KR20160073020A - Manufacturing method of peptide natural seasoning containing microorganism white soybean - Google Patents

Abstract

The present invention relates to a method for preparing a high peptide natural seasoning containing fermented soybean (white bean) comprising: a step of straining soybean (white bean) two or more times and washing the soybean (white bean) to remove foreign substances; a step of water-soaking the soybean (white bean) from which the foreign substances are removed at a constant temperature and for a certain period of time; a step of removing moisture by putting the water-soaked soybean (white bean) in a colander for 1 hour; a step of inserting the soybean (white bean) from which moisture is removed, in an autoclave to sterilize the soybean (white bean) at a constant temperature, at a constant pressure, and for a certain period of time; a step of cooling the sterilized soybean (white bean) and inserting the cooled soybean (white bean) in a Petri dish on a clean bench; and a step of preparing fermented soybean (white bean) by inoculating B. coagulans (Bacillus coagulans) bacteria to the Petri dish. In addition, according to the present invention, in the step of preparing fermented soybean (white bean), the prepared fermented soybean (white bean) is mixed with white radish, kohlrabi, button mushroom, spring onion, yam, lotus root, garlic, shrimp, baby clam, Codonopsis lanceolata, 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 method for producing a high-peptide natural seasoning containing fermented soybean (white rice)

More specifically, the present invention can be applied to various foods by producing natural seasoning containing fermented soybean (white rice), thereby enhancing the richness and richness of the richness and richness of the richness and the richness of the natural seasoning, And a fermented soybean (white rice paddy) so as to obtain a high-fat 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 shellfish 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 Publication No. 10-2006-0003858, which has been conventionally conducted, discloses a method for dissolving wheat gluten enzyme-decomposing seasoning in water to provide a sugar peptide and a peptide having a rich taste-imparting action, separating the resulting aqueous solution by an ultrafiltration membrane 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 which has a high content of peptides by using a natural raw material containing fermented soybean (white rice paddy), which is excellent in a richness of taste, rich in taste and rich in sensation, And a fermented soybean (white rice).

It is another object of the present invention to provide a high-peptide natural food containing fermented soybeans which maximizes the overall efficiency of the natural seasoning and maximizes the satisfaction and reliability of utilization for people who cook food or eat cooked food And a method for producing seasonings.

In order to solve the above-mentioned problem, the present invention provides a method for producing a high-peptide natural seasoning containing fermented soybean (white rice) according to the present invention comprises the steps of washing soybean (white rice) A step of soaking the removed soybean (white rice) at a predetermined temperature and for a predetermined period of time; supporting the soaked soybean (white rice) on the sieve for 1 hour to remove moisture; Is placed in an autoclave and sterilized at a predetermined temperature, a constant atmospheric pressure and a predetermined time, and a step of cooling the sterilized soybean (white rice) and then cooling the soybean (white rice) into a glass container in a clean bench And a step of inoculating B. coagulans ( Bacillus coagulans ) into the glass container to prepare fermented soybean (white rice).

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 method for producing fermented soybean (white rice), which comprises mixing the fermented soybean (white rice) with the radish, cola bean, mushroom, green onion, maize, lotus root, garlic, shrimp, , 12.9% by weight of cola, 12.9% by weight of mussel, 12.9% by weight of mussel, 6.15% by weight of Maize, 6.15% by weight of Maize, 6.25% by weight of Lotus root, 1.26% by weight of garlic, 1.26% by weight of shrimp, 6.15% by weight of clams, 6.15% by weight of dodeca and 2.13% by weight of green pepper.

The present invention uses a natural raw material containing fermented soybean (white rice) to provide a large amount of peptides and thus has excellent fatness, richness of flavor and richness, enhanced basic taste and sustainability, .

In addition, since the present invention includes fermented soybean (white rice), it can be easily ingested by anyone of all ages, thereby helping maintain and improve health.

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 (white rice) according to the present invention, it is washed twice after the white rice (soybean) is washed more than twice, and then the foreign substance is removed. At this time, the soybean (white rice) from which the foreign substance is removed is soaked at 8 ° C for 12 hours.

If the temperature and time are out of the above range, there is a problem that the taste of white rice (soybean) is deteriorated.

The water-immersed white rice (soybean) is supported on the sieve for 1 hour to remove the water. In this case, if 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 which is a sterilizer. The soybeans are dried at 110-130 ° C., 1-2 atmospheric pressure, 40 minutes, and in the present invention, it is preferably carried out 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 (white rice) may not be properly performed.

In the above, the sterilized soybean (white rice) is cooled at room temperature or cooled using a refrigerator or the like, and the cooled white rice is placed in a petri dish in a clean bench and then inoculated with B. coagulans ( Bacillus coagulans ) Fermented soybeans are produced.

In this case, B. coagulans ( Bacillus coagulans ) is inoculated with Petri dish (Petri dish) in a certain amount. For example, when 30 g of the white mold is sterilized in Petri dish, B. coagulans ( Bacillus coagulans ) Is inoculated with 0.9% of 3% of the amount of white spot in the petri dish (Petri dish).

In this way, fermented soybean (white rice) and other natural materials are mixed to produce a natural seasoning. The fermented soybean (white rice) produced at this time is mixed with radish, cola, mushroom, green onion, , 12.8% by weight, 12.9% by weight of cola, 12.9% by weight of mussel, 6.15% by weight of mussel, 6.15% by weight of mussel, 6.15% by weight of lotus root, It is preferable to mix them in a ratio of 6.25% by weight, 1.26% by weight of garlic, 1.26% by weight of shrimp, 6.15% by weight of clam, 6.15% by weight of doduck and 2.13% by weight of green pepper.

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

Table 3 below shows the pH values of the fermented soybeans and 100 mL of distilled water. The pH of the fermented soybeans was measured using a three-ply gauze filter. A sample made by adding 90 mL of distilled water to 10 mL of the solution was 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 (white rice) was used as the reducing sugar, and 1.5 ml of the DNS reagent was added thereto. The resulting mixture was reacted with boiling water for 5 minutes, After cooling, the absorbance was measured at 550 nm using a UV / VIS spectrophotometer. 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 activity measurements. The fermented soybean (5 g) was pulverized by adding 6 times volume of sodium phosphate buffer (pH 7.2) and centrifuged ), And the supernatant was recovered and used for the analysis of the indicator for 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 decreased by 10% by calculating 1 unit of the 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 to the solution, and the solution was 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 ((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 by LC-MS / MS. Table 12 is the analysis of free amino acids of unfermented soybean (white rice), and Table 13 shows the results of analysis of free amino acids of fermented soybean Amino acid was analyzed. Ultrasonic extraction was performed for 30 minutes at 60 ° C using distilled water as a solvent after measuring a predetermined amount of the sample. 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. Normal, 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 soybeans with the soybeans at least two times and washing them to remove foreign matter;
Immersing the soybean (white rice) from which foreign substances have been removed for a predetermined time and for a predetermined time;
Supporting the water-immersed soybeans (white rice bran) on the sieve for 1 hour to remove moisture;
The method of the present invention comprises the steps of putting soybeans from which moisture has been removed in an auto clave and sterilizing the soybeans at a constant temperature and a constant atmospheric pressure for a predetermined period of time; ) In a Petri dish on a clean bench; And
A method for preparing a high-fat natural seasoning containing fermented soybeans, comprising the step of inoculating B. coagulans ( Bacillus coagulans ) into Petri dish to produce fermented soybean (white rice).
The method according to claim 1,
In the steeping step, the temperature is 8 DEG C and the time is 12 hours,
Wherein the sterilization is performed at a temperature of 110 to 130 ° C, an atmospheric pressure of 1 to 2 atmospheres, and a time of 20 minutes to 40 minutes.
The method according to claim 1,
The fermented soybean (white rice) is produced by mixing the fermented soybean (white rice) with the radish, cola bean, mushroom, green onion, margarine, lotus root, garlic, shrimp, clam, ≪ / RTI > wherein the fermentation zone comprises a fermentation zone.
The method of claim 3,
The above fermented soybean (white rice) 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 broiler, 6.15% by weight of maize, 6.25% by weight of lotus root, 1.26% , 6.15% by weight of clams, 6.15% by weight of dodeca and 2.13% by weight of green pepper.