JP5974254B2 - Miso manufacturing method and miso manufacturing method - Google Patents

Description

The present invention relates to a method for producing miso for miso and a method for producing miso. More particularly, the method of manufacturing miso for koji to seed koji high productivity and neutral protease activity of vitamin B ₂ such koji mold, and a method of manufacturing a strong umami excellent shades obtained miso using the koji . Hereinafter, vitamin B ₂ refers to riboflavin, flavin mononucleotide, and flavin adenine dinucleotide.

Since many of today's commercial miso are packaged and miso is a seasoning food, color and umami are very important in the quality of miso. It is technically difficult to make a brightly colored miso with excellent color tone, which is a common issue for miso manufacturers. A method of adding vitamin B ₂ is known for light-colored miso to improve color tone. Vitamin B ₂ is a yellow substance and is approved for use as a food additive. Regarding the breeding and utilization of Aspergillus oryzae, as for the soy sauce-like flavor addition and color improvement, white soy sauce koji mold breeding (patent document 1), flavin (vitamin B ₂ class) high-producing koji mold for the purpose of nutrition enhancement, etc. (Non-patent document 1, Non-patent document 2) have been reported.

In addition, in relation to the quality of miso and the enzyme activity of koji, the activity of neutral protease correlates with sensory evaluation of color tone, comprehensive evaluation of sensory content, glutamic acid content of umami ingredients, proteolysis rate, protein dissolution rate, etc. It is reported to be high (Non-patent Document 3), and neutral protease is the most important enzyme in the quality of miso.

Vitamin B ₂ is required as a nutrient component in the body for metabolism and respiration of carbohydrates and proteins, red blood cell formation, antibody production, and normal development. It is indispensable for maintaining the normal activity of the thyroid gland and maintaining the normal health of the entire body, including the skin and hair, and symptoms such as stomatitis and dermatitis occur when insufficient.

Japanese Patent No. 3101140

“Research on Enhanced Miso (Part 2) Production of Flavin by Miso Bacteria by UV Irradiation” Fermentation Engineering 30, 363-369 (1952) Author: Masatoshi Mogi et al. "Generation of flavin-accumulating Bacillus and mycological properties of mutant strains" Science and technology of miso "254, 25-30 (1975) Author: Seinosuke Sakuma et al. "Analysis of various enzymes related to miso quality" Journal of the Japan Brewing Association 87,829-837 (1992) Author: Fuminori Harayama et al.

The method of adding vitamin B ₂ to miso is the use as a food additive, which is not suitable for consumers who do not want to add it. Moreover, the addition of vitamin B ₂ does not lead to enhancement of umami. In practice, an increase in the working process for vitamin B ₂ preparation, there are problems of manufacturing cost increase or the like according to the vitamin B _2. As a solution for these problems, the use of microorganisms such as Neisseria gonorrhoeae having properties relating to color improvement can be considered.

Although the miso described in Patent Document 1 has a good color tone, the colorimetric x value is mainly increased, that is, it is mainly reddish, and the yellowing (colorimetric y value) enhancement suitable for light-colored miso is enhanced. Is insufficient. In addition, since a mutant strain of koji mold for soy sauce is used, the resulting miso exhibits a characteristic soy sauce-like flavor, and unlike the conventional general miso-like flavor, a difference in taste tends to occur.

In Non-Patent Document 1, Mogi et al. Created a mutant strain of miso bacteria with enhanced flavin-producing ability for the purpose of enhancing nutrition, and obtained koji and miso with enhanced vitamin B ₂ species. However, the protease activity of miso and the umami of miso are not particularly enhanced. Proteolytic ability (protease activity) is almost the same as the parent strain, and the quality of miso is the same.

In Non-Patent Document 2, Sakuma et al. Created a flavin-accumulating gonococcal mutant. However, the parent strain of the mutant is not a miso koji-derived but a brewery laboratory-owned test strain derived from a sake brewer's koji, which is practically unsuitable for miso brewing. Further, the protease activity of the obtained mutant strain is lower than that of the parent strain. In addition, the trial brewing of miso has not been carried out, and the effect of miso is not known.

In order to solve such problems, it is an object to easily obtain a miso having a general miso-like flavor, excellent color tone, and enhanced umami by breeding from a practical miso for miso and its use.

In order to solve the above-mentioned problems, the present inventor has bred a koji mold having high productivity of vitamin B ₂ and neutral protease activity by mutation treatment using a koji mold for miso that is practically suitable for miso brewing as a parent strain. I thought it would be effective to use it for miso. The parent strain of Aspergillus is characterized by its high productivity of vitamin B ₂ and neutral protease activity by mutation treatment using a strain held by the Nagano Prefectural Industrial Technology Center isolated from miso soup so that it is practically suitable for miso brewing. Selected stocks. The koji obtained by using this koji mold mutant as a seed koji has the characteristics that the color tone is slightly yellowish from cream color, the content of vitamin B ₂ is high, and the neutral protease activity is high. By using the koji, it was found that a miso having a general miso-like flavor, excellent color tone and strong umami can be easily produced, and the present invention was completed.

That is, the present invention has the following outline.
1 Parent stock of Aspergillus oryzae KJ44, a stock preserved by Nagano Prefectural Industrial Technology Center
As a result of the mutation treatment, to obtain a plurality of mutant strains, using the obtained multiple mutant strains,
A mutant strain producing a cocoon having a vitamin B ₂ content of 1.8 μg / g or more in the obtained koji and a neutral protease activity at pH 6.0 of 60 U / g or more was selected.
Method of manufacturing miso, which comprises using a koji obtained by the method of manufacturing miso for koji according to the production method 2 1 miso for koji, characterized in that the koji from grist with Aspergillus oryzae

The miso miso obtained by the production method of the present invention is a Nagano pref.
As a result, it is practically suitable for miso brewing due to the use of a koji mold mutant with a stock preserved by the National Institute of Advanced Industrial Science and Technology as a parent. Also, the color is slightly yellowish, and the content of vitamin B ₂ and the quality of miso It has excellent properties with high neutral protease activity at pH 6.0, the most important enzyme . The miso obtained by the manufacturing method using this miso koji has a vivid and excellent color tone without using vitamin B ₂ which is a food additive. As for the flavor, the umami is enhanced while maintaining a general taste-like taste, the composition (physical properties) becomes soft and smooth, and a miso of high commercial value with excellent color, taste and composition can be obtained. Particularly effective for light-colored miso. In addition, when the miso koji obtained by the production method of the present invention is used as salt koji, the product has a yellowish color and a beautiful color, and when used as a fish or meat pickling floor, the fish and meat are softened quickly. In addition, the umami can be enhanced. Both the miso for miso and miso have increased vitamin B ₂ content, resulting in products with enhanced nutritional value as described in the background art.

Hereinafter, the present invention will be described in detail.

The present invention is characterized in that koji and miso are produced using koji mold having high productivity of vitamin B ₂ and neutral protease activity. The parent strain of Aspergillus belongs to Aspergillus oryzae, and the Nagano Prefectural Industrial Technology Center preserved strain isolated from miso koji can be used. Mutation treatment is performed on the spores of this strain, and the isolated strain is subjected to a koji-making test. As the mutation treatment, any method such as ultraviolet ray irradiation, X-ray irradiation, treatment with a drug such as N-methyl-N′-nitro-N-nitrosoguanidine can be adopted. In the koji-making test, steamed rice is inoculated with a mutant strain and kept at a predetermined temperature for a predetermined time.

By selecting the color of the resulting koji using yellow as an index, analyzing the components of vitamin B ₂ by high-performance liquid chromatography, and measuring the enzyme activity, koji molds with high productivity and neutral protease activity of vitamin B ₂ are selected. Can get. Vitamin B ₂ and neutral protease are measured according to the standard miso analysis method [edited by the National Miso Technical Association (1995)]. Examples of the koji mold selected by the above method include R2 strains in which rice bran produced according to a conventional method has a vitamin B ₂ content of 1.8 μg / g or more and a neutral protease activity of 60 U / g or more. Here, the unit (U) of neutral protease activity uses a milk casein (Merck) solution at pH 6.0 as a substrate, and develops a product such as tyrosine produced by an enzyme reaction at 37 ° C. for 1 hour with a phenol reagent. It was expressed as absorbance at 660 nm. At that time, the time when the absorbance at 660 nm was 0.35 with respect to 50 μg / ml of the tyrosine solution was calculated as the Folin coefficient = 1.0. Finally, the activity was expressed in terms of dry matter weight of straw.

When making koji using the koji mold selected as described above, using cereals as a raw material, for example, in the production of rice koji, inoculating the koji mold as a seed koji into steamed rice obtained by processing the raw rice as usual. Then, using a iron making apparatus or the like, the temperature is maintained for a predetermined time. The color tone of the koji according to the present invention is slightly yellowish from cream color, unlike the white color generally obtained by conventional commercially available koji molds or parent strains. In addition, the content of vitamin B ₂ is 1.8 μg / g or more and the neutral protease activity is 60 U / g or more.

In order to produce miso using the koji of the present invention, ordinary methods may be followed, mixing and charging koji, cereal raw materials such as boiled soybeans, salt, seed water, yeast, lactic acid bacteria, etc. if necessary, Fermentation and aging for a predetermined period at a predetermined temperature. Although the miso according to the present invention is slightly colored as compared with the conventional miso, the colorimetric x value and y value are both high and the color tone is excellent and the color tone is strong. Furthermore, it is a general miso-like flavor and has a high FN (formol nitrogen) and WSN (water-soluble nitrogen), and a strong miso.

Hereinafter, the present invention will be described in more detail with reference to examples.

Aspergillus oryzae KJ44, a stock of Nagano Prefectural Industrial Technology Center, was used as the parent strain of Aspergillus. This strain was cultured in a potato dextrose medium to form spores, and the cells were collected with 0.01% Tween 80, and the spores were collected by filtration through a 3G3 glass filter. 2 × 10 ⁶ / ml spores were subjected to mutation treatment at 30 ° C. for 30 minutes with 2 mg / ml of N-methyl-N′-nitro-N-nitrosoguanidine. The isolated mutant was inoculated into about 100 g of steamed rice in a petri dish and cultured at 35 ° C. for 48 hours or further maintained for up to 5 days. The resulting wrinkles were visually judged using yellow as an index, and R1 and R2 strains having a slightly yellowish tone from the cream color were selected.

For the R1 and R2 strains selected using the color of koji as an index, a koji-making test was conducted to select a strain that is highly practical and suitable for improving the color tone and enhancing the taste of miso. As a control strain, a commercially available miso seed mold BF-1 (Matsunosuke Takaguchi Co., Ltd.) was used. The seed strain of the selected strain was seeded on 3.8 kg of steamed rice that had been treated in accordance with a conventional method, and the resultant was sown at 37 ° C. for 46 hours. The obtained koji was subjected to visual growth and color evaluation, water content measurement, quantitative analysis of vitamin B ₂ by high performance liquid chromatography, neutral protease activity measurement, and the like. The measurement of vitamin B ₂ and neutral protease was in accordance with the standard miso analysis method.

At the time of koji making, R1 grew slower than BF-1, and the scouring circumference was slightly inferior. Unlike the BF-1 bacteria, the color of the cocoon was slightly yellowish. As a result of the slow growth, the water content of the straw was high and the neutral protease activity tended to be low (Table 1). On the other hand, the viability of R2 around the semen was slightly inferior to that of BF-1, but not so much as to be a problem. The color of the cocoon was slightly yellowish like R1. The water content was almost the same as that of BF-1 (Table 1), and the neutral protease showed a high activity of 66 U / g (Table 1). Vitamin B ₂ content of strawberry was 0.79 μg / g of BF-1, R1 was 2.59 μg / g and R2 was 2.06 μg / g, which was about 3 times as much (Table 1). The origin of cocoon yellow is not limited to vitamin B ₂ species, but it is considered to be at least one of the yellow substances.
Based on the results of the above koji-making test, the R2 strain was selected as a koji mold suitable for practical use and having high productivity of vitamin B ₂ and high neutral protease activity.

Example 1
Using the R2 strain selected as described above, a miso-making test was conducted at a miso manufacturing company. As a control, the above-described commercially available miso seed bacilli BF-1 was used. R2 seeds selected from steamed rice treated with 90 kg of domestic round rice as a raw material were seeded as raw material rice, and then kneaded at 36 ° C. for 44 hours using an awning type ventilator. R2 followed the process of making rice mash for general miso, and the viability of semen and the like was almost the same as that of BF-1. The color of the cocoon was clearly different from that of BF-1, and was slightly yellowish. The water content was lower than that of BF-1, suggesting vigorous growth (Table 2). Further, the neutral protease activity of R2 was 108 U / g, and the content of vitamin B ₂ was 2.86 μg / g, which were very high values (Table 2).

Example 2
Using the koji manufactured in Example 1, a miso preparation test was conducted at a miso manufacturing company. The feed setting was 10% cocoon ratio, moisture 47%, salt 12%, total charge about 350 kg, aging at 30 ° C. for 20 days, and then aging at room temperature for 15 days. The general component analysis results of miso after aging were shown (Table 3). Regarding the hue, the Y value of the R2 section was about 2% lower than that of the BF-1 section, and the coloration tended to proceed slightly. On the other hand, in the R2 section, both the x value indicating reddish hue and the y value indicating yellowish hue were remarkably high, and the color tone was excellent. In the light-colored miso having a short ripening as in this example, it is considered that the yellow taste of the koji derived from vitamin B ₂ and the like is greatly involved in the increase in the y value. The effect of neutral protease activity is presumed to increase the x value. Moreover, FN and WSN of R2 section showed a higher value than a control section, and it was suggested that there are many umami components. This is considered to reflect the high neutral protease activity of sputum. In addition, although yeast was added at the time of preparation, almost no alcohol was produced in both sections.

Sensory evaluation of the miso of Example 2 was performed (Table 4). In addition, the score was obtained by a five-point method, and the lower the numerical value, the better the result. The average value of six panelists was shown. R2 ward got high evaluation in color, taste, composition and synthesis compared to BF-1 ward. Although the color of R2 was slightly colored, it had a shimmering and harsh appearance and was clearly an excellent color tone. The taste was salty and the taste was strong and the composition was soft and smooth, both of which were good. High neutral protease activity can be considered to contribute directly or indirectly to color, taste and composition. Although the fragrance was poor in fermented fragrance in both categories, it was in the category of miso-like fragrance.

INDUSTRIAL APPLICATION This invention is utilized for fermented food manufacture, mainly manufacture of miso, and can obtain the product which is excellent in color tone and strong in umami.

Claims (2)

Aspergillus oryzae KJ44, a stock preserved by Nagano Industrial Technology Center
To obtain a plurality of mutant strains, and using the plurality of mutant strains obtained,
A mutant strain producing a cocoon having a vitamin B ₂ content of 1.8 μg / g or more in the obtained koji and a neutral protease activity at pH 6.0 of 60 U / g or more was selected. A method for producing miso koji, characterized in that koji molds are used to make koji from cereal raw materials. A method for producing miso, comprising using the koji obtained by the method for producing miso for miso according to claim 1 .