JP5306768B2 - Method for producing miso containing ornithine - Google Patents

Description

  The present invention relates to a method for producing miso containing ornithine.

  Ornithine, a kind of amino acid, exists in the body as a free amino acid that does not constitute a protein, and is known to be involved in ammonia detoxification as an important component of the ornithine cycle of the liver. In addition to these liver function improving functions, ornithine is known to have various physiological functions such as growth hormone secretion promoting action, immune function improving action, muscle strengthening action, basal metabolism improving action, and fatigue improving action. ing.

  However, since ornithine is not so much contained in general foods, it is difficult to actively take ornithine in a normal diet. Among them, rainbow trout is relatively abundant, and it has been said that swordfish juice is good for the liver since ancient times. However, even the rainbow trout, which is said to be relatively large, has an ornithine content of about 10 to 15 mg per 100 g.

  For this reason, a method for increasing ornithine content in foods and a method for industrially producing ornithine by microorganisms have been proposed. For example, a method of promoting ornithine synthesis by a stress reaction by applying a cooling load of 0 ° C. or less to a live rainbow trout is known (Patent Document 1). However, since this method can be applied only to live mussel, it is difficult to say that ornithine can be taken in large quantities at low cost.

  In the method of industrially producing ornithine using microorganisms, a method of producing using various strains such as a method of producing and accumulating in a culture medium using an auxotrophic mutant of the genus Brevibacterium (Patent Document 2) Has been proposed. However, in order to ingest them as food, a purification process is required, and it is necessary to add ornithine to the general food later. In addition, since unfamiliar bacterial species are used as food, there is an image resistance to daily consumption as general food.

  Patent Document 3 discloses a method for obtaining a composition or food containing GABA and ornithine at the same time by inoculating a lactic acid bacterium having both γ-aminobutyric acid (GABA) -producing ability and ornithine-producing ability into a culture medium or food material. ing.

  However, in this method, the precursor glutamic acid or a salt thereof and arginine or a salt thereof must be simultaneously present in a food material capable of growing lactic acid bacteria such as a medium, vegetables, fruits and the like. Therefore, treatments such as enzyme treatment of food materials, addition of protein acid hydrolyzate and yeast extract are necessary. A food obtained by such a special production method cannot be equated with a general food obtained by cooking foods. Moreover, since glutamic acid, which is the main ingredient of miso umami, is consumed, there is a problem that umami is reduced.

In particular, Lactobacillus brevis used in the method described in Patent Document 3 is generally poor in salt resistance and alcohol resistance. Therefore, when ornithine is grown in advance in a predetermined medium and added to food, or when a food material capable of growing lactic acid bacteria is used, the food material needs to be pretreated by heat treatment or the like.
JP 2001-204432 A JP-A-2-283290 JP 2008-17703 A

  The objective of this invention is providing the foodstuff which can ingest a sufficient quantity of ornithine cheaply without a sense of incongruity by producing and containing ornithine in the manufacture process of miso.

  According to one aspect of the present invention, there is provided a method for producing miso containing ornithine, which comprises inoculating a lactic acid bacterium that converts arginine into ornithine when producing miso.

  According to the present invention, by producing and containing ornithine in miso without passing through a special manufacturing process, it is possible to provide a food that can be ingested with a sufficient amount of ornithine at a reasonable price.

  Miso is classified into rice miso, barley miso, bean miso, and the like depending on the koji used, but any of the above-mentioned miso may be used as the target of the present invention. Further, the properties such as the length of the aging period and the shade of the color tone are not questioned.

  A general method for producing miso is a koji-making process in which various koji such as rice koji, wheat koji, and soybean koji are obtained by adding koji molds to cereals such as rice and wheat or steamed beans and culturing them; Steamed cereals or beans such as various kinds of koji, salt, water and other ingredients obtained in the koji making process to obtain a koji mixed material; It consists of a fermentation aging process in which the material is fermented and aged to obtain a brew.

  The lactic acid bacteria used in the present invention have the property of converting arginine to ornithine. It is known that soybeans, rice, and the like used as raw materials for miso contain a relatively large amount of arginine, which is a precursor of ornithine. For example, in the case of soybean, arginine accounts for about 8% of the total amino acids contained, and in the case of rice, it accounts for about 5%. Therefore, when the lactic acid bacterium is used in the production of miso made from a raw material containing a high amount of arginine, a miso rich in ornithine can be obtained. This lactic acid bacterium has been confirmed to be safe for use in food. Furthermore, since it is intended to be used for brewing miso, it must have some salt resistance and alcohol resistance.

  The inventors found Lactobacillus brevis 9E53 (FERM P-21671) from the natural world as a lactic acid strain suitable for the production of such miso. This bacterium was isolated from the leaf of vegetables by a known method of anaerobic culture in an MRS medium, and the one in which ornithine was accumulated in the medium was selected.

  In order to confirm the salt tolerance of the separated lactic acid bacteria, culture and screening were carried out by a known method using an MRS medium whose salt concentration was gradually increased. As a result, a salt stress resistant strain having 7% salt concentration and resistance to about 4% ethyl alcohol was further selected.

The characteristics of Lactobacillus brevis 9E53 used in the present invention are summarized below.

  In general miso brewing, cultured salt-tolerant lactic acid bacteria such as Pediococcus halophilus are added and fermented and matured, or mixed into the miso material as resident bacteria at the stage of preparation. The purpose of this is to eliminate the raw material odor, add sourness to promote salt habituation, and lighten the color of the miso so as to produce a tasty appearance. The lactic acid bacteria have salt resistance that can grow in 20% saline solution.

  However, Lactobacillus brevis 9E53 used in the present invention has poor salt resistance. Excluding sweet potato such as Saikyo miso, the salt concentration of common rice miso and bean miso is 10 to 13% by weight. Therefore, with the salt tolerance of Lactobacillus brevis 9E53 lactic acid bacteria used in the present invention, it is not possible to produce miso paste with a normal salt concentration by the above-mentioned ordinary production method. However, in the case of low-salt miso, miso containing ornithine can be produced according to the normal production method described above. In the process, special processes such as enzyme treatment and addition of additives are not required. Here, the low salt miso means a miso having a salt concentration of 7% by weight or less.

  According to one embodiment of the present invention, characterized in that inoculated with lactic acid bacteria that convert arginine to ornithine in the preparation step or fermentation ripening step, the concentration of sodium chloride in the raw material is 7% by weight or less, containing ornithine A method for producing miso is provided. In the case of low salt miso, even if a lactic acid bacterium with poor salt tolerance is used, it can be produced by a usual method comprising a koji making process, a preparation process and a fermentation aging process. As materials for miso other than lactic acid bacteria, those commonly used in the production of general miso can be used, and the temperature, aging period, etc. are also in accordance with general methods.

  In the said method, the lactic acid bacteria which convert arginine into ornithine should just exist in the miso raw material at any time between a preparation process and a fermentation ripening process. The lactic acid bacterium is preferably Lactobacillus brevis 9E53. In order to prevent acidity of the miso, 2 to 5% by weight of ethanol may be added to the charged raw materials.

  According to the above production method, for example, when produced at a koji ratio of 7 and a salt concentration of 7% by weight, a miso containing about 250 mg of ornithine per 100 g of miso can be obtained.

  As described above, with the salt tolerance of Lactobacillus brevis 9E53 lactic acid bacteria used in the present invention, it is not possible to produce miso paste with a normal salt concentration by the above-mentioned production method. In general, in the case of miso brewing using lactic acid bacteria having poor salt tolerance, the salt must be made into a low salt miso by a known method of adding ethanol, a method of adding a large amount of yeast, a method of reducing the amount of water added. However, if the miso ball-like manufacturing method shown below is employed, it is possible to obtain a miso salt having a normal salt concentration of about 13% by weight.

  According to one embodiment of the present invention, the step of mixing steamed soybeans and koji, inoculating the resulting mixture with lactic acid bacteria that convert arginine to ornithine to form a miso-tama-like form, A method for producing ornithine-containing miso comprising the steps of growing and acting the lactic acid bacterium and the step of fermenting and ripening the obtained ornithine-containing raw material is provided.

  When manufacturing bean miso such as Hatcho miso, it is often manufactured by a method called a miso ball manufacturing method. The general method of making miso-tama is to make steamed soybeans into spheroids with a miso-dama making machine to make miso-dama, grow lactic acid bacteria in an anaerobic internal environment, Sprinkle to get miso onion. The obtained miso onion is crushed and mixed with other ingredients and fermented and aged.

  On the other hand, the said embodiment of this invention inoculates ornithine production lactic acid bacteria in the mixture of steamed soybean and koji, and shape | molds it in a miso-tama-like form. By premixing the koji, the koji mold enzyme decomposes the protein and sugar of soybean and rice, and the released arginine is converted to ornithine by the ornithine-producing lactic acid bacteria. In addition, ornithine production is further promoted because energy is supplied to the lactic acid bacteria by decomposition of the carbohydrate. Moreover, since the inside of a miso ball becomes anaerobic like the normal miso ball manufacturing method, lactic acid bacteria propagate actively and the produced | generated lactic acid lowers pH, Therefore The growth of miscellaneous bacteria can also be suppressed.

  The koji used in the miso-tama-like manufacturing method of the present invention may be any of rice koji, wheat koji or bean koji.

  The miso ball in the miso ball-like manufacturing method of the present invention does not indicate only a spherical or square steamed soybean lump of about several centimeters produced by a general miso ball maker or the like. As long as the outside air can be blocked and the inside is anaerobic and lactic acid bacteria can grow, the tank or container may be filled so that there are no voids.

  In the miso ball-like manufacturing method of the present invention, arginine present in the miso ball is converted to ornithine by causing ornithine-producing lactic acid bacteria to grow and act in the miso ball. After the ornithine conversion is completed, the miso ball is crushed and then added with a predetermined yeast, salt, seed water, and, in some cases, koji, to make a raw material. Miso containing ornithine is produced by fermenting and ripening this raw material. Ornithine production takes place during the lactic acid bacteria growth phase in the early stage of fermentation in miso balls. Thereafter, the lactic acid bacteria are killed by the lactic acid produced by the lactic acid bacteria, so that ornithine is not produced during fermentation and ripening after the preparation.

  The ornithine-containing miso produced by the method for producing miso balls of the present invention can convert free arginine to ornithine with an efficiency of almost 100% when the salt concentration is usually 10 to 13% by weight. For example, rice miso of koji yield 6 contains about 430 mg of ornithine per 100 g of miso ball at the end of 10 days of lactic acid fermentation. If a higher concentration of ornithine is desired, arginine can be added in advance to the raw material as a substrate.

The inoculation amount of lactic acid bacteria is preferably set so as to be about 10 ⁵ to 10 ⁸ cells per 1 g of raw materials such as steamed soybeans and strawberries. The larger the inoculum, the shorter the time required for conversion. The time required for ornithine conversion is not particularly limited, but is preferably about 1 to 2 weeks.

  The temperature during the growth of lactic acid bacteria is not particularly limited, but the lactic acid bacteria can be grown in a state adjusted to room temperature or about 30 ° C. Since the amount of ornithine produced can be confirmed by amino acid analysis by HPLC, it is possible to optimize the fermentation conditions such as the time required for lactic acid fermentation and the temperature based on the results.

  In the amino acid analysis by HPLC, 5 g of a sample is dissolved in 45 g of distilled water, stirred for 30 minutes, diluted 10-fold with 0.2N hydrochloric acid, extracted and filtered with stirring, and then subjected to HPLC analysis. The ornithine once produced is stable and does not decrease during the subsequent fermentation and ripening period.

  Lactobacillus brevis 9E53 used in the present invention has an effect of converting arginine to ornithine, but does not act on glutamic acid, which is a major umami component of miso. Therefore, a miso rich in ornithine can be obtained while retaining the umami component. Moreover, since special processes, such as an enzyme treatment, are not required, a general consumer can also take in daily without resistance like normal miso.

  Examples of the present invention will be described below.

Example 1: Low salt miso Soybean 6.4 kg was soaked in water for 16 hours, drained, and steamed at 0.7 kg / cm ² for 30 minutes to obtain 12.8 kg of steamed soybeans. In addition, 4.5 kg of polished rice corresponding to rice bran yield 7 was processed by a conventional method to obtain 4.9 kg of rice bran. To the obtained steamed soybean and rice bran, 1.3 kg of sodium chloride and 0.4 kg of ethanol were added so that the salt concentration of miso was 6.5%.

Add Lactobacillus brevis 9E53 culture solution to this raw material, make the number of lactic acid bacteria in the miso raw material 10 ⁷ cells / g, and add and mix cultured yeast to 10 ⁵ cells / g, Crushed with a chopper. This crushed material was filled into a 30-liter plastic container, the surface was covered with a polyethylene sheet, and fermented and aged by switching halfway at 25 to 35 ° C. for 70 days.

  The obtained low salt miso having a salt concentration of 6.5% by weight had a slightly sour taste, but had the same flavor and fragrance as ordinary low salt miso, and contained 250 mg of ornithine in 100 g of miso.

Example 2: Miso ball-like manufacturing method 5 kg of raw soybeans were immersed in water for 16 hours, drained, and steamed at 0.7 kg / cm ² for 30 minutes to obtain 10 kg of steamed soybeans. To this, 3.5 kg of rice bran was added so that the koji ratio would be 7, and the culture solution of Lactobacillus brevis 9E53 was added and mixed to about 10 ⁵ cells / g in the miso ball, and then crushed with a chopper.

  This ground product was filled in a 30-liter plastic container so that there was no gap, and the surface was covered with a polyethylene sheet to form a miso-tama-like form, which was left in a room at 30 ° C. for 1 hour. This miso ball contained 400 mg of ornithine per 100 g. Moreover, free arginine was not detected.

  Subsequently, this miso ball was crushed with a roller, and salt and yeast were added by a conventional method to adjust the salt concentration to 12.5%. This was filled in a 30 liter plastic container so that there was no gap, charged, and the surface was covered with a polyethylene sheet, and the fermentation was matured by switching halfway at 25 to 35 ° C. for 60 days.

  The obtained miso had the same flavor as normal miso, and contained 400 mg of ornithine in 100 g of miso. Moreover, although the obtained miso was somewhat strong in acidity, when used for cooking miso soup, it had a flavor and aroma that were not different from normal miso.

Comparative Example 1 Ornithine Content of Commercial Miso The ornithine content of the commercial miso was measured. Samples include (1) Takeya miso salt Hikaeme (Takeya Co., Ltd.), (2) Hidejuku (registered trademark) Sendai Miso (Sendai Miso Soy Sauce Co., Ltd.), and (3) Mima Kogen Kura 2 years aged (Marukome) Co., Ltd.) was used.

<Measurement method>
5 g of sample was dissolved in 45 g of distilled water and stirred for 30 minutes. The mixture was diluted 10-fold with 0.2N hydrochloric acid, extracted and filtered with stirring, and subjected to HPLC analysis.

<Result>
(1) Takeya miso salt Hikaeme: 4mg ornithine per 100g miso
(3) Mima Kogen Kura 2 years aging: impossible to measure (trace)
Comparative Example 2: Ornithine content when using other lactic acid strains Miso as described in Examples 1 and 2 using a strain of Lactobacillus brevis having no ornithine production ability instead of Lactobacillus brevis 9E53 Manufactured.

  When the obtained miso was similarly analyzed by HPLC, ornithine was not detected.

Claims (4)

A method for producing miso containing ornithine, which comprises inoculating a lactic acid bacterium that converts arginine into ornithine in the preparation step or fermentation ripening step in producing miso. The concentration of salt in the feed narrowing specification is less than 7 wt%, miso method according to claim 1. Mixing steamed soybeans and koji,
Inoculating the resulting mixture with a lactic acid bacterium that converts arginine to ornithine into a miso-tama-like form;
Causing the lactic acid bacteria to grow and act in the miso ball;
The method for producing miso according to claim 1, comprising a step of fermenting and ripening the obtained ornithine-containing raw material.   The method for producing miso according to any one of claims 1 to 3, wherein the lactic acid bacterium is Lactobacillus brevis 9E53 (FERM P-21671).