KR20200132329A - Method for producing a fermented soybean product using Aspergillus oryzae - Google Patents

Abstract

The present invention relates to a production method of a fermented soybean product using Aspergillus oryzae capable of producing a high level of free amino acid. The production method of a fermented soybean product comprises the steps of: immersing beans in water of 20 to 45°C, wherein the beans in the step are immersed until the mass thereof is more than doubled; sterilizing the immersed beans; and inoculating Aspergillus oryzae on the sterilized beans and fermenting the same at 28 to 32°C for 48 hours or more, wherein the fermented beans comprise amylase, protease, and free amino acids.

Description

Method for producing a fermented soybean product using Aspergillus oryzae {Method for producing a fermented soybean product using Aspergillus oryzae}

The present invention relates to a method for producing a fermented soybean product using Aspergillus oryzae .

Fermented soybeans have traditionally been used as a food seasoning in Southeast Asia. Fermented soybeans have antioxidant activity (Hu CC, et al., Antioxidant activity of fermented soybean extract, J Agric Food Cam, 52(2), pp. 5735-5739, 2004), and the therapeutic effect of diabetes (Dae Young Kwon). , et al., Antidiabetic effects of fermented soybean products on type 2 diabetes, Nutrition Research, 30(1), pp. 1-13, 2010), and anticancer effects (Keun-Ok Jung, et al., Longer aging time increases the anticancer and antimetastatic properties of doenjang, Nutrition, 22(5), pp. 539-545, 2006) have been reported.

Accordingly, studies on fermentation strains that have high fermentation efficiency of soybeans and that can produce beneficial ingredients for the body when soybeans are fermented have been continued. In particular, when fermenting soybeans using the Aspergillus oryzae GB-107 strain, it was reported that the protein content was increased compared to the existing strain, the trypsin inhibitor was removed, and the size of the peptide was reduced. Yes (Kee-Jong Hong, et al., Aspergillus oryzae GB-107 Fermentation Improves Nutritional Quality of Food Soybeans and Feed Soybean Meals, Journal of medicinal food, 7 (4), pp. 430-435, 2004).

In the present invention, it is intended to increase the content of ingredients useful for humans through fermentation of soybeans.

Kee-Jong Hong, et al., Aspergillus oryzae GB-107 Fermentation Improves Nutritional Quality of Food Soybeans and Feed Soybean Meals, Journal of medicinal food, 7 (4), pp. 430-435, 2004

One object of the present invention is to provide a method of manufacturing a fermented soybean product using Aspergillus oryzae . Another object of the present invention is to provide a method for producing a soybean fermentation product capable of enhancing the amount of secretion of enzymes and the amount of production of free amino acids. Another object of the present invention is to provide a strain in which the amount of enzyme secretion and the amount of free amino acid production are increased.

1. The step of immersing the beans in water of 20 ℃ to 45 ℃, soaking until the mass of the beans is more than twice, the step of; Sterilizing the soaked beans; And inoculating Aspergillus oryzae in sterilized beans and fermenting at 28° C. to 32° C. for 48 hours or more and less than 84 hours, and Aspergillus oryzae is Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241. , The fermented soybean includes amylase, protease, and free amino acid, a method for producing a fermented soybean product.

2. According to item 1, drying the fermented soybeans at 35°C to 45°C so that the moisture content of the soybeans is 10% or less; And deodorizing the dried fermented soybeans.

3. The method according to item 1 or 2, wherein In the fermentation step, Aspergillus oryzae KFCC 11827 is fermented at 30° C. for 72 hours, Aspergillus oryzae KCCM 12698, and Aspergillus oryzae KCCM 60241 are fermented at 26° C. for 72 hours.

4. The method according to item 1 or 2, wherein Aspergillus oryzae is inoculated at 0.15% (w/w) based on the mass of soaked soybeans in the fermentation step.

5. The method according to item 1 or 2, wherein the immersion step is immersed in water at 40° C. for 4 hours.

6. The method according to item 2, wherein the sterilization step is sterilized at 121°C for 40 minutes, the drying step is dried at 40°C for 21 hours, and the deodorizing step is deodorized at 1 Torr±5 for 4 hours.

7. As a strain having the ability to secrete amylase, protease, and free amino acid when fermenting soybeans, the strain is Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241.

8. A composition for fermenting soybeans comprising the strain of item 7.

Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241 according to the present invention secrete enzymes including amylase and protease at a high level compared to conventional strains. In addition, when the soybean is fermented with the strain according to the present invention, a high level of free amino acids can be produced.

1 is a graph showing changes in the weight of beans according to immersion temperature and time.
2 is a photograph showing the color change of beans according to the sterilization time.
3 is a graph showing the contents of amylase, protease, and free amino acid of fermented soybeans prepared in Preparation Examples 1 to 3 according to an embodiment of the present invention, wherein the amylase and protease vertical axis are unit/g, and the free amino acid content The vertical axis is mg/100g.
FIG. 4 is a graph showing the contents of amylase, protease, and free amino acid of fermented soybeans prepared in Preparation Examples 4 to 9 according to an embodiment of the present invention, wherein the amylase and protease vertical axis are unit/g, and the free amino acid content The vertical axis is mg/100g.
5 is a graph showing the contents of amylase, protease, and free amino acid of the prepared soybean fermented product when fermenting soybeans using various Hwangguk bacteria of the present invention, where the amylase, protease vertical axis is unit/g, and the free amino acid content The vertical axis is mg/100g.
6 is a graph showing the contents of amylase, protease, and free amino acid of fermented soybeans prepared in Preparation Examples 15 to 20 according to an embodiment of the present invention, where the amylase and protease vertical axis are unit/g, and the free amino acid content The vertical axis is mg/100g.
7 is a graph showing the change in moisture content according to the drying temperature and time of fermented soybeans.
8 is a graph showing a change in the strength of fermented odor over time when the fermented soybean is deodorized using a freeze dryer.

The invention will now be more fully described below with reference to the accompanying drawings, but only some, but not all, embodiments of the invention are illustrated. Indeed, these inventions can be embodied in many different forms and should not be construed as being limited to the embodiments presented herein. The singular forms used in this specification and the appended claims include plural objects unless clearly indicated otherwise.

The present invention confirmed that the content of amylase, protease, and free amino acid significantly increased in the soybean fermented product fermented using the Aspergillus oryzae strain or Bacillus subtilis strain compared to the case of using the conventional strain (FIGS. 3 to 6 ). .

Beans are the fruit of the annual plant, and beans in the present invention may be used as soybeans, white tae, seoritae, lentils, black beans, chickpeas, kidney beans, lima beans, red beans, red beans, gul red beans, red kidney beans, or migraine, but limited thereto. It does not become.

Free amino acids exhibit sweet, bitter, sour, salty, and umami taste and are used not only as seasonings, but also act as neurotransmitters (Stanislav Reinis, et al., The chemistry of behavior, pp. 355-380, Springer), muscles ( J Bergstroem, P Fuerst, et al., Intracelluar free amino acid concentration in human muscle tissue, Journal of Applied Physiology, 36(6), pp. 693-697, 1974), liver (Wayne L. Ryan, et al., Free amino acids of human foetal and adult liver, Nature, 212, pp. 292-293, 1966), and various tissues including the brain (David S. Dunlop, et al., The presence of free D-aspartic acid in rodents and man, Biochemical and Biophysical Research Communications, 141(1), pp. 27-32, 1986). In addition, the intake of free amino acids can improve muscle mass, strength, and function or slow degeneration (Elisabet borsheim, et al., Effect of amino acid supplementation on muscle mass, strength and physical function in elderly, Clinical Nutrition, 27( 2), pp. 189-195, 2008), the intake of free amino acids can reduce mental fatigue during exercise, increase cognitive performance, and improve motor performance (E. Blomstrand, Amino acids and central fatigue, Amino Acids, 20(1), pp. 25-34, 2001).

Amylase is an enzyme that hydrolyzes starch into sugar and is present in the saliva of humans and other mammals. Proteases, peptidases, or proteinases are enzymes that hydrolyze the peptide bonds of proteins. When a food containing an active amylase or protease is consumed, starch or protein is decomposed into smaller units, so that the digestion of the food is accelerated and the absorption of nutrients is facilitated.

Negotiated fermentation refers to the process of generating energy from carbohydrates in the absence of oxygen, but the term fermentation used in the present invention is a metabolic process that chemically changes the organic substance substrate through the action of enzymes or through the action of microorganisms. It means a process that involves change.

The soybean fermented product manufactured according to the present invention may be prepared in the form of, for example, beverage, gum, tea, powder, granules, tablets, capsules, confectionery, rice cakes, bread, etc., but is not limited thereto. The soybean fermentation product according to the present invention may contain fermented soybeans in a proportion of 0.1% to 100% of the total weight.

The soybean fermented product of the present invention may contain ordinary food additives, and whether it is suitable as a food additive is determined according to the general rules and general test methods of food additives approved by the Food and Drug Administration, unless otherwise specified. It is judged according to standards and standards.

Examples of the items listed in the food additive process include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as reduced pigment, licorice extract, crystalline cellulose, high color pigment, and guar gum; It includes, but is not limited to, mixed preparations such as a sodium L-glutamate preparation, an alkali additive for noodles, a preservative preparation, and a tar color preparation.

For example, the fermented soybean product in the form of a tablet granulates a mixture of the fermented soybean of the present invention with an excipient, a binder, a disintegrant and other additives in a conventional manner, and then compression-molded by adding a lubricant or the like, The mixture can be directly compression molded. In addition, the fermented soybean product in the form of a tablet may contain a mating agent or the like as necessary.

Among the capsule-type soybean fermentation products, the hard capsule can be prepared by filling a mixture of the fermented soybean product of the present invention with an excipient, etc. into a conventional hard capsule, and the soft capsule is the fermented soybean product of the present invention as an excipient, etc. It can be prepared by filling a mixture mixed with the additive of the capsule base such as gelatin. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

The ring-shaped soybean fermentation product can be prepared by molding a mixture of the fermented soybean product of the present invention with excipients, binders, disintegrants, etc. by conventionally known methods, and can be coated with white sugar or other coating agents if necessary. Alternatively, the surface may be coated with a material such as starch or talc.

The soybean fermentation product in the form of granules can be prepared in granular form by a mixture of the fermented soybean product of the present invention, excipients, binders, and disintegrants, etc., and contains flavoring agents, flavoring agents, etc. as needed. can do.

For example, the soybean fermentation product of the present invention may include one or more additional ingredients for improving flavor, such as apple powder, sweet pumpkin powder, soybean powder, or roasted soybean powder together with the soybean fermentation product. It was found that mixing apple powder or sweet pumpkin powder with fermented soybeans can increase taste preference and solubility, and lower fermentation odor.

The method of manufacturing a soybean fermentation product according to the present invention includes the step of immersing soybeans in water, sterilizing it, and fermenting it with Aspergillus oryzae or Bacillus subtilis . When the soybean fermentation product is manufactured by the method according to the present invention, the content of the free amino acid as well as the content of enzymes including amylase and protease is increased compared to the prior art. The method according to the invention may further comprise drying the fermented beans and/or deodorizing the dried fermented beans.

Hereinafter, the process will be described in detail.

The step of immersing the beans is a process for softening the beans, and may be performed by immersing the beans in purified water 5 to 10 times the weight of the beans. The temperature of the purified water may be performed at 4 ℃ to 40 ℃. If the temperature of purified water is lower than 4℃, the time to soften the beans becomes longer, resulting in lower productivity. If the temperature is higher than 40℃, the difference in the degree of softening and moisture content of the beans increases according to the change of soaking time. Problems arise in manufacturing a quality product.

When the mass of the soaked beans becomes about 2.2 times the original mass of the beans, the rate of increase decreases significantly, so it is preferable to immerse them until about 2.2 times the mass of the beans before immersion. The immersion time may be 1 hour to 20 hours, but it may be easily changed and used by a person skilled in the art according to the temperature of the purified water used in the immersion process. For example, about 8 hours when immersed at normal pressure, 25°C; About 6 hours when immersed at normal pressure, 30°C; In the case of immersion at normal pressure and 40°C, soybeans having the desired physical properties can be prepared by immersing for about 4 hours.

The soaked beans are subjected to a sterilization step. The sterilization method can be performed according to a method known in the art. For example, the dry heat method of removing bacteria by maintaining a high temperature in a dry state, the moist heat method to remove bacteria by maintaining a high temperature in a humid state, a filtration sterilization method that filters out microorganisms using a fine filter paper, and electromagnetic waves are used. Radiation sterilization method in which bacteria are removed by irradiation, and gas sterilization method in which microorganisms are killed by spraying a specific compound can be used. The dry heat method includes a flame sterilization method that kills microorganisms by directly contacting an object to be sterilized with a flame, and a dry heat sterilization method that oxidizes or carbonizes the microorganisms using a dry heat sterilizer, and the wet heat method is a boiled boiled sterilization method in water at 100°C, from 62°C. There are a pasteurization method of sterilization by heating at 65°C or 75°C, a high-pressure steam sterilization method of sterilization under high pressure conditions at 121°C, and an intermittent sterilization method by heating intermittently several times at 100°C. The radiation sterilization method sterilizes microorganisms by irradiating ultraviolet rays, X-rays, neutrons or cathode rays, and the gas sterilization method is a method of sterilizing microorganisms by adding formaldehyde, ethylene oxide, or beta-propiolactone.

A person skilled in the art may perform a sterilization method depending on the object to be sterilized, and in the present invention, sterilization was performed by high-pressure steam sterilization using an autoclave, but is not limited thereto. In particular, in one embodiment of the present invention, sterilization is performed at 121° C. 2 atm for 30 minutes to 50 minutes, and in another embodiment at 121° C. 2 atm for 40 minutes. Depending on the sterilization conditions, differences in color, texture, and organoleptic properties of the pretreated beans occur, and if sterilized for less than 30 minutes, the beans are hard and may cause problems in performing the subsequent process, and sterilize for more than 50 minutes. In this case, the beans become too soft and cause a hot odor, making them unsuitable for use as food.

The fermentation process is a step of fermenting soybeans with Hwangguk-gyun ( Aspergillus oryzae ), or Bacillus subtilis . The material contained in the soybean is transformed into a form suitable for consumption through the fermentation process, and in particular, it is fermented using Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241 according to the present invention, or Bacillus subtilis KCCM 10839 The resultant fermented by fermentation with high levels of amylase, protease, and/or free amino acids. The amylase, protease, and/or free amino acid contained in the soybean fermentation product prepared according to the present invention exhibits a higher level compared to the case of fermentation by a conventionally known strain, and this difference is due to the specificity of the strain itself and the present invention. This is due to the optimized conditions of the process accordingly.

Fermentation in the case of Aspergillus oryzae KFCC 11827, in one embodiment of the present invention, 20 hours to 84 hours at 28 ℃ to 32 ℃, 20 to 84 hours at 30 ℃ in another embodiment of the present invention, another embodiment of the present invention In an example, it is fermented at 30° C. for 48 hours to 72 hours, and in another embodiment of the present invention at 30° C. for 72 hours. In particular, the present inventors found that the content of amylase, protease, and free amino acids in the soybean fermented product increased significantly between 25 and 48 hours when fermented at 30°C, and nonetheless, the degree of fermentation did not increase significantly. Was confirmed. In addition, Aspergillus oryzae KCCM 12698, KCCM 60241 or Aspergillus oryzae roneun is preferred when it fermenting in 26 ℃ for 72 hours considering the soybean fermentation amylase, protease, and levels of free amino acids in water. When fermenting with Bacillus subtilis KCCM 10839, it is preferable to ferment at 40° C. for 72 hours.

In one embodiment of the present invention Aspergillus oryzae is 0.01% (w/w) to 1% (w/w) based on the mass of soaked beans, in another embodiment of the present invention 0.10% to 0.5% (w/w ), In another embodiment of the present invention, about 0.15% (w/w) is inoculated, and Bacillus subtilis is 0.01% (w/w) to 1% (w/w), based on the mass of soaked beans. In another embodiment of the present invention, inoculation is performed at 0.05% to 0.2% (w/w), and in another embodiment of the present invention, about 0.10% (w/w). The amount of inoculated strain is 0.01% (w/w) If it is less than 1% (w/w), fermentation efficiency decreases significantly, and if it exceeds 1% (w/w), preference is lowered due to aroma generated by microorganisms.

In one embodiment of the present invention, soybeans may be pulverized and used before fermentation. When the beans are pulverized, fermentation efficiency can be improved because the cross-sectional area that can be in contact with microorganisms is widened. In particular, in the case of fermentation with Bacillus subtilis KCCM 10839, fermentation efficiency is remarkably increased by crushing and fermenting sterilized soybeans.

The method according to the present invention may further include drying the fermented soybeans and/or deodorizing the dried fermented soybeans. Drying the fermented soybeans may make it easier to transform or manipulate into a desired form of food. In addition, it is possible to increase the preference of fermented soybean products by reducing the smell generated during fermentation by passing through the deodorization process.

In one embodiment of the present invention, the drying step is performed at 20°C to 50°C, and in another embodiment of the present invention, 35°C to 45°C. Although it is possible to adjust according to conventional knowledge, it is preferable to dry so that the moisture content of the beans is 10% or less, and the drying time may be adjusted according to the desired moisture content and drying temperature. For example, drying for 30 hours or more at 35°C, 21 hours or more at 40°C, or 18 hours or more at 45°C may reach a desired moisture content, but is not limited thereto.

The deodorization step may be performed according to a method well known in the art. Examples of usable deodorization methods include physical treatment methods including water washing, activated carbon adsorption, and air dilution; A chemical treatment method including an oxidation method including an ozone oxidation method and a chlorine oxidation method, a chemical solution method including an acid/alkali washing method, an ion resin exchange method, a combustion method including a direct combustion method and an indirect combustion method, a neutralizing method, and a masking method; And a liquid-gas treatment method including scrubber deodorization and aeration tank deodorization, and a solid-gas treatment method including soil deodorization, carrier-filled deodorization, and immobilized microorganism deodorization may be used.

The strain used in the method according to the present invention secretes high levels of amylase and protease when fermenting soybeans, and the soybean fermentation product fermented with the above strain contains a high level of free amino acids, and the strain is Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae 60241, or Bacillus subtilis KCCM 10839. In an embodiment of the present invention, the soybean fermented product produced using the strain is at least about 1,800 unit/g or more amylase, 2,000 unit/g or more amylase, 3,000 unit/g or more amylase, 10,000 unit/g or more amylase, or It may comprise 20,000 units/g or more amylase and/or, in one embodiment of the present invention, may comprise 100 unit/g or more protease, 200 unit/g or more protease, or 300 unit/g or more protease, and/or, It may contain 600 mg/100g or more free amino acid, 800 mg/100g or more free amino acid, 1,000 mg/100g or more free amino acid, or 2,000 mg/100g or more free amino acid.

The present invention provides a composition for fermentation of beans comprising Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae 60241, and/or Bacillus subtilis KCCM 10839.

The content of free amino acids is determined by Article 8. General Test Methods of the Food Code / 2. Food Components Test Methods / 2.1 General Components Test Methods / 2.1.3 Nitrogen Compounds / 2.1.3.3 Amino Acids / a. It was measured by referring to the quantification method by an automatic amino acid analyzer, and the amino acid content was calculated using an amino acid standard solution with known HPCL and concentration (Food Code, Food Safety Nara, 2019.01.31.2019-7).

In addition, the content of enzyme is determined by using a spectrophotometer in the sample according to the method described in Article 8. General Test Methods / 6. Food Standard Confirmation Test Methods / 6.8 Agricultural Processed Foods / 6.8.3 Enzyme Foods / 6.8.3.2 Protease. The content of the enzyme contained was quantified.

Hereinafter, the present invention will be described with specific examples.

Example

Example 1. Immersion

100 g of soybeans (white tae, soybean) were immersed in water at 4°C, 25°C, 30°C and 40°C for 8 hours, and the weight of the soaked beans was measured every hour. The results are shown in Table 1 and FIG. 1 below.

division 0 hours 1 hours 2 hours 3 hours 4 hours 5 hours 6 hours 7 hours 8 hours cold storage 100 129.2 139.9 148.9 156.8 163.6 170.4 176.5 181.9 Room temperature (25℃) 100 144.8 166.6 186.2 199.3 207.25 215.2 218.8 220.2 30℃ 100 155.5 179 198.3 210 215.45 220.9 222.5 224.3 40℃ 100 171.4 202.8 216.7 222 223.15 224.3 225.5 226

The higher the temperature, the faster the bean weight gained. When the initial weight reached about 2.2 times, the texture including hardness, elasticity, guminess, chewiness, and cohesiveness appeared similar, and the moisture content was 38% to 40% (w/w), indicating no significant difference. .

According to the above results, beans were immersed at 40° C. for 4 hours below and then used.

Example 2. Sterilization

The soaked beans according to Example 1 were sterilized in an autoclave at 121° C. for 30 minutes, 40 minutes and 50 minutes, respectively. The number of bacteria before sterilization was 4.16±0.16 (log ₁₀ CFU/g), and no bacteria were detected after sterilization.

Color, texture, and sensory properties before and after sterilization were evaluated according to each time, and the results are shown in Table 2 below, and pictures taken of each are shown in FIG. 2.

Heat treatment condition After immersion and before heat treatment 121℃ for 30 minutes 121℃ 40min 121℃ for 50 minutes color Bright white phase Light brown Brown Dark brown Texture stiffness Slightly hard Soft Very soft Sensuality Has fishy smell good good Occurrence of fever, etc.

In particular, it was confirmed that the fishy smell of beans was removed before and after sterilization. Raw soybeans contain lipoxygenase, and lipoxygenase breaks down the fat components of soybeans to generate compounds that cause fishy odors such as n-hexanal. During the sterilization process, the enzyme activity of lipoxygenase is As it was removed, the fishy smell also appeared to be reduced.

Meanwhile, sterilization at 121° C. for 40 minutes was found to be the most excellent in terms of color, texture, and sensory properties. Hereinafter, the beans were sterilized at 121°C for 40 minutes and then used.

Example 3. Fermentation

3.1 Fermentation by Hwangguk-gyun

According to Examples 1 and 2, 1.5 g of Hwangguk bacteria ( Aspergillus oryzae KFCC 11827) was inoculated into 1,000 g of soybeans that were immersed in water at 40° C. for 4 hours and sterilized at 121° C. for 40 minutes in an autoclave, Fermentation was performed at 28° C., 30° C., and 32° C. for 20 hours each (Preparation Examples 1 to 3, respectively). The fermented soybeans were dried at 40° C. for 20 to 22 hours, pulverized to have an average particle size of 90 μm, and then the amylase, protease, and free amino acid contents were measured, and the fermented odor was examined. The results are shown in Table 3 and It is shown in Figure 3.

division Before fermentation Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Original 100% beans Whether to escape Do not escape Immersion 40℃ 4h Sterilization 121℃ 40m Whether the beans are crushed before fermentation Never zymogen No fermentation Hwang Guk-gyun Starter No fermentation Hwangguk-gyun 1.5g/1kg ( Aspergillus oryzae KFCC 11827) Starter content 0 0.15% (w/w) based on dipped beans Fermentation No fermentation 28℃ 20h 30℃ 20h 32℃ 20h dry 40℃ 20h to 22h smash 160 mesh pin milling (average particle size 90μm) Amylase (unit/g) 0 1,832 2,529 1,953 Protease (unit/g) 0 105 122 103 Free amino acid content (mg/100g) 313 650 703 683 Fermented odor strength No fermentation odor - + -

As a result, the amylase content contained in the soybean powder fermented at 28° C., 30° C., and 32° C. was 1,832 unit/g, 2,529 unit/g, and 1,953 unit/g, respectively, and the protease content was 105 unit/g, respectively, 122 unit/g, and 103 unit/g, and the free amino acid content was 650 mg/100g, 703 mg/100g, and 683 mg/100g. On the other hand, in the case of fermented odor, fermented at 30°C was slightly higher than when fermented at 28°C or 32°C. In conclusion, when Aspergillus oryzae KFCC 11827 was used, fermentation at 30℃ had the highest content of amylase, protease, and free amino acids in fermented soybean powder.

Accordingly, changes in the contents of amylase, protease, and free amino acids and the strength of fermentation odor were confirmed according to the fermentation time at 30° C. and whether or not crushed, and the results are shown in Table 4 and FIG.

division Manufacturing Example 4 Manufacturing Example 5 Manufacturing Example 6 Manufacturing Example 7 Manufacturing Example 8 Manufacturing Example 9 Original 100% beans Whether to escape Do not escape Immersion 40℃ 4 hours Sterilization 121℃ 40min Whether the beans are crushed before fermentation Never Shred to 1/5 level Never zymogen Hwang Guk-gyun Starter Hwangguk-gyun 1.5g/1kg ( Aspergillus oryzae KFCC 11827) Starter content 0.15% (w/w) based on dipped beans Fermentation 30℃ 25h 30℃ 48h 30℃ 60h 30℃ 72h 30℃ 72h 30℃ 84h dry 40 ℃ 20 to 22 hours smash 160 mesh pin milling (average particle size 90μm) Amylase (unit/g) 3,256 16,324 19,591 22,858 25,928 23,843 Protease (unit/g) 135 257.33 287.92 319 180 305 Free amino acid content (mg/100g) 878 1,686 2,108 2,842 2,929 3,010 Fermented odor strength + ++ ++ ++ ++ ++++ ^*

As a result, the content of amylase, protease, and free amino acid tended to increase as the fermentation time increased at 30° C., but when fermented for 84 hours, it was confirmed that the content of amylase and protease decreased. In addition, the contents of amylase, protease and free amino acids were rapidly increased in Preparation Example 5 (30°C 48h) compared to Preparation Example 4 (30°C 25h). On the other hand, in the case of Preparation Example 8 in which soybeans were crushed before fermentation, the content of amylase and free amino acid were slightly increased, but the protease content was significantly decreased as compared to Preparation Example 7 in which the soybeans were crushed and fermented under the same conditions. Became. In addition, as the fermentation time increased from 72 hours to 84 hours, the fermented odor was greatly increased, which appeared to be unsuitable as a food.

As a result, it was confirmed that fermentation at 30° C. for 72 hours is the most suitable condition.

Further, soaked for 4 hours at 40 ℃ water, the pre-sale received Aspergillus oryzae KCCM 12698, and Aspergillus oryzae KCCM 60241 1.5g seed from the Korea Association in which a 121 ℃ sterilized for 40 minutes at an autoclave, 1,000 g soya did not break each After inoculation, each strain was incubated for 72 hours at 26°C, which is the desired culture temperature. The fermented soybeans were dried at 40° C. for 20 to 22 hours, pulverized to have an average particle size of 90 μm, and then the amylase, protease, and free amino acid contents were measured, and the fermented odor was examined. In order to compare the results, a product manufactured by the method described in Korean Patent Laid-Open Patent No. 10-2007-0009248 was used as a control. The results are shown in Table 5 and FIG. 5.

division KFCC 11827 KCCM 12698 KCCM 60241 Contrast Whether the beans are crushed before fermentation Not shredded Shreds Fermentation 30℃ 72h 26℃ 72h 26℃ 72h 50℃ 60min Amylase 22,858 2,346 2,110 3,997 Protease 319 253 238 10 Free amino acid content
(mg/100g) 2,842 2,265 3,298 556 Fermented odor strength ++ ++ ++ ++++

In terms of the content of amylase, protease and free amino acid, Aspergillus oryzae KFCC 11827 was found to be the best overall, and particularly, it was confirmed that the content of amylase was remarkably high. In addition, compared to the control group, the amount of amylase was 5 times or more, the protease content was 30 times or more, and the free amino acid content was 5 times or more, and the fermented odor was also significantly lower than the control.

3.2 Fermentation by Bacillus Bacillus

Bacillus subtilis KCCM 10839 and 1.5 g of Bacillus subtilis purchased from Yuzo Takahashi were respectively inoculated into 1,000 g of non-shelled soybeans immersed in water at 40° C. for 4 hours and sterilized at 121° C. for 40 minutes in an autoclave. It was fermented for 20 to 84 hours at 43°C, respectively. The fermented soybeans were dried at 40° C. for 20 to 22 hours, pulverized to have an average particle size of 90 μm, and then the contents of amylase, protease, and free amino acid were measured, and the fermented odor was examined, and the results are shown in Table 6, It is shown in Table 7 and Figure 6.

division Before fermentation Manufacturing Example 10 Manufacturing Example 11 Manufacturing Example 12 Manufacturing Example 13 Manufacturing Example 14 Original 100% beans 100% beans 100% beans 37% beans, 37% brown rice, and 26% barley 100% beans 100% beans Whether to escape Do not escape Immersion 40℃ 4 hours Sterilization 121℃ 40min Whether the beans are crushed before fermentation Never zymogen No fermentation Bacillus Bacillus Starter - KCCM 10839 Yuzo Takahashi Yuzo Takahashi KCCM 10839 KCCM 10839 Starter content 0 0.1% based on dipped beans (w/w) Fermentation No fermentation 40℃ 20h 40℃ 20h 40℃ 20h 37℃ 20h 43℃ 20h dry 40 ℃ 20 to 22 hours smash 160 mesh pin milling (average particle size 90μm) Amylase (unit/g) 0 Not detected Not detected 2,443 Not detected Not detected Protease (unit/g) 0 35 60 23 42 53 Free amino acid content (mg/100g) 313 1,032 503 305 832 753 Fermented odor strength No fermentation odor + + + + +

As a result of fermenting soybeans with KCCM 10839 and Yuzo Takahashi (Preparation Examples 10 and 11), amylase was not detected at all. This was determined to be due to the fact that a significant amount of soy carbohydrates are indigestible carbohydrates such as stachyose. Accordingly, brown rice and barley containing a large amount of easily digestible carbohydrates were fermented together (Preparation Example 12), and as a result, it was confirmed that amylase was secreted.

In addition, in the case of KCCM 10839, when fermented at 40°C, the protease content was the lowest, but the free amino acid content was significantly higher than that at 37°C or 43°C. Accordingly, KCCM 10839 was judged to be the most effective at 40°C, and the difference in effect according to fermentation time and crushing before fermentation was confirmed.

division Manufacturing Example 15 Manufacturing Example 16 Manufacturing Example 17 Manufacturing Example 18 Manufacturing Example 19 Manufacturing Example 20 Original 100% beans Whether to escape Do not escape Immersion 40℃ 4 hours Sterilization 121℃ 40min Whether the beans are crushed before fermentation Never Shred to 1/5 level zymogen Bacillus Bacillus Starter manufacturer Bacillus subtilis KCCM 10839 Starter content 0.1% based on dipped beans (w/w) Fermentation 40℃ 25h 40℃ 48h 40℃ 60h 40℃ 72h 40℃ 72h 40℃ 84h dry 40 ℃ 20 to 22 hours smash 160 mesh pin milling (average particle size 90μm) Amylase (unit/g) Not detected Not detected Not detected Not detected 2,589 2,853 Protease (unit/g) 62 95 112 126 200 230 Free amino acid content (mg/100g) 1,990 2,871 3,331 3,791 4,642 4,832 Fermented odor strength + + + +++ +++ ++++*

As a result, in the case of amylase, although no amylase was detected when uncrushed soybeans were used, the contents of amylase, protease and free amino acids tended to increase over time. In the case, it was confirmed that the contents of amylase, protease, and free amino acid significantly increased. In addition, when fermented for 84 hours, it was judged to be unsuitable for use as food because of the strong fermentation odor.

Example 4. Drying

Soaked in water at 40° C. for 4 hours, sterilized at 121° C. for 40 minutes in an autoclave, and inoculated with 1.5 g of Hwangguk bacteria ( Aspergillus oryzae KFCC 11827) in 1,000 g of soybeans that were not shredded and crushed in an autoclave for 72 hours. After fermentation, the moisture content was measured for each drying time while drying at 35°C, 40°C and 45°C for 30 hours (Table 8, FIG. 7).

Drying temperature Drying time(h) Moisture content (%) Drying temperature Drying time(h) Moisture content (%) Drying temperature Drying time(h) Moisture content (%) 35℃ 3 38.5 40℃ 3 38.5 45℃ 3 38.5 6 34.6 6 33.7 6 30.1 9 31.4 9 29.1 9 22.4 12 28.2 12 23.2 12 14.3 15 26 15 18.9 15 10 18 22.2 18 12.3 18 9.5 21 19.1 21 9.5 21 8.5 24 16.3 24 8.8 24 8 27 12.8 27 8.1 27 8 30 9.6 30 7.9 30 7.5

It was confirmed that the arrival time at which the moisture content was 10% or less was 30 hours at 35°C, 21 hours at 40°C, and 18 hours at 45°C.

However, as a result of comparing the activity of α-amylase after drying until it reaches 10% moisture content at each temperature, when the activity before drying is 100%, 95% and 40°C when dried at 35°C for 30 hours When dried for 21 hours at 93%, when dried at 45 ℃ for 18 hours, only 52% of the activity remained. Further, in the case of protase, only 92% of the activity was left when dried at 35°C for 30 hours, 89% when dried at 40°C for 21 hours, and 43% when dried at 45°C for 18 hours. When heat is applied to a protein, it is judged that the activity decreases due to denaturation. Based on these results, it was confirmed that the critical thermal denaturation temperature of the enzymes was 40°C to 45°C. In consideration of work efficiency and degree of denaturation, drying at 40° C. for 21 hours is considered to be most desirable.

Example 5. Deodorization

Soaked in water at 40° C. for 4 hours, sterilized at 121° C. for 40 minutes in an autoclave, and inoculated with 1.5 g of Hwangguk bacteria ( Aspergillus oryzae KFCC 11827) in 1,000 g of soybeans that were not shredded and crushed in an autoclave for 72 hours. After fermentation, the beans dried at 40° C. for 21 hours were not separately frozen using a freeze dryer, and a sample at room temperature was deodorized, and the vacuum degree was 1 Torr±5 for 4 hours. The strength of fermented odor by time was measured for 10 researchers (Table 9, Fig. 8).

Deodorization time 0 1 hours 2 hours 3 hours 4 hours Fermented odor strength 100% 70% 60% 55% 50%

The fermentation odor of fermented soybeans could be significantly reduced through the deodorization process.

Example 6. Formulation

After pulverizing the soybean fermented product prepared according to Example 5, powder:water was blended in a ratio of 1:6 to evaluate the strength, taste preference, and solubility of the fermented odor in 10 people. In order to compare the results, a product manufactured by the method described in Korean Patent Laid-Open Patent No. 10-2007-0009248 was used as a control (Table 10). Taste acceptability is very poor with a full score of 5: 1; Bad: 2 points; Moderate: 3 points; Good: 4 points; Very good: It was surveyed with 5 points.

division Manufacturing Example 21 Manufacturing Example 22 Manufacturing Example 23 Manufacturing Example 24 Manufacturing Example 25 Contrast Hwangguk-gyun fermented powder 100 0 50 50 50 Bacillus fermented powder 0 100 10 10 10 Soybean powder 0 0 20 10 10 Roasted bean powder 0 0 20 10 10 Apple powder 0 0 0 20 0 Sweet pumpkin powder 0 0 0 0 20 Sum 100 100 100 100 100 Amylase (unit/g) 22,858 2,589 11,688 11,688 11,688 3,997 Protease (unit/g) 318 200 179 179 179 10 Free amino acid (mg/100g) 2,842 4,642 1,885 1,885 1,885 556 Fermented odor strength ++ +++ + + + ++++ Taste preference 2.5 2.5 3.5 4 3.8 2.5 Solubility Somewhat lumpy Somewhat lumpy Somewhat lumpy Well loose Well loose Somewhat lumpy

As a result, when the content of the fermented powder was 100% (Preparation Examples 21 and 22), significantly higher levels were observed in amylase, protease, and free amino acids than in the control examples.

In addition, the present inventors blended soybean powder, roasted soybean powder, apple powder, and/or sweet pumpkin powder in order to enhance taste preference. Soybean powder is dried and pulverized after boiling and peeling beans, and roasted soybean powder is powdered by roasting beans in a popping machine. In addition, apple powder was prepared by peeling apples, then sliced and dried at 60°C for 5 days in a hot air dryer. Sweet pumpkin powder was steamed after peeling sweet pumpkins, dried at 50°C in a hot air dryer for 2 days, and then powdered.

When soybean powder, roasted soybean powder, apple powder, and/or sweet pumpkin powder were mixed with the soybean fermented powder, the fermented odor was significantly reduced and taste preference was increased. It was found to dissolve well in water.

For example, for the purposes of claim construction, the claims set forth below should not be construed in any way narrower than their literal language, and therefore exemplary embodiments from the specification should not be read as claims. Accordingly, it should be understood that the invention has been described as an example and is not a limitation on the scope of the claims. Accordingly, the invention is limited only by the following claims. All publications, issued patents, patent applications, books and journal articles cited in this application are each incorporated herein by reference in their entirety.

Korea Microorganism Conservation Center (domestic) KFCC11827 20190411

Claims (8)

Immersing the beans in water of 20°C to 45°C, immersing the beans until the mass of the beans is twice or more;
Sterilizing the soaked beans; And
Inoculating Aspergillus oryzae on sterilized soybeans and fermenting at 28°C to 32°C for 48 hours or more and less than 84 hours,
The Aspergillus oryzae is Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241, and the fermented soybean includes amylase, protease, and free amino acid, a method for producing a fermented soybean product. The method of claim 1, further comprising: drying the fermented beans at 35°C to 45°C so that the moisture content of the beans is 10% or less; And
The method further comprising the step of deodorizing the dried fermented beans. The method according to claim 1 or 2, wherein In the fermentation step, Aspergillus oryzae KFCC 11827 is fermented at 30° C. for 72 hours, Aspergillus oryzae KCCM 12698, and Aspergillus oryzae KCCM 60241 are fermented at 26° C. for 72 hours. The method according to claim 1 or 2, wherein In the fermentation step, Aspergillus oryzae is inoculated at 0.15% (w/w) based on the mass of soaked soybeans. The method according to claim 1 or 2, wherein the immersion step is immersed in water at 40° C. for 4 hours. The method of claim 2, wherein the sterilization step is sterilized at 121°C for 40 minutes, the drying step is dried at 40°C for 21 hours, and the deodorizing step is deodorized at 1 Torr±5 for 4 hours. As a strain having the ability to secrete amylase, protease, and free amino acid when fermenting soybeans, the strain is Aspergillus oryzae KFCC 11827, Aspergillus oryzae KCCM 12698, or Aspergillus oryzae KCCM 60241. A composition for fermenting soybeans comprising the strain of claim 7.

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