JP7032619B2 - An antibacterial lactic acid strain that suppresses the growth of Bacillus bacteria, and a method for producing food using the strain. - Google Patents
An antibacterial lactic acid strain that suppresses the growth of Bacillus bacteria, and a method for producing food using the strain. Download PDFInfo
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Description
本発明は、バチルス属細菌の増殖を抑制できるなど、優れた抗菌活性をもつ乳酸菌、及び、その乳酸菌の添加によるバチルス属細菌の混入と増殖を抑えた食品の製造方法に関する。 The present invention relates to a lactic acid bacterium having excellent antibacterial activity such as being able to suppress the growth of Bacillus bacteria, and a method for producing a food product in which the contamination and growth of the Bacillus bacterium due to the addition of the lactic acid bacterium are suppressed.
味噌や醤油などの発酵調味料の製造現場では、しばしば、バチルス（Ｂａｃｉｌｌｕｓ）属細菌の混入による風味や見た目の変調（変敗）の影響により製品の歩留まりが大きく低下する問題を抱えている。 At the manufacturing site of fermented seasonings such as miso and soy sauce, there is often a problem that the yield of products is greatly reduced due to the influence of flavor and appearance alteration (deterioration) due to the contamination of Bacillus bacteria.
従来、バチルス属細菌の汚染や増殖による害を防ぎ、品質良好な発酵調味料を得るための発酵調味料の製造法としては、例えば、発酵調味料とするための原料を蒸煮し、製麹した後、麹を食塩水と仕込み、発酵させ、発酵調味料を得るに際し、前記蒸煮した原料を乳酸発酵して乳酸発酵物とし、この乳酸発酵物を製麹して麹とし、この麹を仕込む（特許文献１参照）ものが提案されている。 Conventionally, as a method for producing a fermented seasoning to prevent contamination and growth of Bacillus bacteria and to obtain a fermented seasoning with good quality, for example, the raw material for making the fermented seasoning is steamed and kneaded. Later, when the koji is charged with salt solution and fermented to obtain a fermented seasoning, the steamed raw material is lactic acid fermented to obtain a lactic acid fermented product, and this lactic acid fermented product is made into koji to be made into koji, and this koji is charged ( (See Patent Document 1) has been proposed.
また、上記の先行技術を発展させたものとして、大豆抽出液に浸漬した米を蒸煮して製造した蒸し米に、ラクトコッカス ラクティスを接種し、乳酸発酵させた後、製麹することで得た米麹と、蒸煮した大豆に、ペディオコッカス アシディラクティシを接種することで乳酸発酵した大豆を利用した、米味噌の製造方法（特許文献２参照）が提案されている。 In addition, as a development of the above-mentioned advanced technology, it was obtained by injecting lactococcus lactis into steamed rice produced by steaming rice soaked in soybean extract, fermenting it with lactic acid, and then making it into jiuqu. A method for producing rice miso (see Patent Document 2) has been proposed using soybeans lactic acid fermented by inoculating rice jiuqu and steamed soybeans with Pediococcus acidilactisi.
さらに、乳酸菌の抗菌作用を食品の製造工程に活かすものとしては、２００９年に日本でも食品添加物として認可されたナイシンが挙げられる。これは広い抗菌作用と安全性、風味に影響を及ぼさない点に特徴をもつ成分だが、麹菌が生産するプロテアーゼ（たんぱく質分解酵素）によって分解されることと、コストが割高である点が課題として残っている。 Furthermore, as a substance that utilizes the antibacterial action of lactic acid bacteria in the food manufacturing process, nisin, which was approved as a food additive in Japan in 2009, can be mentioned. This is a component characterized by its wide antibacterial action, safety, and no effect on flavor, but it remains a problem that it is decomposed by the protease (proteolytic enzyme) produced by Jiuqu and the cost is high. ing.
そして、特許文献２に発明の構成要件として記載されている乳酸菌について、ラクトコッカス ラクティスは、バクテリオシンの一種のナイシンを生産する乳酸菌であり、ペディオコッカス アシディラクティシもバクテリオシン生産菌である。以上の先行技術は、基本的にバクテリオシンの抗菌性を利用するものである。また、この抗菌性を利用する方法では、バチルス属細菌の汚染や増殖を防ぐと共に、味噌の風味も問題ないとしている。 Regarding the lactic acid bacterium described as a constituent requirement of the invention in Patent Document 2, Lactococcus lactis is a lactic acid bacterium that produces a kind of bacteriocin, nisin, and Pediococcus acidilactis is also a bacteriocin-producing bacterium. .. The above prior art basically utilizes the antibacterial property of bacteriocin. In addition, this method utilizing antibacterial properties prevents contamination and proliferation of Bacillus bacteria, and also states that there is no problem with the flavor of miso.
バチルス属細菌の増殖を抑制する抗菌性乳酸菌株、及び、それを用いた食品の製造方法に関して解決しようとする問題点は、従来のナイシンの抗菌性を利用する方法では、そのナイシンが、例えば麹菌が増殖する環境ではその麹菌が生産するプロテアーゼによって分解されるなど、その抗菌作用を効率的に発揮できない点と、食品の風味やうま味を積極的に向上させるものではない点にある。 The problem to be solved regarding the antibacterial lactic acid strain that suppresses the growth of Bacillus bacteria and the method of producing food using it is that in the conventional method utilizing the antibacterial property of nisin, the nisin is, for example, aspergillus. In an environment where lactic acid bacteria grow, it is decomposed by the protease produced by the aspergillus, and its antibacterial action cannot be exerted efficiently, and it does not positively improve the flavor and taste of food.
そこで本発明の目的は、バチルス属細菌の増殖を抑制できるなど、優れた抗菌活性をもつ乳酸菌、食品の風味やうま味を向上できる乳酸菌、及び、それを用いた食品の製造方法を提供することにある。 Therefore, an object of the present invention is to provide a lactic acid bacterium having excellent antibacterial activity such as suppressing the growth of Bacillus bacteria, a lactic acid bacterium capable of improving the flavor and umami of a food, and a method for producing a food using the lactic acid bacterium. be.
本発明は、上記目的を達成するために次の構成を備える。
本発明にかかる乳酸菌の一形態によれば、ペディオコッカス・ペントサセウスに属する乳酸菌株であって、米麹を原料として用いる食品を製造する場合において、前記米麹を得るために精米を原料として製麹を行う際に麹菌と共に添加されて前記麹菌と共に増殖させて使用できるもので、バチルス属細菌の増殖を抑制する抗菌性をもつ受託番号：ＮＩＴＥ Ｐ－０２５４３号として寄託されている乳酸菌ＰＰ１６５株であることを特徴とする。
The present invention comprises the following configurations in order to achieve the above object.
According to one form of the lactic acid bacterium according to the present invention, it is a lactic acid bacterium strain belonging to Pediococcus pentosaceus, and in the case of producing a food using rice jiuqu as a raw material, it is produced using milled rice as a raw material in order to obtain the rice jiuqu. It is a lactic acid bacterium PP165 strain that is added together with Jiuqu and can be used by growing it together with Jiuqu, and has antibacterial activity that suppresses the growth of Bacillus bacteria. It is characterized by being.
本発明にかかる抗菌性乳酸菌を用いた食品の製造方法の一形態によれば、前記の乳酸菌である乳酸菌ＰＰ１６５株を、精米を原料として製麹を行う際に麹菌と共に添加することで、前記麹菌と共に増殖させて米麹を得ることを特徴とすることができる。 According to one embodiment of the method for producing a food product using an antibacterial lactic acid bacterium according to the present invention, the lactic acid bacterium PP165 strain, which is the lactic acid bacterium, is added together with the aspergillus when the rice is milled as a raw material to produce the aspergillus. It can be characterized by growing with rice jiuqu to obtain rice jiuqu .
本発明にかかる抗菌性乳酸菌を用いた食品の製造方法の一形態によれば、前記の抗菌性乳酸菌を用いた食品の製造方法によって得られた米麹を、原料として用いて発酵調味料を製造することを特徴とすることができる。 According to one embodiment of the method for producing a food using an antibacterial lactic acid bacterium according to the present invention, a fermented seasoning is produced using the rice jiuqu obtained by the above-mentioned method for producing a food using an antibacterial lactic acid bacterium as a raw material. Can be characterized by doing.
本発明にかかる抗菌性乳酸菌を用いた食品の製造方法の一形態によれば、前記の抗菌性乳酸菌を用いた食品の製造方法によって得られた米麹を、原料として用いて味噌を製造することを特徴とすることができる。 According to one embodiment of the method for producing a food using an antibacterial lactic acid bacterium according to the present invention, miso is produced by using the rice jiuqu obtained by the above-mentioned method for producing a food using an antibacterial lactic acid bacterium as a raw material. Can be characterized.
本発明にかかる抗菌性乳酸菌を用いた食品の製造方法の一形態によれば、前記の抗菌性乳酸菌を用いた食品の製造方法によって得られた米麹を、原料として用いて酒や甘酒を製造することを特徴とすることができる。 According to one embodiment of the method for producing a food using an antibacterial lactic acid bacterium according to the present invention, sake or amazake is produced using the rice jiuqu obtained by the above-mentioned method for producing a food using an antibacterial lactic acid bacterium as a raw material. Can be characterized by doing.
本発明にかかるバチルス属細菌の増殖を抑制する抗菌性乳酸菌株、及び、それを用いた食品の製造方法によれば、高品質商品の製造に抗菌作用を効率的に発揮できることや、食品の風味やうま味を向上できるという特別有利な効果を奏する。 According to the antibacterial lactic acid bacterium strain that suppresses the growth of Bacillus bacteria according to the present invention and the method for producing foods using the same, the antibacterial action can be efficiently exhibited in the production of high-quality products and the flavor of foods. It has a special advantageous effect of improving the taste of umami.
以下、本発明にかかるバチルス属細菌の増殖を抑制する抗菌性乳酸菌株、及び、それを用いた食品の製造方法の形態例を、添付図面（図１、２）に基づいて詳細に説明する。 Hereinafter, a morphological example of an antibacterial lactic acid bacterium strain that suppresses the growth of Bacillus bacteria according to the present invention and a method for producing a food product using the same will be described in detail with reference to the accompanying drawings (FIGS. 1 and 2).
本発明では、味噌及び醤油もろみ素材中に存在する乳酸菌の抗菌作用に着目し、その味噌及び醤油もろみ素材から、変敗の主要原因菌として知られるバチルス・サブティリス（Ｂａｃｉｌｌｕｓ Ｓｕｂｔｉｌｉｓ）やバチルス・メチロトロフィカス（Ｂａｃｉｌｌｕｓ ｍｅｔｈｙｌｏｔｒｏｐｈｉｃｕｓ）を含むバチルス属細菌に対して抗菌活性をもつ複数の乳酸菌株の取得に成功した。１６ｓｒＤＮＡ領域の遺伝子配列解析による菌種判定の結果、これらの抗菌性菌株は、ペディオコッカス・ペントサセウス（Ｐｅｄｉｏｃｏｃｃｕｓ ｐｅｎｔｏｓａｃｅｕｓ）の菌株であることが明らかになった。
そして、そのペディオコッカス・ペントサセウスの複数の乳酸菌株のうち、受託番号：ＮＩＴＥ Ｐ－０２５４３号として寄託されている乳酸菌ＰＰ１６５株が、バチルス属細菌に対して最も強い抗菌活性を有することを確認した。
In the present invention, attention is paid to the antibacterial action of lactic acid bacteria existing in miso and soy sauce mash material, and from the miso and soy sauce mash material, Bacillus Subtilis and Bacillus methi, which are known as the main causative bacteria of deterioration. We have succeeded in obtaining a plurality of lactic acid bacteria strains having antibacterial activity against Bacillus bacteria including Bacillus methylotrophicus. As a result of strain determination by gene sequence analysis of the 16sr DNA region, it was clarified that these antibacterial strains are strains of Pediococcus pentosaceus.
Then, it was confirmed that among the plurality of lactic acid bacteria strains of Pediococcus pentosaceus, the lactic acid bacterium PP165 strain deposited under the accession number: NITE P-02543 has the strongest antibacterial activity against Bacillus bacteria. ..
このペディオコッカス・ペントサセウスを、食品の製造方法に用いる抗菌性乳酸菌として利用すれば、良質の食品を提供できる。すなわち、食品原料に、抗菌性乳酸菌としてのペディオコッカス・ペントサセウスを添加し、バチルス属細菌の生育を抑制することで、発酵調味料などの食品を高品質に製造することができる。
また、この抗菌活性をもつペディオコッカス・ペントサセウスは、バイオプリザバティブとして利用することが可能であり、食品の保存性を向上させる添加剤としても利用できる。
If this Pediococcus pentosaceus is used as an antibacterial lactic acid bacterium used in a method for producing food, good quality food can be provided. That is, by adding Pediococcus pentosaceus as an antibacterial lactic acid bacterium to a food raw material and suppressing the growth of Bacillus bacteria, it is possible to produce foods such as fermented seasonings with high quality.
In addition, Pediococcus pentosaceus having this antibacterial activity can be used as a biopreservative and can also be used as an additive for improving the preservation of foods.
実証試験によれば、抗菌性乳酸菌としてのペディオコッカス・ペントサセウスが、極めて優れた抗菌作用を有していることが確認された。なお、以下に示したペディオコッカス・ペントサセウスの実証試験結果は、乳酸菌ＰＰ１６５株のものである。例えば、このペディオコッカス・ペントサセウスを、製品（味噌、醤油豆）の製麹工程で、原料１ｇあたり１×１０７ＣＦＵ／ｍＬ以上添加し、同時にバチルス・スブティリスとバチルス・メチロトロフィカス（各１×１０３ＣＦＵ／ｍＬ）を添加する条件において、そのバチルス属２種は検出限界（１×１０２ＣＦＵ／ｍＬ）以下まで抑制されることが、実証試験によって確認された。なお、ペディオコッカス・ペントサセウスは、製麹工程で、生菌数が１×１０８ＣＦＵ／ｍＬ以上まで増殖し、食塩を添加した後の熟成期間において、塩分濃度が１２％の場合には、生菌数が１ヵ月後には１×１０３ＣＦＵ／ｍＬ程度となって２ヵ月後には１×１０２ＣＦＵ／ｍＬ程度まで減少したが、その抗菌作用は維持された。 According to the verification test, it was confirmed that Pediococcus pentosaceus as an antibacterial lactic acid bacterium has an extremely excellent antibacterial action. The verification test results of Pediococcus pentosaceus shown below are for the lactic acid bacterium PP165 strain. For example, this pediococcus pentosaceus is added at least 1 × 107 CFU / mL per 1 g of raw material in the koji making process of products (miso, soybeans), and at the same time Bacillus subtilis and Bacillus methirotropicus (1 each). It was confirmed by the verification test that the two Bacillus species were suppressed to the detection limit (1 × 10 2 CFU / mL) or less under the condition of adding × 10 3 CFU / mL). In addition, Pediococcus pentosaceus grows to a viable cell count of 1 × 108 CFU / mL or more in the koji making process, and when the salt concentration is 12% in the aging period after adding salt, the salt concentration is 12%. The viable cell count decreased to about 1 × 10 3 CFU / mL after 1 month and decreased to about 1 × 10 2 CFU / mL after 2 months, but its antibacterial activity was maintained.
また、醤油豆については、ペディオコッカス・ペントサセウスと上記バチルス属２種（各１×１０３ＣＦＵ／ｍＬ）を製麹工程で添加し、引き続く発酵熟成工程で、塩分濃度が５％の減塩条件下で実証試験を行ったが、ペディオコッカス・ペントサセウスが製麹工程で添加された試験区では、バチルス属２種を検出限界（１×１０２ＣＦＵ／ｍＬ）以下まで抑制できたのに対し、ペディオコッカス・ペントサセウスが添加されなかった試験区では、バチルス属２種が増殖した。これによって、ペディオコッカス・ペントサセウスはバチルス属２種に対する抗菌効果が優れていることが確認された。なお、この塩分濃度が５％の場合には、ペディオコッカス・ペントサセウスの生菌数が出麹時には１×１０９ＣＦＵ／ｍＬ程度となって熟成期間の１ヵ月後にも１×１０７ＣＦＵ／ｍＬ以上程度である状態を維持していた。 For soy sauce beans, Pediococcus pentosaceus and the above two species of Bacillus (1 x 10 3 CFU / mL each) are added in the koji making process, and the salt concentration is reduced by 5% in the subsequent fermentation and aging process. A verification test was conducted under the conditions, but in the test plot where Pediococcus pentosaceus was added in the fermentation process, two species of the genus Bacillus could be suppressed to the detection limit ( 1 × 102 CFU / mL) or less. On the other hand, in the test plot where Pediococcus pentosaceus was not added, two species of the genus Bacillus proliferated. From this, it was confirmed that Pediococcus pentosaceus has an excellent antibacterial effect against two species of the genus Bacillus. When this salt concentration is 5%, the viable cell count of Pediococcus pentosaceus is about 1 × 10 9 CFU / mL at the time of jiuqu, and 1 × 10 7 CFU / 1 month after the aging period. It maintained a state of about mL or more.
そして、本発明にかかる抗菌性乳酸菌（ペディオコッカス・ペントサセウス）は、タンパク質分解酵素に耐性を示し、高温でも抗菌活性が失活しない乳酸菌であり、生菌の状態で素材中に添加することで速やかに増殖し、バチルス属細菌に特に優れた抗菌作用を発揮する。増殖可能な塩分濃度は、５％程度以下であるが、製麹工程で添加することにより味噌や醤油などの発酵調味料の製造においても、その効果を充分に発揮させることができる。 The antibacterial lactic acid bacterium (Pediococcus pentosaceus) according to the present invention is a lactic acid bacterium that is resistant to proteolytic enzymes and does not inactivate its antibacterial activity even at high temperatures. It proliferates rapidly and exerts a particularly excellent antibacterial effect against Bacillus bacteria. The concentration of salt that can be grown is about 5% or less, but by adding it in the koji making process, the effect can be fully exerted even in the production of fermented seasonings such as miso and soy sauce.
ところで、抗菌作用を持つ乳酸菌を素材中に添加する食品の製造方法については、技術分野の欄で説明したように、従来の基準としても、素材中で良好に抗菌作用が発揮されることの他に、製品風味に悪影響を及ぼさない点が重要となっているが、風味やうま味を積極的に向上させる効果を期待するものではなかった。 By the way, as for the method of manufacturing a food in which lactic acid bacteria having an antibacterial action are added to a material, as explained in the section of the technical field, in addition to the fact that the antibacterial action is satisfactorily exhibited in the material even as a conventional standard. In addition, it is important that the product flavor is not adversely affected, but the effect of positively improving the flavor and umami was not expected.
これに対して、本発明にかかるペディオコッカス・ペントサセウスを利用した製品では、バチルス属細菌の増殖を検出限界以下まで抑制する効果に加え、塩慣れ効果（塩味を和らげる効果）など、従来の製造法と比較して製品風味の改善効果がみられている。 On the other hand, in the product using Pediococcus pentosaceus according to the present invention, in addition to the effect of suppressing the growth of Bacillus bacteria to the detection limit or less, the conventional production such as salt acclimatization effect (effect of softening salt taste) and the like. Compared with the method, the effect of improving the product flavor is seen.
有機酸の分析結果では、ペディオコッカス・ペントサセウスの作用後に、製品中にコハク酸、乳酸、酢酸などの旨味成分の増加が認められるため、これらの有機酸の複合的な作用により抗菌作用と風味改善作用が発揮されていることが考えられる。このような旨味成分の生産は、この菌株独自の特性によるもので、従来の乳酸菌株に対する優位性に相当すると考えられる。 According to the analysis results of organic acids, after the action of Pediococcus pentosaceus, an increase in succinic acid, lactic acid, acetic acid and other delicious components is observed in the product. It is considered that the flavor improving effect is exhibited. The production of such umami components is due to the unique characteristics of this strain, and is considered to correspond to the superiority over the conventional lactic acid strain.
味噌と醤油豆を製造する乳酸菌実証試験では、クエン酸とリンゴ酸の含有量は同等であったが、コハク酸、乳酸及び酢酸については、対照区に比べ、乳酸菌添加区で明らかに増加傾向がみられた。このように、有機酸が増加することで、バチルス菌に対する抗菌作用が強化されると共に、風味と旨味が向上していると考えられる。 In the lactic acid bacteria verification test for producing miso and soy sauce beans, the contents of citric acid and malic acid were the same, but succinic acid, lactic acid and acetic acid clearly increased in the lactic acid bacteria-added group compared to the control group. Was seen. As described above, it is considered that the increase in the amount of organic acid enhances the antibacterial action against Bacillus bacteria and improves the flavor and umami.
また、乳酸菌の特性解析によれば、ペディオコッカス・ペントサセウスの示す抗菌作用については、高温でも抗菌活性が失活しない乳酸菌であることが確認されていると共に、麹菌のたんぱく質分解酵素であるプロテアーゼの他に、プロティナーゼＫ、カタラーゼ、リパーゼによっても影響を受けないことが確認されている。 In addition, according to the characteristic analysis of lactic acid bacteria, it has been confirmed that the antibacterial action of Pediococcus pentosaceus is a lactic acid bacterium whose antibacterial activity is not inactivated even at high temperatures, and that the protease, which is a proteolytic enzyme of Jiuqu, is used. In addition, it has been confirmed that it is not affected by proteinase K, catalase, or lipase.
ペディオコッカス・ペントサセウスの耐熱性についての試験結果によれば、１００℃で６０分加熱後も、バチルス菌に対する抗菌作用に関する相対活性値が９０％以上であり、高温でも抗菌活性が失活しない乳酸菌であることを示した。この抗菌作用は、味噌製造の工程に想定されている温度帯だけでなく、ある程度の加熱を想定した他の食品製造の環境においても有効に作用することを示している。 According to the test results on the heat resistance of Pediococcus pentosaceus, the relative activity value for the antibacterial action against Bacillus is 90% or more even after heating at 100 ° C. for 60 minutes, and the antibacterial activity is not inactivated even at high temperature. It was shown that. It is shown that this antibacterial action is effective not only in the temperature range assumed in the process of producing miso but also in the environment of other food manufacturing assuming a certain degree of heating.
また、ペディオコッカス・ペントサセウスの酵素耐性についての試験結果によれば、バチルス菌２種に対する該乳酸菌の培養上清は、麹菌のプロテアーゼ、カタラーゼ、プロティナーゼＫ、リパーゼ処理に、相対活性値が９０％以上の強い抗菌作用を示し、酵素処理によっても影響されない高い酵素耐性を示した。従って、このペディオコッカス・ペントサセウスの抗菌作用は、プロテアーゼを生産する麹菌との共存下でも、十分に発揮されるものである。 In addition, according to the test results on the enzyme resistance of Pediococcus pentosaceus, the culture supernatant of the lactic acid bacterium against the two Bacillus bacteria has a relative activity value of 90% for the protease, catalase, proteinase K, and lipase treatment of the aspergillus. It showed the above-mentioned strong antibacterial action and showed high enzyme resistance that was not affected by enzyme treatment. Therefore, the antibacterial action of Pediococcus pentosaceus is sufficiently exerted even in the coexistence with the protease-producing aspergillus.
以上に説明したように、ペディオコッカス・ペントサセウスは、熱耐性の抗菌力及び各種の分解酵素耐性に優れ、麹菌と栄養分の奪い合いをすることがなく共存できて、麹菌と同時に増殖できる相性のよい抗菌性乳酸菌であって、しかも、麹菌の生長を阻害しないが、バチルス菌の増殖を高度に抑制できる抗菌性乳酸菌である。 As explained above, Pediococcus pentosaceus has excellent heat-resistant antibacterial activity and resistance to various degrading enzymes, can coexist with Jiuqu without competing for nutrients, and has good compatibility with Jiuqu and can grow at the same time. It is an antibacterial lactic acid bacterium, and it is an antibacterial lactic acid bacterium that does not inhibit the growth of aspergillus but can highly suppress the growth of Bacillus.
さらに、このペディオコッカス・ペントサセウスは、ＭＲＳ培地における生育について、塩分濃度５％及びアルコール濃度５％まで確認しており、幅広い多くの飲料品を含む食品製造に利用できる。すなわち、味噌や醤油の発酵調味料の他に、例えば、漬物などの発酵食品や酒又は甘酒などの発酵飲料品を含む食品の製造に広く利用できる。
また、このペディオコッカス・ペントサセウスであって、受託番号：ＮＩＴＥ Ｐ－０２５４３号として寄託されている乳酸菌ＰＰ１６５株は、その特性が優れており、食品（飲食料品）の製造に加え、動物用飼料や医薬品の製造における発酵を伴う工程においても、より広く適切に利用できる。
Furthermore, this Pediococcus pentosaceus has been confirmed to grow in MRS medium up to a salt concentration of 5% and an alcohol concentration of 5%, and can be used in the production of foods containing a wide variety of beverages. That is, it can be widely used in the production of foods including fermented foods such as pickles and fermented beverages such as liquor or amazake, in addition to fermented seasonings of miso and soy sauce.
In addition, the lactic acid bacterium PP165 strain deposited under the accession number: NITE P-02543 in this Pediococcus pentosaceus has excellent characteristics, and is used for animals in addition to the production of foods (food and drink). It can also be used more widely and appropriately in processes involving fermentation in the production of feeds and pharmaceuticals.
なお、抗菌性乳酸菌（ペディオコッカス・ペントサセウス）、及び、バチルス菌（バチルス・サブティリス）の生育と、塩分濃度との関係（塩分耐性）について、ＭＲＳ培地を用い、濁度測定法によって測定したところ、次の結果を得ている。先ず、塩分を添加しない場合は、抗菌性乳酸菌とバチルス菌のどちらも、急激に増殖する。また、塩分濃度が１０％では、抗菌性乳酸菌もバチルス菌も増殖が検出できないレベルで生育が抑制される。そして、塩分濃度が５％では、抗菌性乳酸菌とバチルス菌のどちらも、ゆっくりではあるが、増殖することが確認されている。なお、通常の味噌の塩分濃度は、一般的に１２％程度となっており、抗菌性乳酸菌とバチルス菌のどちらも、その生育が適正に抑制される。 The relationship between the growth of antibacterial lactic acid bacteria (Pediococcus pentosaceus) and Bacillus (Bacillus subtilis) and salinity (salt resistance) was measured using an MRS medium by a turbidity measurement method. However, the following results have been obtained. First, when no salt is added, both antibacterial lactic acid bacteria and Bacillus bacteria grow rapidly. Further, when the salt concentration is 10%, the growth of both antibacterial lactic acid bacteria and Bacillus bacteria is suppressed at a level at which growth cannot be detected. It has been confirmed that both antibacterial lactic acid bacteria and Bacillus bacteria grow slowly at a salt concentration of 5%. The salt concentration of ordinary miso is generally about 12%, and the growth of both antibacterial lactic acid bacteria and Bacillus bacteria is appropriately suppressed.
次に、本発明にかかるペディオコッカス・ペントサセウスを用いた発酵調味料の製造について説明する。
図１及び２に示す抗菌性乳酸菌を用いた食品（発酵調味料）の製造方法の工程例では、食品原料が、発酵調味料を製造するために加熱殺菌された発酵調味料用原料であって、その発酵調味料用原料に対するペディオコッカス・ペントサセウスの添加が、麹菌を添加することで行われる製麹工程で行われ、該製麹工程において前記麹菌と前記ペディオコッカス・ペントサセウスの両方を増殖させた後に、食塩を添加して発酵調味料を製造することを特徴としている。
Next, the production of a fermented seasoning using Pediococcus pentosaceus according to the present invention will be described.
In the process example of the method for producing a food (fermented seasoning) using antibacterial lactic acid bacteria shown in FIGS. 1 and 2, the food raw material is a raw material for fermented seasoning that has been heat-sterilized to produce the fermented seasoning. , The addition of pediococcus pentoseus to the fermented seasoning raw material is carried out in the koji making step performed by adding the aspergillus, and in the koji making step, both the aspergillus and the pediococcus pentoseus are propagated. It is characterized in that a fermented seasoning is produced by adding salt to the fermented seasoning.
発酵調味料のうちの味噌を製造する際の製造工程（図１参照）では、精米した米を水に浸漬して蒸煮した原料に、麹菌を添加する製麹工程において、ペディオコッカス・ペントサセウスを添加する。なお、製麹工程の期間は、例えば４８時間程度になっている。そして、その製麹工程が終わった原料に、大豆を水に浸漬して蒸煮した原料を混合し、食塩水を添加して諸味をつくり、その諸味を発酵熟成することで味噌を製造する。 In the manufacturing process for producing miso, which is one of the fermented seasonings (see Fig. 1), pediococcus pentosaceus is used in the koji making process in which the jiuqu is added to the raw material that is steamed by immersing the milled rice in water. Added. The period of the koji making process is, for example, about 48 hours. Then, the raw material obtained by immersing soybeans in water and steaming is mixed with the raw material for which the koji making process has been completed, and salt solution is added to make various flavors, and the various flavors are fermented and aged to produce miso.
発酵調味料のうちの醤油豆を製造する際の製造工程（図２参照）では、大豆を水に浸漬して蒸煮した原料に、麹菌を添加して製麹する製麹工程において、ペディオコッカス・ペントサセウスを添加する。その製麹工程が終わった原料に、調味料を加え発酵熟成することで醤油豆を製造する。
ところで、このペディオコッカス・ペントサセウスを原料に添加するタイミングは、麹菌の添加と同時でもよいし、麹菌の添加の後でもよく、厳格に限定されるものではなく、製麹工程において、麹菌とペディオコッカス・ペントサセウスのどちらも増殖できるように添加すればよい。
In the manufacturing process (see Fig. 2) when producing soybeans, which is one of the fermented seasonings, pediococcus is used in the manufacturing process of adding aspergillus to the raw material of soybeans soaked in water and steamed. -Add Pentoseus. Shoyumame is produced by adding seasonings to the raw materials for which the koji making process has been completed and fermenting and aging.
By the way, the timing of adding this Pediococcus pentosaceus to the raw material may be at the same time as the addition of the aspergillus or after the addition of the aspergillus, and is not strictly limited. Both Diococcus and Pentosaceus can be added so that they can grow.
ペディオコッカス・ペントサセウスは、抗菌性に優れているため、前述した塩分濃度が５％の醤油豆（醤油もろみ）の製造に関する実証試験結果にあるように、発酵調味料を極めて適切に減塩することが可能となる。なお、一般的に味噌などの発酵調味料の塩分濃度は１２％程度であり、通常の減塩された発酵調味料の塩分濃度は８％程度が限界であったが、本発明にかかるペディオコッカス・ペントサセウスを用いることで、塩分濃度が７％以下、さらには塩分濃度が５％以下の発酵調味料を適切に製造することが可能となる。 Since Pediococcus pentosaceus has excellent antibacterial properties, the fermented seasoning is extremely appropriately reduced as shown in the above-mentioned verification test results regarding the production of soy sauce beans (soy sauce mash) with a salt concentration of 5%. Is possible. In general, the salt concentration of fermented seasonings such as miso is about 12%, and the salt concentration of ordinary fermented seasonings with reduced salt is limited to about 8%. However, the pedio according to the present invention. By using Coccus pentosaceus, it becomes possible to appropriately produce a fermented seasoning having a salt concentration of 7% or less and further having a salt concentration of 5% or less.
さらに、液体培養されたペディオコッカス・ペントサセウスを、乾燥オカラの粉に混ぜて担持させることで菌担持粉状体とし、該菌担持粉状体を前記発酵調味料用原料に散布して添加することで、より均一に接種することができ、原料をムラなくより均一に発酵させることができる。 Further, the liquid-cultured Pediococcus pentosaceus is mixed with dried okara powder and supported to form a fungus-supported powder, and the fungus-supported powder is sprayed and added to the fermented seasoning raw material. As a result, the inoculum can be infused more uniformly, and the raw materials can be fermented more uniformly.
以上、本発明につき好適な形態例を挙げて種々説明してきたが、本発明はこの形態例に限定されるものではなく、発明の精神を逸脱しない範囲内で多くの改変を施し得るのは勿論のことである。 Although various examples of the present invention have been described above, the present invention is not limited to the examples of the present invention, and it goes without saying that many modifications can be made without departing from the spirit of the invention. That is.
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