KR100903186B1 - A preparation method of chungkukjang using Bacilluse licheniformis - Google Patents

Abstract

The present invention relates to a novel method for producing a flavored Cheonggukjang using a Bacillus strain, according to the present invention is the molecular weight of the main protein is 19,000 (MW) and the water-soluble amino acids such as cystine and methionine related to the odor of Cheonggukjang is lost There is an excellent effect of providing a new Cheonggukjang with improved flavor.

Cheonggukjang, fermented food, soybean, Bacillus

Description

A preparation method of chungkukjang using Bacilluse licheniformis}

The present invention relates to a new method of manufacturing a flavor of the improved taste using Bacillus strains, and more specifically, the addition of Bacillus licheniformis strain during the production of Cheonggukjang cystine and methionine, etc. The present invention relates to a method for producing a new Chungkookjang with improved flavor by the loss of water-soluble amino acids.

Cheonggukjang is known as one of the unique fermented foods in Korea, which is a major seasoning, corrosion and even amino acid.

Park and Kim et al. Identified and identified effective bacteria during fermentation of Cheonggukjang, and many researchers examined the optimal conditions for the production of Cheonggukjang. There have been reports of changes in amino acids, peptides, small intestine, volatile fragrance components, sugars and oils and fats produced during the production of Cheonggukjang Meju. Chung et al. Investigated the role of strains having phytase activity in the production of Chunggukjang. Cheonggukjang was prepared using natto ) to investigate the tissue and chemical properties of the product.

The like are reviewed Bacillus NATO (B. natto) and Bacillus subtilis were prepared in two soybean strains (B. subtilis) viscosity, color and spreadability, etc. to prepare a processed food in the form of a spread in the soybean material The physicochemical characteristics of viscous substances and bioactive groups produced during fermentation process were investigated.

As mentioned above, B. natto and B. subtilis in the production of most of Cheonggukjang There have been reports of studies on Cheonggukjang using two strains, but no studies on the production of Cheonggukjang using B. licheniformis strains have been reported.

An object of the present invention was devised in view of the above point is to provide a Cheonggukjang that is water-soluble amino acids related to the smell of Cheonggukjang using the Bacillus Richenomymis strain.

The above object of the present invention was achieved by confirming that the contents of water-soluble amino acids cystine and methionine are lost by adding Bacillus licheniformis strain during the preparation of Cheonggukjang.

Hereinafter, the configuration of the present invention will be described.

The present invention provides a novel method for producing a cheongukjang with improved flavor of water-soluble amino acids associated with the inherent odor of cheonggukjang.

The present invention is characterized in that Bacillus licheniformis ( Bacillus licheniformis ) strain is added as a fermentation strain during the production of Cheonggukjang.

Cheonggukjang of the present invention can be prepared by inoculating Bacillus licheniformis strain in the soybeans increased and fermented at 35 ~ 38 ℃ for 36-60 hours, the prepared Cheonggukjang is a water-soluble amino acid associated with the unique smell of Cheonggukjang lost The flavor is improved.

Bacillus licheniformis (Bcillus licheniformis ) CY-1480 was used as a strain for the production of Cheonggukjang in the present invention, isolated and identified in Yeungnam University Biochemistry Laboratory, and deposited with KACC 91325P on August 22, 2007. Strain.

Soybean used in the present invention was used in Korea Taebaek soybean oil and the general components of the soybean oil was crude protein 35.65%, crude fat 19.10%, crude ash 5.35%, moisture 8.99%.

The present invention has a very excellent effect of providing a method for producing a new Cheonggukjang improved flavor by the addition of Bacillus licheniformis strain in the production of Cheonggukjang to lose the content of the water-soluble amino acid cystine related to the cheonggukjang intrinsic odor have.

Hereinafter, an Example demonstrates this invention in detail. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example  1. Cheonggukjang Manufacturing

Cheonggukjang was prepared by Kim et al. 3 kg of selected soybeans were soaked in water at 20 ℃ for 15 hours, drained for 1 hour, weighed about 200 g in 300 mL flask, and then sprinkled for 1 hour at a pressure of 10 lbs in autoclave. It was. The auto-craft soybean was cooled to 50 ° C. and inoculated with Bacillus licheniformis CY-1480 (KACC 91325P) strain under aseptic conditions to incubate for 36-60 hours at 35-38 ° C. to prepare Chungkookjang. It was.

Experimental Example  1. Measurement of general ingredients

Moisture, crude protein, crude fat, crude fiber, pH, titratable acidity and ash content of Cheonggukjang added with the present invention Bacillus Richenomyces strain prepared in the above example was measured by AOAC method, and amino nitrogen was determined by using a foam titration method and ammonia. Nitrogen was measured by the folin method.

As a result of analyzing the moisture, crude protein, crude fat, crude fiber, crude ash, amino nitrogen, ammonia nitrogen in the fermentation process of Cheonggukjang added Bacillus Richenomyces strain of the present invention.

The water content tended to decrease with the fermentation time at 52.43% immediately after the increase, but no significant difference was observed. As the fermentation progressed, the contents of crude protein and crude fat showed a somewhat irregular increase and decrease, and the contents of crude protein and crude fat were almost unchanged compared to raw soybeans. Amino-type nitrogen is an important factor of the taste factor of Cheonggukjang, which is unchanged from the beginning of fermentation to 12 hours, and then rapidly increases to 14.6 mg / g at 48 hours of fermentation, and 18.1 mg / g at 60 hours of final fermentation. Increased.

Analysis of General Components in Fermentation Process of Cheonggukjang division Fermentation period (hours) 0 6 12 24 36 48 60 moisture(%) 52.5 53.3 49.4 49.7 48.7 46.8 46.1 % Viewed 2.3 2.1 2.2 2.2 2.3 2.3 2.1 Crude fat (%) 7.8 7.4 8.3 8.6 8.1 7.9 7.8 Crude Protein (%) 17.8 17.5 17.4 18.8 17.7 17.4 17.3 Crude fiber (%) 4.9 6.6 5.4 5.8 5.8 4.7 4.8 Amino nitrogen (%) 0.28 0.30 0.35 0.35 0.612 14.59 18.1 Ammonia Nitrogen (%) 0.42 0.70 0.70 0.70 0.91 0.99 1.27

Experimental Example  2. Changes in pH and titratable acidity

The changes of pH and titratable acidity during the fermentation process of Cheonggukjang added Bacillus licheniformis strain of the present invention are shown in Table 2. Immediately after the increase of pH, the pH was 6.23. As the fermentation progressed, the pH rose to 8.39 at 60 hours. The titratable acidity gradually decreased after 24 hours with the fermentation time.

Changes of pH and Proper Acidity during Cheonggukjang Fermentation division Fermentation period (hours) 0 6 12 24 36 48 60 pH 6.25 6.30 6.52 6.84 7.49 8.10 8.35 Titratable acidity 0.45 0.50 0.40 0.65 0.50 0.30 0.26

Experimental Example  3. Changes in Protease Activity

In the present invention, 10 g of Cheonggukjang added Bacillus licheniformis strain was added, distilled water was added to 100 mL, shaken and extracted at room temperature for 1 hour, and the filtrate was used as a coenzyme solution. . As shown in FIG. 1, the neutral protease increased in activity from the beginning of fermentation, showing maximum activity at 48 hours, and then decreased in activity. The acidic protease was slightly lower in activity than in the early stage of fermentation and then increased in activity, showing maximum activity at 48 hours, but then decreased. Alkaline protease, like acidic protease, decreased in activity at the beginning of fermentation, then increased to show maximum activity at 36 hours, and then decreased, but was generally less active than acidic or neutral protease.

Experimental Example  4. Optimum pH and Temperature of Protease

10 g of Cheonggukjang meju, which was incubated for a certain period of time after the addition of the Bacillus licheniformis strain, was taken in a Erlenmeyer flask. pH 9.0), 25 mM borax-NaOH buffer (pH 10.0) was added at 100 mL each, shake-extracted at room temperature for 24 hours, and the solution was filtered to measure the protease activity of the filtrate to determine the optimal pH for proteolytic elution. It was. In the preparation of the present invention Cheonggukjang after adding Bacillus richeniformis strain after incubation for a certain time in a constant temperature of 20 ℃, 30 ℃, 35 ℃, 40 ℃, 50 ℃ and 60 ℃, respectively, by adding a buffer of the predetermined pH to extract the shaking After filtration, the protease activity of the filtrate was measured to determine the optimum temperature.

As a result of measuring the relative activity of the enzyme according to pH among the factors influencing the degree of decomposition elution of the protein during the fermentation process. The optimum pH of the protease in Cheonggukjang fermentation with added Bacillus Richenomyces strain showed the highest degradation rate at 6.5.

Figure 3 shows the results of measuring the relative activity of the enzyme when reacted with varying the temperature in order to obtain the optimal temperature of the proteolytic enzyme during the fermentation process of Cheonggukjang added Bacillus licheniformis strain. It gradually increased with increasing temperature and showed the highest dissolution rate at 35 ° C., but its activity decreased with increasing temperature.

Experimental Example  5. Protein  Separation and Quantification

Cheonggukjang added Bacillus licheniformis strain of the present invention was separated from the water-soluble protein and the salt-soluble protein according to Wang's classification method, and the quantification of the protein was measured by Lowry et al.

The contents of the water-soluble protein and the salt-soluble protein are shown in Table 3. As the fermentation time passed, the contents of water-soluble protein and salt-soluble protein tended to increase.

Change in Protein Content during Fermentation of Cheonggukjang of the Present Invention (mg / g) division Fermentation period (hours) 0 6 12 24 36 48 60 Water soluble protein 6.48 7.53 8.14 8.88 9.92 10.82 12.23 Soluble Protein 5.20 6.48 7.21 8.12 9.41 11.12 11.21

Experimental Example  6. Determination of Molecular Weight of Main Protein

Molecular weight of main protein was measured by sodium dodecyl sulfate polyacrylamide gel electrophoresis and developed according to the method of Weber and Osborn. The molecular weight was measured using standard curve according to Rm value. As standard, bovine serum albumin (MW34,700), egg albumin (egg albumin, MW45,000), pepsin (pepsine, MW34,700), trypsinogen (trypsinogen (MW24,000), beta-lactoglobulin (β- lactoglobulin, MW18,400), lysozyme (MW14,300) were used.

As a result of measuring the molecular weight of the main protein during the fermentation time of the Cheonggukjang added Bacillus lynchiniformis strain of the present invention, the molecular weight of the water-soluble protein at 48 hours of fermentation was about 19,000 (see FIG. 4).

Experimental Example  7. Analysis of changes in amino acid composition

Amino acid analysis of water soluble and soluble protein was performed by placing 20 mg of each protein in a 5 mL glass tube and hydrolyzing with 6 M HCl for 24 hours. The hydrolyzed digest was filtered and hydrochloric acid was completely evaporated using a rotary evaporator at 40 ° C. or lower, and then dissolved in 10 mL sodium citrate buffer (pH 7.0).

Table 4 shows the changes in amino acid composition of water-soluble protein during the fermentation process of Cheonggukjang added Bacillus Richenomyces strain of the present invention. Immediately after the increase of the Bacillus licheniformis strain, a total of 16 amino acids of the water-soluble protein, proline was the highest, followed by glutamic acid and serine. Serine, proline and histidine decreased at 24 hours of fermentation, and glutamic acid, aspartic acid and phenylalanine at 60 hours of final fermentation. Its content was high. The total amino acid content reached the maximum value of 102.22 mg / g at 24 hours of fermentation, and there was no significant difference in the change of total amino acid content during the whole fermentation process.

Analysis of Amino Acid Content of Water Soluble Protein during Fermentation of Cheonggukjang of the Present Invention (mg / g) Type of amino acid Fermentation period (hours) 0 24 48 60 Aspartic acid 7.53 11.80 9.28 12.78 Threonine 2.62 4.47 4.77 4.80 Serine 8.41 4.57 4.89 4.60 Glutamic acid 12.61 18.21 13.14 13.18 Prorin 21.41 10.03 4.61 3.25 Glycine 3.21 4.71 5.35 6.26 Alanine 2.95 4.17 6.29 5.72 Cystine Not detected Not detected Not detected Not detected Varin 2.49 5.12 5.12 4.97 Methionine 0.63 0.80 1.27 1.54 Isoleucine 2.66 4.83 4.57 3.81 Leucine 3.82 6.10 7.81 4.18 Tyrosine 1.41 3.48 5.14 9.24 Phenylalanine 2.56 5.86 8.43 9.03 Histidine 5.24 5.00 6.01 8.29 Lysine 5.12 5.94 6.30 7.01 Argin 5.19 7.13 5.15 5.13

The amino acid composition of the salt-soluble protein during fermentation process of Cheonggukjang added Bacillus licheniformis strain of the present invention is shown in Table 5. A total of 16 amino acids of soluble protein were identified, followed by phenylalanine, followed by glutamic acid and aspartic acid. Like the amino acid composition of water-soluble protein, cystine was not detected during the fermentation process, and very little methionine was contained at the beginning of fermentation, which is thought to be destroyed by acid decomposition.

Analysis of Amino Acid Content of Salt Soluble Protein during Fermentation of Cheonggukjang of the Present Invention (mg / g) Type of amino acid Fermentation period (hours) 0 24 48 60 Aspartic acid 11.42 4.71 5.63 5.21 Threonine 6.30 2.33 3.44 4.03 Serine 5.09 3.89 3.89 3.72 Glutamic acid 24.12 8.56 8.70 8.85 Prorin 7.94 3.14 2.97 2.35 Glycine 6.28 2.58 3.49 4.26 Alanine 5.62 2.22 3.05 3.07 Cystine Not detected Not detected Not detected Not detected Varin 3.88 2.18 2.60 2.54 Methionine 0.16 1.18 0.98 0.83 Isoleucine 3.41 2.24 2.38 2.16 Leucine 5.93 3.31 3.44 3.37 Tyrosine 3.37 2.14 1.84 2.62 Phenylalanine 4.24 2.87 4.03 4.17 Histidine 5.82 5.45 4.97 4.38 Lysine 9.21 3.88 3.92 3.16 Argin 8.63 2.50 3.65 3.28

As described above, according to the present invention, Bacillus licheniformis strain may be added in the preparation of Cheonggukjang so that the content of cystine, which is a water-soluble amino acid related to Cheonggukjang inherent odor, may be improved, thereby providing a new Cheonggukjang with improved flavor. It is an excellent invention in the food industry because it has an excellent effect.

1 is a graph showing the changes of proteolytic enzymes during the fermentation period of Chunggukjang of the present invention.

Figure 2 is a graph showing the activity of the protease according to pH during the fermentation process of Cheonggukjang of the present invention.

Figure 3 is a graph showing the activity of the protease according to the temperature during the fermentation process of Cheonggukjang of the present invention.

Figure 4 is a graph showing the molecular weight measurement of the main protein during the fermentation process of Cheonggukjang of the present invention.

Claims (1)

Soybeans were soaked in 20 ℃ water for 15 hours, dehydrated for 1 hour, steamed for 1 hour, cooled to 50 ℃, inoculated with Bacillus licheniformis CY-1480 (KACC 91325P) strain, and left for 36 ~ 60 hours at 35 ~ 38 ℃. The main protein molecular weight (MW) is 19,000, characterized in that the production of Cheonggukjang by culturing, the water-soluble amino acid cystine (cystine) associated with the smell is lost, the production method of Cheonggukjang improved.

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