Food and drink composition for green juice

Abstract

To provide a drinking composition for green juice which comprises cosmetic effects.SOLUTION: There is provided a drinking composition for green juice using a stem and / or leaf of barley, the barley satisfies following condition A, and satisfies one or more conditions selected from conditions (B)-(D): (A) the barley comprises a high level expression gene of serine protease Cxp1 in determination by a gene marker Te945; (B) the barley comprises a barley yellow mosaic virus resistance gene rym3 in determination by a gene marker k09554-AvaI; (C) the barley comprises an erysiphe pisi resistance gene ml-o in determination by a gene marker om2; (D) the barley comprises a barley yellow mosaic virus resistance gene rym5 in determination by a gene marker E31 / M41.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an edible or drinkable composition for green juice, which uses barley stems and / or leaves. [Background technology]

[0002] Barley is believed to be native to Central Asia and is an annual or biennial herb belonging to the Poaceae family. Barley stalks and / or leaves (hereinafter referred to as "stalks and leaves") are known as ingredients in health foods rich in vitamins, minerals, dietary fiber, amino acids, chlorophyll, SOD enzymes, etc., and are used in food and beverage compositions such as green juice, jelly, cookies, ginger drinks, yogurt, and supplements. Green juice is a product made from various processed products such as dried powder and squeezed juice containing green plant leaves, and is used as a health food that allows easy intake of vegetable ingredients (Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Application Laid-Open No. 2008-266147 Summary of the Invention [Problem to be solved by the invention]

[0004] Barley and its stems and leaves have been used for cosmetic purposes in the past, but there is a demand for barley stems and leaves with even greater cosmetic effects than before. [Means for solving the problem]

[0005] Therefore, the present inventors conducted extensive research into the composition of barley stalks and leaves to achieve greater cosmetic effects than previously possible, and discovered that the functionality of barley stalks and leaves differs depending on the barley genotype. Further research revealed that barley with a specific genotype determined by genetic markers has excellent collagenase inhibitory activity and can enhance cosmetic effects. Furthermore, the present inventors discovered that barley stalks and leaves with a specific genotype have high α-glucosidase inhibitory activity and are therefore effective for dieting.

[0006] The present invention is based on the above findings and provides an edible or drinkable composition for green juice using barley stems and / or leaves, The barley satisfies the following condition (A) and one or more conditions selected from (B) to (D): (A) The gene encoding the serine protease Cxp1 is highly expressed as determined by the gene marker Te945. (B) The strain possesses the yellow mosaic disease resistance gene rym3 as determined by the genetic marker k09554-AvaI. (C) The strain contains the powdery mildew resistance gene ml-o as determined by the genetic marker om2. (D) The strain possesses the yellow mosaic disease resistance gene rym5 as determined by the genetic markers E31 / M41. [Effects of the Invention]

[0007] According to the present invention, it is possible to provide a dietary composition for green juice that has excellent collagenase inhibitory activity and therefore has a high cosmetic effect. Furthermore, the dietary composition for green juice of the present invention also has high α-glucosidase inhibitory activity and is therefore effective in promoting dieting. [Brief explanation of the drawings]

[0008] [Figure 1] FIG. 1 shows the nucleotide sequence of the highly expressed Cxp1 gene in barley. [Figure 2] FIG. 2 shows the nucleotide sequences of primers for determining the type (low expression type or high expression type) of the Cxp1 gene. [Figure 3]FIG. 3 shows the nucleotide sequence of the region containing k09554-AvaI, a CAPS marker for determining the rym3 gene in barley. [Figure 4] FIG. 4 shows the base sequences of the primers for determining the rym3 gene. [Figure 5] Figure 5 shows the nucleotide sequence of the region containing the om2 marker in the upstream region of the ml-o gene in barley. [Figure 6] FIG. 6 shows primers for determining the ml-o gene. [Figure 7] FIG. 7 shows the nucleotide sequence of the eIF4E gene at the rym5 locus in barley. [Figure 8] FIG. 8 shows the base sequences of the primers for determining the rym5 gene. [Figure 9] FIG. 9 shows the evaluation results of collagenase inhibition rates for the Examples and Comparative Examples. [Figure 10] FIG. 10 shows the evaluation results of the α-glucosidase inhibition rate for the Examples and Comparative Examples. DETAILED DESCRIPTION OF THE INVENTION

[0009] Hereinafter, the composition for consumption or drinking for green juice of the present invention will be described based on preferred embodiments thereof.

[0010] One of the characteristics of the edible composition for green juice of this embodiment is the genetic makeup of the barley used.

[0011] Specifically, the barley used in this embodiment is characterized by satisfying the following condition (A) and one or more selected from conditions (B) to (D). Hereinafter, such barley will also be referred to simply as "specific barley." (A) As determined by the gene marker Te945, the specimen has a highly expressed gene for serine protease Cxp1 (hereinafter sometimes simply referred to as "Cxp1"). (B) The strain has the yellow mosaic disease resistance gene rym3 (hereinafter simply referred to as "rym3" or "gene rym3") as determined by the genetic marker k09554-AvaI. (C) It has the powdery mildew resistance gene ml-o (hereinafter sometimes simply referred to as "ml-o" or "gene ml-o") as determined by the genetic marker om2. (D) The strain has the yellow mosaic disease resistance gene rym5 (hereinafter sometimes simply referred to as "rym5" or "gene rym5") as determined by the genetic marker E31 / M41.

[0012] Serine protease Cxp1 is said to be involved in the degradation of malt proteins. Serine protease Cxp1 is also sometimes called serine carboxypeptidase Cxp1. In the present invention, it is simply referred to as Cxp1. High expression of Cxp1 is known to result in excessive degradation of malt proteins, adversely affecting growth.

[0013] rym5 and rym3 are believed to be involved in barley yellow mosaic disease, which causes yellow-green, elongated, smear-like spots and brown necrotic spots. rym5 is believed to be resistant to types I, II, IV, and V of the YMD virus, while rym3 is believed to be resistant to types I, II, III, and III of the YMD virus. YMD is known to cause serious damage to barley quality and yield, including reduced tillers, shorter plant height, and even death in severely affected plants.

[0014] ml-o is the powdery mildew resistance gene at the Mlo locus. Powdery mildew is caused by a type of mold called Erysiphe graminis. Infection causes white powdery lesions, which spread over the entire leaf as the disease progresses, rendering the plant unsuitable for use as an ingredient in green juice. Several loci are known for resistance to powdery mildew, including the Mla locus, Mlo locus, MJg locus, and MJk locus.

[0015] As described above, the barley genes described in conditions (A) to (D) are not known to have any relation to the cosmetic and diet effects of the dietary composition for green juice.

[0016] The barley used in this embodiment preferably contains (A) a highly expressed gene for serine protease Cxp1 and (B) the yellow mosaic disease resistance gene rym3, as this will more effectively achieve the beauty and diet effects. Furthermore, the barley preferably contains at least one of (C) the powdery mildew resistance gene ml-o and (D) the yellow mosaic disease resistance gene rym5, as this will more easily enhance the beauty effects, and it is even more preferable for the barley to contain both the (C) powdery mildew resistance gene ml-o and the (D) yellow mosaic disease resistance gene rym5. Most preferably, the barley satisfies all of the requirements (A) to (D).

[0017] The method for confirming the presence or absence of each gene will be described in detail below. <Highly expressed gene for serine protease Cxp1> In this embodiment, whether or not barley satisfies condition (A) is determined based on a CAPS (Cleaved Amplified Polymorphic Sequences) marker called Te945. Te945 is a genetic marker that utilizes a single nucleotide polymorphism (SNP) in the serine protease Cxp1 gene, and has been reported as an eQTL marker related to the level of expression of serine protease Cxp1 mRNA (Funct. Integr. Genomics, (2006) 6: pp. 25-35). The location of this single nucleotide polymorphism (SNP) is indicated by the double underlined portion in the serine protease Cxp1 gene sequence shown in Figure 1. The sequence in Figure 1 is a portion of the sequence disclosed in the DNA DataBank of Japan (DDBJ) under Accession No. CAJV010037927. The above-mentioned literature (Funct. Integr. Genomics, (2006) 6: pp. 25-35) states that the barley gene is "atgacat" and that the double underlined portion and its surrounding sequence in Figure 1 are "atgacat" and "atgacat" as shown in Figure 1. t Type II is "cttg" and Type II is "atgacat ccttg" and type I, and the expression level of CxpI is type I > type II. In this embodiment, the primers shown in FIG. 2 (F primer: SEQ ID NO: 2, R primer: SEQ ID NO: 3) are used as primers that sandwich this SNP position, and the region sandwiched between these primers is amplified by polymerase chain reaction (PCR), and the "cat c After treating with BseGI, a restriction enzyme that specifically cleaves the sequence "c", the DNA fragments after enzyme treatment are subjected to electrophoresis, and the base length of the DNA fragments is confirmed to determine whether or not they contain a high-expression gene for serine protease Cxp1 (type I). In the sequence of Figure 1, the positions corresponding to the primers in Figure 2 are single underlined. In the sequence shown in Figure 1, the fragment amplified by PCR using the primers in Figure 2 is 382 bp, and in the case of a low-expression type, a fragment of this base length is observed even after the above-mentioned restriction enzyme treatment. On the other hand, in the case of a high-expression type (cat c If the fragment has the sequence c), a band with a shorter base length (178 bp) will be detected upon cleavage with the above restriction enzyme. For a more definitive determination, when the above-mentioned band with a shorter base length is detected, it is preferable that another DNA fragment (band) be observed at the 202 bp position.

[0018] As described above, in this embodiment, determining whether a barley has a highly expressed Cxp1 gene does not require analyzing the entire nucleotide sequence of the Cxp1 gene by DNA sequence analysis. However, when such analysis is performed, it is preferable that the barley has the nucleotide sequence of the Cxp1 gene from positions 2176 to 2187 in the gene sequence disclosed in DDBJ under Accession No. CAJV010037927.

[0019] <rym3 yellow mosaic disease resistance gene> In this embodiment, whether barley satisfies the condition (B) is determined based on the CAPS marker k09554-AvaI linked to rym3. Specifically, when the amplification product amplified with the primers shown in Figure 4 is treated with the restriction enzyme AvaI, the DNA is cleaved by this enzyme treatment. do not have The gene sequence amplified by these primers is shown in Figure 3. The band length amplified using the primers in Figure 4 in the sequence of Figure 3 was 365 bp. be In Figure 3, the portions corresponding to the primers shown in Figure 4 are underlined. More specifically, PCR is performed using barley DNA as a template and the primers shown in Figure 8, the resulting amplified product is treated with the restriction enzyme AvaI, and the treated product is subjected to electrophoresis. If a fragment corresponding to 205 bp in length and a fragment corresponding to 156 bp in length are confirmed, the gene rym3 is detected. do not It shall be.

[0020] As described above, in this embodiment, determining whether or not a barley has the rym3 gene does not require analyzing the entire rym3 gene sequence by DNA sequence analysis. However, when such analysis is performed, it is preferable that the barley has the rym3 gene sequence from positions 148 to 512 in the gene sequence disclosed in DDBJ under Accession No. AV929051.1.

[0021] <Powdery mildew resistance gene ml-o> In this embodiment, whether or not condition (C) is satisfied is determined using om2, which has been reported as a genetic marker for detecting insertion / deletion mutations (In / del mutations) in the 5' upstream region (-2.3 kb) of the ml-o gene. In this embodiment, having the ml-o gene means having an insertion mutation detected by om2. An example of a barley gene sequence with this insertion mutation is shown in Figure 5. The gene sequence shown in Figure 5 is a portion of the gene with accession number Y14573.1 in DDBJ. In Figure 5, the double-underlined portion indicates the inserted sequence. Furthermore, primers proposed in Genome. Vol. 49, 2006, pp. 864-872 as primers used to detect insertion / deletion mutations using the genetic marker om2 are shown in Figure 6, and the sequences corresponding to these primers are single-underlined in Figure 5. In the example shown in Figure 5, the base length of the DNA fragment when a gene with an insertion mutation is amplified with the primers in Figure 6 is approximately 1000 bp, and the base length of the DNA fragment without an insertion mutation is approximately 750 bp. The presence or absence of ml-o can be determined based on the difference in base length of this DNA fragment. Specifically, the base length of the amplified product can be confirmed by electrophoresis of the amplified product amplified by PCR using barley DNA as a template and the primers shown in Figure 6. If a fragment of approximately 1000 bp is detected when the amplified product obtained by PCR amplification using the primers is subjected to electrophoresis, it is determined that the ml-o gene is present.

[0022] As described above, in this embodiment, the presence of the ml-o gene does not require analysis of the entire nucleotide sequence of the ml-o gene by DNA sequence analysis. However, when such analysis is performed, it is preferable that the barley has the sequence from positions 8106 to 9198 in the gene sequence disclosed in DDBJ under Accession No. Y14573.1 as the nucleotide sequence of the ml-o gene.

[0023] <rym5 gene for resistance to yellow mosaic disease> In this embodiment, whether or not barley satisfies condition (D) is determined based on E31 / M41, an STS marker derived from an AFLP polymorphism that has been reported to be linked to rym5 at 0.034 cM. A portion of the base sequence of the eIF4E gene at the rym5 locus is shown in Figure 7. The sequence shown in Figure 7 is listed in DDBJ under Accession No. AY661558. In this embodiment, PCR is performed using the primers shown in Figure 8 (reported in Theor. Appl. Genet. (2005) 110: pp. 283-293) derived from E31 / M41, and the PCR product is subjected to electrophoresis. The presence or absence of a DNA fragment band and the base length of the band are confirmed to determine whether or not the rym5 gene is present. In Figure 7, the portion corresponding to the primers shown in Figure 8 is underlined. In the sequence shown in Figure 7, the fragment amplified by PCR using the primers shown in Figure 8 is 436 bp. On the other hand, barley that does not have the rym5 gene usually does not produce an amplified product by PCR. When the amplified product obtained by PCR amplification using barley DNA as a template and the primers shown in Figure 8 is subjected to electrophoresis, if a fragment corresponding to a length of 436 bp is detected, the barley is deemed to have the yellow mosaic disease resistance gene rym5.

[0024] As described above, in this embodiment, the determination of the rym5 gene does not require analysis of the entire nucleotide sequence of the rym5 gene by DNA sequence analysis. However, when DNA sequence analysis is performed on the barley used in this embodiment, it is preferable that the barley used in this embodiment has the sequence from positions 44169 to 44604 in the gene sequence disclosed in DDBJ under Accession No. AY661558 as the nucleotide sequence of the eIF4E gene at the rym5 locus.

[0025] When determining the presence or absence of the genes described in (A) to (D) using the above genetic markers, the barley DNA to be determined can be extracted from the flowers, seeds, fruits, leaves, stems, and roots, and it is preferable to use barley seeds.

[0026] Specific test methods include, for example, the following methods. <Gene presence / absence determination test> (1) DNA is extracted from barley. This extraction can be carried out using a commercially available kit or the like. The resulting DNA solution is preferably adjusted to a DNA concentration of 1 ng / μL to 20 ng / μL before being subjected to PCR. (2) PCR is carried out using the DNA solution obtained in (1) as a template. PCR is preferably carried out under the conditions shown in Table 1 below. Primers that can be used include those shown in Figures 2, 4, 6, and 8 above.

[0027] [Table 1]

[0028] (3) When determining the presence or absence of the highly expressed Cxp1 gene and rym3 in the PCR product obtained in (2), enzyme treatment is performed using the restriction enzymes listed in Table 1. The restriction enzyme treatment is preferably performed at 30°C to 40°C for 30 minutes to 24 hours. The restriction enzymes used were FastDigest BseGI (Thermo Fisher) for BseGI and 2,000 units of AvaI (New England Biolabs) for AvaI. The resulting treated product is used as a sample for electrophoresis, which can be carried out by any known method.

[0029] The specific barley may be botanically classified as six-row barley, two-row barley, or naked barley, and may be any of a wild species, a hybrid, etc.

[0030] Furthermore, in this embodiment, whether or not conditions (A) to (D) are satisfied may differ from the actual traits of the barley. For example, barley determined to have the (A) Cxp1 high-expression gene based on the above genetic markers may not necessarily have the trait of high expression of serine protease Cxp1. Similarly, barley determined to have (B) rym3 or (D) rym5 based on the above genetic markers may not necessarily have the trait of yellow mosaic disease resistance. Barley determined to have (C) ml-o based on the above genetic markers may not necessarily have the trait of powdery mildew resistance. Furthermore, when a barley is determined to have satisfied any of conditions (A) to (D) corresponding to the marker in a determination using any of the above genetic markers, the barley is still considered to have the gene, even if a different determination result would be obtained using a different method.

[0031] The stalks and leaves of the above-mentioned specific barley are preferably harvested before maturity, i.e., from the time when tillers begin to emerge until the time when heading begins. Specifically, although this differs depending on the variety, it is generally preferable to harvest stalks and leaves from barley that is 10 cm or taller, particularly about 10 to 90 cm, and especially about 30 to 60 cm, but this is not limited thereto. Barley stalks and leaves are preferably processed immediately after harvest. If time is required before processing, they are stored by a storage method commonly used by those skilled in the art, such as low-temperature storage, to prevent deterioration of the barley stalks and leaves.

[0032] In the dietary composition for green juice of this embodiment, various processed products obtained from the specific barley stems and leaves can be used as the stems and leaves. Examples of such processed products include crushed stems and leaves and their dried powder (crushed powder), shredded stems and leaves and their dried powder (shredded powder), squeezed stem and leaf juice and its dried powder (squeezed juice powder), and stem and leaf extract and its dried powder (extract powder).

[0033] For example, conventionally known methods can be used to pulverize barley stems and leaves. One such method is a method in which the barley stems and leaves are subjected to a drying treatment and a pulverization treatment in combination. Either the drying treatment or the pulverization treatment may be performed first, although it is preferable to perform the drying treatment first. This pulverization method may be further combined with one or more treatments selected from blanching, sterilization, and the like, as necessary. The pulverization treatment may be performed once, or two or more treatments may be combined, although it is preferable to perform a coarse pulverization treatment followed by a fine pulverization treatment to pulverize the barley stems and leaves more finely.

[0034] Blanching is a treatment for maintaining the vivid green color of stems and leaves, and examples of blanching methods include hot water treatment and steam treatment. The blanching treatment is preferably performed by treating the stems and leaves in hot water or steam at 80 to 100°C, preferably 90 to 100°C, for 60 to 180 seconds, preferably 90 to 120 seconds. When hot water treatment is used as the blanching treatment, dissolving a carbonate such as magnesium carbonate or a bicarbonate such as sodium bicarbonate in the hot water is preferred, as this can make the green color of the stems and leaves more vivid. The steam treatment is preferably an intermittent steam treatment, in which the stems and leaves are repeatedly steamed with steam and cooled under normal or increased pressure. In the intermittent steam treatment, the steam treatment is preferably performed for 20 to 40 seconds, more preferably 30 seconds. Cooling treatment after steaming is preferably carried out immediately. The method is not particularly limited, but includes immersion in cold water, refrigeration, cooling with cold air, evaporative cooling with hot air, and evaporative cooling using a combination of hot and cold air. Of these, evaporative cooling using a combination of hot and cold air is preferred. Such cooling treatment is carried out so that the product temperature of the stems and leaves is preferably 60°C or less, more preferably 50°C or less, and most preferably 40°C or less. In addition, to produce stem and leaf powder rich in nutrients such as vitamins, minerals, and chlorophyll, it is preferable to repeat the intermittent steaming treatment 2 to 5 times.

[0035] Sterilization is a process that usually uses temperature, pressure, electromagnetic waves, chemicals, etc. to physically or chemically kill microbial cells. When blanching is performed in addition to drying and grinding, it is preferable that the blanching be performed before the drying. When sterilization is performed in addition to drying and grinding, it is preferable that the sterilization be performed after the drying, or before or after the grinding.

[0036] The drying treatment is preferably a treatment to dry the stems and leaves to a moisture content of 10% or less, particularly 5% or less. This drying treatment can be carried out by any method known to those skilled in the art, such as hot air drying, high-pressure steam drying, electromagnetic wave drying, freeze-drying, etc. Drying by heating can be carried out at a temperature of preferably 40°C to 140°C, more preferably 80°C to 130°C, for a time period that does not cause discoloration of the stems and leaves.

[0037] The pulverization process can be performed by any method commonly used by those skilled in the art, such as using a crusher, mill, blender, or stone mill. The pulverized stems and leaves are sieved as needed, and those that pass through a 30 to 250 mesh sieve are preferably used as stem and leaf powder. By using particles with a particle size of 250 mesh or smaller, the stem and leaf powder can be easily handled during further processing, and by using particles with a particle size of 30 mesh or larger, the stem and leaf powder can be easily mixed uniformly with other materials.

[0038] Specific examples of pulverization methods include cutting barley stems and leaves, blanching them, drying them to a moisture content of 10% by mass or less, preferably 5% by mass or less, and then pulverizing them (see JP 2004-000210 A). Other examples include cutting barley stems and leaves, blanching them, rolling them, drying them, and pulverizing them (see JP 2002-065204 A and Japanese Patent No. 3428956 A). Other examples include drying barley stems and leaves, coarsely pulverizing them, heating them at 110°C or higher, and then finely pulverizing them (see JP 2003-033151 A and Japanese Patent No. 3277181 A).

[0039] Barley stems and leaves can be shredded by methods commonly used by those skilled in the art when shredding plants, such as slicing, crushing, or chopping. One example of shredding is slurrying. Slurrying is performed by subjecting barley stems and leaves to a mixer, juicer, blender, mass colloider, or the like, to turn the barley stems and leaves into a mushy gruel (a suspension of liquid and solids). By slurrying the barley stems and leaves in this manner, the stems and leaves are shredded so that 80% by mass or more of the shredded pieces have an average diameter of preferably 1 mm or less, more preferably 0.5 mm or less, even more preferably 0.1 mm or less, and most preferably 0.05 mm or less, and the shredded material becomes fluid. The resulting shredded material can also be freeze-dried, hot-air dried, or otherwise processed to produce a dry powder (shredded powder).

[0040] Methods for extracting juice from barley stems and leaves include compressing barley stems and leaves or shredded barley stems and leaves, or centrifuging or filtering shredded barley stems and leaves. A typical example involves extracting juice using a mechanical crushing device such as a mixer or juicer, and then removing coarse solids by sieving, filtration, or other means, to obtain a juice. Specific examples include those described in JP-A-08-245408, JP-A-09-047252, JP-A-5-7471, and JP-A-4-341153, among others. Those skilled in the art can select and implement these known methods as appropriate. The obtained juice may be concentrated as needed, or may be subjected to processes such as freeze-drying, hot air drying, or spray drying to produce a dried powder (juice powder). When producing a juice powder, the juice alone may be dried, or it may be dried together with an excipient as needed.

[0041] One method for obtaining a barley stem and leaf extract involves adding an extraction solvent commonly used by those skilled in the art, such as ethanol, water, or aqueous ethanol, to barley stem and leaf or shredded barley stem and leaf material, and stirring or heating as necessary to extract. The resulting extract may be concentrated as needed, or may be freeze-dried, hot-air dried, spray-dried, or otherwise processed to produce a dried powder (extract powder). When producing an extract powder, the extract alone may be dried, or it may be dried together with an excipient as needed.

[0042] In the present invention, of the processed products of barley stems and leaves, it is preferable to use dried powder (pulverized powder, shredded powder, squeezed powder, extract powder), with pulverized powder or squeezed powder being more preferable, and pulverized powder obtained by drying and crushing the stems and leaves is particularly preferable because it can make the edible composition for green juice have a brighter color and a better flavor, can be made rich in dietary fiber, and has a high cosmetic effect. Pulverized powder is produced without going through a juicing process.

[0043] The edible composition for green juice of the present invention may contain only barley stems and leaves, or may contain other ingredients as described below. The content of barley stems and leaves in the solid content of the edible composition for green juice of the present invention is preferably 0.1% by mass or more, more preferably 0.5% by mass or more, even more preferably 1.0% by mass or more, particularly preferably 10.0% by mass or more, and most preferably 20.0% by mass or more, on a dry mass basis. The upper limit is preferably 99.9% by mass or less, more preferably 90.0% by mass or less, and particularly preferably 80.0% by mass or less.

[0044] Furthermore, the proportion of the stems and leaves of the specific barley in the barley stems and leaves contained in the edible composition for green juice is preferably 10.0% by mass or more, more preferably 30.0% by mass or more, particularly preferably 50.0% by mass or more, and most preferably 70.0% by mass or more.

[0045] There are no particular restrictions on the amount of intake of the food or beverage composition of the present invention, but from the perspective of more significantly exerting the effects of the present invention, it is preferable for an adult to ingest 10 mg or more of the stems and / or leaves of the barley of the present invention per day, more preferably 50 mg or more, and even more preferably 100 mg or more.

[0046] The edible composition for green juice may contain other ingredients in addition to the six-row barley stems and leaves. Examples of such other ingredients include vitamins, proteins, oligosaccharides, minerals, polysaccharides, dairy products, processed plant products, microorganisms such as lactic acid bacteria, sugars, sweeteners, citric acid, acidulants, coloring agents, thickeners, and glazing agents, as well as manufacturing agents such as talc, silicon dioxide, cellulose, and calcium stearate. Other ingredients include various excipients, binders, lubricants, stabilizers, diluents, bulking agents, thickeners, emulsifiers, coloring agents, flavorings, food additives, and seasonings. The content of such other ingredients can be appropriately selected depending on the form of the food or beverage. In this embodiment, it is particularly preferable that the edible composition for green juice contains oligosaccharides, water-soluble dietary fiber, and lactic acid bacteria as ingredients other than the processed barley stems and leaves.

[0047] The composition for food and drink for green juice of this embodiment can be in any form, for example, a form suitable for oral ingestion such as eating or drinking, specifically, powder, granules, tablets, rods, plates, blocks, solids, rounds, liquids, candies, pastes, creams, capsules such as hard capsules and soft capsules, caplets, tablets, gels, jelly, gummies, wafers, biscuits, cookies, cakes, chewable tablets, syrups, sticks, etc.

[0048] The edible and beverage compositions for green juice of this embodiment include green juice, juices, shakes, smoothies made by adding fruit juice, vegetables, dairy products, etc. to green juice, and soft drinks, carbonated drinks, and the bases for these drinks.

[0049] The edible composition for green juice is preferably in a powder form (powder, granules, or other powder form) and in a form in which the mixture is mixed with water and taken orally, as this prevents spoilage, makes it suitable for long-term storage, and the edible composition has a vivid color when mixed with water. Furthermore, if the edible composition for green juice is in a solid form, as described above, it can be mixed with water to form a liquid and then taken orally, for example, by drinking the liquid. However, depending on the preferences of the consumer, the solid form may also be taken orally. Furthermore, the edible composition may be added to not only water, but also milk, soy milk, fruit juice drinks, whey drinks, soft drinks, yogurt, pancake mix, etc. Needless to say, it may also be used as a food with nutritional function claims, a food for specified health uses, or a food with functional claims.

[0050] As described above, the dietary composition for green juice of this embodiment has collagenase inhibitory activity because it uses barley stems and leaves that satisfy the above conditions, as determined by the above genetic markers. Collagenase is an enzyme that degrades collagen. Collagen, like hyaluronic acid and elastin, is found in mammalian connective tissue and is a component that contributes to skin moisture retention, lubrication, and flexibility. Collagen activity increases with inflammation and aging, leading to increased collagen degradation. As a result, the balance between collagen synthesis and degradation is disrupted, leading to problems such as increased inflammatory responses and aging-related skin wrinkles and joint pain. Therefore, ingredients that inhibit collagenase enzyme activity are expected to prevent inflammation, wrinkles, joint pain, and other issues associated with collagen reduction. Furthermore, the dietary composition for green juice of this embodiment uses barley stems and leaves that satisfy the above-mentioned conditions, thereby exhibiting excellent α-glucosidase inhibitory activity. α-Glucosidase is a digestive enzyme that digests glucose, starch, and other substances. Inhibiting this enzyme can suppress the absorption of carbohydrates and promote their excretion from the body. As a result, excess energy intake into the body is suppressed, fat accumulation in the body is inhibited, and accumulated fat can be reduced, resulting in excellent diet effects. As used herein, "diet effect" refers to the effect of preventing, preventing, or reducing obesity, slimming down, or reducing body weight in mammals such as humans. The diet effect referred to herein also includes effects such as body fat reduction, suppression of body fat accumulation, visceral fat reduction, and subcutaneous fat reduction, which are due to α-glucosidase inhibitory activity and result in the prevention, prevention, or reduction of obesity, slimming down, or weight loss. Therefore, the edible composition for green juice of this embodiment has excellent α-glucosidase inhibitory activity, and is expected to have the effect of suppressing the digestion and absorption of sugar, suppressing blood sugar level increases, preventing diabetes and diabetes complications, and providing a diet effect. Therefore, the edible composition for green juice of this embodiment can be used for one or more purposes selected from the following: inhibiting collagen-degrading enzymes, suppressing collagen reduction, maintaining collagen, reducing joint pain and inflammation, preventing skin wrinkles, improving skin lubricity, flexibility and water retention, cosmetic purposes, suppressing sugar digestion and absorption, suppressing blood sugar level increases, preventing diabetes or diabetic complications, and dieting.

[0051] Specifically, the food and beverage compositions of the present invention can be used as pharmaceuticals (including quasi-drugs) or so-called health foods, and examples of such uses include products labeled with claims such as "maintains skin moisture," "provides a skin barrier," "protects the skin," "for those concerned with blemishes," "for those concerned with wrinkles," "skin that can withstand stress," "good for beauty," "good for the skin," "for knee and hip problems," "for a healthy gait," "for lighter movements," "regulates joint function," "smooths joint movement," "improves joint function," "relieves joint movement problems," "helps joint movement," "supports joint movement," "for those concerned about blood sugar levels," "moderates rises in blood sugar levels," "moderates sugar absorption," "suppresses blood sugar levels," "for those concerned about body fat," "for those slightly overweight," "for those concerned about weight (BMI)," "reduces weight and abdominal fat (visceral fat and subcutaneous fat)," "reduce waist circumference," "for those wishing to achieve their ideal body shape," and "for those wishing to achieve a slim body shape." [Example]

[0052] The present invention will be explained in more detail below by showing examples, but the scope of the present invention is not limited to these examples.

[0053] <Examples 1 to 3, Comparative Examples 1 to 3> The above-mentioned "gene presence / absence determination test" was carried out on seeds of multiple varieties. That is, DNA extraction, PCR, restriction enzyme treatment as required, and electrophoresis were carried out using the above-mentioned method using the gene markers shown in Table 1. The specific conditions and procedures were as follows. DNA extraction: Several seeds were collected from several barley plants of different varieties. DNA was extracted from some of the seeds derived from one plant. DNA was extracted using the Dneasy PlantMini Kit (QIAGEN). The extraction method followed the protocol provided with the kit. PCR: The DNA obtained above was diluted with nuclease-free water to a DNA concentration of 7.5 ng / μL and used in PCR. PCR was performed using the DNA solution after adjusting the concentration as a template and mixing the following reagents in the amounts shown in Table 2 below, and the resulting solution was subjected to the conditions shown in Table 1 above. In Table 2, Ex Taq and its buffer are products of Takara Bio Inc.

[0054] [Table 2]

[0055] Restriction enzyme treatment: The resulting PCR product was diluted 2-fold with 20 μl of nuclease-free water. To determine the presence or absence of (A) a highly expressed Cxp1 gene and (B) the yellow mosaic disease resistance gene rym3, the restriction enzymes in Table 1 above were used, and the solutions in Table 3 below were mixed in a tube in the proportions shown in Table 3 below. The sealed tube was then submerged in a 37°C water bath, incubated for 1 hour, and then cooled on ice. In Table 3 below, the restriction enzyme buffer used was the buffer provided with the enzyme. The resulting treated product was used as the sample for electrophoresis. To determine other genes, the PCR product was diluted 2-fold with nuclease-free water as described above without restriction enzyme treatment and used as the sample for electrophoresis.

[0056] [Table 3]

[0057] Electrophoresis: 0.5x TBE (Tris Borate EDTA) buffer and agarose were added to a flask so that the agarose concentration was 1% or 2% by mass, and the mixture was dispersed. The flask was autoclaved to dissolve the agarose, and then gently stirred. Once cooled, the mixture was diluted with GelGreen (BIOTIUM). A small amount of 0.5x TBE buffer was added and the mixture was stirred again. An agarose gel (hereafter referred to as AGE) was formed using a special mold. 0.5x TBE buffer and AGE were placed in an electrophoresis chamber (Mupid α Mini Gel Electrophoresis Chamber; Advance). 0.5x TBE buffer was added until the gel was completely submerged. 1 μL of 6x Loading Buffer (Takara Bio Inc.) and 5 μL of sample were mixed and then applied to each well of the AGE. Electrophoresis was performed at 100 V.

[0058] Seeds obtained from the same plants as those for which the following results were obtained using the above method were designated as barley for the Examples and Comparative Examples according to the correspondence in Table 4. In Table 4 below, ○ indicates that the result was that the corresponding gene was present, and × indicates that the result was that the corresponding gene was not present.

[0059] [Table 4]

[0060] The barley seeds from each of the Examples and Comparative Examples for which the above judgments were obtained were sown and cultivated using known methods, and the stems and leaves were collected before maturity, i.e., from the time when tillering begins to occur until the time when heading begins. The pulverized powder was obtained using the method described below, and this was used as a powdered composition for consumption or drinking of green juice.

[0061] <Method of manufacturing dry powder> Harvested barley stems and leaves were washed with water to remove any adhering mud, and then pre-treated by cutting into pieces approximately 5 cm in size. The pre-treated stems and leaves were blanched once in hot water at 90-95°C for approximately 100 seconds, and then cooled in cold water. The resulting stems and leaves were then dried in a dryer with hot air at 70-80°C until the moisture content was 5% by mass or less. The dried stems and leaves were coarsely crushed to a size of approximately 1 mm using a mixer, and then finely crushed using a jet mill grinder to obtain crushed barley stem and leaf powder.

[0062] The obtained compositions for eating and drinking for green juice of Examples 1 to 3 and Comparative Examples 1 to 3 were subjected to the following evaluations.

[0063] <Rating 1: Beauty effect> (1) After autoclaving the powdered dietary composition for green juice, the concentration was adjusted to 4 mg / mL with phosphate-buffered saline (PBS), and a sample solution was prepared at a concentration four times the final concentration. 50 μL of this sample solution was added to each well of a 96-well black plate. However, 50 μL of PBS was added to some wells as a control instead of the sample solution. (2) 100 μL / well of the enzyme solution (Test) or 0.1% BSA-containing D-PBS(-) (Blank) was added to a 96-well black plate and incubated at 37°C for 10 minutes. D-PBS(-) was prepared by dissolving one tablet of Wako Pure Chemical Industries' PBS tablet (Model No. T900) in 100 mL of purified water. Albumin (bovine serum-derived Cohn Fraction V, Wako Pure Chemical Industries) was added to this solution to a concentration of 0.1% by mass to produce 0.1% BSA-containing D-PBS(-). The enzyme solution was prepared by diluting a 10 mg / mL solution of collagenase B (Roche) in D-PBS(-) with 0.1% BSA-containing D-PBS(-) to a concentration of 10 μg / mL. (3) After incubation, 50 μL / well of fluorescent substrate solution (a 1 mM DMSO solution of MOCAc-Pro-Leu-Gly-Leu-A2pr(DNP)-Ala-NH2 (Peptide Institute) adjusted to 5 μM in D-PBS(-) containing 0.1% by mass of BSA) was added to the 96-well black plate, and the plate was incubated at 37°C for 60 minutes in the dark. (4) Using a microplate reader, the fluorescence intensity was measured at 405 nm with excitation at 320 nm. (5) Using the measured fluorescence intensity, the collagenase inhibition rate was calculated using the following formula: The results are shown in Figure 10. Collagenase inhibition rate (%) = [1 - (Sample test - Sample blank ) / (Control test - Control blank )] x 100 In the above formula, Sample testis the fluorescence intensity of the well to which the sample solution and enzyme solution were added, and Sample blank indicates the fluorescence intensity of the well to which the sample solution was added and no enzyme solution was added (0.1% BSA-containing D-PBS(-) was added), and Control test indicates the fluorescence intensity of the well to which the enzyme solution was added (PBS was added) and no sample solution was added. blank is the fluorescence intensity of the well to which neither the sample solution nor the enzyme solution was added.

[0064] From FIG. 9, it can be seen that the dietary and beverage composition for green juice of the present invention, which has a specific genotype determined by a specific gene marker, has excellent collagenase inhibitory activity and is effective for beauty.

[0065] <Rating 2: Diet effect> The α-glucosidase inhibitory activity of the dietary and beverage compositions for green juice of Examples 1 to 3 and Comparative Examples 1 and 2 was evaluated by the following method.

[0066] Experimental Method Sample solutions were prepared by dispensing 80 μL of each of the dietary and beverage compositions for green juice prepared in Examples 1 to 3 and Comparative Examples 1 and 2. Next, 40 μL of a 0.02 M aqueous substrate solution (substrate: p-nitrophenyl-α-D-glucopyranoside; Wako Pure Chemical Industries, Ltd.) was added to each sample solution, and the mixture was incubated at 37° C. for 5 minutes.

[0067] Thereafter, 40 μL of phosphate buffer containing 2 mg / mL BSA and 0.3 μg / mL α-glucosidase (Wako Pure Chemical Industries, Ltd.) (enzyme solution) was added, and the mixture was further maintained at 37° C. for 15 minutes.

[0068] Next, 160 μL of 0.2 M aqueous sodium carbonate solution was added as a stop solution, and the resulting solution was used as a test solution and its absorbance at 440 nm was measured (measured value A). As a control, the absorbance of a control test solution obtained in the same manner as above except that purified water was used instead of the sample solution was measured (measured value B).

[0069] Furthermore, as blanks for the test solution and the control test solution, the absorbance of each solution (blank) obtained using the same procedure as above, except that the stop solution was added before the enzyme solution, was measured (the measurement value for the blank of the test solution was designated as measurement C, and the measurement value for the blank of the control test solution was designated as measurement D).

[0070] Using the above measured values ​​A to D, the α-glucosidase inhibition rate (%) was calculated using the following formula (n=3): α-Glucosidase inhibition rate (%) = (1-[AC] / [BD]) x 100 The results are shown in Figure 10. As shown in Figure 10, it was found that the edible and beverage compositions for green juice of Examples 1 to 3 have superior α-glucosidase inhibitory activity compared to Comparative Examples 1 and 2. Therefore, the edible and beverage compositions for green juice of Examples 1 to 3 have an excellent diet effect.

[0071] [Production of the composition of the present invention] [Production Example 1] (Production of granules for instant drinks) The following ingredients were mixed to produce 3g of granules for instant drinks. Dissolving the obtained granules for instant drinks in 100mL of water and taking it will provide excellent beauty effects.

[0072] 40% by mass of ground barley stalks and leaves that meet the conditions (A) and (D) but not the conditions (B) and (C) Indigestible dextrin 5% by mass Erythritol 5% by mass Fragrance 2% by mass Dextrin Remainder [Production Example 2] (Production of granules for instant drinks) The following ingredients were mixed to produce 4g of granules for instant drinks. Dissolving the obtained granules for instant drinks in 100mL of water and taking it will provide excellent beauty effects.

[0073] 60% by mass of barley stalk and leaf juice powder that satisfies the conditions (A), (C), and (D) but not the condition (B) Crushed kale powder 10% by mass Matcha 3% by mass Trehalose 20% by mass Vitamin B 0.2% by mass Vitamin C 0.2% by mass Dextrin Remainder

Claims

[Claim 1] A composition for consumption or drinking for green juice containing crushed barley stems and / or leaves, The barley satisfies all of the following conditions (A), (B), (C), and (D), and the dietary composition for green juice is characterized in that it is for dieting. (A) The strain has a highly expressed gene for the serine protease Cxp1 as determined by the gene marker Te945. (B) The strain has the yellow mosaic disease resistance gene rym3 as determined by the gene marker k09554-AvaI. (C) Carries the powdery mildew resistance gene ml-o as determined by the gene marker om2. (D) Contains the yellow mosaic disease resistance gene rym5 as determined by the genetic markers E31 / M41.

Images