KR101161231B1 - Isoflavone contents increasement by radiation in soybean - Google Patents

Abstract

The present invention relates to a method for increasing the isoflavone content of soybean seeds, and more particularly, to a certain time at the time when the seeds are enlarged in a pod of one of the top 4 nodes which have developed a fully developed leaf during soybean growth. It is a method of increasing the isoflavone content of seeds by treating the light for a certain period of time so that the beans are continuously exposed to light.

Bean seeds, isoflavone content, lighting

Description

Isoflavone contents increasement by radiation in soybean}

The present invention relates to a method for increasing the isoflavone content of soybean seeds.

Soybeans are a very important crop as a major source of protein and fat in our diet, which uses cereals as a staple food and vegetables as a side dish in our agricultural life. Soybean seeds are the main constituent of soybean seeds, which generally contain 40% protein, 20% fat, 35% carbohydrate, and 5% other ingredients. In Korea, various soy foods such as tofu, miso, soy sauce, and cheongukjang are consumed as a source of vegetable protein and fat. In addition, soybeans contain secondary metabolites that have many physiological activities. Especially, isoflavones are phytoestrogens that are found in soybeans, which help cancer, heart disease, osteoporosis and menopausal diseases in women. Known to [Tham et al. 1998; Jacobsen et al. 1998; Messina 2000; Adlercreutz 2002]. Isoflavones compete with estrogens, bind to estrogen receptors, and act as competitive inhibitors or antagonists for cancer treatment.Its cholesterol-lowering effects, cardiovascular disease, menopausal symptoms, menopausal symptoms and osteoporosis prevention, aging, inflammation, It is reported that the effect is excellent in diabetes. So far, isoflavones present in soybean have three types of glucosides (daidzin, glycitin and genistin) based on daidein, glycidin and genistein, and three kinds of aglycone. ), 3 types of malonyl glucosides (6 ^〃 - O -malonyl-7- O -daidzin, 6 ^〃 - O -malonyl-7- O -glycotin and 6 ^〃 - O -malonyl-7- O -genistin) and 3 Acetyl glucosides (6 ^〃 - O -acetyl-7- O -daidzin, 6 ^〃 - O -acetyl-7- O -glycotin and 6 ^〃 - O -acetyl-7- O -genistin). et al. 2004; Kim et al. 2005; Rostagno et al. 2004]. Among these substances, recent reports have shown that aglycone forms have higher absorption and bioavailability [Hutchins et al. 1995], the method of quantifying the total amount after converting to the aglycone form, which is used in the body has become popular. Although there have been many studies to quantify the differences between isoflavones and varieties, there are few studies on the genetic background. However, there have been reports of differences in isoflavone contents due to environmental variations and differences between growing regions.

Particularly, Korean Patent Publication No. 2003-93025 discloses a method of increasing the content of isoflavones by soaking after soaking in soybeans. However, in the soaking of seeds, the content of isoflavones is increased to prepare the cheonggukjang. There is a problem that is used in a limited way.

Therefore, the present inventors illuminate for a predetermined time and a period of time at the time when the seed becomes thick in the pod of one of the top 4 nodes engrafting the fully developing lobe which is the time when the pod of the bean seed is formed (time R5). The present invention was completed by developing a method for increasing the isoflavone content of seeds through treatment.

In particular, the present invention is a method of producing mature soybean seed with increased isoflavones by the light treatment during the growth of soybeans, the seed vitality is maintained, long-term storage is possible and can be easily used for the processing of most legume products.

Accordingly, the present invention is to provide a method for increasing the isoflavone content through the light treatment on the soybean growing.

Moreover, this invention provides the soybean seed obtained by the said method, and the soybean processed food using this seed.

The present invention is characterized by a method of increasing the isoflavone content through the light treatment on growing soybeans.

In the present invention, a method of increasing the isoflavone content of soybean seeds using a light treatment for the first time in soybean was developed. The method for increasing the isoflavone content, which is a representative functional ingredient of the soybeans provided in the present invention, can be very useful for producing soybean seeds with increased isoflavone content, thereby increasing the isoflavone content of soybean. Production of soybean seeds can facilitate processing into foods and foods fortified with isoflavone functional ingredients.

Hereinafter, the present invention will be described in more detail.

In the present invention, during the growing period of the beans in the pods of one of the top 4 nodes engrafting the complete development leaf at a time when the seeds are enlarged by treating the light for a certain time to keep the beans continuously exposed to light A method for increasing the isoflavone content.

Method for increasing the isoflavone content of soybean seed according to the present invention is as follows.

As a method for increasing the content of isoflavones according to the present invention, the soybean is exposed to light by light treatment at the time of soybean growth. The lighting is handled at the time of escalation (R5). More preferably, at 20 to 50 lux intensity, 1 to 4 hours before and after sunrise and 1 to 3 hours before and after sunset are treated for 10 to 30 days from the R5 time. In addition, lighting may be used to affect the flowering and narrowing of soybeans, such as fluorescent, incandescent, mercury, sodium, etc., but most preferably incandescent lamps that emit light of continuous wavelengths. If the lighting condition is out of the condition, the isoflavone content does not increase significantly, which is not preferable.

Soybean that can be applied to the present invention is soybean ( Glycine max L.) or a variant thereof, but is not limited thereto. Most preferably soybeans ( Glycine max L.).

The present invention can increase the isoflavone content of the soybeans by 12 to 40% compared to the normal grown soybeans through the additional light treatment in growing soybeans, so that the seed of the functional soybeans containing the isoflavones fixed to the seed at a high content It can be used to make isoflavone-containing soybean processed foods.

That is, the method of specifically increasing the isoflavone content by the light treatment provided in the present invention provides a method of fixedly increasing the isoflavone content of mature seeds rather than a temporary increase in the seed content of which the isoflavone content is increased. It provides a method that can be used to produce functional products using this seed.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited to the following examples.

Reference Example  1: Experimental Materials and Reagents

Soybeans used in the present invention used the Chongja No. 3, Daewon and Seonbean varieties, which were harvested from the cultivated packaging to maintain the unique characteristics of the varieties in the functional cropping department of the National Institute of Crop Science, Rural Development Administration. It can be used for soybean varieties with similar growth characteristics and ecotypes except for these varieties. Standard reagents for soy isoflavone analysis were purchased from Sigma (St. Luis, USA) with genistin, genistein, diidzein and glycidin, and daizin. ), Glycidin, acetyl daidzin, acetyl glycitin, acetyl genistin, malonyl daidzin, malonyl glycidine ( malonyl glycitin and malonyl genistin were purchased from LC Labs (Woburn, MA, USA). In addition, methanol, ethanol, water and acetonitrile for HPLC used in the analysis were J.T. Purchased from Baker (Phillipsburg, NJ, USA) and Sep-Pak cartridge was purchased from Water (Milford, MA, USA).

Reference Example  2: Sowing and Seed Growth

Bean varieties used for the light treatment provided by the present invention has a stage to be sown June 13 for normal germination and nutrient growth, seeds can be sown at the time soybean is sown in Korea. In order to cultivate soybeans, fertilization and management of soybeans were performed according to standard soybean growing methods.

In the stage of investigating seed germination, nutrient growth, flowering, narrowing, etc., the method of expressing the growth time of soybeans (Fehr & Carviness), which is a method of determining the growth time of certified soybeans for the unification of research and expression , 1977), soybean growth was investigated and used as a standard for light treatment.

[Table 1]

Expression of the development time, symbol and development state of the soybean used for the investigation of the growth time of soybean used in the present invention

Nutrition Reproductive Growth Development time Abbreviation Developmental status Development time Abbreviation Developmental status Germination VE When cotyledons appear on the ground bloom R1 When the first flowers bloomed on the main stem Cotyledon CV When the vegetation is developing Flowering R2 When we bloomed in one of the top two nodes which wore full development leaf Herb leaf V1 When the vegetation is fully developed At the time of narrowing R3 When you see a pod that reaches 5mm in one of the top 4 nodes of the fully developed lobe First biplane V2 When fully developed until the first biplane Narrowness R4 When you see a pod that reaches 2cm from one of the top 4 nodes of the fully developed leaf 2nd biplane V3 When fully developed until the second biplane Waiting for mouth R5 When the seed reaches 3mm in the pod of one of the top 4 nodes of the fully developed lobe .
.
.
.
.
.
. .
.
.
. Hypertrophy R6 When the beans in the pod of one of the top 4 nodes that developed the full development lobe were filled with green beans Maturity R7 When one of the normal pods that lived on the main stem showed aging (N-1) biplane Vn Fully developed to the (n-1) biplane Maturity R8 When 5% of pods are mature

Example  1: Bean varieties selection and light treatment

Soybean used in the present invention is a varieties corresponding to the middle-sized varieties and middle-sized varieties of varieties cultivated in Korea in the ecological classification of soybean (Lee Jae-eun, 2008) as shown in the following Table 3 Cheongja No. 3, Daewon soybeans, Seonyu beans, etc. This was used. Soybean cultivation was carried out based on the RDA standard soybean cultivation method after sowing on June 13, and fertilization was also applied according to the same criteria.

[Table 3]

Bean varieties used in the present invention as beans registered as varieties

Breed name Growth year Collection country (country of origin) Type Celadon No. 3 2004 Republic of Korea Glycine max L. Daewon Bean 1997 〃 〃 Soybean 2004 〃 〃

In consideration of the different growth times of the beans varieties, the experiment was performed by installing a light shielding facility and a light that is not irradiated to the adjacent beans during the light treatment of the beans varieties as shown in FIG.

The breeding time was examined for light treatment according to the growing season between varieties, and R5 was different for each breed so that the sun was different from 4 o'clock until 30 o'clock after sunrise (6 o'clock) in September and 18 o'clock before sunset (18:30). The illumination process was performed until 20:00. Lighting is typically used incandescent lamps for crops to respond to light, installed above 3 m above the ground, the distance between the light and the illumination is adjusted to 1.5 x 1.0 m and illuminance measured from 1 m above the ground 25 ~ 35 The lux light was treated to evenly transmit the beans.

[Table 2]

Light treatment start date (R5 time) and light treatment end date and time of the beans used in the present invention

Breed name Lighting start date Lighting end date Celadon No. 3 8/20 9/9 Daewon Bean 8/19 9/8 Soybean 8/11 8/31

Example  2: Extracting and Quantifying Isoflavones from Soybeans

HPLC instrument for soy isoflavones harvested in Example 1 is an Agilent 1100 series model equipped with a quaternary pump, an auto sampler, and a diode-array detector Was used.

For the quantitative analysis of isoflavone content in soybean, standard calibration curve and standard HPLC analysis conditions were established using standard materials, and isoflavones were extracted from bean seeds. Using the 12 kinds of isoflavones secured for qualitative and quantitative analysis of isoflavones, HPLC optimum separation conditions were first established, and various solvent combinations and solvent gradient conditions were established to establish optimum separation conditions. Is a solvent combination of solvent A (0.1% acetic acid H ₂ O) and solvent B (0.1% acetic acid acetonitrile) (0% B; 0min, 10% B: 20min, 20% B; 30min, 25% Gradient of solvent over time was set as B: 40min, 35% B; 50min, 40% B; 60min, 45% B; 70min, 50%). In addition, absorbance was measured at 260 nm with a diode array detector for isoflavone detection. The column for separation was a C-18 reversed phase column and the column temperature was measured at 30 ℃. And 12 isoflavones for quantitative analysis of soybean seed isoflavones (genistin, genistein, daidzein, glycidin, didzin, glycidine ( glycitin, acetyl daidzin, acetyl glycitin, acetyl genistin, malonyl daidzin, malonyl glycitin, malonyl HPLC standard calibration for malonyl genistin was performed by dissolving 1 to 2 mg of 12 isoflavones in methanol (200, 100, 80, 60, 40, 20, 10, 8, 6, 3, 1, 0.5, 0.25, 0.125, 0.06 μg / mL) was used to prepare a calibration curve by HPLC analysis.Isoflavone extraction of soybeans The optimum condition was the most isoflavone when extracted for 12 hours at 30 ° C. with 50% MeOH solvent. This was detected and established as the basic condition for material extraction.

As a result of measuring the isoflavone content after freeze-drying the soybeans by the growth period of Cheongja 3, the isoflavone content increased similarly to the R5-time light treatment and no treatment until the harvest time of the soybeans 4 as shown in FIG. Afterwards, the isoflavone content increased rapidly during the R5 season light treatment and mature seeds. In case of Daewon soybeans, isoflavones accumulate rapidly during the R5 light treatment after the harvest of foot beans 3. For soybeans, the isoflavone content of both treatments increased sharply after the harvesting of the soybeans 2, and the untreated treatment showed a modest increase after the harvesting of the soybeans 3. Showed.

In addition, the analysis results of the isoflavone content of each of the dried seeds after the completion of maturation and light treatment was 40 compared to the isoflavone content of 2,605 ug / g untreated by the light treatment as shown in Table 4 below The percentage increased to 3,647 ug / g of isoflavones, 12% for Daewon soybean, and 30% for isoflavones compared to non-treated soybeans.

[Table 4]

Isoflavone Contents of Mature Seeds Treated by Varieties

Division (ug / g) Celadon No. 3 Daewon Bean Soybean No treatment 2605.1 3337.3 2108.6 R5 3647.3 3749.9 2747.6

1 shows step by step from sowing of the beans to harvesting.

2 is a facility for soybean light treatment.

3 is an isoflavone chemical structure isolated from soybeans.

Figure 4 shows the variation of isoflavone content of the growth period and mature seed of Cheongja 3.

Figure 5 shows the variation of isoflavone content of the growing season and mature seeds of Daewon beans.

Figure 6 shows the variation of the isoflavone content of the growth period and mature seeds of the soybean.

Claims (8)

Illuminated during the growing season of the soybeans, so that the soybean plants are exposed to light, but sunrise is 10 to 30 days a day from the time when the seeds grow in the pods of one of the top 4 nodes of the full developing leaf (R5). Method for increasing the isoflavone content of soybean seed, characterized in that it is lighted 1 to 4 hours before and after and 1 to 3 hours before and after sunset. delete 2. A method according to claim 1, characterized by the treatment of illumination of 20 to 50 lux intensity. delete According to claim 1, wherein the soybean ( Glycine) max L.). Soybean seeds obtained by the method of any one of claims 1, 3 and 5. 7. Soybean seed according to claim 6, characterized in that the isoflavone content is increased by 12-40%. Soybean processed foods using the soybean seed of claim 6.

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