KR100590913B1 - A method for preparation of chungkukgang with soybean curd residue by using bacillus sp. and lactic acid bacteria - Google Patents

Abstract

The present invention relates to a method for preparing Cheonggukjang with Beechi using Bacillus subtilis and Lactic Acid Bacteria, by adding battery-activated live soybean fine powder to Beechi using a Bacillus subtilis to obtain a one-stage fermentation product containing viscous substance and thrombolytic enzyme. By adding lactic acid bacteria and skim milk to the first stage fermentation, two stages of fermentation enhances protein, improves flavor, increases peptide content, and improves the odor of peculiar fermentation. In other words, there is an excellent effect to provide the Cheonggukjang.

B., Bacillus subtilis, Lactic Acid Bacteria, Cheongguk Fermentation, Lactic Acid Fermentation, Whole Active Soybean Fine Powder, Bacillus firmus NA, Bacillus subtilis GT, Lactobacillus casei LS

Description

A method for preparation of Chungkukgang with soybean curd residue by using Bacillus sp. and lactic acid bacteria}

Figure 1 is a process diagram schematically showing the process of fermented soybeans.

Figure 2 is a graph showing the peptide content after one-stage fermentation by Bacillus pilmus and Bacillus subtilis by using 20% by weight of the active and soybean fine powder added soybean fine powder. Where * indicates the addition of 20% by weight of the whole active soybean fine powder, 0% is the fermentation product fermented only with Bacillus pilmus, JAP ^* is the fermentation product of Bacillus pilmus and whole active soybean fine powder, GT-D ^* Denotes a fermentation product of Bacillus subtilis and whole active soybean fine powder.

FIG. 3 is a graph showing the thrombolytic enzyme activity of fermented products by one-stage fermentation using Bacillus pilmmus and Bacillus subtilis of 20% by weight of whole-active soybean fine powder and non-added biji. * Indicates that 20% of the whole active soybean fine powder is added, 0% is the fermentation product fermented only with Bacillus pilmus, JAP ^* is the fermentation product of Bacillus pilmus and whole active soybean fine powder, GT-D ^* Represents a fermentation product of Bacillus subtilis and whole active soybean fine powder.

Figure 4 is a graph measuring the viscosity of the fermentation product by one-stage fermentation using Bacillus pilmus and Bacillus subtilis of the non-added and non-added biji with 20% by weight of all-active soybean fine powder. Where * indicates the addition of 20% by weight of the whole active soybean fine powder, 0% is the fermentation product fermented only with Bacillus pilmus, JAP ^* is the fermentation product of Bacillus pilmus and whole active soybean fine powder, GT-D ^* Denotes a fermentation product of Bacillus subtilis and whole active soybean fine powder.

FIG. 5 is a graph showing the peptide content of fermentation product after fermentation by Bacillus filmus and Bacillus subtilis and 2-stage fermentation with Lactobacillus cascai of 20 wt% of all-active soybean fine powder. to be.

FIG. 6 shows the thrombolytic enzyme activity of fermented product after fermentation by Bacillus filmus and Bacillus subtilis and 2-stage fermentation using Lactobacillus cascai of 20 mg by weight of battery-activated raw soybean fine powder. The graph shown.

The present invention relates to a method for producing Cheonggukjang with Beechi using Bacillus subtilis and Lactic Acid Bacteria, and more specifically, Bacillus firmus and Baciilus subtilis Bacillus subtilis 1 However, after performing fermentation, two-stage fermentation, which is a mixed fermentation form with Lactobacillus cas ei LS, is performed, and the flavor is excellent, and a functional busy fermentation product containing a viscous substance, a peptide, a thrombolytic enzyme, ie, cheonggukjang is prepared. It is about how to.

Soybean is used in various traditional foods such as soybeans and bean sprouts, and as a primary processed food of soybeans, high quality tofu has been used as an important protein source of Asians as a protein concentrate.

Tofu production has traditionally been concentrated and pressed soy protein, soy whey and bean curd as by-products. In particular, it is a material that contains nutrients of soybeans, and is a material that is easily deteriorated by microorganisms contamination and microorganisms present in the tofu manufacturing process and the drying process of the bean curd, despite the high fiber content. Therefore, the utilization of busy papers is an important research task not only in terms of the use of by-products, but also in terms of reducing the treatment cost of environmental pollutants.

Until now, the study on busy fat has been studied mainly on drying of busy fat in order to extend the shelf life until it is processed. Is inadequate.

The present inventors added two kinds of strains, Bacillus pilmus and Bacillus subtilis, isolated from Cheonggukjang, to fermented biscuits containing whole-active raw soybean fine powder, followed by fermentation of Bacillus subtilis, followed by addition of whole-active soybean fine powder. By confirming the effect of the reinforcement of protein and increase of the peptide, thrombolytic enzyme, viscous product production and flavor enhancement, and the final fermented fermented product by performing mixed fermentation using lactic acid bacteria, Lactobacillus cascai to the fermented fermentation product The present invention has been completed by producing a fermented product, ie, Cheonggukjang, containing bioactive substances including lactic acid bacteria together with Bacillus subtilis as a probiotic while reducing the peculiar smell of Cheonggukjang compared to the conventional one-stage fermentation.

Therefore, an object of the present invention is to provide a method for producing Cheonggukjang by using a Bacillus subtilis and lactic acid bacteria.

The object of the invention is the following Bacillus peel mousse (Bacillus firmus NA) and Bacillus subtilis (Bacillus subtilis GT) having a busy addition of the cell active raw soybean fine powder sterilized for 15 minutes at 121 ℃ using an autoclave probe It was achieved by inoculating 1% each and fermenting at 42 ° C. for 18 hours, followed by fermentation at 37 ° C. for 16 hours by adding 1% of Lactobacillus casei LS and 10% of 10% skim-milk solution. .

Hereinafter, the configuration and operation of the present invention.

The present invention is a step of sterilizing 10 ~ 30 minutes at 110 ~ 130 ℃ after adding 10 ~ 30% by weight of battery-activated raw soybean fine powder to the busy; Bacillus firmus and Bacillus subtilis culture solution of the mixture of the above step was inoculated 0.1 to 3% by weight, respectively, and then fermented at 37 to 45 ℃ for 15 to 25 hours to fermentate the first stage. Obtaining; And Lactobacillus casei LS ( Lactobacillus casei LS) to the fermentation step of 1 to 5% by weight and 10% skim milk 5-30% by weight of the two-stage fermentation step.

If the addition amount of Bacillus pilmus, Bacillus subtilis and Lactobacillus casei added during fermentation and fermentation time in the present invention, and the fermentation time is outside the above range, the flavor reduction of the fermentation product obtained after fermentation, the viscosity content and the effect of increasing the production of thrombolytic enzyme In addition, there is a disadvantage in producing the cheonggukjang with increased ammonia smell.

The most preferred method for producing Chunggukjang of the present invention is the step of sterilizing for 15 minutes at 121 ℃ by adding 20% by weight of the battery-activated raw soybean fine powder to the busy; Bacillus firmus ( Bacillus firmus ) and Bacillus subtilis ( Bacillus subtilis ) inoculated by 1% by weight each of the raw material-treated fermentation at 18 ℃ for 18 hours to obtain a single-stage fermentation; And adding 1% by weight of Lactobacillus casei LS and 10% by weight of 10% skim milk to the first stage fermentation.

Bacillus firmus ( Bacillus firmus ) and Bacillus subtilis ( Bacillus subtilis ) used in the present invention can be used as long as any strain belonging to the same species (sepecies) separable from Cheonggukjang, lactobacillus casei (lactic acid bacterium) Lactobacillus casei ) can be used if the strain belongs to the same species (species).

Hereinafter, the configuration of the present invention will be described in more detail with reference to Examples, but the scope of the present invention is not limited only to the following Examples.

Example 1 fermented product of the present invention bean curd, Cheonggukjang

First step: raw material processing

20% by weight of battery-activated raw soybean fine powder was added to the sebum and then sterilized at 121 ° C. for 15 minutes by autoclave.

Second step: preparing single-stage fermented biji with Bacillus

In order to ferment the Cheonggukjang to the raw material prepared in the first step, the starters of Bacillus firmus NA ATCC8247 and Bacillus subtilis GT-D AY583216, 1 wt% Inoculated and fermented at 42 ° C. for 18 hours.

Bacillus starter was produced by Bacillus firmus NA ATCC8247 and Bacillus subtilis GT-D AY583216 separated by Cheonggukjang for 24 hours using a 42 ° C thermostat. Then, one colony (colony) was inoculated in sterilized 5% soybean powder solution and then shake cultured at 150 rpm for 24 hours in a 42 ℃ thermostat was used as a starter spawn.

Third step: production of two-stage fermented biji using lactic acid bacteria

Lactobacillus casei LS 1% by weight and 10% by weight of 10% skim milk were added to the fermented Chungkookjang fermentation in the second step, followed by incubation for 16 hours at 37 ° C. in a two-stage fermentation. A fermentation product was prepared.

The lactobacillus starter was prepared by activating Lactobacillus casei LS ATCC393 on an MRS agar plate containing MRS for 24 hours at 37 ° C., followed by soybean powder solution containing sterilized skim milk. Scheme milk: soybean powder = 1: 4) and then incubated for 24 hours in a 37 ℃ thermostat was used as starter spawn.

Experimental Example 1: Investigation of Physicochemical Properties Differences in Busy Fermented with or without Whole-Active Raw Soybean Fine Powder

Figure 2 is a graph showing the peptide content of fermented products with or without addition of battery-activated raw soybean fine powder when single-stage fermentation using Bacillus pilmus and Bacillus subtilis according to the present invention, Figure 5 is also the same method It is a graph showing the peptide content of the fermented product when two-stage fermentation is performed using Lactobacillus casei and skim milk after one-stage fermentation.

Peptide content of the busy fermentation product prepared in Example 1 was mixed with 80 mL distilled water to 20 g busy fermentation, centrifuged (15000rpm, 15 minutes), the supernatant was taken and quantified by total phenol (total phenol) measurement method.

It was found that the peptide content was increased in the added fat compared to the non-battery-free soybean powder, and the peptide content was increased in the two-stage fermentation than in the one-stage fermentation. It was considered that fermentation of Bacillus subtilis and Lactic acid bacteria occurred during the time.

Moisture and Crude Protein Content (%) of Bacillus firmus Fermentation of Baji using Whole Active Soybean Fine Powder division A ^* B ^** Water content (%) 84.2 69.4 Crude Protein (%) 6.9 28.4 1. A *: Before adding whole active soybean fine powder 2. B **: After adding whole active soybean fine powder

As shown in Table 1 above, by adding 20% of all active soybean fine powder, the water content of 84.2% of the sebum was reduced to 69.4% and 15% of water content. Could see.

In addition, when not added, the inherent odor and viscosity of Cheonggukjang were not produced, but after the addition, a sweet smell, which is peculiar to Dannae and Cheonggukjang, was also produced. Therefore, the addition of battery-activated raw soybean fine powder in non-fermentation greatly contributed to the improvement of quality and the production of functional substances of fermented products.

Experimental Example 2: Investigation of the physical properties of busy fermented products with or without whole-active soybean fine powder

)는 1~100 s^-1의 범위로 유동특성을 알아보았고, 점조도지수와 유동지수 값은 파워 로우 모델(Power law model)(τ=ar^b)로 측정하였다.The viscosity of the busy fermentation product was measured using a measuring plate P61 by mounting a spindle (Platte PP35 Ti, D = 35 mm) on a rheometer system (HAAKE RheoStress 1, Germany). After adding 80 mL of distilled water to 20 g of fermentation, 1 mL of sample was taken, and the average value was measured for 10 seconds per section. The value obtained was expressed as shear rate (1 / s) and shear stress (Pa). As the height was increased, the effect of increasing the laminar strain was measured. Shear speed at 20 ℃

The flow characteristics were measured in the range of 1 to 100 s ^-1 , and the viscosity index and the flow index were measured by the power law model (τ = ar ^b ).

As a result of the experiment, when 20 wt% of the battery-activated raw soybean fine powder was added as shown in Table 2, the a value representing the consistency was increased by about 4 times, and fermented by Bacillus pilmus rather than Bacillus subtilis as shown in FIG. 4. It was found that the busy had a higher viscosity (viscosity).

Differences in Viscosity of Bacillus subtilis Fermented with Bacterium-Activated Raw Soybean Fine Powder Total activity soybean powder concentration (%) Power law index a b R ² Bacillus Pilmus 0 0.05 0.45 0.9728 20 0.2469 0.8917 0.9926 Bacillus subtilis 20 0.09 0.759 0.9999 1. τ = ar ^b a: Fluid consistency coefficient, b: Flow behavior index of fluid, R ² : Regression coefficient

Experimental Example 3: Investigation of viable cell counts and thrombolytic enzyme activity of fermented product prepared with BJ added with whole active soybean fine powder

Figure 3 shows the thrombolytic enzyme activity of the fermentation product by one-stage fermentation according to the present invention when the one-stage fermentation using Bacillus pilmus and Bacillus subtilis according to the present invention. 6 shows the thrombolytic enzyme activity of the fermented product by two-stage fermentation using Lactobacillus casei and skim milk after one-stage fermentation.

The measurement of the viable cell count of the fermented fermentation product prepared by the present invention was performed by mixing 80 mL of sterile distilled water with 20 g of the fermented fermentation product, diluting it with sterile distilled water, and then stating the plate on an MRS agar plate containing MRS. The number of colonies shown after incubation for 24 hours in a room temperature was measured.

Thrombolytic enzyme activity was measured using the Astrup and Mullertz method, a kind of fibrin plate method. Fibrin plates were dissolved in 0.5% fibrinogen in 0.067M sodium phosphate buffer (pH 7.4) and 10 mL was added to a petri dish 9 cm in diameter. 0.1 ml of thrombin (100 NIH / mL) dissolved in 0.067 M sodium phosphate buffer was added thereto, mixed quickly, and a uniform plate was prepared and used for 30 minutes at room temperature.

Sample drop position was indicated on a fibrin plate, and 20 µl each of the fermentation extracts was dropped at the indicated position for 2 hours at 37 ° C. incubator, and the enzyme activity was determined by dissolution area. To determine the standard curve of thrombolytic enzymes, standard plasmin solution was prepared in Tris-lysine buffer (pH 9.0) to 0.6, 1.6, 2.6, 5 units / mL, and then fibrin plates. A standard curve of plasmin enzyme activity was prepared from the dissolved area formed by adding 20 µl to the fibrin plate. The thrombolytic enzymes in the fermented product were converted into plasmin units compared to the standard curve, and the thrombolytic enzyme activity was determined by the following method.

Thrombolytic activity (%) = (sample lysis zone / plasmin lysis zone) × 100

As a result, it was found to be about 3 times higher in the biji which was added with all-active raw soybean fine powder. Bacillus Philmus (B. firmus) Bacillus subtilis (B. subtilis)).

As a result of the measurement of thrombolytic enzymes, the fermented biji fermented with Bacillus pilmus showed high thrombolytic activity, and the thrombolytic activity of each strain was decreased in two strains.

Number of live cells (cfu / g) fermented from biji with added active soybean fine powder 1st stage fermentation Two-stage fermentation Busy Fermentation Using Bacillus Pilmus 5 × 10 ⁹ 5.5 × 10 ⁹ * 3 × 10 ⁹ Busy Fermentation Using Bacillus Subtilis 4.5 × 10 ⁹ 2.5 × 10 ⁹ * 4 × 10 ⁸ 1. *: Number of live bacteria ( Lactobacillus casei LS)

As a result of measuring the viable cell count, Bacillus fermented fermented with two kinds of Bacillus was able to confirm the number of viable bacteria of 10 ⁹ at the first stage fermentation, and lactic acid bacteria at 4 × 10 ^{8-10 9} Viable cell counts could be seen together.

As described above through the above Examples and Experimental Examples, the present invention is to obtain a one-stage fermentation product containing viscous substance and thrombolytic enzyme using Bacillus subtilis by adding battery-activated raw soybean micropowder to the biji and the one-stage fermentation product. By adding lactic acid bacteria and skim milk to the two-stage fermentation, the protein is strengthened, the flavor is improved, the peptide content is increased, the peptide content is increased, and the viable fermentation product can reduce the odor peculiar to Cheonggukjang by virtue of the high lactic acid bacteria viable count and surviving together In other words, because it has an excellent effect of providing the soybean soup is a very useful invention in the food industry.

Claims (2)

Adding 10-30% by weight of battery-activated raw soybean fine powder to the sebum and sterilizing at 110-130 ° C. for 10-20 minutes; Bacillus firmus and Bacillus subtilis culture solution of the mixture of the above step was inoculated 0.1 to 3% by weight, respectively, and then fermented at 37 to 45 ℃ for 15 to 25 hours to fermentate the first stage. Obtaining; And Lactobacillus casei LS ( Lactobacillus casei LS) 0.1 to 5% by weight and 10 to 30% skim milk by adding to the fermentation of the first stage fermentation method comprising the step of fermentation . Cheonggukjang made from bean cured by the method of claim 1.

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