JP2010252660A - Seaweed fermented composition and method for producing the same - Google Patents

Seaweed fermented composition and method for producing the same Download PDF

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JP2010252660A
JP2010252660A JP2009104819A JP2009104819A JP2010252660A JP 2010252660 A JP2010252660 A JP 2010252660A JP 2009104819 A JP2009104819 A JP 2009104819A JP 2009104819 A JP2009104819 A JP 2009104819A JP 2010252660 A JP2010252660 A JP 2010252660A
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seaweed
lactic acid
fermentation
composition
protease
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JP5467433B2 (en
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Keisuke Furukawa
圭介 古川
Megumi Kawashima
めぐみ 川島
Kenji Aoyama
顕司 青山
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Kikkoman Corp
キッコーマン株式会社
Riken Vitamin Co Ltd
理研ビタミン株式会社
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Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for producing a seaweed fermented composition richly containing an amino acid and also having favorable flavor. <P>SOLUTION: The method for producing the seaweed fermented composition comprises drying seaweed followed by crushing the dried seaweed into powder, and thereafter treating seaweed raw material solution obtained by dissolving the powdered seaweed in water solution such as water with protease followed by fermenting the thus-treated seaweed raw material solution using lactobacillus. The seaweed fermented composition produced by the method is rich in an amino acid content, has favorable flavor and also high safety because of being derived from a natural substance, and is usable as a seasoning or functional food raw material. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

本発明は、海藻類(ワカメ、コンブ)をプロテアーゼで処理し、乳酸菌で発酵させて得られる海藻発酵組成物およびその製造方法に関する。   The present invention relates to a seaweed fermented composition obtained by treating seaweeds (seaweed, kombu) with protease and fermenting them with lactic acid bacteria, and a method for producing the same.

発酵食品は、広義には、微生物の持つ働きを利用し、食物の成分を好ましい物質に変えた食品であり、微生物の作用により食品の保存性向上、栄養価や消化吸収性の向上、あるいは機能性を有する成分の増加等という効果をもたらす利点を有する。また、発酵を経ることにより、食品の素材そのものからは得られない独特の香りや味が付加された結果、風味豊かな発酵食品を提供できる利点を有する。
発酵食品には、有史以前からの長い歴史があり、その優れた利点から、世界各地に多種多様な発酵食品が発展してきた。発酵食品の例としては、チーズやヨーグルト、ワイン、清酒、味噌、醤油、魚醤、納豆、糠漬け、ザワークラウト、キムチ、鮒寿司、豆腐よう等が挙げられ、今日の食卓において欠かせないものとなっている。
Fermented food, in a broad sense, is a food that uses the action of microorganisms to change the ingredients of food to preferred substances. The action of microorganisms improves food preservation, improves nutritional value and digestibility, or functions. It has the advantage of bringing about an effect such as an increase in components having properties. In addition, as a result of adding a unique scent and taste that cannot be obtained from the food material itself through fermentation, there is an advantage that a fermented food rich in flavor can be provided.
Fermented foods have a long history since prehistoric times, and a wide variety of fermented foods have been developed around the world because of their excellent advantages. Examples of fermented foods include cheese, yogurt, wine, sake, miso, soy sauce, fish sauce, natto, pickled cucumber, sauerkraut, kimchi, sushi, and tofu. ing.

しかし、上記のように、牛乳、果実、米、大豆、魚介類、野菜、豆腐等、幅広い食材を原料とした発酵食品が広く知られているにも関わらず、海藻類を主原料とする発酵食品の開発は未だ十分に進んでいるとはいえない。   However, as mentioned above, although fermented foods made from a wide range of ingredients such as milk, fruit, rice, soybeans, seafood, vegetables, and tofu are widely known, fermentation using seaweed as the main ingredient Food development is still not fully advanced.

海藻は、例えば、佃煮や、汁物、煮物、サラダ等の具材として用いられる他にも、コンブ等が旨味成分の「ダシ」として用いられるように、旨味成分を多く含むことが古くから知られている。さらに海藻はミネラル、食物繊維等を豊富に含有し、近年では、血圧低下作用、免疫調節機能増強作用等を有する成分が見出される等、機能性食品素材の原料としての側面も注目されている。従って、海藻類を原料とする発酵食品の開発により、新たな風味、栄養成分または機能性を有する魅力的な発酵食品を新たに提供できることが大いに期待される。   Seaweed has long been known to contain many umami ingredients, such as koji, soup, boiled food, salads, and other ingredients, such as kombu. ing. Furthermore, seaweed contains abundant minerals, dietary fiber, and the like, and in recent years, it has been attracting attention as a raw material for functional food materials, such as the discovery of components having a blood pressure lowering effect, an immunoregulatory function enhancing effect, and the like. Therefore, it is highly expected that an attractive fermented food having a new flavor, nutritional component or functionality can be newly provided by developing a fermented food using seaweed as a raw material.

従来、海藻を発酵させる技術としては、海藻中に含まれる成分を水などで抽出後、乳酸菌または麹菌で発酵させる方法(例えば、特許文献1参照)や、コンブエキスを含む原料を乳酸菌としてLactobacillus hilgardiiを用いて発酵させた後、一定量のGABAを添加することで、発酵物が持つ特有のエグ味や苦味を抑えた飲食品を提供する方法が知られている(例えば、特許文献2参照)。しかしながら、海藻を単に乳酸菌と接触させても、海藻中に含まれるセルロース等の多糖類により発酵が阻害され、充分に発酵が進行せず、この方法では、海藻中に含まれる各種成分を抽出する工程を経なければならないため、海藻中に含まれる豊富な糖やタンパク質を有効に利用できないといった問題が生じてしまう。その欠点を改善するために、海藻を発酵する前にセルラーゼで処理することにより、発酵を阻害する多糖類を分解する方法(例えば、特許文献3〜5参照)が提案されている。この方法によれば、セルラーゼの作用により、特に糖質を効率的に遊離することができ、栄養価が豊富な発酵食品を得ることができ、あるいは栄養価の高い飼料を提供することができる。一方、発酵食品の旨味や風味に関与するグルタミン酸等のアミノ酸を海藻原料から十分に引き出すという観点においては、未だ技術開発の余地がみられる。特に、褐藻類では発酵が進行しにくく、従来の方法によっては、海藻中に含まれるアミノ酸を十分に遊離させることが困難であるのが現状である。   Conventionally, as a technique for fermenting seaweed, a method in which components contained in seaweed are extracted with water or the like and then fermented with lactic acid bacteria or koji molds (see, for example, Patent Document 1), or a material containing kumbu extract as a lactic acid bacterium, Lactobacillus hilgardii A method of providing a food or drink with reduced fermented product's peculiar taste and bitterness is known by adding a certain amount of GABA after fermenting with sucrose (see, for example, Patent Document 2). . However, even when seaweed is simply brought into contact with lactic acid bacteria, fermentation is inhibited by polysaccharides such as cellulose contained in seaweed and fermentation does not proceed sufficiently. In this method, various components contained in seaweed are extracted. Since the process must be performed, there arises a problem that abundant sugars and proteins contained in seaweed cannot be used effectively. In order to improve the defect, a method of degrading polysaccharides that inhibit fermentation by treating with cellulase before fermenting seaweeds (see, for example, Patent Documents 3 to 5) has been proposed. According to this method, due to the action of cellulase, particularly carbohydrates can be efficiently released, fermented foods rich in nutritional value can be obtained, or feed with high nutritional value can be provided. On the other hand, there is still room for technological development in terms of sufficiently extracting amino acids such as glutamic acid involved in the umami and flavor of fermented foods from seaweed materials. In particular, in brown algae, fermentation is unlikely to proceed, and depending on conventional methods, it is difficult to sufficiently release amino acids contained in seaweed.

すなわち、海藻類を発酵させた風味の良い海藻発酵組成物および海藻中の成分、特にアミノ酸類をより多く遊離でき、同じ量の原料からより多くの旨味成分や機能性成分が得られ、より短時間の工程で旨味成分や機能性成分を効率よく遊離・変換することが可能な海藻発酵組成物の製造方法が求められている。   In other words, a flavorful seaweed fermentation composition obtained by fermenting seaweeds and components in seaweed, particularly amino acids, can be released more, and more umami components and functional components can be obtained from the same amount of raw materials. There is a need for a method for producing a seaweed fermentation composition that can efficiently release and convert umami components and functional components in a time step.

特開平3−172147号公報Japanese Patent Laid-Open No. 3-172147 特許第4128893号明細書Japanese Patent No. 4128893 特開平1−277482号公報JP-A-1-277482 特許第3637353号明細書Japanese Patent No. 3637353 特許第4017783号明細書Japanese Patent No. 4017783

本発明が解決しようとする課題は、アミノ酸量に富み、かつ、良好な風味を有する海藻発酵組成物を提供することであり、また、その効率的な製造方法を提供することにある。   The problem to be solved by the present invention is to provide a seaweed fermentation composition that is rich in amino acids and has a good flavor, and to provide an efficient production method thereof.

本発明者らは上記課題を解決すべく、鋭意研究を重ねた結果、海藻類の粉末を含む水溶液をプロテアーゼで処理することにより、セルラーゼ処理を行うよりもグルコース遊離率およびタンパク遊離率が高くなっていることが分かった。そして、さらに、鋭意検討した結果、海藻類の粉末1〜3%を含む水溶液に、終濃度0.5〜1.0%となるようプロテアーゼを加えて30℃で48時間培養した後、または、酵素処理と同時に乳酸菌を添加し、30℃で48時間発酵することにより、従来の方法に比べ、アミノ酸生産量およびL−乳酸生産量が増加し、かつ、良好な風味を呈する海藻発酵組成物が得られることを知り、本発明を完成させた。   As a result of intensive studies to solve the above problems, the inventors of the present invention have increased the glucose release rate and the protein release rate by treating an aqueous solution containing seaweed powder with protease, compared to cellulase treatment. I found out. And, as a result of further intensive studies, after adding protease to an aqueous solution containing 1 to 3% of seaweed powder to a final concentration of 0.5 to 1.0% and culturing at 30 ° C. for 48 hours, or By adding lactic acid bacteria simultaneously with the enzyme treatment and fermenting at 30 ° C. for 48 hours, a seaweed fermentation composition having an increased amino acid production amount and L-lactic acid production amount and a good flavor as compared with the conventional method is obtained. Knowing that it can be obtained, the present invention has been completed.

すなわち、本発明は、
1) 海藻類をプロテアーゼで処理し、乳酸菌により発酵させることを特徴とする海藻発酵組成物。
2) 乳酸菌が、Pediococcus acidilactici、Lactbacillus curvatus、Leuconostoc mesenteroides subsp.mesenteroidesまたはPediococcus pentosaceusから選択される1以上である上記1記載の海藻発酵組成物。
3) グルタミン酸を20mg/dl以上、乳酸を70mg/dl以上含有する、上記1または上記2記載の海藻発酵組成物。
4) 海藻類をプロテアーゼで処理し、乳酸菌により発酵させることを特徴とする海藻発酵組成物の製造方法。
5) 上記1、2または3記載の海藻発酵組成物を利用して得られる飲食品。
を提供するものである。
That is, the present invention
1) A seaweed fermentation composition characterized in that seaweed is treated with a protease and fermented with lactic acid bacteria.
2) The seaweed fermentation composition according to 1 above, wherein the lactic acid bacterium is one or more selected from Pediococcus acidilactici, Lactbacillus curvatus, Leuconostoc mesenteroides subsp. Mesenteroides, or Pediococcus pentosaceus.
3) The seaweed fermentation composition according to 1 or 2 above, containing glutamic acid at 20 mg / dl or more and lactic acid at 70 mg / dl or more.
4) A method for producing a seaweed fermentation composition comprising treating seaweed with a protease and fermenting with a lactic acid bacterium.
5) A food or drink obtained using the seaweed fermentation composition according to 1, 2 or 3.
Is to provide.

本発明により、アミノ酸量に富み、かつ、良好な風味を有する海藻発酵組成物を提供することができる。本発明の海藻発酵組成物は、天然物由来であるため安全性が高く、調味料、機能性食品素材として利用することが可能である。   ADVANTAGE OF THE INVENTION By this invention, the seaweed fermentation composition which is rich in the amount of amino acids, and has favorable flavor can be provided. Since the seaweed fermentation composition of the present invention is derived from a natural product, it is highly safe and can be used as a seasoning or a functional food material.

以下、本発明を具体的に説明する。
(海藻類)
The present invention will be specifically described below.
(Seaweed)

本発明における海藻類は、特に限定されることはないが、褐藻類、緑藻類および紅藻類からなる群から選ばれる少なくとも一種であることが好ましい。具体的には、アオサ、アオノリ、テングサ、コンブ、ワカメ、アラメ、ホンダワラ、アカモク、ヒジキ等があげられ、中でも、コンブ、ワカメは、海藻中に含まれる炭素源および窒素源が豊富で、かつ、安価で、容易に入手できるという点で好ましい。これらの海藻は、生のまま、乾燥品として、または乾燥粉末として用いることができる。
(海藻原料液)
The seaweed in the present invention is not particularly limited, but is preferably at least one selected from the group consisting of brown algae, green algae and red algae. Specifically, Aosa, Aonori, Proboscis, Kombu, Wakame, Alame, Honda Walla, Akamoku, Hijiki, etc., among them, Kombu and Wakame are rich in carbon and nitrogen sources contained in seaweed, and It is preferable in that it is inexpensive and easily available. These seaweeds can be used raw, as a dry product, or as a dry powder.
(Seaweed raw material liquid)

海藻類は、酵素処理および乳酸菌での分解を容易に進行させるために、乾燥後に破砕し粉末状にした後、水等の液体に溶解させ、水溶液(以下、「海藻原料液」という。)として用いることが好ましい。海藻類を用いる濃度は特に規定しないが、風味に富んだ海藻発酵組成物を得るために、質量パーセント濃度換算で1〜4%の粉末状の海藻類を原料として用いることが好ましい。4%を越える粉末状の海藻類を原料として用いる場合、粉末が膨潤し、固形状となってしまい、発酵が充分に進行しない。一方で、1%未満の粉末状の海藻類を原料として用いる場合、海藻自体がもつタンパク、アミノ酸量が少ないため、発酵物の味が希薄になるといった欠点がある。
(酵素分解)
In order to facilitate the enzymatic treatment and degradation with lactic acid bacteria, seaweeds are crushed and powdered after drying, and then dissolved in a liquid such as water to obtain an aqueous solution (hereinafter referred to as “seaweed raw material liquid”). It is preferable to use it. The concentration at which seaweed is used is not particularly defined, but in order to obtain a seaweed fermentation composition rich in flavor, it is preferable to use 1 to 4% of powdered seaweed as a raw material in terms of mass percent concentration. When powdery seaweeds exceeding 4% are used as raw materials, the powder swells and becomes solid, and the fermentation does not proceed sufficiently. On the other hand, when using less than 1% of powdery seaweed as a raw material, there is a disadvantage that the taste of the fermented product becomes dilute because the amount of protein and amino acid of seaweed itself is small.
(Enzymatic degradation)

酵素分解は、海藻類を含む水溶液にプロテアーゼを添加、加温することで行う。使用するプロテアーゼは安価に入手できる食品添加物用のプロテアーゼを用いることができる。特に、かび由来のプロテアーゼは力価が高く、少量の添加で効果があるため好ましい。海藻原料液を用いて発酵を行う場合のプロテアーゼの濃度は、例えば食品添加用のプロテアーゼ(例えば、天野エンザイム社製)を用いる場合、0.5〜1.0%の濃度の範囲が好ましい。1.0%以上の濃度のプロテアーゼを用いても、添加量の増加に伴う効果に大きな変化がなく、意味がない。0.5%以下の濃度のプロテアーゼを用いる場合、グルコース遊離量およびタンパク遊離量が低く、良好な発酵組成物が得られなくなってしまう。
(乳酸発酵)
Enzymatic degradation is performed by adding protease to an aqueous solution containing seaweed and heating. As the protease to be used, a protease for food additives that can be obtained at low cost can be used. In particular, fungi-derived protease is preferable because it has a high titer and is effective when added in a small amount. The concentration of the protease in the case of performing fermentation using the seaweed raw material liquid is preferably in the range of 0.5 to 1.0% when, for example, a protease for food addition (for example, Amano Enzyme) is used. Even if a protease having a concentration of 1.0% or more is used, there is no significant change in the effect accompanying an increase in the amount of addition, which is meaningless. When a protease having a concentration of 0.5% or less is used, the amount of released glucose and the amount of released protein are low, and a good fermentation composition cannot be obtained.
(Lactic acid fermentation)

本発明に用いる乳酸菌は、ラクトバチルス属(Lactobacillus)、ラクトコッカス属(Lactococcus)、ストレプトコッカス属(Streptcoccus)、ビフィドバクテリウム属(Bifidobacterium)、ロイコノストック属(Leuconostoc)などを用いることができ、これらのうち一または二以上の乳酸菌を選択し、同時または逐次的に使用できる。中でも、発酵によるアミノ酸生成という点で、ラクトバチルス属、ロイコノストック属を用いることが好ましい。特に、Pediococcus acidilactici(NRIC 0124)、Lactobacillus curvatus(NRIC 1975)、Leuconostoc mesenteroides subsp. mesenteroides(NRIC 1087)およびPediococcus pentosaceus(NRIC 0122)は、様々な海藻類を効率よく発酵することができ、最も好ましい。乳酸発酵は、酵素分解を行った後に行うことで、好ましい栄養価や風味を持った発酵組成物が得られるが、酵素分解と同時に行うこともできる。尚、乳酸発酵を酵素分解と同時に行う場合、添加する酵素由来の雑菌によって汚染することが考えられるため、酵素溶液をあらかじめフィルター除菌しておくか、または乳酸菌の接種量を増やし、水溶液中のpHを速やかに低下させることで、雑菌の増殖を防ぐ等の操作が必要である。
(発酵組成物)
Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Streptococcus (Streptcoccus), Bifidobacterium (Leuconostoc), and the like can be used as the lactic acid bacteria used in the present invention, Of these, one or more lactic acid bacteria can be selected and used simultaneously or sequentially. Especially, it is preferable to use the Lactobacillus genus and Leuconostoc genus at the point of the amino acid production | generation by fermentation. In particular, Pediococcus acidilactici (NRIC 0124), Lactobacillus curvatus (NRIC 1975), Leuconostoc mesenteroides subsp. Mesenteroides (NRIC 1087) and Pediococcus pentosaceus (NRIC 0122) are most preferable because they can efficiently ferment various seaweeds. Lactic acid fermentation is performed after enzymatic degradation, whereby a fermentation composition having a favorable nutritional value and flavor can be obtained. However, it can also be performed simultaneously with enzymatic degradation. In addition, when performing lactic acid fermentation at the same time as enzymatic degradation, it may be contaminated by the bacteria derived from the enzyme to be added. Therefore, pre-filter the enzyme solution or increase the amount of lactic acid bacteria inoculated, By rapidly lowering the pH, an operation such as preventing the growth of various bacteria is required.
(Fermented composition)

海藻類をプロテアーゼ処理後、乳酸菌を用いて発酵し、得られる組成物(以下、「海藻発酵組成物」という。)は、海藻由来のアミノ酸を利用し、そのまま調味料等に使用することもできるが、固形分をろ過や遠心分離によって除去し、上清を抽出した後、醤油のように使用したり、ドレッシングや麺つゆ等の成分のひとつとしても使用することができる。また、海藻発酵組成物は、GABA等の機能性組成物を豊富に含有しているため、そのまま健康飲料の成分とするか、濃縮、乾燥したうえで健康食品の成分として利用することが可能である。   The seaweed is treated with protease and then fermented with lactic acid bacteria, and the resulting composition (hereinafter referred to as “seaweed fermentation composition”) can be used as a seasoning as it is, using amino acids derived from seaweed. However, after removing the solid content by filtration or centrifugation and extracting the supernatant, it can be used like soy sauce or used as one of ingredients such as dressing or noodle soup. In addition, since the seaweed fermentation composition contains abundant functional compositions such as GABA, it can be used as a component of a health drink as it is or as a component of a health food after being concentrated and dried. is there.

以下に本発明の実施例を記載するが、本発明はこれらに何ら限定されるものではない。   Examples of the present invention will be described below, but the present invention is not limited thereto.

[各種乳酸菌を用いた海藻発酵組成物の調製]
食品に広く用いられている乳酸菌487株を用いて、海藻発酵組成物を調整した。各乳酸菌を、40mlのMRS培地にて30℃で20時間静置培養後、遠心分離にて菌体を分取し、それぞれに30%グリセリンを添加した50%MRS培地を加えて、菌体懸濁液を得た。次に、粉末ワカメ(理研ビタミン社製)200mgおよびプロテアーゼ(天野エンザイム社製)200mgに、滅菌水を総量20mlとなるよう加えて得られる粉末ワカメ水溶液20mlに、菌体懸濁液0.1mlを加え、30℃で48時間発酵し、発酵組成物を得た。各海藻発酵組成物のL−乳酸量およびグルタミン酸量を、バイオセンサーBF−5(王子計測社製)を用いて測定した。その代表例を表1に示す。
[Preparation of seaweed fermentation composition using various lactic acid bacteria]
A seaweed fermentation composition was prepared using lactic acid bacteria 487 strain widely used in foods. Each lactic acid bacterium is cultivated by standing still in 40 ml of MRS medium at 30 ° C. for 20 hours, and then the microbial cells are collected by centrifugation, and 50% MRS medium added with 30% glycerin is added to each lactic acid bacterium. A turbid liquid was obtained. Next, 0.1 ml of a cell suspension is added to 20 ml of a powdered wakame aqueous solution obtained by adding sterile water to 200 mg of powdered wakame (manufactured by Riken Vitamin) and 200 mg of protease (manufactured by Amano Enzyme) so that the total amount becomes 20 ml. In addition, fermentation was performed at 30 ° C. for 48 hours to obtain a fermented composition. The amount of L-lactic acid and the amount of glutamic acid in each seaweed fermentation composition were measured using a biosensor BF-5 (manufactured by Oji Scientific Instruments). A typical example is shown in Table 1.

表1で示すとおり、使用する菌株によって、得られる海藻発酵組成物の組成には差がみられた。   As shown in Table 1, there was a difference in the composition of the obtained seaweed fermentation composition depending on the strain used.

次いで、海藻発酵組成物中に含まれるアミノ酸量を測定した。本発明である海藻発酵組成物は、前記の方法に準じて、プロテアーゼを加えた粉末ワカメ水溶液に、乳酸菌としてPediococcus acidilactici(本発明1)、Lactobacillus curvatus(本発明2)およびLeuconostoc mesenteroides subsp. mesenteroides(本発明3)をそれぞれ添加し、30℃で48時間発酵させることで得た。アミノ酸生成能の測定は、当該発酵組成物の遠心上清に対し、HPLCによるアミノ酸分析を行うことで測定した。比較例として、プロテアーゼを加えた粉末ワカメ水溶液に乳酸菌を添加せずに、30℃で48時間加温した未発酵の組成物(比較例1)を用い、前記の方法に従ってアミノ酸分析を行った。各乳酸菌を用いて得た海藻発酵組成物および比較例中の遊離アミノ酸の値を表2に示す。   Subsequently, the amount of amino acids contained in the seaweed fermentation composition was measured. According to the method described above, the seaweed fermentation composition according to the present invention is added to a powdered wakame aqueous solution to which a protease has been added. The present invention 3) was added and fermented at 30 ° C. for 48 hours. The amino acid production ability was measured by performing amino acid analysis by HPLC on the centrifugal supernatant of the fermentation composition. As a comparative example, amino acid analysis was performed according to the above method using an unfermented composition (Comparative Example 1) heated at 30 ° C. for 48 hours without adding lactic acid bacteria to a powdered wakame aqueous solution to which protease was added. Table 2 shows the values of free amino acids in the seaweed fermentation composition and comparative examples obtained using each lactic acid bacterium.

表2に示すとおり、本発明である海藻発酵組成物は、比較例である乳酸菌による発酵を行わなかった組成物に比べ、総遊離アミノ酸量が高かった。また、各種遊離アミノ酸の量も、本発明である海藻発酵組成物の含有量が、比較例に対して一様に高くなり、特に、うまみに関連するグルタミン酸、アラニン、セリン、スレオニン、グリシン、アスパラギン酸等のアミノ酸が顕著に増加した。ここで見出された、アミノ酸遊離量の多い菌株Leuconostoc mesenteroides subsp.mesenteroidesを、実施例2、3および4で使用した。   As shown in Table 2, the seaweed fermentation composition of the present invention had a higher total free amino acid amount than the composition that was not subjected to fermentation with lactic acid bacteria as a comparative example. In addition, the amount of various free amino acids is also uniformly higher than that of the comparative example in the content of the seaweed fermentation composition according to the present invention. Amino acids such as acids increased significantly. The strain Leuconostoc mesenteroides subsp. Mesenteroides found here, which has a high amino acid release, was used in Examples 2, 3 and 4.

[ワカメを用いた海藻発酵組成物の調節の検討]
海藻をプロテアーゼまたはセルラーゼで処理して得られる組成物中に含まれるグルコース、タンパクの遊離量を測定した。
[Examination of adjustment of seaweed fermentation composition using seaweed]
The amount of glucose and protein contained in the composition obtained by treating seaweed with protease or cellulase was measured.

海藻原料液として、粉末ワカメ(理研ビタミン社製)0.2gまたは0.4gを水20mlに加えて撹拌して得られる1%および2%粉末ワカメ水溶液を用いた。各海藻原料液に対して、プロテアーゼ(天野エンザイム社製)またはセルラーゼ(天野エンザイム社製)を終濃度が0.5%、1.0%となるように添加し、30℃で48時間処理した。次に、各酵素処理後の水溶液中に含まれるグルコース量をグルコーステストワコーII(和光純薬社製)を用いて、タンパク量をDCプロテインアッセイ(BIORAD社製)を用いて測定した。各組成物中に含まれるグルコースおよびタンパク遊離量を表3に示す。   As seaweed raw material liquid, 1% and 2% powdered seaweed aqueous solution obtained by adding 0.2 g or 0.4 g of powdered seaweed (manufactured by Riken Vitamin Co., Ltd.) to 20 ml of water and stirring was used. Protease (manufactured by Amano Enzyme) or cellulase (manufactured by Amano Enzyme) was added to each seaweed raw material solution so that the final concentrations were 0.5% and 1.0%, followed by treatment at 30 ° C. for 48 hours. . Next, the amount of glucose contained in the aqueous solution after each enzyme treatment was measured using Glucose Test Wako II (manufactured by Wako Pure Chemical Industries), and the protein amount was measured using DC protein assay (manufactured by BIORAD). Table 3 shows the amount of glucose and protein released in each composition.

表3に示すとおり、粉末ワカメ水溶液をプロテアーゼで処理した場合、セルラーゼ処理を行うよりもグルコースおよびタンパク遊離量が高くなることが確認された。   As shown in Table 3, it was confirmed that when the powdered wakame aqueous solution was treated with protease, glucose and protein release amounts were higher than when cellulase treatment was performed.

グルコースおよびタンパクは、乳酸菌にとって重要な炭素源および窒素源となることから、プロテアーゼ処理物を発酵させた場合、菌体の増殖が良好で、アミノ酸(特にグルタミン酸)に富む発酵物の調整が期待できる。   Glucose and protein are important carbon and nitrogen sources for lactic acid bacteria, so when fermented protease products, the growth of the cells is good and adjustment of fermented products rich in amino acids (particularly glutamic acid) can be expected. .

次に、プロテアーゼ処理後の水溶液に、乳酸菌を添加、培養することで得た海藻発酵組成物中のL−乳酸およびグルタミン酸量を測定した。実施例1に記載の方法に準じ、海藻原料液として全量20mlとなる1%粉末ワカメ水溶液を用い、当該海藻原料液に、終濃度が0.5%または1.0%となるようプロテアーゼを添加し、さらに菌体懸濁液0.1mlを用いて乳酸菌を接種した後、30℃で48時間培養し、本発明である海藻発酵組成物(本発明2、3)を得た。乳酸菌として、Leuconostoc mesenteroides subsp.mesenteroidesを用いた。比較例として、海藻原料液として全量20mlとなる1%および2%粉末ワカメ水溶液を用い、酵素を添加せずに、乳酸菌を接種した後、30℃、48時間の発酵後に得られる組成物(比較例2)および終濃度が0.5%または1.0%となるようセルラーゼを添加し、乳酸菌を接種後、発酵して得られる組成物(比較例3,4)を用いた。発酵後のL−乳酸、グルタミン酸量は、バイオセンサーBF−5(王子計測社製)にて測定した。本発明および比較例中に含まれるL−乳酸およびグルタミン酸量を表4に示す。   Next, the amounts of L-lactic acid and glutamic acid in the seaweed fermentation composition obtained by adding and culturing lactic acid bacteria to the aqueous solution after protease treatment were measured. According to the method described in Example 1, a 1% powdered wakame aqueous solution with a total volume of 20 ml is used as the seaweed raw material solution, and protease is added to the seaweed raw material solution so that the final concentration is 0.5% or 1.0%. Then, after inoculating lactic acid bacteria using 0.1 ml of the cell suspension, the cells were cultured at 30 ° C. for 48 hours to obtain the seaweed fermentation composition (Inventions 2 and 3) according to the present invention. Leuconostoc mesenteroides subsp. Mesenteroides was used as a lactic acid bacterium. As a comparative example, a composition obtained after fermentation at 30 ° C. for 48 hours after inoculating lactic acid bacteria without adding enzymes using 1% and 2% powdered wakame aqueous solution with a total amount of 20 ml as seaweed raw material liquid Example 2) and a composition (Comparative Examples 3 and 4) obtained by adding cellulase to a final concentration of 0.5% or 1.0%, inoculating lactic acid bacteria, and then fermenting were used. The amounts of L-lactic acid and glutamic acid after fermentation were measured with Biosensor BF-5 (manufactured by Oji Scientific Instruments). Table 4 shows the amounts of L-lactic acid and glutamic acid contained in the present invention and comparative examples.

表4に示すとおり、本発明である海藻発酵組成物中に含まれるL−乳酸およびグルタミン酸量は、比較例よりも高く、プロテアーゼを加えることにより海藻の乳酸発酵が良好に進行することが確認された。   As shown in Table 4, the amount of L-lactic acid and glutamic acid contained in the seaweed fermentation composition according to the present invention is higher than that of the comparative example, and it was confirmed that lactic acid fermentation of seaweed proceeds well by adding protease. It was.

また、海藻原料液として、2%粉末ワカメ水溶液を用いた場合の結果を表5に示す。本発明である海藻発酵組成物(本発明4,5)および比較例である各組成物(比較例5〜7)は、前記と同様の方法で得た。   Table 5 shows the results when a 2% powdered seaweed aqueous solution was used as the seaweed raw material liquid. The seaweed fermentation composition according to the present invention (present inventions 4 and 5) and the compositions according to comparative examples (comparative examples 5 to 7) were obtained by the same method as described above.

表5に示すとおり、本発明である海藻発酵組成物は、海藻原料液中の原料濃度によらず、海藻の発酵が良好に進行することが確認された。   As shown in Table 5, it was confirmed that the seaweed fermentation composition according to the present invention favorably proceeds with seaweed fermentation regardless of the raw material concentration in the seaweed raw material liquid.

[コンブを用いた海藻発酵組成物の調節の検討]
海藻原料液として、粉砕した干しコンブ(南茅部産)0.2gまたは0.6gを水20mlに加えて撹拌して1%および3%粉末コンブ水溶液を作製し、実施例2に記載の方法に従い、各水溶液をプロテアーゼまたはセルラーゼで処理後、各酵素処理後の水溶液中に含まれるグルコースおよびタンパクの遊離量を算出した。その結果を表6に示す。
[Examination of adjustment of seaweed fermentation composition using kombu]
As seaweed raw material liquid, 0.2 g or 0.6 g of pulverized dried kombu (produced from Nanbu) was added to 20 ml of water and stirred to prepare 1% and 3% powdered kombu aqueous solution. According to the method described in Example 2, After each aqueous solution was treated with protease or cellulase, the amount of glucose and protein released in the aqueous solution after each enzyme treatment was calculated. The results are shown in Table 6.

表6に示すとおり、粉末コンブ水溶液をプロテアーゼで処理した場合、セルラーゼ処理と比較してグルコース量は同等でタンパク遊離量が2倍程度高くなることが確認された。   As shown in Table 6, it was confirmed that when the powdered comb aqueous solution was treated with protease, the amount of glucose was the same and the protein release amount was about twice as high as that of the cellulase treatment.

次いで、各種酵素で処理した水溶液で乳酸発酵を行い、L−乳酸およびグルタミン酸量を測定した。その結果を表7に示す。   Next, lactic acid fermentation was performed with aqueous solutions treated with various enzymes, and the amounts of L-lactic acid and glutamic acid were measured. The results are shown in Table 7.

表7に示すとおり、本発明である海藻発酵組成物中に含まれるL−乳酸およびグルタミン酸量は、比較例よりも高く、プロテアーゼを加えることにより海藻の乳酸発酵が良好に進行することが確認された。この結果から、原料としてワカメを用いた場合だけでなく、コンブを用いた場合でもプロテアーゼ処理後に乳酸発酵を行うことで、良好な発酵状態となり、発酵後の海藻発酵組成物中に含まれるL−乳酸およびグルタミン酸量が従来のものに比べて高くなることが確認された。この方法により、各種海藻類において広く汎用が可能な海藻発酵組成物を得ることが期待できる。   As shown in Table 7, the amount of L-lactic acid and glutamic acid contained in the seaweed fermentation composition according to the present invention is higher than that of the comparative example, and it was confirmed that lactic acid fermentation of seaweed proceeds well by adding protease. It was. From this result, not only when using seaweed as a raw material, but also when using kombu, lactic acid fermentation is performed after protease treatment, so that a good fermentation state is achieved, and L- contained in the seaweed fermentation composition after fermentation It was confirmed that the amount of lactic acid and glutamic acid was higher than that of the conventional one. By this method, it can be expected to obtain a seaweed fermentation composition that can be widely used in various seaweeds.

[セルラーゼまたはプロテアーゼを用いた海藻発酵組成物含有アミノ酸量の比較]
粉末ワカメ(理研ビタミン社製)0.2gまたは粉砕した干しコンブ(南茅部産)0.2gを、水20mlにそれぞれ加えて撹拌した。当該海藻原料液に、終濃度が1.0%となるようプロテアーゼを添加した後、さらに菌体懸濁液0.1mlを用いて乳酸菌を接種した後、30℃で48時間培養し、本発明である海藻発酵組成物(本発明8、9)を得た。乳酸菌としては、Leuconostoc mesenteroides subsp.mesenteroidesを用いた。比較例としては、海藻原料液として全量20mlとなる1%粉末ワカメ水溶液、1%粉末コンブ水溶液を用い、セルラーゼまたはプロテアーゼを終濃度1%となるよう添加し、乳酸菌を接種せずに、30℃、48時間の加温後に得られる組成物(比較例11,13,14,16)および各海藻原料液に終濃度が1.0%なるようセルラーゼを添加し、乳酸菌を接種後、30℃で48時間発酵して得られる組成物(比較例12,15)を用いた。アミノ酸量は、実施例1に記載の方法と同様に測定した。
[Comparison of amino acid content in seaweed fermentation composition using cellulase or protease]
0.2 g of powdered seaweed (manufactured by Riken Vitamin Co., Ltd.) or 0.2 g of crushed dried kombu (from Nantobe) was added to 20 ml of water and stirred. Protease is added to the seaweed raw material solution so that the final concentration is 1.0%, and further lactic acid bacteria are inoculated using 0.1 ml of the cell suspension, followed by culturing at 30 ° C. for 48 hours. A seaweed fermentation composition (Inventions 8 and 9) was obtained. Leuconostoc mesenteroides subsp. Mesenteroides was used as the lactic acid bacterium. As a comparative example, using a 1% powdered seaweed aqueous solution and a 1% powdered kombu aqueous solution in a total volume of 20 ml as a seaweed raw material solution, cellulase or protease was added to a final concentration of 1%, and 30 ° C. without inoculating lactic acid bacteria. Cellulase was added to the composition obtained after 48 hours of heating (Comparative Examples 11, 13, 14, 16) and each seaweed raw material solution to a final concentration of 1.0%, and after inoculation with lactic acid bacteria, at 30 ° C. A composition obtained by fermentation for 48 hours (Comparative Examples 12 and 15) was used. The amount of amino acid was measured in the same manner as described in Example 1.

表8に示すとおり、本発明である海藻発酵組成物(本発明8,9)は、各アミノ酸を豊富に含有することが確認された。また、プロテアーゼ処理のみで、発酵を行わなかった組成物(比較例13,16)に対し、遊離アミノ酸量は約1.5倍に、セルラーゼ処理後に発酵して得られる組成物(比較例12,15)に対し、遊離アミノ酸量は約6倍に増加していた。本発明である海藻発酵組成物は、乳酸菌により海藻原料液が効率よく発酵され、旨味成分であるグルタミン酸等のアミノ酸を豊富に含有していることから、従来の海藻を発酵して得られる組成物に比べ、風味に優れていることが期待される。   As shown in Table 8, it was confirmed that the seaweed fermentation composition according to the present invention (present inventions 8 and 9) contains abundant amino acids. In addition, a composition obtained by fermentation after cellulase treatment (Comparative Example 12, 16), the amount of free amino acid is about 1.5 times that of the composition (Comparative Examples 13 and 16) that was treated only with protease and not fermented. In contrast to 15), the amount of free amino acid increased about 6 times. The seaweed fermentation composition according to the present invention is a composition obtained by fermenting conventional seaweed because the seaweed raw material liquid is efficiently fermented by lactic acid bacteria and contains abundant amino acids such as glutamic acid, which is an umami component. It is expected to have a better flavor than

[官能評価試験]
海藻発酵組成物の官能評価試験を行った。官能評価試験に用いた本発明の海藻発酵組成物および比較例の組成物は、実施例2に記載の方法で得た。官能評価試験は、5人のパネラーに対するブラインドテストにより実施し、酸味および旨味についての評価を行った。評価は、各サンプルの相対的な評価を平均化することで算出した。その結果を表9に示す。
[Sensory evaluation test]
The sensory evaluation test of the seaweed fermentation composition was performed. The seaweed fermentation composition of the present invention used in the sensory evaluation test and the composition of the comparative example were obtained by the method described in Example 2. The sensory evaluation test was carried out by a blind test for five panelists, and the sourness and umami were evaluated. Evaluation was calculated by averaging the relative evaluation of each sample. The results are shown in Table 9.

表9に示すとおり、本発明である海藻発酵組成物は、セルラーゼ処理後に乳酸菌により発酵してえられる組成物に比べて、酸味および旨味に優れていることが確認された。   As shown in Table 9, it was confirmed that the seaweed fermentation composition according to the present invention is superior in sourness and umami compared to a composition obtained by fermentation with lactic acid bacteria after cellulase treatment.

Claims (5)

海藻類をプロテアーゼで処理し、乳酸菌により発酵させることを特徴とする海藻発酵組成物。 A seaweed fermentation composition, wherein seaweed is treated with a protease and fermented with lactic acid bacteria. 乳酸菌が、Pediococcus acidilactici、Lactbacillus curvatus、Leuconostoc mesenteroides subsp.mesenteroidesまたはPediococcus pentosaceusから選択される1以上である請求項1記載の海藻発酵組成物。 The seaweed fermentation composition according to claim 1, wherein the lactic acid bacterium is one or more selected from Pediococcus acidilactici, Lactbacillus curvatus, Leuconostoc mesenteroides subsp. Mesenteroides, or Pediococcus pentosaceus. グルタミン酸を20mg/dl以上、乳酸を70mg/dl以上含有する、請求項1または請求項2記載の海藻発酵組成物。 The seaweed fermentation composition according to claim 1 or 2, comprising glutamic acid at 20 mg / dl or more and lactic acid at 70 mg / dl or more. 海藻類をプロテアーゼで処理し、乳酸菌により発酵させることを特徴とする海藻発酵組成物の製造方法。 A method for producing a seaweed fermentation composition comprising treating seaweed with a protease and fermenting with a lactic acid bacterium. 請求項1、2または3に記載の海藻発酵組成物を利用して得られる飲食品。 The food-drinks obtained using the seaweed fermentation composition of Claim 1, 2 or 3.
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