JP6792781B2 - Fermented seasoning with high antihypertensive peptide content - Google Patents
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Description
本発明は、降圧ペプチドであるGly−Tyr及びSer−Tyrを高含有する発酵調味料及びその製造方法に関する。 The present invention relates to a fermented seasoning containing a high amount of the antihypertensive peptides Gly-Tyr and Ser-Tyr, and a method for producing the same.
近年、ペプチドが有する様々な生理機能が明らかとなってきている。例えば、Gly−Tyr及びSer−Tyrは、アンジオテンシン変換酵素阻害作用及び血圧降下作用を有することが知られている(非特許文献1)。よって、Gly−TyrやSer−Tyr(以下、降圧ペプチドGly−Tyr及び降圧ペプチドSer−Tyrと記載することがある)の効率的な摂取は、高血圧症や動脈硬化症などの改善に有効であるといえる。 In recent years, various physiological functions of peptides have been clarified. For example, Gly-Tyr and Ser-Tyr are known to have angiotensin converting enzyme inhibitory action and blood pressure lowering action (Non-Patent Document 1). Therefore, efficient ingestion of Gly-Tyr and Ser-Tyr (hereinafter, may be referred to as antihypertensive peptide Gly-Tyr and antihypertensive peptide Ser-Tyr) is effective in improving hypertension and arteriosclerosis. It can be said that.
一方、醤油は古くから使用されてきた調味料であり、大豆と小麦を麹菌の産生する酵素が分解することにより、豊かな旨味が醸し出される。この旨味は、原料のタンパク質にプロテアーゼが作用してペプチドとして可溶化され、さらにこれらのペプチドにペプチダーゼが作用してグルタミン酸をはじめとしたアミノ酸に分解することにより生じる。これまで降圧ペプチドGly−Tyr及び降圧ペプチドSer−Tyrを、醤油に含有させる試みとしては、諸味液汁中のロイシンアミノペプチダーゼ−I及びロイシンアミノペプチダーゼ−IIの活性を制御することによって、諸味液汁中に可溶化したペプチドの分解を抑制する方法(特許文献1)、低プロテアーゼ醤油麹の利用と該醤油麹の加温消化を組み合わせる方法(特許文献2)が報告されている。しかしながら、より簡単な方法にて、醤油諸味中に生成蓄積される降圧ペプチドGly−Tyr及び降圧ペプチドSer−Tyrを増加させた血圧降下能の高い発酵調味料の開発が望まれるところである。 On the other hand, soy sauce is a seasoning that has been used for a long time, and the enzyme produced by Jiuqu decomposes soybeans and wheat to produce a rich flavor. This umami is produced by the action of protease on the raw material protein to solubilize it as a peptide, and the action of peptidase on these peptides to decompose them into amino acids such as glutamic acid. So far, as an attempt to contain the antihypertensive peptide Gly-Tyr and the antihypertensive peptide Ser-Tyr in soy sauce, the activities of leucine aminopeptidase-I and leucine aminopeptidase-II in the soy sauce juice have been controlled. A method of suppressing the decomposition of solubilized peptides (Patent Document 1) and a method of combining the use of low protease soy sauce koji with warm digestion of the soy sauce koji have been reported (Patent Document 2). However, it is desired to develop a fermented seasoning having a high blood pressure lowering ability by increasing the antihypertensive peptide Gly-Tyr and the antihypertensive peptide Ser-Tyr produced and accumulated in soy sauce moromi by a simpler method.
本発明の目的は、降圧ペプチドであるGly−Tyr及びSer−Tyrを従来品よりも多く含有し、血圧降下効果のより高い発酵調味料を提供することにある。 An object of the present invention is to provide a fermented seasoning containing more antihypertensive peptides Gly-Tyr and Ser-Tyr than conventional products and having a higher blood pressure lowering effect.
本発明者らは、上記の課題を解決するため鋭意研究を重ねた結果、原料大豆中のβ−コングリシニンとグリシニンの合計を100重量部とした時にβ−コングリシニンが50重量部以上となる大豆を醤油麹に混合して食塩水に仕込んで諸味を調製し、該諸味を25℃〜60℃で発酵させることにより得られる発酵調味料が、驚くべきことに通常の大豆を原料に用いた場合よりも降圧ペプチドであるGly−Tyr及びSer−Tyrの含量が顕著に増加することを見出し、本発明に至った。 As a result of diligent research to solve the above problems, the present inventors have obtained soybeans having β-conglycinin of 50 parts by weight or more when the total of β-conglycinin and glycinin in the raw material soybean is 100 parts by weight. The fermented seasoning obtained by mixing with soy sauce koji and adding it to salt solution to prepare moromi and fermenting the moromi at 25 ° C to 60 ° C is surprisingly better than when ordinary soybeans are used as raw materials. Also found that the contents of the antihypertensive peptides Gly-Tyr and Ser-Tyr were significantly increased, leading to the present invention.
即ち、本発明は以下の発明を包含する。
(1)原料大豆中のβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの含有率が50w/w%以上であり、かつ、Gly−Tyr及びSer−Tyrを含有することを特徴とする、発酵調味料。
(2)前記原料大豆が、掛け原料である、(1)に記載の発酵調味料。
(3)前記原料大豆が、11Sグロブリン欠失又は低下大豆を含む、(1)又は(2)に記載の発酵調味料。
(4)前記原料大豆が、単一の又は複数の品種の大豆を含む、(1)〜(3)のいずれかに記載の発酵調味料。
(5)総窒素濃度1w/w%あたり、Gly−Tyrを70μg/ml以上、Ser−Tyrを100μg/ml以上含有する、(1)〜(4)のいずれかに記載の発酵調味料。
(6)血圧降下用発酵調味料である、(1)〜(5)のいずれかに記載の発酵調味料。
(7)原料大豆と麹菌培養物と食塩水を混和して諸味を調製する工程と、該諸味を25〜60℃で発酵させる工程を含む、Gly−Tyr及びSer−Tyrを含有する発酵調味料の製造方法であって、該原料大豆中のβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの含有率が50w/w%以上である、発酵調味料の製造方法。
That is, the present invention includes the following inventions.
(1) Fermentation characterized in that the content of β-conglycinin with respect to the total amount of β-conglycinin and glycinein in the raw material soybean is 50 w / w% or more, and Gly-Tyr and Ser-Tyr are contained. seasoning.
(2) The fermented seasoning according to (1), wherein the raw material soybean is a hanging material.
(3) The fermented seasoning according to (1) or (2), wherein the raw material soybean contains 11S globulin-deficient or reduced soybean.
(4) The fermented seasoning according to any one of (1) to (3), wherein the raw material soybean contains a single or a plurality of varieties of soybean.
(5) The fermented seasoning according to any one of (1) to (4), which contains 70 μg / ml or more of Gly-Tyr and 100 μg / ml or more of Ser-Tyr per 1 w / w% of total nitrogen concentration.
(6) The fermented seasoning according to any one of (1) to (5), which is a fermented seasoning for lowering blood pressure.
(7) Fermented seasoning containing Gly-Tyr and Ser-Tyr, which comprises a step of mixing raw soybeans, a culture of aspergillus and salt solution to prepare various flavors, and a step of fermenting the various flavors at 25 to 60 ° C. The method for producing a fermented seasoning, wherein the content of β-conglycinin with respect to the total amount of β-conglycinin and glycinin in the raw material soybean is 50 w / w% or more.
本発明によれば、降圧ペプチドGly−Tyr及び降圧ペプチドSer−Tyrを従来品よりも顕著に増加した発酵調味料が提供される。 According to the present invention, there is provided a fermented seasoning in which the antihypertensive peptide Gly-Tyr and the antihypertensive peptide Ser-Tyr are significantly increased as compared with the conventional product.
以下、本発明を詳細に説明する。
本発明の発酵調味料は、原料大豆中のβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの含有率が50w/w%以上であり、かつ、Gly−Tyr及びSer−Tyrを含有することを特徴とする。
Hereinafter, the present invention will be described in detail.
The fermented seasoning of the present invention has a β-conglycinin content of 50 w / w% or more with respect to the total amount of β-conglycinin and glycine in the raw soybean, and contains Gly-Tyr and Ser-Tyr. It is a feature.
(原料大豆)
大豆の貯蔵タンパク質は、沈降係数(S)により2S、7S、11S及び15Sグロブリンに分類され、また、免疫学的命名法により、グリシニン、α−、β−、γ−コングリシニンに分類される。これらのうち、7Sグロブリン(β−コングリシニン)と、11Sグロブリン(グリシニン)が大豆の貯蔵タンパク質の大部分を占め、7Sグロブリンは、α、α’、及びβサブユニットから構成されており、一方、11Sグロブリンは酸性ポリペプチド(A)と塩基性ポリペプチド(B)がジスルフィド結合(S−S結合)で重合し、A1aB2、A1bB1b、A2B1a、A5A4B3及びA3B4サブユニットから構成されている。本発明の発酵調味料に用いる原料大豆は、β−コングリシニン含有率がグリシニンの含有率よりも高い大豆をいい、具体的には、原料大豆中のβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの含有率が50w/w%以上であればよいが、70w/w%以上が好ましく、80w/w%以上がより好ましく、90w/w%以上がさらに好ましく、95w/w%以上が最も好ましい。本発明の原料大豆として用いるβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの量が50w/w%以上となる大豆(以下、「β−コングリシニン高含有大豆」という)としては、11Sグロブリン欠失又は低下大豆、例えば、「ななほまれ」、「関東133号」等のグリシニン(11Sグロブリン)の一部または全てを欠失させてβ−コングリシニン(7Sグロブリン)含量を増加させた大豆、施肥等の栽培条件でβ−コングリシニン(7Sグロブリン)含量を相対的に増加させた大豆等があげられるが、これらに限定はされない。
(Ingredient soybean)
Soybean storage proteins are classified into 2S, 7S, 11S and 15S globulin by sedimentation coefficient (S) and glycinin, α-, β- and γ-conglycinin by immunological nomenclature. Of these, 7S globulin (β-conglycinin) and 11S globulin (glycinin) make up the majority of soybean storage proteins, and 7S globulin is composed of α, α', and β subunits, while 11S globulin is composed of A1aB2, A1bB1b, A2B1a, A5A4B3 and A3B4 subunits obtained by polymerizing an acidic polypeptide (A) and a basic polypeptide (B) with a disulfide bond (SS bond). The raw material soybean used in the fermented seasoning of the present invention refers to soybean having a β-conglycinin content higher than that of glycinin, and specifically, β-conglycinin with respect to the total amount of β-conglycinin and glycinin in the raw material soybean. The content of soybean may be 50 w / w% or more, but 70 w / w% or more is preferable, 80 w / w% or more is more preferable, 90 w / w% or more is further preferable, and 95 w / w% or more is most preferable. 11S globulin-deficient soybean having an amount of β-conglycinin of 50 w / w% or more based on the total amount of β-conglycinin and glycinin used as the raw material soybean of the present invention (hereinafter referred to as “β-conglycinin high content soybean”) Or reduced soybeans, for example, soybeans in which β-conglycinin (7S globulin) content is increased by deleting part or all of glycinin (11S globulin) such as "Nanahomare" and "Kanto 133", fertilization, etc. Examples thereof include soybeans having a relatively increased β-conglycinin (7S globulin) content under the cultivation conditions of, but are not limited thereto.
上記の大豆品種は、単一の品種を用いてよく、複数の品種をブレンドして用いてもよい。また、原料大豆におけるβ−コングリシニンの含有率が上記の範囲であれば、β−コングリシニン高含有大豆に一般の大豆が混合されていてもよい。さらに、分離精製したβ−コングリシニン(例えば、不二製油社製「リポフ」)を原料大豆と混合することにより、原料大豆中のβ−コングリシニンとグリシニンの合計量に対するβ−コングリシニンの含有率を増加させてもよい。 As the above soybean varieties, a single variety may be used, or a plurality of varieties may be blended and used. Further, as long as the content of β-conglycinin in the raw material soybean is within the above range, general soybean may be mixed with the soybean containing high β-conglycinin. Furthermore, by mixing separated and purified β-conglycinin (for example, “Lipov” manufactured by Fuji Oil Co., Ltd.) with the raw material soybean, the content of β-conglycinin in the raw material soybean with respect to the total amount of β-conglycinin and glycinin is increased. You may let me.
原料大豆は、そのままあるいは割砕、粉砕してから、適宜タンパク質変性処理を施し、用いることができる。タンパク質変性処理としては、工業的に広く行われている加圧蒸煮や膨化処理が好ましい。 The raw material soybean can be used as it is, or after being crushed and crushed, and then appropriately subjected to protein denaturation treatment. As the protein denaturation treatment, pressure steaming and swelling treatment, which are widely performed industrially, are preferable.
本発明の発酵調味料の製造において、上記の原料大豆は、諸味調製時に麹に混合する掛け原料として用いることが好ましいが、製麹原料として用いることもできる。 In the production of the fermented seasoning of the present invention, the above-mentioned raw material soybean is preferably used as a hanging material to be mixed with the jiuqu at the time of preparing various flavors, but it can also be used as a raw material for making koji.
(麹菌培養物)
麹菌培養物としては、醤油醸造で広く一般的に用いられている「醤油麹」を使用することができる。具体的には、大豆、脱脂加工大豆、小麦グルテンなどを加熱変性して得られた蛋白質原料と、麦類(小麦、大麦、裸麦、はと麦)、米類などを加熱変性して得られた澱粉質原料を含む固体培地に種麹菌を接種し、25〜45℃で、12〜240時間培養(製麹)して得ることができる。麦類の加熱変性は炒熬割砕によって行うことが好ましく、米類の加熱変性は蒸煮又は炊飯によって行うことが好ましい。ここで、蛋白質原料として大豆を用いる場合、普通の大豆を用いてもよく、また、上記のβ−コングリシニン高含有大豆を用いてよい。また、麹菌培養物としては、発酵調味料の製造に使用されている「液体培養麹」を使用することもできる。具体的には、上記の蛋白質原料と澱粉質原料を1〜5w/v%含む液体培地に種麹菌を接種し、25〜45℃で、24〜120時間培養(製麹)して得ることができる。ここで、種麹菌には、アスペルギルス・ソーヤ(Aspergillus sojae)、アスペルギルス・オリーゼ(Aspergillus oryzae)等が用いられる。
(Jiuqu culture)
As the aspergillus culture, "soy sauce koji" widely and generally used in soy sauce brewing can be used. Specifically, it is obtained by heat-modifying soybeans, defatted soybeans, wheat gluten, etc., and heat-modifying wheat (wheat, barley, bare wheat, hato wheat), rice, etc. It can be obtained by inoculating a solid medium containing a starchy raw material with seed aspergillus and culturing (jiuqu) at 25 to 45 ° C. for 12 to 240 hours. The heat denaturation of wheat is preferably performed by stir-fry crushing, and the heat denaturation of rice is preferably performed by steaming or cooking rice. Here, when soybean is used as a protein raw material, ordinary soybean may be used, or the above-mentioned β-conglycinin-rich soybean may be used. Further, as the aspergillus culture, "liquid cultured aspergillus" used for producing fermented seasonings can also be used. Specifically, it can be obtained by inoculating the seed aspergillus in a liquid medium containing 1 to 5 w / v% of the above protein raw material and starchy raw material and culturing (jiuqu) at 25 to 45 ° C. for 24 to 120 hours. it can. Here, Aspergillus sojae, Aspergillus oryzae, and the like are used as the seed aspergillus.
上記蛋白質原料と澱粉質原料との配合比(蛋白質原料:澱粉質原料)は、特に限定はされないが、例えば、濃口醤油、淡口醤油で用いられている配合比である30:70〜70:30が好ましく、40:60〜60:40がより好ましい。また、蛋白質原料と澱粉質原料の混和物の水分は、35〜50w/w%が好ましく、40〜45w/w%がより好ましい。 The blending ratio of the protein raw material and the starch raw material (protein raw material: starch raw material) is not particularly limited, but is, for example, the blending ratio used in dark soy sauce and light soy sauce from 30:70 to 70:30. Is preferable, and 40:60 to 60:40 is more preferable. The water content of the mixture of the protein raw material and the starchy raw material is preferably 35 to 50 w / w%, more preferably 40 to 45 w / w%.
(諸味の調製)
次に、麹菌培養物、掛け原料としてβ−コングリシニン高含有大豆、及び食塩水を混和して(仕込み)、諸味を調製する。仕込みの条件としては、β−コングリシニン高含有大豆と麹菌培養物を5:95〜95:5、好ましくは10:90〜80:20の比で混合したものに、重量比で30〜500%、好ましくは100〜300%の食塩水を加える。食塩水の濃度は、諸味液汁の食塩濃度として0.1〜20w/v%が好ましく、6〜18w/v%がより好ましい。
(Preparation of moromi)
Next, the aspergillus culture, soybeans high in β-conglycinin as a hanging material, and saline solution are mixed (prepared) to prepare moromi mash. The conditions for the preparation were a mixture of soybeans high in β-conglycinin and a culture of Jiuqu at a ratio of 5:95 to 95: 5, preferably 10:90 to 80:20, and a weight ratio of 30 to 500%. Preferably 100-300% saline is added. The concentration of the salt solution is preferably 0.1 to 20 w / v%, more preferably 6 to 18 w / v%, as the salt concentration of the moromi juice.
(諸味の発酵・熟成)
次に、調製した諸味を発酵・熟成させる。諸味の発酵・熟成は、諸味を温度制御が可能な容器に入れ、25〜60℃、好ましくは30〜55℃で、6〜720時間、好ましくは12〜240時間保持することより行う。この間、攪拌翼や圧縮空気等によって適宜諸味を攪拌してもよい。
(Fermentation and aging of various flavors)
Next, the prepared mash is fermented and aged. Fermentation and aging of moromi are carried out by placing the moromi in a container capable of controlling the temperature and holding the moromi at 25 to 60 ° C., preferably 30 to 55 ° C. for 6 to 720 hours, preferably 12 to 240 hours. During this time, various tastes may be appropriately stirred with a stirring blade, compressed air, or the like.
上記の発酵・熟成後の諸味は、醤油諸味様の形態であり、そのままで本発明の発酵調味料として用いることできるが、必要に応じ固形分を除去することにより、降圧ペプチドを含む液体状の発酵調味料を得ることもできる。 The above-mentioned fermented and aged moromi mash is in the form of soy sauce mash, and can be used as it is as the fermented seasoning of the present invention. You can also get fermented seasonings.
また、上記発酵・熟成後の諸味に醤油乳酸菌を接種して十分に乳酸菌を繁殖させ、乳酸発酵を行うか、上記諸味に醤油酵母を接種して十分に酵母を繁殖させ、アルコール発酵を行うか、又は、該乳酸発酵及び該アルコール発酵をこの順で行い、香味をさらに向上させたものを、本発明の発酵調味料として用いてもよい。 In addition, whether to inoculate the above-mentioned various flavors after fermentation and aging with soy sauce lactic acid bacteria to sufficiently propagate the lactic acid bacteria and perform lactic acid fermentation, or to inoculate the above-mentioned various flavors with soy sauce yeast to sufficiently propagate the yeast and perform alcoholic fermentation. Alternatively, the lactic acid fermentation and the alcoholic fermentation are carried out in this order to further improve the flavor, which may be used as the fermentation seasoning of the present invention.
乳酸発酵は、諸味中に0.5w/v%以上の乳酸を生成させるまで行うことが好ましい。乳酸発酵に使用できる乳酸菌としては、テトラジェノコッカス・ハロフィルス(Tetragenococcus halophilus)等の耐塩性醤油乳酸菌が挙げられる。乳酸発酵は、諸味に対し、1×103〜1×107個/g諸味となるように乳酸菌を添加し、次いで諸味品温を20〜35℃、好ましくは25〜30℃で、3日以上、好ましくは5〜10日発酵させる。発酵を旺盛にするために、適宜攪拌や通気を行ってもよい。 Lactic acid fermentation is preferably carried out until 0.5 w / v% or more of lactic acid is produced in the mash. Examples of lactic acid bacteria that can be used for lactic acid fermentation include salt-tolerant soy sauce lactic acid bacteria such as Tetragenococcus halophilus. In the lactic acid fermentation, lactic acid bacteria are added so that the moromi is 1 × 10 3 to 1 × 10 7 pieces / g moromi, and then the moromi temperature is 20 to 35 ° C, preferably 25 to 30 ° C for 3 days. As mentioned above, it is preferably fermented for 5 to 10 days. In order to make the fermentation vigorous, stirring and aeration may be performed as appropriate.
乳酸発酵開始時の諸味は、通常pH5.8〜6.2であるが、次第に溶解(加水分解)して、原料溶出及び乳酸発酵が開始され、pHが4.6〜5.3に低下し、酵母の増殖に好適な諸味となる。 The various tastes at the start of lactic acid fermentation are usually pH 5.8 to 6.2, but gradually dissolve (hydrolyze), elution of raw materials and lactic acid fermentation are started, and the pH drops to 4.6 to 5.3. , Suitable for yeast growth.
本発明においてアルコール発酵に用いられる酵母としては、10〜20w/v%の食塩高濃度の諸味中でも旺盛に増殖できる耐塩性醤油酵母が好ましく、例えば、チゴサッカロマイセス・ルーキシー(Zygosaccharomyces rouxii)、チゴサッカロマイセス・バイリー(Zygosaccharomyces bailli)などのチゴサッカロマイセス(Zygosaccharomyces)属の酵母、キャンディダ・バーサチルス(Candida versatilus)、キャンディダ・エチェリシ(Candida etchellsii)、キャンディダ・トロピカリス(Candida tropicalis)等のキャンディダ(Candida)属の酵母が挙げられる。 As the yeast used for alcohol fermentation in the present invention, salt-tolerant soybean yeast that can grow vigorously even in various tastes having a high salt concentration of 10 to 20 w / v% is preferable, and for example, Zygosaccharomyces rouxii, Zygosaccharomyces rouxii, Zygosaccharomyces rouxii, etc. Yeasts of the genus Zygosaccharomyces such as Bailli (Zygosaccharomyces bailli), Candida such as Candida versatilus, Candida etchellsii, Candida tropicalis Examples include yeast of the genus.
酵母は、諸味1g当たり1×103〜1×107個となるように、諸味に予め培養しておいた上記酵母を添加することが好ましい。また、予め培養しておいた酵母を人為的に添加することなく、諸味に空中落下又は醤油麹から自然混入した酵母、あるいは醤油諸味が仕込まれた発酵槽等に棲息する耐塩性醤油酵母(野生型酵母)を利用してもよい。 It is preferable to add the yeast previously cultivated to the moromi so that the number of yeasts is 1 × 10 3 to 1 × 10 7 per 1 g of the moromi. In addition, yeast that has been dropped into the air or naturally mixed from soy sauce moromi without artificially adding yeast that has been cultivated in advance, or salt-resistant soy sauce yeast that lives in fermenters or the like in which soy sauce moromi is charged (wild). Type yeast) may be used.
アルコール発酵は、酵母菌を添加後、諸味品温を20〜35℃にて、アルコールが0.5〜4w/v%、好ましくは3〜4w/v%程度となるのに十分な時間、例えば30〜90日間程度保持することにより行う。諸味中のエチルアルコール濃度は、培養時間によって調整することができる。また、酵母の発酵・熟成は、間歇的又は連続的に通気培養、攪拌培養、通気攪拌培養を行うなど、好気的条件下で実施することが好ましい。 In the alcoholic fermentation, after adding yeast, the temperature of the mash is 20 to 35 ° C., and the alcohol content is 0.5 to 4 w / v%, preferably about 3 to 4 w / v%, for a sufficient time, for example. This is done by holding for about 30 to 90 days. The ethyl alcohol concentration in the moromi can be adjusted by the culture time. Further, fermentation and aging of yeast are preferably carried out under aerobic conditions such as intermittent or continuous aeration culture, stirring culture, and aeration stirring culture.
アルコール発酵を終えた諸味はさらに静置し、酵母・乳酸菌による発酵を継続し、その後、熟成の程度は問わないが通常数週間から数ヵ月程度熟成させる。 After alcoholic fermentation, the moromi mash is allowed to stand still, and fermentation with yeast and lactic acid bacteria is continued. Then, the moromi is usually aged for several weeks to several months regardless of the degree of aging.
(圧搾・火入れ)
上記の発酵・熟成後の諸味は、常法により圧搾濾過した後、「火入れ」と呼ばれる加熱処理を行ない、オリ引き(清澄処理)し、容器に充填して製品化する。圧搾濾過は、ナイロンなどの合成繊維でできた濾布で諸味を包み、加圧する厚揚げ方式や、濾過板と圧搾板に張った濾布の中に諸味を入れ、圧縮空気等で加圧するフィルタープレス方式等の従来公知の方法を用いることができる。火入れは公知の醤油醸造で用いられている加熱条件で行えばよく、例えば、80〜85℃で20〜60分間、又は110〜120℃で5〜20秒間行う。オリ引き(清澄処理)の方法は、膜処理、珪藻土ろ過、遠心分離、凝集法、沈降法など従来公知の方法を用いることができる。
(Squeezing / burning)
The above-mentioned moromi mash after fermentation and aging is squeezed and filtered by a conventional method, then subjected to a heat treatment called "burning", mashed (clarified), filled in a container, and commercialized. Press filtration is a thick-fried method that wraps various flavors in a filter cloth made of synthetic fibers such as nylon and pressurizes them, or a filter that puts various flavors in a filter plate and a filter cloth stretched on a press plate and pressurizes with compressed air or the like. Conventionally known methods such as a press method can be used. The burning may be carried out under the heating conditions used in known soy sauce brewing, for example, at 80 to 85 ° C. for 20 to 60 minutes, or at 110 to 120 ° C. for 5 to 20 seconds. As the method of sedimentation (clarification treatment), conventionally known methods such as membrane treatment, diatomaceous earth filtration, centrifugation, agglutination method, and sedimentation method can be used.
本発明の発酵調味料は、降圧ペプチドであるGly−Tyr及びSer−Tyrを従来品よりも1.2倍以上含有する。具体的には、本発明の発酵調味料は、総窒素濃度1w/w%あたり、Gly−Tyrを70μg/ml以上、好ましくは80μg/ml以上、及びSer−Tyrを100μg/ml以上、好ましくは110μg/ml以上含有し、より優れた血圧降下作用が期待できる。 The fermented seasoning of the present invention contains 1.2 times or more of the antihypertensive peptides Gly-Tyr and Ser-Tyr as compared with the conventional product. Specifically, the fermented seasoning of the present invention has Gly-Tyr of 70 μg / ml or more, preferably 80 μg / ml or more, and Ser-Tyr of 100 μg / ml or more, preferably 100 μg / ml or more, per 1 w / w% of total nitrogen concentration. It contains 110 μg / ml or more and can be expected to have a more excellent blood pressure lowering effect.
また、本発明の発酵調味料は、上記降圧ペプチドを著量含有する一方、苦味がほとんど感じられず、まろやかで豊富な旨味・コクと、穏やかで優れた芳香を有し、調味料としておいしく摂取できるものである。 In addition, the fermented seasoning of the present invention contains a significant amount of the above-mentioned antihypertensive peptide, but has almost no bitterness, has a mellow and abundant umami and richness, and has a mild and excellent aroma, and is deliciously ingested as a seasoning. It can be done.
降圧ペプチドは、前記発酵調味料から濃縮あるいは分離精製することもできる。その方法としては、減圧濃縮、限外濾過、透析、各種クロマトグラフィー等が挙げられる。これらは、一般に広く用いられている方法である。 The antihypertensive peptide can also be concentrated or separated and purified from the fermented seasoning. Examples of the method include vacuum concentration, ultrafiltration, dialysis, and various types of chromatography. These are commonly used methods.
本発明の発酵調味料は、そのまま用いてもよいが、液状のまま、あるいは常法により凍結乾燥、噴霧乾燥、ドラムドライ乾燥などして、ペースト状、固形状、粉末状とした後、各種つゆ(麺つゆなど)、たれ(焼肉用たれなど)、ぽん酢、ソース、ドレッシング、スープなどの液体調味料に添加することで醤油含有調味料としても用いることができる。また、水産・畜肉製品(蒲鉾、ハム等)の食品に添加することで、血圧降下用飲食品として提供できる。 The fermented seasoning of the present invention may be used as it is, but it may be used as it is, or it may be made into a paste, solid or powder by freeze-drying, spray-drying, drum-drying or the like by a conventional method, and then various soy sauces. It can also be used as a soy sauce-containing seasoning by adding it to liquid seasonings such as (noodle soup, etc.), sauce (sauce for roasted meat, etc.), ponzu, sauce, dressing, soup, etc. In addition, it can be provided as a food or drink for lowering blood pressure by adding it to foods of marine products and livestock meat products (kamaboko, ham, etc.).
また、本発明の発酵調味料は、降圧ペプチドを著量含有するので、血圧降下によってよって病態が改善される疾患の予防及び/又は改善を目的とした血圧降下用発酵調味料として利用できる。上記疾患としては、例えば、高血圧症、血栓症、動脈硬化症、狭心症、心筋梗塞、脳梗塞などが挙げられるが、これらに限定はされない。本発明の血圧降下用発酵調味料は、医薬品以外で健康の維持や増進を目的として摂取できる食品、例えば、健康食品、機能性食品、保健機能食品、又は特別用途食品を含む意味で用いられる。健康食品には、栄養補助食品、健康補助食品、サプリメント等の名称で提供される食品を含む。保健機能食品は食品衛生法又は食品増進法により定義され、特定の保健の効果や栄養成分の機能、疾病リスクの低減などを表示できる、特定保健用食品及び栄養機能食品、ならびに科学的根拠に基づいた機能性について消費者庁長官に届け出た内容を表示できる機能性表示食品が含まれる。また特別用途食品には、特定の対象者や特定の疾患を有する患者に適する旨を表示する病者用食品、高齢者用食品、妊産婦用食品等が含まれる。 In addition, since the fermented seasoning of the present invention contains a significant amount of antihypertensive peptide, it can be used as a fermented seasoning for lowering blood pressure for the purpose of preventing and / or ameliorating a disease in which the pathological condition is improved by lowering blood pressure. Examples of the above-mentioned diseases include, but are not limited to, hypertension, thrombosis, arteriosclerosis, angina, myocardial infarction, and cerebral infarction. The fermented seasoning for lowering blood pressure of the present invention is used to include foods other than pharmaceuticals that can be ingested for the purpose of maintaining or promoting health, such as health foods, functional foods, health functional foods, or special purpose foods. Health foods include foods provided under the names of dietary supplements, dietary supplements, supplements and the like. Health functional foods are defined by the Food Hygiene Law or the Food Promotion Law, and are based on specified health foods and nutritionally functional foods that can display specific health effects, functions of nutritional components, reduction of disease risk, etc., and scientific evidence. Includes foods with functional claims that can display the contents notified to the Commissioner of the Consumer Affairs Agency. In addition, special-purpose foods include foods for the sick, foods for the elderly, foods for pregnant women, etc. that indicate that they are suitable for a specific target person or a patient having a specific disease.
以下、実施例によって本発明を更に具体的に説明するが、これらの実施例は本発明を限定するものでない。 Hereinafter, the present invention will be described in more detail with reference to Examples, but these Examples do not limit the present invention.
(参考例)β−コングリシニン高含有大豆の育種及びβ−コングリシニン量の測定
(1) β−コングリシニン高含有大豆の育種
β-コングリシニン高含有大豆の育成は、文献(Yagasaki, K., N. Kaizuma and K. Kitamura(1996)Inheritance of glycinin subunits and characterization of glycinin molecules lacking the subunits in soybean (Glycine max (L.) Merr.). Breeding Science 46(1):11-15.)に記載の方法に従い、以下のとおり行った。11S欠失大豆「ななほまれ」を遺伝子供与親とし、普通大豆「サチユタカ」を戻し交雑親として戻し交雑を行った。得られたBC1F1種子は圃場で栽培し、BC1F2種子を収穫した。収穫したBC1F2種子からSDS-ポリアクリルアミドゲル電気泳動で11Sの3つのサブユニットを欠失した個体を選抜し、再度「サチユタカ」に戻し交雑を行った。この操作を2回繰り返し、得られたBC3F1種子を圃場で栽培してBC3F2種子を収穫した。このBC3F2種子からSDS-ポリアクリルアミドゲル電気泳動で11Sの3つのサブユニットを欠失した個体を選抜し、遺伝的な固定化を図るために、BC3F3まで世代を進めた。BC3F3世代以降は農業特性に優れた系統を選抜することで、11Sを欠失しβ-コングリシニンを高含有する大豆を作出した。
(Reference example) Breeding of soybeans high in β-conglycinin and measurement of β-conglycinin content
(1) Breeding of β-conglycinin-rich soybeans Breeding of β-conglycinin-rich soybeans is described in the literature (Yagasaki, K., N. Kaizuma and K. Kitamura (1996) Inheritance of glycinin subunits and characterization of glycinin molecules lacking the subunits. According to the method described in in soybean (Glycine max (L.) Merr.). Breeding Science 46 (1): 11-15.), The procedure was performed as follows. Backcrossing was performed with the 11S-deficient soybean "Nanahomare" as the gene donor and the normal soybean "Sachiyutaka" as the backcross parent. The obtained BC 1 F 1 seeds were cultivated in the field, and BC 1 F 2 seeds were harvested. Individuals lacking the three subunits of 11S were selected from the harvested BC 1 F 2 seeds by SDS-polyacrylamide gel electrophoresis, and then returned to "Sachiyutaka" for crossing. This operation was repeated twice, and the obtained BC 3 F 1 seeds were cultivated in the field to harvest BC 3 F 2 seeds. Individuals lacking the three subunits of 11S by SDS-polyacrylamide gel electrophoresis were selected from these BC 3 F 2 seeds, and the generation was advanced to BC 3 F 3 in order to achieve genetic immobilization. From the BC 3 F 3 generation onward, soybeans lacking 11S and having a high β-conglycinin content were produced by selecting strains with excellent agricultural characteristics.
(2) β−コングリシニン量の測定
β−コングリシニン量の測定は以下の通り行った。一般的な外国産大豆1種、一般的な国産大豆4種(A,B,C,D)及び(1)で得られたβ−コングリシニン高含有大豆をそれぞれミルサーIFM−620DG(岩谷産業社製)を用いて挽いた。挽いた大豆粉末に10倍量の純水を加え、ポリトロンホモジナイザーによる破砕を行った。4,000Gで15分間遠心し、上清の水溶性画分サンプルを得た。純水で100倍に希釈したサンプル10μLをSDS−PAGEに供した。SDS−PAGEは定法に従って行った。SDS−PAGE後のゲルはQuick−CBBキット(和光純薬工業社製)を用いて染色し、スキャナーを用いて画像化した。染色したゲルの画像を図1に示し、矢印で示したバンドは上からそれぞれβ−コングリシニンα及びα’サブユニット、β−コングリシニンβサブユニット、グリシニン酸性(acidic)IIbサブユニット、グリシニン酸性(acidic)I及びIIaサブユニット、グリシニン塩基性(basic)サブユニットを示す。各バンドの強度を、画像解析ソフトウェアImageJを用いて数値化し、β−コングリシニンのバンドの合計が、β−コングリシニン及びグリシニンのバンドの合計に占める割合をβ−コングリシニン率として算出した。結果を表1に示す。
(2) Measurement of β-conglycinin amount The β-conglycinin amount was measured as follows. Milcer IFM-620DG (manufactured by Iwatani Corp.) contains soybeans with high β-conglycinin content obtained from one general foreign soybean, four general domestic soybeans (A, B, C, D) and (1). ) Was used for grinding. Ten times the amount of pure water was added to the ground soybean powder, and the mixture was crushed with a polytron homogenizer. Centrifugation at 4,000 G for 15 minutes gave a water-soluble fraction sample of the supernatant. 10 μL of a sample diluted 100-fold with pure water was subjected to SDS-PAGE. SDS-PAGE was performed according to a conventional method. The gel after SDS-PAGE was stained with a Quick-CBB kit (manufactured by Wako Pure Chemical Industries, Ltd.) and imaged with a scanner. Images of the stained gel are shown in FIG. 1, and the bands indicated by arrows are β-conglycinin α and α'subunits, β-conglycinin β subunit, glycinin acidic IIb subunit, and glycinin acidic (acidic), respectively, from the top. ) I and IIa subunits, glycinin basic subunits. The intensity of each band was quantified using the image analysis software ImageJ, and the ratio of the total of the β-conglycinin bands to the total of the β-conglycinin and glycinin bands was calculated as the β-conglycinin ratio. The results are shown in Table 1.
(実施例1、比較例1)ペプチド含有発酵調味料の製造
加熱変性した大豆(一般的な大豆)と小麦を等量含む固体培地に、麹菌アスペルギルス・ソーヤの胞子を添加して、25〜40℃で72時間製麹することにより醤油麹を得た。次に、加熱変性した大豆(β−コングリシニン高含有大豆、国産大豆A)260g、醤油麹60g、食塩50g、水200mLを三角フラスコ中で混合し、50℃、140rpmで3日間振とう攪拌して諸味を得た。ここで醤油麹に混合する大豆として、β−コングリシニン高含有大豆を用いたものを実施例1、国産大豆Aを用いたものを比較例1とする。諸味調製後、濾紙による濾過で諸味から不溶性固形分を除去し、清澄液を得た。さらに、清澄液を85℃で30分間加熱し、酵素失活と殺菌を行った。10,000g、10分間遠心を行い、上清を採取して液体状のペプチド含有発酵調味料を得た。
(Example 1, Comparative Example 1) Production of fermented seasoning containing peptide 25-40 by adding spores of Aspergillus oryzae aspergillus oryzae to a solid medium containing equal amounts of heat-modified soybean (general soybean) and wheat. Soy sauce koji was obtained by making koji at ° C for 72 hours. Next, 260 g of heat-modified soybean (soybean containing high β-conglycinin, domestic soybean A), 60 g of soy sauce koji, 50 g of salt, and 200 mL of water were mixed in an Erlenmeyer flask, and stirred at 50 ° C. and 140 rpm for 3 days. I got various tastes. Here, as the soybeans to be mixed with the soy sauce koji, those using β-conglycinin-rich soybeans are referred to as Example 1, and those using domestic soybean A are referred to as Comparative Example 1. After preparing the mash, insoluble solids were removed from the mash by filtration through a filter paper to obtain a clarified solution. Further, the clarified solution was heated at 85 ° C. for 30 minutes to perform enzyme deactivation and sterilization. The mixture was centrifuged at 10,000 g for 10 minutes, and the supernatant was collected to obtain a liquid peptide-containing fermented seasoning.
得られたペプチド含有発酵調味料の成分分析を行った。可溶性総窒素(TN)は燃焼式(改良デュマ法)窒素測定装置デュマサーム(ゲルハルトジャパン社製)を用いて定法に従って測定した。Gly−Tyr、Ser−Tyr量は、LC−MS/MSシステム2695−QuattroMicro API(Waters社製)を用い、添加検量線法により求めた。結果を表2に示す。 The components of the obtained peptide-containing fermented seasoning were analyzed. Soluble total nitrogen (TN) was measured according to a conventional method using a combustion type (improved Dumas method) nitrogen measuring device Dumasarm (manufactured by Gerhard Japan Co., Ltd.). The amounts of Gly-Tyr and Ser-Tyr were determined by the addition calibration curve method using an LC-MS / MS system 2695-Quattro Micro API (manufactured by Waters). The results are shown in Table 2.
表2に示されるように、β−コングリシニン高含有大豆を使用した実施例1の発酵調味料は、一般的な国産大豆Aを用いた比較例1の発酵調味料よりも、降圧ペプチドであるGly−Tyr及びSer−Tyr量が顕著に高かった。 As shown in Table 2, the fermented seasoning of Example 1 using β-conglycinin-rich soybean is a hypotensive peptide Gly, as compared with the fermented seasoning of Comparative Example 1 using general domestic soybean A. The amounts of -Tyr and Ser-Tyr were significantly higher.
本発明は醤油又は醤油含有調味料などの製造分野において利用できる。 The present invention can be used in the field of manufacturing such as soy sauce or seasonings containing soy sauce.
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