sodium hypochlorite aqueous solution with a water temperature of 28 ° C. is sprinkled to lower the temperature of the germinated product to 28 ° C. (FIG. 1). This temperature was measured 3 cm from the surface, but the inside of the stacked seeds showed almost the same temperature change.
Changes in oxygen concentration in the warm germination induction process of mungbean seeds using the germination induction container 800 were measured from the start of germination induction to 24 hours. The oxygen concentration is a concentration at a position within 3 cm from the surface where the seeds are stacked. Since it is replaced with the outside air every 4 hours, the oxygen concentration in the normal atmospheric atmosphere temporarily temporarily. However, oxygen is consumed immediately by the respiration action of the seed and falls to 15% or less. (Figure 3). This concentration was measured within 3 cm from the surface, but the inside of the stacked seeds showed almost the same concentration change.
Changes in carbon dioxide concentration in the germination induction process of mungbean seeds using the germination induction container 800 were measured from the start of germination induction to 24 hours. The carbon dioxide concentration is a concentration at a position within 3 cm from the surface where the seeds are stacked. Since it is replaced with the outside air every 4 hours, the carbon dioxide concentration in the normal atmosphere is temporarily increased, but carbon dioxide immediately increases due to the respiration action of the seeds. After sprinkling, carbon dioxide rapidly reaches 8000 ppm (FIG. 5). The carbon dioxide concentration was measured 3 cm inside from the surface, but the inside of the stacked seeds showed almost the same concentration.

Example 4: Effect of oxygen concentration and carbon dioxide concentration After washing 225 g of mungbean seed, clover seed, sunflower seed or yuzu seed with running water, an aqueous solution containing 100 ppm of sodium hypochlorite as a bactericide was used for 4 hours. It was immersed for a period of time, and heating and water content were performed simultaneously. Next, after washing the seeds with distilled water, the water was removed with a soft paper. Seeds were spread further in the germination container 700 (FIG. 7).
The germination container 700 has an open upper surface 710 and has a nozzle 760 for watering above. There are drainage ports (air ports) 740 at 1 cm intervals on the bottom surface, and excess water can be drained. Since the upper surface is in an open state and has many drainage ports (air ports), the oxygen concentration and carbon dioxide concentration in the container are usually kept almost the same as the concentration in the atmospheric air. At the first time, the germination container 700 was placed in a 38 ° C. cultivation room, and a 10 ppm sodium hypochlorite solution at 36 ° C. was sprinkled every 4 hours. In the second time, the container 700 was placed in a cultivation room at a room temperature of 28 ° C., and a 10 ppm sodium hypochlorite solution at 27 ° C. was sprinkled every 4 hours. After about 48 hours, about 675 g of germinated seeds were collected.
4.12 g of the collected germinated differentiated seeds were put in a mortar, 10 ml of distilled water was added and crushed with a pestle. This crushed material was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
As shown in FIG. 2, the temperature change in the germination induction process of mungbean seeds using the germination container 700 was measured from the start of germination induction to 24 hours. The temperature was measured on the surface of the seed that had been spread. When the cultivation room was 38 ° C., the temperature dropped to 36 ° C. by watering a 10 ppm sodium hypochlorite aqueous solution. Therefore, as shown by the curve 210 in FIG. 2, the temperature is maintained between 36 ° C. and 38 ° C. in the first test. When the cultivation room was 28 ° C., the temperature dropped to 27 ° C. by watering a 10 ppm sodium hypochlorite aqueous solution. Therefore, as shown by the curve 220 in FIG. 2, the temperature is kept between 27 ° C. and 28 ° C. in the second test. Oxygen concentration change in the germination induction process of mungbean seeds using the germination container 700 is induced to germinate. Measurements were taken up to 24 hours from the start. As for the oxygen concentration, the concentration of the surface of the seeds spread in one layer was measured. The oxygen concentration was approximately 21% throughout the germination induction process (FIG. 4).
Changes in carbon dioxide concentration in the germination induction process of mungbean seeds using the germination container 700 were measured from the start of germination induction to 24 hours. The carbon dioxide concentration is the concentration on the surface of the seed that has been spread further. The carbon dioxide concentration was approximately 300 ppm throughout the germination induction process (FIG. 6).

Extract analysis
Extracts obtained from mungbean seeds, clover seeds, sunflower seeds and yuzu seeds were analyzed by the method described above.
As shown in Table 1, germinated and differentiated seeds are at least 11 times higher than mung bean seeds, at least 5 times higher than clover, at least 4.1 times higher than sunflower, and at least 20 times higher than sunflower. The above readily water-soluble polyphenol was contained. On the other hand, the germinated seeds of Example 4 had less water-soluble polyphenols than germinated and differentiated seeds 3 of Examples even when the cultivation room temperature was 38 ° C. or 28 ° C. (Table 1).

Example 5: Effect of temperature
After washing 225 g of mungbean seeds, clover seeds, sunflower seeds and grape seeds with running water, germination optimum temperature (median value) or an aqueous solution containing 100 ppm sodium hypochlorite at a temperature 10 ° C higher than germination optimum temperature (median value) for 4 hours Immersion was performed, and heating and water content were performed simultaneously. The seeds were then washed with distilled water and the water was removed with a soft paper. The seeds were stacked in a germination induction container 800 with a thickness of about 5 cm (FIG. 8). A germination induction container 800 was installed in a cultivation room at room temperature (28 ° C.), and a 10 ppm sodium hypochlorite solution at 28 ° C. was sprinkled every 4 hours to induce germination for 8 hours, and about 675 g of germinated differentiated seeds were collected. The median appropriate temperature for germination of mung bean seeds is 28 ° C., the median optimal temperature for germination of clover seeds is 24 ° C., the median optimal temperature for germination of sunflower seeds is 24 ° C., and the median appropriate temperature for germination of grape seeds is 28 ° C. .
4.12 g of germinated differentiated seeds collected from mungbean seeds, clover seeds, sunflower seeds, or grape seeds were placed in a mortar, and 10 ml of distilled water was added and crushed with a pestle. This was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
The polyphenol content of each sample was analyzed by the method described above. The analysis results are shown in Table 2. When the heating step was carried out at a temperature 10 ° C. higher than the median appropriate germination temperature, the increase in water-soluble polyphenols was more marked than when the heating step was carried out at an average germination optimum temperature. In mungbean seeds, even when the heating step was performed at temperatures of 5 ° C., 20 ° C., and 30 ° C. higher than the appropriate temperature for germination, the increase in water-soluble polyphenols was more significant than when the heating step was performed at the average appropriate temperature for germination.

Example 6: Effect of Oxygen Concentration The effect of a decrease in oxygen concentration on the increase of readily water-soluble polyphenols during the germination induction process was examined using grape seeds. 225 g of grape seeds were washed with running water, and then immersed in a 100 ppm sodium hypochlorite solution at 40 ° C. for 4 hours.
The seeds were then washed with distilled water and the water was removed with a soft paper. The seeds were put in a desiccator and a gas whose oxygen concentration was adjusted to 5%, 10% and 15% by mixing nitrogen gas and air was aerated. Usually, the oxygen concentration in the atmosphere is about 21% by volume. The carbon dioxide concentration in the desiccator was almost 350 ppm under all conditions. The temperature was maintained at 28 ° C., and a 10 ppm sodium hypochlorite solution at 28 ° C. was sprinkled every 4 hours to induce germination for 48 hours, and about 675 g of germinated differentiated seeds were collected.
4.12 g of the collected germinated differentiated seeds were put in a mortar, 10 ml of distilled water was added and crushed with a pestle. This crushed material was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract. 250 μl of this germinated seed extract was analyzed.
The relative value of the antioxidant power of the seed extract was measured by the absorbance A = log ₁₀ (I ₀ / I) using the diphenpicrylhydrazyl (DPPH) / Trolox radical scavenging activity method. Here, I ₀ is the light intensity in the absence of radicals, and I is the light intensity attenuated in the presence of radicals in the solution. This absorbance depends on the concentration of free radicals after the antioxidant in the extraction solution has captured. Table 3 shows the relative value of seed antioxidant power against DPPH radicals. By reducing the oxygen concentration below the ambient environment, the antioxidant power increases to at least 5 times more than 40 μmol DPPH equiv./gm-seed.

Example 7: Effect of carbon dioxide concentration The effect of carbon dioxide concentration on the germination induction process was examined using grape seeds. 225 g of grape seeds were washed with running water, and then immersed in a 100 ppm sodium hypochlorite solution at 40 ° C. for 4 hours. The seeds were then washed with distilled water and the water was removed with a soft paper. A gas in which seeds are put into a desiccator and the carbon dioxide concentration is adjusted to 2,000 ppm (0.2 vol%), 10,000 ppm (1 vol%), and 50,000 ppm (5 vol%) by mixing carbon dioxide gas and air. Was passed through a desiccator. Usually, the carbon dioxide concentration in the atmosphere is about 300 ppm. The oxygen concentration was 20 to 21% by volume under any conditions. The temperature was maintained at 28 ° C., and a 10 ppm sodium hypochlorite solution at 28 ° C. was sprinkled every 4 hours to induce germination for 48 hours, and about 675 g of germinated differentiated seeds were collected. 4.12 g of the collected germinated differentiated seeds were put in a mortar, 10 ml of distilled water was added and crushed with a pestle. This crushed material was centrifuged at 10,000 × g for 15 minutes, and the supernatant was used as an extract.
Table 4 shows the antioxidant activity value of the readily water-soluble polyphenol in the extract measured by diphenicpicrylhydrazyl (DPPH) radical scavenging activity. When the carbon dioxide concentration in the atmospheric air was exceeded, the antioxidant power became 40 μmol DPPH equiv./gm-seed, which is 8 times or more.

  The trend of increasing antioxidant capacity with increasing carbon dioxide concentration in the atmosphere is that many seeds (Asteraceae, Leguminosae, Cross Flower, Perillaceae, Calabidae, Rose, Ginkgo, Gramineae, Mulberry, Seeds of camellia, camellia, oleaceae, holly, specifically grapes, sunflower, safflower, clover, brassica, sesame, egoma, flaxseed, perilla, peanut, rice, buckwheat, corn, wheat, wild Rice, barley, millet, millet, fenugreek, rosemary, thyme, sage, mint, American red cherry, apricot, almond, grapefruit, orange, plum, St. John's wort, tomato, strawberry, carrot, bell pepper, mangosteen, Mango, loquat, burdock, cacao etc.).

Example 8: Effect of light irradiation In Examples 3 and 4, germination induction step at normal oxygen concentration and carbon dioxide concentration (21% oxygen concentration, 350 ppm carbon dioxide concentration) while irradiating different visible light doses And the analysis was repeated. The results are shown in Table 5. The light was irradiated using a fluorescent tube for 0 hour (dark condition), 2 hours, 6 hours, and 24 hours. The light intensity was 5000 lux with a simple illuminometer. Easily water-soluble polyphenols were extracted from seeds irradiated under each condition.
Compared to dark conditions, the highly water-soluble polyphenols extracted from seeds irradiated with light for at least 2 hours per day showed stronger antioxidant power. Even in the absence of light irradiation, the amount of antioxidant component increased with germination time, but it was found that the amount of antioxidant component in seed extract increased with light irradiation. Depending on the conditions, light irradiation may increase the antioxidant capacity of the seed by 50% or more.

This tendency is related to many seeds (Asteraceae, Leguminosae, Cross Flower, Perillaceae, Calcaretaceae, Rose, Ginkgo, Gramineae, Mulberry, Tadeidae, Camellia, Coleaceae, Holly, etc. In particular, grape, sunflower, safflower, clover, brassica, sesame, egoma, flaxseed, perilla, peanut, rice, buckwheat, corn, wheat, wild rice, barley, millet, millet, koroha, rosemary, thyme, Sage, Mint, American Red Cherry, Apricot, Almond, Grape Roots, Orange, Plum, St. John's Wort, Tomato, Strawberry, Carrot, Pepper, Mangosteen, Mango, Biwa, Burdock, Cacao, etc.).
Specific examples and embodiments of the means and apparatus are for the purpose of explanation, and various changes can be made without departing from the spirit and scope of the present invention. For example, the embodiments of the present invention can be applied to various systems, and easily water-soluble polyphenols and antioxidant compounds can be obtained from many types of plant seeds. Since the present invention can easily extract and purify polyphenols from seeds without using an organic solvent, it can be applied to a wide range of uses such as foods, health foods, pharmaceuticals, and cosmetics. Furthermore, in the embodiment, the extraction from the seed was performed using the aqueous solution, but the present invention can be applied to many other uses such as directly incorporating seeds or seed by-products into food as an example. It can be used for any application that utilizes water-soluble polyphenols. Furthermore, the characteristics of the embodiments may be incorporated in other embodiments, and those characteristics may not be described as one embodiment in the present description. Therefore, the above description should not limit the scope of the invention as defined in the appended claims.

The graph which shows the temperature change of 24 hours at the time of laminating | stacking grape seeds on the germination induction container 800 of FIG. The graph which shows the temperature change of 24 hours at the time of sprinkling a mung bean seed to the germination container 700 of FIG. The graph which shows the oxygen concentration change for 24 hours at the time of putting germinating seeds in the germination induction container 800 of FIG. FIG. 8 is a graph showing a change in oxygen concentration for 24 hours when mung bean seeds are placed in the germination induction container 700 of FIG. 7 and germination is induced at a room temperature of 28 ° C. or a room temperature of 36 ° C. FIG. FIG. 9 is a graph showing changes in carbon dioxide concentration over 24 hours when mungbean seeds are placed in the germination induction container 800 of FIG. 8 and germination is induced at room temperature of 28 ° C. FIG. FIG. 8 is a graph showing a change in carbon dioxide concentration for 24 hours when mung bean seeds are placed in the germination container 700 of FIG. 7 and germination is induced at a room temperature of 28 ° C. or a room temperature of 36 ° C. FIG. Explanatory drawing explaining the state in which the seed entered the germination container. Explanatory drawing explaining the state in which the seed entered the germination container.

Claims (25)

Increased water-soluble polyphenols that maintain germinable plant seeds under atmospheric conditions of carbon dioxide concentration of about 2000 ppm or more and / or oxygen concentration of about 18% by volume or less, and holding the plant seeds in the germination temperature range A method for producing germinated differentiated plant seeds. The method for producing a germinated differentiated plant seed according to claim 1, wherein the plant seed is hydrated. The method for producing germinated differentiated plant seeds according to claim 2, wherein the water content includes watering. The method for producing a germinated differentiated plant seed according to claim 1, comprising a step of adding water to the plant seed before the germination differentiation step. The method for producing germinated differentiated plant seeds according to claim 1, wherein the plant seeds are heated before the germination differentiation step. The method for producing germinated differentiated plant seeds according to claim 1, wherein the plant seeds are irradiated with light. The method for producing germinated differentiated plant seeds according to claim 6, wherein the irradiation with light is 5000 lux or more per day for 2 hours or more. The method for producing germinated differentiated plant seeds according to claim 1, wherein in the germination induction step, the carbon dioxide concentration in the atmospheric condition is 2,000 ppm or more and 50,000 ppm or less, and 60% or more of the germination induction step time is maintained. The method for producing germinated differentiated plant seeds according to claim 1, wherein, in the germination induction step, the oxygen concentration under atmospheric conditions is 4 vol% or more and 18 vol% or less, and 60% or more of the germination induction step time is maintained. The method for producing a germinated differentiated plant seed according to claim 1, wherein the germination temperature is 20 ° C. to 45 ° C. in the germination induction step. The method for producing germinated differentiated plant seeds according to claim 1, wherein the heating temperature is 5 ° C. or higher and 30 ° C. or lower, which is the median value of the appropriate germination temperature. The method according to claim 1, wherein the germination induction step is a germination induction step performed by depositing seeds, and temperature adjustment is performed using a temperature increase due to heat of germination and a temperature decrease due to watering. A readily water-soluble polyphenol produced by subjecting plant seeds to a germination induction step under atmospheric conditions where the carbon dioxide concentration is 2,000 ppm or more and / or the oxygen concentration is 18% by volume or less, and the temperature is maintained at 20 ° C to 45 ° C. And / or food containing antioxidants. The food according to claim 12, wherein the seed is subjected to moisture and / or light irradiation during the germination differentiation process. The food according to claim 14, wherein the irradiation with light is 5000 lux or more per day for 2 hours or more. The food according to claim 14, wherein the food is maintained at a temperature of 5 ° C. or higher and 30 ° C. or lower, which is the median optimal temperature for germination of plant seeds. The foodstuff of Claim 12 containing the easily water-soluble extract derived from a plant containing a polyphenol. The food according to claim 13, comprising plant seeds. The food according to claim 18, comprising plant seeds containing 1 milligram or more of a readily water-soluble polyphenol per gram of plant seeds. The food according to claim 18, comprising plant seeds containing 2 milligrams or more of a readily water-soluble polyphenol per gram of plant seeds. The food according to claim 18, comprising a plant seed containing an antioxidant equivalent to 40 µmol DPPH or more per gram of plant seed. The foodstuff of Claim 18 containing the plant seed containing 8 times or more easily water-soluble polyphenol compared with an ungerminated seed. The foodstuff according to claim 12 and claim 18, wherein the plant seed is basically composed of a grape seed. Water-soluble, having means for maintaining a carbon dioxide concentration of about 2,000 ppm or more and / or an oxygen concentration of about 18% by volume or less, maintaining the temperature of the plant seed within a suitable temperature range for germination, and allowing the plant seed to be hydrated An apparatus for germinating and differentiating germinated plant seeds with increased polyphenols. 25. The apparatus of claim 24, comprising a light source for irradiating plant seeds with light at 5000 lux or more for 2 hours or more per day.

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