JP6285068B1 - Nucleic acid-containing fermented seasoning and method for producing the same - Google Patents
Nucleic acid-containing fermented seasoning and method for producing the same Download PDFInfo
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- JP6285068B1 JP6285068B1 JP2017107512A JP2017107512A JP6285068B1 JP 6285068 B1 JP6285068 B1 JP 6285068B1 JP 2017107512 A JP2017107512 A JP 2017107512A JP 2017107512 A JP2017107512 A JP 2017107512A JP 6285068 B1 JP6285068 B1 JP 6285068B1
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- nucleic acid
- moromi
- fermented seasoning
- acid
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Abstract
【課題】５’−ヌクレオチド類を外部から添加することなく、発酵由来の５’−ヌクレオチド類のみで核酸を高濃度で含有する核酸含有醤油様調味料およびその製造方法を提供することを目的とする。【解決手段】大豆又は小麦等を主原料とした穀物原料に、麹菌を接種して固体麹を調製し、水又は食塩水を加えて食塩濃度４％（ｗ／ｖ）以下の諸味を加温分解する工程と、前記諸味を加熱処理してホスファターゼを失活させることにより殺菌諸味を調製する工程と、前記殺菌諸味に核酸生産微生物を接種して危害微生物の混入を抑制できる容器で核酸発酵を行う工程と、前記核酸生産微生物を自己消化して菌体内のＲＮＡを諸味中に遊離させることによりＲＮＡ抽出物を調製する工程と、前記ＲＮＡ抽出物にヌクレアーゼ及びデアミナーゼを作用させて呈味性のある５’−ヌクレオチド類に変換する工程と、を有する、発酵調味料の製造方法により解決する。【選択図】図１An object of the present invention is to provide a nucleic acid-containing soy sauce-like seasoning containing a high concentration of nucleic acids only from fermentation-derived 5'-nucleotides without adding 5'-nucleotides from the outside, and a method for producing the same. To do. [Solution] Grain raw materials mainly made from soybeans or wheat are inoculated with koji molds to prepare solid koji, and water or saline is added to warm the mash with a salt concentration of 4% (w / v) or less. The step of decomposing, the step of preparing the sterilized moromi by heat-treating the moromi to inactivate phosphatase, and inoculating the sterilized moromi with the nucleic acid-producing microorganism to inhibit the mixing of harmful microorganisms with nucleic acid fermentation A step of preparing an RNA extract by self-digesting the nucleic acid-producing microorganism and releasing the RNA in the microbial taste into moromi; and allowing the nuclease and deaminase to act on the RNA extract. It solves with the manufacturing method of a fermented seasoning which has the process of converting into a certain 5'-nucleotide. [Selection] Figure 1
Description
本発明は、製造工程中で核酸生産微生物により生産された核酸を高濃度で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 concentration of nucleic acid produced by a nucleic acid-producing microorganism during the production process and a method for producing the same.
醤油は「しょうゆ品質表示基準」によって「特級」「上級」「標準」に区別されている。これらの等級は「うま味」の指標といわれている「窒素分」の含量や色度（色の濃淡）などで定められており、一般的には、窒素分の含有量が多いほどうま味が豊かな醤油であるとされている。醤油のうま味に寄与する窒素分としては、グルタミン酸をはじめとしたアミノ酸類が知られている。 Soy sauce is classified into “special grade”, “advanced” and “standard” according to “soy sauce quality labeling standards”. These grades are determined by the content and chromaticity (color shade) of “nitrogen”, which is said to be an indicator of “umami”. Generally, the higher the nitrogen content, the richer the umami. It is said to be soy sauce. As nitrogen components contributing to the umami of soy sauce, amino acids such as glutamic acid are known.
一方、アミノ酸以外のうま味成分として、５’−ヌクレオチド類が知られている。５’−ヌクレオチド類とは、５’−アデニル酸、５’−グアニル酸、５’−イノシン酸及び５’−キサンチル酸等を始めとする呈味性ヌクレオチド類の総称であり、これらを含む調味料は核酸系調味料と呼ばれることもある。５’−ヌクレオチド類は単独でも強い呈味作用を有するが、醤油に含まれるＬ−グルタミン酸が存在すると、相乗効果により風味を顕著に改善強化し得ることが知られている。 On the other hand, 5'-nucleotides are known as umami components other than amino acids. 5′-nucleotides is a general term for taste nucleotides including 5′-adenylic acid, 5′-guanylic acid, 5′-inosinic acid, 5′-xanthylic acid, and the like. The ingredients are sometimes called nucleic acid seasonings. It is known that 5'-nucleotides alone have a strong taste effect, but when L-glutamic acid contained in soy sauce is present, the flavor can be significantly improved and enhanced by a synergistic effect.
しかしながら、醤油中には各種微生物に由来する多量のホスファターゼが存在しているため、醤油に呈味性ヌクレオチド類を添加しても、ヌクレオチド類はホスファターゼの作用により脱リン酸化されて呈味性のないヌクレオシドに分解される。 However, since soy sauce contains a large amount of phosphatase derived from various microorganisms, even if tasty nucleotides are added to soy sauce, the nucleotides are dephosphorylated by the action of phosphatase and have a tasty taste. Not broken down into nucleosides.
例えば、火入れ前の生醤油はこのホスファターゼ活性が特に強いものの一つで、添加した呈味性ヌクレオチド類は添加後１日でほとんど分解消失し、これらのヌクレオチド類の呈味性は全く消失してしまうことが知られている（特許文献１）。また、ホスファターゼ活性は、生醤油にかなり強力に存在するが、火入れ（８０℃達温）した醤油にも、生醤油の１０〜２５％程度残存していることが知られている。 For example, raw soy sauce before burning is one of the particularly strong phosphatase activities, and the added taste nucleotides almost completely decomposed and disappeared one day after the addition, and the taste of these nucleotides disappeared completely. (Patent Document 1). Moreover, although phosphatase activity exists quite strongly in raw soy sauce, it is known that about 10 to 25% of raw soy sauce remains in soy sauce that has been fired (temperature reached 80 ° C.).
したがって、ホスファターゼ活性を失活させることが、醤油に５’−ヌクレオチド類を添加し、安定に保持するためには必要であるとされている。従来知られている５’−ヌクレオチド類含有醤油の製造法は、常法により製造された醤油のホスファターゼ活性を失活させた後に５’−ヌクレオチド類を別途添加する方法であった（特許文献２、３）。しかしながら、醤油に核酸を添加した場合は、食品表示関連法規の規定に基づき、原材料に添加物の表示が必須となるため、「調味料（核酸）」と表示しなければならない。このような醤油様調味料は「化学調味料無添加」や、「天然醸造」を謳うことができず、天然志向、自然志向の消費者に対しては訴求力が弱かった。 Therefore, it is said that inactivating phosphatase activity is necessary in order to add 5'-nucleotides to soy sauce and keep it stable. A conventionally known method for producing 5′-nucleotide-containing soy sauce is a method in which 5′-nucleotides are added separately after inactivating the phosphatase activity of soy sauce produced by a conventional method (Patent Document 2). 3). However, when nucleic acid is added to soy sauce, the labeling of the additive is essential for the raw material based on the provisions of the food labeling laws and regulations, so “seasoning (nucleic acid)” must be displayed. Such a soy sauce-like seasoning cannot be used for “no chemical seasoning added” or “natural brewing”, and has weak appeal to natural-oriented and nature-oriented consumers.
呈味性ヌクレオチド類を発酵生産する公知の微生物として、例えば、高核酸型酵母エキスの製造に使用されているキャンディダ・ユティリス（Candida utilis）や核酸調味料の製造に用いられているコリネバクテリウム・グルタミカム（Corynebacterium glutamicum）等が知られている。仮にホスファターゼ活性の無い醤油もしくは諸味中でこれらの微生物を生育させ、核酸を発酵生産しようとしても、耐塩性が無いため、醤油や醤油諸味中では増殖することができず、核酸を高含有させることができない。このような背景から、従来の醤油製造業界では、５’−ヌクレオチド類を添加することなく、醤油諸味や圧搾後の生醤油、あるいは火入れ後の醤油中で５’−ヌクレオチド類を発酵によって生成蓄積することは極めて困難と考えられており、これまで実用化されていない。 Examples of known microorganisms that fermentatively produce tasty nucleotides include, for example, Candida utilis, which is used in the production of high nucleic acid yeast extract, and Corynebacterium, which is used in the production of nucleic acid seasonings. -Glutamicum (Corynebacterium glutamicum) etc. are known. Even if you try to grow these microorganisms in soy sauce or moromi with no phosphatase activity and try to fermentatively produce nucleic acids, they cannot grow in soy sauce or soy mash with high salt content. I can't. Against this background, in the conventional soy sauce manufacturing industry, 5′-nucleotides are produced and accumulated by fermentation in soy sauce moromi, pressed soy sauce, or soy sauce after burning without adding 5′-nucleotides. It is considered extremely difficult to do so and has not been put into practical use so far.
一方、醤油の醸造工程中での直接的な発酵生産ではないものの、呈味成分として核酸を含有する醤油の製造方法は幾つかの文献に記載されている。例えば、特許文献４には、天然醸造醤油の諸味の製造工程において、出麹した原料に加える仕込み水として、清酒等の発酵粕に加水し５０〜６０℃で１〜２週間自己消化させたもの、又はこれを圧搾した呈味性の高い液を用いることを特徴とする濃厚醤油の製造法が開示されている。しかし、特殊な原料（すなわち酒粕や焼酎蒸留廃液）を用いる必要がある。したがって、通常の醸造醤油と比較すると、風味もかなりかけ離れたものとなる欠点を有する。 On the other hand, although it is not direct fermentation production in the brewing process of soy sauce, a method for producing soy sauce containing nucleic acid as a taste component is described in several documents. For example, in Patent Document 4, in the production process of moromi of natural brewed soy sauce, water added to fermented rice cake such as sake as self-digested at 50 to 60 ° C. for 1 to 2 weeks as feed water to be added to the starting material. Alternatively, a method for producing a concentrated soy sauce characterized by using a highly tasteable liquid obtained by pressing this is disclosed. However, it is necessary to use special raw materials (ie, sake lees and shochu distillation waste liquid). Therefore, there is a drawback that the flavor is considerably different from that of ordinary brewed soy sauce.
本発明は、５’−ヌクレオチド類を外部から添加することなく、発酵由来の５’−ヌクレオチド類を高濃度に含有する発酵調味料およびその製造方法を提供することを目的とする。 The object of the present invention is to provide a fermented seasoning containing a high concentration of 5'-nucleotides derived from fermentation without adding 5'-nucleotides from outside, and a method for producing the same.
本発明者らは、上記課題を解決するために鋭意検討を行った結果、大豆又は小麦等を主原料とする穀物原料に麹菌を接種して調製した固体麹に、無塩又は低塩の仕込み水を加えて諸味を調製し、この諸味を加熱殺菌（ホスファターゼ失活）後、危害微生物の混入を抑制できる容器で汚染を防止しながら核酸発酵を行うことにより、耐塩性を有しない核酸発酵生産微生物による核酸発酵が可能となり、核酸を別途添加することなく、発酵由来の核酸を高濃度で含有する発酵調味料が得られることを見出し、本発明を完成させた。 As a result of intensive studies to solve the above-mentioned problems, the present inventors have prepared a solid koji prepared by inoculating koji mold on a cereal raw material mainly made of soybean or wheat, and charged with salt-free or low salt. Nucleic acid fermentation production without salt tolerance by preparing moromi by adding water, sterilizing this moromi with heat (phosphatase inactivation), and performing nucleic acid fermentation while preventing contamination in a container that can prevent the introduction of harmful microorganisms The present inventors have found that a fermented seasoning containing nucleic acid derived from fermentation at a high concentration can be obtained without adding nucleic acid separately, and nucleic acid fermentation by microorganisms is possible.
すなわち、本発明は、核酸（ただし添加由来の核酸を除く）１０ｐｐｍ（ｗ／ｖ）以上と、ＨＤＭＦ２ｐｐｍ（ｗ／ｖ）以上と、を含有する、発酵調味料を提供するものである。 That is, the present invention provides a fermented seasoning containing 10 ppm (w / v) or more of nucleic acid (excluding nucleic acids derived from addition) and 2 ppm (w / v) or more of HDMF.
また、本発明は、大豆又は小麦等を主原料とした穀物原料に、麹菌を接種して固体麹を調製し、水又は食塩水を加えて食塩濃度４％（ｗ／ｖ）以下の諸味を加温分解する工程と、前記諸味を加熱処理してホスファターゼを失活させることにより殺菌諸味を調製する工程と、前記殺菌諸味に核酸生産微生物を接種して危害微生物の混入を抑制できる容器で核酸発酵を行う工程と、前記核酸生産微生物を自己消化して菌体内のＲＮＡを諸味中に遊離させることによりＲＮＡ抽出物を調製する工程と、前記ＲＮＡ抽出物にリボヌクレアーゼ及びアデニル酸デアミナーゼを作用させて呈味性のある５’−ヌクレオチド類に変換する工程と、を有する、発酵調味料の製造方法を提供するものである。 In addition, the present invention prepares solid koji by inoculating koji molds on cereal raw materials mainly made from soybeans or wheat, and then adds water or saline to give a mash of 4% (w / v) or less in salt concentration. Nucleic acid in a container capable of incubating nucleic acid-producing microorganisms by inoculating nucleic acid-producing microorganisms into the sterilized moromi, and preparing a sterilized moromi by heating the moromi to inactivate phosphatase A step of fermenting, a step of preparing an RNA extract by self-digesting the nucleic acid-producing microorganism and releasing RNA in the microbial cells into moromi, and a reaction of ribonuclease and adenylate deaminase with the RNA extract. There is provided a method for producing a fermented seasoning having a step of converting to 5′-nucleotides having taste.
本発明によれば、５’−ヌクレオチド類（核酸）を外部から添加することなく、核酸生産微生物が発酵中に生産する５’−ヌクレオチド類のみで核酸を高濃度に含有する発酵調味料およびその製造方法を提供することができる。 According to the present invention, a fermented seasoning containing only 5′-nucleotides produced by a nucleic acid-producing microorganism during fermentation without adding 5′-nucleotides (nucleic acid) from the outside and containing the nucleic acid in a high concentration A manufacturing method can be provided.
以下、本発明に係る発酵調味料及びその製造方法を詳細に説明する。図１は、本実施形態に係る核酸含有発酵調味料の製造方法の工程図である。 Hereinafter, the fermented seasoning and the method for producing the same according to the present invention will be described in detail. FIG. 1 is a process diagram of a method for producing a nucleic acid-containing fermented seasoning according to this embodiment.
本発明において、核酸とは、５’−アデニル酸、５’−グアニル酸、５’−イノシン酸及び５’−キサンチル酸等を始めとする呈味性ヌクレオチド類を意味する。 In the present invention, the nucleic acid means tasty nucleotides including 5'-adenylic acid, 5'-guanylic acid, 5'-inosinic acid, 5'-xanthylic acid and the like.
本発明の実施形態においては、まず穀物原料より調製した固体麹に水又は食塩水を混和し、食塩濃度４％（ｗ／ｖ）以下、好ましくは食塩濃度４％（ｗ／ｖ）未満、より好ましくは食塩濃度０％（ｗ／ｖ）の醤油諸味を調製し、２５〜５７℃で０〜４８時間加温分解する。好ましくは、特許第３８２７３００号記載のように、７０〜８０℃の熱水又は食塩水を固体麹と混和し、５０〜５７℃に諸味温度を保持したまま、タンク内で間欠的又は連続的に撹拌し、１５〜３０時間酵素分解することが望ましい。 In an embodiment of the present invention, water or saline is first mixed in a solid koji prepared from a cereal raw material, and the salt concentration is 4% (w / v) or less, preferably less than 4% (w / v). Preferably, soy sauce moromi with a sodium chloride concentration of 0% (w / v) is prepared and thermally decomposed at 25-57 ° C. for 0-48 hours. Preferably, as described in Japanese Patent No. 3827300, hot water or saline at 70 to 80 ° C. is mixed with the solid koji and intermittently or continuously in the tank while maintaining the moromi temperature at 50 to 57 ° C. It is desirable to stir and enzymatically decompose for 15-30 hours.
ここで、穀物原料とは、例えば丸大豆、脱脂大豆、大豆タンパク、小麦グルテン、えんどう豆、そら豆、小豆等に代表される蛋白質原料と、小麦、大麦、ライ麦、フスマ、米、米ぬか、とうもろこし、澱粉粕等に代表される澱粉質原料を指す。これらは単独で、または組み合わせて用いることができる。 Here, the grain raw materials are protein raw materials such as whole soybeans, defatted soybeans, soybean protein, wheat gluten, peas beans, broad beans, red beans, etc., wheat, barley, rye, bran, rice, rice bran, corn , Refers to starchy raw materials represented by starch candy and the like. These can be used alone or in combination.
ここで用いられる麹（固体麹）は、常法により原料処理された蛋白質原料又はこれに澱粉質原料を混合したものに、アスペルギルス・ソーヤ（Aspergillus sojae）、アスペルギルス・オリゼ（Aspergillus oryzae）に代表される麹菌を接種し、２〜３日固体培養（製麹）することで得られる。蛋白質原料に澱粉質原料を混合する場合、配合割合は特に限定するものではないが、例えば通常の醤油に近い調味料を得ようとする場合、重量比で１：０．２５〜４とすることが好ましい。 The koji (solid koji) used here is typified by a protein raw material processed by a conventional method or a mixture of a starch raw material and a protein raw material, such as Aspergillus sojae and Aspergillus oryzae. It is obtained by inoculating a koji mold and solid-culturing (making a koji) for 2 to 3 days. When the starch raw material is mixed with the protein raw material, the blending ratio is not particularly limited. For example, when trying to obtain a seasoning close to normal soy sauce, the weight ratio should be 1: 0.25-4. Is preferred.
仕込みに用いる水又は食塩水は、麹が十分浸る程度であればよく、一般には、麹重量に対し１〜１０容量倍（ｖ／ｗ）とすることが好ましい。また、加温分解時に、防ばい性や分解効率の向上、風味向上のために後述する食用の酸や酵素剤、活性炭を添加してもよい。 The water or the salt solution used for preparation should just be a grade which a soot can fully immerse, and generally it is preferable to set it as 1-10 volume times (v / w) with respect to the soot weight. Moreover, you may add the edible acid, enzyme agent, and activated carbon which are mentioned later for the improvement of antibacterial property, decomposition | disassembly efficiency, and flavor improvement at the time of a thermal decomposition.
加温分解時には、諸味の分解促進のため、酵素剤を終濃度が０．００１〜１％（ｗ／ｖ）となるように添加してもよい。酵素剤としては、例えば、プロテアーゼ（エンドプロテアーゼ、エキソプロテアーゼ）、セルラーゼ、ペクチナーゼ等を挙げることができる。 At the time of thermal decomposition, an enzyme agent may be added so that the final concentration is 0.001 to 1% (w / v) in order to promote the decomposition of moromi. Examples of the enzyme agent include protease (endoprotease, exoprotease), cellulase, pectinase and the like.
次に加温分解を行った食塩濃度４％（ｗ／ｖ）以下の固液分離前の醤油諸味について加熱処理を行ない、殺菌諸味を調製する。この加熱処理により、上述の製麹工程において麹菌が生産したホスファターゼ等の核酸分解酵素を失活させると共に、諸味の殺菌を行う。ここで、殺菌機の負荷軽減や、核酸生産微生物の生育速度向上を目的として、殺菌前に適宜加水を行って諸味を希釈してもよい。 Next, heat treatment is performed on the soy sauce moromi before solid-liquid separation with a salt concentration of 4% (w / v) or less, which has been subjected to thermal decomposition, to prepare sterilized moromi. This heat treatment inactivates nucleolytic enzymes such as phosphatase produced by koji molds in the above-described koji making process and sterilizes moromi. Here, for the purpose of reducing the load on the sterilizer and improving the growth rate of the nucleic acid-producing microorganism, the moromi may be diluted by appropriate addition of water before sterilization.
ここで加熱処理の方法は特に限定されず、ＵＨＴ、ＨＴＳＴ、レトルト、加圧タンク、スチームインジェクション、スチームインフュージョン、オートクレーブ、プレートヒーター、表面かきとり式、ジュール式熱交換、チューブラー式熱交換等の加熱処理方法のいずれを用いてもよいが、好ましくは加圧タンクやチューブラー式熱交換機を用いるのがよい。例えば、諸味を加圧タンクに入れ、均一に撹拌しながら加圧加温することなどで核酸分解酵素の失活や殺菌を行うことができる。加熱温度が低すぎる、もしくは加熱時間が短すぎると、核酸分解酵素の失活や雑菌の殺菌が不十分となるため好ましくない。逆に、加熱温度が高すぎる、もしくは加熱時間が長すぎると、調味料の風味が劣化するために好ましくない。選択する加熱方法により最適条件は異なるが、例えば、８０℃では２分以上１８０分以下、１２１℃では５秒以上１５分以下、１３０℃では１秒以上３０秒以下が好ましい。 Here, the heat treatment method is not particularly limited, and UHT, HTST, retort, pressurized tank, steam injection, steam infusion, autoclave, plate heater, scraped surface, Joule heat exchange, tubular heat exchange, etc. Any of the heat treatment methods may be used, but a pressurized tank or a tubular heat exchanger is preferably used. For example, the nucleolytic enzyme can be deactivated or sterilized by putting moromi in a pressurized tank and heating under pressure while stirring uniformly. If the heating temperature is too low or the heating time is too short, the nuclease deactivation and sterilization of various bacteria become insufficient, which is not preferable. Conversely, if the heating temperature is too high or the heating time is too long, the flavor of the seasoning is deteriorated, which is not preferable. Optimum conditions vary depending on the heating method to be selected. For example, it is preferably 2 minutes to 180 minutes at 80 ° C., 5 seconds to 15 minutes at 121 ° C., and 1 second to 30 seconds at 130 ° C.
調製した諸味は、防ばい性の向上や味の調整のために、ｐＨ調整を行ってもよい。調整後のｐＨは、防ばい性と酵母の発酵性の観点から、３．０〜７．０、好ましくは４．０〜５．５にすることが望ましい。ｐＨ調整のタイミングとしては、加温分解時、諸味の加熱処理前、諸味の加熱処理後のいずれでもよい。ｐＨ調整剤としての食用の酸としては、例えば、乳酸、酢酸、リンゴ酸、クエン酸、グルコン酸、アジピン酸等が挙げられるが、風味の点から、乳酸が好ましい。 The prepared moromi may be adjusted for pH in order to improve anti-bacterial properties and adjust the taste. The pH after the adjustment is desirably 3.0 to 7.0, preferably 4.0 to 5.5, from the viewpoints of antifungal properties and yeast fermentability. The timing of pH adjustment may be any of the time of thermal decomposition, before the heat treatment of moromi, or after the heat treatment of moromi. Examples of the edible acid as the pH adjuster include lactic acid, acetic acid, malic acid, citric acid, gluconic acid, adipic acid and the like, and lactic acid is preferred from the viewpoint of flavor.
諸味には核酸生産微生物による核酸発酵を円滑に行うため、糖質を０〜２０％（ｗ／ｖ）添加してもよい。例えば、グルコース、フルクトース、スクロース、マルトース、マンノース、グリセロール等、核酸生産微生物が資化できる糖質が利用できるが、資化効率を考えるとグルコースの使用が望ましい。また、これらの糖を含む食品素材、例えば、砂糖、ブドウ糖果糖液糖、果糖ブドウ糖液糖、三温糖、糖蜜などを用いてもよい。 In order to smoothly carry out the nucleic acid fermentation by the nucleic acid-producing microorganism, 0 to 20% (w / v) of saccharides may be added to the moromi. For example, carbohydrates that can be assimilated by nucleic acid-producing microorganisms, such as glucose, fructose, sucrose, maltose, mannose, glycerol, etc. can be used, but considering the efficiency of utilization, the use of glucose is desirable. In addition, food materials containing these sugars, such as sugar, glucose fructose liquid sugar, fructose glucose liquid sugar, trivalent sugar, molasses, etc. may be used.
また、核酸発酵の前にも、諸味の分解促進や圧搾性の向上のため、酵素剤を終濃度が０．００１〜１％（ｗ／ｖ）となるように添加してもよい。酵素剤としては、例えば、プロテアーゼ（エンドプロテアーゼ、エキソプロテアーゼ）、セルラーゼ、ペクチナーゼ等を挙げることができる。 Moreover, before nucleic acid fermentation, you may add an enzyme agent so that final concentration may be 0.001-1% (w / v) in order to accelerate | stimulate decomposition | disassembly of moromi and to improve pressability. Examples of the enzyme agent include protease (endoprotease, exoprotease), cellulase, pectinase and the like.
また、核酸発酵の前に、苦味を除去して風味を向上させるため、諸味に活性炭を添加してもよい。活性炭は粉末であることが好ましく、平均粒子径が１０〜１００μｍのものを使用することがより好ましい。活性炭の添加量は、諸味原料に対し０．１〜５％（ｗ／ｗ）であることが好ましい。活性炭の種類は用途に応じて適宜選択することができ、例えば、苦味除去、悪臭除去、味の調整、色の調整またはこれらの機能を有する活性炭を組み合わせて使用することができる。 In addition, activated carbon may be added to moromi to remove bitterness and improve flavor before nucleic acid fermentation. The activated carbon is preferably a powder, and more preferably one having an average particle diameter of 10 to 100 μm. The addition amount of the activated carbon is preferably 0.1 to 5% (w / w) with respect to the moromi material. The type of the activated carbon can be appropriately selected depending on the application, and for example, bitterness removal, malodor removal, taste adjustment, color adjustment, or a combination of activated carbons having these functions can be used.
また、核酸発酵の前に、前記殺菌諸味を固液分離してもよい。固液分離の方法は特に限定されず、例えば、圧搾、遠心分離、ろ過等の方法で実施することができる。 In addition, the sterilization moromi may be solid-liquid separated before nucleic acid fermentation. The method of solid-liquid separation is not particularly limited, and for example, the solid-liquid separation can be performed by a method such as pressing, centrifugation, or filtration.
本発明で使用する核酸生産微生物は、核酸を生産することができる微生物であれば特に制限されず、例えばキャンディダ・ユティリス（Candida utilis）、サッカロマイセス・セレビシエ（Saccharomyces cerevisiae）、ジゴサッカロマイセス・ルーキシー（Zygosaccharomyces rouxii）、シゾサッカロマイセス・ポムベ（Schizosaccharomyces pombe）、クリュイベロマイセス・マルシアヌス（Kluyveromyces marxianus）などの酵母、コリネバクテリウム・グルタミカム（Corynebacterium glutamicum）、コリネバクテリウム・アンモニアゲネス（Corynebacterium ammoniagenes）、バチルス・サブチリス（Bacillus subtilis）などの細菌、またはそれらの栄養要求・アナログ耐性変異株が用いられる。本発明の諸味は無塩又は低塩であるがゆえに、本来、耐塩性の弱い酵母や細菌を使用することができる。 The nucleic acid-producing microorganism used in the present invention is not particularly limited as long as it is a microorganism capable of producing a nucleic acid. For example, Candida utilis, Saccharomyces cerevisiae, Zygosaccharomyces rouxii), yeasts such as Schizosaccharomyces pombe, Kluyveromyces marxianus, Corynebacterium glutamicum, Corynebacterium ammoniagenes, Bacillus ammoniagenes Bacteria such as Bacillus subtilis, or auxotrophic / analog resistant mutants thereof are used. Since the moromi of the present invention is salt-free or low-salt, yeast or bacteria with low salt tolerance can be used.
これら核酸生産微生物の添加濃度は、酵母であれば、諸味１ｇあたり１×１０４個以上、好ましくは１×１０５〜１×１０７個となるように添加するのが好ましく、細菌であれば、諸味１ｇあたり１×１０４個以上、好ましくは１×１０５〜１×１０７個となるように添加するのが好ましい。 The addition concentration of these nucleic acid-producing microorganisms is 1 × 10 4 or more, preferably 1 × 10 5 to 1 × 10 7 per 1 g of moromi in the case of yeast. It is preferable to add 1 × 10 4 or more, preferably 1 × 10 5 to 1 × 10 7 moromi.
諸味は危害微生物の混入を抑制できる容器に投入され、この容器内で核酸発酵を行う。ここで、危害微生物の混入を抑制できる容器とは、容器内部と外気を遮断できる構造を持つものであればよく、実験的にはポリプロピレン製の滅菌済み広口瓶やガラス製のメディアボトルなどが使用でき、工業的には容器内に除菌された空気を供給できる機能を持つジャーファーメンターや加圧式の発酵タンクなどを使用することができる。また、空気の除菌には、０．３μｍ以上の塵を９９．９７％以上集塵できるフィルター、例えばＨＥＰＡフィルターなどを用いることができる。 The moromi is put into a container capable of suppressing the contamination of harmful microorganisms, and nucleic acid fermentation is performed in this container. Here, the container that can prevent the introduction of harmful microorganisms may be any container that has a structure that can block the inside of the container and the outside air. Experimentally, a sterilized wide-mouth bottle made of polypropylene or a glass media bottle is used. Industrially, a jar fermenter having a function of supplying sterilized air into the container, a pressurized fermentation tank, or the like can be used. For sterilization of air, a filter that can collect 99.97% or more of dust of 0.3 μm or more, such as a HEPA filter, can be used.
核酸発酵は、核酸生産微生物が生育できる温度、具体的には１５〜４５℃、好ましくは２０〜３５℃で１〜９０日、好ましくは２〜２８日行う。 Nucleic acid fermentation is performed at a temperature at which a nucleic acid-producing microorganism can grow, specifically 15 to 45 ° C, preferably 20 to 35 ° C, for 1 to 90 days, preferably 2 to 28 days.
本実施形態に係る発酵調味料に含まれるアルコールは、核酸発酵工程で添加する糖濃度や使用する微生物の種類や温度条件によって０〜２０％（ｗ／ｖ）と産生量が調整でき、また、核酸発酵終了時にアルコールを添加してもよいが、発酵調味料として良好な風味を持たせるために、好ましくは８％（ｗ／ｖ）未満、より好ましくは２〜７％（ｗ／ｖ）とするのがよい。 The alcohol contained in the fermented seasoning according to the present embodiment can be adjusted in production amount from 0 to 20% (w / v) depending on the sugar concentration added in the nucleic acid fermentation process, the type of microorganism used and the temperature condition, Alcohol may be added at the end of nucleic acid fermentation, but in order to have a good flavor as a fermented seasoning, it is preferably less than 8% (w / v), more preferably 2-7% (w / v). It is good to do.
核酸発酵の終了後は、諸味中の核酸生産微生物を自己消化させ、もしくはプロテアーゼやβ−グルカナーゼを含む酵素剤を用いて酵母細胞壁を溶解させ、菌体内のＲＮＡを諸味中に遊離させる。自己消化は諸味を一定時間加温することにより行うことができる。自己消化は例えば核酸生産微生物のＲＮＡの溶出が最大となるのに十分な条件、例えば品温４０〜６０℃で１分間〜２４時間行うことがより好ましい。品温が４０℃未満では自己消化に時間を要し、また諸味液汁に対する核酸生産微生物由来のＲＮＡの溶出が十分でなく、反対に６０℃を越える温度では諸味の加熱劣化による褐変や風味の悪化を招くので好ましくない。時間が１分間未満では核酸生産微生物由来のＲＮＡの溶出が十分でなく、反対に２４時間を越えると加熱劣化により諸味の褐変や風味の悪化を招くので好ましくない。また自己消化の際に核酸の遊離効率を向上させる目的で、消化の前に内在性の核酸分解酵素を加熱失活させる方法や、凍結融解を行う方法、エタノール等の有機溶媒を添加する方法、超音波処理を行う方法、ｐＨ４〜６程度の酸性もしくはｐＨ８〜１２程度のアルカリ性に調整する方法等、公知の方法を組み合わせて使用することもできる。また、酵母細胞壁を溶解する酵素剤としては、ツニカーゼ（商標、アマノエンザイム社製）、デナチームGEL（商標、ナガセケムテックス社製）、フィルトラーゼBRX（商標、DSM社製）、等を定法に従って使用することができる。 After completion of nucleic acid fermentation, nucleic acid-producing microorganisms in moromi are self-digested, or yeast cell walls are lysed using an enzyme agent containing protease and β-glucanase to release RNA in the microbial taste into the moromi. Autolysis can be performed by warming moromi for a certain period of time. The autolysis is more preferably performed under conditions sufficient for maximizing elution of RNA from the nucleic acid-producing microorganism, for example, at a product temperature of 40 to 60 ° C. for 1 minute to 24 hours. If the product temperature is less than 40 ° C, it takes time for self-digestion, and RNA from microorganisms producing nucleic acids is not sufficiently eluted in moromi soup. On the other hand, if the temperature exceeds 60 ° C, browning or deterioration of flavor due to aging of moromi This is not preferable. If the time is less than 1 minute, elution of RNA derived from the nucleic acid-producing microorganism is not sufficient. On the other hand, if it exceeds 24 hours, browning of taste and deterioration of flavor are caused by heat deterioration. Also, for the purpose of improving the efficiency of nucleic acid release during autolysis, a method of inactivating endogenous nucleolytic enzyme before digestion, a method of freezing and thawing, a method of adding an organic solvent such as ethanol, It is also possible to use a combination of known methods such as a method of performing ultrasonic treatment, a method of adjusting the acidity to about pH 4-6 or an alkalinity of about pH 8-12. In addition, as an enzyme agent that dissolves the yeast cell wall, Tunicase (trademark, manufactured by Amano Enzyme), Denateam GEL (trademark, manufactured by Nagase ChemteX), Filtolase BRX (trademark, manufactured by DSM), etc. are used in accordance with conventional methods. can do.
核酸生産微生物の自己消化又は酵素消化を行った後、呈味性のないＲＮＡ抽出物に酵素処理を行って呈味性のある５’−ヌクレオチド類（核酸）に変換する。酵素はヌクレアーゼを使用することができ、これに加えてデアミナーゼを使用することが好ましい。ヌクレアーゼとしてはリボヌクレアーゼ、デオキシリボヌクレアーゼ等が挙げられ、リボヌクレアーゼであることが好ましい。さらに、リボヌクレアーゼの中でも切断様式によりＰ１ヌクレアーゼ、Ｓ１ヌクレアーゼ等が挙げられ、いずれの酵素も使用することができる。また、デアミナーゼとしてはアデニル酸デアミナーゼであることが好ましい。これらの酵素はいずれも市販の各種酵素を使用することができる。または麦芽もしくは発芽米等、これらの酵素を含む食品を添加することもできる。酵素処理における温度、ｐＨ、添加量、処理時間等は、使用する酵素の種類、活性の強さ、所望の核酸濃度等を考慮して適宜決定することができる。また、酵素処理を実施する順序はヌクレアーゼ処理物に対してデアミナーゼ処理が可能であればよく、ヌクレアーゼ処理を実施した後のＲＮＡ抽出物にデアミナーゼを添加してデアミナーゼ処理を行ってもよく、用いるヌクレアーゼとデアミナーゼが共に同じ温度で酵素活性を示す場合には、ＲＮＡ抽出物にヌクレアーゼとデアミナーゼを共に添加して同時に酵素処理を行ってもよい。 After self-digestion or enzymatic digestion of the nucleic acid-producing microorganism, the RNA extract having no taste is subjected to an enzyme treatment to convert it into taste-preferred 5'-nucleotides (nucleic acids). As the enzyme, nuclease can be used, and in addition to this, deaminase is preferably used. Examples of the nuclease include ribonuclease and deoxyribonuclease, and ribonuclease is preferable. Furthermore, among ribonucleases, P1 nuclease, S1 nuclease and the like can be mentioned depending on the cleavage mode, and any enzyme can be used. The deaminase is preferably adenylate deaminase. Any of these enzymes can use various commercially available enzymes. Alternatively, foods containing these enzymes such as malt or germinated rice can be added. The temperature, pH, addition amount, treatment time and the like in the enzyme treatment can be appropriately determined in consideration of the type of enzyme used, the strength of activity, the desired nucleic acid concentration, and the like. The order of performing the enzyme treatment is not limited as long as the deaminase treatment can be performed on the nuclease-treated product, and the deaminase treatment may be performed by adding deaminase to the RNA extract after the nuclease treatment. And deaminase both exhibit enzymatic activity at the same temperature, the nuclease and deaminase may be added to the RNA extract and simultaneously subjected to enzymatic treatment.
例えば、不溶物が除去されたＲＮＡ抽出物をｐＨ４．０以上、ｐＨ７未満、好ましくはｐＨ４．５〜６．０に調整した後、ヌクレアーゼを添加して６０〜８０℃で３０分間〜１２時間ヌクレアーゼ処理した後、デアミナーゼを添加して４０〜６０℃で３０分間〜６時間デアミナーゼ処理を行うことにより、イノシン酸とグアニル酸を充分に生成することができる。 For example, after adjusting the RNA extract from which insolubles have been removed to pH 4.0 or more and less than pH 7, preferably pH 4.5 to 6.0, nuclease is added and nuclease is added at 60 to 80 ° C. for 30 minutes to 12 hours. After the treatment, inosinic acid and guanylic acid can be sufficiently generated by adding deaminase and performing a deaminase treatment at 40 to 60 ° C. for 30 minutes to 6 hours.
酵素処理工程が終了した諸味は、そのまま圧搾せずに諸味（味噌・ペースト）様調味料とすれば、発酵由来の核酸を高濃度に含む諸味様調味料とすることができる。また、諸味を圧搾、火入れ、清澄、濾過等の常法による処理を行うことで、発酵由来の核酸を高濃度に含む発酵調味料（例えば、醤油様調味料）を得ることができる。 If the moromi after the enzyme treatment process is used as a moromi (miso / paste) -like seasoning without pressing as it is, a moromi-like seasoning containing a high concentration of fermentation-derived nucleic acid can be obtained. Moreover, the fermented seasoning (for example, soy sauce-like seasoning) which contains the nucleic acid derived from fermentation in high concentration can be obtained by processing the moromi taste by conventional methods, such as pressing, burning, clarification, and filtration.
酵素処理工程が終了した後、諸味に食塩を添加することにより、食塩含有発酵調味料を調製することもできる。すなわち、諸味の核酸発酵は無塩又は低塩条件下で行うことで核酸発酵を促進させ、それにより得られた発酵諸味に任意の濃度の食塩を添加することで、醤油らしい風味を有する発酵調味料を製造することができる。食塩の添加量は適宜設定することができ、食塩濃度によって、減塩醤油様調味料や濃口醤油様調味料を容易に調製することができる。 After completion of the enzyme treatment step, a salt-containing fermented seasoning can be prepared by adding salt to the moromi. In other words, moromi nucleic acid fermentation is carried out under salt-free or low-salt conditions to promote nucleic acid fermentation, and by adding any concentration of salt to the fermentation moromi obtained thereby, a fermented seasoning that has a taste like soy sauce The material can be manufactured. The amount of salt added can be set as appropriate, and a reduced salt soy sauce-like seasoning or concentrated soy sauce-like seasoning can be easily prepared depending on the salt concentration.
本実施形態の発酵調味料は、添加由来の核酸を除く核酸濃度が１０ｐｐｍ以上である。核酸濃度は核酸発酵工程で添加する糖濃度や使用する微生物の種類や温度条件によって適宜調整できるが、核酸濃度は核酸生産微生物の核酸生産能に依存するため、上限値は自ずと核酸生産微生物の種類によって決まる。例えば、上述した核酸生産酵母で核酸発酵を行った場合は５０００ｐｐｍ程度が上限値となる。なお、核酸発酵後に加熱減圧濃縮、スプレードライ、凍結乾燥等の公知の方法によって濃縮加工を行い、発酵調味料中の核酸濃度を上記上限値以上に高めることも可能である。 In the fermented seasoning of this embodiment, the nucleic acid concentration excluding the nucleic acid derived from the addition is 10 ppm or more. The nucleic acid concentration can be adjusted as appropriate depending on the sugar concentration added in the nucleic acid fermentation process, the type of microorganism used, and the temperature conditions. However, since the nucleic acid concentration depends on the nucleic acid producing ability of the nucleic acid producing microorganism, the upper limit is naturally the type of nucleic acid producing microorganism. It depends on. For example, when nucleic acid fermentation is performed with the above-described nucleic acid-producing yeast, the upper limit is about 5000 ppm. In addition, it is also possible to raise the nucleic acid concentration in a fermented seasoning to the above upper limit value or more by performing a concentration process by a known method such as heating under reduced pressure, spray drying, freeze drying and the like after nucleic acid fermentation.
一般的なこいくち醤油の核酸含量が検出限界以下であるのに対して、本実施形態の発酵調味料は上記のように著量の核酸を含有している。そのため、醤油に元々含まれるグルタミン酸（０．２〜２．０％（ｗ／ｖ）程度）との相乗効果により、本実施形態に係る発酵調味料及びそれを用いた飲食品の旨味・コクが顕著に向上する。仮に核酸を外部から添加した場合は、原材料に添加物の表示が必須となるため、化学調味料無添加や、天然醸造を謳うことができないが、本実施形態の発酵調味料であれば原材料名に添加物の表示が不要なため、自然志向、天然指向の消費者に対し付加価値を訴求できる。 While the nucleic acid content of common koikuchi soy sauce is below the detection limit, the fermented seasoning of this embodiment contains a significant amount of nucleic acid as described above. Therefore, due to a synergistic effect with glutamic acid (about 0.2 to 2.0% (w / v)) originally contained in soy sauce, the fermented seasoning according to this embodiment and the umami and richness of food and drink using the same Remarkably improved. If nucleic acid is added from the outside, the labeling of the additive is essential for the raw material, so no chemical seasoning can be added or natural brewing cannot be obtained, but if it is a fermented seasoning of this embodiment, the name of the raw material Since no additive labeling is required, added value can be appealed to nature-oriented and nature-oriented consumers.
本実施形態に係る発酵調味料はまた、香気成分のひとつである4-ヒドロキシ-2,5-ジメチル-3(2H)-フラノン（ヒドロキシ・ジメチル・フラノン、ＨＤＭＦとも呼ばれる）濃度が２ｐｐｍ（ｗ／ｖ）以上であり、より好ましい風味を有している。ＨＤＭＦは代表的なフラノン化合物であり、砂糖様の甘さとフルーティーさとを併せ持ち、醤油等、呈味強化に寄与する調味料の代表的香気成分として知られている。本発明の発酵調味料は、一般のこいくち醤油と同等以上のＨＤＭＦを含有しており、醤油のような香ばしい風味に優れていることが特徴である。ＨＤＭＦ濃度が２ｐｐｍより低いと、醤油のような香ばしい風味が不十分となってしまう。本実施形態に係る発酵調味料に含まれるＨＤＭＦは、原料である麹から発酵を経て産生されたものであり、上述した核酸と同様、原材料名に添加物の表示が不要なため、自然志向、天然指向の消費者に対し付加価値を訴求できる。 The fermented seasoning according to the present embodiment also has a concentration of 2-ppm-2,5-dimethyl-3 (2H) -furanone (also called hydroxy-dimethyl furanone, HDMF), which is one of aroma components, of 2 ppm (w / v) It is above and has a more preferable flavor. HDMF is a typical furanone compound, which has both sugar-like sweetness and fruity, and is known as a typical aroma component of seasonings such as soy sauce that contributes to enhancing taste. The fermented seasoning of the present invention contains HDMF that is equal to or higher than ordinary soy sauce, and is characterized by an excellent savory flavor like soy sauce. If the HDMF concentration is lower than 2 ppm, the savory flavor like soy sauce will be insufficient. The HDMF contained in the fermented seasoning according to the present embodiment is produced through fermentation from the raw material koji, and, like the nucleic acid described above, it is not necessary to display the additive in the name of the raw material. Appeal value added to nature-oriented consumers.
本実施形態の他の実施形態として、通常の方法で調製した生醤油又は火入れ醤油を電気透析等で脱塩して食塩濃度を下げることにより食塩濃度４％（ｗ／ｖ）以下の脱塩生醤油又は脱塩火入れ醤油とし、その後は上述したように加熱殺菌（ホスファターゼ失活）、無菌的な核酸発酵、呈味性核酸への変換を実施してもよい。 As another embodiment of the present embodiment, fresh soy sauce or hot soy sauce prepared by a normal method is desalted by electrodialysis or the like to lower the salt concentration, thereby reducing the salt concentration to 4% (w / v) or less. Or you may use desalted fired soy sauce, and you may implement heat sterilization (phosphatase inactivation), aseptic nucleic acid fermentation, and conversion to a taste nucleic acid as mentioned above after that.
すなわち、通常の方法で調製した生醤油又は火入れ醤油を電気透析等で脱塩して食塩濃度を下げることにより食塩濃度４％（ｗ／ｖ）以下の脱塩生醤油又は脱塩火入れ醤油を調製する。次に、脱塩生醤油又は脱塩火入れ醤油をジャケット付きタンクで加熱したり、プレートヒーター又はチューブヒーター等の熱交換器で加熱処理することにより、脱塩生醤油又は脱塩火入れ醤油中のホスファターゼを失活させると共に脱塩生醤油又は脱塩火入れ醤油を殺菌する。そして、殺菌した脱塩生醤油又は脱塩火入れ醤油を危害微生物の混入が抑制できる容器に無菌的に移し、核酸生産微生物を植菌し、先に説明した発酵条件で核酸発酵を行う。その後核酸生産微生物を自己消化して菌体内のＲＮＡを諸味中に遊離させることによりＲＮＡ抽出物を調製し、ＲＮＡ抽出物にリボヌクレアーゼ等のヌクレアーゼとアデニル酸デアミナーゼ等のデアミナーゼを作用させれば、呈味性のある５’−ヌクレオチド類に変換され、所望の発酵調味液を製造することができる。 That is, fresh salted soy sauce or salted fired soy sauce with a salt concentration of 4% (w / v) or less is prepared by desalting raw soy sauce or fired soy sauce prepared by a normal method by electrodialysis or the like to lower the salt concentration. . Next, the desalted raw soy sauce or desalted fired soy sauce is heated in a jacketed tank or heat-treated with a heat exchanger such as a plate heater or tube heater to lose phosphatase in the desalted fresh soy sauce or desalted fired soy sauce. Sterilize fresh salted soy sauce or desalted fired soy sauce. Then, sterilized desalted raw soy sauce or desalted fired soy sauce is aseptically transferred to a container capable of suppressing contamination with harmful microorganisms, inoculated with nucleic acid-producing microorganisms, and subjected to nucleic acid fermentation under the fermentation conditions described above. After that, an RNA extract is prepared by self-digesting the nucleic acid-producing microorganism and releasing the RNA in the microbial taste into moromi. The desired fermented seasoning liquid can be produced by converting to 5′-nucleotides having taste.
本発明の発酵調味料は、そのままでも天然調味料や、日本農林規格で定義される醤油と同様の使い方ができ、また任意の飲食品に配合することもできる。そのため、本発明の発酵調味料を含有した、うま味やコクが改善した飲食品を提供することができる。飲食品の具体例としては、例えば、しょうゆ加工品、つゆ、たれ、ぽん酢、和風だし、洋風だし、中華だし、ドレッシング、スープ、ソース、惣菜のもと、等の調味料原料；野菜、果実、穀物等の加工品を含む農産加工食品；魚介類、海藻等の加工品を含む水産加工食品；卵・乳製品等の加工品を含む畜産加工食品；等を挙げることができる。本発明の発酵調味料を含有した飲食品は、最終製品の核酸含量が高まるため、うま味の向上やおいしさを高めることができる。その上、最終製品の原材料表示において、発酵調味料としての表示が可能で、核酸の添加物表示は不要となる利点を有する。 The fermented seasoning of the present invention can be used as it is as a natural seasoning or soy sauce defined in Japanese Agricultural Standards, or can be blended in any food or drink. Therefore, the food / beverage products which improved the umami | taste and richness containing the fermented seasoning of this invention can be provided. Specific examples of foods and drinks include, for example, soy sauce processed products, soy sauce, sauce, ponzu, Japanese-style soup, Western-style soup, Chinese soup stock, dressing, soup, sauce, source of side dish, etc .; vegetables, fruits, Agricultural processed foods including processed products such as cereals; marine processed foods including processed products such as seafood and seaweed; livestock processed foods including processed products such as eggs and dairy products. Since the food and drink containing the fermented seasoning of the present invention increases the nucleic acid content of the final product, it can improve the umami taste and taste. In addition, in the display of the raw material of the final product, it is possible to display it as a fermented seasoning, and there is an advantage that the display of the nucleic acid additive is unnecessary.
以下、実施例により本発明をさらに具体的に説明する。ただし、本発明の技術的範囲は、それらの例により何ら限定されるものではない。 Hereinafter, the present invention will be described more specifically with reference to examples. However, the technical scope of the present invention is not limited by these examples.
（実施例１）高核酸発酵調味料の調製
１．醤油麹の作製
脱脂加工大豆５０％（ｗ／ｗ）と焙煎割砕小麦５０％（ｗ／ｗ）の配合割合で、定法に従い醤油麹を作製した。なお、脱脂加工大豆は１３０％（ｗ／ｗ）撒水し蒸煮したものを用いた。この原料にアスペルギルス・ソーヤ（Aspergillus sojae）の種麹を接種し、２５℃〜４０℃、湿度９５％の環境で４２時間製麹して醤油麹を得た。
(Example 1) Preparation of high nucleic acid fermentation seasoning Production of soy sauce cake Soy sauce cake was produced according to a conventional method at a blending ratio of 50% (w / w) of defatted soybeans and 50% (w / w) of roasted cracked wheat. The defatted soybean used was 130% (w / w) watered and steamed. This raw material was inoculated with seeds of Aspergillus sojae and koji 42 hours in an environment of 25 ° C. to 40 ° C. and 95% humidity to obtain a soy sauce koji.
２．諸味の調製
上記醤油麹１００重量部に対し、２００重量部の７０℃に加温した熱水（食塩は含まない）を混和し、回転軸に撹拌翼を配置した保温ジャケット付きの分解タンク内で連続的に１００ｒｐｍで撹拌し、５５℃で２４時間加温分解を行い、無塩の諸味を得た。
2. Preparation of moromi In 100 parts by weight of the soy sauce cake, 200 parts by weight of hot water heated to 70 ° C. (without salt) is mixed, and in a decomposition tank with a heat insulation jacket with a stirring blade on the rotating shaft. The mixture was continuously stirred at 100 rpm and subjected to thermal decomposition at 55 ° C. for 24 hours to obtain a salt-free moromi.
３．諸味の殺菌およびホスファターゼ失活処理
上記無塩諸味６００ｇを２．５Ｌ容のジャーファーメンター（バイオット社製）に入れ、リン酸２水素カリウム（和光純薬工業社製）１２ｇ、消泡剤（信越シリコーン社製）０．６ｍＬ、水道水３６０ｍＬを加え、水酸化カリウム（和光純薬工業社製）を添加してｐＨ５．５となるよう調整し、オートクレーブで１２１℃、５分の殺菌を行った。
3. Moromi sterilization and phosphatase deactivation treatment 600 g of the above salt-free moromi is placed in a 2.5 L jar fermenter (Biot), 12 g of potassium dihydrogen phosphate (Wako Pure Chemical Industries, Ltd.), defoamer (Shin-Etsu) (Made by Silicone Co., Ltd.) 0.6 mL and tap water 360 mL were added, potassium hydroxide (manufactured by Wako Pure Chemical Industries, Ltd.) was added to adjust to pH 5.5, and sterilization was carried out at 121 ° C. for 5 minutes in an autoclave. .
４．核酸発酵
上記無塩諸味を３０℃に冷却し、核酸生産酵母としてキャンディダ・ユティリス（Candida utilis）を植菌し、定法に従って３０℃で４８時間の培養を行った。別途滅菌した５０％グルコース水溶液を１．７ｇ／ｈｒで流加し、無菌空気を１Ｌ／ｍｉｎ、内圧０．０４ＭＰａで通気した。撹拌速度は６００ｒｐｍとした。
4). Nucleic acid fermentation The salt-free moromi was cooled to 30 ° C., Candida utilis was inoculated as a nucleic acid-producing yeast, and cultured at 30 ° C. for 48 hours according to a conventional method. Separately sterilized 50% glucose aqueous solution was fed at 1.7 g / hr, and sterile air was aerated at 1 L / min and an internal pressure of 0.04 MPa. The stirring speed was 600 rpm.
５．酵素処理
培養終了後、ツニカーゼ（商標、アマノエンザイム社製）を３００ｍｇ添加し、５０℃で２時間撹拌した。続いて、ヌクレアーゼアマノＧ（商標、アマノエンザイム社製）を３００ｍｇ加え、６５℃で１時間撹拌した。さらに、デアミザイム（商標、アマノエンザイム社製）を３００ｍｇ加え、４５℃で１時間撹拌した。最後に８０℃で２０分間撹拌して酵素を失活させ、定法に従って濾紙（アドバンテック社製）で濾過を行い、核酸を高含有する発酵（醤油様）調味液を得た（サンプル１）。
5. Enzyme treatment After completion of the culture, 300 mg of tunicase (trademark, manufactured by Amano Enzyme) was added and stirred at 50 ° C. for 2 hours. Subsequently, 300 mg of nuclease Amano G (trademark, manufactured by Amano Enzyme) was added and stirred at 65 ° C. for 1 hour. Further, 300 mg of deamizyme (trademark, manufactured by Amano Enzyme) was added and stirred at 45 ° C. for 1 hour. Finally, the enzyme was inactivated by stirring at 80 ° C. for 20 minutes, followed by filtration with filter paper (manufactured by Advantech) according to a conventional method to obtain a fermented (soy sauce-like) seasoning liquid containing a high amount of nucleic acid (Sample 1).
６．発酵調味料の成分分析
（１）呈味性核酸の定量
呈味性核酸の定量は、既報（東京都立衛生研究所研究年報、p172-175、2001年）を改変した方法で、高速液体クロマトグラフィー（ＨＰＬＣ）を用いて行った。すなわち、固相抽出カラムSep-Pak Light QMA（ウォーターズ社製）を用い、下記の要領で前処理を行った。
↓メタノール 2 mL（活性化）
↓超純水 4 mL（平衡化）
↓Sample 300 uL
（アンモニア50 uLと混合し350 uLをロード）
↓超純水 3 mL（非吸着画分）
↓1%ギ酸 5 mL（吸着画分）
↓遠心濃縮装置で蒸発乾固
↓HPLCの溶離液Aに再溶解
6). Component analysis of fermented seasonings (1) Quantification of taste nucleic acids Nutrition of taste nucleic acids was determined by high performance liquid chromatography using a method modified from the previous report (Tokyo Metropolitan Institutes of Health Research Annual Report, p172-175, 2001). (HPLC). That is, pretreatment was performed using a solid phase extraction column Sep-Pak Light QMA (Waters) in the following manner.
↓ Methanol 2 mL (activated)
↓ Ultrapure water 4 mL (equilibrium)
↓ Sample 300 uL
(Mixed with 50 uL of ammonia and loaded with 350 uL)
↓ Ultrapure water 3 mL (non-adsorbed fraction)
↓ 1% formic acid 5 mL (adsorption fraction)
↓ Evaporate to dryness with centrifugal concentrator ↓ Re-dissolve in HPLC eluent A
次に、この前処理済みサンプルをＨＰＬＣ（島津製作所社製）で分析した。条件は下記のとおり。
カラム：Shiseido CAPCELL PAK C18 MGIII 4.6 x 250 mm（資生堂社製）
溶離液Ａ：0.1% トリエチルアミン + 1% メタノール／超純水、 pH 5.0（リン酸で調整）
溶離液Ｂ：80% アセトニトリル／超純水
流速：1.0 mL/min、 検出：UV 254 nm
Next, this pretreated sample was analyzed by HPLC (manufactured by Shimadzu Corporation). The conditions are as follows.
Column: Shiseido CAPCELL PAK C18 MGIII 4.6 x 250 mm (manufactured by Shiseido)
Eluent A: 0.1% triethylamine + 1% methanol / ultra pure water, pH 5.0 (adjusted with phosphoric acid)
Eluent B: 80% acetonitrile / ultra pure water Flow rate: 1.0 mL / min, detection: UV 254 nm
標準品として用いた５’−ＩＭＰおよび５’−ＧＭＰの検量線から、定量を行った。その結果、核酸濃度は表１のとおりであった。ただし、濃度は５．酵素処理で得た発酵調味液の液量あたりの濃度（ｗ／ｖ）である。 Quantification was performed from calibration curves of 5'-IMP and 5'-GMP used as standards. As a result, the nucleic acid concentration was as shown in Table 1. However, the concentration is 5. It is the density | concentration (w / v) per volume of the fermented seasoning liquid obtained by the enzyme treatment.
（２）ＨＤＭＦの定量
ＨＤＭＦの濃度は、Ｊ．Ａｇｒｉｃ．Ｆｏｏｄ Ｃｈｅｍ．Ｖｏｌ．３９，９３４，１９９１記載の定量分析法により実施した。より具体的には、ガスクロマトグラフィー（アジレント・テクノロジーズ社製６８９０Ｎ）による分析を行い、標準物質を用いた検量線法により、各種香気成分含有量を決定した。
(2) Quantification of HDMF Agric. Food Chem. Vol. The quantitative analysis method described in 39,934,1991 was carried out. More specifically, an analysis by gas chromatography (Agilent Technologies 6890N) was performed, and various aroma component contents were determined by a calibration curve method using a standard substance.
（実施例２）醤油様調味液に核酸を添加した効果
特許第５８３６４６６号公報の実施例２−１に記載の方法で調製した無塩醤油様調味料に、食塩濃度が４％（ｗ／ｖ）未満になるように食塩を添加し、さらに５’−ＩＭＰおよび５’−ＧＭＰを主成分として含有する調味料（商品名：リボタイド、ＭＣフードスペシャリティーズ社製、５’−ＩＭＰ：５’−ＧＭＰ含有量比＝１：１）を各濃度で添加して、所望の醤油様調味液を調製した（サンプル２〜７）。これらの各サンプルについて、以下の要領で官能評価を行った。なお、対照の減塩醤油に元々含まれる核酸量は検出限界以下であることを分析により確認した。官能評価は、識別能力を有するパネル7名がサンプルの原液をスポイトで直接口に含み、核酸を添加していない対照の醤油様調味液と味に違いがあるかどうか評価し、違いを識別できた人数で示した。さらに、パネル全員の討議により、総合的な味の好ましさを記号で示した（表２）。なお、記号は以下の評価を表す。◎：対照と比べて非常に好ましい、○：対照と比べて好ましい、×：対照と同等。
(Example 2) Effect of adding nucleic acid to soy sauce-like seasoning solution Salt-free soy sauce-like seasoning prepared by the method described in Example 2-1 of Japanese Patent No. 5836466 has a salt concentration of 4% (w / v). ) And a seasoning containing 5′-IMP and 5′-GMP as main components (trade name: Ribotide, manufactured by MC Food Specialties, Inc., 5′-IMP: 5′- GMP content ratio = 1: 1) was added at each concentration to prepare desired soy sauce-like seasonings (samples 2 to 7). Each of these samples was subjected to sensory evaluation in the following manner. It was confirmed by analysis that the amount of nucleic acid originally contained in the control low-salt soy sauce was below the detection limit. In sensory evaluation, seven panelists with discrimination ability can directly identify the difference by including the sample stock solution directly in the mouth with a dropper and evaluating whether there is a difference in taste from the control soy sauce-like seasoning solution to which no nucleic acid is added. Shown by the number of people. Furthermore, the overall taste of the taste was indicated by symbols after discussion by all the panels (Table 2). The symbol represents the following evaluation. (Double-circle): It is very preferable compared with a control | contrast, (circle): It is preferable compared with a control | contrast, x: It is equivalent to a control | contrast.
表２より、核酸濃度が１０ｐｐｍ以上のサンプル（サンプル３〜７）ではパネル全員が味の違いを識別し、特に１００ｐｐｍ以上では旨味や味の持続性が向上しているというコメントがあった。この結果から、本発明に類似の製造法によって得られる醤油様調味料に、１０ｐｐｍ以上の核酸を含有させることにより、醤油様調味料の味を改善できることが示された。ただし、核酸濃度が１００００ｐｐｍでは、核酸の味が強すぎ、醤油様調味料としての味のバランスが崩れるというパネルのコメントがあったため、総合評価としては対照と同程度となった。 According to Table 2, all the panels identified the difference in taste in samples (samples 3 to 7) having a nucleic acid concentration of 10 ppm or more, and there was a comment that umami and taste persistence were improved particularly at 100 ppm or more. From this result, it was shown that the taste of soy sauce-like seasoning can be improved by adding 10 ppm or more of nucleic acid to the soy sauce-like seasoning obtained by the production method similar to the present invention. However, when the nucleic acid concentration was 10,000 ppm, the panel comment that the taste of the nucleic acid was too strong and the taste balance as a soy sauce-like seasoning was lost, so the overall evaluation was comparable to the control.
（実施例３）醤油に核酸含有発酵調味料を添加した効果
減塩醤油（キッコーマン社製、市販品）に、本発明の実施例１で調製した核酸含有醤油様調味料（サンプル１）を配合量を変えて添加し、核酸濃度が１〜１０００ｐｐｍの醤油様調味液を調製した。これらの各サンプルについて、以下の要領で食塩濃度が９%（ｗ／ｖ）になるよう食塩を添加して、所望の醤油様調味料（サンプル８〜１２）を調製した。これらの各サンプルについて、実施例２と同様の方法で官能評価を行った。
(Example 3) Effect of adding nucleic acid-containing fermented seasoning to soy sauce Reduced salt soy sauce (manufactured by Kikkoman Co., Ltd., commercial product) is blended with nucleic acid-containing soy sauce-like seasoning (sample 1) prepared in Example 1 of the present invention. A soy sauce-like seasoning liquid having a nucleic acid concentration of 1-1000 ppm was prepared by changing the amount. About each of these samples, salt was added so that salt concentration might be 9% (w / v) in the following ways, and the desired soy sauce-like seasoning (samples 8-12) was prepared. Each of these samples was subjected to sensory evaluation in the same manner as in Example 2.
表３より、核酸濃度が１０ｐｐｍ以上になるように本発明の核酸含有発酵調味料を添加すると、減塩醤油の味が顕著に改善することが確認された。なお、核酸濃度が１０００ｐｐｍでは、核酸含有発酵調味料の配合量が比較的多かったため、対照の減塩醤油とはやや異なる味が感じられたが、全体としては品質が改善した。また、本発明では著量のＨＤＭＦを含有することから、醤油のような香ばしい香りを有し、発酵調味料として優れた品質を有することが確認された。 From Table 3, it was confirmed that when the nucleic acid-containing fermented seasoning of the present invention was added so that the nucleic acid concentration was 10 ppm or more, the taste of the reduced salt soy sauce was remarkably improved. When the nucleic acid concentration was 1000 ppm, the amount of the nucleic acid-containing fermented seasoning was relatively large, so a taste slightly different from the control low-salt soy sauce was felt, but the quality as a whole improved. Moreover, in this invention, since it contains a remarkable amount of HDMF, it has a fragrant fragrance like soy sauce and has been confirmed to have excellent quality as a fermented seasoning.
（実施例４）核酸分解酵素の失活条件
実施例１の無塩諸味４ｇに水道水６ｍＬを加え、これを５０、６０、７０、８０℃で１０分間、または１２１℃で２分間の条件で加熱した。続いて、各諸味に５’−グアニル酸９００ｐｐｍ、５’−イノシン酸９００ｐｐｍを添加し、３０℃で４６時間振とうした。その後、遠心分離によって固形分を除去し、上澄み液を０．４５μｍフィルター（アドバンテック社製）に通した。
(Example 4) Inactivation conditions of nucleolytic enzyme 6 mL of tap water was added to 4 g of the salt-free moromi of Example 1, and this was added at 50, 60, 70, 80 ° C for 10 minutes, or 121 ° C for 2 minutes. Heated. Subsequently, 900 ppm of 5′-guanylic acid and 900 ppm of 5′-inosinic acid were added to each moromi and shaken at 30 ° C. for 46 hours. Thereafter, the solid content was removed by centrifugation, and the supernatant was passed through a 0.45 μm filter (manufactured by Advantech).
呈味性核酸の定量は高速液体クロマトグラフ質量分析計（ＬＣ−ＭＳ）を用いて行った。条件は下記の通り。
カラム：Poroshell 120 PFP 3.0 x 150 mm（アジレント社製）
溶離液Ａ：0.1% ギ酸／超純水
溶離液Ｂ：0.1% ギ酸／アセトニトリル
流速：0.4 mL/min
イオン化：ESI（＋）
検出：SIMモード
The taste nucleic acid was quantified using a high performance liquid chromatograph mass spectrometer (LC-MS). The conditions are as follows.
Column: Poroshell 120 PFP 3.0 x 150 mm (manufactured by Agilent)
Eluent A: 0.1% formic acid / ultra pure water Eluent B: 0.1% formic acid / acetonitrile Flow rate: 0.4 mL / min
Ionization: ESI (+)
Detection: SIM mode
標準添加法に従い、５’−ＩＭＰ及び５’−ＧＭＰの標品を用いて定量を行った。その結果を図１に示す。図１は、無塩諸味を加熱した後に呈味性核酸（５’−グアニル酸及び５’−イノシン酸）を添加し、経時的に核酸濃度を定量した結果を示す図である。 According to the standard addition method, quantification was performed using 5'-IMP and 5'-GMP preparations. The result is shown in FIG. FIG. 1 is a diagram showing the results of quantifying the nucleic acid concentration over time by adding tasty nucleic acids (5'-guanylic acid and 5'-inosinic acid) after heating salt-free moromi.
図１に示すように、加熱温度が７０℃以下の場合は１８時間処理した時点で呈味性核酸が完全に分解されたが、加熱温度８０℃以上の場合は処理開始から４６時間まで呈味性核酸は分解されなかった。従って、核酸分解酵素を失活させるためには８０℃１０分間の滅菌が必要となることが判明した。 As shown in FIG. 1, when the heating temperature is 70 ° C. or lower, the tasty nucleic acid is completely decomposed at the time of treatment for 18 hours. Sex nucleic acid was not degraded. Therefore, it was found that sterilization at 80 ° C. for 10 minutes is necessary to inactivate the nucleolytic enzyme.
（実施例５）各種酵母による核酸発酵
実施例１の無塩諸味４０ｇに、リン酸２水素カリウム１ｇ、消泡剤０．１ｇ、水道水６０ｍＬを加え、水酸化カリウムでｐＨ５．５となるよう調整し、オートクレーブで１２１℃、２分の殺菌を行った。
(Example 5) Nucleic acid fermentation with various yeasts To 40 g of salt-free moromi of Example 1, 1 g of potassium dihydrogen phosphate, 0.1 g of antifoaming agent and 60 mL of tap water are added, and the pH is adjusted to 5.5 with potassium hydroxide. The mixture was sterilized by autoclaving at 121 ° C. for 2 minutes.
上記培地に別途滅菌した５０％グルコース溶液を５ｍＬ添加し、キャンディダ・ユティリス（Candida utilis）（サンプル１３）またはサッカロマイセス・セレビシエ（Saccharomyces cerevisiae）（サンプル１４）またはジゴサッカロマイセス・ルキシー（Zygosaccharomyces rouxii）（サンプル１５）またはクルイベロマイセス・マルキシアヌス（Kluyveromyces marxianus）（サンプル１６）を植菌し、定法に従って３０℃で２４時間の培養を行った。 5 mL of a 50% glucose solution sterilized separately is added to the above medium, and Candida utilis (sample 13) or Saccharomyces cerevisiae (sample 14) or Zygosaccharomyces rouxii (sample) 15) or Kluyveromyces marxianus (sample 16) was inoculated and cultured at 30 ° C. for 24 hours according to a conventional method.
培養終了後、培養液２０ｍＬから遠心分離で上澄みを除去し、得られた沈殿を水に懸濁して酵母懸濁液２０ｍＬを調製した。この酵母懸濁液にエタノール０．５ｍＬを添加した後、ｐＨを８に調整し、５０℃で２時間振とうした。続いて、ｐＨを５に調整し、ヌクレアーゼアマノＧ（商標、アマノエンザイム社製）を７．５ｍｇ加え、６５℃で１時間反応させた。さらに、デアミザイム（商標、アマノエンザイム社製）を７．５ｍｇ加え、５５℃で１時間反応させた。最後に８５℃で２０分間加温して酵素を失活させ、遠心分離によって固形分を取り除き、核酸発酵調味液（サンプル１３〜１６）を得た。 After completion of the culture, the supernatant was removed from the culture solution 20 mL by centrifugation, and the resulting precipitate was suspended in water to prepare a yeast suspension 20 mL. After adding 0.5 mL of ethanol to this yeast suspension, the pH was adjusted to 8, and the mixture was shaken at 50 ° C. for 2 hours. Subsequently, the pH was adjusted to 5, 7.5 mg of nuclease Amano G (trademark, manufactured by Amano Enzyme) was added, and the mixture was reacted at 65 ° C. for 1 hour. Furthermore, 7.5 mg of deamizyme (trademark, manufactured by Amano Enzyme) was added and reacted at 55 ° C. for 1 hour. Finally, the enzyme was inactivated by heating at 85 ° C. for 20 minutes, the solid content was removed by centrifugation, and nucleic acid fermentation seasonings (samples 13 to 16) were obtained.
実施例４と同様の条件で核酸発酵調味液を分析したところ、核酸濃度は表４のとおりであった。表４より、キャンディダ・ユティリス以外の酵母も核酸発酵調味液の製造に使用できることが示された。 When the nucleic acid fermentation seasoning liquid was analyzed under the same conditions as in Example 4, the nucleic acid concentrations were as shown in Table 4. From Table 4, it was shown that yeasts other than Candida utilis can also be used for the production of a nucleic acid fermentation seasoning liquid.
（実施例６）ＨＤＭＦ濃度の比較
本発明品と市販の高核酸酵母エキスに含まれるＨＤＭＦ濃度を比較するため、核酸発酵調味液を作製した。
(Example 6) Comparison of HDMF concentration In order to compare the HDMF concentration contained in the product of the present invention and a commercially available high nucleic acid yeast extract, a nucleic acid fermentation seasoning solution was prepared.
実施例１の無塩諸味５２０ｇに、リン酸２水素カリウム１３ｇ、消泡剤０．６ｇ、水道水３８０ｍＬを加え、水酸化カリウムでｐＨ５．５となるよう調整し、オートクレーブで１１５℃、１５分の殺菌を行った。この培地にキャンディダ・ユティリスを植菌し、定法に従って３０℃で２２時間の培養を行った。別途滅菌した５０％グルコース水溶液を８．４ｇ／ｈｒで流加し、無菌空気を１Ｌ／ｍｉｎ、内圧０．０４ＭＰａで通気した。撹拌速度は６００ｒｐｍとした。 To 520 g of salt-free moromi of Example 1, 13 g of potassium dihydrogen phosphate, 0.6 g of antifoaming agent, and 380 mL of tap water were added, adjusted to pH 5.5 with potassium hydroxide, and 115 ° C. for 15 minutes in an autoclave. Was sterilized. Candida utilis was inoculated into this medium and cultured at 30 ° C. for 22 hours according to a conventional method. Separately sterilized 50% glucose aqueous solution was fed at 8.4 g / hr, and aerated air was aerated at 1 L / min and an internal pressure of 0.04 MPa. The stirring speed was 600 rpm.
培養終了後、培養液３０ｍＬにエタノール１ｍＬを添加した後、ｐＨを６．５に調整し、５０℃で２時間振とうした。続いて、ｐＨを５に調整し、ヌクレアーゼアマノＧ（アマノエンザイム社製）を１５ｍｇ加え、７０℃で１時間反応させた。反応後、ｐＨを６．５に調整し、デアミザイム（アマノエンザイム社製）を１５ｍｇ加え、５５℃で１時間反応させた。最後に８５℃で２０分間加温して酵素を失活させ、遠心分離によって固形分を取り除き、核酸発酵調味液（サンプル１７）を得た。 After completion of the culture, 1 mL of ethanol was added to 30 mL of the culture solution, the pH was adjusted to 6.5, and the mixture was shaken at 50 ° C. for 2 hours. Subsequently, the pH was adjusted to 5, 15 mg of nuclease Amano G (manufactured by Amano Enzyme) was added, and the mixture was reacted at 70 ° C. for 1 hour. After the reaction, the pH was adjusted to 6.5, 15 mg of deamizyme (manufactured by Amano Enzyme) was added, and the mixture was reacted at 55 ° C. for 1 hour. Finally, the enzyme was inactivated by heating at 85 ° C. for 20 minutes, the solid content was removed by centrifugation, and a nucleic acid fermentation seasoning liquid (sample 17) was obtained.
アロマイルド（商標、興人ライフサイエンス社製）０．２５ｇを５０ｍＬの浄水に添加し、これを０．１％ギ酸含有水で５０倍希釈したものを呈味性核酸の分析に用いた。ＨＤＭＦの定量にはアロマイルド５０ｍｇを５０ｍＬの浄水に添加したものを用いた。ＨＤＭＦの定量と呈味性核酸の定量条件は実施例１に記載した条件の通りである。 Aromad (trademark, manufactured by Kojin Life Science Co., Ltd.) (0.25 g) was added to 50 mL of purified water, which was diluted 50-fold with 0.1% formic acid-containing water and used for analysis of taste nucleic acids. For quantification of HDMF, 50 mg of aromoid added to 50 mL of purified water was used. The conditions for quantifying HDMF and taste nucleic acids are as described in Example 1.
結果を表５に示す。表５に記載されているとおり、市販こいくち醤油は呈味性核酸が検出限界以下であったが、本発明の発酵調味液（サンプル１７）は呈味性核酸を著量含有していた。また、本発明の発酵調味液（サンプル１７）は市販こいくち醤油と同等のＨＤＭＦを含有していたが、市販高核酸酵母エキスからはＨＤＭＦが検出されなかった。
従って、本発酵調味液は無添加で核酸を高含有し、醤油のような香ばしい風味が優れている調味液として提供することができることが判明した。
The results are shown in Table 5. As shown in Table 5, the commercial koikuchi soy sauce had a taste nucleic acid below the detection limit, but the fermented seasoning liquid of the present invention (sample 17) contained a significant amount of the taste nucleic acid. Moreover, although the fermented seasoning liquid (sample 17) of the present invention contained HDMF equivalent to commercially available koikuchi soy sauce, HDMF was not detected from the commercially available high nucleic acid yeast extract.
Therefore, it has been found that the present fermented seasoning liquid can be provided as a seasoning liquid that does not contain any additives and contains a high amount of nucleic acid and has a savory flavor such as soy sauce.
Claims (10)
ＨＤＭＦ２ｐｐｍ（ｗ／ｖ）以上と、
を含有する、発酵調味料であって、
諸味の食塩濃度が４％（ｗ／ｖ）以下である、
発酵調味料。 10 ppm (w / v) or more of nucleic acid (excluding nucleic acids derived from addition),
HDMF 2 ppm (w / v) or more,
Containing, a fermented seasoning,
The salt concentration of moromi is 4% (w / v) or less,
Fermented seasoning .
大豆又は小麦等を主原料とした穀物原料に、麹菌を接種して固体麹を調製し、水又は食塩水を加えて食塩濃度４％（ｗ／ｖ）以下の諸味を加温分解する工程と、
前記諸味を加熱処理してホスファターゼを失活させることにより殺菌諸味を調製する工程と、
前記殺菌諸味に核酸生産微生物を接種して危害微生物の混入を抑制できる容器で核酸発酵を行う工程と、
前記核酸生産微生物を自己消化して菌体内のＲＮＡを諸味中に遊離させることによりＲＮＡ抽出物を調製する工程と、
前記ＲＮＡ抽出物にヌクレアーゼを作用させて呈味性のある５’−ヌクレオチド類に変換する工程と、
を有する、発酵調味料の製造方法。 It is a manufacturing method of the fermented seasoning of Claim 1, Comprising:
A step of inoculating a koji mold with a cereal raw material made mainly of soybeans or wheat to prepare solid koji, and adding water or saline to thermally decompose moromi with a salt concentration of 4% (w / v) or less; ,
Preparing a sterilized moromi by heat-treating the moromi to inactivate phosphatase;
Inoculating the sterilizing moromi with nucleic acid-producing microorganisms and performing nucleic acid fermentation in a container capable of suppressing contamination of harmful microorganisms;
A step of preparing an RNA extract by self-digesting the nucleic acid-producing microorganisms to release RNA in the cells into moromi;
Converting the RNA extract into tasty 5′-nucleotides by allowing nuclease to act;
A method for producing a fermented seasoning.
The food / beverage products which improved the umami | taste and richness containing the fermented seasoning of any one of Claims 1-4.
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