JP6715170B2

JP6715170B2 - Food manufacturing method

Info

Publication number: JP6715170B2
Application number: JP2016229502A
Authority: JP
Inventors: 剛杉山; 典白土屋; 内田　建; 建内田; 奈津子古賀; 祥子樋口
Original assignee: 有限会社オトコーポレーション
Priority date: 2016-11-25
Filing date: 2016-11-25
Publication date: 2020-07-01
Anticipated expiration: 2036-11-25
Also published as: JP2018082681A

Description

本発明は、食品の製造方法に関する。 The present invention relates to a method for producing food.

納豆は、大豆を原料として、納豆菌により発酵させた食品であり、消化作用や栄養吸収作用を有するプロテアーゼ、抗炎症作用を有するポリアミン、セロトニンを生成するトリプトファン、腸管免疫に関与するグルタミン、ＧＡＢＡ、サポニン、イソフラボン、その他の抗酸化物質など、多くの有効成分を含むことが知られている。また、納豆の発酵過程では、ビタミンＢ２が増加することが知られており、脂肪燃焼、及びダイエット効果を有することが期待されている。
更に、前記納豆菌は、Ｏ−１５７などに対する抗菌物質を産生することが知られており、抗菌剤としての有用性も期待されている。 Natto is a soybean-based material that is fermented with Bacillus natto, a protease having a digestive action and a nutrient absorption action, a polyamine having an anti-inflammatory action, tryptophan that produces serotonin, glutamine involved in intestinal immunity, GABA, It is known to contain many active ingredients such as saponins, isoflavones, and other antioxidants. In addition, it is known that vitamin B2 is increased in the fermentation process of natto, and it is expected to have fat burning and diet effects.
Furthermore, it is known that the Bacillus natto produces an antibacterial substance against O-157 and the like, and is expected to be useful as an antibacterial agent.

上記期待と一致して、納豆市場は拡大傾向にあるが、市場での価格競争を考慮して、付加価値を高めた納豆の開発が求められている。 Consistent with the above expectations, the natto market is expanding, but considering the price competition in the market, the development of natto with increased added value is required.

乳酸菌、酪酸菌及び麹菌は、整腸効果を有することが知られており、古くからヨーグルト、ぬか漬け、日本酒などの食品の発酵に使用されている。 Lactic acid bacteria, butyric acid bacteria, and koji mold are known to have an intestinal regulating effect, and have been used for fermentation of foods such as yogurt, rice bran pickles, and sake since ancient times.

そこで、前記納豆の優れた機能を損なうことなく、前記乳酸菌、酪酸菌、及び麹菌の少なくともいずれかにより付加価値を高めた食品の開発が望まれるが、これまでに、納豆菌の強い抗菌作用のため、納豆菌と乳酸菌等との共存は不可能であることが示されてきた。 Therefore, without impairing the excellent function of the natto, it is desired to develop a food with increased value added by at least one of the lactic acid bacterium, the butyric acid bacterium, and the koji mold, but so far, of the strong antibacterial action of the natto bacterium. Therefore, it has been shown that natto bacteria and lactic acid bacteria cannot coexist.

また、納豆、及び乳酸菌培養液を含む家畜用飼料の製造方法については開示されているが（特許文献１）、これは、米ぬか味噌と、納豆と、乳酸菌培養液と、土着菌が生息する腐葉土と、米ぬかとを混合して発酵させるものであり、納豆の優れた機能を損なうことなく、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかによる付加価値を高めた食品の簡便な製造方法については報告されていない。 Moreover, although a method for producing a feed for livestock containing natto and a lactic acid bacterium culture solution is disclosed (Patent Document 1), this is rice bran miso, natto, a lactic acid bacterium culture solution, and a leaf-leaf soil inhabiting indigenous bacteria. And rice bran are mixed and fermented, and a simple method for producing foods with increased added value of at least one of lactic acid bacteria, butyric acid bacteria, and koji mold is reported without impairing the excellent function of natto. It has not been.

特開２０１０−２２３６８号公報JP, 2010-22368, A

本発明は、従来における前記諸問題を解決し、以下の目的を達成することを課題とする。即ち、本発明は、納豆の優れた機能を損なうことなく、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかにより付加価値を高めた食品の簡便な製造方法を提供することを目的とする。 An object of the present invention is to solve the above-mentioned problems in the related art and achieve the following object. That is, an object of the present invention is to provide a simple method for producing a food product having an added value enhanced by at least one of lactic acid bacteria, butyric acid bacteria, and koji mold, without impairing the excellent function of natto.

本発明者らが、前記目的を達成すべく鋭意研究を重ねた結果、納豆菌含有物と、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかとを混合し、前記乳酸菌、酪酸菌、及び麹菌の少なくともいずれかで前記納豆菌含有物を発酵することにより、納豆の優れた機能を損なうことなく、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかによる付加価値を高めた食品の簡便な製造方法が提供できることを確認した。 The present inventors, as a result of repeated intensive research to achieve the above object, a natto bacteria-containing material, lactic acid bacterium, butyric acid bacterium, and at least one of koji mold are mixed, at least one of the lactic acid bacterium, butyric acid bacterium, and koji mold. By fermenting the natto bacteria-containing material in any one, it is possible to provide a simple method for producing a food product with increased added value by at least one of lactic acid bacteria, butyric acid bacteria, and koji mold, without impairing the excellent function of natto. It was confirmed.

本発明は、本発明者らによる前記知見に基づくものであり、前記課題を解決するための手段としては以下の通りである。即ち、
＜１＞納豆菌含有物と、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかとを混合し、前記乳酸菌、酪酸菌、及び麹菌の少なくともいずれかで前記納豆菌含有物を発酵する発酵工程を含むことを特徴とする食品の製造方法である。
＜２＞前記乳酸菌が、少なくとも２種類の乳酸菌を含む＜１＞に記載の製造方法である。
＜３＞前記食品における前記納豆菌、乳酸菌、酪酸菌、及び麹菌の濃度が、少なくとも以下のいずれかを満たす＜１＞から＜２＞のいずれかに記載の製造方法；
前記納豆菌の濃度が、生菌で、１×１０^８個／ｇ以上である、
前記乳酸菌の濃度が、生菌で、１×１０^８個／ｇ以上である、
前記酪酸菌の濃度が、生菌で、１×１０^８個／ｇ以上である、又は
前記麹菌の濃度が、生菌で、１×１０^８個／ｇ以上である。
＜４＞前記食品におけるプロテアーゼ力価が、１，０００ｕ／ｇ以上である＜１＞から＜３＞のいずれかに記載の製造方法である。
＜５＞前記発酵工程において、炭酸カルシウムを添加する＜１＞から＜４＞のいずれかに記載の製造方法である。
＜６＞前記発酵工程において、ビタミンＣを添加する＜１＞から＜５＞のいずれかに記載の製造方法である。 The present invention is based on the above findings by the present inventors, and means for solving the above problems are as follows. That is,
<1> A fermenting step of mixing the natto bacterium-containing material with at least one of lactic acid bacterium, butyric acid bacterium, and koji mold, and fermenting the natto bacterium containing material with at least one of the lactic acid bacterium, butyric acid bacterium, and koji mold Is a method for producing a food product.
<2> The production method according to <1>, wherein the lactic acid bacterium contains at least two types of lactic acid bacteria.
<3> The production method according to any one of <1> to <2>, in which the concentrations of the Bacillus natto, lactic acid bacterium, butyric acid bacterium, and koji mold in the food satisfy at least one of the following:
The concentration of the Bacillus natto is 1×10 ⁸ viable bacteria/g or more,
The concentration of the lactic acid bacterium is 1×10 ⁸ viable bacteria/g or more,
The concentration of the butyric acid bacteria, in live bacteria, is 1 × 10 ⁸ cells / g or more, or the concentration of the Aspergillus oryzae, in live bacteria, is 1 × 10 ⁸ cells / g or more.
<4> The production method according to any one of <1> to <3>, wherein the protease titer in the food is 1,000 u/g or more.
<5> The production method according to any one of <1> to <4>, wherein calcium carbonate is added in the fermentation step.
<6> The production method according to any one of <1> to <5>, wherein vitamin C is added in the fermentation step.

本発明によると、従来における前記諸問題を解決し、前記目的を達成することができ、納豆の優れた機能を損なうことなく、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかにより付加価値を高めた食品の簡便な製造方法を提供することができる。 According to the present invention, it is possible to solve the above-mentioned problems in the related art, achieve the above-mentioned object, and increase the added value by at least one of lactic acid bacteria, butyric acid bacteria, and koji mold without impairing the excellent function of natto. It is possible to provide a simple method for producing food.

図１は、実施例１−１及び１−２並びに比較例１−１及び１−２で得られた食品の乳酸菌数を示す図である。FIG. 1 is a diagram showing the number of lactic acid bacteria in the foods obtained in Examples 1-1 and 1-2 and Comparative Examples 1-1 and 1-2. 図２は、実施例２−１〜２−３及び比較例２で得られた食品の乳酸菌数を示す図である。FIG. 2 is a diagram showing the number of lactic acid bacteria in the foods obtained in Examples 2-1 to 2-3 and Comparative Example 2. 図３は、実施例２−１〜２−３及び比較例２で得られた食品の納豆菌数を示す図である。FIG. 3 is a diagram showing the numbers of Bacillus natto of the foods obtained in Examples 2-1 to 2-3 and Comparative Example 2. 図４は、実施例３で得られた食品の酪酸菌数及び納豆菌数を示す図である。FIG. 4 is a diagram showing the numbers of butyric acid bacteria and natto bacteria of the food obtained in Example 3. 図５は、実施例４で得られた食品の酪酸菌数を示す図である。FIG. 5 is a diagram showing the number of butyric acid bacteria in the food obtained in Example 4. 図６は、実施例５で得られた食品の写真である。FIG. 6 is a photograph of the food product obtained in Example 5. 図７は、実施例５で得られた食品の顕微鏡写真である。FIG. 7 is a photomicrograph of the food product obtained in Example 5. 図８は、実施例６−１及び６−２で得られた食品の乳酸菌数を示す図である。FIG. 8 is a diagram showing the number of lactic acid bacteria in the foods obtained in Examples 6-1 and 6-2. 図９は、実施例６−１及び６−２で得られた食品の納豆菌数を示す図である。FIG. 9: is a figure which shows the natto bacteria count of the foodstuff obtained in Examples 6-1 and 6-2. 図１０は、実施例７−１及び７−２で得られた食品の乳酸菌数及び納豆菌数を示す図である。FIG. 10 is a diagram showing the numbers of lactic acid bacteria and natto bacteria of the foods obtained in Examples 7-1 and 7-2. 図１１は、実施例８−１及び８−２で得られた食品の乳酸菌数及びｐＨを示す図である。FIG. 11 is a diagram showing the number of lactic acid bacteria and the pH of the foods obtained in Examples 8-1 and 8-2. 図１２は、実施例９で得られた食品の乳酸菌数及びプロテアーゼ力価を示す図である。FIG. 12 is a diagram showing the number of lactic acid bacteria and the protease titer of the food obtained in Example 9.

（食品の製造方法）
本発明の食品の製造方法は、発酵工程を含み、更に必要に応じてその他の工程を含むことができる。 (Food manufacturing method)
The method for producing a food of the present invention includes a fermentation step, and may further include other steps as necessary.

＜発酵工程＞
前記発酵工程は、納豆菌含有物と、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかとを混合し、前記乳酸菌、酪酸菌、及び麹菌の少なくともいずれかで前記納豆菌含有物を発酵する工程である。 <Fermentation process>
The fermentation step is a step of mixing the natto bacterium-containing material with at least one of lactic acid bacterium, butyric acid bacterium, and koji mold, and fermenting the natto bacterium containing material with at least one of the lactic acid bacterium, butyric acid bacterium, and koji mold. ..

＜＜納豆菌含有物＞＞
前記納豆菌含有物としては、納豆を含むものであれば特に制限はなく、目的に応じて適宜選択することができ、更に必要に応じてその他の添加物を加えることができる。 <<<Natto bacterium inclusions>>
The natto bacterium-containing material is not particularly limited as long as it contains natto, and can be appropriately selected according to the purpose, and other additives can be added as necessary.

−納豆−
前記納豆としては、市販品であってもよく、従来公知の手法に基づき、大豆及び納豆菌を用いて製造したものであってもよい。
前記納豆の形態としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、粗挽きした納豆、粗挽きしていない納豆、これらの併用などが挙げられるが、発酵効率の点から、粗挽きした納豆が好ましい。
前記粗挽きした納豆は、前記納豆を市販の挽肉機を用いて粗挽きすることにより得ることができる。
前記納豆菌としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、Ｂａｃｉｌｌｕｓｓｕｂｔｉｌｉｓｎａｔｔｏなどが挙げられる。 -Natto-
The natto may be a commercially available product, or may be produced using soybean and natto bacterium based on a conventionally known method.
The form of the natto is not particularly limited and can be appropriately selected depending on the purpose, and includes, for example, coarsely ground natto, uncooked natto, and a combination thereof, but in terms of fermentation efficiency. Therefore, coarsely ground natto is preferable.
The roughly ground natto can be obtained by roughly grinding the natto using a commercially available meat grinder.
The Bacillus natto is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include Bacillus subtilis natto.

−その他の添加物−
前記その他の添加物としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、米粉などが挙げられる。 -Other additives-
The other additives are appropriately selected depending on the intended purpose without any limitation, and examples thereof include rice flour.

＜＜乳酸菌＞＞
前記乳酸菌としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、Ｌａｃｔｏｂａｃｉｌｌｕｓａｃｉｄｏｐｈｉｌｕｓ、Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍｌｏｎｇｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｃａｓｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｐｌａｎｔａｒｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓａｋｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎｅｓなどが挙げられる。
これらの乳酸菌は、１種単独で使用してもよいし、２種以上を併用してもよいが、発酵効率の点から、少なくとも２種類の乳酸菌を含むことが好ましい。
前記乳酸菌としては、熱や化学物質、エックス線などに高い耐久性を有し、保存性の向上を図ることができる点から、有胞子乳酸菌が好ましい。前記有胞子乳酸菌は、栄養の枯渇、温度や湿度などの変化、老廃物の蓄積など、菌の生存に不利な環境に置かれた場合に芽胞を形成する。前記不利な環境としては、特に制限はなく、目的に応じて適宜選択することができるが、例えば、低湿度（乾燥）環境、高温環境などが挙げられる。
前記Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎｅｓは、有胞子乳酸菌である。
前記乳酸菌は、市販品であってもよい。 << lactic acid bacteria >>
The lactic acid bacterium is not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus lucilis lac, and the like.
These lactic acid bacteria may be used alone or in combination of two or more, but it is preferable to contain at least two kinds of lactic acid bacteria from the viewpoint of fermentation efficiency.
The lactic acid bacterium is preferably a spore-forming lactic acid bacterium, since it has high durability against heat, chemical substances, X-rays, etc. and can improve storage stability. The spore-forming lactic acid bacterium forms spores when placed in an environment that is disadvantageous to the survival of the bacterium, such as depletion of nutrients, changes in temperature and humidity, and accumulation of waste products. The disadvantageous environment is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a low humidity (dry) environment and a high temperature environment.
The Lactobacillus sporogenes are spore lactic acid bacteria.
The lactic acid bacterium may be a commercially available product.

前記乳酸菌の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、乳酸菌０．１〜１０質量部が好ましく、０．３〜５質量部がより好ましく、０．３〜２質量部が更に好ましい。 The amount of the lactic acid bacterium with respect to the natto is not particularly limited and may be appropriately selected according to the purpose, but 0.1 to 10 parts by mass of lactic acid bacterium is preferable with respect to 100 parts by mass of natto, and 0.3 to 5 parts by mass is more preferable, and 0.3 to 2 parts by mass is further preferable.

＜＜酪酸菌＞＞
前記酪酸菌としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、Ｃｌｏｓｔｒｉｄｉｕｍｂｕｔｙｒｉｃｕｍなどが挙げられる。
前記酪酸菌としては、熱や化学物質、エックス線などに高い耐久性を有し、保存性の向上を図ることができる点から、有胞子酪酸菌が好ましい。前記有胞子酪酸菌は、栄養の枯渇、温度や湿度などの変化、老廃物の蓄積など、菌の生存に不利な環境に置かれた場合に芽胞を形成する。前記不利な環境としては、特に制限はなく、目的に応じて適宜選択することができるが、例えば、低湿度（乾燥）環境、高温環境などが挙げられる。
前記Ｃｌｏｓｔｒｉｄｉｕｍｂｕｔｙｒｉｃｕｍは、有胞子酪酸菌である。
前記酪酸菌は、市販品であってもよい。 << Butyric acid bacteria >>
The butyric acid bacterium is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include Clostridium butyricum.
As the butyric acid bacterium, spore-containing butyric acid bacterium is preferable because it has high durability against heat, chemical substances, X-rays, etc. and can improve storage stability. The spore-containing butyric acid bacterium forms spores when placed in an environment that is disadvantageous to the survival of the bacterium, such as depletion of nutrients, changes in temperature and humidity, and accumulation of waste products. The disadvantageous environment is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a low humidity (dry) environment and a high temperature environment.
The Clostridium butyricum is a spore butyric acid bacterium.
The butyric acid bacterium may be a commercially available product.

前記酪酸菌の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、酪酸菌０．１〜１０質量部が好ましく、０．３〜５質量部がより好ましく、０．３〜２質量部が更に好ましい。 The amount of the butyric acid bacterium with respect to the natto is not particularly limited and may be appropriately selected depending on the purpose, but is preferably 0.1 to 10 parts by mass of butyric acid bacterium with respect to 100 parts by mass of natto, and 0.1. 3 to 5 parts by mass is more preferable, and 0.3 to 2 parts by mass is even more preferable.

＜＜麹菌＞＞
前記麹菌としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、黄麹菌、白麹菌、黒麹菌などが挙げられる。
前記麹菌は、種麹であってもよい。
前記種麹としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、清酒用種麹などが挙げられる。
前記麹菌は、市販品であってもよい。 <<Koji mold>>
The koji mold is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include yellow koji mold, white koji mold and black koji mold.
The koji mold may be seed koji.
The seed koji is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include sake koji for sake.
The koji mold may be a commercially available product.

前記麹菌の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、麹菌０．０１〜１０質量部が好ましく、０．１〜５質量部がより好ましく、０．１〜２質量部が更に好ましい。 The amount of the koji mold to the natto is not particularly limited and can be appropriately selected depending on the purpose, but 0.01 to 10 parts by mass of koji mold is preferable with respect to 100 parts by mass of natto, and 5 parts by mass is more preferable, and 0.1 to 2 parts by mass is further preferable.

＜＜発酵＞＞
前記発酵の方法としては、前記納豆菌含有物を、前記乳酸菌、酪酸菌、及び麹菌の少なくともいずれかで発酵できる方法であれば特に制限はなく、目的に応じて適宜選択することができ、例えば、好気発酵、嫌気発酵などが挙げられる。更に必要に応じてその他の添加物を加えて発酵することができる。 <<Fermentation>>
The fermentation method is not particularly limited as long as it is a method in which the natto bacterium-containing material can be fermented with at least one of the lactic acid bacterium, butyric acid bacterium, and koji mold, and can be appropriately selected depending on the purpose, for example, , Aerobic fermentation, anaerobic fermentation and the like. If necessary, other additives can be added for fermentation.

−好気発酵−
前記好気発酵としては、好気条件下での発酵であれば、特に制限はなく、目的に応じて適宜選択することができ、例えば、好気条件下において、培養機内で静置する方法などが挙げられる。
前記好気発酵の発酵温度としては、特に制限はなく、目的に応じて適宜選択することができるが、１０℃〜５０℃が好ましく、２０℃〜４０℃がより好ましく、３０℃〜４０℃が更に好ましい。
前記好気発酵の発酵時間としては、特に制限はなく、目的に応じて適宜選択することができるが、１時間〜７日間が好ましく、１２時間〜９６時間がより好ましく、２４時間〜４８時間が更に好ましい。 -Aerobic fermentation-
The aerobic fermentation is not particularly limited as long as it is fermentation under aerobic conditions, and can be appropriately selected depending on the purpose, for example, under aerobic conditions, a method of leaving it in a culture machine, etc. Are listed.
The fermentation temperature of the aerobic fermentation is appropriately selected depending on the intended purpose without any limitation, but it is preferably 10°C to 50°C, more preferably 20°C to 40°C, and 30°C to 40°C. More preferable.
The fermentation time of the aerobic fermentation is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 1 hour to 7 days, more preferably 12 hours to 96 hours, and 24 hours to 48 hours. More preferable.

−嫌気発酵−
前記嫌気発酵としては、嫌気条件下での発酵であれば、特に制限はなく、目的に応じて適宜選択することができ、例えば、嫌気条件下において、培養機内で静置する方法などが挙げられる。
前記嫌気発酵の発酵温度としては、特に制限はなく、目的に応じて適宜選択することができるが、１０℃〜５０℃が好ましく、２０℃〜４０℃がより好ましく、３０℃〜４０℃が更に好ましい。
前記嫌気発酵の発酵時間としては、特に制限はなく、目的に応じて適宜選択することができるが、１時間〜７日間が好ましく、１２時間〜９６時間がより好ましく、２４時間〜４８時間が更に好ましい。 -Anaerobic fermentation-
The anaerobic fermentation is not particularly limited as long as it is a fermentation under anaerobic conditions, and can be appropriately selected according to the purpose. For example, under anaerobic conditions, a method of standing in a culture machine and the like can be mentioned. ..
The fermentation temperature of the anaerobic fermentation is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 10°C to 50°C, more preferably 20°C to 40°C, and further preferably 30°C to 40°C. preferable.
The fermentation time of the anaerobic fermentation is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 1 hour to 7 days, more preferably 12 hours to 96 hours, further preferably 24 hours to 48 hours. preferable.

−その他の添加物−
前記その他の添加物としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、炭酸カルシウム、ビタミンＣ、クエン酸、ＮａＣｌ溶液、蒸留水などが挙げられる。
これらのその他の添加物は、１種単独で使用してもよいし、２種以上を併用してもよいが、乳酸菌の代謝産物である乳酸の中和の促進、嫌気性発酵効率、納豆菌の増殖抑制などの点から、炭酸カルシウム、又はビタミンＣを加えることが好ましく、炭酸カルシウム、及びビタミンＣを加えることがより好ましい。 -Other additives-
The other additives are appropriately selected depending on the intended purpose without any limitation, and examples thereof include calcium carbonate, vitamin C, citric acid, NaCl solution, and distilled water.
These other additives may be used alone or in combination of two or more, but they promote acceleration of neutralization of lactic acid which is a metabolite of lactic acid bacteria, anaerobic fermentation efficiency, and Bacillus natto. From the viewpoint of suppressing the growth of the above, it is preferable to add calcium carbonate or vitamin C, and it is more preferable to add calcium carbonate and vitamin C.

前記炭酸カルシウムの前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、炭酸カルシウム０．１〜２０質量部が好ましく、０．１〜１０質量部がより好ましく、０．５〜５質量部が更に好ましい。
前記ビタミンＣの前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、ビタミンＣ０．１〜１０質量部が好ましく、０．１〜５質量部がより好ましく、０．５〜３質量部が更に好ましい。
前記クエン酸の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、クエン酸０．１〜１０質量部が好ましく、０．１〜５質量部がより好ましく、０．５〜３質量部が更に好ましい。 The amount of the calcium carbonate with respect to the natto is not particularly limited and may be appropriately selected depending on the purpose, but is preferably 0.1 to 20 parts by mass of calcium carbonate with respect to 100 parts by mass of natto, and 0. 1 to 10 parts by mass is more preferable, and 0.5 to 5 parts by mass is further preferable.
The amount of the vitamin C with respect to the natto is not particularly limited and can be appropriately selected depending on the purpose, but 0.1 to 10 parts by mass of vitamin C is preferable with respect to 100 parts by mass of natto, 0.1 ˜5 parts by mass is more preferred, and 0.5 to 3 parts by mass is even more preferred.
The amount of the citric acid with respect to the natto is not particularly limited and may be appropriately selected depending on the purpose, but is preferably 0.1 to 10 parts by mass of citric acid with respect to 100 parts by mass of natto, and 0.1. 1 to 5 parts by mass is more preferable, and 0.5 to 3 parts by mass is further preferable.

前記ＮａＣｌ溶液の濃度としては、目的に応じて適宜選択することができるが、０．０１〜１０％溶液が好ましく、０．１〜１．０％溶液がより好ましく、０．５〜１．０％溶液が更に好ましく、０．９％溶液が特に好ましい。
前記ＮａＣｌ溶液の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、ＮａＣｌ溶液０．１〜２０質量部が好ましく、１．０〜１０質量部がより好ましく、３．０〜１０質量部が更に好ましい。
前記蒸留水の前記納豆に対する量としては、特に制限はなく、目的に応じて適宜選択することができるが、納豆１００質量部に対して、蒸留水１〜１００質量部が好ましく、１０〜１００質量部がより好ましく、２０〜４０質量部が更に好ましい。 The concentration of the NaCl solution can be appropriately selected depending on the purpose, but is preferably 0.01 to 10% solution, more preferably 0.1 to 1.0% solution, and 0.5 to 1.0. % Solutions are more preferred, and 0.9% solutions are especially preferred.
The amount of the NaCl solution with respect to the natto is not particularly limited and may be appropriately selected depending on the purpose, but is preferably 0.1 to 20 parts by mass of the NaCl solution with respect to 100 parts by mass of natto. 0 to 10 parts by mass is more preferable, and 3.0 to 10 parts by mass is further preferable.
The amount of the distilled water with respect to the natto is not particularly limited and can be appropriately selected depending on the purpose, but 1 to 100 parts by mass of distilled water is preferable with respect to 100 parts by mass of natto, and 10 to 100 parts by mass. A part is more preferable, and 20-40 mass parts is still more preferable.

＜その他の工程＞
前記食品の製造方法における、前記その他の工程としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、真空凍結乾燥工程、通風乾燥工程などが挙げられる。 <Other processes>
The other steps in the method for producing a food are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a vacuum freeze drying step and a ventilation drying step.

＜食品＞
以上の工程により本発明の食品の製造方法により製造される食品を得ることができる。
前記食品は、少なくとも、納豆菌と、乳酸菌、酪酸菌、及び麹菌の少なくともいずれかとを含み、更に必要に応じて、その他の成分を含むことができる。 <Food>
Through the above steps, a food product manufactured by the food manufacturing method of the present invention can be obtained.
The food contains at least Bacillus natto and at least one of lactic acid bacterium, butyric acid bacterium, and koji mold, and may further contain other components as necessary.

前記食品における、納豆菌、乳酸菌、酪酸菌、及び麹菌の濃度としては、特に制限はなく、目的に応じて適宜選択することができるが、前記納豆菌、乳酸菌、酪酸菌、及び麹菌の濃度の少なくともいずれかが、生菌で、１×１０^８個／ｇ以上であることが好ましく、前記納豆菌、乳酸菌、酪酸菌、及び麹菌の濃度の少なくともいずれかが、生菌で、１×１０^９個／ｇ以上であることがより好ましく、納豆菌の濃度が、生菌で、１×１０^８個／ｇ以上であり、かつ、乳酸菌、酪酸菌、及び麹菌の濃度の少なくともいずれかが、生菌で、１×１０^８個／ｇ以上であることがさらに好ましく、納豆菌の濃度が、生菌で、１×１０^９個／ｇ以上であり、かつ、乳酸菌、酪酸菌、及び麹菌の濃度の少なくともいずれかが、生菌で、１×１０^９個／ｇ以上であることが特に好ましい。 In the food, the concentration of Bacillus natto, lactic acid bacterium, butyric acid bacterium, and koji mold are not particularly limited and can be appropriately selected according to the purpose, but the concentration of natto bacterium, lactic acid bacterium, butyric acid bacterium, and koji mold It is preferable that at least one of the live bacteria is 1×10 ⁸ cells/g or more, and at least one of the concentrations of the natto bacterium, the lactic acid bacterium, the butyric acid bacterium, and the koji mold is the live bacterium, 1×10 ⁹ More preferably, the concentration of natto bacteria is 1×10 ⁸ cells/g or more, and at least one of the concentrations of lactic acid bacteria, butyric acid bacteria, and koji mold is raw. More preferably, the concentration of natto is 1×10 ⁸ cells/g or more, and the concentration of natto is 1×10 ⁹ cells/g or more, and the concentration of lactic acid bacteria, butyric acid bacteria, and koji mold. It is particularly preferable that at least any one of them is a viable bacterium, and is 1×10 ⁹ cells/g or more.

前記食品におけるプロテアーゼ力価としては、特に制限はなく、目的に応じて適宜選択することができるが、１，０００ｕ／ｇ以上であることが好ましく、５，０００ｕ／ｇ以上であることがより好ましく、８，０００ｕ／ｇ以上であることが更に好ましい。 The protease titer in the food is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 1,000 u/g or more, more preferably 5,000 u/g or more. , 8,000 u/g or more is more preferable.

前記食品におけるプロテアーゼ力価は、５ｇの試料に、２５ｍＬの０．５％ＮａＣｌ溶液を加え撹拌し、遠心後の上清０．５ｍＬに、１．０ｍＬのマッキンベン緩衝液（ｐＨ６）を加えて、３８℃で５分間予熱し、１．５ｍＬの２％カゼイン溶液を加えて、３８℃で６０分間保温した後、３．０ｍＬの０．４Ｍトリクロロ酢酸を加えて、３０分後にろ過し、１．０ｍＬのろ液に、５．０ｍＬの０．４Ｍ炭酸ナトリウム溶液、１．０ｍＬのフェノール試薬を加えて、３８℃で３０分間発色させ、６６０ｎｍにおける吸光度を測定し、チロシン標準液を用いて作成した検量線から、３８℃、６０分間に1μｇのチロシンを生成する活性を1単位として算出することができる。 The protease titer in the food was obtained by adding 25 mL of 0.5% NaCl solution to 5 g of the sample and stirring, and adding 1.0 mL of McKinben's buffer solution (pH 6) to 0.5 mL of the supernatant after centrifugation, Preheat at 38° C. for 5 minutes, add 1.5 mL of 2% casein solution, keep warm at 38° C. for 60 minutes, add 3.0 mL of 0.4 M trichloroacetic acid, and filter after 30 minutes, 1. 5.0 mL of 0.4 M sodium carbonate solution and 1.0 mL of phenol reagent were added to 0 mL of the filtrate, color was developed at 38° C. for 30 minutes, the absorbance at 660 nm was measured, and the solution was prepared using tyrosine standard solution. From the calibration curve, the activity of producing 1 μg of tyrosine at 38° C. for 60 minutes can be calculated as 1 unit.

以下、本発明の実施例を説明するが、本発明は、これらの実施例に何ら限定されるものではない。 Examples of the present invention will be described below, but the present invention is not limited to these examples.

＜納豆の乳酸菌発酵による食品の製造＞
（実施例１−１）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。３０ｇの粗挽きした納豆と、３００ｍｇの有胞子乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ（三菱化学フーズ製）とを混合し、３５℃で好気発酵した。発酵後４８時間の乳酸菌の生菌数を、以下の方法で測定し、結果を図１に示した。 <Production of food by fermenting natto lactic acid bacteria>
(Example 1-1)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 30 g of coarsely ground natto was mixed with 300 mg of Lactobacillus sporogenas (manufactured by Mitsubishi Kagaku Foods), a spore lactic acid bacterium, and aerobically fermented at 35°C. The viable cell count of lactic acid bacteria 48 hours after fermentation was measured by the following method, and the results are shown in FIG.

−好気発酵乳酸菌の生菌数の測定−
５ｇの試料に、１００ｍＬの滅菌済み０．９％ＮａＣｌ溶液を加え、室温で３０分間撹拌し、乳酸菌を抽出した。抽出物を、滅菌済み０．９％ＮａＣｌ溶液を用いて希釈し、ロゴサ寒天培地(関東化学株式会社)を氷酢酸を加えずに調整した（ｐＨ５）プレート半面に５０μＬをコンラージした。３５℃で４８時間培養した後に、菌数を測定した。本条件下では納豆菌は繁殖しないため、全てのコロニーを乳酸菌として計測した。 -Measurement of viable cell count of aerobic fermenting lactic acid bacteria-
To 5 g of the sample, 100 mL of sterilized 0.9% NaCl solution was added and stirred at room temperature for 30 minutes to extract lactic acid bacteria. The extract was diluted with a sterilized 0.9% NaCl solution, and Rogosa agar medium (Kanto Kagaku Co., Ltd.) was adjusted without adding glacial acetic acid (pH 5), and 50 μL was confluent on one half of the plate. After culturing at 35° C. for 48 hours, the number of bacteria was measured. Since Bacillus natto does not reproduce under these conditions, all colonies were counted as lactic acid bacteria.

（実施例１−２）
粗挽きした納豆と、有胞子乳酸菌に加えて、１．５ｇの炭酸カルシウム（関東化学株式会社製）及び０．３ｇのビタミンＣ（中国江蘇江山製薬製）を添加した点以外は、実施例１−１と同様の発酵及び測定を行った。結果を図１に示した。 (Example 1-2)
Example 1 except that 1.5 g of calcium carbonate (manufactured by Kanto Chemical Co., Inc.) and 0.3 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical Co., Ltd.) were added to the coarsely ground natto and spore lactic acid bacteria. The same fermentation and measurement as in -1 were performed. The results are shown in Fig. 1.

（比較例１−１）
２００ｍｌ広口コルベンに６時間浸漬した８０ｇの大豆を入れて、３０分蒸煮した。冷却後に、蒸煮した大豆と、８００ｍｇの有胞子乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ（三菱化学フーズ製）とを混合し、３５℃で好気発酵した。発酵後４８時間の乳酸菌の生菌数を実施例１−１と同様の方法で測定し、結果を図１に示した。 (Comparative Example 1-1)
80 g of soybeans soaked in 200 ml wide-mouth Kolben for 6 hours were put and steamed for 30 minutes. After cooling, the steamed soybeans were mixed with 800 mg of Lactobacillus sporogenes (manufactured by Mitsubishi Kagaku Foods), and aerobically fermented at 35°C. The viable cell count of lactic acid bacteria 48 hours after fermentation was measured by the same method as in Example 1-1, and the results are shown in FIG.

（比較例１−２）
蒸煮した大豆と、有胞子乳酸菌に加えて、２．４ｇの炭酸カルシウム（関東化学株式会社製）及び０．８ｇのビタミンＣ（中国江蘇江山製薬製）を添加した点以外は、比較例１−１と同様の発酵及び測定を行った。結果を図１に示した。 (Comparative Example 1-2)
Comparative Example 1-except that 2.4 g of calcium carbonate (manufactured by Kanto Chemical Co., Inc.) and 0.8 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical Co., Ltd.) were added to the steamed soybean and the spore-forming lactic acid bacterium. The same fermentation and measurement as in 1 were performed. The results are shown in Fig. 1.

（実施例２−１）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。４０ｇの粗挽きした納豆と、１２０ｍｇの有胞子乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ（三菱化学フーズ製）とを混合し、０．４ｇのビタミンＣ（中国江蘇江山製薬製）、２ｇの炭酸カルシウム（関東化学株式会社製）、及び蒸留水１２ｍＬを加え、３５℃で好気発酵した。発酵前、発酵後２４時間、及び発酵後４８時間の乳酸菌の生菌数を、実施例１−１と同様の方法で測定し、結果を図２に示した（マーカーが●の折れ線）。発酵前、発酵後２４時間、及び発酵後４８時間の納豆菌の菌数を以下の方法で測定し、結果を図３に示した（マーカーが●の折れ線）。 (Example 2-1)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 40 g of coarsely ground natto and 120 mg of Lactobacillus sporogenes (manufactured by Mitsubishi Kagaku Foods), a spore lactic acid bacterium, are mixed, and 0.4 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical, China) and 2 g of calcium carbonate (manufactured by Kanto Kagaku Co. ), and 12 mL of distilled water were added, and aerobically fermented at 35°C. The viable cell counts of lactic acid bacteria before fermentation, 24 hours after fermentation, and 48 hours after fermentation were measured by the same method as in Example 1-1, and the results are shown in FIG. 2 (marker is a polygonal line). The number of Bacillus natto before and after the fermentation, 24 hours after the fermentation, and 48 hours after the fermentation was measured by the following method, and the results are shown in FIG. 3 (marker is a polygonal line).

−納豆菌の生菌数の測定−
５ｇの試料に、１００ｍＬの滅菌済み０．９％ＮａＣｌ溶液を加え、室温で３０分間撹拌し、納豆菌を抽出した。抽出物を、滅菌済み０．９％ＮａＣｌ溶液を用いて希釈し、シカメディアＢＣＰ加プレートカウント寒天培地(関東化学株式会社)で調製したプレート半面に５０μＬをコンラージした。３５℃で４８時間培養した後に、菌数を測定し、結果を図１に示した。小さい点のようなコロニーを乳酸菌として除外し、大きく広がるコロニーを納豆菌として計測した。 -Measurement of viable count of Bacillus natto-
100 mL of sterilized 0.9% NaCl solution was added to 5 g of the sample, and the mixture was stirred at room temperature for 30 minutes to extract Bacillus natto. The extract was diluted with a sterilized 0.9% NaCl solution, and 50 μL of the plate was prepared on a plate count agar medium (Kanto Kagaku Co., Ltd.) containing Sikamedia BCP (50 μL). After culturing at 35° C. for 48 hours, the number of bacteria was measured, and the results are shown in FIG. Colonies such as small dots were excluded as lactic acid bacteria, and large spread colonies were counted as Bacillus natto.

（実施例２−２）
炭酸カルシウムを添加しない点以外は、実施例２−１と同様の発酵及び測定を行った。結果を図２及び図３に示した（マーカーが▲の折れ線）。 (Example 2-2)
Fermentation and measurement were performed in the same manner as in Example 2-1, except that calcium carbonate was not added. The results are shown in FIG. 2 and FIG. 3 (marker is lined with ▲).

（実施例２−３）
炭酸カルシウム及びビタミンＣに代えて、０．４ｇのクエン酸（クエン酸一水和物、和光純薬工業株式会社製）を添加した以外は、実施例２−１と同様の発酵及び測定を行った。結果を図２及び図３に示した（マーカーが■の折れ線）。 (Example 2-3)
Fermentation and measurement were performed in the same manner as in Example 2-1, except that 0.4 g of citric acid (citric acid monohydrate, manufactured by Wako Pure Chemical Industries, Ltd.) was added instead of calcium carbonate and vitamin C. It was The results are shown in FIG. 2 and FIG. 3 (marker is a polygonal line).

（比較例２）
乳酸菌を加えない点以外は、実施例２−３と同様の発酵及び測定を行った。結果を図２及び図３に示した（マーカーなしの折れ線）。 (Comparative example 2)
Fermentation and measurement were performed in the same manner as in Example 2-3, except that lactic acid bacteria were not added. The results are shown in FIGS. 2 and 3 (the broken line without markers).

＜納豆の酪酸菌発酵＞
（実施例３）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。５００ｇの粗挽きした納豆と、２．５ｇの酪酸菌Ｃｌｏｓｔｒｉｄｉｕｍｂｕｔｙｒｉｃｕｍ（東亜薬品工業製）とを混合し、１０ｇの炭酸カルシウム（関東化学株式会社製）、及び５０ｍＬの０．９％ＮａＣｌを加え、アネロパック・ケンキ（三菱ガス化学製）、及び角型ジャー（三菱ガス化学製）を用いて３５℃で嫌気発酵した。発酵前、発酵後２４時間、発酵後４８時間、発酵後７２時間、及び発酵後９６時間の酪酸菌の生菌数を、以下の方法で測定し、結果を図４に示した（マーカーが▲の折れ線）。発酵前、発酵後２４時間、及び発酵後４８時間の納豆菌の生菌数を実施例２−１と同様の方法で測定し、結果を図４に示した（マーカーが●の折れ線）。 <Fermentation of natto butyric acid bacteria>
(Example 3)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 500 g of coarsely ground natto and 2.5 g of butyric acid bacterium Clostridium butyricum (manufactured by Toa Pharmaceutical Co., Ltd.) were mixed, 10 g of calcium carbonate (manufactured by Kanto Chemical Co., Inc.), and 50 mL of 0.9% NaCl were added, Anaerobic fermentation was performed at 35° C. using Aneropack Kenki (manufactured by Mitsubishi Gas Chemical Co., Inc.) and a rectangular jar (manufactured by Mitsubishi Gas Chemical Co., Ltd.). The viable cell count of butyric acid bacteria before fermentation, 24 hours after fermentation, 48 hours after fermentation, 72 hours after fermentation, and 96 hours after fermentation was measured by the following method, and the results are shown in FIG. Line). The number of viable Bacillus natto before and after fermentation, and after 24 hours and 48 hours after fermentation was measured by the same method as in Example 2-1, and the results are shown in FIG. 4 (marker is a polygonal line).

−酪酸菌の生菌数の測定−
５ｇの試料に、１００ｍＬの滅菌済み０．９％ＮａＣｌ溶液を加え、室温で３０分間撹拌し、酪酸菌を抽出した。抽出物を、滅菌済み０．９％ＮａＣｌ溶液を用いて希釈し、標準寒天培地(日水製薬株式会社)をチオグリコール酸ナトリウムおよびL-システイン塩酸塩一水和物を０．３％加えて調整したプレート半面に５０μＬをコンラージした。アネロパック・ケンキ（三菱ガス化学製）、及び角型ジャー（三菱ガス化学製）を用いて４８時間３５℃で嫌気培養した後に、菌数を測定し、結果を図１に示した。なお、本条件下では、納豆菌は繁殖しないのですべてのコロニーを酪酸菌として計測した。 -Measurement of the viable cell count of butyric acid bacteria-
100 mL of sterilized 0.9% NaCl solution was added to 5 g of the sample, and the mixture was stirred at room temperature for 30 minutes to extract butyric acid bacteria. The extract was diluted with a sterilized 0.9% NaCl solution, and standard agar medium (Nissui Pharmaceutical Co., Ltd.) was added with sodium thioglycolate and 0.3% of L-cysteine hydrochloride monohydrate. 50 μL of the prepared plate half surface was confluent. Anaerobic culture was performed for 48 hours at 35° C. using Aneropack Kenki (manufactured by Mitsubishi Gas Chemical Co., Inc.) and a rectangular jar (manufactured by Mitsubishi Gas Chemical Co., Inc.), and the number of bacteria was measured. The results are shown in FIG. Under these conditions, Bacillus natto does not propagate, so all colonies were counted as butyric acid bacteria.

（実施例４）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。５８０ｇの粗挽きした納豆と、２．９ｇの酪酸菌Ｃｌｏｓｔｒｉｄｉｕｍｂｕｔｙｒｉｃｕｍ（東亜薬品工業製）とを混合し、１０ｇの炭酸カルシウム（関東化学株式会社製）、及び５０ｍＬの０．９％ＮａＣｌを加え、アネロパック・ケンキ（三菱ガス化学製）、及び角型ジャー（三菱ガス化学製）を用いて３５℃で４８時間嫌気発酵し、得られた発酵物を、凍結保存した後、真空凍結乾燥及び通風乾燥した。乾燥前、及び乾燥後の酪酸菌の生菌数を、実施例３と同様の方法で測定し、結果を図５に示した。発酵乾燥物のプロテアーゼ力価を、以下の方法で測定したところ、乾燥粉末あたり８，８２９ｕ／ｇであり、原料あたり３，３２７ｕ／ｇであった。 (Example 4)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 580 g of coarsely ground natto and 2.9 g of butyric acid bacterium Clostridium butyricum (manufactured by Toa Pharmaceutical Co., Ltd.) were mixed, 10 g of calcium carbonate (manufactured by Kanto Kagaku Co., Ltd.), and 50 mL of 0.9% NaCl were added, Anaerobic Kenki (manufactured by Mitsubishi Gas Chemical) and square jar (manufactured by Mitsubishi Gas Chemical) were anaerobically fermented at 35°C for 48 hours, and the resulting fermented product was frozen and stored, and then vacuum freeze-dried and ventilated. did. The viable cell counts of butyric acid bacteria before and after drying were measured by the same method as in Example 3, and the results are shown in FIG. When the protease titer of the fermented dried product was measured by the following method, it was 8,829 u/g per dry powder and 3,327 u/g per raw material.

−プロテアーゼ力価の測定−
５ｇの試料に、２５ｍＬの０．５％ＮａＣｌ溶液を加え、室温で３時間撹拌し、３，０００ｒｐｍで１５分間遠心した。遠心後の上清０．５ｍＬに、１．０ｍＬのマッキンベン緩衝液（ｐＨ６）を加えて、３８℃で５分間予熱した。１．５ｍＬのカゼイン溶液を加えて、３８℃で６０分間保温した。３．０ｍＬの０．４Ｍトリクロロ酢酸を加えて、３０分後にろ過した。１．０ｍＬのろ液に、５．０ｍＬの０．４Ｍ炭酸ナトリウム溶液、１．０ｍＬのフェノール試薬（関東化学株式会社製）を加えて、３８℃で３０分間発色させた。６６０ｎｍにおける吸光度を測定し、２５、５０、７５μｇ／ｍＬのチロシン標準液を用いて作成した検量線から、３８℃、６０分間に1μｇのチロシンを生成する活性を1単位としてプロテアーゼ力価を算出した。 -Measurement of protease titer-
To 5 g of the sample, 25 mL of 0.5% NaCl solution was added, stirred at room temperature for 3 hours, and then centrifuged at 3,000 rpm for 15 minutes. To 0.5 mL of the supernatant after centrifugation, 1.0 mL of McKinben's buffer (pH 6) was added and preheated at 38° C. for 5 minutes. 1.5 mL of casein solution was added, and the mixture was kept at 38° C. for 60 minutes. 3.0 mL of 0.4 M trichloroacetic acid was added, and after 30 minutes, filtration was performed. To 1.0 mL of the filtrate, 5.0 mL of 0.4 M sodium carbonate solution and 1.0 mL of phenol reagent (manufactured by Kanto Kagaku Co., Ltd.) were added, and color was developed at 38° C. for 30 minutes. The absorbance at 660 nm was measured, and the protease titer was calculated from the calibration curve prepared using 25, 50, and 75 μg/mL tyrosine standard solution, with the activity of producing 1 μg tyrosine at 38° C. and 60 minutes as 1 unit. ..

−−マッキンベン緩衝液（ｐＨ６）−−
７１．６３ｇのリン酸水素二ナトリウム（Ｎａ_２ＨＰＯ_４・１２Ｈ_２Ｏ）を蒸留水に溶解し、１，０００ｍＬに定容し、Ａ液を作成した。
２１ｇのクエン酸（Ｃ_６Ｈ_８Ｏ_７・Ｈ_２Ｏ）蒸留水に溶解し、１，０００ｍＬに定容し、Ｂ液を作成した。
Ａ液とＢ液とをｐＨメーター（ｐＨｓｉｏｎＣ−７３アズワン株式会社製）を用いて、ｐＨ６．０になるように混合した。 ---McKimben buffer (pH 6)---
Disodium hydrogen phosphate 71.63g of _{_{_{(Na 2 HPO 4 · 12H 2}}} O) was dissolved in distilled water, being diluted to 1,000 mL, was created A solution.
21 g of citric acid (C ₆ H ₈ O ₇ .H ₂ O) was dissolved in distilled water and the volume was adjusted to 1,000 mL to prepare solution B.
The liquid A and the liquid B were mixed using a pH meter (pHsion C-73 As One Co., Ltd.) so as to have a pH of 6.0.

−−カゼイン溶液−−
１００ｍＬビーカーに、２ｇのカゼイン、及び少量の蒸留水を加え、ガラス棒でよく混濁後、１３ｍＬの０．１Ｍ水酸化ナトリウムを加え徐々に加熱し、沸騰溶解した。更に、沸騰加熱しながら２ｍＬの０．５Ｍ乳酸を一滴ずつ滴下し、十分に溶解させた。水で急冷させ、０．２ＭＮａ_２ＨＰＯ_４を用いてｐＨ６．０に調整し、２０ｍＬのマッキンベン緩衝液（ｐＨ６）を加え、１００ｍＬに定容した。 --- Casein solution ---
To a 100 mL beaker, 2 g of casein and a small amount of distilled water were added, and after thoroughly clouding with a glass rod, 13 mL of 0.1 M sodium hydroxide was added and gradually heated to dissolve by boiling. Furthermore, 2 mL of 0.5 M lactic acid was added dropwise drop by drop while heating with boiling to sufficiently dissolve it. It was quenched with water, adjusted to pH 6.0 with 0.2 M Na ₂ HPO ₄ , 20 mL of McKinben's buffer (pH 6) was added, and the volume was adjusted to 100 mL.

＜納豆の麹菌発酵＞
（実施例５）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。２００ｇの粗挽きした納豆と、３００ｍｇの清酒用種麹（株式会社ビオック製）とを混合し、３５℃で好気発酵した。発酵後９６時間の発酵物の写真を図６に示した。図６に示したとおり、白色の麹菌が観察された。発酵物の顕微鏡写真を図７に示した。図７に示したとおり、糸状の麹菌が観察された。発酵後９６時間の納豆菌の生菌数を、実施例２−１と同様の方法で測定したところ、３．７×１０^９個／ｇであった。発酵後９６時間のプロテアーゼ力価を、実施例４と同様の方法で測定したところ、乾燥粉末あたり１１，０００ｕ／ｇであった。 <Koji fermentation of natto>
(Example 5)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 200 g of coarsely ground natto and 300 mg of rice malt for sake (manufactured by Bioc Co., Ltd.) were mixed and aerobically fermented at 35°C. A photograph of the fermented product 96 hours after fermentation is shown in FIG. As shown in FIG. 6, white koji mold was observed. A micrograph of the fermented product is shown in FIG. As shown in FIG. 7, filamentous koji mold was observed. The viable cell count of Bacillus natto after 96 hours of fermentation was measured by the same method as in Example 2-1, and it was 3.7×10 ⁹ cells/g. When the protease titer at 96 hours after fermentation was measured by the same method as in Example 4, it was 11,000 u/g per dry powder.

＜納豆の少なくとも２種類の乳酸菌による発酵＞
（実施例６−１）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。６００ｇの粗挽きした納豆と、６ｇの有胞子乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ（三菱化学フーズ製）とを混合し、６ｇのビタミンＣ（中国江蘇江山製薬製）、３０ｇの炭酸カルシウム（関東化学株式会社製）、及び１８０ｍＬの蒸留水を加え、３５℃で好気発酵した。発酵前、発酵後７２時間、及び発酵後１２０時間の乳酸菌の生菌数を、実施例１−１と同様に測定し、結果を図８に示した（マーカーが●の折れ線）。発酵前、発酵後７２時間、及び発酵後１２０時間の納豆菌の菌数を実施例２−１と測定し、結果を図９に示した（マーカーが●の折れ線）。発酵乾燥物のプロテアーゼ力価を、実施例４と同様の方法で測定したところ、乾燥粉末あたり９，２００ｕ／ｇであった。 <Fermentation of natto with at least two types of lactic acid bacteria>
(Example 6-1)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 600 g of coarsely ground natto and 6 g of spore lactic acid bacterium Lactobacillus sporogenes (manufactured by Mitsubishi Kagaku Foods) were mixed, and 6 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical Co., Ltd. in China), 30 g of calcium carbonate (manufactured by Kanto Kagaku Co., Ltd.), And 180 mL of distilled water were added, and aerobically fermented at 35°C. The viable cell counts of lactic acid bacteria before fermentation, 72 hours after fermentation, and 120 hours after fermentation were measured in the same manner as in Example 1-1, and the results are shown in FIG. 8 (marker is a polygonal line). The number of Bacillus subtilis natto before fermentation, 72 hours after fermentation, and 120 hours after fermentation was measured as in Example 2-1, and the results are shown in FIG. 9 (marker is a polygonal line). The protease titer of the fermented dried product was measured by the same method as in Example 4, and it was 9,200 u/g per dry powder.

（実施例６−２）
乳酸菌を、合計量が６ｇの、以下の乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓａｃｉｄｏｐｈｉｌｕｓ、Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍｌｏｎｇｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｃａｓｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｐｌａｎｔｒｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓａｋｅｉ、及びＬａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ：有限会社ラヴィアンサンテ製）に代えた以外は、実施例６−１と同様の発酵及び測定を行った。結果を図８及び図９に示した（マーカーが▲の折れ線）。発酵乾燥物のプロテアーゼ力価を、実施例４と同様の方法で測定したところ、乾燥粉末あたり８，４００ｕ／ｇであった。 (Example 6-2)
The total amount of lactic acid bacteria is 6 g, and the total amount of the lactic acid bacteria is Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, Lactobacillus plant sprout, and Lactobacillus sp. The same fermentation and measurement were performed. The results are shown in FIG. 8 and FIG. 9 (marker is lined with ▲). When the protease titer of the dried fermentation product was measured by the same method as in Example 4, it was 8,400 u/g per dry powder.

（実施例７−１）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。６２０ｇの粗挽きした納豆と、合計量が６．２ｇの乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓａｃｉｄｏｐｈｉｌｕｓ、Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍｌｏｎｇｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｃａｓｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｐｌａｎｔｒｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓａｋｅｉ、及びＬａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ：有限会社ラヴィアンサンテ製）とを混合し、３０ｍＬの０．９％ＮａＣｌを加え、３５℃で好気発酵した。発酵後４８時間の乳酸菌の生菌数を、実施例１−１と同様に測定し、結果を図１０に示した（１）。発酵後４８時間の納豆菌の菌数を実施例２−１と同様の方法で測定し、結果を図１０に示した（２）。 (Example 7-1)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. And natto was coarse ground of 620g, the total amount of 6.2g lactic acid bacteria (Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, Lactobacillus plantrum, Lactobacillus sakei, and Lactobacillus sporogenas: limited company made Ravi Anne Sante) were mixed, 0 of 30mL 0.9% NaCl was added, and aerobically fermented at 35°C. The viable cell count of lactic acid bacteria 48 hours after fermentation was measured in the same manner as in Example 1-1, and the results are shown in FIG. 10 (1). The number of Bacillus natto bacteria 48 hours after fermentation was measured by the same method as in Example 2-1, and the results are shown in FIG. 10 (2).

（実施例７−２）
発酵工程において、５ｇのビタミンＣ（中国江蘇江山製薬製）を加えた以外は、実施例７−１と同様の発酵及び測定を行った。結果を図１０に示した。 (Example 7-2)
In the fermentation process, the same fermentation and measurement as in Example 7-1 were performed except that 5 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical Co., Ltd. of China) was added. The results are shown in Fig. 10.

＜納豆の乳酸菌による嫌気発酵＞
（実施例８−１）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。６２０ｇの粗挽きした納豆と、合計量が６．２ｇの乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓａｃｉｄｏｐｈｉｌｕｓ、Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍｌｏｎｇｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｃａｓｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｐｌａｎｔｒｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓａｋｅｉ、及びＬａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ：有限会社ラヴィアンサンテ製）とを混合し、６ｇの炭酸カルシウム（関東化学株式会社製）、及び３０ｍＬの０．９％ＮａＣｌを加え、３５℃で嫌気パック（アネロパック・ケンキ、三菱ガス化学株式会社製）、及び嫌気ジャー（角型ジャー、三菱ガス化学株式会社製）を用いて嫌気発酵した。発酵前、発酵後２４時間、発酵後４８時間、発酵後７２時間、及び９６時間の乳酸菌の生菌数を以下の方法で測定し、結果を図１１に示した（マーカーが▲の点線）。発酵物のｐＨ値を以下の方法で測定し、結果を図１１に示した（マーカーが▲の折れ線）。 <Anaerobic fermentation of natto with lactic acid bacteria>
(Example 8-1)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 620 g of coarsely ground natto, and a total amount of 6.2 g of lactic acid bacteria (Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, Lactobacillus planta, Lactobacillus planta, Lactobacillus planta, Lactobacillus lactobacillus, and Lactobacillus lactobacillus sp. Calcium (manufactured by Kanto Chemical Co., Inc.) and 30 mL of 0.9% NaCl were added, and an anaerobic pack (Aneropack Kenki, manufactured by Mitsubishi Gas Chemical Co., Inc.) and an anaerobic jar (square jar, Mitsubishi Gas Chemical Co., Ltd.) were added at 35°C. Anaerobic fermentation was carried out using a product manufactured by the company. The viable cell counts of lactic acid bacteria before fermentation, 24 hours after fermentation, 48 hours after fermentation, 72 hours after fermentation, and 96 hours after fermentation were measured by the following method, and the results are shown in Fig. 11 (marked with a dotted line). The pH value of the fermented product was measured by the following method, and the results are shown in FIG. 11 (marked with a broken line).

−嫌気発酵菌の生菌数の測定−
５ｇの試料に、１００ｍＬの滅菌済み０．９％ＮａＣｌ溶液を加え、室温で３０分間撹拌し、乳酸菌を抽出した。抽出物を、滅菌済み０．９％ＮａＣｌ溶液を用いて希釈し、ロゴサ寒天培地(関東化学株式会社製)に、チオグリコール酸ナトリウム（関東化学株式会社製）及びＬ−システイン塩酸塩一水和物（関東化学株式会社製）を０．３％加え、氷酢酸を加えずに調整したプレート半面に５０μＬをコンラージした。３５℃で嫌気パック（アネロパック・ケンキ、三菱ガス化学株式会社製）、及び嫌気ジャー（角型ジャー、三菱ガス化学株式会社製）を用いて４８時間３５℃で嫌気発酵した後に、菌数を測定した。なお、本条件下では納豆菌は繁殖しないため、全てのコロニーを乳酸菌として計測した。 -Measurement of the viable cell count of anaerobic fermenting bacteria-
To 5 g of the sample, 100 mL of sterilized 0.9% NaCl solution was added and stirred at room temperature for 30 minutes to extract lactic acid bacteria. The extract was diluted with a sterilized 0.9% NaCl solution, and was added to Rogosa agar medium (Kanto Chemical Co., Inc.), sodium thioglycolate (Kanto Chemical Co., Ltd.) and L-cysteine hydrochloride monohydrate. (Kanto Kagaku Co., Ltd.) was added in an amount of 0.3%, and 50 μL of the plate was adjusted to the half surface of the plate prepared without adding glacial acetic acid. After anaerobic fermentation at 35°C for 48 hours using an anaerobic pack (Aneropack Kenki, Mitsubishi Gas Chemical Co., Ltd.) and an anaerobic jar (square jar, Mitsubishi Gas Chemical Co., Ltd.) at 35°C, the number of bacteria is measured. did. In addition, since Bacillus natto does not reproduce under these conditions, all colonies were counted as lactic acid bacteria.

−ｐＨ値測定−
５ｇの試料に、２５ｍＬの０．５％ＮａＣｌ溶液を加え、ｐＨメーター（ｐＨｓｉｏｎＣ−７３アズワン株式会社製）を用いてｐＨ値を測定した。 -PH value measurement-
To 5 g of the sample, 25 mL of 0.5% NaCl solution was added, and the pH value was measured using a pH meter (pHsion C-73 As One Co., Ltd.).

（実施例８−２）
炭酸カルシウムを加えない以外は、実施例８−１と同様の発酵及び測定を行い、乳酸菌及び酪酸菌の生菌数を図１１に示した（マーカーが●の点線）。発酵物のｐＨ値の結果を図１０に示した（マーカーが●の折れ線）。 (Example 8-2)
Fermentation and measurement were carried out in the same manner as in Example 8-1, except that calcium carbonate was not added, and the viable cell counts of lactic acid bacteria and butyric acid bacteria are shown in FIG. 11 (marker indicates dotted line). The result of pH value of the fermented product is shown in FIG. 10 (marker is a polygonal line).

（実施例９）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。３００ｇの粗挽きした納豆と、合計量が４．３ｇの乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓａｃｉｄｏｐｈｉｌｕｓ、Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍｌｏｎｇｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｃａｓｅｉ、Ｌａｃｔｏｂａｃｉｌｌｕｓｐｌａｎｔｒｕｍ、Ｌａｃｔｏｂａｃｉｌｌｕｓｓａｋｅｉ、及びＬａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ：有限会社ラヴィアンサンテ製）とを混合し、６ｇの炭酸カルシウム（関東化学株式会社製）、及び１５ｍＬの０．９％ＮａＣｌを加え、３５℃で嫌気パック（アネロパック・ケンキ、三菱ガス化学株式会社製）、及び嫌気ジャー（角型ジャー、三菱ガス化学株式会社製）を用いて嫌気発酵した。発酵前、発酵後２４時間、発酵後４８時間、発酵後７２時間、発酵後１４４時間の乳酸菌の生菌数を実施例８８−１と同様に測定し、結果を図１２に示した（マーカーが●の折れ線）。発酵物の納豆のプロテアーゼ力価を実施例４と同様の方法で測定し、結果を図１２に示した（マーカーが■の折れ線）。 (Example 9)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. Natto was coarse-ground in 300 g, the total amount of 4.3g of lactic acid bacteria (Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, Lactobacillus plantrum, Lactobacillus sakei, and Lactobacillus sporogenas: Co. Ravi en Ltd. Sante) were mixed, 6 g of carbonate Calcium (Kanto Chemical Co., Inc.) and 15 mL of 0.9% NaCl were added, and anaerobic pack (Aneropack Kenki, Mitsubishi Gas Chemical Co., Inc.) and anaerobic jar (square jar, Mitsubishi Gas Chemical stock) were added at 35°C. Anaerobic fermentation was carried out using a product manufactured by the company. Pre-fermentation, post-fermentation 24 hours, post-fermentation 48 hours, post-fermentation 72 hours, post-fermentation 144 hours post-fermentation lactic acid bacteria viable count was measured in the same manner as in Example 88-1, and the results are shown in FIG. ● Line). The protease titer of natto of the fermented product was measured by the same method as in Example 4, and the results are shown in FIG. 12 (marker is a polygonal line).

（実施例１０：官能試験）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きした。５００ｇの粗挽きした納豆と、３ｇの有胞子乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｓｐｏｒｏｇｅｎａｓ（三菱化学フーズ製）とを混合し、１０ｇのビタミンＣ（中国江蘇江山製薬製）、５０ｇの炭酸カルシウム（関東化学株式会社製）、及び蒸留水３００ｍＬを加え、４８時間３５℃で好気発酵した。発酵物を凍結乾燥し、粉末を得た。粉砕粉末のｐＨは、６．０５であり、納豆菌数は、１１．３×１０^９個／ｇであり、乳酸菌数は２．３×１０^９個／ｇであり、プロテアーゼ力価は乾燥粉末あたり１１，４００ｕ／ｇであった。２８歳〜７２歳の男女６名により、粉末の味、香り、及び色について官能試験を行い、結果を表１に示した。
味、及び香りについては、下記の評価基準により評価した。評価基準の点数が高いほど、好ましい味、及び香りであることを示す。 (Example 10: Sensory test)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a meat grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm. 500 g of coarsely ground natto and 3 g of Lactobacillus sporogenes (manufactured by Mitsubishi Kagaku Foods), a spore lactic acid bacterium, are mixed, 10 g of vitamin C (manufactured by Jiangsu Jiangshan Pharmaceutical Co., Ltd.), 50 g of calcium carbonate (manufactured by Kanto Kagaku Co., Ltd.), And 300 mL of distilled water were added, and aerobically fermented at 35° C. for 48 hours. The fermented product was freeze-dried to obtain a powder. The crushed powder had a pH of 6.05, a Bacillus natto count of 11.3×10 ⁹ cells/g, a lactic acid bacterium count of 2.3×10 ⁹ cells/g, and a protease titer of dry powder. It was 11,400 u/g. A sensory test was conducted on the taste, aroma, and color of the powder by 6 men and women aged 28 to 72, and the results are shown in Table 1.
The taste and aroma were evaluated according to the following evaluation criteria. The higher the evaluation standard score, the more preferable the taste and the scent.

−−評価基準−−
味
１：苦味
２：納豆の味
３：酸味、マイルドな味 ---Evaluation criteria---
Taste 1: Bitterness 2: Natto taste 3: Acidity, mild taste

−−評価基準−−
香り
１：納豆臭
２：納豆臭が少ない、発酵臭
３：納豆臭がしない ---Evaluation criteria---
Aroma 1: Natto odor 2: Little natto odor, fermented odor 3: No natto odor

（比較例３：官能試験）
納豆（株式会社豆紀製）を挽肉機（ＨＥＬＴＨＹＭＩＮＣＥＲ貝印株式会社製）、及び径４ｍｍのディスクを用いて粗挽きし、凍結乾燥した。粉砕粉末のｐＨは、６．５７であり、納豆菌数は、８．３×１０^９個／ｇであり、プロテアーゼ力価は１３，０００ｕ／ｇであった。２８歳〜７２歳の男女６名により、粉末の味、香り、及び色について官能試験を行い、結果を表１に示した。
味、及び香りについては、実施例１０と同様の評価基準により評価した。 (Comparative Example 3: Sensory test)
Natto (manufactured by Mame Co., Ltd.) was roughly ground using a grinder (HELTHY MINCER Kaijin Co., Ltd.) and a disc having a diameter of 4 mm, and freeze-dried. The ground powder had a pH of 6.57, a Bacillus natto count of 8.3×10 ⁹ cells/g, and a protease titer of 13,000 u/g. A sensory test was conducted on the taste, aroma, and color of the powder by 6 men and women aged 28 to 72, and the results are shown in Table 1.
The taste and aroma were evaluated according to the same evaluation criteria as in Example 10.

図１〜図３の結果から、納豆と乳酸菌とを混合して発酵することにより、大豆と乳酸菌とを混合して発酵した場合と比較して高濃度の乳酸菌の生菌を含有する食品が製造できることが分かった。更に、炭酸カルシウム及びビタミンＣを加えて発酵することにより、乳酸菌の生菌数が高くなることが分かった。
図４〜図５の結果から、納豆と酪酸菌とを混合して発酵することにより、納豆の生菌、及び高濃度の酪酸菌の生菌を含有する食品が製造できることが分かった。
図６〜７の結果から、納豆と麹菌とを混合して発酵することにより、納豆の生菌、及び高濃度の麹菌の生菌を含有する食品が製造できることが分かった。
図８〜図１０の結果から、納豆と、少なくとも２種類の乳酸菌とを混合して発酵することにより、納豆の生菌、及び高濃度の乳酸菌の生菌を含有する食品が製造できることが分かった。また、ビタミンＣを加えて発酵することにより、乳酸菌の比率が増加することが分かった。
図１１〜図１２の結果から、納豆と、乳酸菌とを混合して嫌気発酵することにより、高いプロテアーゼ活性を有し、納豆の生菌、及び高濃度の乳酸菌の生菌を含有する食品が製造できることが分かった。また、炭酸カルシウムを加えることで、乳酸菌の増殖が長期に維持されることが分かった。
表１の結果から、本発明の食品の製造方法により製造される食品は、好ましい味、及び香りを有することが分かった。
したがって、本発明の、食品の製造方法は、格別顕著な効果を奏するものである。

From the results of FIGS. 1 to 3, by mixing and fermenting natto and lactic acid bacteria, a food containing a high concentration of viable lactic acid bacteria is produced as compared with the case of mixing and fermenting soybeans and lactic acid bacteria. I knew I could do it. Further, it was found that the number of viable lactic acid bacteria was increased by adding calcium carbonate and vitamin C and performing fermentation.
From the results of FIGS. 4 to 5, it was found that by mixing and fermenting natto and butyric acid bacteria and fermenting, a food containing natto viable bacteria and a high concentration of butyric acid viable bacteria can be produced.
From the results of FIGS. 6 to 7, it was found that by mixing and fermenting natto and Aspergillus oryzae, a food containing natto viable bacteria and a high concentration of aspergillus viable bacteria can be produced.
From the results of FIGS. 8 to 10, it was found that by mixing natto and at least two kinds of lactic acid bacteria and fermenting them, a food containing natto viable bacteria and a high concentration of lactic acid bacterium viable bacteria can be produced. .. Further, it was found that the ratio of lactic acid bacteria was increased by adding vitamin C and fermenting.
From the results of FIGS. 11 to 12, by mixing natto and lactic acid bacteria and performing anaerobic fermentation, a food product having high protease activity and containing live natto bacteria and high-concentration lactic acid bacterium live bacteria is produced. I knew I could do it. It was also found that the addition of calcium carbonate maintained the growth of lactic acid bacteria for a long period of time.
From the results in Table 1, it was found that the food produced by the method for producing food of the present invention has a preferable taste and aroma.
Therefore, the method for producing food according to the present invention has a particularly remarkable effect.

Claims

Bacillus natto-containing material, lactic acid bacterium , and at least one of butyric acid bacterium are mixed, and the fermentation step of fermenting the natto bacterium-containing material with at least one of the lactic acid bacterium and butyric acid bacterium ,
A method for producing a food, wherein calcium carbonate and vitamin C are added in the fermentation step.

The production method according to claim 1, wherein the lactic acid bacterium contains at least two types of lactic acid bacteria.

The method according to any one of claims 1 to 2, wherein the concentrations of the Bacillus natto, lactic acid bacteria , and butyric acid bacteria in the food satisfy at least one of the following:
The concentration of the Bacillus natto is 1×10 ⁸ viable bacteria/g or more,
The concentration of the lactic acid bacteria, in live bacteria, is 1 × 10 ⁸ cells / g or more, or the concentration of the butyric acid bacteria, in live bacteria, Ru der 1 × 10 ⁸ cells / g or more.

The method according to any one of claims 1 to 3, wherein the protease titer in the food is 1,000 u/g or more.