JP6646356B2 - Method for producing plant powder containing rare sugar - Google Patents

Description

The present invention relates to a method for producing a plant powder containing a rare sugar derived from a plant, and in particular, Zyna or Ryub (Zyuna is called Ryob in some regions. In the present invention, Ryub-Zyna. omit this description.) powder manufacture of, and makes it possible to be utilized as health foods.

  2. Description of the Related Art Conventionally, a technique of drying a plant such as a leaf or a stem to form a powdered tea or food material, or using a medicinal plant for a powdered health food or the like has often been performed. For example, for the purpose of obtaining Moroheiya dry powder or Moroheya tea with less oxalic acid, after cutting the Moroheya leaves and stems to appropriate dimensions, steaming with live steam for several minutes, then drying while rubbing, and then crushing (Patent Literature 1), a method in which green leaves of a grass plant having a high concentration of GABA having a blood pressure lowering action are microwave-treated, and then dried to obtain a dry powder (Patent Document 2). Are known. In addition, a method characterized by a drying step for producing a dried green powder in which the dried powder of a green plant is retained without losing the color and active ingredients of green leaves has been proposed (Patent Document 3).

In the drying by heating mainly with hot air, the dried powder changes color to brown or the like, and the appearance is poor, and the components are often denatured due to heat, denatured, reduced in content, or extinguished. Discoloration and alteration can be reduced by additives or the like, but the freeze-drying method is often employed because it is not easy to produce a powder composed of the colors and components inherent in the plant.
For example, a succulent plant is sterilized and washed using sodium hypochlorite in a solid state, and when vacuum freeze-dried, the powder production process of drying at low temperature and for a long time allows the succulent plant to have only the inherent colors and components. (Patent Literature 4), a plant belonging to the genus Umbelliferae or the genus Kawaraboufu, a kind of eucalyptus such as a eucalyptus, an inu cane or a hyuga tou uki, or a plant or so-called mountain ginseng A method for producing a raw material that can minimize component loss and deterioration by freeze-drying sun-dried leaves, stems, and seeds into powder (Patent Document 5), a state in which okra is freeze-dried as it is By pulverizing into a powder state, it can be used as a food additive in various dishes, and pulverized into a powder state with the okra frozen and dried as it is And that step, the steps of a tablet form by adding an excipient and a thickener such as starch to the okra powder, by producing a supplement. Furthermore, a method of producing okra tea by a step of pulverizing okra in a state of being freeze-dried in a state of being lyophilized into a powder state and a step of mixing the okra powder with a substance such as barley or hub tea (Patent Document It is known to produce a dry powder of a test substance by a freeze-drying method such as 6).
The use of edible Apios (Apios americana Medicus) flowers as a food material that suppresses an increase in blood sugar level has been proposed (Patent Document 7). This is due to β-D-glucopyranosyl (E) -3- (3,4-dihydroxyphenyl) acrylate (caffeoyl β-D-glucopyranoside) isolated from Apios flowers, or 2-O- [3- ( 3,4-dihydroxyphenyl) acryloyl] -D-glucopyranose was found to suppress an increase in blood sugar level in normal mice and diabetic model mice, and uses Apios flowers as a health food material.

JP 2003-18975 A JP-A-2002-58446 JP 2007-252340 A JP 2009-248991 A JP 2008-303174 A JP 2008-303174 A JP 2014-87364 A International Publication No. 2010/113785

Abstracts of Annual Meeting of the Biotechnology Society of Japan 2007, 108, 2007-08-02 Phytochemistry 48 (2): 241-248. Biosci. Biotechnol. Biochem. 2006, 70 (9), 2081-2085. Life Sci. 2007, 81 (7), 592-599. Journal of Japan Society of Nutrition and Food Vol.59 (2006) No.2 P119-121 Japan Society for Food Science and Technology Vol.57 (2010) No.6 P263-267

With the spread of preventive medicine knowledge, attempts have been made to maintain health by ingesting naturally derived foods having bioactivity prophylactically in a healthy or semi-healthy state prior to illness, and using natural materials Has been put on the market as a health food. Among them, there are many that claim an anti-diabetic effect, but there are few cases where the effect has been proven. As far as the present inventors know, rare sugars are harmless natural products that can be added to the original food, do not affect taste, and foods and drinks having safe natural products themselves have not yet been developed. . That is, no report has yet been made to verify that plants themselves that do not correspond to extracts from natural products have characteristic physiological activities that improve one or more of blood sugar levels, weight gain, and visceral fat accumulation. It is believed that there is a strong social need for safe, natural products with excellent characteristic bioactivity that improve one or more of blood sugar levels, weight gain, and visceral fat accumulation.
The present invention provides that a plant body such as a leaf of a chin or a ryobu plant contains a rare saccharide having a physiological activity, and the rare saccharide improves one or more of blood sugar level, weight gain, and visceral fat accumulation. Based on being a characteristic physiologically active substance, it aims to promote health promotion by providing supplements such as health foods and supplements for these plants.

An object of the present invention is to provide a characteristic bioactive substance which improves the stability of blood sugar, weight gain, and one or more of visceral fat accumulation with high stability and low side effects consisting of a natural plant itself. I do.
It is another object of the present invention to find a growing method in which the content of rare sugars is increased in growing natural plants, and as a result, to provide a plant powder having an increased content of rare sugars.

It is known that plants such as leaves and stems of Zinnia or Ryobu contain rare sugars having physiological activity. In providing foods and beverages containing rare sugars and health foods and beverages, we realized dry powdered Zyuna or Ryobu so that plants containing the rare sugars can be used in various forms. We were able to. Further, in consideration of the fact that the dry powder concentrates the rare sugar content, a treatment method for increasing the rare sugar content of the plant was found.

The gist of the present invention is a method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to the following (1) to ( 11 ).
(1) A dormancy-breaking Zuina or Ryobu seed is obtained through a process in which the harvested Zuina or Ryobu seed is subjected to a dormancy breaking treatment by storing it in a dry place at 4 ° C. in a dark place for a period of 2 to 6 months. The bred seeds or Ryub seeds are germinated in a light place at a temperature of 25 to 30 ° C., and the germinated seeds are transplanted to a culture medium or a culture soil to obtain a Zyna or Ryobu plant, and the stem from the young plant of the Zyna or Ryobu. A step of cutting a plant or a plant containing a stem apex or a stem from a seedling in which the 2-5th leaf of the plant of Zyna or Ryobu is expanded, or the cut plant or plant of Ryub or Ryub as it is. Or simply remove the leaves and place them on the rooting medium or pierce them like cuttings and cultivate them. To obtain a rooted and rooted Zuina or Ryobu plant, or to obtain an adult bud, leaf or branch, or a germinated seed from the plant, and produce a plant grown by passaging or tissue culture. And a method for producing a plant powder containing the rare sugar D-psicose derived from a plant, wherein the powder is produced by subjecting any of the plants to drying and grinding.
(2) The method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to the above (1), wherein the plant powder is a supplement-shaped processed food.
(3) The above-mentioned (1) or (1), wherein the plant powder is a functional food having a function based on the rare sugar D-psicose that improves one or more biological functions of blood sugar level, weight gain, and visceral fat accumulation. 2. A method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to 2).
(4) The improvement in blood sugar level of the plant powder based on the rare sugar D-psicose is necessary for saving rare insulin which is a physiological activity of the rare sugar, improving insulin sensitivity, and correcting a hyperglycemia. The method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to the above (3), which is effective in improving symptoms or preventing the onset of disease.
(5) The plant-derived rare sugar D-psicose according to any one of (1) to (4), wherein the rare sugar D-psicose is contained in a plant powder containing the rare sugar D-psicose in an amount of 0.1 g or more. A method for producing a plant powder comprising:
(6) The method for producing a plant powder comprising the plant-derived rare sugar D-psicose according to any one of the above (1) to (5), wherein the component other than the rare sugar D-psicose is a plant-derived powder.
(7) The method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to the above ( 6 ), wherein the component other than the rare sugar D-psicose is 40 to 95% by weight of the whole.
(8) The plant powder containing the rare sugar D-psicose derived from a plant according to the above (6) or (7), wherein the component other than the rare sugar includes a powder of a plant body of Chinese radish or Ryobu. Production method.
(9) The production of a plant powder containing the rare sugar D-psicose derived from a plant according to any one of the above (1) to (8), wherein the drying and pulverizing is freeze-drying and pulverizing. Method.
(10) a plant powder containing a rare sugar D- psicose plant-derived, to the content of rare sugars D- psicose above (1) to have increased by the addition of new rare sugar D- psicose (9) A method for producing a plant powder comprising the rare sugar D-psicose derived from a plant according to any one of the above.
(11) The plant-derived rare sugar D- according to any one of the above (1) to ( 10 ), to which a step of adding a powder that becomes a drink by drinking water from Japanese tea, black tea, Chinese tea, or water is added. A method for producing a plant powder containing psicose.

Plants, such as leaves and stems of Zuina or Ryobu, contain rare sugars having physiological activity, and the rare sugars improve one or more of blood sugar level, weight gain, and visceral fat accumulation. Based on being an active substance, health promotion can be promoted by providing supplements such as health foods and supplements of these plants.
It is possible to provide a characteristic physiologically active substance which is composed of a plant which is a natural product itself, has high stability, improves blood glucose level, weight gain and one or more of visceral fat accumulation with few side effects.

It is known that plants such as leaves and stems of Zinnia or Ryobu contain rare sugars having physiological activity. Rare sugars have characteristic biological activities that improve one or more of blood sugar levels, weight gain, and visceral fat accumulation, such as diabetes, latent diabetic conditions, obesity, hyperlipidemia, arteriosclerosis It is useful for prevention and treatment. In providing foods and drinks and health foods and drinks containing rare sugars, to realize a zinna or ryobu consisting of a dry powder so that plants containing the rare sugars can be used in various forms. I was able to.
Natural drying, drying with a desiccant, freeze-drying, etc. are known as processing methods for the purpose of long-term storage and ease of use of the plant. Therefore, it is difficult to maintain the color, aroma, and contained components. Therefore, the present invention produces a dry powder while maintaining the physiological activity of rare sugars contained in the plants of Chinese radish and Ryobu, and utilizes this. Thus, it can be used as a health food or a drug having a physiological activity.
Moreover, since it is made of plant powder collected from the natural world, it can be made a health food that is gentle to the human body and gentle to the human body. Furthermore, the finding of a treatment method for increasing the rare sugar content of a plant is significant in view of the fact that the dry powder concentrates the rare sugar content.

The relative value of the change of the average length in 10 days in each treatment section of Chinese pine is shown. A list of the substances used for the treatment of Zinnia and the list of treatments are shown below. The substances used for the treatment of Zinnia and the treatment summary (details) are shown below. The result list (plant hormone, sugar) of Sisukaba is shown. The list of Siskaba results (heat treatment, scratch treatment, salt) is shown.

[Plants containing rare sugars]
The plant containing the rare sugar D-psicose is only the genus Zinnia (Non-Patent Document 1). It is also known to inhibit the growth of many plants. An insulin secretion effect (Non-patent document 2), an effect of preventing arteriosclerosis (Non-patent document 3) and the like are known. It has about 70% the sweetness of sugar, but has almost no calories. In animal experiments with rats, “slow increase in blood glucose level after meal” (Non-patent document 4), “suppression of visceral fat accumulation” (Non-patent document 5), and “hard to atherosclerosis” (Non-patent document 6) ) Has been reported. However, it has been reported that the effect of suppressing the increase in blood glucose level after eating is affected by the decrease in psicose due to heating, and the effect is reduced as compared with the non-heated state. In the case of cooking at a high temperature such as Karinto, the effect is more greatly reduced. The effect of inhibiting the growth of nematodes has been confirmed, and it may be used as an anthelmintic. The present invention relates to a plant powder containing a rare saccharide produced by freeze-drying and crushing a plant of Zinnia or Ryobu, and to easily utilize the rare saccharide contained in these plants. Further, the present invention is to obtain a powder which can be used as a food material or a material for a health promotion agent by adopting a form which can be widely applied.
As described above, rare sugars have the properties of assimilating sugars, excellent sweeteners, antiobesity agents, food depressants, insulin sensitizers, and low-calorie sweeteners. It has new characteristics, such as causing food suppression. In addition, since the sweetness is close to that of sugar and the calorie is low, it is attracting attention as a sugar that can be widely used as a low-calorie sweetener.
Rare saccharides are monosaccharides excluding D-glucose, D-mannose, and allitol, which are abundant in nature. For example, D-psicose and D-allose are monosaccharides that are produced in very small amounts in nature. However, its properties and uses are still being explored. At present, rare sugars such as D-psicose can be produced by using enzymes and the like, but it has been found that rare sugars are also contained in plants such as leaves of japonica and ryobu.
For example, it is known that the leaves of D. japonica contain D-psicose at about 5% of the fresh weight among the rare sugars. In the present invention, for the purpose of utilizing the rare sugars contained in these plant objects, the rare sugars are contained by pulverizing a plant body including leaves, buds, stems, roots and the like of Chinese radish. It is used as powder. For example, harvesting of a plant from Zuina or Ryobu may be performed by harvesting leaves from an adult tree, or by transplanting whole plants including leaves, stems and roots of seedlings germinated from seeds, tissue-cultured or subcultured seedlings. A powder can be obtained by collecting and freeze-drying.
In the present invention, commercially available freeze-drying equipment is used for freeze-drying the Chinese plant. Since the freeze-drying method is a lower-temperature treatment than other drying methods, it does not destroy nutrients and is excellent in restorability. Preventing tissue destruction due to dehydration is also a feature of this drying method. Since food is nearly 90% water, great care must be taken against spoilage. Lyophilization is used in the production of preserved foods because the final degree of drying is much better than other drying methods. In addition, the weight of the dried food is reduced by the amount of water removed, so that the food is suitable for transportation. A powder of a plant of Zinna or Ryobu is produced from a plant of Zinina or Ryobu without impairing the functional components of the plant of Rosa or Rina as much as possible. For example, a step of pulverizing a plant of Zinnia or Ryobu to make a slurry, a step of pre-freezing the slurry in a divided freezing mold, and a vacuum freeze-drying process of reducing the frozen product to a water content of 3 to 5% are performed. And a step of adjusting the powder of the dried product obtained from the step. In addition, vacuum drying and spray drying are applied as a drying process.

In the following, the cultivation and freeze-drying of Zuina as an example will be described in detail.
As plant values containing rare sugars, there are presently known plant values, such as "china" and "ryobu".
[Zuina]
Zuina is a plant belonging to the genus Zuina. The genus Zina (Zea genus, genus Rina, scientific name: Itea japonica ) is a deciduous shrub of the family Saxifragaceae and is a genus of plants consisting of about 10 shrubs. The young leaves are boiled and eaten like yomena, so they are also called yomenonaki, the trunk is 1-2 meters in height, the shoots are pale green, the leaves are alternate, the oval oblong shape is sharp, the tip is sharp and the edges are fine There are saw teeth. About 8 pairs of parallel side veins stand out on the leaf surface. From May to June, the branches have an upward inflorescence of 5 to 17 cm in length, and many white florets open. The petals are egg-shaped lanceolate, five erect, with the same number of petals on both the gaku and stamens, one pistil, head stigma, and ovary in the upper half. Zuina grows in the warm mountains and is distributed in the southern Kinki region, Shikoku and Kyushu. Zinnia is known to be the only plant that produces D-psicose and allitol contained in rare sugars in plants. It is also sold as a seedling in Japan.

[Ryobu]
Ryobu is a plant belonging to the family Ryobu, and is described in the botanical picture book as follows.
Distributed on Jeju Island in Hokkaido, Honshu, Shikoku, Kyushu and the Korean Peninsula. It is a small deciduous tree that grows in the forest. It is only 3 to 7 meters high, its trunk is smooth and brown, and its branches are ring-shaped. The shoot has star-shaped hair. The leaves are stalked and alternating, broadly covered needle-shaped with toothed teeth at the edges, gathered at the tip of the branches, sparsely hairless or star-shaped hairs, and dense hairs on the back vein. In summer, shoots of inflorescences with a length of 6 to 15 cm appear at the tips of the branches, and small white flowers are densely attached. The sepal is small and splits 5 times. The corolla is 5-6 mm in diameter, is deeply cleaved, and resembles a small plum blossom. 10 stamens and 1 pistil. A spherical, small fruit. The wood becomes high quality charcoal, and the sprouts are edible. Ryobu does not contain the rare sugar psicose, but in some regions Zuina is sometimes called Ryobu. The present invention is for Zuina.

[Cultivation from Zuina seed]
Since the seeds of Zuina are dormant, they do not germinate by sowing unless the dormancy is broken. In order to break the dormancy of seeds, it is necessary to store them in a cool, dark place in a dry atmosphere for a predetermined period of time. This is done by storing for a period. It is preferable that the water vapor pressure is low as the drying conditions. For example, it is stored together with a large amount of silica gel in a sealed plastic container. The storage temperature is 4 ° C. or lower, preferably 4 ° C. Further, the storage period under such conditions is preferably 2 weeks or more, preferably 2 to 6 months. Since the seeds of the Chinese radish are light germination seeds, germination is difficult when covered with soil, and germination is preferably performed at a temperature of 25 to 30 ° C. in a light place. The germination rate after two weeks at 28 ° C. is about 60%. It is.
In order to grow germinated seeds, for example, germinated seeds are grown in the same 1/2 MS medium [containing 1% sucrose (pH 5.8)] or are planted in soil and grown in the same manner as ordinary plants. Can be. When the germinated seeds are planted in a medium or soil, the growth speed is not inferior to that of the seeds, but the cultivation of the soil is slightly more lignified. The transplantation of the germinated seeds can be performed, for example, about 10 days after sowing.

[Growth by tissue culture]
The seedlings in which the 2-5th leaf of the Chinese radish developed, or the Chinese vegetative plant containing the shoot apex or the stalk of a normal Chinese radish seedling, or the leaves were removed, and were directly used in a 1/2 MS medium (1% sucrose) There is a method of cultivating a Chinese radish which enables a large amount of seedlings to be proliferated by merely laying it in a plant culture medium (with or without addition) or by piercing it like a cutting. As a Chinese plant, a seedling obtained by breaking dormancy of a seed improved by crossing and making a seedling can be used. Germination of seeds that have broken dormancy and mass production of young seedlings by subculture thereof make it possible to easily obtain large amounts of plants or seedlings of Chinese quince.

[Growth of Chinese yam seedling by subculture]
In order to mass-produce the Chinese seedlings, a part of the plant can be taken out and cultured aseptically in a test tube or the like by tissue culture. For example, a large amount of seedlings can be easily produced by culturing cells near the growth point at the tip of the stem. For example, a plant (approximately 5-10 mm) containing the shoot apex of a young seedling in which the 2-5th leaf has been developed is cut out, pierced into a 1/2 MS medium like a cutting, or cultured by merely placing the stem. . A plurality of side buds were observed from the lower part of the plant (plant including the root and cotyledon or first leaf) whose stem apex was cut. Since lateral shoots are formed by cutting the shoot apex, the growth point can be easily obtained. In the case of stems without leaves alone, even if they are placed on the medium, homogeneous young leaves from all are observed.

A medium in which 3 μM, 5 μM, and 10 μM of indolebutyric acid (IBA) were added to a 1/2 MS medium, and a medium without addition were prepared, and cuttings were performed on the medium. In approximately 10 days, any medium with IBA3-10 μM was developed. Roots were induced. However, since the roots were formed earliest in the non-supplemented medium and the roots grew well, it is considered that plant hormones (such as IBA) are not particularly necessary.
Regarding the subculture, either a cutting at the shoot apex or a cutting at the shoot (not including the shoot apex) may be used. Rooting is generally performed within 10 days after transplantation into a 1/2 MS medium (containing 1% sucrose). I do. In the case of a stem without leaves alone, homogeneous young leaves from all of them and rooting of some of them are observed after 30 days just by placing in a medium.

[Zuina seedling making]
Plants subjected to rooting treatment (cultivated until rooting for one month or more) in a petri dish are transplanted to culture soil (Sakata seeds: supermix), and preferably covered with a food such as a transparent film. After 2-3 days, the food is shifted to remove moisture, and after 2-4 days, the food is completely removed. Regarding this acclimatization step, if a container with a hood is used, there is no problem even with a seedling of only futaba. Further, it is preferable to prevent insect damage such as larvae of Spodoptera litura by covering insect repellent nets during the seedling raising period. The rooted Chinese plant is transplanted to a plug tray medium or culture soil and grown in the form of a plug seedling.
If the outside temperature decreases during growth (the minimum temperature falls below 15 degrees), the leaves turn red. Zuina does not fall, but Koba no Zuina leaves in winter. Since the growth of the seedlings is greatly affected by the temperature, it is better to start the acclimatization step from early spring and grow them for summer, and in winter it is more efficient to concentrate on the above subculture work and to grow seedlings from early spring Good schedule to work on.

[Growth of Zuina seedling]
In the same manner as in the production of a seedling of a juvenile plant, a juvenile plant containing a stem portion from a young tree of the plant or a plant plant containing a stem portion or a stem portion from a seedling in which the 2-5th leaf of the plant was developed was cut and cut. The roots of the Chinese plants are placed in a rooting medium, and they are cultivated for rooting. Vegetables and / or new leafy vegetables are grown until they can be harvested, and the young leafy and / or new leafy vegetables are harvested. As the rooting medium, a 1/2 MS medium or a Gamburg B5 medium is used. When growing a rooted Chinese vegetative plant into a Chinese vegetative vegetable, the use of a medium makes it possible to aseptically grow a functional vegetable based on the functions of D-psicose and allitol. In addition, Example 4 shows a method of growing a Chinese radish (a pretreatment method) for increasing the rare sugar content.

[Rare sugar]
Rare sugars are defined as monosaccharides and sugar alcohols that are present in trace amounts in nature. As for hexose (hexose), in the case of aldose, L-allose, L-gulose, L-glucose, L-galactose, L-altrose, L-idose, L-mannose, L-talose, D-talose, 16 kinds of D-mannose, D-idose, D-altrose, D-galactose, D-glucose, D-gulose, D-allose, and in the case of ketose, L-psicose, L-sorbose, L-fructose, L- There are eight types: tagatose, D-tagatose, D-fructose, D-sorbose, and D-psicose. For example, there is no report that D-psicose or D-allose has toxicity to humans, and it is considered that toxicity to animals is low.
As a method for obtaining D-psicose from a rare sugar, at present, a production method obtained by treating fructose with an enzyme (epimerase) is generally used. As a method for obtaining D-allose, a method of synthesizing from D-psicose using L-rhamnose isomerase and a method of obtaining D-allose by allowing D-xylose isomerase to act on a solution containing D-psicose are known. Moreover, it can be obtained by a technique disclosed in Patent Document 8 using rare sugar syrup and isomerized sugar as raw materials. Manufactured mainly to contain D-psicose and D-allose, but also 0.5 to 17% by weight of D-psicose, 0.2 to 10% by weight of D-allose and other unidentified rare sugars It is. The rare sugar syrup (trade name: Rare Sugar Sweet) thus produced contains about 15% (about 6% of D-psicose) of rare sugars such as glucose and fructose as main components such as D-allose and D-psicose. .

[Zuina freeze-dried]
(1) The roots of the subcultured subcultured (48 days) were removed with forceps and the agar was removed as much as possible.
(2) The raw weight of Chinese yam was 1 to 1.5 g per petri dish, and 50 g of fresh weight was collected from 35 petri dishes.
(3) 50 g of Zuina was divided into about 4 equal parts, wrapped in aluminum foil, immersed in liquid nitrogen and frozen.
(4) It was confirmed that the soybean was completely frozen by soaking it for 3 minutes or more.
(5) The frozen Chinese quince was divided into two plastic beakers (500 ml), covered with parafilm, and several holes were made.
(6) The Zuina put in the beaker was put in FREEZE DRY SYSTEM / FREEZONE 2.5 (LABCONCO) and freeze-dried for 48 hours.
(7) After drying, the weight was 5.88 g, which was about 1/10.
(8) The weighed Zuina was transferred to a mini speed mill.
(9) The mini speed mill was crushed for 10 seconds, but the grains were still coarse.
(10) When it took more 10 seconds, the grains were still a little coarse.
(11) When it was further taken for 10 seconds, the grains became moderately fine, and there was no significant change even if it took more time.
(12) The crushed ducks were collected as much as possible using a spoon or a spatula and placed in a 50 ml centrifuge tube.
(13) The weight of the ground powder collected was 5.44 g. The lid was wrapped with parafilm, protected from light with aluminum foil, placed in a box, and stored at room temperature in the dark.
As described above, Zuina could be lyophilized to a powder. After 24 hours of lyophilization, water remained in the zinnia, but after 48 hours almost no water remained. The weight of the dry matter of the final product was about 1/10 of the raw weight of the original Chinese yam. Even if it was made into a powder and stored at room temperature in the dark, it was found that the zinna was in a powder form and could be stored in this state. As an improvement, it was wrapped in aluminum foil by the procedure of freezing using liquid nitrogen, but this method requires a long time when put in FREEZE DRY. It is preferable to use a method of directly freezing Chinese varieties and freeze-drying them.

Ingredients: Sprout of Kovanozina Medium: 1/2 MS agar medium [containing 1% sucrose (pH 5.8), Agar 0.7%], agar is used as a support, but there is no particular designation.
<Surface sterilization>
Since the plants in the open field were collected, the sterilization conditions for the aerial parts were relatively long.
Cut off the leaves of the young shoots with a scalpel (be careful not to damage the side buds) and wash gently with household detergent.
After cutting the young shoots of Kobanozuina to an appropriate size, the shoot is treated with 70% ethanol for 2-3 minutes.
Twenty-20 drops of 2 drops are added to 100 ml of sodium hypochlorite solution (effective chlorine concentration 0.5%), and ethanol-treated juvenile shoots (about 4-7 cm) are immersed for 25-30 minutes. Transfer the sterilized watermelon young shoots to sterilized Milli-Q water, replace the sterilized water with new one every 20 minutes (four times), cut the obtained surface sterilized watermelon young shoots into appropriate size (about 2 cm), Transfer to new medium.
It became possible to obtain homogeneous young leaves for Kobanozuina.
The cultivated forty-eight days were extracted with tweezers, and a young plant, consisting of young leaves, stems and roots, was collected. 50 g of this plant was collected, stored in an aluminum container, immersed in liquid nitrogen for 3 minutes or more, and confirmed to be completely frozen. Next, it was stored in a plastic container and freeze-dried for 48 hours using FREEZE DRY SYSTEM / FREEZONE 2.5 (LABCONCO). After drying, the weight of the plant was 5.88 g, which was about 1/10. The dried plant body after the weight measurement was crushed at 25,000 rpm for about 30 seconds using a mini speed mill (MS-09, Shinion Co., Ltd.). The recovered powder weighed 5.44 g. When the ground sample was a leaf, it was finely ground (about 50 μm), and when it was a root / stem, it became a slightly larger powder (about 150 μm).
It has been found by analysis that rare plant sugars such as D-psicose and allitol are contained in the Zinnia plant in an amount of 1%, and the powder contains approximately 10% of the rare sugar in calculation. become. In addition, although the test sample of the Chinese quince used what was cultivated by light irradiation, when it was left dark for a certain period, the D-psicose content increased.
It was found that the properties of the powder and the rare sugar contained therein did not change even when the powdered Chinese flour was stored in a dark room at room temperature and under refrigeration (4 ° C.) for a long period of time.

  Fifty grams of leaves were collected from adult Chinese radish trees, poured into liquid nitrogen, rapidly frozen, and dried using a freeze dryer. Preliminary freezing temperature: about -40 ° C, shelf heating temperature during drying: 30 ° C, drying time: about 48 hours, and a dried product having a weight of 5.6 g after drying was obtained. The dried product was further ground with a mortar and pestle to obtain leaf tissue. The powder of the Chinese plant contains about 0.5 g of rare sugars such as D-psicose and allitol.

  A rare sugar syrup (liquid sugar containing about 10 to 15% of a rare sugar such as D-psicose) in 2 g of the dried powder of Zinna obtained in Example 1; trade name “Rare Sugar Sweet” (released by Rea Corporation) Sweet, seller: Matsutani Chemical Industry Co., Ltd.) was added and mixed so as to be 50 parts by weight with respect to 100 parts by weight of the dry powder, and dried at 80 ° C. to increase the rare sugar content. 3 g of the dried powdered Chinese radix contains about 0.3 g of rare sugar.

[Purpose]
Trying to treat various varieties of Zuina to see if the psicose content increases. If the conditions under which psicose can be increased can be clarified, the value of Zuina will increase greatly, and at the same time, the contribution to research in new Zuina will be significant. The growth promotion effect of gibberellin was also confirmed. [Preparation]
Zuina (2-4 weeks after passage)
Reagents of various concentrations in a 24-well plate (eg: 100 mg / l GA ₃ (dissolve in a small amount of ethanol, then make up with Milli Q water)
Tweezers Parafilm Kim Wipe [Method]
(1) Reagents (eg, in the case of gibberellic acid (GA ₃ ), 100 μg / l, 1 μg / l, 10 μg / l1, 50 μg / l, 100 mg / l) are sprinkled to each concentration (pH = 5.7 ± 0.1). ), 2 ml was put into a well of a 24-well plate. Milli-Q water was used as a control, and 1 ml of milli-Q water was put between each well. With the lid closed, a parafilm was wound and placed in a 28 ° C. chamber overnight.
(2) The Zina was allowed to stand for 15 minutes with the lid of the culture vessel tapper open before the treatment. After that, the cucumber was carefully pulled out of the medium with tweezers, and the medium attached to the root was removed by washing with water.
(3) After removing the culture medium attached to the roots by washing with water, the water content was removed with a Kimwipe, and the whole length was photographed.
(4) After the measurement, add zinna to each well.
(5) Place the plate in a tapper filled with water, cover the plate, and incubate at 28 ° C.
(6) Change to a new plate every two days, and measure the total length.
(7) Measure until the 10th day, weigh on the 11th day, and freeze and store.
[result]
FIG. 1 shows the relative value of the change in the average length over 10 days in each treatment section. The average length of each individual on day 0 is set to 1, and the average length of the relative value for each number of days is shown. Since the growth promoting effect of gibberellin is observed, it seems that the substance can be treated by this method. This time, elongation measurement was performed to confirm that the effect of gibberellin appeared, but it is not always necessary.
Psicose measurement etc. are shown below.

[Growth effect of gibberellin]
As shown in FIG. 1, the promotion effect of gibberellin was recognized in Chinese bean. This result indicates that this process is effective for obtaining a large number of Chinese varieties.

[Changes in psicose production due to various treatments]
Then, under the same conditions, various pretreatments were performed to examine changes in the amount of psicose in Chinese pine.
[Preparation]
Zuina Instant Dry Yeast (Japan-France Trading Co.)
2.0ml screw cap micro tube (sarstat)
Stainless steel beads (Tomy SUB-50)
50 mM phosphate buffer pH 7.5 (200 mM NaH ₂ PO ₄ and 200 mM Na ₂ HPO ₄ are mixed 4: 1 and diluted 4 times)
[Yeast solution preparation]
Weigh 1 g of instant dry yeast, suspend in 10 ml of Buffer Buffer, and centrifuge at 4000 rpm for 2 min. The supernatant is removed, and the suspension resuspended in 10 ml of Buffer Buffer is used as a yeast solution.
[Sample preparation]
(1) The weight of the sample 11 days after the substance treatment frozen in liquid nitrogen is measured and recorded.
(2) A frozen twiner is placed upside down in a 2.0 ml screw cap microtube (sarstat) containing two stainless steel beads (Tomy SUB-50), and the whole tube is frozen with liquid nitrogen.
(3) Crush at 3000 rpm for 1 min using Micro Smash MS-100 (Tomy).
(4) Add 500 μl of water, sufficiently suspend, and centrifuge at 4 ° C. for 10 minutes at full speed.
(5) Transfer the supernatant to a 1.5 ml tube, and transfer 250 μl of the supernatant to a 1.5 ml tube containing 250 μl of yeast solution.
(6) The tube containing the yeast solution is permeated at 30 ° C. and 180 rpm for 30 minutes.
(7) After centrifugation for 1 min, the supernatant was recovered, and 20 μl of the supernatant was used as a yeast-treated sample.

[Cysteine carbazole sulfate method]
The cysteine carbazole sulfate method (ciskaba) is an improved method of the Kalha & Sour & sulfuric acid method. Dische & E. The procedure reported by Borenfreund (1951) gives a purple-red color, showing strong specificity for keto sugars. This method is referred to as "cis cover" for convenience.
[reagent]
1.70% sulfuric acid Cysteine reagent (1.5% Cysteine-HCl aqueous solution, adjust at the time of use)
3. Carbazole reagent (0.12% Carbazole 100% alcohol solution, adjust at the time of use)
[operation]
One cycle (24-well plate) required amount (1) Dissolve 10 mg of Milli-Q in 150 mg of L-Cysteine hydrochloride monohydrate (2) Dissolve 12 μg of Carbazole in 10 ml of 100% ethanol.
(3) Add 6 ml of Milli-Q to 14 ml of H ₂ SO ₄ .
(4) Add 20 μl of sample to each well of 1.5 ml.
(5) Add 20 μl Cysteine solution to each well.
(6) Let react for 1 minute at room temperature.
(7) Add 600 μl 70% H ₂ SO ₄ to each well.
(8) Add 20 μl Carbazole solution to each well.
(9) Let stand at room temperature for 20 minutes → The solution discolors.
(10) Measure absorbance (540 nm) with an absorbance meter.
[Summary of Siskaba results]
FIG. 2 shows a list of the substances used for the processing of the Chinese pine and the processing. FIG. 3 shows the substances used for the processing of the Chinese pine and the processing summary (details). FIG. 4 shows a list of the results (plant hormones, sugars) of the cispava. FIG. 5 shows a list of the results of heat treatment (heat treatment, scratch treatment, salt). However, the ordinates in FIGS. 3 and 4 show relative (%) and Errorbar = Standard Error when Control (water) is set to 100.

[Summary of the amount of psicose in Chinese cabbage by various pretreatments]
As shown in FIG. 4 and FIG. 5, it has been clarified that the amount of psicose in the varieties varies greatly by performing various processes. Some are reduced by pre-processing, while others are increased. In particular, it is very useful to be able to increase the psicose in Chinese pine by performing the pretreatment.
Increasing the content of psicose will greatly increase the commercial value of the plant. In addition to creating value as a new product, clarifying this mechanism has given great implications for the study of metabolism in Zuina.

The present invention relates to a dry powder of a plant in order to utilize a rare sugar having bioactivity, which is contained in a plant such as a leaf or a stem of Zinnia or Ryobu. The physiological activities of rare sugars include poor insulin conservation, improved insulin sensitivity, and diseases that require correction of hyperglycemia, and improvement of symptoms or suppression of abnormal daily rise in plasma glucose concentration (blood glucose). Since it has a preventive effect, it is possible to provide foods and health foods and drinks which are useful for prevention and treatment of diabetes, potential diabetic conditions, obesity, hyperlipidemia, arteriosclerosis and the like. By making a plant containing a rare sugar into a dry powder, a dry powder that can be used in various forms could be obtained. It is very useful to be able to increase the psicose in the Chinese pine by performing the pre-processing.

Claims (11)

  The harvested Zuina or Ryobu seeds are subjected to a dormancy breaking treatment by storing them for 2 to 6 months in a dry place at 4 ° C. in a dark place to obtain dormant breaking Zuina or Ryobu seeds. Ryobu seeds are germinated in a light place at a temperature of 25 to 30 ° C., and the germinated seeds are transplanted to a culture medium or a culture soil to obtain a Zuina or Ryobu plant, and a Zuina containing a stem from a young tree of the Zuina or Ryobu. Or a step of cutting a plant or a plant containing a stem apex or a stem from a young seedling in which the second to fifth leaves of the leaf of R. japonica or Ryobu are developed. Just remove it and place it on the rooting medium or pierce it like a cutting. Rooting and rooting to obtain a plant of root or root, or obtaining a bud, leaf or branch of an adult tree or a germinated seed from the plant, producing a plant grown by passaging or tissue culture, and A method for producing a plant powder containing a rare sugar D-psicose derived from a plant, wherein the powder is produced by subjecting any one of the plants to drying and grinding.   The method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to claim 1, wherein the plant powder is a processed food in a supplement form.   The plant according to claim 1, wherein the plant powder is a functional food having a function based on the rare sugar D-psicose that improves one or more biological functions of blood sugar, weight gain, and visceral fat accumulation. A method for producing a plant powder containing the rare sugar D-psicose derived from the plant.   The plant powder improves the blood sugar level based on the rare sugar D-psicose, conserves poor insulin which is a physiological activity of the rare sugar, improves insulin sensitivity, and ameliorates the symptoms of diseases that require correction of hyperglycemia. Alternatively, the method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to claim 3, which is effective in preventing disease onset.   5. The plant powder containing the rare sugar D-psicose derived from a plant according to any one of claims 1 to 4, wherein the rare sugar D-psicose is contained in the plant powder containing the rare sugar D-psicose in an amount of 0.1 g or more. Production method.   The method for producing a plant powder containing the plant-derived rare sugar D-psicose according to any one of claims 1 to 5, wherein the component other than the rare sugar D-psicose is a plant-derived powder. The method for producing a plant powder containing the plant-derived rare sugar D-psicose according to claim 6 , wherein the component other than the rare sugar D-psicose comprises 40 to 95% by weight of the whole.   The method for producing a plant powder containing the plant-derived rare sugar D-psicose according to claim 6 or 7, wherein the component other than the rare sugar includes a powder of a plant body of Chinese radish or Ryubu.   The method for producing a plant powder containing the rare sugar D-psicose derived from a plant according to any one of claims 1 to 8, wherein the drying and pulverizing are freeze-drying and pulverizing. Plant powder containing a rare sugar D- psicose plant-derived, new according to any one to the content of the rare sugar D- psicose claims 1 has increased 9 by adding rare sugar D- psicose A method for producing a plant powder containing the rare sugar D-psicose derived from a plant. 11. A plant powder containing the rare sugar D-psicose derived from a plant according to any one of claims 1 to 10 , wherein a step of adding a powder that becomes a drink by water supply of Japanese tea, black tea, Chinese tea, or water is added. Production method.