KR101815637B1 - Manufacturing method of peptide natural seasoning containing microorganism white soybean - Google Patents

Abstract

A method for preparing a high-fat natural seasoning containing fermented soybean according to the present invention comprises the steps of: washing soybeans at least two times and then washing the soybeans to remove foreign substances; soaking the soybeans from which the foreign substances have been removed at a predetermined temperature and for a predetermined period of time; A step of removing moisture from the soaked soybeans by supporting the soaked soybeans on the sieve for 1 hour, sterilizing the soybeans in an auto-clave at a predetermined temperature, a constant atmospheric pressure and a predetermined time, , Cooling the sterilized soybeans, immersing the cooled soybeans in a Petri dish on a clean bench, and inoculating B. coagulans ( Bacillus coagulans ) into the Petri dish to prepare fermented soybean .
Accordingly, in the fermented soybean produced in the above step, the fermented soybeans produced are mixed with radish, cola bean, mushroom, green onion, maize, lotus root, garlic, shrimp, clam, cod roe and green pepper.

Description

Technical Field [0001] The present invention relates to a method for manufacturing a high-peptide natural seasoning containing fermented soybean,

The present invention relates to a process for preparing high-peptide natural seasoning containing fermented soybean,

More particularly, the present invention relates to a fermented soybean containing fermented soybean which can be applied to various foods by producing a natural seasoning containing fermented soybean to improve the richness and richness of the fermented soybean, Peptide natural seasoning.

Generally, sodium glutamate (MSG) is typically used as a method of improving the taste of food in a seasoning. Since Dr. Ikeda of Japan in 1908 has separated sodium glutamate from kelp, the nucleic acid component (sodium inosinate / sodium guanylate) has been found to be an ingredient that gives the food a rich taste. As the economy develops, consumers are demanding a higher quality of taste for food, and they want to give the food a more "ripened" and "richer" flavor than the original richness. These aged and abundant flavors are expressed in "Kokumi" in Japan, "mouthfulness" in the United States and the United Kingdom, and "deep taste" and "richness" in our country.

Usually, fermented or aged miso or soy sauce is used to give such a rich feeling to foods.

It is presumed that the peptides and amino acids present in the soybean are the major components of the taste. They are produced by proteolytic enzymes of the germs in the manufacturing process and are produced by decomposing the raw wheat or soybean protein under high salt concentration for a long time. Because of the high concentration of sodium chloride, this type of soup is difficult to use universally for food.

In addition, in order to impart a richness to conventional foods, natural extracts such as meat extract, fish and shell extract, and vegetable extract have been widely used. However, since they have an unpleasant odor and bitter taste derived from raw materials, their use amount is limited depending on the use. It can not be granted.

Peptide that gives a rich taste has been tried, but it has been difficult to obtain a desired effect because it has a complicated separation and purification, has a limited use due to its high price, and has an unpleasant taste and odor such as a bitter taste.

Korean Patent Laid-Open No. 10-2006-0003858, which has been conventionally conducted, discloses a method for dissolving wheat gluten enzymatically decomposed seasoning in water to provide a sugar peptide and a peptide having a rich taste-imparting function, separating the obtained aqueous solution into fractions And then fractionating the resulting fraction having a molecular weight of 1,000 or more is further fractionated by gel filtration, reverse phase HPLC or the like (Patent Document 1).

As a method for producing a seasoning using a mixture of defatted soybean and wheat or wheat gluten as a raw material in the conventional seasoning manufacturing method, Korean Patent Laid-open Publication No. 10-2005-0116784 discloses a method for producing a seasoning comprising 10 to 30 parts by weight of a starch raw material, Wherein the proteinaceous yeast is hydrolyzed at 52 to 60 DEG C for 18 to 30 hours in the presence of salt or in the presence of low salt. In this method, defatted soybean is used as a raw material for protein, and wheat ground material is used as a starch raw material. The defatted soybean content is mostly occupied by 70 to 90 parts by weight, It is used as a raw material and is not used for imparting a rich taste but used for pollution prevention (Patent Document 2).

Furthermore, as a method for producing seasonings using wheat gluten and soybean, Korean Patent Registration No. 10-0623940 discloses a method of producing 100 ~ 60% of raw materials consisting of 25 ~ 100% of gluten and 75 ~ To 40% of the raw material, and then the resulting yeast is placed in a saline solution having a concentration of 1.35 to 1.65 times the concentration of the raw material in a concentration of 7 to 24%, and the mixture is brewed at a temperature of 10 ° C for 2 to 3 months, And then brewing for 1 to 2 months at 10 ° C and then for 1 to 2 months at a temperature of 20 ° C. As a result, although wheat gluten and soybeans were used, (Patent Document 3).

Korean Patent Publication No. 10-2006-0003858 (2006.01.11). Korean Patent Publication No. 10-2005-0116784 (December 13, 2005). Korean Patent No. 10-0623940 (2006.09.07).

It is an object of the present invention to provide a fermented soybean containing fermented soybean which contains a large amount of peptides by using a natural raw material containing fermented soybeans to provide a fermented soybean which can be used for general purposes as well as enhancing and sustaining the basic taste, And to provide a method for manufacturing a high-peptide natural seasoning.

It is still another object of the present invention to provide a method for producing a high-fat natural seasoning containing fermented soybean which improves the overall efficiency of natural seasoning, thereby maximizing the satisfaction and reliability of utilization for those who cook or cook the food. .

In order to solve the above-mentioned problems, the present invention provides a method for producing a high-peptide natural seasoning containing fermented soybean, comprising the steps of: washing soybeans more than two times to remove foreign substances; A step of immersing the soaked soybeans in water for a predetermined period of time; a step of supporting the soaked soybeans on the sieve for 1 hour to remove moisture; and a step of putting the soybeans from which moisture has been removed in the autoclave, Incubating the sterilized soybeans in a glass container in a clean bench after the cooled soybeans are cooled, and inoculating B. coagulans ( Bacillus coagulans ) into the glass container to prepare fermented soybeans .

The sterilization may be performed at a temperature of 110 to 130 ° C, a pressure of 1 to 2 atmospheres, a time of 20 minutes to 10 minutes, It takes 40 minutes.

Accordingly, the present invention provides a fermented soybean produced by mixing fermented soybeans with radish, cola, moss, mackerel, marjoram, lotus root, garlic, shrimp, clam, Weight, 12.9% by weight, 12.9% by weight of cola, 12.9% by weight of mussel, 6.15% by weight of rapeseed, 6.15% by weight of shrimp, 6.25% by weight of lime root, 1.26% by weight of garlic, 1.26% by weight of shrimp, 6.15% by weight and 2.13% by weight of green pepper.

The present invention uses a natural raw material containing fermented soybeans to contain a large amount of peptides and thus has an excellent flavor, richness and richness of flavor, and can be used for general purpose as well as enhancement and persistence of basic taste.

In addition, the present invention can facilitate the maintenance and improvement of health by allowing the easy ingestion by anyone, both young and old, due to the inclusion of fermented soybeans.

In addition, the present invention has an effect of improving the overall efficiency of natural seasoning, thereby maximizing satisfaction and reliability of use for people who cook food or eat cooked food.

Hereinafter, the present invention will be described in detail.

In the method for producing a high-peptide natural seasoning containing fermented soybean of the present invention, soybean is washed more than two times and then washed to remove foreign matter. At this time, the foreign matter-removed soybeans are soaked at 8 ° C for 12 hours.

There is a problem that the taste of the soybean is deteriorated when the temperature and time are out of the above range.

The water-immersed soybeans are supported on the sieve for 1 hour to remove the water. When the water is not removed for 1 hour, the fermentation is not properly performed due to moisture.

The soybeans from which the water has been removed are sterilized in an autoclave (Auto Clave). The soybeans are dried at 110-130 ° C., 1 to 2 atmospheric pressure, and 20 minutes to 40 minutes In the present invention, preferably at 121 DEG C and 1.5 atm for 30 minutes.

If the temperature, the atmospheric pressure, and the time for sterilization are out of the range, sterilization is not properly performed, and fermentation of the raw soybean may not be performed properly.

In the above, the sterilized soybeans are cooled at room temperature or cooled using a refrigerator or the like. The cooled soybeans are placed in a Petri dish on a clean bench, and then B. coagulans ( Bacillus coagulans ) .

In this case, B. coagulans ( Bacillus coagulans ) is inoculated with Petri dish (Petri dish). For example, when 30 g of sterilized soybean is contained in Petri dish, B. coagulans ( Bacillus coagulans ) Is inoculated about 0.9g of 3% of the amount of soybean in petri dish (petri dish).

In this way, fermented soybeans and other natural materials are mixed to produce natural seasonings. The fermented soybeans produced at this time are mixed with radish, cola bean, mushroom, green onion, maize, lotus root, garlic, shrimp, For the optimum combination, fermented soybean was added in an amount of 25.8% by weight, 12.9% by weight, 12.9% by weight of cola, 12.9% by weight of mussel, 6.15% by weight of Maize, 6.15% by weight of Maize, 6.25% 1.26% by weight, clover 6.15% by weight, dodec 6.15% by weight and green pepper 2.13% by weight.

Accordingly, the efficacy and the like can be confirmed through various measurements of the natural seasoning produced by the present invention.

That is, as can be seen from the following Tables, it is possible to measure moisture content, sugar content, pH, pH, reducing sugar, enzyme activity measurement (measurement of protease activity,? -Amylase,? -Amylase) DPPH measurement and free amino acid analysis using LC-MS / MS.

Table 1 below shows the measurement of the moisture content. The powder samples were measured at 105 캜 for 15 minutes using a water activity meter (INFRAREN MOISTURE, Keff, Japan). All samples were measured 3 times repeatedly.

Changes of Moisture contents in white soybean with different  온도 during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 57.1 40.1 36.6 42.5 25.6 30.0 40 ℃ 42.9 25.8 31.0 26.9 22.8 26.2 50 ℃ 27.4 35.7 22.2 10.0 3.8 20.8

5 mg of distilled water was added to 0.5 g of the sample powder, and the sample solution was well dissolved for 30 seconds with shaking more than 50 times. The sample solution was measured three times using a sugar meter (Pockert Refractomer, ATAGO, Japan).

Changes of soluble solids in white soybean with different 온도  during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 1.60 1.87 2.07 2.07 1.27 2.33 40 ℃ 1.13 2.40 1.67 2.47 1.73 1.97 50 ℃ 1.60 1.13 2.20 0.97 1.27 1.13

10 g of fermented soybean and 100 mL of distilled water were pulverized in a grinder and then filtered through 3-fold gauze to measure the pH of soybean fermentation sample. The pH was adjusted to 10 mL of distilled water, 90 mL of distilled water, The samples were homogenized and measured three times using a pH meter.

Changes of pH in white soybean with different 온도 during  fermentation for  72 hour 온도
(%) Fermentation time (hour) 12 24 36 48 60 72 30 ℃ 5.70 5.80 5.85 4.68 4.50 6.11 40 ℃ 5.73 5.58 5.75 5.91 5.96 6.22 50 ℃ 4.25 5.51 5.48 5.92 5.55 5.59

10 g of fermented soybean and 100 mL of distilled water were pulverized in a grinder and then filtered through 3-fold gauze to measure the acidity of soybean fermentation samples. The acidity was measured by adding 0.001 N NaOH Was added and the pH was adjusted to 8.3. The amount of 0.001N NaOH was measured and measured three times.

Changes of asitity in white soybean with different 온도 during  fermentation for  72 hour 온도
(%) Fermentation time (hour) 12 24 36 48 60 72 30 ℃ 24.08 23.29 31.51 36.8 44.16 39.6 40 ℃ 27.54 34.45 24.15 21.945 30.3 34.8 50 ℃ 61.15 23.85 21.02 19.49 21.96 20.92

0.5 ml of the supernatant of soybean was added to the reducing sugar (DNS), and 1.5 ml of the DNS reagent was added thereto. The resulting mixture was incubated in boiling water for 5 minutes, cooled to room temperature, / VIS spectrophotometer to measure the absorbance at 550 nm. At this time, the content was calculated from a standard curve prepared using glucose as a standard sugar, and the results of three repeated measurements were expressed by the average value and the standard deviation. The measured value was expressed in mg / mL.

Changes of Reducing suger in white soybean with different 온도  during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 6.83 14.50 4.83 4.86 3.61 3.65 40 ℃ 2.49 3.80 2.59 1.67 2.93 3.33 50 ℃ 4.64 2.76 3.53 2.57 1.43 1.55

Table 6, Table 7, and Table 8 below show the enzymatic activities. The fermented soybean was pulverized by adding 6 times volume of sodium phosphate buffer (pH 7.2) to 5 g of fermented soybeans, and then pulverized by centrifugation (2000xg, 30 minutes) The supernatant was collected and used to analyze the indicator for the enzyme activity measurement with crude enzyme solution.

That is, Table 5 below shows the measurement of Protease activity during enzyme activity measurement. Protease activity was measured by using the supernatant prepared above as a crude enzyme solution. The substrate solution was prepared by dissolving Hammerstern casein 0.6% in sodium phosphate buffer (pH 7.2). 1 ml of the substrate solution and 1 ml of the crude enzyme solution were placed in a test tube and reacted at 30 ° C for 10 minutes. The reaction was stopped by adding 1 ml of 0.4 M trichloroacetic acid (TCA) solution and left for 30 minutes.

After centrifugation (2000xg, 30 minutes), the supernatant was taken and absorbance was measured at 280 nm using a UV / VIS spectrophotometer. The protease activity was calculated by increasing the enzyme activity of 1 unit by 0.01 / ml / min.

Changes of Protease activity in white soybean with different  온도 during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 0.00 2.70 2.25 2.25 2.14 2.93 40 ℃ -0.63 3.00 2.56 5.08 4.00 4.07 50 ℃ -0.33 1.85 2.27 2.33 2.19 1.75

Amylase activity was measured by dissolving a 0.5% soluble starch solution in 0.4 M acetate buffer (pH 4.8). The results are shown in Table 7 below. 2 ml of the substrate solution and 1 ml of the crude enzyme solution were placed in a test tube and reacted at 30 ° C for 10 minutes. 1 ml of 0.4 M trichloroacetic acid (TCA) solution was added, and the reaction was stopped by allowing to stand for 30 minutes. After removing the precipitate by centrifugation, the supernatant was taken. 10 ml of a solution prepared with 0.005% I ₂ and 0.05% KI was added to 1 ml of the reaction solution, and the absorbance was measured at 600 nm using a UV / VIS spectrophotometer Respectively.

Calculation of the α-amylase activity was calculated by defining the amount of the enzyme whose blank absorbance was decreased by 10% by calculating 1 unit of enzyme activity as an increase of 0.01 / ml / min.

Changes of  α- amylase activity in white soybean with different  온도 during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 2.62 -17.94 -19.30 -19.30 0.52 -18.82 40 ℃ -17.32 -18.90 -14.46 -13.04 -6.12 -14.06 50 ℃ -8.02 -31.50 13.00 -7.00 -3.50 14.82

Amylase activity of β-amylase was measured by dissolving 0.5% soluble starch in 0.4M acetate buffer (pH 4.8). 2 ml of the substrate solution and 1 ml of the crude enzyme solution were placed in a test tube and reacted at 30 ° C for 10 minutes. 1 ml of 0.4 M trichloroacetic acid (TCA) solution was added, and the reaction was stopped by allowing to stand for 30 minutes. The amount of maltose in the supernatant recovered by centrifugation was measured by the DNS method. At this time, the standard curve of the DNS method was created using maltose. Calculation of β-amylase activity was calculated by calculating the amount of enzyme when 1 mg of maltose was liberated in 1 ml of enzyme solution by calculating 1 unit of enzyme activity as an increase of 0.01 / ml / min.

Changes of   β- amylase activity in white soybean with different  온도 during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 0.96 0.34 0.48 0.56 0.58 0.58 40 ℃ 0.47 0.52 0.73 0.57 0.72 0.62 50 ℃ 0.46 0.52 0.78 0.78 1.59 1.63

Amino nitrogen was measured in the following Table 9 and amino nitrogen was measured using a formal titration method. 25 ml of the sample solution was taken with a pipette into the flask, and 25 ml of formal solution and 20 ml of distilled water were added using a measuring cylinder. Blank for blank test was added with 40 ml of distilled water.

About 6 drops of phenolphalein was added thereto and neutralized with 0.1 N NaOH solution until it became pale red.

Amino acid nitrogen (%) = (V ₂ -V ₁ ) × F × 0.0014 × D × S / 100

V ₂ : Proper consumption of test (ml)

V ₁ : Expected consumption of blank test (ml)

F: Potency of 0.1N NaOH

D: Dilution factor

S: Weight of sample (g)

0.0014: the amount of nitrogen (g) equivalent to 1 ml of 0.1 N NaOH solution,

Changes of Amino type nitrogen in white soybean with different  온도 during fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 0.71 0.47 -0.92 -0.61 -0.25 -0.35 40 ℃ -0.61 1.44 0.35 -0.28 -0.18 -0.28 50 ℃ 0.34 -0.48 0.21 0.19 -0.05 0.11

Table 10 below shows the measurement of volatile basic nitrogen (DPPH), and volatile basic nitrogen was measured using the microdiffusion method (Conway method). The microdiffusion method (Conway method) is a thick hard glass similar to a Petri dish and is measured with a diffuser with a modified lid. Tilt the diffuser a little and then add 1 ml of test solution to the bottom of the outer chamber with a pipette and then accurately add 1 ml of 0.01 N sulfuric acid to inner chamber A in the same manner. Apply a small amount of airtight sealant to the replacement part of the cover, quickly put about 1 ml of saturated solution of potassium carbonate in the upper part of the outer chamber B, immediately cover the cover, fix it with a clip, and rotate it while tilting the diffuser back and forth, The test solution and potassium carbonate saturated solution are mixed well and left to stand. Be careful not to mix the solution of the outer chamber with the solution of the inner chamber. And allowed to stand at 25 DEG C for 1 hour.

Open the lid, add a drop of Brinkswick's solution to the inner sulfuric acid solution, and titrate with 0.01 N sodium hydroxide solution using a micro-burette to obtain the average value (a ml) twice. Separately, a blank test was conducted in the same manner using distilled water instead of the test solution, and the average value (b ml) was calculated twice according to the following equation.

mg /% = 0.14 x ((b-a) f / w) 100 d

w: sample weight (g)

a: 0.01 N NaOH consumption (ml)

b: NaOH consumption (ml)

f: 0.01N NaOH

d: dilution factor

Changes of DPPH in white soybean with different 온도 during  fermentation for  72 hour 온도
(%) Fermentation time  ( hour ) 12 24 36 48 60 72 30 ℃ 22.1 22.1 -2.5 75.2 -7.4 0.0 40 ℃ -88.3 -51.5 -14.7 22.1 -29.4 -17.2 50 ℃ -49.0 -68.7 -14.7 -22.1 -19.6 -34.3

Table 12 and Table 13 below show the analysis of free amino acids using LC-MS / MS. Table 12 shows free amino acid analysis of fermented soybean, Table 13 shows free amino acid analysis of fermented soybean, After the measurement of the amount of the sample, ultrasonic extraction was performed at 60 ° C for 30 minutes using distilled water as a solvent. The extract was filtered with a 0.2 μm syringe filter and used as an analytical sample. Quantitative analysis was carried out using amino acid standards as shown in Table 11 below. For amino acid analysis, LC-MS / MS set in positive electrospray ionization ((+) ESI) mode was used. As the analytical column, Inertsil ODS (150 × 4.6 mm, 5 μm) of GL Science (Japan) was used. Nebulizer pressure, N ₂ gas flow rate and temperature were set at 35 psi, 10 L / min, 320 ° C, respectively, and the capillary voltage was maintained at 4 kV. As the mobile phase, Solution A was a 0.1% formic acid solution containing 5 mM ammonium acetate, and Solution B was acetonitrile containing 0.1% formic acid. The solution was started at 0% B at a flow rate of 0.5 mL / min, 100% B, and then decreased to 0% B for 30 minutes. The injection amount of the sample was 5 μl, and the analysis was performed under selected ion monitoring (SIM) conditions as shown in Table 11 below.

The following Table 14 shows sensory evaluation, and the sensory evaluation of the natural seasoning according to the present invention was carried out. 15 g of the natural seasoning according to the present invention was put into 200 ml of soybean soup soup and boiled for 15 minutes in a medium heat. The sample was used as a sensory evaluation sample. 15 g of the seasoning of 'A' known in the market was put into 200 ml of soybean soup, After boiling, it was used as a sensory evaluation sample. The sensory evaluation staff selected 55 students of undergraduate and graduate students in Food and Nutrition Department and explained the purpose and method of experiment in advance. To avoid affecting the sample, the mouth was rinsed before the sample was inspected and crackers were provided with the water. Sensory evaluation items were evaluated for color (appearance), smell, smell, odor, umami, kokumi, sweetness, and overall quality of seasonings.

The evaluation criteria are as follows: 1. Very bad, 2. Bad, 3. Fair, 4. Good 5. Very good.

Exterior The smell of seasoning already Odor Umami
(Delicious taste) Kokumi
(Deep taste) sweetness Overall
quality The natural seasoning of the present invention 4.3 4.4 4.2 4.2 4.3 4.2 4.4 4.3 A company seasoning 4.0 4.2 4.1 4.1 4.1 4.0 4.0 4.1

As shown in Table 14, the natural seasonings produced by the present invention were found to be superior to the commercially available seasonings of the Company A in terms of various evaluation items, and the overall quality was higher than that of the comparative seasonings.

While the present invention has been described in connection with certain exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

Claims (4)

Washing the soaked soybeans with water at a predetermined temperature of 8 ° C for 12 hours, and washing the soaked soybeans with water for 1 hour to remove water, And sterilizing the dehydrated soybeans in an Auto Clave at a temperature of 110 to 130 DEG C, 1 to 2 atm and 20 to 40 minutes, cooling the sterilized soybeans after cooling the method containing the soybean into a Petri dish (Petri dish) in a clean bench, and inoculated with B. coagulans (Bacillus coagulans) bacteria in Petri dishes (Petri dish) of the comprising the steps of producing a fermented soy,
When the fermented soybean was inoculated with B. coagulans ( Bacillus coagulans ) in the Petri dish, 30 g of sterilized soybean in Petri dish was added to B. coagulans ( Bacillus coagulans) ) Was made to inoculate 0.9g of 3% of the amount of soybean in a petri dish (petri dish)
The method for producing fermented soybeans according to any one of claims 1 to 3, wherein the fermented soybeans are produced by mixing fermented soybeans with radish, cola, moss, mackerel, margarine, lotus root, garlic, shrimp, High Peptide Natural Condiments Manufacturing Method.
delete delete The method according to claim 1,
The above fermented soybean contains 25.8% by weight, 12.9% by weight, 12.9% by weight of cola, 12.9% by weight of mussel, 6.15% by weight of broodstock, 6.15% by weight of maize, 6.25% by weight of lotus root, 1.26% by weight of garlic, % By weight, dodec 6.15% by weight, and green pepper 2.13% by weight, based on the total weight of the fermented soybean.