JP2005095156A - Soybean extract, method for producing the same and use of the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a soybean extract having more protein (≥40 % in a solid) than regular broth, and to provide a method for producing the same. <P>SOLUTION: The soybean extract is obtained by hot water extraction of soybeans which is gradually subjected to hot water treatment and single-cell forming treatment. The soybeans to be used are dehulled soybeans. The hot water treatment comprises steaming the soybeans in hot water at 100-150°C for 5-60 min. The single-cell forming treatment comprises substantially forming the cells of the soybeans into single cells by physical treatment. The method for producing the soybean extract comprises subjecting the soybeans to hot water treatment followed by physical treatment to form into single cells, and subjecting the soybeans to hot water extraction in order to obtain the extract having more protein (≥40 % in a solid) than regular broth. An inhibitor of angiotensin converting enzyme comprising either one of the soybean extracts is also provided. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a soybean extract that can be effectively used as a food material, a production method thereof, and a use thereof.

  Boiled soybean soup is a by-product generated during the production of miso and natto, and has conventionally been a source of pollution as waste, but there are examples where it is used as a raw material for fermentation of lactic acid bacteria (Patent Documents 1 to 3). Soy boiled soy is effective as a fermentation material, but it is not widely used due to its unique soy odor.

Further, although a method for stepwise extraction of soybean is shown in Patent Document 4, it has not yet reached an efficient method for producing an intercellular binding component and its use.
Japanese Patent Publication No. 62-27777 JP-A-63-317055 JP-A-8-56651 JP 2002-256281 A

  An object of the present invention is to collect and effectively use components contained in soybean, particularly components containing intercellular binding substances. An object of this invention is to provide the component (40% or more in solid substance) with more protein than a normal boiled juice, and its manufacturing method.

The gist of the present invention is the soybean extract (1) to (4) below.
(1) A soybean extract obtained by subjecting soybeans to hydrothermal treatment and unicellular treatment in stages.
(2) The soybean extract as described in (1) above, using a molted soybean.
(3) The soybean extract as described in (1) or (2) above, wherein the hydrothermal treatment is to cook soybeans with hot water at 100 to 150 ° C. for 5 to 60 minutes.
(4) The soybean extract as described in (1), (2) or (3) above, wherein the unicellular treatment substantially converts the soybean cells into single cells by physical treatment.

Moreover, this invention makes the summary the manufacturing method of the soybean extract of the following (5).
(5) To obtain an extract with more protein (40% or more in solids) than normal broth by hydrothermal treatment of soybean, then physical treatment to single cells, and further extraction with hot water A method for producing a soybean extract characterized by the above.

Moreover, this invention makes the summary the following angiotensin converting enzyme inhibitor of (6).
(6) An angiotensin converting enzyme inhibitor comprising the soybean extract of any one of (1) to (4) above.

Further, the gist of the present invention is the following composition (7).
(7) A composition in a form selected from the group consisting of foods and drinks, feeds, pharmaceuticals, cosmetics, and culture media, containing the soybean extract of any one of (1) to (4) as an active ingredient.

  INDUSTRIAL APPLICABILITY The present invention can provide a component contained in soybean, particularly a component containing an intercellular binding substance, a method for producing the same, and foods, foods, feeds, pharmaceuticals, cosmetics, and culture media using the same.

  As the soybean used in the present invention, a soybean with seed coat may be used as it is, but it is also preferable to use a peeled soybean in order to facilitate unicellularization after hydrothermal treatment. The seed coat can be removed by known physical treatment methods.

  Hot water treatment is required before unicellular treatment. With only the soaking treatment that is usually performed in the production of tofu and soymilk, cells are easily destroyed by a grinding treatment or the like, and target components are not extracted. Immersion treatment may be performed before the hot water treatment, and subsequent single cellization becomes easy.

  The hot water treatment before unicellularization is pressurized according to the water temperature that requires soybeans, and is 100 to 150 ° C., preferably 115 to 125 ° C., at least 2 times the volume of soybeans, preferably 5 times the volume of hot water. And dipping for 5 minutes to 60 minutes, preferably 10 minutes to 20 minutes, while maintaining the temperature of water. You may carry out several times as needed.

  By hydrothermal treatment before unicellularization, the components in soybean are dissolved. Specifically, saccharides such as protein, sucrose, raffinose, and stachyose are obtained. Such an extract is used as a by-product in a lactic acid bacteria fermentation material or the like by a known method. Further, it can be purified by known methods such as acid precipitation, acid alcohol washing, activated carbon adsorption, ion exchange resin, UF membrane and the like.

  The unicellularization referred to in the present invention is preferably that all cells in soybean are completely unicellular, but some of the cells are in contact with other cells via an intercellular adhesive substance. Unicellularization is also included.

  The unicellular treatment referred to in the present invention is performed by physically treating the cells so that the cells are not destroyed after the hydrothermal treatment. For example, it can be processed by a crushing method such as a coffee mill, a food processor, or a mass collider, and other processing methods such as crushing, stirring, mixing, and ultrasonic irradiation with various devices are also possible. After the treatment, the soybean odor can be greatly reduced by separating the moisture and the solid content by filtration or the like. When pulverizing in a dry state without performing hot water treatment, the cells are destroyed, oil and the like flow out of the cells, become emulsion during subsequent hot water extraction, and cannot be separated. The water used for pulverization is required to the extent that cells are not destroyed, and preferably 50% or more.

  The hot water extraction after the unicellular treatment is pressurized according to the water temperature that requires soybean, and is 100 to 150 ° C., preferably 115 to 125 ° C., and has a heat of 2 or more times, preferably 5 or more times that of soybean. It is carried out by immersing in water while maintaining the temperature of water for 5 minutes to 60 minutes, preferably 10 minutes to 20 minutes. After the treatment, the soy extract is obtained by separating water and solids by filtration or the like. After fractionation, the extract can be further collected by washing with water.

  Ordinary soybean broth has a protein content of about 30% in the solid content, whereas the component of the present invention is 40% or more protein. That is, by extracting soybeans with hot water, mixing them into single cells, and further extracting with hot water, a component having more protein (40% or more in solids) than ordinary boiling juice can be obtained.

  The molecular weight of the protein of this component contains a higher molecular fraction (molecular weight of about 40,000 to 130,000) than ordinary soybean broth. Moreover, since there is very little blue smell (soybean odor) peculiar to soybean and it contains a lot of intercellular binding substances, it can be widely used for various uses such as foods and drinks, feeds, pharmaceuticals, cosmetics, and lactic acid bacteria culture media.

For example, lactic acid bacteria can be grown with the soybean extract of the present invention by the following method, and a protein-bacteria complex can be easily obtained.
Sterilize the aqueous solution of this component and aseptically inoculate the pre-cultured lactic acid bacteria. Here, the lactic acid bacteria have a function of decomposing general sugars to generate lactic acid, and examples thereof include Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus lacteis, Lactobacillus plantarum, and the like. However, the present invention is not limited to this, and various lactic acid bacteria such as cocci and bacilli can be used. The characteristic of the growth of lactic acid bacteria according to the present invention is that lactic acid bacteria are precipitated together with the polymer protein as the culture progresses, so that lactic acid bacteria can be easily obtained.
The obtained lactic acid bacterial cells are pre-frozen and then subjected to freeze-drying to obtain powdered lactic acid bacterial cells.

When the aqueous solution of the soybean extract of the present invention was decomposed with pepsin and trypsin, that is, the angiotensin converting enzyme inhibitory activity (ACE inhibitory activity) in the state where the soybean extract of the present invention was digested and absorbed, High ACE inhibitory activity was confirmed.
The soybean extract of the present invention can also be used as an ACE converting enzyme inhibitor. For this reason, the effect of maintaining normal blood pressure is expected even when used as beverages, soups, other foods and drinks, and pharmaceuticals.

Extract Extraction 500 ml of water was added to 100 g of soybean, hydrothermal treatment was performed at 121 ° C. for 15 minutes, and after filtration, soybean broth 1 was obtained. The residue, soybean, was peeled, 500 ml of water was added, hydrothermal treatment was performed at 121 ° C. for 15 minutes, and after filtration, soybean broth 2 was obtained. The residue was pasted with a coffee mill, 500 ml of water was added, hydrothermal treatment was performed at 121 ° C. for 15 minutes, and after filtration, washed with water to obtain the extract. Table 1 shows analytical values of the soy broth 1, soy broth 2, and extract obtained. Moreover, about the protein, molecular weight analysis was performed by HPLC (FIG. 1, FIG. 2). The HPLC conditions in the molecular weight analysis are as follows.
HPLC conditions Column TOSO-G3000SW XL 7.8 × 300mm
Flow rate 1 nl / min
Eluent 0.1M phosphate buffer 0.2M NaCl, pH 7.0
Detector A280mm

  It was found that the soybean extract of the present invention contains a large amount of protein in the extract (solid content of 40% or more) and has very little soybean odor. Furthermore, the protein in the extract was shown to contain a fraction with a molecular weight of about 40,000 to 130,000.

Lactic acid bacteria fermentation of the extract The soybean soup and the soybean extract obtained in Example 1 were freeze-dried to prepare a 1% aqueous solution of the dried product, and Lactobacillus plantarum (5.0 × 10 ⁶ ) was added to 3 ml of the solution. cells) was added and cultured at 30 ° C. for 48 hours, and the viable cell count of each cell suspension was measured by the colony counting method. The suspension was filtered, white precipitate was collected, and the production number was measured in the same manner. The results are shown in Table 2. The protein molecular weight of the precipitate in the column of white precipitation in Table 2 is shown in FIG. 2 and FIG. 3 (HPLC conditions: the same as in Example), and the result of SDS-PAGE is shown in FIG. Further, the turbidity measurement results are shown in Table 3 and FIG.
It was shown that the protein molecular weight of the precipitate consisting of a complex of lactic acid bacteria and protein contains a fraction of 40,000 to 130,000.

Angiotensin converting enzyme inhibitory activity of the extract The inhibitory activity of angiotensin converting enzyme (ACE), which catalyzes the conversion of angiotensin I to angiotensin II, was examined.
It is well known that angiotensin converting enzyme inhibitors are useful for the treatment of essential hypertension. Angiotensin converting enzyme is an enzyme having an important role in the renin-angiotensin system. The enzyme renin secreted from the kidney acts on angiotensin nogen in the blood to produce angiotensin I having weak pressor activity. Angiotensin converting enzyme acts on this to produce angiotensin II, which is the most potent pressor hormone in the body. Angiotensin II, as a direct action, contracts peripheral capillaries and stimulates sympathetic nerves and adrenal glands, and increases blood pressure to promote catecholamine release. Thus, the mechanism of angiotensin converting enzyme is clarified, and its effectiveness in treating hypertension has been confirmed by inhibiting the activity of angiotensin converting enzyme.
Compared to synthetic inhibitors, angiotensin-converting enzyme inhibitors in food ingredients taken daily have no side effects and long-term dietary experience has confirmed their safety. Expected to keep normal.

The soybean broth and soybean extract obtained in Example 1 were lyophilized, and 0.5 ml of 10% solution of the dried product was added to pepsin (Sigma, porcine stomach mucosa, 0.2% with 0.01 N HCl). 0.5 ml of the prepared one) was added and treated at 36 ° C. for 15 hours, followed by centrifugation, and 2.5 ml of 0.1 M Tris buffer (containing 20 mM CaCl ₂ , pH 7.0) was added to the supernatant. Subsequently, 0.5 ml of trypsin (derived from porcine pancreas, adjusted to 0.2% with 0.1 M Tris buffer (containing 20 mM CaCl ₂ , pH 7.0)) was added to 0.5 ml of this solution. After centrifuging at 36 ° C. for 15 hours, centrifugation was performed, and the supernatant was diluted with 2.5 ml of 0.1 M borate buffer (containing 0.3 M NaCl, pH 8.3), and ACE inhibitory activity was measured. did. The measurement results of the ACE inhibitory activity of the soybean extract enzyme-treated solution (pepsin-trypsin digest solution) in the present invention are shown in Table 4 below. This result shows that the soybean extract in the present invention is changed to one having an ACE inhibitory activity in the body by pepsin-trypsin enzyme treatment or when eaten.

The ACE inhibitory activity was measured according to a conventional method, that is, according to a modified method of Cushman et al. (DWCushman et.al. Biochem. Pharamacol., 20,1637), and an angiotensin I converting enzyme (manufactured by Wako Pure Chemical Industries, Ltd., From bovine lung).
0.25 ml of the reaction solution containing the borate buffer solution and 125 μU ACE was added to 0.05 ml of the soybean extract enzyme-treated solution in advance at 37 ° C. for 10 minutes, and 5 mM Hypril-L-histidyl-L-leucine was used as a substrate. The reaction was started by adding 0.25 ml of a substrate solution dissolved in the same borate buffer, and after reacting at 37 ° C. for 30 minutes, 2 ml of 0.1 M NaOH was added to the reaction product to stop the reaction, and then methanol was added. 0.1 ml of 0.2% ortho-phthalaldehyde dissolved in was added and allowed to stand at room temperature for 15 minutes. Thereafter, 0.4 ml of 1.5 M phosphoric acid solution was added, and the fluorescence intensity of the obtained solution was measured using a fluorometer (Shimadzu RF-1500) at an excitation wavelength of 360 nm and a fluorescence wavelength of 480 nm. From the above, the ACE inhibition rate was calculated.
ACE inhibition rate (%) [1- (S-Bs) / (C-Bc)] × 100

S: Fluorescence intensity when ACE and ACE inhibitor sample are added C: Fluorescence intensity when ACE is added and ACE inhibitor sample is not added Bc: Fluorescence intensity when ACE and ACE inhibitor sample is not added Bs : Fluorescence intensity when ACE inhibitor is added without adding ACE

The soybean extract of the present invention can produce lactic acid bacteria cells at a lower cost than when a conventional milk-derived culture solution is used as a material for lactic acid bacteria fermented foods. If a flavor, a sweetener, etc. are added appropriately, it can be used as a food such as a lactic acid fermented beverage. Also, lactic acid bacteria powder can be easily produced by collecting and drying the precipitate.
Further, since the extract itself has little soybean odor, it can be directly used as a food as it is by adding flavors, sweeteners and the like as appropriate. Since the molecular weight is smaller than that of general soy protein foods such as soy milk, it is advantageous in terms of digestion and absorption.
An angiotensin converting enzyme inhibitor can be obtained from soybean, which is a food that has been proven to be safe from a long dietary experience.
It is possible to provide foods, feeds, pharmaceuticals, cosmetics, and culture media that can be expected to have the physiological functions of the components, in which the components containing the intercellular binder contained in the soybean of the present invention are blended.

Protein molecular weight (HPLC) of soybean soup 1, soybean soup 2, and extract of Example 1 Relationship between detection time and molecular weight of Example 1 and Example 2 (HPLC) Protein molecular weight (HPLC) of the lactic acid bacteria fermentation precipitate of the extract of Example 2 Developed photograph of fermentation extract of lactic acid bacteria of Example 2 on SDS-PAGE (stained with 5-20% gradient gel Coomassie Brilliant Blue) Bar graph of turbidity measurement results shown in Table 3

Claims (7)

  Soy extract obtained by hot water extraction of soybeans that have been treated with hot water and single cells.   The soybean extract according to claim 1, wherein the peeled soybean is used.   The soybean extract according to claim 1 or 2, wherein the hydrothermal treatment is to cook soybeans with hot water at 100 to 150 ° C for 5 to 60 minutes.   4. The soybean extract according to claim 1, 2, or 3, wherein the unicellular treatment is a method in which soybean cells are substantially unicellularized by physical treatment.   It is characterized by obtaining an extract with more protein (40% or more in solids) than ordinary boiled juice by hydrothermal treatment of soybeans, then physical treatment to single cells, and hot water extraction. To produce soybean extract.   The angiotensin converting enzyme inhibitor which consists of a soybean extract in any one of Claims 1-4. A composition in a form selected from the group consisting of foods and drinks, feeds, pharmaceuticals, cosmetics, and culture media, comprising the soybean extract of any one of claims 1 to 5 as an active ingredient.