JPH01108955A - Production of seasoning solution with high glutamic acid content - Google Patents

Abstract

PURPOSE:To obtain the above seasoning solution having thick tastiness, by inoculating a soy sauce KOJI (yeast) mold to a raw material for producing soy sauce, preparing a soy sauce KOJI, adding a halotolerant glutaminase- containing enzymic agent together with a saline solution to the resultant KOJI or adding the saline solution and then the enzymic agent thereto. CONSTITUTION:Defatted soybeans, wheat, etc., are treated by a conventional method to provide a raw material for producing soy sauce and a soy sauce KOJI (yeast) mold is inoculated thereinto to afford a soy sauce KOJI. An enzymic agent containing a halotolerant glutaminase, such as lipase, pectinase or protease, together with a saline solution, is added to the soy sauce KOJI or the saline solution is added and the enzymic agent is then added to provide the aimed enzymic agent.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a method for producing a seasoning liquid with a high glutamic acid content.

Generally, it is believed that the higher the glutamic acid content of soy sauce, the better the taste.

According to the method of the present invention, the content of glutamic acid in a seasoning liquid such as soy sauce is significantly increased, and a seasoning liquid with good taste can be obtained, which is of great benefit to the food industry.

(Prior art) It has been known to enzymatically hydrolyze raw materials for soy sauce production and bring the decomposition solution into contact with immobilized peptidase and/or immobilized glutaminase to increase the content of total amino acids and glutamic acid. .

(JP 61-85165) Furthermore, in the production of soy sauce, it is known that soy sauce koji is prepared in the coexistence of glutaminase-producing bacteria of the genus Rhodotorula to increase the glutamic acid content. (Tokuko Showa 5
7-55388) As described above, various attempts have been made to convert glutamine produced in soy sauce mash into glutamic acid using glutaminase to enhance the taste, but the purified product of glutaminase is expensive, and The drawback was that the production of glutaminase from viable cells was low.

(Problems to be Solved by the Invention) It is clear that if glutaminase is inexpensive and can reliably convert glutamine produced in soy sauce mash into glutamic acid, the taste of seasoning liquids such as soy sauce will improve.

(Means for Solving the Problem) The present inventors conducted extensive research in search of an inexpensive glutaminase that can be added to soy sauce mash to reliably convert it into glutamic acid, and found that conventionally commercially available lipases, proteases, etc. He learned that these enzymes contained a certain amount of glutaminase, and that if these enzymes were added directly to soy sauce mash, glutamine would be reliably converted to glutamic acid.

The present invention involves adding an enzyme agent containing a high titer of salt-tolerant glutaminase to soy sauce koji, which is produced by inoculating soy sauce koji mold into a raw material for soy sauce production that has been treated in a conventional manner and then making koji together with one common salt or after adding a saline solution. This is a method for producing a seasoning liquid with a high glutamic acid content.

The enzyme agent containing a high titer of salt-tolerant glutaminase used in the present invention may be any enzyme agent that has been confirmed to contain glutaminase, such as commercially available lipase, pectinase, and protease. Examples include enzyme agents.

0Aspergillus niger
"Lipase A 'Amano'" (manufactured by Ohno Pharmaceutical Co., Ltd.) derived from Aspergillus pulverulentus (Aspsrgillus pulveruJentu
s) derived from “Pectinase G “Amano”” (Ohno Pharmaceutical Co., Ltd.)
Proleather, a protease agent derived from Bacillus sp. (manufactured by Ohno Pharmaceutical Co., Ltd.)
Co., Ltd.) These enzyme preparations containing glutaminase are added to soy sauce koji together with or after adding the salt solution.

The amount of enzyme agent containing glutaminase to be added may be sufficient to convert glutamine produced by glutaminase into glutamic acid, but it is preferable to add an excess amount beyond this amount. Also as an enzyme agent.

Addition of about 0.5 to 1.5% of the weight of koji is sufficient.

Glutaminase contained in various enzyme preparations exhibits high activity over a wide pH range, with no decrease in activity even at pl (5.0 to 5.5). Since it maintains its activity even at a concentration of 20%, it can convert glutamine produced in soy sauce mash into glutamic acid, thereby improving the taste of seasonings such as soy sauce.

Next, test examples and examples of the present invention will be shown, and the general glutaminase measurement method used here is as follows.6 Glutaminase measurement method (GLDH-diaphorase method) Substrate (0,625% L-glutaminase method) , 0.05M KPB
p) 17.0) 2mm, enzyme sample solution 0.5+il
Add and mix. After reacting at 37°C for 60 minutes, 10
Inactivate by heating at 0°C for 5 minutes.

Take 0.2 mm of the reaction solution and measure the produced glutamic acid using an F-Kit L-Glutamine dish (manufactured by Boehringer Mannheim Yamanouchi Co., Ltd.).

For the enzyme blank, heat inactivate the same enzyme solution, perform the same operation as above, measure the absorbance of a492n+m,
The enzyme activity is calculated from the difference in absorbance.

The activity of producing 1 μ5 tole of monoglutamic acid per minute is defined as 1 unit.

Test Example 1 Effect of pH on glutaminase activity in enzyme preparation
% and dissolved, reacted at 37°C for 1 hour, and measured the relative activity of each glutaminase, obtaining the results shown in Figure 1.

2. Effect of temperature on glutaminase activity in enzyme preparations All pH 7 for each of the same enzyme preparations as in 1 above.
The glutaminase was dissolved in a 1% buffer solution and reacted at each temperature of 30 to 60°C for 1 hour, and the relative activity of each glutaminase was measured, and the results shown in Figure 2 were obtained.

3. Effect of salt on glutaminase activity in enzyme preparations Salt concentration 0 to 2 for each of the same enzyme preparations as in 1 above.
Add 1% to 0% buffer (pH 7,0) and dissolve.
The reaction was carried out at 37° C. for 1 hour, and the relative activity of each glutaminase was measured, and the results shown in FIG. 3 were obtained.

Note that, as a control, the relative activity of glutaminase was similarly measured for a mixture prepared by adding 5% soy sauce koji and stirring.

4. Thermostability of glutaminase activity in enzyme preparations in the presence of salt Salt concentration 10% for each of the same enzyme preparations as in 1 above
Add 1% to the buffer solution (pH 7,0) and dissolve.

After standing at 60° C. for 0 to 120 minutes, the relative activity of each glutaminase was measured, and the results shown in FIG. 4 were obtained.

As a control, a sample prepared by adding 5% soy sauce koji and stirring was subjected to the same treatment, and then the relative activity of glutaminase was measured.

Example 1 Soy sauce koji mold was inoculated into a mixture of defatted soybeans and wheat that had been steamed, roasted and crushed in a conventional manner at a ratio of 7:3, and 13% (w) /v) Salt solution 360-, enzyme agent Lipase A "Amano", Pectinase G "Amano" and Proleather are respectively 0.25%, 0.5%, 1.0% and 1.5% based on the weight of koji. % and was prepared and kept at 45°C for 7 days. Thereafter, a seasoning liquid was obtained by natural filtration using &2 filter paper.

For comparison, a seasoning liquid was prepared under similar conditions without adding enzyme.

The analysis of the seasoning liquid was conducted according to the "Soy Sauce Test Method" published by the Japan Soy Sauce Research Institute.

The analytical values were as shown in Table 1.

Example 2 7 kg of defatted soybeans and 3 kg of wheat were each steamed, roasted and crushed in a conventional manner and mixed, inoculated with soy sauce koji mold and made into koji to obtain soy sauce koji. 13% soy sauce koji cooled to 5℃ (
v/v) A saline solution was added to a stainless steel jacket tank to make 13 water, and 0.5% of Proleather was added to the soy sauce koji at the same time. The temperature was gradually increased to 30°C at a rate of 2°C/hour, and the temperature was maintained at 30°C for 3 days. Next, the temperature was gradually increased to 45°C at a rate of 2°C/hour, and the temperature was maintained at 45°C for 4 days. Salt-tolerant lactic acid bacteria were inoculated 24 hours after preparation, and stirring was performed to the extent that the strains were kept uniform. After completion of koji hydrolysis and lactic acid fermentation, the mixture was pressed using a small press to obtain a seasoning liquid.

As a comparative example, the soy sauce koji obtained above was hydrolyzed under the same conditions as above without adding an enzyme agent to obtain a seasoning liquid.

The results of measuring the increase in glutamic acid in this example as well as in the comparative example are shown in FIG.

Example 3 23% (w/v) saline was added to 3.2 kg of soy sauce koji obtained in Example 1 to make 12 water, and at the same time, the enzyme agents Lipase A "Amano" and Pectinase G "Amano" were added. and Proleather, each at 0.25% based on the weight of koji.
The fat taste was controlled for 4 months using a normal soy sauce brewing method with 0.5% and 1.0% added, and the various strains were pressed to obtain a seasoning liquid.

For comparison, raw soy sauce was prepared without adding any enzymes, and was managed and pressed using the same methods to obtain raw soy sauce.

The analysis results are shown in Table 2.

For the sensory test, 25 panelists conducted a taste test using the ranking method ("Soy Sauce Test Method" 1 Japan Soy Sauce Research Institute). As a result, it was revealed that each enzyme-added soy sauce was superior in terms of quality, with comparable taste and aroma, as well as rich umami, compared to commercially available soy sauces.

[Brief explanation of the drawing]

Figure 1 is a graph showing the effect of pH on glutaminase activity in an enzyme preparation in Test Example 1, and Figure 2 is a graph showing the effect of temperature on glutaminase activity in an enzyme preparation in Test Example 2. Figure 3 is a graph showing the influence of salt on glutaminase activity in an enzyme preparation, and Figure 4 is a graph showing the thermal stability of glutaminase activity in an enzyme preparation in the presence of salt in Test Example 4. FIG. 5 is a diagram in which the increase in glutamic acid in soy sauce mash was measured in Example 2 using the method of the present invention and a comparative example. Agent Patent Attorney Door 1) Parent Male Temperature ('C) NaC1 ('/,) S Question (#) 5th Zukei jL day! (Day)

Claims (1)

[Claims] It is characterized in that an enzyme agent containing a high titer of salt-tolerant glutaminase is added to the soy sauce koji produced by inoculating soy sauce koji mold to the raw material for soy sauce production processed in a conventional manner and making koji together with or after adding the salt water. A method for producing a seasoning liquid with high glutamic acid content.