KR101740147B1 - Method of reducing biogenic amines - Google Patents

Abstract

The method for reducing biorenic amines according to the present invention comprises the steps of: cultivating Bacillus subtilis in a culture medium having a salt concentration of 1 to 5%, culturing the mixture at a temperature of 30 to 35 DEG C at 200 rpm for 12 hours with shaking; A second step of preparing a mixed solution of sodium alginate by adding the above-mentioned cultured Bacillus subtilis to 1 to 4% sodium alginate solution; A third step of dropping the sodium alginate mixed solution into a calcium chloride solution having a concentration of 0.5 to 3 moles to prepare beads; A fourth step of curing the beads in a calcium salt solution for 30 minutes to form an immobilized bacterium carrier; A fifth step of introducing a salt solution having a salt concentration of 5 to 20% into the reactor filled with the immobilized Bacillus subtilis carrier of the fourth step and activating the immobilized Bacillus subtilis through a salt solution adaptation period for 8 to 12 days; A sixth step of adapting the immobilized Bacillus subtilis activated through the salt solution adaptation period of the fifth step to the fish sauce for 1 to 2 weeks; The immobilized Bacillus subtilis adapted to the sixth step is supplied to the fish sauce to decompose the amines, and the immobilized Bacillus subtilis adapted to the fish sauce is circulated in the reactor every 12 to 24 hours by using a peristaltic pump to produce a biogenic amine component And a seventh step of reacting the reaction mixture for 1 to 3 months.
In addition, the culture liquid includes yeast concentrate, mannitol, salt, ammonium phosphate, magnesium sulfate, and distilled water, and is characterized by a ratio of 1 to 10% based on the total weight of the raw materials for manufacturing the fermented fish meal.
The volume ratio of the immobilized Bacillus subtilis carrier packed in the reactor in the fifth step is 10 to 20%.

Description

Method of reducing biogenic amines [0002]

The present invention relates to a method for reducing biorenic amines, and more particularly, to a method for removing biogenic amine components produced during fermentation and fermentation of fermented fish sauce or fish sauce using immobilized Bacillus subtilis.

It is a fish fermented food which salted seafood and digested by self-digesting enzymes or microbial enzymes to give unique flavor. According to recent data, there are more than 100 varieties of salted fish. The amount of fermented seafood produced in Korea is 27,228 tons (2011), which is officially confirmed. The highest production of fermented seaweeds is 10,999 tons. It is estimated that the per capita intake of seafood per adult per year is 1 kg per year, indicating that Korea belongs to the culture of salted fish.

The problem of salted fish is very weak in sanitary management by small self - processing of small - size seafood which is low quality in the case of traditional salted fish. For example, there is a problem in that the control of the temperature of the environment surrounding the facility is not well managed, such as using raw materials that are not well managed after the catch, or controlling the propagation of the early corrupt microorganisms. When the fish oil protein components of the fermented fish are fermented and aged, the biogenic amine component is excessively produced due to the decarboxylation by the microorganisms.

The biogenic amine component acts on the central nervous system and the like in the human body and causes food poisoning allergy. Histamine, the most widely known of biogenic amines, causes food poisoning with atopic and vomiting, diarrhea, stomach pain, vascular effects (hypertension, heart rate), headache, kidney poisoning, dyspnea, itching, rash and fever. Tyramine acts on the vasoconstriction and can lead to fatal hypertension, such as a cheese reaction, and migraine. We have enzymes that degrade biogenic amines in the small intestine. However, if you drink too much, drink drugs, or have a small intestine disease, even if you take a small amount of biogenic amine, harmful symptoms may appear on your body.

Biogenic amines are also included in fermented foods known to be good for our bodies such as miso, soy sauce, chonggukjang, traditional fermented beverages, and wine. The KFDA has proposed a method of applying low temperature fermentation (below 30 ℃) and low temperature circulation (below 4 ℃) to reduce the biogenic amine content. However, due to the characteristics of fermented foods such as soybean paste, soy sauce, and fermented fish meal, fermentation and aging processes are essential for long-term storage, so that it is difficult to achieve reduction of biogenic amine components in a short time. In particular, in the case of fish sauce, the fermentation process is performed for 1 to 2 years, so that the reduction of histamine is more difficult. Most of the commercially available fish sauces have histamine content over 1000ppm.

Germany, France and Europe have a recommended standard of histamine for wine. The International Commission on Codex Alimentarios (CODEX) defines biosynthetic amines as 100 to 200 mg per kilogram for fish and processed fish. However, in Korea, fermented foods are traditionally enjoyed, but standards and specifications for biogenic amines are not available.

Biogenic amines, which are present in large quantities in fermented foods such as salted fish, are harmful to human body and cause allergy or food poisoning.

Korean Patent Publication No. 10-2012-0092230

It is an object of the present invention to provide a high-quality salted and fermented fish sauce by removing the biogenic amine component produced during fermentation and fermentation of fermented fish sauce or fish sauce using immobilized Bacillus subtilis.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention as set forth in the accompanying drawings. It will be possible.

The method for reducing biorenic amines according to the present invention comprises the steps of: cultivating Bacillus subtilis in a culture medium having a salt concentration of 1 to 5%, culturing the mixture at a temperature of 30 to 35 DEG C at 200 rpm for 12 hours with shaking; A second step of preparing a mixed solution of sodium alginate by adding the above-mentioned cultured Bacillus subtilis to 1 to 4% sodium alginate solution; A third step of dropping the sodium alginate mixed solution into a calcium chloride solution having a concentration of 0.5 to 3 moles to prepare beads; A fourth step of curing the beads in a calcium salt solution for 30 minutes to form an immobilized bacterium carrier; A fifth step of introducing a salt solution having a salt concentration of 5 to 20% into the reactor filled with the immobilized Bacillus subtilis carrier of the fourth step and activating the immobilized Bacillus subtilis through a salt solution adaptation period for 8 to 12 days; A sixth step of adapting the immobilized Bacillus subtilis activated through the salt solution adaptation period of the fifth step to the fish sauce for 1 to 2 weeks; The immobilized Bacillus subtilis adapted to the sixth step is supplied to the fish sauce to decompose the amines, and the immobilized Bacillus subtilis adapted to the fish sauce is circulated in the reactor every 12 to 24 hours by using a peristaltic pump to produce a biogenic amine component And a seventh step of reacting the reaction mixture for 1 to 3 months.
In addition, the culture liquid includes yeast concentrate, mannitol, salt, ammonium phosphate, magnesium sulfate, and distilled water, and is characterized by a ratio of 1 to 10% based on the total weight of the raw materials for manufacturing the fermented fish meal.
The volume ratio of the immobilized Bacillus subtilis carrier packed in the reactor in the fifth step is 10 to 20%.

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According to the solution of the above-mentioned problem, the method of the present invention for reducing the bio-genic amines can be used to produce high quality pickles and fish sauces by removing the biogenic amine components produced during the fermentation and fermentation of the fermented fish paste or the fish sauce using immobilized Bacillus subtilis It is effective.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flowchart showing a method for reducing a biogenic amine according to the present invention. FIG.

The above and other objects, features and advantages of the present invention will be more apparent from the following detailed description taken in conjunction with the accompanying drawings, in which: FIG. BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention and the manner of achieving them will become apparent by reference to an embodiment which will be described in detail below with reference to the accompanying drawings.

Hereinafter, the method for reducing the biogenic amine described above will be described in detail with reference to the drawings.

<Method for reducing biogenic amine>

First, in the first step (S1), Bacillus subtilis is cultured in a culture medium. Concretely, Bacillus subtilis is cultured in a culture medium having a salt concentration of 1 to 5%. At this time, the culture liquid is used in a proportion of 1 to 10% based on the total weight of the raw materials for manufacturing the fermented fish meal.

If the culture medium is used at a concentration lower than 1%, culture may not be effectively performed. If the culture medium is used at a concentration higher than 10%, the economic efficiency is lowered and there is no significant difference in the culture rate of Bacillus subtilis. Therefore, the culture medium is used at a ratio of 1 to 10% .

The Bacillus subtilis is cultivated in a culture medium comprising 0.5 to 2% of yeast concentrate, 0.5 to 3% of mannitol, 0.1 to 5% of salt, 0.1 to 2% of ammonium phosphate, 0.01 to 0.2% of magnesium sulfate and distilled water, And cultured under shaking at 200 rpm for 12 hours.

The step of culturing the Bacillus subtilis may be applied to the production process of fermented fish products such as fermented fish products or fish sauces using small-sized fishery products, thereby removing the biogenic amines contained in the fermented fish sauce or fish sauce.

The biogenic amine is a food poisoning-inducing component contained in a fermented fish sauce or a fish sauce, and is produced by fermentation and aging of a fish protein component of a fermented fishery product, resulting from decarboxylation by microorganisms.

Next, in the second step (S2), a sodium alginate mixed solution is prepared. Specifically, the cultured Bacillus subtilis is added to the above 1 to 4% sodium alginate solution at a ratio of 2 to 4%, and the mixture is mixed and stirred to prepare a sodium alginate mixed solution.
Next, in a third step (S3), a bead is produced. Specifically, the prepared sodium alginate mixed solution is allowed to stand for 10 minutes. Thereafter, the prepared sodium alginate mixed solution is dropped in small amounts to a calcium chloride solution having a concentration of 0.5 to 3 mol, to prepare a transparent bead.
The sodium alginate solution is derived from natural seaweed, and has an excellent food safety. If the concentration of the sodium alginate solution is lower than 1%, the gel is not formed well and it is difficult to produce the beads. If the concentration of the sodium alginate solution is higher than 4%, sodium alginate is not dissolved well and the viscosity is high. There is a difficult problem.
Next, in the fourth step (S4), the immobilized bacterium carrier is formed. Specifically, the beads are cured in a calcium salt solution for 30 minutes to form an immobilized bacterium carrier. The immobilized Bacillus subtilis carrier that has been formed is washed with distilled water three times or more. The size of the immobilized bacterium carrier is 3 to 5 mm, and is determined by the viscosity of the mixed sodium alginate solution and the size of the discharge port of the dispenser.

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Next, in the fifth step (S5), the immobilized Bacillus subtilis is activated. Specifically, a salt solution in which salt is dissolved is introduced into the reactor filled with the immobilized Bacillus subtilis carrier to activate immobilized Bacillus subtilis.

The reactor is prepared by placing the immobilized Bacillus subtilis carrier in a cylindrical tube, filling the tube with purified water or physiological saline, and sealing both ends of the tube.

The immobilized Bacillus subtilis carrier is filled in the reactor. At this time, the volume ratio of the immobilized Bacillus subcomponent is 10 to 20%, and the immobilized Bacillus subtilis carrier in the reactor is circulated by the peristaltic pump. Also, the flow rate was adjusted so that the immobilized bacteria in the reactor could stay for 10 to 15 hours, and the salt concentration of the salt solution was increased to 25% so as to be activated in the salt solution, Adjust to adapt. In addition, the immobilized Bactobacillus carrier adsorbs microorganisms in the reactor and reacts with the amine component more effectively to accelerate the decomposition of the amine.

If the volume ratio of the immobilized Bacillus subtilis carrier is less than 10%, the removal rate of harmful substances by the immobilized Bacillus subtilis is low, which is not efficient. If the volume ratio of the immobilized Bacillus subtilary carrier is higher than 20%, economical efficiency is deteriorated.

Next, in the sixth step (S6), the activated immobilized Bacillus subtilis is adapted to the salted or fermented fish sauce. Specifically, immobilized Bacillus subtilis immobilized in the salt solution is added to the fish sauce, and the fish sauce is adapted to the fish sauce for 1 to 2 weeks.

Next, in a seventh step (S7), the immobilized Bacillus subtilis adapted to the fermented fish or fish sauce is used to decompose the amine. Specifically, immobilized Bacillus subtilis adapted to a salted or fermented fish sauce is reacted with a biogenic amine component contained in a fish sauce using a peristaltic pump in a reactor.

In the reactor, immobilized Bacillus subtilis is circulated using a peristaltic pump every 12 to 24 hours, and the amine component is decomposed by reacting with the biogenic amine component contained in the fermented fish sauce or fish sauce for 1 to 3 months.

The immobilized Bacillus subtilis adapted to the fermented salted and fermented fish sauce has the characteristic of decomposing the biogenic amine component which is the food poisoning substance produced during the fermentation of fermented seaweed or fish sauce having an average salinity of 20 to 25%, thereby reducing the allergic histamine It is effective.

When the fermented Bacillus subtilis is used to produce the fermented fish sauce and the fermented fish sauce, high quality fermented fish sauce and fish sauce having few harmful substances such as histamine can be produced.

Hereinafter, experimental data for examining the change of the number of immobilized Bacillus subtilis according to the reaction time in the method of reducing the biogenic amine in the present invention will be described in detail.

Changes in number of immobilized Bacillus subtilis according to reaction time

In the following experiment, in order to examine the change of the number of immobilized Bacillus subtilis according to the reaction time in the method of reducing the biogenic amine in the present invention, the number of immobilized Bacillus subtilis was set to 2.1 × 10 ² The culture was filled and a 5% saline solution was introduced. Then, the inside of the reactor packed with alginic acid beads was adjusted to circulate every 12 hours using a peristaltic pump, and the change of immobilized Bacillus subtilis over time was measured.

Reaction time (days) 10 20 30 40 60 Viable cell count 3.6 x 10 ³ 3.9 × 10 ⁴ 10.1 × 10 ⁴ 2.6 x 10 ³ 5.5 × 10 ³

As shown in Table 1, it can be seen that the number of bacteria tends to increase as the reaction time increases.

In addition, the number of bacteria increased gradually, and the number of immobilized Bacillus subtilis in the culture medium remained constant and stabilized during the reaction period after 30 days of reaction time.

Hereinafter, in order to examine the effect of removing the histamine component of the immobilized Bacillus subtilis in the method of reducing the biogenic amine according to the present invention, the experimental contents in which immobilized Bacillus subtilis was filled in the reactor and reacted with the anchovy sauce was described in detail.

The concentration of histamine in anchovy sauce according to the presence of immobilized Bacillus subtilis

The following experiment was conducted to compare histamine concentrations of anchovy sauce according to whether the immobilized Bacillus subtilis was added or not. The histamine content of the anchovy sauces prepared by the same method was measured by varying the amount of immobilized Bacillus subtilis.

[Comparative Example 1]

Comparative Example 1 is an anchovy sauce prepared by adding sun salt to an anchovy at an ammonium concentration of 23% by weight relative to the weight of the anchovy, mixing the anchovy and salt well, and aging and fermenting at room temperature for 10 months.

[Example 1]

In Example 1, 20 kg of anchovy was added with salt of salt at a salt concentration of 23%, anchovy and salt were mixed well, and immobilized Bacillus subtilis was added to anchovy sauce prepared by aging and fermenting at room temperature for 10 months. The circulation pump was adjusted to circulate in the reactor for 5 hours and allowed to react for 2 weeks.

Comparative Example 1 Example 1 Histamine 2700 ppm 1503 ppm

As a result of the experiment, it can be seen that the histamine content of Example 1 in which the immobilized Bacillus subtilis was put into the fermented product during the manufacturing process, as shown in Table 2, was lowered.

Therefore, the immobilized Bacillus subtilis has an effect of removing the amine component contained in the fermented seafood, and thus a fermented product having a low histamine content can be produced.

Hereinafter, experimental results for examining the effect of removing the histamine component of the immobilized Bacillus subtilis in the method of reducing the biogenic amine in the present invention will be described in detail.

히 Histamine decomposition effect of immobilized Bacillus subtilis

In order to examine the effect of removing the histamine component of the immobilized Bacillus subtilis in the method of reducing the biogenic amine according to the present invention, the salt solution containing the histamine solution was filled in the reactor and the histamine content was measured with the immobilized Bacillus subtilis.

The concentration of histamine in the salt solution mixed with the histamine solution used in this experiment was 100 ppm, and the amount of histamine was measured with the time of immobilized Bacillus subtilis.

Reaction time (days)
Histamine ppm 0 100 10 85.6 20 72.7 30 58.5 50 63.6 60 53.2

As shown in Table 3, it can be seen that histamine content decreases with an increase in reaction time.

Therefore, it can be seen that the histamine component contained in the high salinity fish sauce or fermented seafood is removed by immobilized Bacillus subtilis, and the histamine content is gentle after 30 days.

In the following, the experiment contents for examining the change in the number of immobilized Bacillus subtilis grown at different salinity in the method of reducing the biogenic amine in the present invention will be described in detail.

히 Histamine decomposition effect of immobilized Bacillus subtilis according to salinity concentration

In the following experiment, in order to investigate the change of immobilized Bacillus subtilis according to the salt concentration in the method of reducing the biogenic amine according to the present invention, immobilized Bacillus subtilis was filled into the reactor at a volume ratio of 20% Lt; / RTI &gt; The reaction time was set to 24 hours in the reactor and the reaction was carried out for several days.

[Comparative Example 2]

In Comparative Example 2, immobilized Bacillus subtilis was grown under the condition that the salt concentration was 0%.

[Example 2]

In Example 2, immobilized Bacillus subtilis was grown under the condition that the salinity concentration was 7.5%.

[Example 3]

In Example 3, immobilized Bacillus subtilis was grown under a condition of a salt concentration of 15%.

[Example 4]

In Example 4, immobilized Bacillus subtilis was grown under the condition that the salt concentration was 20%.

Salinity% Number of immobilized Bacillus subtilis (days) 5 days 10 days 20 days 30 days 60 days 0 1.5 x 10 ² 4.8 x 10 ³ 4.6 × 10 ⁵ 5.2 × 10 ⁵ 5.3 × 10 ⁵ 7.5 1.7 x 10 ² 3.4 × 10 ³ 3.8 × 10 ³ 4.9 × 10 ⁶ 4.6 × 10 ⁵ 15 1.8 x 10 ² 3.5 × 10 ³ 2.2 x 10 ⁴ 4.4 × 10 ⁵ 4.5 x 10 ⁴ 20 1.5 x 10 ² 2.2 x 10 ² 3.6 × 10 ⁴ 3.5 × 10 ⁵ 4.2 × 10 ⁴

As shown in Table 4, it can be seen that the higher the salinity, the smaller the number of immobilized Bacillus subtilis that grows.

In the initial reaction, immobilized Bacillus subtilis grew slowly due to the inhibition of salinity. However, when the reaction start date reached 30 days, the maximum growth was observed in all the reactors even at high salt concentration.

As described above, it is to be understood that the technical structure of the present invention can be embodied in other specific forms without departing from the spirit and essential characteristics of the present invention.

Therefore, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than the foregoing description, All changes or modifications that come within the scope of the equivalent concept are to be construed as being included within the scope of the present invention.

S1. Bacillus subtilis culture
S2. Preparation of sodium alginate mixed solution
S3. Bead manufacturing
S4. Fixed Bacillus Carrier Molding
S5. Immobilization of Bacillus subtilis
S6. Adaptation phase
S7. Step of decomposition of biogenic amine

Claims (4)

Bacillus subtilis is cultured in a culture medium having a salt concentration of 1 to 5%
Shaking culture at a temperature of 30 to 35 占 폚 and 200 rpm for 12 hours;
A second step of preparing a mixed solution of sodium alginate by adding the above-mentioned cultured Bacillus subtilis to 1 to 4% sodium alginate solution;
A third step of dropping the sodium alginate mixed solution into a calcium chloride solution having a concentration of 0.5 to 3 moles to prepare beads;
A fourth step of curing the beads in a calcium salt solution for 30 minutes to form an immobilized bacterium carrier;
A fifth step of introducing a salt solution having a salt concentration of 5 to 20% into the reactor filled with the immobilized Bacillus subtilis carrier of the fourth step and activating the immobilized Bacillus subtilis through a salt solution adaptation period for 8 to 12 days;
A sixth step of adapting the immobilized Bacillus subtilis activated through the salt solution adaptation period of the fifth step to the fish sauce for 1 to 2 weeks;
The immobilized Bacillus subtilis adapted to the fish sauce of the sixth step is supplied to the fish sauce to decompose the amines,
And a seventh step of circulating the immobilized Bacillus subtilis adapted to the fish sauce in the reactor every 12 to 24 hours using a peristaltic pump to react with the biogenic amine component in the fish sauce for 1 to 3 months. Amine reduction method The method according to claim 1,
Wherein the culture liquid comprises yeast concentrate, mannitol, salt, ammonium phosphate, magnesium sulfate, distilled water, and a ratio of 1 to 10% based on the total weight of the raw materials for manufacturing the fermented fish product. The method according to claim 1,
Wherein the volume ratio of the immobilized Bacillus subtilis carrier packed in the reactor in the fifth step is 10 to 20%
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