WO2013077529A1 - Methods for the processing of germinated soy germ with enhanced nutritive value and increased bioactive substance by using separated soy germ - Google Patents

Methods for the processing of germinated soy germ with enhanced nutritive value and increased bioactive substance by using separated soy germ Download PDF

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WO2013077529A1
WO2013077529A1 PCT/KR2012/005892 KR2012005892W WO2013077529A1 WO 2013077529 A1 WO2013077529 A1 WO 2013077529A1 KR 2012005892 W KR2012005892 W KR 2012005892W WO 2013077529 A1 WO2013077529 A1 WO 2013077529A1
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soy germ
germ
soy
germination
germinated
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PCT/KR2012/005892
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French (fr)
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Sun Lim Kim
Young Up KWON
Wook Han KIM
Choon Ki Lee
Jae Eun Lee
Yul Ho Kim
Mi Jung Kim
Yu Young Lee
Gun Ho JUNG
Dae Wook Kim
Jang Hoon Sung
Suk Chul HEO
Gun Yong Park
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Republic Of Korea (Management : Rural Development Administration)
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/70—Germinated pulse products, e.g. from soy bean sprouts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/01—Pulses or legumes in form of whole pieces or fragments thereof, without mashing or comminuting
    • A23L11/03—Soya beans, e.g. full-fat soya bean flakes or grits
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/40—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by drying or kilning; Subsequent reconstitution
    • A23L3/44—Freeze-drying
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/175—Amino acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/02—Acid
    • A23V2250/06—Amino acid
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A23V2250/2116—Flavonoids, isoflavones
    • A23V2250/21172—Soy Isoflavones, daidzein, genistein

Definitions

  • the present invention relates to a method for preparing germinated soy germ by germinating soy germ separated from soybeans.
  • the method of the present invention is characterized by comprising the steps of: drying soybean as a raw material in a hot-air dryer at 35°C for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ from soybeans; pulverizing the dried soybean seeds using a peeler; separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm; putting the separated soy germ into a mesh bag having a size of 40 ⁇ 50 cm and inducing the germination thereof using running water at about 20°C for 20 to 24 hours; harvesting the germinated soy germ with nutritional enhancement and cooking them with steam for 20 minutes; and freeze-drying or hot-air drying the steamed soy germ.
  • the germinated soy germ prepared according to the method of the present invention can be effectively used as a raw material for various foods, functional, industrial and medical materials. Further, the method of the present invention is effective to improve the usefulness of soybeans that are Korean traditional food materials and increase the applicability thereof as a new food material being helpful to the improvement in national public health.
  • Soybeans one of most favored Korean traditional foods, are rich in proteins and fats and contain various functional ingredients such as isoflavones, oligosaccharides and fibrins.
  • Soybean seeds are morphologically composed of a seed coat, a cotyledon and an embryo, wherein the cotyledon and soy germ are covered with the seed coat.
  • the embryo consists of epicotyl, hypocotyls and radical has a slightly embossed appearance.
  • the soybean seeds include 90 ⁇ 92% of the cotyledon, 6 ⁇ 8% of the seed coat, and 2% of the soy germ.
  • the soybean has been widely used as a processed food including tofu, soy milk, fermented soybeans, soy sauce, soybean paste, hot pepper paste, soybean sprouts and the like. Recently, as the study results for various bioactive substances included in the soybeans and processed soy foods have been reported, global market for processed soy foods has been remarkably growing.
  • isoflavones The representative example of bioactive substances contained in the soybeans is isoflavones. Due to the structurally similarity to estrogen, isoflavone is also called as phytoestrogen, and acts as an estrogen analogue in human body.
  • isoflavones show anti-cancer effects by inhibiting the growth of cancer cells such as prostate cancer, breast cancer, colon cancer, lung cancer and skin cancer, preventive effects of osteoporosis due to the lack of estrogen by increasing the absorption rate of potassium in the bone, bone density enhancing effects, anti-aging effects, anti-oxidant effects, lowering effects of the incidence rate of cardiovascular diseases or diabete mellitus.
  • estrogen was prescribed to about 15% of middle-aged women, but due to its side effects, soybeans as a natural food have been in the spotlight as a substitute.
  • Soybeans have high protein content of about 40%, and are thus called “beef being grown on a field”.
  • Food and Drug Administration made a decision based on the clinical study results being found till then, on that soy protein intake helps to prevent various cardiovascular diseases or alleviate their symptoms, and acknowledged the availability of soybeans and soy proteins.
  • the FDA now allows food manufacturers to make a health claim on a package if the product contains at least 6.25 g of soy protein per serving.
  • the health claim may say, “25 g of soy protein a day, as part of a diet low in saturated fat and cholesterol, may reduce the risk of heart diseases.”
  • soybeans have received attention as a source of vegetable proteins, and it has been found that various amino acids constituting the soy proteins exert a variety of bioregulatory function as well as nutritional function. It has been also reported that amino acids and peptides constituting the soy soybeans show various physiological activities such as in vivo anti-cancer and anti-tumor activities, hypotensive activity, serum cholesterol lowering activity, immunity improving activity and potassium absorption promoting activity.
  • GABA glutamic acid decarboxylase
  • GABA which is included in the brain of animals can induce the activation of metabolic function of brain cells, which results in showing improvement of scholastic skills and stress inhibition. Further, it has been reported that GABA is effective for the prevention or treatment of stroke, dementia and insomnia, attention improvement, and memory enhancement, and widely used in various hangover relieving beverages by activating liver function and stimulating alcohol metabolism.
  • oligosaccharides of the soybeans are low-calorie sweeteners and, in the large intestine, promote the growth of useful Enterobacteria including Lactobacillus bifidus .
  • Soy fibrins play an important role in normalization of bowel movement and are involved in the regulation of constipation and diarrhea, which results in reducing food passage time through the intestine.
  • dietary fibers and oligosaccharides included in the soybeans are effective to reduce the risk of developing colorectal cancer and other relating diseases, they are taking center stage by many researchers.
  • glucose, fructose, arabinose, pinitole and galactopinitole are present in a small amount, and the majorities are stachyose (1.4 ⁇ 4.1%) and raffinose (0.1 ⁇ 0.9%) called as oligosaccharides, that are normally provided in the form of a syrup.
  • soybeans In the soybeans, about 21 kinds of glycerolipids are present, and they serve as a framework for fatty acid synthesis. Soy lipids are composed of more than 5 different kinds of fatty acids that are commercially useful. Among them, the content and composition rate of palmitic acid (C16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and linolenic acid (18:3) influence on physical and chemical properties of fatty acids, and such fatty acids act as an crucial factor in determining the use of fatty acids. Especially, it has been found that unsaturated fatty acids of the soybeans have useful properties of preventing fats from excessively accumulating in the human body, removing cholesterol therefrom and making blood vessels relax.
  • the soybeans contain various bioactive substances as well as essential nutrients in a large quantity.
  • the bioactive substances and essential nutrients show variable change in their content depending on the region of soybean seeds.
  • the content of soy isoflavone is significantly higher in the soy germ than the seed coat or cotyledon.
  • Recent results of the USDA-ARS (Dr. Berhow) research have reported that the cotyledon contains 0.13 ⁇ 0.35% of isoflavone, and the soy germ contains 0.97 ⁇ 2.07% thereof.
  • the high contents of unsaturated fatty acid, isoflavone and soyasaponin in the soy germ are unfavorable to manufacture processed soy foods such as tofu and soy milk, and thus the manufacturer produce such processed soy foods with soybean ingredients after removing the seed coat and soy germ.
  • the manufacturer produce such processed soy foods with soybean ingredients after removing the seed coat and soy germ.
  • tofu and soy milk after removing the seed coat and soy germ from the soybeans there are several advantages of reducing soaking time, removing green or raw flavor, and improving chromacity (whiteness) of tofu (soy milk), leading to enhancement of commercial value and digestive power and diminishment of soybean-specific fishy smell and bitter taste.
  • the present inventors have endeavored to develop a new food and medical material which contains abundant nutrients and bioactive substances using soybean soy germ obtained as a by-product during the manufacture of processed soy products.
  • the present invention provides a method for preparing germinated soy germ, which includes the steps of:
  • soybeans as a raw material in a hot-air dryer at 35°C for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ therefrom;
  • freeze-drying or hot-air drying the steamed soy germ freeze-drying or hot-air drying the steamed soy germ.
  • the germinated soy germ according to the method of the present invention is obtained by germinating soy germ isolated from the soybeans for a shot time (20 ⁇ 24 hours), leading to enhancement of various nutritional functions.
  • the germinated soy germ are rich in bioactive substances such as free amino acids, gamma-aminobutyric acid (GABA), isoflavones and the like that have been regarded as a high functional ingredient in the academic world and the fields of food and medical industries, and thus can be effectively used as functional, industrial and medical materials as well as a raw material for various kinds of foods.
  • GABA gamma-aminobutyric acid
  • the germinated soy product that is prepared by separating soybean soy germ from soybean seeds, germinating the same to improve nutritional ingredients and bioactive substances, cooking the germinated soy germ with steam, and freeze-drying or hot-air drying the same, is an illustrative example for conversion of notion that has not been developed before now. Therefore, the germinated soy germ according to the present invention can be effectively used to improve the applicability of soybeans as a Korean traditional food material and be applied as a new food material helpful to enhance national health care.
  • Fig. 1 is a photograph showing the shape of soy germ isolated from soybean seeds.
  • Fig. 2 is a photograph showing the procedure of germinating the separated soy germ.
  • Fig. 3 is a photograph showing the change in shape of the separated soy germ at each time point of the germination.
  • Fig. 4 is a photograph showing the change in shape of the separated soy germ between before the germination and after the germination for 20 hours.
  • Fig. 5 is a graph showing the change in length, width and thickness of the separated soy germ at each time point of the germination.
  • Fig. 6 is graphs showing the change in real weight and dry weight of the separated soy germ at each time point of the germination.
  • Fig. 7 is a graph showing the change in contents of free amino acids and gamma-aminobutyric acid (GABA) in the separated soybean soy germ at each time point of the germination.
  • GABA gamma-aminobutyric acid
  • Fig. 8 is graphs showing the results of comparing the content of free amino acids of the separated soy germ between before the germination and after the germination for 20 hours.
  • Fig. 9 is a graph showing the change in contents of free sugar and oligosaccharide in the separated soy germ at each time point of the germination.
  • Fig. 10 is a graph showing the change in contents of fatty acid and unsaturated fatty acid in the separated soy germ at each time point of the germination.
  • Fig. 11 is a graph showing the change in contents of isoflavone in the separated soy germ at each time point of the germination.
  • Fig. 12 is HPLC chromatograms showing the results of comparing the content of isoflavones in the separated soy germ between before the germination and after the germination for 20 hours.
  • Fig. 13 is photographs showing the shape of soy germ that are germinated for 20 hours, cooked with steam and dried.
  • Fig. 14 is photographs showing the shape of soy germ that are germinated for 20 hours, cooked with steam and dried (free-drying and hot-air drying).
  • the present invention provides a method for preparing germinated soy germ by germinating soy germ separated from soybeans, which includes the steps of:
  • Soybeans used for the isolation of soy germ was “Daepungkong” (hereinafter, referred to as “soybean seeds”) which was fostered by the National Institute of Crop Science, Rural Development Administration on 2002 and cultivated thereby on 2010.
  • Fig. 1 is a photograph of soy germ isolated from the soybean seeds. Soybeans were dried in a hot-air dryer at 35°C for 72 hours and pulverized with a soybean peeler (Daeryuk Food Machine). At this time, the interval between blades of the soybean peeler was set to about 3 mm. As a result, the soybean seeds were isolated into a seed coat, 5 ⁇ 6 divided cotyledons and an soy germ. Here, the seed coat passed through the soybean peeler was removed by a ventilator installed within the peeler, simultaneously with harvesting the 5 ⁇ 6 divided cotyledons and soy germ. In order to isolate the soy germ from the mixture of cotyledons and soy germ, the mixture was sieved using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm.
  • Test Example 1 Seed water content depending on drying temperature, isolation level of seed coat and soy germ, and germination level of separated soy germ
  • Seed water content depending on drying temperature, isolation level of seed coat and soy germ, and germination state of separated soy germ of soybean seeds are shown in the following Table 1.
  • Seed water content (wt%) Isolation level of seed coat and soy germ Germination of soy germ Comparison 30°C 10.2 4 1 Drying for 72 hours at each temperature 35°C 9.5 2 1 40°C 8.7 2 3 45°C 7.9 2 4 50°C 7.4 1 5
  • Figs. 3, 4, 5 and 6 are photographs showing the germination level of soy germ and the change in weight thereof (real weight and dry weight).
  • the soy germ separated from soybeans showed the change in length, width and thickness as the germination was going on.
  • the length of soy germ increased from 4.27 mm before the germination to 7.78 mm after the germination for 24 hours, showing about 182% extension.
  • the width and thickness of soy germ were increased by about 119% and 159%, respectively, as compared to those before the germination.
  • Such phenomenon that the soy germ extended as the germination was going on suggest that although the soy germ are isolated from the soybean seeds, their germination can progress favorably.
  • the soy germ was physically cut during the isolation, their germination went successfully.
  • Fig. 5 shows the change in real weight (g) and dry weight (g) of the soy germ separated from soybeans during the time course of germination.
  • the weight of 100 soy germ was 638.0 mg, but their real weight rapidly increased as the germination was going on.
  • the real weight was 1357.9 mg and reached 1715.8 mg and 1912.1 mg, respectively after the germination for 20 and 24 hours.
  • the real weight increased by 269% and 300% as compared to that before the germination.
  • Such phenomenon is an important basis demonstrating that the separated soy germ are normally germinated. Further, considering the change in dry weight of the germinated soy germ, more positive results can be established.
  • the dry weight of 100 soy germ was decreased to 463.3 mg, which corresponds to 72.6% based on the dry weight of soy germ before the germination. From these results, it has been found that the soy germ show about 27.4% of decrease in dry weigh. The reason for the decrease in dry weight might be that as the separated soy germ is germinated, they rapidly consume various nutritional ingredients such as carbohydrates preserved therein.
  • Fig. 9 shows the results of analyzing nutritional ingredients of soy germ, that are obtained by pulverizing the separated soy germ with a pulverizer (Brabender, Germany) and measuring the composition of crude proteins, crude fats, crude fibers and fatty acids, content of free amino acids, carbon/nitrogen ratio (C/N ratio), content of isoflavones and the like.
  • the content of crude proteins was measured using an automatic analyzer (Kjeltec 2400 auto analyzer, Foss Tecator, Sweden) according to the Kjeldahl method, that of crude fats was measured using a crude fat automatic extractor (Soxtherm automatic system, Gerhardt, Germany), and that of crude fibers was measured using a crude fiber automatic analyzer (Fibertec 2010 auto analyzer, Foss Tecator, Huddinge, Sweden) according to the method as described by Van Soest et al . (1991).
  • Test Example 2 Change in C/N ration during the time course of germination
  • Table 2 shows the change in the contents of crude proteins, crude fats and crude fibers, and carbon/nitrogen ratio of the soy germ depending on the time course of germination.
  • Carbon/nitrogen ratio is a very important indicator to show the ratio between carbohydrates and nitrogen compounds.
  • the C/N ratio of the soy germ exhibited a tendency to decrease as the germination was going on. Such a phenomenon is because that carbohydrate substances preserved during the early stage of the germination are more rapidly consumed than nitrogen compounds.
  • Figs. 7 and 8 show the change in total amount of free amino acids and GABA ( ⁇ -amino butyric acid) depending on the time course of germination.
  • the total amount of free amino acids was 6079.6 mg/100 g and 6089.6 mg/100 g, respectively, which showed about 142% increase as compared to that before the germination.
  • the soy germ before the germination included 26.5 mg/100 g of GABA as a functional amino acid
  • the content of GABA was increased in the soy germ depending on the time course of germination.
  • the content of GABA was 703.5 mg/100 g and 718.0 mg/100 g, respectively, and showed about 27-fold higher as compared to that before the germination.
  • GABA is an amino acid which is ubiquitously present in plants, and during the seed germination, GABA and carbon dioxide are generated in large quantities by the action of a GAD enzyme to L-glutamic acid.
  • GABA which is present in the brain of animals can induce activation of metabolic function of brain cells, which results in showing improvement of scholastic skills and stress inhibition.
  • GABA is effective for the prevention or treatment of stroke, dementia and insomnia, attention improvement, and memory enhancement, and widely used in various hangover relieving beverages by activating liver function and stimulating alcohol metabolism. Therefore, the germinated soy germ can be effectively used as a material for functional food and medical industries.
  • Quantitative analysis of free sugar and oligosaccharides was carried out using a HPLC refractive index detector (Waters 410 RI Detector, USA). After 1 g of a sample was taken from freeze-dried powders of the soy germ, 10 mL of an 80% ethanol solution was added thereto, and the resulting mixture was subjected to ultrasonic extraction at 30 °C for about 60 minutes. In order to remove impurities, the resulting extract was centrifuged in a thermostat centrifuge at 4 °C, 15,000 rpm for 15 minutes to separate a supernatant.
  • the supernatant was filtered with a 0.45 ⁇ m membrane filter, passed through a Sep-Pak NH 2 to remove dyes and impurities, and then injected to a HPLC.
  • the column used in sugar analysis was Supelco LC-NH 2 (4mm ⁇ 25cm) column, and the mobile phase was a mixture of acetonitrile and ultrapure water (75:25 v/v) which was injected into at a flow rate of 1.5 mL/min.
  • Test Example 3 Change in contents of free sugar and oligosaccharides depending on the time course of germination
  • Table 3 shows the change in contents of free sugar and oligosaccharides depending on the time course of germination.
  • GC used in this analysis was an Agilent 6890 FID system (HP Co., Wilmington, DE, USA), and a HP-Innowax capillary 30 m ⁇ 0.25mm ⁇ 0.25 ⁇ m film (cross linked polyethylene glycol) was used as an analysis column.
  • An initial temperature of GC was set at 150°C, its final temperature was set at 280°C, and the temperature was increased at a rate of 4°C /min.
  • nitrogen (N 2 ) was injected at a rate of 10 mL/min, and during the analysis, the temperature of an inlet and a detector was maintained at 250°C and 300°C, respectively.
  • a FAME mix standard C14-C22 (Supelco, Bellefonte, PA, USA) was used.
  • Test Example 4 Change in contents of fatty acid and unsaturated fatty acid depending on the time course of germination
  • Table 4 shows the change in content of fatty acid depending on the time course of germination.
  • Germination period Fatty acid (wt%) Myristic acid (C14:0) Palmitic acid (C16:0) Stearic acid (C18:0) Oleic acid (C18:1) Linoleic acid (C18:2) Linolenic acid (C18:3) 0 - 17.14 3.90 4.30 53.59 21.06 5 - 17.49 2.55 2.04 55.64 22.28 10 0.19 16.68 3.36 4.79 53.84 21.14 15 0.22 17.77 2.47 1.98 55.48 22.09 20 0.31 17.23 3.41 5.04 53.24 20.77 24 - 17.30 3.72 5.27 52.67 21.03 Soybean seed (Daepungkong) 0.12 12.32 3.54 21.32 53.97 8.73
  • the composition ratio of linoleic acid (C18:2) was the highest by 53.59%, that of linolenic acid (C18:3) was 21.06%, that of palmitic acid (C16:0) was 17.14%, that of oleic acid (C18:1) was 4.30%, and that of stearic acid (C18:0) was the lowest.
  • the composition of fatty acids was in order of linolenic acid > oleic acid > palmitic acid > stearic acid > myristic acid. There was a significant difference in the fatty acid composition between the soybean seeds and the separated soy germ.
  • Figs. 11 and 12 show the change in content of isolflvones depending on the time course of germination.
  • the total isoflavone content of the soybean seeds (Daepungkong) used in the present invention was 1380.2 ⁇ g/g, and their isoflavone was composed of 599.6 ⁇ g/g of daidzein, 136.6 ⁇ g/g of glycitein and 644.0 ⁇ g/g of genistein, which showed the composition of isoflavone in order of genistein > daidzein > glycitein.
  • the total isoflavone content of the soy germ isolated from the soybean seeds was 10386.0 ⁇ g/g, and their isoflavone was composed of 4511.2.6 ⁇ g/g of daidzein, 4560.1 ⁇ g/g of glycitein and 1314.7 ⁇ g/g of genistein.
  • the composition of isoflavones is in order of glycitein > daidzein > genistein.
  • the total isoflavone content of the soy germ was about 7.5-fold higher than that of the soybean seeds, and in terms of the composition, daidzein was 7.5-fold higher, glycitein was 35.8-fold higher, and genistein was 1.3-fold higher in the soy germ as compared with the soybean seeds.
  • the total isoflavone content of the germinated soy germ showed about 115 ⁇ 128% increase after the germination for 20 ⁇ 24 hours as compared with the soy germ before the germination, and the content of genistein known as a functional ingredient was increased by about 123 ⁇ 139%.
  • the separated soy germ were germinated using running water for 20 ⁇ 24 hours, and the germinated soy germ were cooked with steam for 20 minutes.
  • the cooked soy germ were frozen in a -70°C ultra-low temperature refrigerator, followed by freeze-drying using a freeze-dryer or drying using a hot-air blower at 80°C (Figs. 13 and 14). If the thus dried product was powderized according to its intended purpose, it is possible to produce germinated soy germ products that are applicable as a food material.

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Abstract

The present invention relates to a method for preparing germinated soy germ by germinating soy germ separated from soybeans. In particular, the method of the present invention includes the steps of: drying soybeans as a raw material in a hot-air dryer at 35℃ for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ from the soybeans; pulverizing the dried soybean seeds using a peeler; separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm; putting the separated soy germ into a mesh bag having a size of 40 × 50 cm and inducing the germination thereof using running water at about 20℃ for 20 to 24 hours; harvesting the germinated soy germ with improvement in nutritional ingredients and cooking them with steam for 20 minutes; and freeze-drying or hot-air drying the steamed soy germ. The germinated soy germ prepared according to the method of the present invention can be effectively used as functional, industrial and medical materials as well as a raw material for various foods. Further, the method of the present invention is effective to improve the usefulness of soybeans that are Korean traditional food materials and increase the applicability thereof as a new food material being helpful to the improvement in national public health.

Description

METHODS FOR THE PROCESSING OF GERMINATED SOY GERM WITH ENHANCED NUTRITIVE VALUE AND INCREASED BIOACTIVE SUBSTANCE BY USING SEPARATED SOY GERM
The present invention relates to a method for preparing germinated soy germ by germinating soy germ separated from soybeans. In particular, the method of the present invention is characterized by comprising the steps of: drying soybean as a raw material in a hot-air dryer at 35℃ for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ from soybeans; pulverizing the dried soybean seeds using a peeler; separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm; putting the separated soy germ into a mesh bag having a size of 40 × 50 cm and inducing the germination thereof using running water at about 20℃ for 20 to 24 hours; harvesting the germinated soy germ with nutritional enhancement and cooking them with steam for 20 minutes; and freeze-drying or hot-air drying the steamed soy germ. The germinated soy germ prepared according to the method of the present invention can be effectively used as a raw material for various foods, functional, industrial and medical materials. Further, the method of the present invention is effective to improve the usefulness of soybeans that are Korean traditional food materials and increase the applicability thereof as a new food material being helpful to the improvement in national public health.

Soybeans, one of most favored Korean traditional foods, are rich in proteins and fats and contain various functional ingredients such as isoflavones, oligosaccharides and fibrins.
Soybean seeds are morphologically composed of a seed coat, a cotyledon and an embryo, wherein the cotyledon and soy germ are covered with the seed coat. The embryo (soy germ) consists of epicotyl, hypocotyls and radical has a slightly embossed appearance. The soybean seeds include 90~92% of the cotyledon, 6~8% of the seed coat, and 2% of the soy germ.
The soybean has been widely used as a processed food including tofu, soy milk, fermented soybeans, soy sauce, soybean paste, hot pepper paste, soybean sprouts and the like. Recently, as the study results for various bioactive substances included in the soybeans and processed soy foods have been reported, global market for processed soy foods has been remarkably growing.
The representative example of bioactive substances contained in the soybeans is isoflavones. Due to the structurally similarity to estrogen, isoflavone is also called as phytoestrogen, and acts as an estrogen analogue in human body.
Many researches have been focused on studying physiological activities and functionalities of isoflavones. It has been reported that isoflavones show anti-cancer effects by inhibiting the growth of cancer cells such as prostate cancer, breast cancer, colon cancer, lung cancer and skin cancer, preventive effects of osteoporosis due to the lack of estrogen by increasing the absorption rate of potassium in the bone, bone density enhancing effects, anti-aging effects, anti-oxidant effects, lowering effects of the incidence rate of cardiovascular diseases or diabete mellitus. In the U.S., estrogen was prescribed to about 15% of middle-aged women, but due to its side effects, soybeans as a natural food have been in the spotlight as a substitute.
Soybeans have high protein content of about 40%, and are thus called “beef being grown on a field”. On October 1999 in the U.S., the Food and Drug Administration (FDA) made a decision based on the clinical study results being found till then, on that soy protein intake helps to prevent various cardiovascular diseases or alleviate their symptoms, and acknowledged the availability of soybeans and soy proteins. The FDA now allows food manufacturers to make a health claim on a package if the product contains at least 6.25 g of soy protein per serving. The health claim may say, “25 g of soy protein a day, as part of a diet low in saturated fat and cholesterol, may reduce the risk of heart diseases.”
The soybeans have received attention as a source of vegetable proteins, and it has been found that various amino acids constituting the soy proteins exert a variety of bioregulatory function as well as nutritional function. It has been also reported that amino acids and peptides constituting the soy soybeans show various physiological activities such as in vivo anti-cancer and anti-tumor activities, hypotensive activity, serum cholesterol lowering activity, immunity improving activity and potassium absorption promoting activity.
In particular, during the germination of seeds, glutamic acid decarboxylase (GAD) catalyzes the decarboxylation of L-glutamic acid to form a large quantity of GABA as a functional amino acid and carbon dioxide (CO2).
GABA which is included in the brain of animals can induce the activation of metabolic function of brain cells, which results in showing improvement of scholastic skills and stress inhibition. Further, it has been reported that GABA is effective for the prevention or treatment of stroke, dementia and insomnia, attention improvement, and memory enhancement, and widely used in various hangover relieving beverages by activating liver function and stimulating alcohol metabolism.
It has been reported that oligosaccharides of the soybeans are low-calorie sweeteners and, in the large intestine, promote the growth of useful Enterobacteria including Lactobacillus bifidus. Soy fibrins play an important role in normalization of bowel movement and are involved in the regulation of constipation and diarrhea, which results in reducing food passage time through the intestine. Recently, as several research results have been reported that dietary fibers and oligosaccharides included in the soybeans are effective to reduce the risk of developing colorectal cancer and other relating diseases, they are taking center stage by many researchers. Generally, among soluble carbohydrates included in the soybean seeds, glucose, fructose, arabinose, pinitole and galactopinitole are present in a small amount, and the majorities are stachyose (1.4~4.1%) and raffinose (0.1~0.9%) called as oligosaccharides, that are normally provided in the form of a syrup.
In the soybeans, about 21 kinds of glycerolipids are present, and they serve as a framework for fatty acid synthesis. Soy lipids are composed of more than 5 different kinds of fatty acids that are commercially useful. Among them, the content and composition rate of palmitic acid (C16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and linolenic acid (18:3) influence on physical and chemical properties of fatty acids, and such fatty acids act as an crucial factor in determining the use of fatty acids. Especially, it has been found that unsaturated fatty acids of the soybeans have useful properties of preventing fats from excessively accumulating in the human body, removing cholesterol therefrom and making blood vessels relax.
As described above, the soybeans contain various bioactive substances as well as essential nutrients in a large quantity. According to the results reported until now, the bioactive substances and essential nutrients show variable change in their content depending on the region of soybean seeds. For example, the content of soy isoflavone is significantly higher in the soy germ than the seed coat or cotyledon. Recent results of the USDA-ARS (Dr. Berhow) research have reported that the cotyledon contains 0.13~0.35% of isoflavone, and the soy germ contains 0.97~2.07% thereof.
However, the high contents of unsaturated fatty acid, isoflavone and soyasaponin in the soy germ are unfavorable to manufacture processed soy foods such as tofu and soy milk, and thus the manufacturer produce such processed soy foods with soybean ingredients after removing the seed coat and soy germ. For example, in case of manufacturing tofu and soy milk after removing the seed coat and soy germ from the soybeans, there are several advantages of reducing soaking time, removing green or raw flavor, and improving chromacity (whiteness) of tofu (soy milk), leading to enhancement of commercial value and digestive power and diminishment of soybean-specific fishy smell and bitter taste. Further, by-products generated during the manufacture of agricultural products including the soybeans cause serious social problems in that the cost of discarding their waste material is very high, and such waste material can cause various environmental pollutions. Thus, developing a new material using such by-products can be effective to reduce social cost for waste material disposal and minimize environmental pollutions.
Therefore, the present inventors have endeavored to develop a new food and medical material which contains abundant nutrients and bioactive substances using soybean soy germ obtained as a by-product during the manufacture of processed soy products.
It is an object of the present invention to provide a method for preparation of germinated soy germ with improvement in various nutritional ingredients, free amino acids, GABA, isoflavones and the like, which includes the steps of separating soybean soy germ from soybean seeds, and germinating the separated soy germ.

In accordance with the first aspect thereof, the present invention provides a method for preparing germinated soy germ, which includes the steps of:
drying soybeans as a raw material in a hot-air dryer at 35℃ for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ therefrom;
pulverizing the dried soybean seeds using a peeler;
separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm;
putting the separated soy germ into a mesh bag having a size of 40 × 50 cm and inducing the germination thereof using running water at about 20℃ for 20 to 24 hours;
harvesting the germinated soy germ with improvement in nutritional ingredients and cooking them with steam for 20 minutes; and
freeze-drying or hot-air drying the steamed soy germ.
The germinated soy germ according to the method of the present invention is obtained by germinating soy germ isolated from the soybeans for a shot time (20~24 hours), leading to enhancement of various nutritional functions. Especially, the germinated soy germ are rich in bioactive substances such as free amino acids, gamma-aminobutyric acid (GABA), isoflavones and the like that have been regarded as a high functional ingredient in the academic world and the fields of food and medical industries, and thus can be effectively used as functional, industrial and medical materials as well as a raw material for various kinds of foods.
In addition, according to the present invention, the germinated soy product that is prepared by separating soybean soy germ from soybean seeds, germinating the same to improve nutritional ingredients and bioactive substances, cooking the germinated soy germ with steam, and freeze-drying or hot-air drying the same, is an illustrative example for conversion of notion that has not been developed before now. Therefore, the germinated soy germ according to the present invention can be effectively used to improve the applicability of soybeans as a Korean traditional food material and be applied as a new food material helpful to enhance national health care.
Fig. 1 is a photograph showing the shape of soy germ isolated from soybean seeds.
Fig. 2 is a photograph showing the procedure of germinating the separated soy germ.
Fig. 3 is a photograph showing the change in shape of the separated soy germ at each time point of the germination.
Fig. 4 is a photograph showing the change in shape of the separated soy germ between before the germination and after the germination for 20 hours.
Fig. 5 is a graph showing the change in length, width and thickness of the separated soy germ at each time point of the germination.
Fig. 6 is graphs showing the change in real weight and dry weight of the separated soy germ at each time point of the germination.
Fig. 7 is a graph showing the change in contents of free amino acids and gamma-aminobutyric acid (GABA) in the separated soybean soy germ at each time point of the germination.
Fig. 8 is graphs showing the results of comparing the content of free amino acids of the separated soy germ between before the germination and after the germination for 20 hours.
Fig. 9 is a graph showing the change in contents of free sugar and oligosaccharide in the separated soy germ at each time point of the germination.
Fig. 10 is a graph showing the change in contents of fatty acid and unsaturated fatty acid in the separated soy germ at each time point of the germination.
Fig. 11 is a graph showing the change in contents of isoflavone in the separated soy germ at each time point of the germination.
Fig. 12 is HPLC chromatograms showing the results of comparing the content of isoflavones in the separated soy germ between before the germination and after the germination for 20 hours.
Fig. 13 is photographs showing the shape of soy germ that are germinated for 20 hours, cooked with steam and dried.
Fig. 14 is photographs showing the shape of soy germ that are germinated for 20 hours, cooked with steam and dried (free-drying and hot-air drying).
Hereinafter, the present invention will be described in more detail.
In accordance with an aspect, the present invention provides a method for preparing germinated soy germ by germinating soy germ separated from soybeans, which includes the steps of:
(1) employing soybeans that are cultivated in South Korea on 2010 (Daepungkong);
(2) drying the soybeans as a raw material in a hot-air dryer at 35℃ for 72 hours so as to efficiently isolate seed coats, cotyledons and soy germ from the soybeans and maintain optimal germination capacity thereof;
(3) pulverizing the dried soybean seeds using a peeler;
(4) separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm;
(5) putting the separated soy germ into a mesh bag having a size of 40 × 50 cm and inducing the germination thereof using running water at about 20℃ for 20 to 24 hours;
(6) analyzing nutritional ingredients of the soy germ during the germination depending on the time course (before the germination, 5, 10, 15, 20 and 24 hours after the germination);
(7) collecting the germinated soy germ with improvement in nutritional ingredients after the germination for 20~24 hours;
(8) cooking the collected soy germ with steam for 20 minutes; and
(9) freeze-drying or hot-air drying at 80℃ the steamed soy germ.

The present invention is further illustrated by the following examples. However, it shall be understood that these examples are only used to specifically set forth the present invention, rather than being understood that they are used to limit the present invention in any form.

[Example 1]
Soybeans used for the isolation of soy germ was “Daepungkong” (hereinafter, referred to as “soybean seeds”) which was fostered by the National Institute of Crop Science, Rural Development Administration on 2002 and cultivated thereby on 2010.
Fig. 1 is a photograph of soy germ isolated from the soybean seeds. Soybeans were dried in a hot-air dryer at 35℃ for 72 hours and pulverized with a soybean peeler (Daeryuk Food Machine). At this time, the interval between blades of the soybean peeler was set to about 3 mm. As a result, the soybean seeds were isolated into a seed coat, 5~6 divided cotyledons and an soy germ. Here, the seed coat passed through the soybean peeler was removed by a ventilator installed within the peeler, simultaneously with harvesting the 5~6 divided cotyledons and soy germ. In order to isolate the soy germ from the mixture of cotyledons and soy germ, the mixture was sieved using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm.

Test Example 1: Seed water content depending on drying temperature, isolation level of seed coat and soy germ, and germination level of separated soy germ
Seed water content depending on drying temperature, isolation level of seed coat and soy germ, and germination state of separated soy germ of soybean seeds are shown in the following Table 1.
Drying temperature Seed water content (wt%) Isolation level of seed coat and soy germ Germination of soy germ Comparison
30℃ 10.2 4 1 Drying for 72 hours at each temperature
35℃ 9.5 2 1
40℃ 8.7 2 3
45℃ 7.9 2 4
50℃ 7.4 1 5
Note) 1: very good, 2: good, 3: normal, 4: bad, 5: very bad

Considering the results obtained at each drying temperature, the higher the drying temperature is, the more the isolation of seed coats and soy germ goes on smoothly. However, because the isolated soy germ was poorly germinated at higher drying temperature, it is preferable to dry soybean seeds at 35 ℃ for 72 hours for the isolation of seed coat (hull), cotyledon and soy germ and the germination of separated soy germ.

[Example 2]
As shown in Fig. 2, 6 kg of the soy germ isolated from soybean seeds were put into a mesh bag having a size of 40 × 50 cm and subjected to germination using running water. At this time, in order to monitor the change in nutritional ingredients of the germinated soy germ, samples were collected at each time point (before the germination, 5, 10, 15, 20 and 24 hours after the germination). The reason that running water is used to induce the germination of soy germ is to efficiently remove by-products eluted from the soy germ during the germination, for example, gas being generated by vigorous respiration of the germinated soy germ.

[Example 3]
Figs. 3, 4, 5 and 6 are photographs showing the germination level of soy germ and the change in weight thereof (real weight and dry weight).
As shown in Figs. 3, 4 and 5, the soy germ separated from soybeans showed the change in length, width and thickness as the germination was going on. Especially, the length of soy germ increased from 4.27 mm before the germination to 7.78 mm after the germination for 24 hours, showing about 182% extension. The width and thickness of soy germ were increased by about 119% and 159%, respectively, as compared to those before the germination. Such phenomenon that the soy germ extended as the germination was going on suggest that although the soy germ are isolated from the soybean seeds, their germination can progress favorably. In particular, as shown in Fig. 3, although the soy germ was physically cut during the isolation, their germination went successfully.
Fig. 5 shows the change in real weight (g) and dry weight (g) of the soy germ separated from soybeans during the time course of germination. Before the germination, the weight of 100 soy germ was 638.0 mg, but their real weight rapidly increased as the germination was going on. After the germination for 5 hours, the real weight was 1357.9 mg and reached 1715.8 mg and 1912.1 mg, respectively after the germination for 20 and 24 hours. The real weight increased by 269% and 300% as compared to that before the germination. Such phenomenon is an important basis demonstrating that the separated soy germ are normally germinated. Further, considering the change in dry weight of the germinated soy germ, more positive results can be established. After the germination for 24 hours, the dry weight of 100 soy germ was decreased to 463.3 mg, which corresponds to 72.6% based on the dry weight of soy germ before the germination. From these results, it has been found that the soy germ show about 27.4% of decrease in dry weigh. The reason for the decrease in dry weight might be that as the separated soy germ is germinated, they rapidly consume various nutritional ingredients such as carbohydrates preserved therein.

[Example 3]
1) Change in contents of crude proteins, crude fats and crude fibers, and carbon/nitrogen (C/N) ratio
Fig. 9 shows the results of analyzing nutritional ingredients of soy germ, that are obtained by pulverizing the separated soy germ with a pulverizer (Brabender, Germany) and measuring the composition of crude proteins, crude fats, crude fibers and fatty acids, content of free amino acids, carbon/nitrogen ratio (C/N ratio), content of isoflavones and the like.
The content of crude proteins was measured using an automatic analyzer (Kjeltec 2400 auto analyzer, Foss Tecator, Sweden) according to the Kjeldahl method, that of crude fats was measured using a crude fat automatic extractor (Soxtherm automatic system, Gerhardt, Germany), and that of crude fibers was measured using a crude fiber automatic analyzer (Fibertec 2010 auto analyzer, Foss Tecator, Huddinge, Sweden) according to the method as described by Van Soest et al. (1991).

Test Example 2: Change in C/N ration during the time course of germination
Table 2 shows the change in the contents of crude proteins, crude fats and crude fibers, and carbon/nitrogen ratio of the soy germ depending on the time course of germination.

Germination period Crude protein (wt%) Crude fat (wt%) Crude fiber (wt%) Carbon/nitrogen ratio
(C/N ratio)
0 35.1 10.1 3.08 8.2
5 36.7 9.9 3.08 8.1
10 37.0 9.8 3.09 8.1
15 37.7 9.7 3.17 8.1
20 38.2 9.7 3.21 8.0
24 38.7 9.7 3.20 7.8
Soybean seed
(Daepungkong)		 39.2 18.9 4.34 7.8

As shown in Fig. 9 and Table 2, the contents of crude proteins and crude fibers of the separated soy germ were slightly lower than those of soybean seeds (Daepungkong), but they were increased as the germination was going on. While the content of crude fats in the soybean seeds were 18.9%, that of crude fats in the soy germ was 10.1%, which tends downward.
Carbon/nitrogen ratio (C/N ratio) is a very important indicator to show the ratio between carbohydrates and nitrogen compounds. The C/N ratio of the soy germ exhibited a tendency to decrease as the germination was going on. Such a phenomenon is because that carbohydrate substances preserved during the early stage of the germination are more rapidly consumed than nitrogen compounds.

[Example 4]
2) Change in content of free amino acids
In order to analyze free amino acids, 10 mL of a 3% TCA (trichloroacetic acid) solution was added to 0.3 g of a sample, and the mixture was subjected to extraction for 1 hour with stirring. The resulting mixture was centrifuged at 15,000 rpm for 15 minutes, filtered with a Millipore 0.45 mL syringe filter (Milford, USA), and then used in a free amino acid analysis. The quantitative analysis of free amino acids was carried out using an Amino acid Auto-analyzer (Hitachi L-8800, Japan) which employed an ion exchange #2622SC-PF column. As a free amino acid standard, Type AN II and Type B (Wako, Wako-shi, Japan) were used.
Figs. 7 and 8 show the change in total amount of free amino acids and GABA (γ-amino butyric acid) depending on the time course of germination.
The total amounts of free amino acids and GABA in the soybean seeds were 6544.3 mg/100 g and 57.9 mg/100 g, respectively, but those of free amino acids and GABA in the separated soy germ were 4290.1 mg/100 g and 26.5 mg/100 g, respectively, these results suggest that the contents of free amino acids and GABA be higher in the soybean seeds than in the separated soy germ. However, in case of inducing the germination of separated soy germ, total amounts of free amino acids and GABA were increased as the germination was going on. After the germination for 20 and 24 hours, the total amount of free amino acids was 6079.6 mg/100 g and 6089.6 mg/100 g, respectively, which showed about 142% increase as compared to that before the germination. In particular, while the soy germ before the germination included 26.5 mg/100 g of GABA as a functional amino acid, the content of GABA was increased in the soy germ depending on the time course of germination. After the germination for 20 and 24 hours, the content of GABA was 703.5 mg/100 g and 718.0 mg/100 g, respectively, and showed about 27-fold higher as compared to that before the germination. It has been known in the art that GABA is an amino acid which is ubiquitously present in plants, and during the seed germination, GABA and carbon dioxide are generated in large quantities by the action of a GAD enzyme to L-glutamic acid. GABA which is present in the brain of animals can induce activation of metabolic function of brain cells, which results in showing improvement of scholastic skills and stress inhibition. Further, it has been known that GABA is effective for the prevention or treatment of stroke, dementia and insomnia, attention improvement, and memory enhancement, and widely used in various hangover relieving beverages by activating liver function and stimulating alcohol metabolism. Therefore, the germinated soy germ can be effectively used as a material for functional food and medical industries.

[Example 5]
Change in contents of free sugar and oligosaccharides
Quantitative analysis of free sugar and oligosaccharides was carried out using a HPLC refractive index detector (Waters 410 RI Detector, USA). After 1 g of a sample was taken from freeze-dried powders of the soy germ, 10 mL of an 80% ethanol solution was added thereto, and the resulting mixture was subjected to ultrasonic extraction at 30 ℃ for about 60 minutes. In order to remove impurities, the resulting extract was centrifuged in a thermostat centrifuge at 4 ℃, 15,000 rpm for 15 minutes to separate a supernatant. The supernatant was filtered with a 0.45㎛ membrane filter, passed through a Sep-Pak NH2 to remove dyes and impurities, and then injected to a HPLC. The column used in sugar analysis was Supelco LC-NH2 (4mm×25㎝) column, and the mobile phase was a mixture of acetonitrile and ultrapure water (75:25 v/v) which was injected into at a flow rate of 1.5 mL/min.

Test Example 3: Change in contents of free sugar and oligosaccharides depending on the time course of germination
Table 3 shows the change in contents of free sugar and oligosaccharides depending on the time course of germination.
Germination period Content of sugar (wt%)
Fructose Glucose Sucrose Galactose Raffinose Stachyose Total sugars
0 0.32 0.13 7.11 0.36 1.43 19.21 28.57
5 0.20 0.12 5.46 0.36 1.04 17.17 24.35
10 0.19 0.10 4.47 0.38 1.00 16.56 22.70
15 0.18 0.09 3.65 0.32 1.02 16.38 21.65
20 0.16 0.08 2.96 0.32 1.01 16.02 20.56
24 0.15 0.07 0.88 0.30 0.95 15.71 18.07
Soybean seed
(Daepungkong)		 0.18 0.11 4.21 0.09 0.93 3.91 9.43

Considering the content of sugar as shown in Table 3 and Fig. 9, while the total content of sugar before the soy germ germination was 28.57%, after the germination for 20~24 hours, it was decreased up to 20.56~18.07%. Sucrose whose content before the germination was 7.11% was decreased up to 2.96~0.88% after the germination for 20~24 hours, but the change in contents of fructose, glucose and galactose as a monosaccharide and raffinose and stachyose as an oligosaccharide was relatively minor.

[Example 6]
4) Change in fatty acid composition
For the analysis of fatty acids, a mixed solution of methanol : heptane : benzene : 2,2-dimethoxypropane : H2SO4 (37:36:20:5:2, v/v) was added to 0.5 g of a powder sample, and the resulting mixture was heated to 80℃ such that lipid degradation and transmethylation occurred simultaneously. After heating for about 1 hour, the reaction solution was cooled down, and a supernatant containing fatty acid methyl ester (FAME) was separated therefrom, followed by capillary GC (capillary gas chromatography). At this time, GC used in this analysis was an Agilent 6890 FID system (HP Co., Wilmington, DE, USA), and a HP-Innowax capillary 30 m × 0.25mm × 0.25㎛ film (cross linked polyethylene glycol) was used as an analysis column. An initial temperature of GC was set at 150℃, its final temperature was set at 280℃, and the temperature was increased at a rate of 4℃ /min. As a carrier gas, nitrogen (N2) was injected at a rate of 10 mL/min, and during the analysis, the temperature of an inlet and a detector was maintained at 250℃ and 300℃, respectively. A FAME mix standard (C14-C22) (Supelco, Bellefonte, PA, USA) was used.

Test Example 4: Change in contents of fatty acid and unsaturated fatty acid depending on the time course of germination
Table 4 shows the change in content of fatty acid depending on the time course of germination.
Germination period Fatty acid (wt%)
Myristic acid
(C14:0)		 Palmitic acid (C16:0) Stearic acid (C18:0) Oleic acid (C18:1) Linoleic acid
(C18:2)		 Linolenic acid
(C18:3)
0 - 17.14 3.90 4.30 53.59 21.06
5 - 17.49 2.55 2.04 55.64 22.28
10 0.19 16.68 3.36 4.79 53.84 21.14
15 0.22 17.77 2.47 1.98 55.48 22.09
20 0.31 17.23 3.41 5.04 53.24 20.77
24 - 17.30 3.72 5.27 52.67 21.03
Soybean seed
(Daepungkong)		 0.12 12.32 3.54 21.32 53.97 8.73
As shown in Table 4 and Fig. 10, in the separated soy germ, the composition ratio of linoleic acid (C18:2) was the highest by 53.59%, that of linolenic acid (C18:3) was 21.06%, that of palmitic acid (C16:0) was 17.14%, that of oleic acid (C18:1) was 4.30%, and that of stearic acid (C18:0) was the lowest. However, in the soybean seeds (Daepungkong), the composition of fatty acids was in order of linolenic acid > oleic acid > palmitic acid > stearic acid > myristic acid. There was a significant difference in the fatty acid composition between the soybean seeds and the separated soy germ. As shown in Figs. 10 and 11, considering the change in fatty acid composition of the separated soy germ depending on the time course of germination, there was no clear change in fatty acid composition. Further, the composition ratio of unsaturated fatty acids was slightly increased at the early stage of germination, but it was decreased depending on the time course of germination and reached 79.0% which was the same as that before the soy germ germination.

[Example 7]
5) Change in content of isoflavones
In order to analyze the content of isoflavones in the soybeans and soy germ thereif, 10 mL of a 1 N HCl solution was added to each of 1.0 g of pulverized soybean and soy germ samples, and the resulting mixture was subjected to hydrolysis in a 100℃ heating block (Thermolyne, USA) for 1 hour so as to induce the transform into aglycone. After adjusting its final volume to 50 mL with methanol, the resulting mixture was kept at room temperature for 24 hours. Some portion of the mixture was taken out, passed through a 0.45 μm PTFE syringe filter (Waters, Milford, MA, USA), and injected into a Waters e2695 HPLC equipped with a Waters 2996 PAD (Waters, Milford, MA, USA). The mobile phase was a mixture of acetonitrile and ultrapure water (35:65 v/v), which was injected into the HPLC at a flow rate of 20 μL/min. As a column, a YMC-Pack ODS-AM303 (4.6 × 250 mm) column (YMC Inc, Wilmington, NC) was employed, and a UV wavelength of a detector was 254 nm.
Figs. 11 and 12 show the change in content of isolflvones depending on the time course of germination.
The total isoflavone content of the soybean seeds (Daepungkong) used in the present invention was 1380.2 μg/g, and their isoflavone was composed of 599.6 μg/g of daidzein, 136.6 μg/g of glycitein and 644.0 μg/g of genistein, which showed the composition of isoflavone in order of genistein > daidzein > glycitein. However, the total isoflavone content of the soy germ isolated from the soybean seeds was 10386.0 μg/g, and their isoflavone was composed of 4511.2.6 μg/g of daidzein, 4560.1 μg/g of glycitein and 1314.7 μg/g of genistein. In contrast to the soybean seeds, the composition of isoflavones is in order of glycitein > daidzein > genistein. These results showed that there was significant difference in isofalvone content and composition between the soybean seeds and the soy germ. Especially, the total isoflavone content of the soy germ was about 7.5-fold higher than that of the soybean seeds, and in terms of the composition, daidzein was 7.5-fold higher, glycitein was 35.8-fold higher, and genistein was 1.3-fold higher in the soy germ as compared with the soybean seeds. These results suggest that the soy germ were rich in isoflavones.
Depending on the time course of germination, the total isoflavone content of the germinated soy germ showed about 115~128% increase after the germination for 20~24 hours as compared with the soy germ before the germination, and the content of genistein known as a functional ingredient was increased by about 123~139%.

[Example 8]
In order to improve the contents and function of various nutritional ingredients and bioactive substances and enlarge its applicability to various food materials, the separated soy germ were germinated using running water for 20~24 hours, and the germinated soy germ were cooked with steam for 20 minutes. The cooked soy germ were frozen in a -70℃ ultra-low temperature refrigerator, followed by freeze-drying using a freeze-dryer or drying using a hot-air blower at 80℃ (Figs. 13 and 14). If the thus dried product was powderized according to its intended purpose, it is possible to produce germinated soy germ products that are applicable as a food material. However, in case of cooking the germinated soy germ, there is a problem in that processing properties of the germinated soy germ can be deteriorated due to the degradation of proteins. Thus, according to the intended purpose, it is possible to freeze-dry or hot-air dry the germinated soy germ, followed by powderizing without cooking with steam.

Claims (7)

  1. A method for preparing germinated soy germ with improvement in nutritional function and increase in bioactive substances, comprising:
    isolating soy germ from soybeans; and
    inducing the germination of the soy germ separated above using running water.
  2. The method according to Claim 1, wherein the bioactive substance is free amino acids or gamma-aminobutyric acid (GABA).
  3. The method according to Claim 1, wherein the physiologically substance is isoflavone, genistein, glycitein, or daidzein.

  4. A method for preparing germinated soy germ with improvement in nutritional function and increase in bioactive substances using soybean soy germ, comprising:
    drying soybean seeds as a raw material;
    pulverizing the dried soybean seeds using a peeler;
    separating soy germ from the pulverized soybean seeds;
    inducing the germination of separated soy germ using running water;
    harvesting the germinated soy germ and cooking them with steam; and
    freeze-drying or hot-air drying the steamed soy germ.

  5. The method according to Claim 4, which comprises
    drying soybeans in a hot-air dryer at 35℃ for 72 hours;
    pulverizing the dried soybean seeds using a peeler;
    separating soy germ from the pulverized soybean seeds using a series of sieves having a sieve opening of 2.36 mm, 2.0 mm and 1.18 mm;
    inducing the germination of separated soy germ using running water at about 20℃ for 20 to 24 hours;
    harvesting the germinated soy germ and cooking them with steam for 20 minutes; and
    freeze-drying or hot-air drying the steamed soy germ.

  6. The method according to Claim 4, which is characterized by cooking the germinated soy germ with steam for 20 minutes and freeze-drying the steamed soy germ at -70℃.
  7. The method according to Claim 4, which is characterized by cooking the germinated soy germ with steam for 20 minutes and hot-air drying the steamed soy germ at 80℃.

PCT/KR2012/005892 2011-11-23 2012-07-24 Methods for the processing of germinated soy germ with enhanced nutritive value and increased bioactive substance by using separated soy germ WO2013077529A1 (en)

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KR1020110122965A KR101376229B1 (en) 2011-11-23 2011-11-23 Nutritional enhancement of the functionality of gemmule separated from bean and a fabricateion process of bioactive substance increased germinated gemmule
KR10-2011-0122965 2011-11-23

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KR20170054115A (en) * 2015-11-09 2017-05-17 대한민국(농촌진흥청장) A pharmaceutical composition comprising extract from germinated gemmule of bean for preventing or treating osteoporosis
KR101898261B1 (en) * 2018-07-10 2018-09-14 대한민국 A pharmaceutical composition comprising extract from germinated gemmule of bean for preventing or treating osteoporosis

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JP3144630B2 (en) * 1997-07-31 2001-03-12 株式会社サラダコスモ Processed soy food and method for producing the same
JP2002112723A (en) * 2000-10-06 2002-04-16 Ajinomoto Co Inc Method for producing soybean material with high embryo content

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KR100436024B1 (en) * 2002-08-22 2004-06-12 유치호 The ice cream containing isoflavone and the method thereof

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JP2002112723A (en) * 2000-10-06 2002-04-16 Ajinomoto Co Inc Method for producing soybean material with high embryo content

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106333339A (en) * 2016-09-06 2017-01-18 华南理工大学 Preparation method of crude soy sauce

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