JPH0956360A - Production of soy sauce - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a soy sauce prevented from growth of film yeast and halophilic Lactobacillus lactis, growth of which becomes a problem particularly in soy sauce of low salt content, also prevented from deterioration, and has high storage stability. SOLUTION: The objective soy sauce is obtained by adding to soy sauce a Monascus-cultured product or its processed product under such condition that, when in the figure the ordinate is assumed as anti-microbial activity (U/ml) of the Monascuscultured product or its processed product, and the abscissa as the salt concentration (w/v%) of soy sauce, as to fall within the shaded area above a slant line drawn by mutually connecting points of (0.9U/ml:3%), (0.9U/ml:5%), (0.7U/ml:7%), (0.5-U/ml:8.5%), (0.2U/ml:11%) and (0U/ml:13%).

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to soy sauce having a high storage stability, which can prevent the growth of film-forming yeast and salt-tolerant lactobacillus, which are problems in soy sauce, especially low-salt soy sauce, and prevent the deterioration of soy sauce by the microorganisms. Manufacturing method.

[0002]

2. Description of the Related Art Conventionally, there has been known a method of adding a culture of Monascus oryzae or a processed product thereof to a heat-processed food (for example, noodle soup) to prevent deterioration and decay (hereinafter referred to as deterioration) due to bacteria and mold. (Japanese Patent Laid-Open No. 64-5459).

[0003]

However, this method vaguely describes that when the culture of Aspergillus niger or its processed product is added to general heat-processed foods, it can prevent spoilage due to bacteria and mold. Is only
There is no description as to whether or not it has an effect of preventing spoilage due to the membrane-producing yeast or salt-tolerant lactobacilli mixed in soy sauce. On the other hand, soy sauce usually contains 16 to 20% of salt, and it is known that it has a strong bactericidal power because it contains other organic acids and nitrogen components, but in recent years it has been 80% of normal concentration of salt for health consciousness. The amount of thin salt soy sauce below or the amount of reduced salt soy sauce below 50% is increasing. However, such low-salt soy sauce has a reduced sterilizing power of the soy sauce itself, and is contaminated with salt-tolerant lactobacilli belonging to the genus Lactobacillus or the yeast produced during the production or after opening the packaging container, Turbidity and precipitation from these microorganisms occur in soy sauce,
Alternatively, when the soy sauce is spoiled by forming a film on the surface of the soy sauce, and when it is contaminated with a salt-tolerant lactobacillus strain that decarboxylates the amino acids (aspartic acid, glutamic acid) contained in the soy sauce, the acid decay occurs. , Soy sauce research institute, Vol. 16, N.
o. 5,200 to 205 (1990)) From the above, even if the film-forming yeast and salt-tolerant lactobacillus are mixed in the reduced-salt soy sauce, the growth of these microorganisms is completely prevented, and the deterioration of soy sauce is prevented. The appearance of soy sauce that can be made was strongly desired.

[0004]

The present inventors have finally completed the present invention as a result of various studies to solve such problems. That is, in the present invention, when the vertical axis represents the antibacterial activity (U / ml) of the Aspergillus niger culture or a treated product thereof and the horizontal axis represents the salt concentration of soy sauce (w / v%), (0.9 U /
ml: 3%), (0.9U / ml: 5%), (0.7U
/ Ml: 7%), (0.5U / ml: 8.5%),
(0.2 U / ml: 11%), (0 U / ml: 13%)
The method for producing soy sauce is characterized in that the culture of Aspergillus niger or the processed product thereof is added to the soy sauce under the condition of the range belonging to the upper diagonal line of the line drawn by connecting the points.

Hereinafter, the present invention will be described in detail. First, examples of the microorganisms for obtaining the red yeast mold culture include any microorganisms belonging to the genus Monascus and having an antibacterial substance-producing ability. Particularly, Monascus purpleus (Monascu)
s purpureus), the same rubé
r) and microorganisms belonging to the same pilosus having a high ability to produce antibacterial substances are preferable.

[0006] These microorganisms have been used since ancient times in the manufacture of red wine (Chien kanchu) in China, and in Taiwan, they have been typical microorganisms used in the manufacture of red wine, red wine and red milk tofu. It is widely recognized in Japan and overseas as a microorganism that can be used safely in food hygiene. Preferable specific examples of the microorganism include Monascus spicy
ATCC 16434, 16666, 1
6775, Monascus Purpleus ATCC1638
6, IFO 5965, Monascus Louver ATC
C15670, same 16246, same 16366, same 181
99, IFO 9203, Monascus Piro-Sus I
FO 4520, ATCC16368, IFO 452
1 and the like. In addition, ATCC is American Type Culture Collection (American)
Type Culture Collection),
IFO is an Institute for Fe (Institute for Fe)
mentation Osaka (Fermentation Research Institute).

[0007] To obtain a culture of Aspergillus niger or a treated product thereof using these microorganisms, first, the above-mentioned microorganisms are inoculated and cultured in a nutrient medium to obtain a culture.

As the nutrient medium, a medium suitable for growth of the microorganism is used, and any medium such as liquid medium, solid medium, synthetic medium and semi-synthetic medium is used.

The carbon source of the medium may be any assimilable carbon compound or its content, for example, wheat, barley, oats, rice, corn, corn, millet,
Soba, bran, glucose, sucrose, starch,
Maltose, dextrin, glycerin and the like are used, and the nitrogen source may be any assimilable nitrogen compound or a content thereof, for example, soybean, defatted soybean, gluten, bran, germ, peptone, meat extract, Casein, Co
And the like, ammonium sulfate, and the like. Further, suitable inorganic salts such as phosphoric acid, potassium, magnesium and calcium can be appropriately used, and various organic substances and inorganic substances necessary for the growth of the bacterium can be added to the medium as necessary. . These raw materials may be used in combination of two or more if necessary.

It is preferable to use wheat alone or a medium to which wheat is added as the culture medium, and to culture the above-mentioned microorganisms because the amount of the produced and accumulated antibacterial substance can be significantly increased.

The conditions such as temperature, time, and aeration for solid culture are as follows.
After heat sterilization with a bud or the like, for example, after treating at 120 ° C. for 30 minutes, the above microorganism is inoculated and cultured, and at 20 to 40 ° C. for 3 to 20.
Cultivation is carried out for a day to obtain a culture in which mycelia are sufficiently propagated.

In the case of liquid culture, an appropriate nutrient source is added and dissolved in water and sterilized by heating with an autoclave or the like, and then the above-mentioned microorganism is inoculated and cultivated at 20 to 40 ° C. for 3 to 20.
Cultivation is carried out for a day to obtain a liquid culture in which the cells are sufficiently propagated.

In the present invention, the above solid culture and liquid culture are collectively referred to as Monascus culture. The above Aspergillus oryzae culture is
Since it has sufficient antibacterial activity as it is, it can be added to soy sauce moromi or product soy sauce. An extract obtained by treating the Aspergillus niger culture with water again, ethyl alcohol
Or an extract obtained by treating with an organic solvent such as ethyl acetate, a concentrated solution of these, a decolorizing solution with activated carbon, a deodorizing solution, or a dried product (powder) of them (in the present invention, these are The processed product of red yeast rice culture) has a characteristic that it can prevent the growth of the film-forming yeast and the salt-tolerant lactobacillus and prevent the deterioration of soy sauce by adding a small amount or without impairing the flavor and color.

In order to carry out the present invention, the above-mentioned red yeast mold culture or a processed product thereof is added to soy sauce. The amount added is as shown in Figure 1.
As shown in Figure 5, the vertical axis represents the antibacterial activity (U / ml) of the Aspergillus niger culture or its treated product, and the horizontal axis represents the salt concentration (w / v).
%), (0.9 U / ml: 3%), (0.
9U / ml: 5%), (0.7U / ml: 7%),
(0.5 U / ml: 8.5%), (0.2 U / ml: 1
1%) and (0 U / ml: 13%). It is necessary to perform the processing under the condition of the range belonging to the upper hatched portion of the line drawn by connecting the points. This is extremely important in the present invention, and if the amount is less than this range, the purpose of the present invention is to prevent growth of the film-forming yeast and salt-tolerant lactobacillus that easily occur in soy sauce and prevent spoilage. Can not. The soy sauce referred to in the present invention refers to brewed soy sauce such as concentrated soy sauce, light mouth soy sauce, white soy sauce, reservoir soy sauce, regenerated (kanro) soy sauce, and raw soy sauce produced by a normal soy sauce brewing method, and a protein raw material is hydrolyzed with hydrochloric acid. Amino acid solution (chemical soy sauce) obtained by decomposing, neutralizing, filtering, and producing, semi-chemical semi-brewed soy sauce such as Shinshi No. 2 soy sauce, and soy sauce koji are put in low salt water and digested in a short time under heating. However, soy sauce-like seasoning obtained by adding yeast, lactic acid bacteria, etc. to it and fermenting it, protein raw materials (soybean, wheat gluten, corn gluten, isolated protein, etc.) alone or with a mixture of starchy raw materials It means a soy sauce-like seasoning (vegetable protein hydrolyzate) obtained by hydrolyzing with various enzymes such as proteolytic enzymes in the presence of salt or salt-free, and then performing fermentation with yeast, lactic acid bacteria or the like as it is.

The method for measuring the antibacterial activity of the present invention is shown below.

(1) Preparation of culture solution of membrane-producing yeast (a) Membrane-producing yeast: Tigosaccharomyces ruxii SK
-1 (FERM P-14990) (b) Liquid nutrient medium (composition) Glucose: 2.0% East extract: 1.0% Polypeptone: 2.0% pH: 5.0 (c) 2 Double concentration liquid nutrient medium Glucose: 4.0% East extract: 2.0% Polypeptone: 4.0% pH: 5.0 (d) Culture conditions: 30 ° C., 72 hours, shake culture ( 120
rpm) (e) Incubator: test tube having an inner diameter of about 16 mm and a length of 185 mm (f) Cultivation: 1 platinum loop of the membrane-producing yeast described above is placed in a liquid nutrient medium 5
The culture solution was obtained by inoculating and culturing in a ml.

(2) Preparation of a sample for measuring antibacterial activity (a) Using purified water obtained by subjecting distilled water to an ion exchange treatment, an antibacterial liquid (eg, an extract) derived from a culture of Aspergillus niger or its processed product A 2-fold dilution series was prepared and used as a sample for antibacterial activity measurement.

(3) Measurement of activity (a) Microplate (for example, a measuring portion measuring 8.5 cm in length and about 13 cm in width has a hole diameter of 6.4 mm and a depth of 1).
A transparent plastic plate in which 1.0 mm holes are regularly opened 96 times in total) is added to the above "double concentration liquid nutrient medium".
50 μl was dispensed, and 50 μl of each of the above “antimicrobial activity measurement samples” was added. (B) The "membranous yeast-producing medium" was diluted 100-fold with the "liquid nutrient medium", and 10 µl of each was inoculated into the microplate, followed by culturing at 30 ° C for 3 days, and then the medium. The presence or absence of sedimentation on the bottom and the formation of a film on the surface of the culture solution was observed with the naked eye. The activity (c) was determined from the following formula by determining the maximum dilution ratio (n) of the sample in which the growth of the membrane-producing yeast was inhibited. (D) Antibacterial activity (U / ml / antibacterial liquid) = n × 2

[0019]

[Experimental example]

(Example of production of reduced-salt soy sauce that can prevent spoilage due to film-forming yeast)
A soy sauce ripening moromi was obtained by a normal soy sauce brewing method, and this was squeezed and filtered to obtain a thick soy sauce. The soy sauce is desalted by an electrodialysis membrane, and then processed to produce a salt concentration of 3.0%.
0%, 7.0%, 8.5%, 11.0% and 13.0%
Thus, 6 kinds of low-salt soy sauce having 1.6% total nitrogen (w / v) and 3.5% ethyl alcohol (w / v) respectively were obtained. Each of these soy sauces is divided into 16 categories, and for each category,
A freeze-dried product of a water extract of the Aspergillus oryzae culture prepared by the following method was added so as to have a concentration of 0 to 1.5 U / ml as shown in FIG. 1 to obtain a test salt-reduced soy sauce. This soy sauce was inoculated with the film-forming yeast at a rate of 10 ⁵ cells / ml, and this was incubated at 30 ° C,
The soy sauce was stored for 15 days, and the soy sauce was observed with the naked eye to determine the antibacterial activity (U / ml) of the section in which the growth of the film-forming yeast was inhibited,
When the relevant portion is indicated by diagonal lines, FIG. 1 was obtained. FIG.
From the results, when adding freeze-dried water extract of red yeast rice culture to low salt soy sauce, the vertical axis is the antibacterial activity (U / ml) of red yeast rice culture or its processed product, and the horizontal axis is soy sauce. When the salt concentration (w / v%) of (0.9 U / ml: 3
%), (0.9 U / ml: 5%), (0.7 U / ml:
7%), (0.5 U / ml: 8.5%), (0.2 U /
(ml: 11%), (0U / ml: 13%), the storage stability that can prevent spoilage by membrane-forming yeast when performed under the conditions that belong to the upper hatched part of the line drawn by connecting the points. It can be seen that low salt soy sauce with a high content can be obtained.

(Preparation of lyophilized product of water extract of red yeast rice solid culture) To 50 g of crushed wheat, 35 ml of water was added and uniformly absorbed, and the mixture was put in a Ferbach flask having a capacity of 500 ml and placed in an autoclave. Heat sterilization (121 ℃, 30 minutes)
Processed. To this, pre-cultured Monascus monascus
A seed culture of species (sp.) ATCC 16775 was inoculated and cultured at 30 ° C. for 14 days to obtain a solid culture of red mold. The antibacterial activity of this culture was 15 U / g.culture. This culture was mixed with 4 times by weight of water, stirred every 1 hour, extracted for a total of 4 hours, and filtered through filter paper to obtain an extract. The antibacterial activity of this extract was 4.0 U / ml. This was freeze-dried by a conventional method to obtain 240 U / g-dried product, which was subjected to an addition test.

[0021]

Example 1 (Production Example of Soy Sauce with Excellent Deterioration Ability by Membrane-Producing Yeast) An antibacterial agent prepared by the above method was added to 100 liters of soy sauce aged moromi just before squeezing, which was obtained by a conventional method for producing thick soy sauce. 5.5 kg of Aspergillus oryzae culture having an activity of 15 U / g was mixed and squeezed and filtered by a conventional method to obtain an antibacterial activity of about 1 U / ml.
Moromi juice was prepared, desalted with an electrodialysis membrane, and produced to have a salt concentration of 8.5% and total nitrogen of 1.60.
%, Ethyl alcohol 3.5% reduced salt soy sauce was obtained. This reduced-salt soy sauce was inoculated with 10 ⁵ yeast cells / ml at 30 ° C. and stored for 15 days, and internal turbidity, bottom precipitation and film formation on the surface were completely observed. In other words, it was possible to obtain a soy sauce having high storage stability by inhibiting the growth of the film-forming yeast and preventing spoilage. For comparison, soy sauce was inoculated with the membrane-producing yeast in exactly the same manner except that the Aspergillus oryzae culture was not added, and the mixture was stored at 30 ° C. and the progress was observed. A film was observed, and it was confirmed that a film was formed on the entire surface on the 7th day.

From these results, it is known that the normal salt-reduced soy sauce has a reduced bactericidal activity of the soy sauce itself, and when it is contaminated by the film-forming yeast after opening, the microorganisms propagate and form a film on the surface of the soy sauce. Then, the soy sauce deteriorates, but if the Aspergillus niger culture or its processed product is added to the reduced salt soy sauce in advance, the growth of the soy sauce will be inhibited even if it is contaminated with the membrane-producing yeast, and the storage stability is high. It turns out that soy sauce is obtained.

[0023]

[Example 2] (Production example of soy sauce having high storage stability capable of inhibiting the growth of salt-tolerant lactobacillus) A salt-tolerant lactobacillus belonging to the genus Lactobacillus and decarboxylated from glutamic acid, which was isolated from acid-reduced soy sauce with reduced salt A strain having the property of being isolated was isolated. Then
This salt-tolerant lactobacillus was added to the reduced-salt soy sauce obtained in Example 1
0 ⁵ was inoculated so that / ml, which 30 ° C., 60
After storage for a day, no turbidity inside and no precipitation of cells near the bottom were observed, and growth of salt-tolerant lactobacilli was prevented, and a reduced salt soy sauce with high storage stability was obtained. From this result, it is understood that according to the present invention, even if salt-tolerant lactobacillus is mixed in, soy sauce having high storage stability can be obtained by inhibiting growth and preventing spoilage.

[0024]

EFFECTS OF THE INVENTION According to the present invention, soy sauce having extremely high storage stability can be obtained without adding a synthetic preservative, which can prevent the growth of the membrane-producing yeast and prevent spoilage. Especially 3%
It has a very excellent effect on soy sauce having a low salt concentration of (w / v) or more and less than 13%. Usually, in low salt soy sauce, ethyl alcohol is forced to be added so as to have a final concentration of 5 to 6% in order to prevent spoilage due to the film-forming yeast, but according to the present invention, this alcohol is used. It is possible to reduce the amount of alcohol to 3 to 4%, and it is possible to prevent flavor deterioration (alcohol odor, etc.) accompanying the addition of ethyl alcohol.

[Brief description of drawings]

FIG. 1 Antibacterial activity (U
/ Ml) and the salt concentration (w / v%) of soy sauce.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Tadanobu Nakadai, 339 Noda, Noda, Chiba Prefecture, Kikkoman Corp. (72) Inventor, Katsumi Yuasa, 339, Noda, Noda, Chiba, Kikkoman Corp.

Claims (2)

[Claims] 1. The vertical axis represents the antibacterial activity (U / ml) of the Aspergillus niger culture or a treated product thereof, and the horizontal axis represents the salt concentration of soy sauce (w / ml).
v%), (0.9 U / ml: 3%),
(0.9 U / ml: 5%), (0.7 U / ml: 7
%), (0.5 U / ml: 8.5%), (0.2 U / m
(11: 11%), (0U / ml: 13%), and the soy sauce is added with the Aspergillus niger culture or a treated product thereof under the condition of belonging to the upper hatched part of the line drawn by connecting the points. A method for producing soy sauce, which comprises: 2. The method for producing soy sauce according to claim 1, wherein low salt soy sauce having a salt concentration of 3% or more and less than 13% is used.