KR101163386B1 - Method for producing Chungkookjang Kochujang with reduced unpleasant odor - Google Patents

Abstract

The present invention is immersed in water and soaked in soybean Bacillus subtilis ( Bacillus subtilis ) and Bacillus rickenformis ( Bacillus) licheniformis ) After inoculating the mixed strain of SCD121067 strain (KCCM 11054P), fermentation at 34 ~ 36 ℃ for 34-38 hours, the fermentation of the fermentation at 4-6 ℃ for 22-26 hours (cheonggukjang) It relates to a production method of the Chungkukjang red pepper paste comprising the step of preparing a powder and the cheongukjang red pepper paste that the unpleasant odor produced by the method is reduced.

Cheonggukjang red pepper paste prepared with the selected fermentation conditions of the present invention showed protease activity, amino nitrogen content and high thrombi solubility and was also excellent sensory.

Bacillus subtilis, Hussook, Cheonggukjang, Aspergillus duck, Cheonggukjang red pepper paste

Description

Method for producing Chungkookjang red pepper paste with reduced unpleasant odor {Method for producing Chungkookjang Kochujang with reduced unpleasant odor}

The present invention relates to a production method of Chunggukjang pepper with reduced odor. More specifically, the mixture of Bacillus subtilis and Bacillus rickenformis SCD121067 strain (KCCM 11054P) by immersing and increasing soybean in water. After inoculating the strain, fermentation at 34-36 ℃ for 34-38 hours, the fermentation of the fermentation at 4-6 ℃ for 22-26 hours (aging) comprising the step of preparing the Cheonggukjang powder, etc. It relates to a production method of kochujang and Cheonggukjang red pepper paste with reduced unpleasant odor produced by the method.

Soybeans, processed soybeans, are known to have high levels of essential nutrients (unsaturated fatty acids, essential amino acids, vitamin K, etc.) and bioactive substances (dietary fiber, isoflavones, trypsin inhibitors, etc.) (Yoon et al., 1984, Korean J. Food Sci.Technol., 16 (4), 357-382). Cheonggukjang (Chang et al., 2005, Korean J. Food Sci.Technol, Vol. 37, No. 2, pp. 255-260), antioxidant effect (Shon et al., 2000, Korean J. Food) Sci.Technol, Vol. 32, No. 4, pp. 936-941), blood pressure lowering and anti-arteriosclerosis (Yang Jeong-Rye et al., 2003, Korean Journal of Food and Nutrition Science 35: 899-905) On the other hand, consumers are avoided because of the unique odor of volatiles (butanoic acid, 3-methyl-butanoic acid) and ammonia produced during fermentation (Lee et al., 2005, Food Science and Industry, Vol. 38, No. . 2). In recent years, many efforts (fermentation strains, fermentation conditions, additives, etc.) have been put in a lot of efforts to reduce the discomfort of Cheonggukjang (Woo et al., 2006, Korean J. Food Preserv .. Vol. 13, No. 1, pp. 77-82).

Kochujang is a fermented food that is unique to Korea, mixed with various ingredients (meju, starch, red pepper powder, etc.) (Hanchang Lee, 2004, Fermented Food Science, Shinkwang Publishing Co.). Recently, research has been conducted to increase the organoleptic function and functionality of kochujang by adding food ingredients (plum, mushroom, garlic oil, etc.) (Lim et al., 2006, Korean J. Food Sci.Technol., Vol. 38, No. 6, pp. 779-784. The taste of kochujang varies according to the manufacturing process (raw materials, blending ratio, manufacturing method and aging conditions) and is affected by microorganisms that grow in Meju or Koji (Cho et al., 1981, Korean J. Food Sci.Technol, Vol. 13, No. 4). Conventional Kochujang produces an unpleasant odor due to the action of microorganisms in Meju (Choi et al., 2000, Korean J. Food Sci.Technol, Vol. 32, No. 1, pp. 125-131). It does not have the deep taste like gochujang.

Therefore, it is necessary to develop a product that maintains the taste as it is, and has less unpleasant odor and maintains the efficacy of Cheonggukjang with the development of a health food that can eat Gochujang and Cheonggukjang at once.

Korean Patent Registration No. 590733 discloses a method for preparing Chungkukjang red pepper paste, which is different from the method of the present invention.

The present invention has been made in accordance with the requirements as described above, the taste of red pepper paste has a quality difference according to the manufacturing process (raw materials, blending ratio, manufacturing method and aging conditions, etc.), and is affected by the microorganisms that grow in Meju or Koji. Therefore, the present invention can improve the quality of Kochujang by adding and mixing the Cheonggukjang during the manufacture of Gochujang, and select the fermentation conditions and fermentation microorganisms of Cheonggukjang in order to reduce unpleasant odor and to cook at low temperature .

In order to solve the above problems, the present invention provides a method for producing Chunggukjang red pepper paste comprising the step of selecting specific fermentation conditions and fermentation microorganisms for the production of Cheonggukjang and ripening at low temperature.

In addition, the present invention provides a cheongukjang red pepper paste that is reduced in odor produced by the above method.

Cheonggukjang red pepper paste made from the Cheonggukjang powder prepared under the selected fermentation conditions of the present invention has reduced discomfort, showed protease activity, amino nitrogen content, high thrombi solubility, and was also excellent sensory.

In order to achieve the object of the present invention,

Soybeans are immersed in water and cooked to increase Bacillus subtilis and Bacillus rickenformis. licheniformis ) inoculated with a mixed strain of SCD121067 strain (KCCM 11054P), followed by fermentation at 34-36 ° C. for 34-38 hours;

Preparing fermented soybean paste powder by aging the fermented product at 4-6 ° C. for 22-26 hours;

Inoculating Aspergillus oryzae on the increased wheat flour, and raising the grain for 46 to 50 hours at 33 to 35 ° C;

Mixing the cooked wheat rice, glutinous rice, brine and the prepared Cheonggukjang powder and aged for 24 to 26 days;

Adding syrup to the saccharified sugars after aging, followed by heat sterilization and adding red pepper powder; And

After cooling, adding spirits and adding sun salt;

It provides a manufacturing method of Cheonggukjang red pepper paste comprising.

Cheonggukjang red pepper paste of the present invention is first produced by soaking soybeans in water and increasing the Bacillus subtilis ( Bacillus subtilis ) and Bacillus rickenformis SCD121067 strain (KCCM 11054P), and then inoculated with a step of fermentation at 34-36 ℃ for 34-38 hours. Immersion is carried out for 20 hours, and steaming is preferably performed at 121 ° C. for 30 minutes. As the fermentation bacteria, it is preferable to use a mixed strain of Bacillus subtilis and Bacillus richenimoform SCD121067 strain (KCCM 11054P), and the mixed strain is preferably inoculated by 1% by weight. Bacillus rickenformis SCD121067 strain was deposited with the Korea Microbiological Conservation Center on November 9, 2009 (Accession No .: KCCM 11054P).

Fermentation conditions using the mixed strain is important, after inoculating the mixed strain, it is fermented for 34 to 38 hours at 34 ~ 36 ℃, preferably at 35 ℃ for 36 hours. Since the taste of gochujang is affected by fermentation conditions and fermentation microorganisms, in the present invention, the unpleasant odor of Cheonggukjang red pepper paste is reduced by specific microorganisms and specific fermentation conditions.

The method of the present invention then comprises the step of ripening the fermented product at 4 ~ 6 ℃ for 22 to 26 hours (dry) and dried to produce the Cheonggukjang powder. One of the important steps to determine the taste of the Cheonggukjang red pepper paste is the ripening process. In the present invention, the ripening process is performed at 4 to 6 ° C. for 22 to 26 hours, preferably at 5 ° C. for 24 hours. Cheonggukjang powder was prepared through the above process, and in the subsequent step, a process of preparing Cheonggukjang red pepper paste using the prepared Cheonggukjang powder will be described.

The method of the present invention then inoculates Aspergillus oryzae into the cooked wheat flour and floats the grain for 46-50 hours at 33-35 ° C. The inoculation amount of the Aspergillus duck material is preferably 0.1% by weight, and the time to float the grain is preferably 48 hours.

The method of the present invention next includes the step of mixing the cooked wheat rice, glutinous rice, brine and the prepared Cheonggukjang powder and aged for 24 to 26 days, preferably the cooked wheat rice, glutinous rice, brine and Mixing the powder to a salt concentration of 7.8% and aging for 25 days. After 25 days of aging, the amino nitrogen content is at least 280 mg%.

Next, the method of the present invention includes the step of adding syrup to the saccharified sugars after aging, and then adding red pepper powder, and the heat sterilization is preferably performed at 65 ° C. for 10 minutes.

The method of the present invention finally comprises the step of adding ethanol after cooling and post-adding sun salt, preferably after cooling to 35 ° C. to add ethanol and adjusting the salt content of the final product to 6.8% It includes the step of post-adding.

Therefore, the production method of the Chungkukjang red pepper paste of the present invention is most preferably

Soybean immersed in water and steamed to inoculate 1% by weight of a mixed strain of Bacillus subtilis ( Bacillus subtilis ) and Bacillus richeniform SCD121067 strain (KCCM 11054P), and then fermented at 35 ℃ for 36 hours;

Aging the fermented product at 5 ° C. for 24 hours and drying to prepare Cheonggukjang powder;

Inoculating 0.1% by weight of Aspergillus oryzae on the increased wheat flour, and then raising the grain at 33-35 ° C. for 48 hours;

Mixing the cooked wheat rice, glutinous rice, brine and the prepared Cheonggukjang powder to a salt concentration of 7.8% and aged for 25 days;

Adding syrup to the saccharified sugars after aging, followed by heat sterilization at 65 ° C. for 10 minutes to add red pepper powder; And

After cooling to 35 ° C., alcohol is added and the salt of the sun salt is added to adjust the salt content of the final product to 6.8%.

The present invention also provides a cheongukjang red pepper paste reduced in the odor produced by the above method. Cheonggukjang red pepper paste made from the Cheonggukjang powder prepared under the selected fermentation conditions of the present invention has reduced discomfort, showed protease activity, amino nitrogen content, high thrombi solubility, and was also excellent sensory.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

1. Experimental material

1) raw material

Soybean used in the production of Cheonggukjang was made by Soon Sunchang Food Co., Ltd. In the preparation of gochujang, wheat flour, starch syrup, red pepper powder, sun salt, wheat rice, spirits, and glutinous rice were used.

2) Used strain

Bacillus subtilis ( Bcillus subtilis) subtilis ) (target food), Bacillus rickenformis ( Bacillus) licheniformis ) SCK121058 (KCCM 11053P) And Bacillus rickenformis ( Bacillus) licheniformis ) SCD121067 (KCCM 11054P) was distributed at the Sunchang Paste Research Center. Bacillus rickenformis SCK121058 strain was deposited with the Korea Microorganism Conservation Center on November 9, 2009 (Accession Number: KCCM 11053P).

Aspergillus duck ash ( Aspergillus oryzae ) (Tokyo Sunchang Food Co., Ltd.) was used as the final product used for the preparation of kochujang.

2. Cheonggukjang Red Pepper Paste Manufacturing Method

1) Cheonggukjang

Cheonggukjang manufacturing process is as shown in FIG. Soybeans were soaked at room temperature (20 hours) and dehydrated (30 minutes) and then steamed (121 ° C, 30 minutes). 100 g of steamed soybeans were added to a Erlenmeyer flask (1000 ml) and sterilized (121 ° C, 15 minutes). Fermented bacteria were incubated in Tryptic Soy Broth (Bacto ^™ , USA) (30 ° C., 15 hours), diluted with sterile distilled water (10 ^{7-10 8} CFU / ml) and inoculated with 1% (w / w) of cooked soybean. The fermentation was carried out for 60 hours (35 ℃, 40 ℃, 45 ℃), and fermentation conditions of the Cheonggukjang after 60 hours after ripening (5 ℃, 10 ℃, 15 ℃). Fermentation strain is a single strain ( Bacillus subtilis , KCCM11053P, KCCM11054P) and mixed strain ( Bacillus subtilis + KCCM11053P, Bacillus subtilis + KCCM11054P, KCCM11053P + KCCM11054P ).

2) Gochujang

Cheonggukjang red pepper paste manufacturing process is as shown in FIG. Aspergillus oryzae was inoculated 0.1% in the increased wheat flour (33-35 ℃, 48 hours). The cooked wheat, glutinous rice, brine (19 Brix) and Cheonggukjang powder were mixed to a salt concentration of 7.8% and aged for 25 days (more than 280 mg% amino acid nitrogen). Starch syrup was added to the completed saccharification, and heat sterilization (65 ° C., 10 minutes) was followed by stirring with red pepper powder. After cooling (35 ° C.), alcoholic beverages were added and sun salt was added to adjust the salt content of the final product to 6.8%.

3. Experimental method

① Fermentation conditions of Cheonggukjang and Boarding  Select condition

1) moisture content

Moisture was analyzed by 105 ° C. drying.

2) pH, pH

The pH was measured by diluting the sample (5 g) with distilled water (45 ml) and shaking at room temperature using a pH meter (ORION Model 420A +, Thermo Orion., USA). The acidity was adjusted to pH 8.3 with 0.1 N NaOH after diluting the sample (5 g) with distilled water (45 ml) and shaking at room temperature. Citric acid content according to the following equation.

Titratable acidity (%) = V x F x A x 100 / W

V: Proper Consumption of 0.1N-NaOH Solution (mL)

F: Titer of 0.1N-NaOH solution

A: amount of organic acid equivalent to 1 mL of 0.1N-NaOH solution (citric acid: 0.0064)

W: sampling amount (g)

3) Chromaticity

For chromaticity, L, a, and b were measured using a color and color difference meter (model SP-80, Japan).

4) amino nitrogen

Amino nitrogen was measured by Formol method. A sample (2 g) is taken in a Erlenmeyer flask (250 ml), distilled water (100 ml) is added, stirred (1 hour), titrated with 0.1N NaOH solution to pH 8.4. Neutral formalin (20 ml) was added thereto and neutralized to pH 8.4 with 0.1 N NaOH solution. A background test was performed on distilled water separately, and amino nitrogen content was obtained according to the following equation.

Amino-type nitrogen =

A: Sample titration amount of 0.1N NaOH solution (ml)

B: blank test (ml) of 0.1N NaOH solution

F: Titer of 0.1N NaOH Solution

5) Ammonia nitrogen

Ammonia nitrogen is Na ₂ of the amino nitrogen and the measurement taken of the same sample solution (0.1 ml) phenol-hypochloride solution A by reaction (10g distilled water, phenol and sodium nitroprusside dihydrate 0.05g of 1,000ml) and B solution (distilled water 1,000ml _{HPO 4? 12H 2 O 9g,} NaOH 6g and NaOCl 10ml) was then put in the reaction, respectively by 2 ml (37 ℃, 20 minutes). The reaction solution was measured for absorbance at 630 nm with a UV spectrophotometer (UV-1650PC, Shimadzu). As the standard curve, (NH ₄ ) ₂ SO ₄ was used.

6) Determination of enzyme activity

(1) Preparation of coenzyme liquid

In the crude enzyme solution, distilled water (190 ml) was added to the sample (10 g), followed by extraction (20 ° C., 3 hours, 150 rpm), followed by centrifugation (Model J2-21 Centrifuge, Beckman LTD., USA) (10000 rpm, 10 The supernatant obtained by distillation) was filtered and the filtrate (Syringe filter, 0.2μ) was used as the crude enzyme solution.

(2) amylase

α-amylase activity was reacted with a 1% soluble starch solution (1 ml) dissolved in coenzyme solution (1 ml) and 0.02M phosphate buffer (pH.6.9) (40 ° C, 30 minutes), followed by 1M acetic acid (10 ml). The reaction was stopped. Iodine solution (0.05% iodine + 0.5% KI) (1 ml) was added to develop color at room temperature and absorbance (660 nm) was measured by UV spectrophotometer. Decrease the absorbance value of blank by 10% as 1 unit and convert it to 1 g of sample.

β-amylase activity was measured by DNS (dinitrosalicylic acid) method. The crude enzyme solution (1 ml) and 0.5% soluble starch solution (1 ml) dissolved in 0.4 M acetic acid buffer (pH 4.8) were reacted (30 ° C., 30 minutes), and then boiled for 5 minutes by adding DNS reagent (1 ml). After boiling, the mixture was cooled quickly and absorbance (540 nm) was measured by adding distilled water (10 ml). Standard calibration curves were prepared using D (+)-maltose and used in multiple dilutions to ensure that the absorbance of the sample was within the effective range. The titer of the enzyme was expressed after 1 ml of the enzyme solution was converted to 1 g of the sample using 1 unit of the amount of enzyme that releases 1 mg of maltose for 30 minutes.

(3) protease

Acidic protease activity was reacted with a crude enzyme solution (1 ml) and a 0.6% casein substrate solution (3 ml) adjusted to pH 3 (30 ° C, 10 minutes), followed by the addition of 0.4M-TCA (Trichloroacetic acid) (5 ml). The reaction was stopped. The reaction solution was allowed to stand (30 ° C, 30 minutes) and filtered (Whatman, No. 2). Take gastric filtrate (2 ml), quickly add 0.4M-Na ₂ CO ₃ (5 ml) and 1N-Folin reagent (Phenol reagent) (1 ml), and develop (30 ° C, 30 minutes) absorbance (660 nm). ) Was measured. For enzyme activity, 1 ml of enzyme solution produced 1 μg of tyrosine in 1 unit for 1 minute, and a calibration curve was prepared using tyrosine as a standard.

Neutral protease activity was adjusted to 0.6% casein substrate solution adjusted to pH 7 in crude enzyme solution (1 ml), and the experimental method was the same as the measurement of acidic protease activity.

7) Sensory test

(1) experimental design and statistical processing

The order of placement and presentation of the samples during sensory evaluation was randomly planned. The mean and standard deviation of each sensory evaluation were calculated using the statistical analysis system (SAS) statistical package (SAS 9.1), and tested for significance by ANOVA analysis (duncan's multiple range test).

(2) Selection of sensory inspectors and sample preparation

The sensory tester consisted of 10 graduate and undergraduate students of food engineering major, Chonbuk National University. For the sensory test of Chungkookjang, raw Cheonggukjang (100 g) was used as a sample. The order of presentation was randomized, and the inspector was provided with the mouth water and the chopsticks needed to evaluate the sample.

(3) Evaluation contents and method

The evaluated items were evaluated with four test items such as color, taste, off-flavor, and preference. In the evaluation, a 9-point scale was used, and the strength was weakened toward 1 point and the strength of the characteristic was increased toward 9 points.

② Quality Analysis of Cheonggukjang Gochujang

1) water content, pH, acidity, amino nitrogen, ammonia nitrogen and enzyme activity

It is the same as the experimental method of selecting the fermentation conditions and ripening conditions of the Cheonggukjang.

2) the number of microorganisms

After diluting the kochujang with physiological saline, it was formed after incubation for 36 hours at 30 ℃ and yeast and mold at 25 ℃ for 72 hours using aerobic bacteria, anaerobic bacteria, yeast, and 3M petrifilm ^™ plates for measuring fungi. Colonies were counted.

3) thrombolytic ability

The strains used for fermentation of Cheonggukjang and the strains of Kochujang Gochujang were cultured in Tryptic Soy Broth (Bacto ^™ , USA) to analyze the thrombolytic ability of the culture. To determine the thrombolytic ability, the red pepper paste was diluted 10-fold, and the thrombus solubility was measured by preparing a bacterium in the dilute pepper paste to be 10 ⁷ CFU / ml. Thrombolytic ability was used a modified method of the fibrin plate method. To 0.3% fibrinogen solution (10 ml) prepared with 50 mM Tris-HCl buffer containing 0.15 M NaCl (pH 7.4) was added 2% agarose (10 ml) prepared in the same buffer. Thrombin (10 units) was added to 0.1% of the whole to form a fibrin clot of uniform thickness and left at room temperature for 1 hour to use for thrombolytic activity. Activity measurement was carried out by placing a paper disc (8mm, Toyo Roshi, Tokyo, Japan) on the fibrin plate prepared, and dropping 20 μl of the strain culture solution to investigate the diameter of the transparent site generated after the reaction (37 ℃, 18 hours), The longest diameter (d ₁ ) and the shortest diameter (d ₂ ) were measured to calculate the area of the lyzed zone (cm ² = p × d ₁ × d ₂ ). The activity of the generated enzyme was expressed as the ratio of plasmin after measuring the area of the lyzed zone formed by adding 20 μl of plasmin (Sigma, St. Louis, MO, USA) to fibrin plates.

4) Sensory test

It is the same as the experimental method of selecting the fermentation conditions and ripening conditions of the Chunggukjang.

Example  1. Selection of fermentation conditions

① Selection of fermentation strain and fermentation temperature

1) moisture content

Changes in water content during fermentation are shown in Table 1. The fermentation rate was 59.04% at the beginning of fermentation. As the fermentation progressed, the water content increased, but decreased later than the fermentation temperature. Mixed strains ( B. subtilis + KCCM11053P and B. subtilis + The 24-hour water content of Cheonggukjang fermented with KCCM11054P) was about 65.20 ~ 62.38%. Mixed strain (KCCM11053P + KCCM11054P) The highest water content was obtained at 35 ° C for 48 hours (60.90%), 36 hours at 40 ° C (63.21%) and 12 hours at 45 ° C (63.23%). The decline happened quickly. Mixed strains ( B. subtilis + KCCM11053P and B. subtilis + KCCM11054P ) showed relatively high moisture content (65.20% and 63.57%) at all temperatures.

Table 1. Changes in water content of Chunggukjang during fermentation time with various fermentation bacteria and temperature.

2) pH and acidity

Changes in pH and acidity during fermentation are shown in Table 2. During fermentation, pH increased from the initial pH (pH 6.56) to 6.73-77.6 (35 ° C), 7.13-77.7 (40 ° C) and 7.43-7.75 (45 ° C) at 60 hours. At 35 ° C, the pH increased as fermentation progressed. At 40 ° C, the pH increased but B. subtilis treatment and mixed strains ( B. subtilis + KCCM11053P and B. subtilis + KCCM11054P) It decreased after 36 hours or 48 hours. In the case of Cheonggukjang fermented at 45 ° C, the pH increased with time, and all strains showed a similar range of pH (7.46 ~ 7.52) at 60 hours. As the fermentation progressed, the pH increased and showed a tendency to decrease after showing the maximum pH.

At 35 ℃, the acidity of Chungkookjang decreased and increased (0.20 ~ 0.40%) after 24 hours (0.11 ~ 0.20%). B. subtilis + KCCM11054P treatment was the lowest at 36 hours of fermentation (0.18%). At 40 ° C and 45 ° C, the acidity of single strains (KCCM11053P and KCCM11054P) and mixed strains (KCCM11053P + KCCM11054P ) increased rapidly up to 12 hours (0.47 ~ 0.56%) after fermentation started but then gradually decreased.

Table 2. Changes in pH and acidity of Cheonggukjang during fermentation time with various fermentors and temperature.

3) Chromaticity

Changes in chromaticity during fermentation are shown in Tables 3 to 5. L and b of Chonggukjang decreased as fermentation progressed and a increased. KCCM11053P at 35 ℃, 60 hours Treatments (30.94, 13.21) showed lower L and b than B. subtilis (39.43, 14.61) and KCCM11054P (35.66, 13.94). KCCM11053P The mixed strains ( B. subtilis + KCCM11053P and KCCM11053P + KCCM11054P ) also showed low L (34.34 ~ 29.02) and b (13.85 ~ 12.01). As a result of measuring the fermentation temperature differently, the change of L, a, b could not be observed largely according to the treatment.

Table 3. Changes in chromaticity of Cheonggukjang during fermentation time according to various fermentors at 35 ° C.

Table 4. Changes in chromaticity of Cheonggukjang during fermentation time at various fermentors at 40 ° C.

Table 5. Change in chromaticity of Cheonggukjang during fermentation time according to various fermentation bacteria at 45 ℃.

5) amino nitrogen

Amino nitrogen change during the fermentation of Cheonggukjang is shown in FIG. The single strains (KCCM11053P and KCCM11054P ) were higher than the B. subtilis treatments, and Cheonggukjang prepared with mixed strains (KCCM11053P + KCCM11054P) showed the highest amino nitrogen content. KCCM11053P for 60 hours of fermentation at 35 ℃ Treatment (586.25 mg%) was KCCM11054P Amino nitrogen content was lower than the treatment (597.1 mg%) and B. subtilis + KCCM11053P treatment (351.40 mg%) was also treated with B. subtilis + KCCM11054P. Treatment (486.50 mg%) Lower content was shown. Similar trends were observed at 40 ° C. KCCM11053P at 60 hours of fermentation Treatment (696.85 mg%), KCCM11054P Treatments (817.6 mg%), B. subtilis + KCCM11053P treatment (462 mg%) and B. subtilis + KCCM11054P treatment (493.15 mg%) are shown. The amino nitrogen content continued to increase until 72 hours after the fermentation of Cheonggukjang and the production of amino nitrogen was different according to the fermentation strains, Lee et al. (1991, Korean J. Food Sci. Technol, Vol. 23, No. 4, pp. 478-484). In case of B. subtilis + KCCM11054P treatment, the content increased as the temperature was 486.50 mg% (35 ℃), 493.15 mg% (40 ℃) and 527.10 mg% (45 ℃) at 60 hours.

6) Ammonia nitrogen

Ammonia nitrogen change during fermentation of Cheonggukjang is shown in FIG. 4. Ammonia nitrogen content increased with the fermentation time, which is Bacillus et al., Seok (1994, Korean Soc. Appl. Biol. Chem., Vol. 37, No. 2, pp. 65-71) Cheonggukjang using licheniformis CN-115 strain was almost constant at the beginning of fermentation, which increased after 24 hours and reached the highest content at 60 hours. The fermentation started and gradually increased until 12 hours (4.75 ~ 11.98 mg%). After 12 hours, the content of ammonia nitrogen increased rapidly. After 48 hours, all strains except for B. subtilis treatment (45.82 ~ 46.38 mg%) showed similar ammonia nitrogen content (70.88 ~ 82.65 mg%). At 60 hours of fermentation, B. subtilis treatment (46.38 mg%) was the lowest and KCCM11054P treatment (88.45 mg%) was the highest. Cheonggukjang fermented for 60 hours showed ammonia nitrogen content in the range of 82.40 ~ 87.71 mg% (40 ℃) and 84.23 ~ 87.48 mg% (45 ℃).

7) enzyme activity

(1) amylase activity

① α-amylase activity

Changes in α-amylase activity during fermentation were as shown in FIG. 5. It showed a tendency to increase as the fermentation proceeds at 35 ℃. B. subtilis at 60 hours of fermentation Treatments showed 6.70 unit / g and single strains (KCCM11053P and KCCM11054P ) and mixed strains ( B. subtilis + KCCM11053P, B. subtilis + KCCM11054P and KCCM11053P + KCCM11054P) ranged from 8.93 to 9.94 units / g. Α-amylase activity increased with fermentation time at 40 ° C and then decreased after 48 hours (8.40-9.24 unit / g). B. subtilis at 45 ℃ Treated and mixed strains ( B. subtilis + KCCM11053P and B. subtilis + KCCM11054P) increased (7.61 ~ 7.91 unit / g) as fermentation progressed, but single strains (KCCM11053P and KCCM11054P ) and mixed strains (KCCM11053P + KCCM11054P ) It decreased after -48 hours (0-2.82 unit / g).

② β-amylase activity

Β-amylase activity change during the fermentation of Cheonggukjang is shown in FIG. The initial fermentation of 0.22 unit / g at 35 ℃ showed a rapid increase up to 12 hours (36.52 ~ 60.22 unit / g) and then decreased as the fermentation progressed. Similar trends were observed at 40 ° C and 45 ° C. Oh (2008, Korean J. Food Sci. Technol, Vol. 40, No. 1, pp. 56-62) showed a rapidly increased β-amylase activity up to 12 hours in germinated soybean Chungkukjang, similar to the report decreased after 24 hours. It was. β-amylase activity was highest in KCCM11053P + KCCM11054P treatment at 35 ° C (60.22 unit / g) at 12 hours of fermentation and B. subtilis Treatments were lowest at 45 ° C. (16.52 unit / g). B. subtilis + KCCM11054P treatment showed high activity at 35 ° C with 52.81 unit / g (35 ° C), 48.37 unit / g (40 ° C) and 36.52 unit / g (45 ° C) at 12 hours of fermentation.

(2) protease activity

① acid protease

The acidic protease activity change during fermentation of Chunggukjang is shown in FIG. At 35 ° C., acidic protease activity increased with fermentation time. It increased more than 2 times from 13.92 unit / g at the beginning of fermentation to 32.79 unit / g ( B. subtilis treatment) at 48 hours of fermentation. Acidic protease activity by strain showed high acidic protease activity in all strains (48.59 ~ 77.67 unit / g) except B. subtilis treatment (33.15 unit / g). Cheonggukjang fermented at 40 ℃ showed similar changes in acidic protease activity at 35 ℃. Similar trend was also observed at 45 ℃, but increased slowly up to 24 hours (17.26 ~ 27.92 unit / g) and then increased rapidly after 36 hours (24.23 ~ 44.85 unit / g). Acidic protease activity of 60 hours of fermentation showed that B. subtilis treatment showed the highest activity at 40 ° C (37.26 unit / g), and single strain (KCCM11053P and KCCM11054P ) and mixed strain ( B. subtilis + KCCM11053P, B. subtilis + KCCM11054P and KCCM11053P + KCCM11054P) showed the highest activity at 35 ° C (48.59 ~ 77.67 unit / g).

② neutral protease

Neutral protease activity changes during fermentation in Chunggukjang are shown in FIG. 8. As fermentation progressed, neutral protease activity increased. B. subtilis at 35 ° C. (36 h) The treatment showed the lowest activity at 12.59 unit / g and the KCCM11053P treatment showed the highest activity at 56.95 unit / g. B. subtilis All strains (48.59 ~ 77.67 unit / g) except the treatment (33.15 unit / g) showed high activity at 35 ℃. When B. subtilis + KCCM11054P was fermented for 36 hours, it showed 20.00 unit / g (35 ℃), 18.28 unit / g (40 ℃), 18.24 unit / g (45 ℃), showing the highest neutral protease activity at 35 ℃. appear.

8) Sensory test

The results of sensory evaluation to select the fermentation conditions of Cheonggukjang are shown in Table 6. In order to investigate the sensory function of each strain, each bacteria were inoculated and fermented for 36 hours and then sensory test was performed with Saengcheonggukjang. B. subtilis is used as a control Single strain ( B. subtilis , KCCM11053P KCCM11054P ) And mixed strains ( B. subtilis + KCCM11053P, B. subtilis + KCCM11054P, and KCCM11053P + KCCM11054P ) compared with the Cheonggukjang made the color was the highest preference of KCCM11053P treatment. Taste is B. subtilis + KCCM11054P treatment received the highest rating and KCCM11053P treatment received the lowest rating. In the off-flavor, B. subtilis + KCCM11054P treatment was also evaluated to have the lowest off-flavor with the lowest rating. B. subtilis + KCCM11054P> B. subtilis + KCCM11053P> B. subtilis > KCCM11054P > KCCM11053P + KCCM11054P> KCCM11053P . Overall, B. subtilis + KCCM11054P treatment scored highest for most items, including aroma, taste, and overall preference. Therefore, it was evaluated that B. subtilis + KCCM11054P treatment was effective in terms of sensory improvement as a result of fermentation strain selection.

The results of sensory evaluation according to fermentation temperature of Cheonggukjang are shown in Table 7. The color of the sample at 45 ° C. received the highest rating and the taste of the sample fermented at 35 ° C. received the highest rating. The off-flavor was evaluated to have the lowest off-flavor, with Cheonggukjang fermented at 35 ℃. The overall preference was also highest in Cheonggukjang fermented at 35 ℃.

Therefore, the results of sensory evaluation in screening strains and fermentation temperature in order to reduce the discomfortB. subtilis + It is effective to use KCCM11054P (35 ℃, 36 hours) as a sample of Cheonggukjang red pepper paste.

Table 6. Sensory Characteristics of Cheonggukjang by Fermentation Bacteria.

Table 7. Sensory Characteristics of Cheonggukjang with Different Fermentation Temperatures.

② Cheonggukjang Boarding  Select condition

Fermented strain ( B. subtilis + KCCM11054P), fermentation time (36 hr) and temperature (35 ℃) were selected to reduce the odor of Cheonggukjang. Fermented Cheonggukjang were ripened for 5 hours at 5 ° C, 10 ° C and 15 ° C for 60 hours and analyzed at 12 hour intervals.

1) moisture content, pH and acidity

The water content decreased gradually with fermentation time and increased slightly after 36 hours, but no noticeable change was observed.

The pH increased as the ripening progressed at 5 ° C., and peaked at 24 hours (pH 7.52), and then decreased. The ripened pH also increased at 10 ° C and 15 ° C and then decreased after peaking at 48 h (pH 7.69-7.72). As the ripening time, the pH increased and then decreased.

Acidity decreased from 0.18% at early stage to 12 hours (0.43 ~ 0.48%). As the ripening progressed at 15 ° C., it decreased to 48 hours (0.29%) and then increased again.

Table 8. Changes in pH of Cheonggukjang during fermentation time at various temperatures.

2) amino nitrogen and ammonia nitrogen

The amino nitrogen content was found to increase more than twice as early as ripening. At 12 ° C., 15 ° C. (536.20 mg%) ripening showed the highest amino nitrogen content. At 5 ° C and 10 ° C, the amino nitrogen content increased with ripening and then decreased after 48 hours (616.70 mg%, 642.25 mg%). At 15 ° C. increased during ripening but decreased slightly at 48 hours (540.75 mg%) and then increased again (614.95 mg%).

Ammonia nitrogen increased rapidly at 12 hours (85.59 to 84.12 mg%) but slightly decreased at 24 hours (79.82 to 83.92 mg%) and then remained constant. The higher the ripening temperature, the higher the ammonia nitrogen content, and the lowest value of ammonia nitrogen content was found in the Cheonggukjang after 24 hours (79.82 mg%) at 5 ° C.

3) enzyme activity

(1) α-amylase activity and β-amylase activity

α-amylase activity was slightly increased as Chonggukjang, which was 6.90 units / g at 0 hours, but showed constant activity after 24 hours (8.97 ~ 9.52 unit / g). β-amylase activity decreased as the ripening progressed. 26.89 unit / g at 0 hours and then β-amylase activity in the range of 2.44 to 0.22 unit / g at all ripening temperatures.

(2) acidic protease activity and neutral protease activity

The acidic protease activity of Cheonggukjang ranged from 39.15 to 43.46 unit / g during ripening, with no significant change. Neutral protease activity increased after 0 hours (20.08 unit / g). After 24 hours (33.32 ~ 35.08 unit / g) slightly increased, the higher the ripening temperature, the higher the neutral protease activity.

4) Sensory evaluation

Cheonggukjang's sensory evaluation after boarding is shown in Table 9. After fermenting the Cheonggukjang, the sensory evaluation was performed using the Cheonggukjang after ripening by temperature and the least unpleasant smell at 5 ℃. In order to select the ripening period, sensory evaluation was performed using samples ripened for 0 to 36 hours. The taste was generally reduced but when bitterness was over 36 hours, the bitter taste was strongly felt. As a result, the color showed high rating in the order of 24 hours> 36 hours> 12 hours> 0 hours. Taste was high in order of 24 hours> 12 hours> 0 hours> 36 hours. The discomfort according to the boarding time was evaluated to be the lowest for the 24-hour boarding Cheonggukjang. Overall preferences were better than those of the lesser Chungkookjang. 24 hours> 12 hours> 36 hours according to the ripening time.

Table 9. Sensory Characteristics of Cheonggukjang with Fermentation Time at 5 ° C.

Fermentation ( B. subtilis + KCCM11054P , 35 ℃, 36 hours) and ripening (5 ℃, 24 hours) conditions were selected to reduce the odor of Cheonggukjang red pepper paste. This Cheonggukjang was dried and powdered to prepare Cheonggukjang Gochujang.

② Quality Analysis of Cheonggukjang Gochujang

1) Analysis of Physicochemical Quality

Comparison of the quality characteristics of Cheonggukjang Kochujang prepared by Toksun Soon Food Co., Ltd. and Cheonggukjang Kochujang prepared by Chonbuk National University of the present invention are shown in Table 10. Cheongukjang red pepper paste prepared by Chonbuk National University of the present invention was high in water content (47.27%), pH (pH 4.81) and acidity (1.28%). It also had a high amino nitrogen content (420 mg%) and good neutral and acidic protease activity (73.24 unit / g, 92.29 unit / g). Nitrogen content of ammonia nitrogen was higher in Kochujang (31.98 mg%), Cheonggukjang, Chonbuk National University.

Table 10. Physicochemical Properties of Cheonggukjang Red Pepper Paste.

2) thrombolytic ability

The results of investigating the thrombolytic activity using the microorganisms used in Cheonggukjang red pepper paste are shown in Table 11 and FIG. 12. Microorganisms were inoculated in Tryptic Soy Broth (Bacto ^™ , USA) medium and cultured, and then the thrombolytic activity was measured using the fibrin plate method using the culture medium. The strain of Cheonggukjang red pepper paste prepared by Chonbuk National University (266.80%) showed higher thrombolytic activity than the Cheonggukjang red pepper paste strain (156.64%) manufactured by Tosun Sunchang Food Co., Ltd. The thrombolytic activity of Cheonggukjang pepper was higher than that of Cheonggukjang strains (147.66%, 177.34%).

Table 11. Thrombolytic Activity of Fermented Bacteria and Chungkukjang Pepper.

3) Sensory test

Table 12 shows the results of sensory evaluation of Cheonggukjang Gochujang prepared by Indigo Sunchang Foods Co., Ltd. and Cheonggukjang Gochujang prepared by Chonbuk National University. In terms of color, taste, and overall preference, Cheongbukjang red pepper paste was judged to be good, and off-flavor was evaluated as low.

Table 12. Sensory Characteristics of Cheonggukjang Red Pepper Paste.

As a result of the analysis of Cheonggukjang red pepper paste, Cheonggukjang red pepper paste made from Cheonggukjang prepared by Chonbuk National University of the present invention has high protease activity and amino amino nitrogen content and showed high thrombolytic ability. It was also excellent sensory.

1 is a process of manufacturing Chungkookjang.

2 is a manufacturing process of the Cheonggukjang red pepper paste.

Figure 3 shows the change in amino nitrogen content of Chunggukjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

Figure 4 shows the change in ammonia-like nitrogen content of Chunggukjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

5 shows changes in α-amylase activity of Chungkookjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

Figure 6 shows the change of β-amylase activity of Chunggukjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

Figure 7 shows the change in acidic protease activity of Cheonggukjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

Figure 8 shows the change in the neutral protease activity of Cheonggukjang during fermentation time according to various fermentation bacteria and temperature. A: 35 ° C, B: 40 ° C, C: 45 ° C

Figure 9 shows the change of amino nitrogen and ammonia nitrogen of Cheonggukjang during fermentation time according to various temperatures. A: amino nitrogen, B: ammonia nitrogen

10 shows changes in α-amylase activity and β-amylase activity of Chungkookjang during fermentation time according to various temperatures. A: α-amylase activity, B: β-amylase activity

11 shows changes in acidic protease activity and neutral protease activity of Chunggukjang during fermentation time according to various temperatures. A: acidic protease activity, B: neutral protease activity

12 shows the thrombolytic activity of fermented bacteria and Chungkukjang red pepper paste as assessed by the size of the lysed zone on the fibrin plate. A: control, B: B. subtilis , C: KCCM11054P , D: Tobagi, E: CNU

Claims (3)

Soybeans were soaked in water and steamed to inoculate 1% by weight of a mixed strain of Bacillus subtilis and Bacillus licheniformis SCD121067 strain (KCCM 11054P), followed by fermentation at 35 ° C. for 36 hours. Doing; Aging the fermented product at 5 ° C. for 24 hours and drying to prepare Cheonggukjang powder; Inoculating 0.1% by weight of Aspergillus oryza (Aspergillus oryzae) in the increased wheat flour, to produce a Aspergillus ducky Koji (Aspergillus oryzae Koji) by floating the grain for 48 hours at 33 ~ 35 ℃; Mixing the prepared Aspergillus orijae Koji and the increased wheat, glutinous rice, saline and the prepared Cheonggukjang powder to a salt concentration of 7.8% and aged for 25 days; Adding syrup to the saccharified sugars after aging, followed by heat sterilization at 65 ° C. for 10 minutes to add red pepper powder; And After cooling to 35 ° C., adding alcohol and then adding sun salt to adjust to a salt content of 6.8% of the final product; Cheonggukjang pepper production method comprising a. delete Cheonggukjang red pepper paste reduced odor produced by the method of claim 1.

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