JP3479716B2 - Producing protein seasoning liquid - Google Patents

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to an improved method for producing a protein seasoning liquid, more specifically, a salt-free amino acid seasoning having a high amination ratio and a high utilization ratio by subjecting an animal and plant protein raw material to an alkali immersion treatment in advance. The present invention relates to a technique for producing a liquid.

[0002]

2. Description of the Related Art Conventionally, seasoning liquids containing soy sauce, miso and other proteolytic products as taste components have been known. However, these require the use of a considerably large amount of salt. Causes various restrictions. That is, soy sauce is generally inoculated with seed koji to koji raw material mainly consisting of steamed soybeans and roasted wheat flour to make koji, and then mixed with salt water to prepare koji, and ferment the resulting moromi to squeeze juice. It is produced as raw fried soy sauce, but when the koji is charged, it is usually about 23% so as to prevent spoilage due to the propagation of microorganisms derived from koji (miscellaneous bacteria) and to allow good fermentation and aging of moromi.
High and low salt concentrations are used before and after, which on the one hand gives a taste peculiar to soy sauce, but on the other hand, the salt concentration of the product is as high as 16 to 17%, which causes restrictions on its use.

Further, conventionally, a method for producing an animal or plant extract by allowing a protein-solubilizing enzyme to act on an animal or vegetable protein raw material has been known. However, when such a protein-solubilizing enzyme is used, the obtained extract is bitter. It has a fatal drawback,
It is difficult to use it as a seasoning liquid. The bitterness can be eliminated by using exo-type protease, but the exo-type protease itself is very difficult to separate and purify, and it has not yet been commercialized. There was no case where it was used for the production of a seasoning liquid from.

On the other hand, animal and plant protein hydrolysates (HAP,
HVP), that is, a seasoning obtained by decomposing animal and plant proteins with hydrochloric acid has been conventionally known, and the demand thereof is gradually increasing as a part of natural seasonings (auxiliary seasoning) used for instant foods and the like. However, such HAP, H
Regarding VP, recently, organochlorine by-products such as monochloropropanediol and dichloropropanediol, which are produced when the above proteins are hydrolyzed with hydrochloric acid, are mutagens, that is, genes are damaged and cells are mutated. It has been confirmed that the substance becomes a substance that causes aging, and there is a concern that toxic formic acid and the like may be mixed in, and its safety is becoming a problem.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a new salt-free protein seasoning solution which is safe and has no salt and has an excellent bitter taste, instead of the above acid-decomposable HVP and HAP. It is in.

As a result of earnest studies, the present inventor first found that an enzyme preparation (exo-type peptidase) obtained by self-digesting a protein-solubilizing enzyme and an alcohol-soaked product of microbial solid culture koji to an edible animal protein raw material. It was found that a salt-free and safe seasoning liquid which meets the above-mentioned purpose and is safe, excellent in taste, and free from bitterness and having good quality can be obtained by the action of the above. -1
25734, Japanese Patent No. 2932130).

As a result of further researches conducted by the present inventors, in the above-mentioned method, when the animal or plant raw material was previously subjected to the alkali immersion treatment, amino acids, especially glutamic acid, which has the most important influence on the improvement of the taste, While discovering the fact that the production yield of aspartic acid is dramatically improved, such an alkaline immersion treatment is carried out by an enzymatic hydrolysis treatment such as using a protein solubilizing enzyme and a microbial solid culture koji as a protein raw material. It was confirmed that the protein seasoning solution was effective throughout the production, and the present invention was completed here.

[0008]

According to the present invention, there is provided a method for producing a protein seasoning liquid, which comprises subjecting an animal or plant protein raw material to an alkali immersion treatment and then an enzymatic hydrolysis treatment.

Further, according to the present invention, the animal and plant protein raw material is preliminarily immersed in an alkali, the protein solubilizing enzyme is allowed to act under the temperature condition of 15 ° C. or lower and in the presence of alcohol to perform primary decomposition, and then the microbial solid culture koji is used. The method for producing a protein seasoning solution is characterized in that the protein is subjected to secondary decomposition under the same conditions.

Particularly preferred embodiments of the method of the present invention include those in which the alkali dipping treatment is carried out for at least 30 minutes under conditions of pH 10 to 12 and those in which the alkali dipping treatment is carried out in the presence of alcohol. .

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION According to the method of the present invention, the solubilization rate and the amination rate of a raw material protein are remarkably improved, and the amount of amino acids produced, particularly the content of glutamic acid and aspartic acid as taste components is greatly improved. A high seasoning liquid can be obtained. The reason for this is not completely understood at present, but the alkaline immersion treatment causes the tissue of the raw material protein to become soft and susceptible to enzymatic action, and the inhibitor of the biological enzyme contained in the raw material protein itself is inactivated. It is thought that this is due to the fact that it becomes susceptible to enzymatic action. Further, according to the above alkali immersion treatment, it is possible to prevent spoilage, sterilization and freshness of the raw material protein, and there is an advantage that the treatment such as frozen storage is unnecessary.

The seasoning liquid obtained by the method of the present invention, as it is or after being concentrated or pulverized as necessary, is used for a wide variety of processed foods or food materials such as instant foods, paste products, hams and sausages, pickles. It can be widely applied to various uses such as, dressing, sauce, beverage, etc. By the application, the taste of processed food or food material can be improved, and supplement of protein nutrition can be performed.
In particular, according to the method of the present invention using a specific enzyme agent derived from a microbial solid culture koji such as soy sauce koji, there is a rich body in which the content of glutamic acid and the like, which is the main ingredient of umami, is safe and salt-free. A protein seasoning liquid having an excellent taste can be obtained.

The method of the present invention will be described in detail below. The method of the present invention is characterized by preliminarily immersing an animal or plant protein raw material in an alkali for the production of a protein seasoning solution.
Here, as the animal and vegetable protein raw material, various things containing protein and provided for food, for example, vegetable edible raw materials such as grains and corn meal typified by wheat gluten, seafood, meat, etc. Examples thereof include processing by-products such as seafood, cut meat produced during the processing of meat, and animal food materials such as broth during the production of dried bonito and dried sardines. They are used in the form of an aqueous liquid suitable for the action of the enzyme, that is, an aqueous solution, an aqueous dispersion, a suspension or the like. In particular, in the present invention, it is possible to use, as the animal or plant raw material, chicken moth, bonito flakes, fish head coarse, etc., which have been conventionally discarded or used only as a feed for livestock, and are effective as food resources. It is available. It is preferable that the chicken roe etc. be finely pulverized in advance.

The alkali dipping treatment can be carried out by immersing and immersing the protein raw material as it is or by previously pulverizing it in an alkali aqueous solution such as sodium hydroxide aqueous solution.
At this time, the pH of the aqueous solution is preferably adjusted so that the pH after the protein raw material is added and immersed is about 10-12. The immersion time can be appropriately determined depending on the type of protein raw material and is not particularly limited, but it is preferably selected from the range of about 30 minutes or more and up to about 7 days. About time, preferably about 2
It is recommended to be set to about 4 hours. When bones, skins, and ligaments are used as the protein raw material, it is preferable that the time is about 1 to 7 days. The alkali immersion treatment can be usually performed at room temperature.

In the above alkali dipping treatment, alcohol such as ethanol can be added and blended in the alkaline aqueous solution, which can surely prevent the decomposition of the protein raw material. The amount of the alcohol added and compounded can be generally about 8 to 10%.

In a preferred embodiment of the method for producing a seasoning solution of the present invention, the animal-vegetable protein raw material that has been subjected to the alkali immersion treatment is treated with a protein-solubilizing enzyme for primary decomposition, and then a specific enzyme agent is caused to act on it. Next, a method of decomposing can be mentioned.

As the protein-solubilizing enzyme, any of various known ones commonly used in this field can be used. Typical examples thereof are endo-type proteases, including proteases derived from various microorganisms such as bacteria, molds, yeasts, actinomycetes, rennin, proteases derived from animals such as pancreatin, plant origins such as papain, bromelain and huaisin. Proteases and the like. Of course, one kind thereof may be used alone, or two or more kinds may be used in combination. As the protein solubilizing enzyme,
Since the pH of the raw material liquid (alkaline dipping treatment liquid) is alkaline, alkaline protease is preferable, but neutral protease or the like can also be used. The amount of the above enzyme used can be appropriately determined depending on the type of the enzyme used and the like, and is not particularly limited, but usually about 5 per g of protein.
00 to 10000 units, preferably 2000 to 8000
It is good to select from the range of about a unit. In the case of an endo-type neutral protease, the above activity unit uses milk casein as a substrate at pH 7.0 at 30 ° C. for 1 minute per minute.
It is calculated assuming that the titer for producing a decomposition product of μg of tyrosine is 1 unit.

The primary decomposition with the solubilizing enzyme agent is generally carried out by applying the above-mentioned suitable amount of enzyme to the raw material at 15 ° C. or lower, usually about 4 to 15 ° C., preferably about 8 to 12 ° C. It is carried out by reacting for about 1 hour to 7 days, and thus, an oligopeptide having an amino acid average chain length of about 5 to 15 (soluble at a final trichloroacetic acid concentration of 0.17 mol) is converted into an oligopeptide containing as much protein as possible. It can be hydrolyzed.

The primary decomposition can be carried out more preferably in the coexistence of about 8 to 10% of alcohol such as ethanol.

In the method of the present invention, after the completion of the above primary decomposition, the microbial solid culture koji is used as a secondary decomposition enzyme agent for secondary decomposition.

As typical examples of the enzyme agent (koji in which microorganisms are solid-cultured) used here, various well-known soy sauce koji and miso koji such as dark mouth soy sauce koji, light mouth soy sauce koji and accumulated soy sauce are used. Koji etc. can be illustrated. These are obtained by inoculating and culturing a koji material mainly consisting of steamed soybeans as a koji mold with koji molds, that is, Aspergillus oryzae, Aspergillus sojae and other bacteria belonging to the genus Aspergillus. In the present invention, the present invention is not limited to these, and various other well-known microorganisms that have been conventionally used for food, such as Aspergillus niger and Penicillium genus (Penicilium).
cillium, typically Penicilliu that attaches to beetles
Similarly, solid cultures of Rhizopus (Rhizopus, Rizopus niveus used for tempeh, etc.), Bacillus (Bacillus subtilis used for paste products, etc.), that is, their koji are also advantageous. Available for The above Bacillus solid culture also includes natto. The solid medium for culturing various microorganisms such as soybean may be soybean similar to soy sauce koji, and various nutrients that do not hinder the growth of each microorganism, such as various proteins, sugars, and fibers. It is also possible to appropriately select and contain a class or the like to have characteristics derived from the raw material.

PH of the liquid on which the enzyme agent (koji) acts
Will vary depending on the type of raw material and the conditions under which the protein solubilizing enzyme was acted, but in general, due to the action of the protein solubilizing enzyme described above, it decreased to around neutral, which is suitable for expressing the activity of this enzyme preparation. Therefore, there is no need for any special adjustment. Of course, if desired, a pH suitable for the action of the enzyme used in this step
It can also be adjusted to the conditions. The amount of the enzyme preparation used is
Although it can be appropriately determined, it is usually selected from the range of about 0.1 to 2 g per 1 g of animal and plant protein raw material.
In addition, the condition for causing the enzyme agent to act is generally 15 ° C.
Below, preferably at a temperature of about 10 to 14 ° C., about 30
It is preferably about 75 days. Thus, it is possible to produce a seasoning liquid having an excellent taste according to the present invention.

In the method of the present invention, the protein-solubilizing enzyme is first allowed to act on the raw material liquid that has been subjected to the alkali immersion treatment as described above,
Next, a specific enzyme agent is allowed to act, but in some cases, both enzymes are charged at the same time as the alkali-immersed raw materials, and both enzymes act on the raw materials in one step to obtain the desired seasoning liquid. It is also possible, and such methods are also included in the present invention.

The seasoning liquid of the present invention thus obtained can be used as it is for various purposes, and if necessary, after-treatments are carried out according to various known methods to obtain products suitable for various purposes. You can also

As the above-mentioned post-processing means, for example, the following means and combinations thereof can be appropriately adopted. (1) By solid-liquid separation treatment such as centrifugation and filtration,
Remove solids, etc. (2) The enzyme used is deactivated by heat treatment (usually at 70 to 100 ° C. for about 5 to 30 minutes). (3) Purification such as activated carbon treatment is performed to remove unnecessary odors. (4) Concentrate so that the solid content concentration is about 30 to 70%. (5) A powdery seasoning is prepared by a drying means such as spray drying.

[0026]

EFFECTS OF THE INVENTION According to the method of the present invention, spoilage and the like can be prevented in the absence of salt, and the solubilization rate and amination rate of a protein raw material can be dramatically improved, resulting in no salt, safe and umami. It is possible to produce a seasoning liquid of excellent quality with a markedly improved richness.

[0027]

EXAMPLES Hereinafter, test examples and examples will be given in order to explain the present invention in more detail.

The component analysis and the like of the sample and seasoning liquid in each example were determined by the following methods. (1) TN (total nitrogen content, g / dl): by Kjeldahl method. (2) FN (formol nitrogen, g / dl): According to the formol titration method. (3) Salt content: According to the Mohr method. (4) Dissolution rate (%): It is indicated by the ratio (percentage) of TN of the filtrate obtained by filtering the reaction solution with filter paper, relative to TN of the reaction solution. (5) Decomposition rate (%): Displayed as the ratio (percentage) of FN of the filtrate obtained by filtering the reaction solution with filter paper, relative to FN of the reaction solution. (6) Utilization rate (%): Displayed as a ratio (percentage) of TN × filtrate amount of the above filtrate to TN of all raw materials. (7) Amino acid analysis: With an amino acid analyzer (manufactured by Hitachi, Ltd., model 835 high-speed amino acid analyzer).

[0029]

[Test Example 1] pH and time of alkali dipping treatment Dry powder of dried bonito flakes was used as a protein raw material, and each 2
Caustic soda, water and alcohol (final concentration 10
%) To adjust each pH (total amount 1000 ml), divide each into 5 equal parts (200 ml each) and leave for 0, 1, 3, 5 and 7 days (immersion at 12 ± 2 ° C.), then add 2 each. Divide into equal parts and use alkaline protease (protein solubilizing enzyme agent, Orientase 22B, Hankyu Bio Industry Co., Ltd.)
(Manufactured by Mitsui Chemicals Co., Ltd.) and 8,000 units (per sample g) were added, and primary decomposition treatment was carried out at 12 ° C. for 5 days.

The TN and the dissolution rate of the filtrate obtained by filtering the obtained primary decomposition liquid were determined. The results are shown in Table 1 below.
Shown in.

[0031]

[Table 1]

From Table 1, according to the alkali dipping treatment according to the present invention, it is proportional to the dipping time and to the high pH during dipping, as compared with the case where it is not performed (dipping time: 0). Then, it can be seen that the TN and the dissolution rate are improved.

It has been found that particularly preferred immersion conditions in this example are pH 10-12 and 5-7 days.

[0034]

[Test Example 2] Enzyme reaction time and temperature during primary decomposition In Test Example 1, dried powder of dried bonito flakes (protein raw material)
The alkaline immersion treatment conditions were pH 11, temperature 12 ° C. for 5 days, and subsequent primary decomposition was performed using 8,000 units / sample g of alkaline protease (protein solubilizing enzyme agent, Orientase 22B, manufactured by Hankyu Bio Industry Co., Ltd.). Then, it was carried out at 4 to 24 ° C. for 1 to 7 days to examine an appropriate temperature and time for this primary decomposition treatment.

Table 2 below shows the results of the pH, TN and dissolution rate of the primary decomposition filtrate obtained under the prescribed temperature and time conditions.
Shown in.

[0036]

[Table 2]

From Table 2, it is preferable that the primary decomposition treatment following the alkali immersion treatment according to the present invention is carried out at 4 to 12 ° C., and the TN and the dissolution rate are improved as the treatment time is extended under this temperature condition. I knew that.

Further, when the temperature is raised, the pH of the primary decomposition solution is lowered in proportion to this (which is considered to be due to the production or decay of lactic acid by lactic acid bacteria), which causes the dissolution rate and TN to gradually decrease. I knew that I would do it.

[0039]

[Test Example 3] Type of raw material and soaking time The raw material for protein may be hard such as bone, muscle, toughness, skin, soft meat such as meat, bonito soup, soup, etc. A variety of substances are included, including water-soluble ones, and the degree of susceptibility to degradation by proteolytic enzymes varies. This test was conducted to investigate the relationship between the alkali immersion treatment time, the amount of enzyme agent used, and the solubility.

As protein raw materials, (1) bonito broth, (2) powdered gelatin, (3) wheat gluten, (4) fish meat portion, (5) beef meat portion, (6) okara and (7) shown in Table 3 below are used. Each of chicken cod (used as a reaction solution at each concentration indicated by TN in the table, 1
50 ml of each was subjected to the test).
Alkaline immersion treatment was carried out for a predetermined time (1 hour to 7 days) in the same manner as in Test Example 1, and then 12 ° C., alcohol 10%.
Primary treatment with alkaline protease was performed under the conditions of (final concentration) and 5 days (the enzyme is the same as in Test Example 1, and the amount added is shown in Table 3).

The meat portion as the raw material protein is minced beforehand, and the chicken cod is ground to a particle size of about 0.5 mm or less,
It was submitted to the test.

The results are shown in Tables 3 and 4.

[0043]

[Table 3]

[0044]

[Table 4]

From Tables 3 and 4, according to the alkali dipping treatment of the present invention, the TN and the dissolution rate by primary decomposition are remarkably improved in any case of various protein raw materials by appropriately adjusting the dipping treatment time. It turns out that it can be done.

[0046]

Example 1 The same various protein raw materials as those used in Test Example 3 (indicated by respective numbers (1) to (7) in Table 5 below) and further corn gluten powder (“(8)” in Table 5 below) ] (Each total 20 liters) was added to caustic soda, water and alcohol (final concentration 10%) to adjust the pH to 11, and each was immersed for a predetermined time (immersion temperature: 12 ° C.).

Then, a predetermined amount (unit / sample g) of the same alkaline protease as used in Test Example 1 was added and mixed, and a primary decomposition reaction was carried out for a predetermined time (reaction temperature: 1
2 ° C).

The primary decomposition solution thus obtained is divided into two equal parts,
On the other hand, the soy sauce koji (called "enzyme preparation A")
On the other hand, soya-based soy sauce koji (referred to as "enzyme preparation B") was added and mixed at a ratio of 1/3 times the weight of the protein raw material (converted to the solid content of the protein), and the mixture was kept at a temperature of 12 ° C for 2 months for 2 months. Subsequent decomposition reaction is performed to obtain the desired protein seasoning liquid (stock solution)
Got

The neutral protease activity of each of the above-mentioned oryzed soy sauce koji and soya type soy sauce noodles is about 800 units / g and about 880 units / g, and the acidic protease activity (similar to the neutral protease activity at pH 3.0). 280 units / g and about 310 units / g, glutaminase activity (37 L-glutamine as a substrate).
The unit of the number of mg of glutamic acid produced at 60 ° C. for 60 minutes is about 118 units / g and about 140 units / g.

The resulting seasoning solution stock solution is added with about 10 liters of activated carbon 0.3 to 0.5 kg and perlite 0.8 to 1.0 kg as a filter aid, and filtered using a small filter press. Therefore, the filtrate 7.8-8.
9 liters are obtained. The results of TN, FN, decomposition rate, pH and utilization rate (filtrate TN / reaction solution TN × 100) of this product are shown in Table 5 below.

[0051]

[Table 5]

From Table 5, the obtained seasoning liquid is TN, F
It can be seen that it is excellent in N, decomposition rate, pH and utilization rate.

In addition, the salt content of each seasoning solution was 0.
It was less than 06%.

Further, each of the obtained seasoning liquids was subjected to amino acid analysis using an amino acid analyzer (manufactured by Hitachi Ltd., model 835 high-speed amino acid analyzer), and the result was TN.
Table 6 below shows the amount of amino acid per 1.0 (mg / dl).
(Each seasoning liquid is indicated by the number of the protein raw material used).

Table 6 shows the results of the seasoning liquid (indicated by "(9)" in the table) obtained in the same manner as in Example 1 using the dried bonito extract meal powder used in Test Example 1. Write together.

In Table 6, as a control, commercially available soy sauce (denoted by "a" in the table), commercially available acid-decomposing amino acid-based seasoning solution (denoted by "b" in the table) and unsalted soy sauce (soy sauce koji with ethanol moromi) Aged at 12 ° C. for 2 months as described in JP-A-6-12
5735 gazette) (indicated as “c” in the table), the same amino acid analysis results are also shown.

[0057]

[Table 6]

From Table 6, all the seasoning solutions obtained by the method of the present invention have a high amino acid content, and in particular, the amounts of aspartic acid (Asp) and glutamic acid (Glu), which are the main components of the taste components, are remarkably high, which is excellent. It was clarified that it has a taste.

─────────────────────────────────────────────────── ─── Continuation of the front page (56) Reference JP-A-7-284369 (JP, A) JP-A-2001-61445 (JP, A) JP-A-2000-53696 (JP, A) (58) Fields investigated ( Int.Cl. ⁷ , DB name) A23L 1/20-1/238 A23J 3/34 A23L 1/305

Claims (3)

(57) [Claims] 1. An animal or vegetable protein raw material having a pH of 10-12.
A method for producing a protein seasoning solution, which comprises subjecting to alkaline soaking for at least 30 minutes under the following conditions and then subjecting to enzymatic hydrolysis. 2. The pH of the animal and vegetable protein raw material is adjusted to 10 to 12.
After at least 30 minutes alkali immersion treatment in that condition, 15
Production of a protein seasoning solution characterized in that a protein-solubilizing enzyme is allowed to act on it under the temperature condition of ℃ ℃ or less and in the presence of alcohol to cause primary decomposition, and then microbial solid culture koji is allowed to act under the same condition for secondary decomposition. Method. 3. A raw material for animal and vegetable proteins is a raw material for animal food.
(However, chicken egg protein is excluded) , The method of Claim 1 or 2.