JP2004201678A - Seasoning and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a seasoning obtained by decomposing defatted soybeans through a solid malt process and so improved that common salt is not contained or contained in a small amount, a high degree of conversion of amino acids is attained, and strong flavor components are scarcely contained. <P>SOLUTION: This seasoning is produced through a process (i) in which solid malted soybeans are prepared by inoculating a raw material containing the defatted soybeans with bacteria having ability of hydrolyzing proteins and another process (ii) in which a mash is formed by adding a brewing liquid to the prepared solid malted soybeans so that the common salt is contained in the mash in a specified concentration, and then the proteins are hydrolyzed by fermenting the formed mash. Further, lactic acid bacteria are added to the raw material in the process (i), and the lactic acid bacteria are, if necessary, added to the mash in the process (ii). Furthermore, the seasoning is produced so as to attain the degree of conversion of the amino acids of ≥65%, an isobutyl alcohol concentration of ≤0.1mg/nitrogen (g), and a normal butyl alcohol concentration of ≤0.25mg/nitrogen (g). <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

  The present invention relates to an amino acid composition, more specifically, an amino acid composition obtained by hydrolyzing a proteinous koji containing soybean protein as a main material and releasing 65% or more of the amino acids contained in the protein, and producing the composition. How to do it.

  BACKGROUND ART Conventionally, seasonings containing amino acids and peptides such as glutamic acid have been used to impart flavors such as umami and kokumi to various seasonings such as processed foods, soups and sauces, and pickles. Among these materials, those obtained by decomposing vegetable proteins such as soybeans and animal proteins such as beef, pork and chicken with acids or enzymes are often used. In particular, an amino acid solution (HVP: Hydrolyzed Vegetable Protein) obtained by acid-decomposing a vegetable protein and soy sauce which is a traditional fermented seasoning in Japan are representative.

  HVP is produced by hydrolyzing cereals rich in protein, such as soybeans, with hydrochloric acid at high temperatures, so that almost 100% of the protein is broken down into amino acids, and umami such as glutamic acid is produced. Contains a large amount. In addition, due to the decomposition reaction under acidic high temperature conditions, a substance that expresses the unique flavor and taste of HVP is produced from sugar, amino acids, organic acids, and lipids by a chemical reaction. For example, it is known that sotolone is produced as a flavor component, and formic acid and levulinic acid are produced as taste components.

  In a traditional method of producing soy sauce, first, defatted soybeans as a raw material are steamed, then mixed with an approximately equal amount of roasted wheat, and inoculated with seed koji to produce koji. The obtained koji is prepared in a salt solution, moromi, fermented and matured for a long time. The taste is due to amino acids and peptides, but since the decomposition of the protein has progressed only about 50%, the content of amino acids, particularly glutamic acid, is lower than that of HVP, and this affects the taste titer. In addition, soy sauce has a unique soy sauce flavor because it produces organic acids such as various alcohols, esters, and acetic acid by the action of yeast and lactic acid bacteria during aging for more than six months.

  As described above, HVP and soy sauce have been used in various processed foods and seasonings as an excellent basic seasoning material having a function of imparting a unique flavor, such as umami and kokumi, regardless of the east and west of the ocean, Since it is a liquid and contains 10 to 20% of salt, when used in a large amount, the salty taste becomes too strong, and there is a problem that umami and kokumi of the required strength cannot be exhibited. In addition, depending on the processed food used, since the salt contained affects the physical properties and taste of the processed food, the amount used is limited, and as a result, it is not possible to impart umami of the required strength and kokumi There are also problems. For example, fish paste such as kamaboko is known to solidify when a large amount of salt is added, that is, the so-called "sit" deteriorates, so that a large amount of HVP and soy sauce cannot be added. Therefore, it is desirable that the basic seasoning material that imparts umami and kokumi does not contain a large amount of salt.

  If we can make HVP and soy sauce that do not contain a large amount of salt and have umami and kokumi as they are, it will be possible to adjust the concentration of salt arbitrarily and it will be a food manufacturer more than ever In addition to increasing the chances of using umami, umami and kokumi can be widely added to processed foods and various seasonings. In addition, since unnecessary salt is not put into food as a result, there is an advantage that the salt intake of the consumer can be reduced. In order to reduce the intake of salt, there is a reduced salt soy sauce produced by desalinating the soy sauce of this brew, but there is a problem that the umami taste and richness of soy sauce also decrease with salt, It doesn't fit.

  In addition to the problem of salt in HVP and soy sauce, as mentioned above, HVP and soy sauce also have unique and strong flavors. There is a problem of breaking. In particular, since the flavors of other food materials and seasoning materials are masked, there is a problem that the complex flavor of the food demanded by the consumer is eliminated, and for example, only the soy sauce flavor becomes strong and faint. Mentsuyu is one such example. Mentsuyu is a mixture of 'dashi', which is mainly taken from dried bonito, and 'kaeshi', which is soy sauce. The flavor components of soy sauce, such as isobutyl alcohol (iba), normal butyl alcohol (nba), and isoamyl alcohol (iaa) There is a problem that the bonito dashi flavor is masked and the flavor becomes worse (Patent Document 1).

  Therefore, it is desirable that the basic seasoning material imparting a taste such as umami and kokumi not have a strong flavor. If such a seasoning with less flavor can be obtained, if HVP flavor or soy sauce flavor is required, HVP and soy sauce can be partially mixed and used. It is also possible to arbitrarily add an appropriate amount of flavor that does not mask the flavor of other food materials and seasoning materials.

  Until now, as flavorless soy sauce, deodorized soy sauce (Patent Document 1) in which the flavor components of soy sauce such as iba, nba, and iaa have been reduced by blowing nitrogen gas into soy sauce, and solid koji in the absence of salt. Soy sauce produced by decomposing at high temperature for a short time (Patent Document 2) has been developed.

  As described above, it is desirable that the basic seasoning material that imparts a taste such as umami or kokumi does not contain salt and does not contain a strong flavor component. It is desirable that the amino acids contained in the protein be released at least as much as soy sauce, and if possible, almost 100% as much as HVP. The high amino acid conversion rate of HVP is due to its production method, that is, acid decomposition. However, regulations on the 3-MCPD generated during the acid decomposition process have been tightened, for example, the upper limit has been set in Europe, and the production method of the basic seasoning material to be newly developed in the future is not acid decomposition, but the same traditional solid material as soy sauce. It is desirable to use the koji method.

  Summarizing the above situation, as a basic seasoning material that imparts umami and kokumi, it is manufactured by the traditional solid koji method, contains no salt, has an amino acid content per nitrogen as high as HVP, and has a strong flavor What does not contain components is desired.

  In the production of soy sauce by the solid koji method, a method of hydrolyzing koji at 2 to 25 ° C. without containing salt has been developed as a method for increasing the amino acid content (Patent Document 3). However, in this method, although the amino acid content is increased, there is a problem in that salt is added to the fermented koji after hydrolysis to ferment the resulting moromi, so that it contains salt. Also, a method of hydrolyzing a mixture of koji and yeast at 2 to 25 ° C. in the absence of salt is disclosed (Patent Document 4). However, since this method uses yeast, it naturally contains a flavor component.

  Further, a technique for producing soy sauce and a flavoring agent using Aspergillus oryzae having an endopeptidase and exonuclease activity twice or more higher than that of a wild strain of Aspergillus oryzae is disclosed (Patent Document 5). There is a problem of including.

By the way, when a fermented protein koji is prepared from a protein-containing material and a carbohydrate and the fermented protein koji is hydrolyzed at a temperature of 15 ° C. to 60 ° C. and a pH of 4.5 to 10 for 6 hours to 28 days, A method of inoculating 10 ³ to 10 ⁷ cfu of lactic acid bacteria per fermentation protein koji stage or hydrolysis stage is disclosed (Patent Document 6). Because hydrolysis at low temperatures in the absence of salt may risk the growth of undesirable microorganisms, this method involves ingesting a culture of lactic acid bacteria to protect the koji from the growth of said microorganisms. is there. However, since the growth of the inoculum in the inoculum during the koji production and the decomposition occurs, there is a problem that the amino acid released particularly during the decomposition is assimilated and the amino acid yield is reduced.

In addition, in the production of fermented seasonings, a method of making koji using a raw material obtained by lactic acid fermentation (Patent Document 7) is known. In this method, koji is charged using a saline solution.
Japanese Patent No. 2862719 Japanese Patent Application No. 2002-103013 U.S. Pat. No. 5,523,100 U.S. Pat. No. 5,888,561 US Patent No. 6,090,607 U.S. Pat. No. 5,965,178 Patent No. 3027352

  The present invention is a seasoning obtained by decomposing a vegetable protein by the same solid koji method as soy sauce, which does not contain salt, or has a low salt content, a high amino acid conversion rate, and almost has a strong flavor component. It is an object to provide a seasoning that does not contain, and a method for producing the same.

The present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, acetic acid in addition to aroma components such as isobutyl alcohol, normal butyl alcohol, and isoamyl alcohol contained in soy sauce, other seasonings and food materials It has been found that excellent flavoring can be obtained by masking the taste and flavor of, and reducing the content thereof. In the production of soy sauce, the presence of an appropriate amount of lactic acid bacteria in both the koji making and the hydrolysis (fermentation) steps of the koji makes it possible to obtain a seasoning in which the aroma component is reduced without adding salt. Successful.
That is, the present invention is as follows.

(1) A seasoning obtained by allowing a microorganism having the ability to hydrolyze a protein to act on a raw material containing a vegetable protein, wherein the amino acid conversion rate is 65% or more and the isobutyl alcohol concentration is 0.1 mg / g · nitrogen or less. A seasoning having a normal butyl alcohol concentration of 0.25 mg / g · nitrogen or less, an isoamyl alcohol concentration of 0.5 mg / g · nitrogen or less, and an acetic acid concentration of 100 mg / g · nitrogen or less.
(2) The seasoning according to (1), wherein the raw material containing the vegetable protein is defatted soybean.
(3) The seasoning according to (1) or (2), wherein the microorganism is a filamentous fungus belonging to the genus Aspergillus. (4) The seasoning of (3), wherein the microorganism is Aspergillus oryzae and / or Aspergillus soya.
(5) The following steps:
(I) a step of inoculating a raw material containing plant protein with a microorganism having the ability to hydrolyze the protein to produce a solid koji; and
(Ii) a step of adding a charged solution to the obtained solid koji so as to have a salt concentration such that the hydrolysis reaction of the protein is not inhibited and forming moromi, and fermenting the moromi to hydrolyze the protein. ,
A method for producing a seasoning comprising:
In the step (i), 10 ⁸ to 10 ¹¹ cells / g · raw lactic acid bacteria are added to the raw material, and if necessary, in the step (ii), 10 ⁸ to 10 ¹¹ cells / g · Add to moromi,
The seasoning has an amino acid conversion rate of 65% or more, an isobutyl alcohol concentration of 0.1 mg / g · nitrogen or less, a normal butyl alcohol concentration of 0.25 mg / g · nitrogen or less, and an isoamyl alcohol concentration of 0.5 mg / g · A method characterized by being not more than nitrogen and having an acetic acid concentration of not more than 100 mg / g · nitrogen.
(6) The method according to (5), wherein the salt concentration of the moromi in the step (ii) is 5% by weight or less.
(7) The method according to (5) or (6), wherein the raw material containing the vegetable protein is defatted soybean.
(8) The method according to (7), wherein the defatted soybean is heat-denatured and expanded to an aqueous nitrogen index (NSI) of 8 to 20 by an extruder treatment.
(9) The method according to any one of (5) to (8), wherein the step (ii) is performed at 5 to 45 ° C. for 40 to 144 hours.
(10) The method according to any one of (5) to (9), wherein the pH of the moromi in the step (ii) is 4 to 10.
(11) The method according to any of (5) to (10), wherein in the step (ii), the moromi headspace is replaced with nitrogen having a volume of 2 to 10 times the headspace volume.
(12) The method according to (11), wherein the nitrogen replacement amount is 5 to 8 times the headspace volume.
(13) The method according to any one of (5) to (12), wherein the microorganism capable of hydrolyzing the protein is a filamentous fungus belonging to the genus Aspergillus.
(14) The method according to (13), wherein the microorganism capable of hydrolyzing the protein is Aspergillus oryzae and / or Aspergillus soya.
(15) The method according to any one of (5) to (14), wherein the lactic acid bacterium is Lactococcus lactis.

  According to the present invention, a seasoning containing no salt or having a low salt content, a high amino acid conversion rate, and reduced isobutyl alcohol, normal butyl alcohol, isoamyl alcohol, and acetic acid can be produced.

Hereinafter, the present invention will be described in detail.
The seasoning of the present invention is a seasoning obtained by allowing a microorganism having the ability to hydrolyze a protein to act on a raw material containing a vegetable protein, and has an amino acid conversion rate of 65% or more and an isobutyl alcohol concentration of 0.1 mg / g / nitrogen or less, normal butyl alcohol concentration is 0.25 mg / g / nitrogen or less, isoamyl alcohol concentration is 0.5 mg / g / nitrogen or less, and acetic acid concentration is 100 mg / g / nitrogen or less. It is a seasoning.

  The amino acid conversion is preferably 80% or more, more preferably 85% or more. The isobutyl alcohol concentration is preferably 0.08 mg / g · nitrogen or less, more preferably 0.06 mg / g / nitrogen or less. The normal butyl alcohol concentration is preferably 0.1 mg / g · nitrogen or less, more preferably 0.05 mg / g · nitrogen or less. The isoamyl alcohol concentration is preferably 0.4 mg / g · nitrogen or less, more preferably 0.3 mg / g · nitrogen or less. Further, the acetic acid concentration is preferably 60 mg / g · nitrogen or less, more preferably 30 mg / g · nitrogen or less.

  The amount of nitrogen can be measured, for example, by the Kehldahl method. The amount of amino acids can be measured by an amino acid analyzer, acetic acid can be measured by an organic acid analyzer, and fragrance components can be measured by gas chromatography.

  In the present invention, the “amino acid conversion ratio” is a ratio of a free amino acid to a total amount of amino acids contained in a decomposition solution.

  The raw material containing the plant protein is not particularly limited as long as it is suitable for food and can be efficiently decomposed into amino acids by a microorganism capable of hydrolyzing the protein, and examples thereof include cereals and beans. More specifically, soybeans, particularly defatted soybeans, may be mentioned. In the present invention, the raw material may be one kind or a mixture of two or more kinds. Particularly suitable as a raw material is defatted soybean, but an appropriate amount of flour or the like may be mixed with defatted soybean.

  As microorganisms having the ability to hydrolyze proteins, plant proteins can be hydrolyzed to an amino acid conversion rate of 65% or more, and are suitable for food production, and are proteases such as proteases and peptidases. Microorganisms that secrete extracellularly are preferred. Examples of such a microorganism include microorganisms belonging to the genus Aspergillus, the genus Rhizopus, the genus Mucor, the genus Monascus, and the like. Among these, the genus Aspergillus is preferable, and specifically, Aspergillus oryzae (A. oryzae), Aspergillus sojae (A. sojae), Aspergillus awamori (A. awamori), and Aspergillus nidulans (A. nidulans) And Aspergillus niger. Among these microorganisms, Aspergillus oryzae and Aspergillus soya are particularly preferred.

  Since the seasoning of the present invention as described above has an amino acid conversion ratio of 65% or more, the taste titer of amino acids is higher than that of soy sauce. In addition, the concentrations of isobutyl alcohol, normal butyl alcohol, and isoamyl alcohol, which are aroma components of soy sauce, are lower than those of conventional soy sauce. Furthermore, since the acetic acid concentration is low, the seasoning of the present invention is different from that of conventional soy sauce or deodorized soy sauce (Japanese Patent No. 2862719) in which aroma components are reduced by blowing nitrogen gas. Umami and richness can be imparted without masking the taste and flavor of the material. Further, since salt is not contained or its content is small, it can be suitably used for foods in which the usage of salt is restricted. Further, if necessary, an appropriate amount of salt can be added to the seasoning of the present invention or a food using the same to obtain a desired salt concentration.

  Next, a method for producing the seasoning of the present invention will be described. The seasoning of the present invention is produced, for example, by the following steps.

(I) A step of producing a koji by inoculating a raw material containing a plant protein with a microorganism capable of hydrolyzing a protein (a koji making step).
(Ii) a step of adding a prepared solution to the obtained koji so as to have a salt concentration such that the hydrolysis reaction of the protein is not inhibited and forming moromi, and fermenting the moromi to hydrolyze soybean protein ( Fermentation process).

First, the koji making process will be described.
The raw material containing the plant protein is not particularly limited as long as it is suitable for food and can be efficiently decomposed into amino acids by a microorganism capable of hydrolyzing the protein, and examples thereof include cereals and beans. More specifically, soybeans, particularly defatted soybeans, may be mentioned. In the present invention, the raw material may be one kind or a mixture of two or more kinds. Particularly suitable as a raw material is defatted soybean, but an appropriate amount of flour or the like may be mixed with defatted soybean.

  It is preferable that the defatted soybeans be denatured by heating with an extruder and then dried to obtain dried puffed beans. By doing so, microorganisms having the ability to hydrolyze proteins can easily enter the inside of soybean, and the water content of the raw material can be easily adjusted to 35 to 45% suitable for the growth of Aspergillus. Furthermore, inoculation of a large amount of lactic acid bacteria becomes possible. The heat-denatured expansion is preferably performed so that the water-soluble nitrogen index (NSI) is 8 to 20.

  A koji is prepared by inoculating a raw material as described above with a microorganism capable of hydrolyzing proteins. In the present invention, as the koji, both solid koji and liquid koji can be applied, but solid koji is more protease and peptidase-producing enzymes such as proteases and peptidases produced by the microorganism, and the amino acid conversion is larger. Solid koji is preferred because of its high rate.

In the present invention, at least in the koji-making step of the koji-making step and the fermentation step, 10 ⁸ to 10 ¹¹ cells / g · raw material lactic acid bacterium is added to the raw material. As described later, in the present invention, the solid koji is hydrolyzed at a salt concentration that does not inhibit the hydrolysis reaction of the koji mold, for example, a salt concentration of 5% by weight or less. The term "to the extent that the hydrolysis reaction is not inhibited" includes the case where the hydrolysis reaction is not substantially inhibited and the inhibition is small enough not to impair the effects of the present invention. Specifically, when the degree of amino acid conversion is 65% or more, preferably 80% or more, more preferably 85% or more, it can be said that the hydrolysis reaction is not inhibited. In a conventional soy sauce koji, there are 10 ⁶ to 10 ¹⁰ bacteria / g · koji bacteria at the stage of dekoji, and if such koji is mixed with the charging solution at a low salt concentration, it will spoil in a few hours. Resulting in. In particular, when only defatted soybeans are used as the raw material, the moisture content after steaming becomes very high, 50 to 60%. Therefore, microorganisms such as natto bacteria are better than soy sauce koji made by mixing dry wheat in equal amounts. Pollution is more likely to occur. Therefore, in the present invention, by inoculating lactic acid bacteria into the raw material and the koji, the growth of the contaminating bacteria is suppressed, and the koji is protected from abnormal fermentation and decay caused by the proliferation of the contaminating bacteria.

In the present invention, koji making can be carried out in the same manner as koji making in the production of ordinary soy sauce, except that lactic acid bacteria are inoculated. Specifically, a protein raw material is mixed with water, lactic acid bacteria and seed koji. The water is preferably added in an amount of 35 to 45% by weight, preferably 37 to 42% by weight, based on the total amount of the mixture. The lactic acid bacteria are inoculated so as to have a concentration of 10 ⁸ to 10 ¹¹ cells / g · raw material, preferably 10 ⁹ to 10 ¹⁰ cells / g · raw material. In particular, by inoculating lactic acid bacteria in the above range at the initial stage of koji making, it is possible to maintain a dominant flora in the koji and eliminate the room for other bacteria to grow.

  Lactic acid bacteria are not particularly limited as long as they do not substantially inhibit the activity of microorganisms having the ability to hydrolyze proteins and can suppress the growth of undesired contaminating bacteria. Examples include bacteria belonging to the genus Lactobacillus and the genus Lactococcus. Among these, the genus Lactococcus is preferred, and more specifically, Lactococcus lactis (L. lactis) is mentioned.

Water and lactic acid bacteria can be mixed with raw materials as a culture solution of lactic acid bacteria. Specifically, for example, a lactic acid bacterium culture solution is sprayed on the raw material that has been denatured by heating. The culture solution of lactic acid bacteria preferably contains 10 ⁸ to 10 ¹¹ , preferably 10 ⁹ to 10 ¹⁰ cells / mL. The number of lactic acid bacteria can be measured by counting under a microscope or by counting cfu (colony forming unit) in an agar medium suitable for growth.

The seed koji is usually added so that the number of spores is 10 ⁶ to 10 ⁷ / g · raw material. The number of spores of seed koji can be measured in the same manner as the number of lactic acid bacteria.
The koji making process is usually performed by allowing to stand at 22 to 40 ° C., preferably 28 to 35 ° C., for 24 to 72 hours, preferably 38 to 60 hours. Care (mixing) may be performed 18 to 28 hours after the start of the koji making.

Next, the fermentation step will be described.
The koji prepared as described above is mixed with a preparation liquid to form moromi, and the moromi is fermented to hydrolyze soybean protein. In the present invention, usually, salt is not added to the preparation liquid or moromi, and the salt concentration is preferably 5% by weight or less, more preferably 2% by weight or less, based on the total amount of moromi. In addition, the moromi may contain a small amount of salt derived from a lactic acid bacteria culture solution or the like.

In the fermentation step, it is not essential to add lactic acid bacteria to moromi, but it is preferable. When lactic acid bacteria are added to moromi, lactic acid bacteria are inoculated so as to have a concentration of 10 ⁸ to 10 ¹¹ cells / g · moromi, preferably 10 ⁹ to 10 ¹⁰ cells / g · moromi. By increasing the number of lactic acid bacteria from the initial stage of the fermentation process, it is possible to prevent the lactic acid bacteria from further growing and assimilating the amino acid generated by hydrolysis.

It is desirable to add 1.5 to 5 times, preferably 2 to 4 times the amount of the charged liquid to the weight of the koji. The feed solution and the lactic acid bacteria can be mixed with the raw materials as a culture solution of the lactic acid bacteria. In such a case, the culture solution of lactic acid bacteria preferably contains 10 ⁹ to 10 ¹⁰ cells / mL.

  The fermentation step is performed at a temperature at which lactic acid bacteria can grow, specifically, usually at 5 to 45 ° C, preferably 30 to 37 ° C, for 40 to 144 hours, preferably 48 to 96 hours. The pH of the moromi is preferably adjusted to 4 to 10, more preferably 5 to 7.

  Further, in the fermentation step, it is desirable to replace the head space of the moromi mash with nitrogen gas. As a result, propagation of aerobic contaminants can be suppressed. The degree of nitrogen replacement can be determined, for example, by replacing the headspace volume of the tank containing moromi with nitrogen gas in an amount of 2 to 10 times, preferably 5 to 8 times the amount of nitrogen gas, and sealing the tank.

  By performing the fermentation step as described above, the growth or proliferation of contaminating bacteria can be prevented. In addition, since yeasts involved in the production of normal soy sauce hardly grow, soy sauce aroma is considered to decrease.

  After the completion of the fermentation step, the same treatment as that of ordinary soy sauce can be performed. For example, after the fermented moromi is filtered to remove solids, it is sterilized at 60C to 120C. Alternatively, the moromi may be sterilized and then filtered. Further, the obtained moromi can be used as a raw material for other fermented seasonings or fermented foods.

The present invention will be more specifically described by the following examples. The present invention is not limited by this embodiment.
In this example, “TN” represents nitrogen, and “gTN” represents gram · nitrogen.

<Example 1>
To 360 g of puffed defatted soybean (Protein TY, NSI15 manufactured by Ajinomoto Co., Inc.), 180 mL of a lactic acid bacteria culture solution (L. lactis NBRC12007) containing 10 ⁹ to 10 ¹⁰ cells / mL and adjusted to pH 6.3 was added. Further, after mixing spores of koji mold (A. oryzae JCM2231) at 2 × 10 ⁶ / g · raw material, koji making was carried out at 30 to 32 ° C. for 48 hours according to a conventional method. The koji was mixed 18 and 25 hours after the temperature of the koji exceeded 32 ° C. The water content of the mixture was 37%.
The NBRC12007 strain was deposited with the Fermentation Research Institute (IFO) as IFO12007. However, the preservation of microorganisms by IFO was carried out by the National Institute of Technology and Evaluation, National Institute of Technology and Evaluation, Genetic Resource Division (NBRC) (postal code 292- 0818 2-5-8, Kazusa-Kamazu, Kisarazu-shi, Chiba Prefecture, and is stored as NBRC12007 strain. That is, the NBRC12007 strain is the same strain as the IFO12007 strain. The JCM2231 strain is stored in the RIKEN Microorganism Strain Preservation Facility (JCM) (zip code 351-0198, 2-1 Hirosawa, Wako-shi, Saitama). Both strains can be ordered from NBRC or JCM.

500 g of the obtained koji was added to 2 L of a lactic acid bacterium culture solution (L. lactis NBRC12007 (former IFO12007)) containing 10 ⁹ to 10 ¹⁰ cells / mL and adjusted to pH 6.3, and passed through a pinch cock. It was charged into a pressure-resistant bottle that can shut off the outside air and the inside of the container. After injecting nitrogen gas in an amount equivalent to 5 times the headspace at normal pressure into the head space of the container, the pinch cock is closed, the container is closed, and the container is sealed at 35 ° C. for 24 hours, 48 hours, and 96 hours in an incubator. By leaving still for 144 hours and 240 hours, the defatted soybean protein was hydrolyzed with koji.

  After the hydrolyzed mixture was pressed to prepare a hydrolyzed liquid, the liquid was treated at 80 ° C. for 30 minutes to sterilize it. After incubating this overnight at 4 ° C., the solid content was removed by filtration. To 2 L of the obtained clear liquid, 20 g of activated carbon (activated carbon BA: manufactured by Ajinomoto Fine Techno Co., Ltd.) was added, and the mixture was incubated at 50 ° C. for 30 minutes with constant stirring to perform deodorization and decolorization. Finally, the solution was filtered to remove the activated carbon, and a seasoning solution as a final product was obtained.

  With respect to the seasoning liquid obtained by the above operations, analysis of nitrogen (TN) by the Kjeldahl method, measurement of amino acids (total amino acids) by the amino acid analyzer (Hitachi L-8000), and measurement of various organic compounds by the organic acid analyzer (Hitachi L-7000) Measurement of acid concentration, measurement of various sugar concentrations using a sugar analyzer (Hitachi L-6000), analysis of flavor components by gas chromatography, measurement of pH, and sensory evaluation test using a simple solution system with a sensory evaluation panel consisting of 10 persons Was carried out.

  As a method of sensory evaluation in a simple solution system, each sample was diluted and supplemented so that the nitrogen concentration (TN) = 0.1% and the salt concentration = 1.0%. An evaluation was performed.

  Table 1 shows the results of the component analysis of the prototype at each decomposition time, the results of the sensory evaluation in a simple solution system, and those of a commercially available concentrated soy sauce.

  As a result, by decomposing for 48 hours or more, the amino acid conversion rate reaches 80% or more, and isobutyl alcohol (iba), n-butyl alcohol (nba), isoamyl alcohol (iaa), acetic acid and the like characteristic of soy sauce are obtained. It was found that almost no odor component was contained. The taste of the seasoning solution obtained in the present invention is characterized by a strong umami taste and taste, and a thick taste, and was much closer to the characteristics of the acid-decomposed amino acid solution than soy sauce. In particular, the seasoning liquid having a decomposition time of 48 hours to 144 hours showed strong umami, taste, and thickness. In the case of degradation for 48 hours, a little of the peptide remains, so that the taste becomes “tasteful”.

  As for the odor, no strong flavor peculiar to acid-decomposed amino acids or soy sauce was felt, and only a slight cereal odor was found. There was almost no difference due to the decomposition time. Therefore, in the following examples, the decomposition time was set to 48 hours to 144 hours.

<Example 2>
In the hydrolysis of solid koji, the relationship between the hydrolysis temperature, the production stability, and the taste of the obtained seasoning liquid was examined. The hydrolysis temperature was performed at 30, 35, and 37 ° C. The decomposition time was 96 hours.

500 g of solid koji obtained by the method described in Example 1 was added to 2 L of a lactic acid bacterium culture solution (L. lactis NBRC12007 (former IFO12007)) containing 10 ⁹ to 10 ¹⁰ cells / mL and adjusted to pH 6.3. It was added to a pressure-resistant bottle through which the outside air and the inside of the container could be shut off via a pinch cock. After injecting nitrogen gas in an amount 5 times the headspace volume into the head space of the container, the pinch cock was closed to seal the inside of the container, and hydrolysis was carried out in the incubator at each of the above temperatures for 96 hours. Thereafter, the decomposition product was treated by the method described in Example 1 to obtain a seasoning liquid.

  Table 2 compares the results of component analysis, the results of sensory evaluation in a simple solution system, and the results of microbial analysis after completion of the hydrolysis, with the decomposition temperature being varied.

  As a result, the amino acid conversion ratio did not differ greatly at a decomposition temperature of 30 to 37 ° C.

  Therefore, in the following Examples, the hydrolysis temperature was set to 30 to 37 ° C. in consideration of both the amino acid conversion rate and the bacteriostaticity.

<Example 3>
In Examples 1 and 2, the entire decomposition system was made anaerobic by replacing the headspace above the tank with nitrogen gas during hydrolysis, and the growth of obligate aerobic bacteria including Bacillus subtilis was suppressed. In this example, the necessity for bacteriostasis during hydrolysis was examined.

Lactic acid bacterium culture solution (L. lactis NBRC12007 (former IFO12007)) containing 10 ⁹ to 10 ¹⁰ cells / mL in 360 g of puffed defatted soybean (Protein TY, NSI 15 manufactured by Ajinomoto Co., Inc.) and adjusted to pH 6.3 Was added thereto, and spores of Aspergillus oryzae (A. oryzae JCM2231) were mixed at a rate of 2 × 10 ⁶ spores / g as a raw material, followed by koji making at 30 to 32 ° C. for 48 hours according to a conventional method. The water content of the mixture was 37%. Next, the koji was mixed after 18 and 25 hours in the same manner as in the conventional method for producing soy sauce koji.

500 g of the obtained koji was added to 2 L of a lactic acid bacterium culture solution (L. lactis IFO12007 (former IFO12007)) containing 10 ⁹ cells / mL and adjusted to pH 6.3. It was put into a pressure bottle whose inside can be shut off. The number of lactic acid bacteria was adjusted by diluting a lactic acid bacteria culture solution containing 10 ¹⁰ cells / mL of lactic acid bacteria with sterilized water. Nitrogen gas was injected into the head space of the container, the atmosphere was replaced with nitrogen gas having a volume five times the headspace volume, the pinch cock was closed, the inside of the container was closed, and hydrolysis was performed at 35 ° C. for 48 hours in an incubator. As a control, hydrolysis was performed at 35 ° C. for 96 hours in an incubator without replacing with nitrogen gas. Thereafter, the decomposition product was treated by the method described in Example 1 to obtain a seasoning liquid.

  Under the above conditions, it was found that unless the headspace was replaced with nitrogen, the koji mold covered the moromi surface, and contaminants such as cocci and Bacillus proliferated thereon. Therefore, it was found that it is desirable to replace the head space of the moromi tank with nitrogen.

<Example 4>
Next, the seasoning was manufactured by changing the lactic acid bacteria species used.
Lactococcus lactis (L. lactis) AJ110212 (FERM BP-08552) was added to a medium consisting of 0.54% yeast extract, 3% glucose, and 0.5% NaCl using a jar fan meter, and pH was adjusted with NaOH. Was maintained at 5.5, and cultivation was carried out for about 18 hours under the conditions of no aeration and a stirring speed of 100 rpm so that the cell concentration was 10 ⁹ to 10 ¹⁰ cells / mL. Based on the Budapest Treaty, on November 19, 2003, the National Institute of Advanced Industrial Science and Technology (AIST), the Patent Organism Depositary Center (1-1, Higashi 1-1, Tsukuba, Ibaraki, 305-8566 Japan) Deposited internationally and given accession number FERM BP-08552.
The lactic acid bacterium culture solution (L. lactis AJ110212 (FERM BP-F) adjusted to pH 6.3 containing 360 to 109 g of puffed defatted soybean (Protein TY, NSI15, manufactured by Ajinomoto Co., Inc.) contains 10 ⁹ to 10 ¹⁰ cells / mL. 08552)) was added in an amount of 180 mL. Furthermore, spores of Aspergillus oryzae (A. oryzae JCM2231) were mixed at a rate of 2 × 10 ⁶ / g · raw material, and koji was produced at 30 to 32 ° C. for 48 hours according to a conventional method. The water content of the mixture was 37%. Next, the koji was mixed after 18 and 25 hours in the same manner as in the conventional method for producing soy sauce koji.

500 g of the obtained koji was added to 2 L of a lactic acid bacterium culture solution (culture solution of L. lactis AJ110212 (FERM BP-08552) described above) containing 10 ⁹ cells / mL and adjusted to pH 6.3. It was charged into a pressure-resistant bottle that could shut off the outside air and the inside of the container via a cock. The number of lactic acid bacteria was adjusted by diluting a lactic acid bacteria culture solution containing 10 ¹⁰ cells / mL of lactic acid bacteria with sterilized water. Nitrogen gas was injected into the head space of the container, the atmosphere was replaced with nitrogen gas having a volume five times the head space, the pinch cock was closed, the inside of the container was closed, and hydrolysis was performed at 35 ° C. for 96 hours in an incubator. Thereafter, the decomposition product was treated by the method described in Example 1 to obtain a seasoning liquid.

  Table 3 shows the results of the component analysis of the seasoning solution, the results of the sensory evaluation using a simple solution system, and the results of the microbial analysis.

  As a result, even when L. lactis AJ110212 (FERM BP-08552) was used as the lactic acid bacterium species, the bacteriostatic property of the decomposed solution was ensured, and the sensory evaluation and analysis values were almost the same as the results of Example 1. Was. As described above, the seasoning liquid obtained in the present invention could be produced even when the lactic acid bacteria species was changed.

<Example 5>
Since the seasoning liquid of the present invention contains only a small amount of isobutyl alcohol (iba), n-butyl alcohol (nba), or isoamyl alcohol (iaa), when mixed with a soup, it has a good taste without masking the soup flavor. We thought that we could make noodle soup. Therefore, the soy sauce category was replaced with the seasoning liquid of the present invention to produce soup soup and its sensory evaluation was performed.

  In the combination of soy sauce, bonito flakes (Aramoto flakes, dead flakes), sugar, mirin, salt, L-glutamic acid-sodium salt and mentsuyu as a main component, the soy sauce category was replaced with the following seasonings.

(I) Seasoning solution obtained by the present invention (prepared under the conditions of 35 ° C. and 96 hours in Example 1; hereinafter the same)
(II) Commercial concentrated soy sauce (III) Deodorized soy sauce (prepared by the method described in Japanese Patent No. 2872619)
(IV) Seasoning solution obtained by adding acetic acid to the seasoning solution obtained in the present invention in an amount equal to the amount contained in commercially available concentrated soy sauce.

  Noodle soup using each of the above seasonings was mixed with the following composition to prepare noodle soup, and each was subjected to a sensory evaluation.

Table 3 shows the concentrations of iba, nba, iaaa, and acetic acid contained in each noodle soup.
The above four types of noodle soup were evaluated by a special taste panel (n = 4), and ranked from those with strong soup flavor and smell, and the overall score was given by the ranking method. That is, the first place was 5 points, the second place was 4 points, the third place was 3 points, the fourth place was 2 points, and the fifth place was 1 point, and the total score of the four panels was used as the overall score. The results are shown in Table 4.

  As a result, it was found that when the seasoning liquid of the present invention was used in place of soy sauce, soup was the most dashi flavor and smell. In addition, it was found that as a component for masking the feeling of noodle soup and smoking, the influence of acetic acid was larger than that of iaa, iba, and nba disclosed in Japanese Patent No. 2862719. Therefore, it is presumed that the seasoning liquid of the present invention does not contain acetic acid, thereby having an effect of utilizing the flavor of the material.

Claims (15)

A seasoning obtained by allowing a microorganism having the ability to decompose a protein to act on a raw material containing a vegetable protein, wherein the amino acid conversion rate is 65% or more, the isobutyl alcohol concentration is 0.1 mg / g · nitrogen or less, and normal butyl alcohol is used. A seasoning having a concentration of 0.25 mg / g / nitrogen or less, an isoamyl alcohol concentration of 0.5 mg / g / nitrogen or less, and an acetic acid concentration of 100 mg / g / nitrogen or less. The seasoning according to claim 1, wherein the raw material containing the vegetable protein is defatted soybean. The seasoning according to claim 1 or 2, wherein the microorganism is a filamentous fungus belonging to the genus Aspergillus. The seasoning according to claim 3, wherein the microorganism is Aspergillus oryzae and / or Aspergillus soya. The following steps:
(I) a step of inoculating a raw material containing plant protein with a microorganism having the ability to hydrolyze the protein to produce a solid koji; and
(Ii) a step of adding a charged solution to the obtained solid koji so as to have a salt concentration such that the hydrolysis reaction of the protein is not inhibited and forming moromi, and fermenting the moromi to hydrolyze the protein. ,
A method for producing a seasoning comprising:
In the step (i), 10 ⁸ to 10 ¹¹ cells / g · raw lactic acid bacteria are added to the raw material, and if necessary, in the step (ii), 10 ⁸ to 10 ¹¹ cells / g · Add to moromi,
The seasoning has an amino acid conversion rate of 65% or more, an isobutyl alcohol concentration of 0.1 mg / g · nitrogen or less, a normal butyl alcohol concentration of 0.25 mg / g · nitrogen or less, and an isoamyl alcohol concentration of 0.5 mg / g · A method characterized by being not more than nitrogen and having an acetic acid concentration of not more than 100 mg / g · nitrogen. The method according to claim 5, wherein the salt concentration of the moromi in the step (ii) is 5% by weight or less. The method according to claim 5, wherein the raw material containing the vegetable protein is defatted soybean. The method according to claim 7, wherein the defatted soybean has been heat-denatured and expanded to an aqueous nitrogen index (NSI) of 8 to 20 by an extruder treatment. The method according to any one of claims 5 to 8, wherein the step (ii) is performed at 5 to 45 ° C for 40 to 144 hours. The method according to any one of claims 5 to 9, wherein the pH of the moromi in the step (ii) is 4 to 10. The method according to any one of claims 5 to 10, wherein in step (ii), the moromi headspace is replaced with nitrogen having a headspace volume of 2 to 10 times. The method according to claim 11, wherein the nitrogen replacement is performed at 5 to 8 times the headspace volume. The method according to any one of claims 5 to 12, wherein the microorganism having the ability to hydrolyze the protein is a filamentous fungus belonging to the genus Aspergillus. The method according to claim 13, wherein the microorganism capable of hydrolyzing the protein is Aspergillus oryzae and / or Aspergillus soya. The method according to any one of claims 5 to 14, wherein the lactic acid bacterium is Lactococcus lactis.