JP4115453B2 - Yeast extract and production method thereof - Google Patents
Yeast extract and production method thereof Download PDFInfo
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- JP4115453B2 JP4115453B2 JP2005024258A JP2005024258A JP4115453B2 JP 4115453 B2 JP4115453 B2 JP 4115453B2 JP 2005024258 A JP2005024258 A JP 2005024258A JP 2005024258 A JP2005024258 A JP 2005024258A JP 4115453 B2 JP4115453 B2 JP 4115453B2
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- yeast extract
- peptide
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Images
Description
本発明は、酵母エキスおよびその製造方法に関し、詳しくはコク味付与能、マスキング能に優れた酵母エキスおよびその製造法に関するものである。 The present invention relates to a yeast extract and a method for producing the same, and more particularly to a yeast extract excellent in kokumi imparting ability and masking ability and a method for producing the same.
近年調味料市場は消費者の本物志向に加えて健康志向も高まり、より自然な味、天然の風味を訴求した製品が求められている。単に旨味を付与するだけのグルタミン酸ソーダ（ＭＳＧ）やイノシン酸・グアニル酸といった単純な呈味性物質から、複雑な味を付与する複合調味料、ＨＶＰ（Hydrolyzed vegetable protein、植物性タンパク質加水分解物）、ＨＡＰ（Hydrolyzed animal protein、動物性タンパク質加水分解物）、各種エキス類の天然系調味料へと変遷を遂げてきている。 In recent years, the seasoning market has become more health-conscious in addition to consumers' real intentions, and there is a demand for products that appeal to more natural and natural flavors. A simple seasoning substance such as glutamic acid soda (MSG) or inosinic acid / guanylic acid that simply imparts umami, a complex seasoning that imparts a complex taste, HVP (Hydrolyzed vegetable protein, vegetable protein hydrolyzate) , HAP (Hydrolyzed animal protein, animal protein hydrolyzate) and various extracts have been transformed into natural seasonings.
現在、大きな潮流になりつつあるのが、濃厚感や後味の広がりを付与するコク味調味料である。食品にコク味を付与する物質としては、ペプチド、ピラジン類、フラン類、油脂・グリコーゲン・ゼラチン等の高分子物質や、オニオン・ガーリックなどの香味成分などが知られている。その中で、ペプチドは、味噌、納豆、チーズなどの発酵食品や食肉などの熟成食品に多く含まれ、コク味付与に重要な役割を果たしている。また、ペプチドは、アミノ酸等の味覚の発現時間が短い呈味成分に対して、その発現時間を伸ばす効果もあるので、味に厚みを持たせることができる。 Nowadays, a trend that is becoming a major trend is a rich-flavored seasoning that adds richness and aftertaste. Known substances that impart a rich taste to foods include peptides, pyrazines, furans, polymer substances such as fats, glycogen, and gelatin, and flavor components such as onion and garlic. Among them, peptides are often contained in fermented foods such as miso, natto and cheese, and aged foods such as meat, and play an important role in imparting rich taste. Moreover, since the peptide also has an effect of extending the expression time of a taste component such as an amino acid that has a short taste expression time, the taste can be made thicker.
例えば、発酵食品である魚醤油には、ペプチドが多く含まれ、食品にコク味を付与する作用がある。魚醤油は、日本では、しょっつる、いしるとして知られ、東南アジアでは、カンボジアのプラホック、ベトナムのニョクマム、タイのナンプラ等が有名である。
魚醤油のコク味には、製造工程の分解、熟成工程で魚介類のタンパク質が酵素により分解されてできるペプチド、アミノ酸が関与している（例えば特許文献１）。また、魚醤油の仕込み工程でアルカリ性プロテアーゼを添加し、ペプチド含有量を増加させコク味を増強することも開示されている（例えば特許文献２）。
For example, fish soy sauce, which is a fermented food, contains a large amount of peptides and has the effect of imparting a rich taste to the food. In Japan, fish soy sauce is known as “Sutsuru” and “Ishiru”, and in Southeast Asia, it is famous for Prahok in Cambodia, Nyokumam in Vietnam, Nampula in Thailand, and so on.
The richness of fish soy sauce involves peptides and amino acids formed by the degradation of fish and shellfish proteins by enzymes in the degradation and aging processes of the production process (for example, Patent Document 1). In addition, it is also disclosed that alkaline protease is added in the fish soy preparation process to increase the peptide content and enhance the rich taste (for example, Patent Document 2).
旨味調味料として、広く飲食品に利用されている酵母エキスにおいても、従来からある、旨味付与に加えて、ペプチドのコク味付与作用を利用したペプチド高含有の酵母エキスが開発されている（例えば、特許文献３、非特許文献１）。 As a umami seasoning, in yeast extract widely used for food and drink, in addition to the conventional umami imparting, a yeast extract having a high peptide content utilizing the richness imparting action of the peptide has been developed (for example, , Patent Document 3, Non-Patent Document 1).
しかし、従来のコク味付与酵母エキスでは、コク味と共に旨味などの味も付与されてしまう。そのため、添加する飲食品の味が変わることを望まない場合には、酵母エキスの添加量をごく少量に制限するか、添加することができなかった。 However, in the conventional kokumi-giving yeast extract, a taste such as umami is given along with the kokumi. Therefore, when it is not desired that the taste of the food or drink to be added changes, the addition amount of the yeast extract is limited to a very small amount or cannot be added.
一方、飲食品には、魚臭さや畜肉臭など不快な臭いを伴うものがあり、臭いをマスキングや低減する方法が報告されている。例えば、魚臭のマスキングに脂溶性ジンジャーフレーバーを添加したり（例えば特許文献４）、柑橘系香料とヨーグルト香料を添加すること（例えば特許文献５）が開示されている。また、畜肉臭のマスキングのためキノコ類の抽出物を添加する方法（例えば特許文献６）や、５’−イノシン酸、５’−グアニル酸を対固形分当り各々１〜５％、β−グルカン及びマンナンを対固形分当たり５％以上含有し、かつペプチド含量が対固形分当たり２０％以上である酵母エキスを添加する方法（例えば特許文献７）等が開示されている。 On the other hand, some foods and drinks have unpleasant odors such as fishy odor and livestock meat odor, and methods for masking or reducing the odor have been reported. For example, fat-soluble ginger flavor is added to masking fishy odor (for example, Patent Document 4), or citrus flavor and yogurt flavor are added (for example, Patent Document 5). In addition, a method of adding an extract of mushrooms for masking of livestock meat odor (for example, Patent Document 6), 5′-inosinic acid, 5′-guanylic acid 1 to 5% per solid, β-glucan And a method of adding a yeast extract containing 5% or more of mannan per solid content and a peptide content of 20% or more per solid content (for example, Patent Document 7).
しかし、これらのマスキング方法では、マスキングのために添加する物質自体が、味を有しているため、添加する食品の味を変えてしまい、その食品本来の味を変えたくない場合には、マスキング剤の添加量をごく少量に制限するか、添加することができなかった。 However, in these masking methods, since the substance added for masking itself has a taste, the taste of the food to be added is changed, and the original taste of the food is not desired to be changed. The addition amount of the agent was limited to a very small amount or could not be added.
本発明は、飲食品本来の味を損ねることなく、飲食品に濃厚感を付与する酵母エキスおよびその製法を提供することを課題とする。また本発明は、飲食品本来の味を損ねることなく、飲食品の不快な臭いをマスキング若しくは低減する酵母エキスおよびその製法を提供することを課題とする。 This invention makes it a subject to provide the yeast extract which provides a rich feeling to food-drinks, and its manufacturing method, without impairing the original taste of food-drinks. Moreover, this invention makes it a subject to provide the yeast extract which masks or reduces the unpleasant smell of food / beverage products, and its manufacturing method, without impairing the original taste of food / beverage products.
本発明者らは、かかる課題を解決すべく鋭意検討した結果、酵母エキス中のペプチド含量およびペプチド比率（全アミノ酸含量に対するペプチド含量の比率）を所定の数値以上に調製することにより、その酵母エキスが飲食品に濃厚感を与え、また飲食品の不快な臭いを低減あるいはマスキングする性質に優れることを見出した。また、この酵母エキスは遊離アミノ酸の含量も少なく、添加対象物となる飲食品本来の味を変化させにくいことも見出した。さらに、本発明者らは、５’−イノシン酸および５’−グアニル酸などのヌクレオチド類の含量を所定量以下に抑制することによって、飲食品本来の味質をより変化させにくい添加物とすることができる。 As a result of intensive studies to solve such problems, the present inventors have prepared the yeast extract by adjusting the peptide content and peptide ratio (ratio of peptide content to the total amino acid content) in the yeast extract to a predetermined value or more. Has been found to give a rich feeling to foods and drinks and to reduce or mask unpleasant odors of foods and drinks. In addition, the yeast extract has a low content of free amino acids, and it has been found that the original taste of foods and drinks to be added is difficult to change. Furthermore, the present inventors suppress the content of nucleotides such as 5′-inosinic acid and 5′-guanylic acid to a predetermined amount or less, thereby making an additive that hardly changes the original taste of food and drink. be able to.
上記本発明の酵母エキスを得るためには、予め酵母の菌体内酵素をすべて失活させた後、エンド型プロテアーゼを作用させることが好適である。このような方法を採用することにより、添加対象となる飲食品が元々持っているコク味を増強する作用を有するペプチド含量が高まり、遊離アミノ酸やヌクレオチド類の生成を抑制し、好ましい成分配合の酵母エキスを得られることを見出した。本発明は、係る知見に基づき完成されたものであり、具現化した態様として下記発明を提供するものである。 In order to obtain the yeast extract of the present invention, it is preferable to inactivate all the yeast intracellular enzymes in advance and then act on the endo-type protease. By adopting such a method, the content of the peptide having the effect of enhancing the richness originally possessed by the food and drink to be added is increased, the production of free amino acids and nucleotides is suppressed, and a yeast having a preferred component formulation It was found that an extract can be obtained. The present invention has been completed on the basis of such findings, and provides the following inventions as embodied embodiments.
〔１〕下記式（１）で示されるペプチドの含量が２０重量％以上であり、下記式（２）で示される全アミノ酸に対するペプチドの含有比率が８０％以上であり、５’−イノシン酸および５’−グアニル酸の合計含量が２重量％未満であり、かつ酵母エキスに含有される酸性ペプチドの含量が４重量％以上である酵母エキスであって、キャンディダ属又はサッカロマイセス属に属する酵母の菌体内酵素を失活させた後、菌体の細胞壁を破砕して得られる菌体処理物に、呈味性５’−ヌクレオチド類を生成させる酵素を添加せずに、エンド型プロテアーゼをアルカリ性領域で作用させて得られることを特徴とする酵母エキス。 [1] The content of the peptide represented by the following formula (1) is 20% by weight or more, the content ratio of the peptide with respect to all amino acids represented by the following formula (2) is 80% or more , 5′-inosinic acid and A yeast extract in which the total content of 5′-guanylic acid is less than 2% by weight and the content of acidic peptide contained in the yeast extract is 4% by weight or more , wherein the yeast extract belongs to the genus Candida or Saccharomyces. After inactivating the intracellular enzyme, the endo-protease is added to the alkaline region without the addition of an enzyme that generates a tasty 5′-nucleotide into the treated bacterial cell obtained by crushing the cell wall of the bacterial cell. Yeast extract characterized by being made to act on.
〔２〕上記〔１〕に記載の酵母エキスを含有することを特徴とする飲食品。
〔３〕上記〔１〕に記載の酵母エキスを含有することを特徴とする飲食品の不快臭低減剤
。
〔４〕酵母由来のペプチドを含有し、下記式（１）で示されるペプチドの含量が２０重量％以上であり、下記式（２）で示される全アミノ酸に対するペプチドの含有比率が８０％以上であり、５’−イノシン酸および５’−グアニル酸の合計含量が２重量％未満であり、かつ酵母エキスに含有される酸性ペプチドの含量が４重量％以上であって、キャンディダ属又はサッカロマイセス属に属する酵母の菌体内酵素を失活させた後、菌体の細胞壁を破砕して得られる菌体処理物に、呈味性５’−ヌクレオチド類を生成させる酵素を添加せずに、エンド型プロテアーゼをアルカリ性領域で作用させて得られる飲食品の不快臭低減剤。
[2] A food or drink comprising the yeast extract according to [1] above.
[3] An unpleasant odor reducing agent for foods and drinks comprising the yeast extract according to [1] above.
[4] It contains a peptide derived from yeast, the content of the peptide represented by the following formula (1) is 20% by weight or more, and the content ratio of the peptide to all amino acids represented by the following formula (2) is 80% or more. There, 5'total content of inosinic acid and 5'-guanylate is less than 2 wt%, and a is the content of the acidic peptide contained in the yeast extract 4% by weight or more, the genus Candida or Saccharomyces Endo type without adding an enzyme for producing a taste 5′-nucleotides to a treated cell obtained by crushing the cell wall of the yeast after inactivating the yeast intracellular enzyme belonging to An unpleasant odor reducing agent for food and drink obtained by allowing protease to act in an alkaline region.
〔５〕キャンディダ属又はサッカロマイセス属に属する酵母の菌体内酵素を失活させた後、菌体の細胞壁を破砕した菌体処理物に、呈味性５’−ヌクレオチド類を生成させる酵素を添加せずに、エンド型プロテアーゼをアルカリ性領域で作用させる、下記式（１）で示されるペプチドの含量が２０重量％以上であり、下記式（２）で示される全アミノ酸に対するペプチドの含有比率が８０％以上であり、５’−イノシン酸および５’−グアニル酸の合計含量が２重量％未満であり、かつ酵母エキスに含有される酸性ペプチドの含量が４重量％以上である酵母エキスの製造方法。
本発明によれば、飲食品本来の味の質を大きく変えることなく、飲食品に濃厚感を付与することができる。飲食品の濃厚感を付与または増強することにより、飲食品にコク味を付与もしくは増強することができる。さらに、味質の変化を必要としない又は望まない飲食品に濃厚感のみを付与する際に有効である。また、本発明によれば、コク味付与と共に、魚臭や畜肉臭などの飲食品の不快臭を低減あるいはマスキングすることができる。また、本発明の酵母エキス等は、安全性の高い天然素材の酵母から抽出しているものであるため、各種飲食品に広く安全に使用できる。 ADVANTAGE OF THE INVENTION According to this invention, a rich feeling can be provided to food / beverage products, without changing the quality of the original taste of food / beverage products. By imparting or enhancing the richness of the food / beverage product, it is possible to impart or enhance the richness of the food / beverage product. Furthermore, it is effective when only a rich feeling is imparted to a food or drink that does not require or do not require a change in taste quality. Moreover, according to this invention, unpleasant odor of food-drinks, such as fish odor and livestock meat odor, can be reduced or masked with rich taste provision. Moreover, since the yeast extract etc. of this invention are extracted from the highly safe natural material yeast, they can be widely used safely in various foods and drinks.
本発明の酵母エキスは、上記式（１）で示されるペプチドの含量が２０重量％以上であり、かつ上記式（２）で示される全アミノ酸に対するペプチドの含有比率が８０％以上に調製されたものである。本発明の酵母エキスは、このようなペプチド含量およびペプチド比率を有し、飲食品のコク味を増強する作用のあるペプチド含量が高いため、飲食品へのコク味増強効果、具体的には特に濃厚感などを付与する効果が高い。また、このようなペプチド含量およびペプチド比率を有する酵母エキスは、魚臭などの不快臭をマスキングする能力にも優れる。さらに、５’−グアニル酸、５’−イノシン酸等の呈味性５’−ヌクレオチド類やグルタミン酸ナトリウムなどの旨味成分となる遊離アミノ酸類の配合量が抑制されているため、添加対象物となる飲食品に旨味を加えないので、本来の味質を変化させにくい。 The yeast extract of the present invention was prepared such that the content of the peptide represented by the above formula (1) was 20% by weight or more, and the content ratio of the peptide with respect to all the amino acids represented by the above formula (2) was 80% or more. Is. Since the yeast extract of the present invention has such a peptide content and peptide ratio and has a high peptide content that has the effect of enhancing the rich taste of food and drink, the effect of enhancing the rich taste to food and drink, specifically, The effect of giving a rich feeling is high. Moreover, the yeast extract which has such a peptide content and a peptide ratio is excellent also in the capability to mask unpleasant odors, such as a fishy odor. Furthermore, since the blending amount of free amino acids serving as umami components such as 5′-guanylic acid, 5′-inosinic acid and the like and 5′-nucleotides and sodium glutamate is suppressed, it becomes an addition target. Because it does not add umami to foods and drinks, it is difficult to change the original taste.
グルタミン酸ナトリウムは旨味物質としてよく知られているが、本発明の酵母エキスにおいてはその含量は低い方が好ましく、具体的には好ましくは４重量％以下であり、より好ましくは２重量％以下である。 Sodium glutamate is well known as an umami substance, but in the yeast extract of the present invention, its content is preferably low, specifically 4 wt% or less, more preferably 2 wt% or less. .
なお、飲食品に味を付与する物質を呈味性物質ともいうが、本明細書において呈味性物質とは、塩味、甘味、旨味、苦み、辛み、酸味など、飲食品の味の質を規定することとなる主成分をいう。 A substance that imparts taste to a food or drink is also called a taste substance, but in this specification, a taste substance means the taste quality of a food or drink such as salty taste, sweet taste, umami, bitterness, hotness, and sourness. The main component that will be specified.
本明細書において、酵母エキスとは、酵母の細胞壁を破砕して得られる菌体内容物を主成分とする混合物のこという。細胞壁の破砕は、機械的・物理的な破砕および化学物質や酵素などによる化学的な破砕方法の双方が含まれる。酵母エキスの製法については下記にて詳説する。 In the present specification, the yeast extract refers to a mixture mainly composed of cell contents obtained by crushing cell walls of yeast. Cell wall crushing includes both mechanical and physical crushing and chemical crushing methods using chemical substances and enzymes. The method for producing the yeast extract will be described in detail below.
本明細書において、「濃厚感」とは、「味の強さ」と「持続性」を示す指標であり、例えば、飲食品を口にしたときに感じる「味の厚み」、「味の濃さ」、「味の持続性」等である。
また、「濃厚感」は、コク味を形成する要素の一つであり、コク味の強さを左右する。コク味とは、一般に食品の本格的な風味、複雑なおいしさを表す用語であり、甘味、塩味、酸味、苦味、辛味などの基本的な味質で、おいしさを表現できない場合に用いられ、具体的には、「持続感のある厚みのある強い旨味」といわれている。コク味は、「味の強さ」、「広がり」、「持続性」、「ハーモニー」などの要素から構成されており、味の「濃厚感」が強いと、コク味も強く感じることになる。
In the present specification, “thickness” is an index indicating “taste strength” and “sustainability”. For example, “taste thickness”, “taste strength” Sa ”,“ taste sustainability ”and the like.
Further, “richness” is one of the elements that form the rich taste, and affects the strength of the rich taste. Kokumi is a term that expresses the authentic flavor and complex taste of food in general, and is used when basic taste qualities such as sweet, salty, sour, bitter, and pungent cannot be expressed. Specifically, it is said to be “thick and strong umami with a lasting feeling”. The rich taste is made up of elements such as “strength of taste”, “spread”, “sustainability”, “harmony”, etc. If the “richness” of the taste is strong, the rich taste will also be felt strongly .
また、本明細書において、飲食品の不快臭とは、飲食品に含まれる不快な臭い全般を指し、例えば、魚臭、畜肉臭、大豆臭等が例示される。 Moreover, in this specification, the unpleasant odor of food / beverage products refers to the whole unpleasant odor contained in food / beverage products, for example, fish odor, livestock meat odor, soybean odor, etc. are illustrated.
本明細書においてペプチドとは、アミノ酸が２個〜数十個がペプチド結合を持って連結した化合物のことをいう。 In the present specification, a peptide refers to a compound in which 2 to several tens of amino acids are linked with a peptide bond.
本明細書においてペプチド含量とは、酵母エキスの絶乾重量に対するペプチドの重量含有率をいい、上記式１で算出される。 In this specification, the peptide content means the weight content of the peptide with respect to the absolute dry weight of the yeast extract, and is calculated by the above formula 1.
式１において、全アミノ酸重量とは、酵母エキス中に含まれる全アミノ酸重量のことを意味し、その値は、酵母エキス中のタンパク質を加水分解し、完全にアミノ酸に分解した後に、アミノ酸重量を測定し求める。遊離アミノ酸重量とは、酵母エキス中にもともとアミノ酸の形で含まれている遊離アミノ酸を意味し、その値は、酵母エキス中のタンパク質を加水分解せずにアミノ酸重量を測定し求める。 In Formula 1, the total amino acid weight means the total amino acid weight contained in the yeast extract, and the value is obtained by hydrolyzing the protein in the yeast extract and completely decomposing it into amino acids, and then calculating the amino acid weight. Measure and seek. The free amino acid weight means a free amino acid originally contained in the yeast extract in the form of an amino acid, and the value is obtained by measuring the amino acid weight without hydrolyzing the protein in the yeast extract.
本明細書においてペプチド比率とは、酵母エキス中の全アミノ酸含量に対するペプチド含量の割合を示したものであり、上記式２によって算出される。ペプチド比率が高いほど、酵母エキス中の全アミノ酸に対するペプチドの含有量は多い。 In the present specification, the peptide ratio indicates the ratio of the peptide content to the total amino acid content in the yeast extract, and is calculated by the above formula 2. The higher the peptide ratio, the higher the peptide content relative to all amino acids in the yeast extract.
各アミノ酸の含有量は、アミノ酸測定用ＨＰＬＣにより測定する。 The content of each amino acid is measured by HPLC for amino acid measurement.
分子量測定用ＨＰＬＣ（ゲル濾過クロマトグラフィー）によるペプチドの分子量測定より推察すると、濃厚感を増強するペプチドでは、分子量５００〜１００００の範囲のペプチドが大きな含有割合を占めているものと思われる。 As inferred from the molecular weight measurement of peptides by HPLC for molecular weight measurement (gel filtration chromatography), it is considered that peptides having a molecular weight in the range of 500 to 10,000 occupy a large content in peptides that enhance the richness.
濃厚感付与やマスキング効果を発揮させるためには、酵母エキスに含有される酸性ペプチドの含量は４重量％以上、より好ましくは５重量％以上であることが好適である。また、その分子量は、１０００〜５０００（アミノ酸１０個から５０個程度結合したもの）のものが、より好適である。 In order to exert a rich feeling and a masking effect, the content of the acidic peptide contained in the yeast extract is preferably 4% by weight or more, more preferably 5% by weight or more. Further, the molecular weight is more preferably 1000 to 5000 (a combination of about 10 to 50 amino acids).
酸性ペプチドとは、酸性アミノ酸であるアスパラギン酸、グルタミン酸などをペプチドの構成成分として多く含み、ペプチドの分子として中性領域下において酸性を呈するもの（負電荷を有するもの）をいう。酸性ペプチドとしては、カルボキシル基がリッチなペプチドが例示される。 An acidic peptide is one that contains many acidic amino acids aspartic acid, glutamic acid, and the like as constituents of the peptide and exhibits acidity (a negative charge) as a peptide molecule in a neutral region. Examples of acidic peptides include peptides rich in carboxyl groups.
本発明に記載の実施方法で、酵母エキスを製造すれば、原料となる酵母の種類によらず、酸性ペプチドの含有量は４重量％以上とすることができる。酸性ペプチドの含量は、本発明で用いるエンド型プロテアーゼによるところが大きいと考えられる。 If yeast extract is manufactured by the implementation method described in the present invention, the content of acidic peptide can be 4% by weight or more regardless of the type of yeast used as a raw material. It is considered that the content of the acidic peptide is largely due to the endo-type protease used in the present invention.
本発明において採用される酸性ペプチドは、酵母エキスの飲食品への添加量の１／１０程度という少量の添加量でも、濃厚感付与、マスキング効果が認められる。 The acidic peptide employed in the present invention can provide a rich feeling and a masking effect even with a small addition amount of about 1/10 of the addition amount of yeast extract to food and drink.
酸性ペプチドの分取は、酵母エキス水溶液を陰イオン交換樹脂に通し、吸着された酸性ペプチドを酢酸で溶出することにより行った。分子量の分画は、ゲル濾過クロマトグラフィーにより行うことができる。 The fractionation of the acidic peptide was performed by passing the aqueous yeast extract solution through an anion exchange resin and eluting the adsorbed acidic peptide with acetic acid. Molecular weight fractionation can be performed by gel filtration chromatography.
また、酵母エキス由来のペプチドによる濃厚感付与、マスキング作用は、ペプチドの構成アミノ酸の組成、組成比等によっても変化し得ると考えられる。 In addition, it is considered that the imparting of richness and the masking action by the peptide derived from yeast extract can also be changed by the composition, composition ratio, etc. of the constituent amino acids of the peptide.
本発明の酵母エキスが濃厚感付与およびマスキング作用に優れるのは、酵母エキス中の成分のうち、特にペプチドが、飲食品の元々持つコク味を増強する性質を有しているためであると考えられる。また、マスキング効果に関しては、主にペプチドがその効果を有していると推測されるが、酵母エキス中のその他の成分もペプチドと共存することにより、上記の効果を補強していると考えられる。 The reason why the yeast extract of the present invention is excellent in imparting richness and masking is considered to be because, among the components in the yeast extract, in particular, the peptide has the property of enhancing the richness of the food and drink. It is done. In addition, regarding the masking effect, it is presumed that the peptide mainly has the effect, but it is considered that other components in the yeast extract coexist with the peptide to reinforce the above effect. .
本発明の酵母エキスの好ましい形態としては、酵母エキス中において、酵母菌体内の核酸の分解により生成する５’−イノシン酸および５’−グアニル酸などの呈味性５’−ヌクレオチド類の含量を低く調製する。呈味性５’−ヌクレオチド類は旨味を付与するため、本発明の酵母エキスにおいてはそのような成分は低含有量であるこのが好ましい。そのようなヌクレオチド類として具体的には、５’−イノシン酸および５’−グアニル酸などが例示される。食品本来の味を変えずに濃厚感を付与するためには、５’−イノシン酸および５’−グアニル酸の含有量は、好ましくは２重量％未満、より好ましくは１重量％以下とする。 As a preferred form of the yeast extract of the present invention, the yeast extract contains the content of taste-dependent 5′-nucleotides such as 5′-inosinic acid and 5′-guanylic acid produced by the degradation of nucleic acids in the yeast cells. Prepare low. Since the tasty 5'-nucleotides impart umami, such components are preferably low in the yeast extract of the present invention. Specific examples of such nucleotides include 5'-inosinic acid and 5'-guanylic acid. In order to give a rich feeling without changing the original taste of food, the content of 5'-inosinic acid and 5'-guanylic acid is preferably less than 2% by weight, more preferably 1% by weight or less.
本発明の酵母エキスの好ましい一形態においては、酵母エキスのペプチド含量・比率が高く、かつ旨味を付与する成分である５’−イノシン酸、５’−グアニル酸の含量及びグルタミン酸ナトリウム含量が低いため、食品本来の味や香りを損ねることなく、コク味を付与する作用に極めて優れたエキスとすることができる。 In a preferred embodiment of the yeast extract of the present invention, the yeast extract has a high peptide content / ratio and a low content of 5′-inosinic acid, 5′-guanylic acid, and sodium glutamate, which are components imparting umami. It is possible to obtain an extract that is extremely excellent in the action of imparting a rich taste without impairing the original taste and aroma of food.
本発明における酵母エキスの原料となる酵母は、可食性のものであれば特に制限はなく、ビール酵母，パン酵母，アルコール酵母，清酒用酵母など一般に食品工業で用いられているものを使用することが出来る。このような酵母としては、例えばサッカロマイセス(Saccharomyces)属、ピキア(Pichia)属、ハンゼヌラ(Hansenula)属、デバリオマイセス(Debariomyces)属、キャンディダ(Candida)属、クリベロマイセス(Kluyveromyces)属、チゴサッカロマイセス(Zygosaccharomyces)属等に属する酵母が挙げられる。このような酵母の例としては、サッカロマイセス・セレビシエ(Saccharomyces cerevisiae IFO 1954, IFO 03019, IAM 4274)、ピキア・ステイピティス(Pichia stipitis IFO 1720)、ハンゼヌラ・アノマラ(Hansenula anomala IFO 1150)、デバリオマイセス・ハンセニイ(Debariomyces hansenii IFO 0855, IFO 1752)、キャンディダ・ユーティリス(Candida utilis IFO 0619, ATCC 15239)、クリベロマイセス・ラクチス(Kluyveromyces ATCC 56498)、クリベロマイセス・マルキシアヌス(Kluyveromyces marxianus ATCC 36534)、チゴサッカロマイセス・ルーキシー(Zygosaccharomyces rouxii ATCC 28253, ATCC 13356)等に属する酵母が挙げられる。 The yeast used as a raw material for the yeast extract in the present invention is not particularly limited as long as it is edible, and those commonly used in the food industry such as brewer's yeast, baker's yeast, alcoholic yeast, and sake yeast are used. I can do it. Examples of such yeast include the genus Saccharomyces, the genus Pichia, the genus Hansenula, the genus Debariomyces, the genus Candida, the genus Kluyveromyces, the genus Zygosaccharomies Examples include yeast belonging to the genus and the like. Examples of such yeasts include Saccharomyces cerevisiae (IFO 1954, IFO 03019, IAM 4274), Pichia stipitis IFO 1720, Hansenula anomala IFO 1150, Debariomyces hansenii IFO 0855, IFO 1752) Candida utilis IFO 0619, ATCC 15239 28253, ATCC 13356) and the like.
これらの酵母の中から１種若しくは２種以上の酵母を用いる。中でもキャンディダ属酵母やサッカロマイセス属に属する酵母は菌体収率が高く培養もしやすいので、工業的生産を行う上で好適である。なお、酵母の培養方法に特に制限はなく、定法に従って行うことができる。 One or more yeasts are used from these yeasts. Among them, the yeast belonging to the genus Candida and the genus Saccharomyces is suitable for industrial production because of its high cell yield and easy culturing. In addition, there is no restriction | limiting in particular in the culture | cultivation method of yeast, It can carry out according to a usual method.
以下に、本発明の酵母エキスの好ましい製法について詳説する。
本発明の酵母エキスは、酵母菌体の細胞壁を破砕した菌体処理物にエンド型プロテアーゼを作用させることによって得ることができる。エンド型プロテアーゼを使用することで、グルタミン酸およびその塩類など呈味性物質の遊離アミノ酸の生成量を抑えるとともに、ペプチド含量を高め、上記本発明の酵母エキスを得ることができる。また、より好ましい形態としては、酵母の菌体内酵素を失活させた後、菌体の細胞壁を破砕した菌体処理物にエンド型プロテアーゼを作用させる。このような工程を取ることにより、酵母に含まれる核酸の分解によって、呈味性５’−ヌクレオチド類が生成することを抑制し、これらの含量が低い酵母エキスを簡便に得ることができる。
Below, the preferable manufacturing method of the yeast extract of this invention is explained in full detail.
The yeast extract of the present invention can be obtained by allowing an endo-type protease to act on a treated product obtained by disrupting the cell walls of yeast cells. By using an endo-type protease, it is possible to suppress the production amount of free amino acids of taste substances such as glutamic acid and salts thereof, and to increase the peptide content, thereby obtaining the yeast extract of the present invention. Moreover, as a more preferable form, after inactivating the yeast cell enzyme, an endo-type protease is allowed to act on the treated cell product obtained by crushing the cell wall of the cell. By taking such a step, it is possible to suppress the production of taste-preferring 5′-nucleotides due to the degradation of the nucleic acid contained in the yeast, and to easily obtain a yeast extract having a low content thereof.
菌体の細胞壁を破砕し菌体処理物を得る方法は、細胞壁を破壊することができれば特に限定はなく、例えば、自己消化法、酵素分解法、酸分解法、超音波破砕法、ホモジナイザー法、圧力破砕法、ビーズ衝撃法、摩砕法、凍結融解法などが例示され、好ましくは酵素を添加して細胞壁を破壊する酵素分解法などが例示される。 There is no particular limitation on the method for obtaining a cell-treated product by crushing the cell wall of the cell, as long as the cell wall can be destroyed. For example, the autolysis method, the enzyme decomposition method, the acid decomposition method, the ultrasonic crushing method, the homogenizer method, Examples thereof include a pressure crushing method, a bead impact method, a crushing method, a freeze-thawing method, and the like, and preferably an enzyme decomposition method in which an enzyme is added to break the cell wall.
酵素分解法による製法を具体例として説明する。本発明の酵母エキスの製造に適した酵素分解法の一例としては、（ｉ）加熱などにより菌体内酵素を失活させる工程、（ii）酵母またはその菌体を含む培養物等に細胞壁溶解酵素を添加して菌体の細胞壁を分解し菌体処理物を得る工程、（iii）菌体内タンパク質をペプチドに分解するエンド型プロアーゼを添加する工程の３工程を含む。上記の工程中、エンド型プロテアーゼを添加する前に菌体内酵素を失活させる工程を行うこと以外は、順序は任意である。より好ましくは、上記（ｉ）の工程以降は、酵母又は菌体処理物に呈味性５’−ヌクレオチド類を生成させないようにする。具体的には、酵母内のリボ核酸を加水分解する５’−ホスホジエステラーゼ、および５’−アデニル酸から５’−イノシン酸を生成させる５’−アデニル酸デアミナーゼを作用させないようにすることが望ましい。本発明の酵母エキスの製造として好適な酵素分解法は、呈味性物質である５’−ヌクレオチド類を生成させないため、核酸を分解しないこと並びに、ペプチド含量を高めると共に呈味性物質であるグルタミン酸ナトリウムなどの遊離のアミノ酸含量を少なくすることができる。 A production method using an enzymatic decomposition method will be described as a specific example. As an example of the enzymatic degradation method suitable for the production of the yeast extract of the present invention, (i) a step of inactivating the intracellular enzyme by heating or the like, (ii) a cell wall lytic enzyme in a yeast or a culture containing the bacterial cell, etc. Is added to decompose the cell wall of the cells to obtain a treated product of the cells, and (iii) a step of adding endo-prose that decomposes the proteins in the cells into peptides. In the above steps, the order is arbitrary except that a step of inactivating the intracellular enzyme is performed before the endo-type protease is added. More preferably, after the step (i), the yeast or the treated bacterial cell product is prevented from producing taste-preferring 5'-nucleotides. Specifically, it is desirable not to act on 5'-phosphodiesterase that hydrolyzes ribonucleic acid in yeast and 5'-adenylate deaminase that generates 5'-inosinic acid from 5'-adenylic acid. The enzymatic degradation method suitable for production of the yeast extract of the present invention does not generate 5′-nucleotides that are tasty substances, so that it does not degrade nucleic acids, and increases the peptide content and glutamic acid that is a tasty substance. The content of free amino acids such as sodium can be reduced.
酵素の失活は、例えば加熱により行うことができる。酵母を１０−１５％程度の適当な濃度で水に懸濁させたのち、８０−１２０℃好ましくは９０−１００℃で加熱し、菌体内酵素の失活を行う。加熱時間は１０分程度で十分である。 The enzyme can be deactivated by heating, for example. The yeast is suspended in water at an appropriate concentration of about 10-15%, and then heated at 80-120 ° C., preferably 90-100 ° C., to inactivate intracellular enzymes. A heating time of about 10 minutes is sufficient.
上記（ii）の工程においては、細胞壁溶解酵素を作用させ、菌体の細胞壁を破砕し、細胞内の成分を溶出させる。酵素添加量、酵素反応温度、ｐＨは特に限定するものではなく、各々の酵素の最適条件下で行えばよい。細胞壁溶解酵素としては、酵母の細胞壁は、β−１，３−グルカンとマンナンが主成分であるので、グルカナーゼ、マンナナーゼを含有し、酵母細胞壁を溶解するに十分な活性を有するものであればよい。例えば市販の細胞壁溶解酵素としては、ＹＬ−１５（天野エンザイム（株）製）、ツニカーゼ（大和化成（株）製），キタラーゼ（クミアイ化学（株）製）などがあげられる。 In the step (ii), a cell wall lytic enzyme is allowed to act so as to crush the cell wall of the microbial cells and to elute intracellular components. The amount of enzyme added, enzyme reaction temperature, and pH are not particularly limited, and may be performed under the optimum conditions for each enzyme. As the cell wall lytic enzyme, β-1,3-glucan and mannan are the main components of the yeast cell wall, so long as they contain glucanase and mannanase and have sufficient activity to lyse the yeast cell wall. . Examples of commercially available cell wall lytic enzymes include YL-15 (Amano Enzyme Co., Ltd.), Tunicase (Daiwa Kasei Co., Ltd.), Kitalase (Kumiai Chemical Co., Ltd.) and the like.
また、上記（iii）の工程においては、菌体に含まれていたタンパク質を分解して低分子量化し、ペプチドとする。酵素添加量、酵素反応温度は特に限定するものではなく、各々の酵素の最適条件下で行えばよい。ｐＨについても特に限定するものではないが、アルカリ性領域で作用させるほうが、それ以外の領域で作用させるよりもペプチド含量、比率を高くすることができ、好適である。プロテアーゼには、ポリペプチド鎖の末端からペプチド結合を切断し、アミノ酸を１個ずつ遊離させるエキソ型と、ポリペプチド鎖の中間からペプチド結合を切断するエンド型があるが、本発明においては、ペプチド含量を高めるため、エンド型主体のものを使用する。エンド型プロテアーゼはその純度の高いものが好ましいが、実際の製造の場面では不可避的に不純物を含み得る。市販のプロテアーゼを使用する際は、本発明のペプチド含量及びペプチド比率を満たす酵母エキスを得られるものであれば構わないが、エンド型プロテアーゼの純度が８割以上、より好ましくは９割以上のものが好ましい。市販のエンド型プロテアーゼ製剤としては、例えば、アマノＮ（天野エンザイム（株）製）、プロチンＦＮ（大和化成（株）製）等があげられる。 In the step (iii), the protein contained in the microbial cells is decomposed to lower the molecular weight to obtain a peptide. The amount of enzyme added and the enzyme reaction temperature are not particularly limited, and may be performed under the optimum conditions for each enzyme. The pH is not particularly limited, but the action in the alkaline region is more preferable because the peptide content and ratio can be increased than in the other regions. Proteases include exo-types that cleave peptide bonds from the ends of polypeptide chains and release amino acids one by one, and endo-types that cleave peptide bonds from the middle of polypeptide chains. In order to increase the content, the end type main body is used. An endo-type protease having a high purity is preferred, but it may inevitably contain impurities in actual production situations. When using a commercially available protease, any yeast extract that satisfies the peptide content and peptide ratio of the present invention may be used, but the purity of the endo-type protease is 80% or more, more preferably 90% or more. Is preferred. Examples of commercially available endo-type protease preparations include Amano N (manufactured by Amano Enzyme Co., Ltd.), protin FN (manufactured by Daiwa Kasei Co., Ltd.), and the like.
上記工程（ii）と工程（iii）とは同時に行ってもよいし、また工程（ii）を行ってから、工程（iii）を行ってもよい。 The step (ii) and the step (iii) may be performed at the same time, or the step (iii) may be performed after performing the step (ii).
各酵素反応後、遠心分離して上澄液を回収し、必要に応じて濃縮し、スプレードライ等の方法により乾燥させ、固形物として酵母エキスを得ることができる。また、高濃度に濃縮し、液状濃縮エキスとしても良い。 After each enzyme reaction, the supernatant is collected by centrifugation, concentrated as necessary, and dried by a method such as spray drying to obtain a yeast extract as a solid. Moreover, it concentrates to high concentration and is good also as a liquid concentrated extract.
本発明における酵母エキスは、各種飲食品全般に用いることができる。各種飲食品としては、調理済み食品、水産加工品、畜肉加工品、漬け物、佃煮、健康食品、乳製品、スナック食品、菓子、冷菓子、スープ、調理用シーズニング、タレ、麺つゆ、ドレッシング、ソース、ケチャップ等の調味料、カレー、キムチなどの辛みを有する食品、豆乳、その他の飲料等が挙げられる。 The yeast extract in the present invention can be used for various foods and beverages in general. Various foods and beverages include cooked foods, processed fishery products, processed meat products, pickles, boiled foods, health foods, dairy products, snack foods, confectionery, cold confectionery, soup, seasoning for cooking, sauce, noodle soup, dressing, sauce And seasonings such as ketchup, spicy foods such as curry and kimchi, soy milk, and other beverages.
これらの飲食品に本発明の酵母エキスを添加することにより、濃厚感を付与し、コク味を増強することができる。中でも、塩性食品に対しては、塩辛みをまろやかにすることができ、辛み食品に対しては、辛みを弱めずにまろやかさと濃厚感を付与することができる。 By adding the yeast extract of this invention to these food-drinks, a rich feeling can be provided and richness can be reinforced. Among them, salty food can be mellowed for salty foods, and mellowness and richness can be imparted to spicy foods without reducing the hotness.
本発明の酵母エキスは、旨味成分の含有量が低いため、特に旨味を必要としない或いは味を変化させることを望まない飲食品について、濃厚感のみを付与することに特に有用である。 Since the content of the umami component is low, the yeast extract of the present invention is particularly useful for imparting only a rich feeling to foods and drinks that do not require umami or change the taste.
また、飲食品の不快臭もマスキング又は低減することができるが、特に、水産加工品の魚臭、畜肉加工品の獣臭、豆乳、豆腐等の大豆タンパク臭などのマスキング効果が優れている。 Moreover, although the unpleasant odor of food / beverage products can also be masked or reduced, the masking effect of fish odor of processed fishery products, animal odor of processed meat products, soybean protein odor such as soy milk, tofu, etc. is particularly excellent.
本発明の酵母エキスの飲食品への添加量は、濃厚感を付与するためには、食品の種類により異なるが、一般に食品に対し、酵母エキスとして０．０５重量％以上であれば充分な効果を得ることが可能である。また、味の濃い食品への添加については、少し多めに添加した方がよい。上限は特に制限はないが、経済性及び添加効果を考慮すると５重量％以下が好ましい。従って、食品に対し、酵母エキスとして０．０５重量％〜５重量％添加することが好ましい。さらに望ましくは０．１重量％〜２重量％添加することが好ましい。 The amount of the yeast extract of the present invention added to foods and drinks varies depending on the type of food to give a rich feeling, but generally 0.05% by weight or more as a yeast extract is sufficient for food. It is possible to obtain In addition, it is better to add a little more for a food with a strong taste. The upper limit is not particularly limited, but is preferably 5% by weight or less in consideration of economic efficiency and addition effect. Therefore, it is preferable to add 0.05% by weight to 5% by weight as a yeast extract to food. More desirably, 0.1 to 2% by weight is preferably added.
また、本発明の酵母エキスおよび酵母エキスに由来するペプチド製剤は、飲食品用の不快臭低減剤とすることができる。すなわち、本発明の不快臭低減剤は、上記酵母エキスをそのままでもよいし、また、酵母エキスに精製等の処理を施して、上記式（１）で示されるペプチドの含量が２０重量％以上であり、かつ上記式（２）で示される全アミノ酸に対するペプチドの含有比率が８０％以上に調製されたペプチド組成物に加工したものであってもよい。 Moreover, the peptide formulation derived from the yeast extract and yeast extract of this invention can be used as the unpleasant odor reducing agent for food-drinks. That is, the unpleasant odor reducing agent of the present invention may be the yeast extract as it is, or the yeast extract is subjected to a treatment such as purification, so that the content of the peptide represented by the formula (1) is 20% by weight or more. Yes, and may be processed into a peptide composition prepared such that the content ratio of peptides to all amino acids represented by the above formula (2) is 80% or more.
本発明における飲食品の不快臭低減剤として、本発明の酵母エキスの飲食品への添加量は、添加する食品の種類により調整されるが、一般に飲食品に対し、酵母エキスとして０．０５重量％以上であれば充分な効果を得ることが可能である。酵母エキスの添加量の上限は特に制限はないが、経済性及び添加効果を考慮すると５重量％以下が好ましい。従って、本発明の酵母エキスがその効果を充分に発揮するためには、食品に対し、酵母エキスとして０．０５重量％〜５重量％添加することが好ましい。さらに望ましくは０．１重量％〜２重量％添加することが好ましい。 As an unpleasant odor reducing agent for food and drink in the present invention, the amount of the yeast extract of the present invention added to the food and drink is adjusted depending on the type of food to be added. If it is at least%, a sufficient effect can be obtained. The upper limit of the amount of yeast extract added is not particularly limited, but is preferably 5% by weight or less in view of economy and the effect of addition. Therefore, in order for the yeast extract of this invention to fully exhibit the effect, it is preferable to add 0.05 to 5 weight% as a yeast extract with respect to a foodstuff. More desirably, 0.1 to 2% by weight is preferably added.
また、不快臭低減剤としてのペプチドの食品への添加量は、添加する食品の種類に調整されるが、一般に食品に対し、０．０１重量％以上であれば十分な効果を得ることが可能である。ペプチドの添加量の上限は特に制限はないが、経済性及び添加効果を考慮すると１重量％以下が好ましい。従って、本発明のペプチドがその効果を充分に発揮するためには、食品に対し、０．０１重量％〜１重量％添加することが好ましい。さらに望ましくは０．０２重量％〜０．４重量％添加することが好ましい。 In addition, the amount of peptide added to food as an unpleasant odor reducing agent is adjusted to the type of food to be added, but in general, sufficient effects can be obtained if it is 0.01% by weight or more with respect to food. It is. The upper limit of the amount of peptide to be added is not particularly limited, but is preferably 1% by weight or less in consideration of economy and the effect of addition. Therefore, in order for the peptide of the present invention to exert its effect sufficiently, it is preferable to add 0.01% by weight to 1% by weight to the food. More desirably, 0.02 wt% to 0.4 wt% is added.
本発明の酵母エキスは、添加する飲食品に求められる性質に応じて、他の呈味成分や各種調味料などと併用することも可能である。 The yeast extract of the present invention can be used in combination with other taste ingredients and various seasonings depending on the properties required for the food or drink to be added.
以下、本発明を実施例により詳細に説明するが、本発明はこれに限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to this.
本実施例中の％は、ペプチド比率を除き重量％を意味する。
［遊離アミノ酸及び全アミノ酸の測定方法］
酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、高速液体クロマトグラフィーシステム（東ソー（株）製）を用い、以下の条件で定量した。
カラム：ＴＳＫ−ＧＥＬ Ａｍｉｎｏ Ｐａｋ（東ソー社製）
検出：蛍光検出
溶離液：クエン酸バッファーによるグラジエント溶出（ｐＨ３→ｐＨ９）
In the examples,% means% by weight excluding the peptide ratio .
[Measurement method of free amino acids and total amino acids]
The free amino acids and total amino acid contents in the yeast extract were quantified under the following conditions using a high performance liquid chromatography system (manufactured by Tosoh Corporation).
Column: TSK-GEL Amino Pak (manufactured by Tosoh Corporation)
Detection: Fluorescence detection Eluent: Gradient elution with citrate buffer (pH 3 → pH 9)
［イノシン酸およびグアニル酸の測定方法］
酵母エキス中のイノシン酸およびグアニル酸含量は、高速液体クロマトグラフィーシステム（島津製作所製）を用い、以下の条件で定量した。
カラム：Ｓｈｏｄｅｘ Ａｓａｈｉｐａｋ ＧＳ−３２０，７Ｅ （昭和電工社製）
検出：ＵＶ２６０ｎｍ
溶離液：１０ｍＭリン酸ナトリウム
[Method for measuring inosinic acid and guanylic acid]
The inosinic acid and guanylic acid contents in the yeast extract were quantified under the following conditions using a high performance liquid chromatography system (manufactured by Shimadzu Corporation).
Column: Shodex Asahipak GS-320, 7E (manufactured by Showa Denko)
Detection: UV260nm
Eluent: 10 mM sodium phosphate
［グルタミン酸ナトリウムの測定方法］
酵母エキス中のグルタミン酸ナトリウムの測定は、市販のグルタミン酸測定キット（グルタミン酸 Ｆ−キット：発売元Ｊ．Ｋ．インターナショナル社）を用いて定量した。
[Measurement method of sodium glutamate]
The measurement of sodium glutamate in the yeast extract was quantified using a commercially available glutamic acid measurement kit (glutamic acid F-kit: distributor JK International).
［実施例１］酵母エキス１の調製
サッカロマイセス・セレビシエ(IFO 1954)を、糖蜜１ｍ3に対し、リン酸一アンモニウム３．３ｋｇ、塩化アンモニウム６．８ｋｇ、塩化カリウム１．７ｋｇ、硫酸マグネシウム１．７ｋｇ、硫酸亜鉛１７ｇ、微量金属類（ホウ酸０．１５ｇ、硫酸マンガン０．０７５ｇ、硫酸銅０．１０ｇ、塩化第二鉄０．６２ｇ、モリブデン酸ナトリウム０．０６３ｇ）を加えた培地（以下、糖蜜培地という）を用いて約１日間（２０時間から２４時間）培養し、集菌洗浄後、それを水に懸濁し、酵母スラリー（菌体濃度１５％）１０００ｍｌを調製した。
[Example 1] Preparation of yeast extract 1 Saccharomyces cerevisiae (IFO 1954) was prepared by adding 3.3 kg of monoammonium phosphate, 6.8 kg of ammonium chloride, 1.7 kg of potassium chloride, and 1.7 kg of magnesium sulfate to 1 m 3 of molasses. , Medium supplemented with 17 g of zinc sulfate and trace metals (boric acid 0.15 g, manganese sulfate 0.075 g, copper sulfate 0.10 g, ferric chloride 0.62 g, sodium molybdate 0.063 g) The culture medium was used for about 1 day (20 to 24 hours), and after collecting and washing the cells, it was suspended in water to prepare 1000 ml of a yeast slurry (cell concentration 15%).
酵母スラリーを、９５℃、１０分加熱し菌体内酵素を失活させた後、ｐＨを６に調製し、細胞壁溶解酵素（商品名：ＹＬ−１５（天野エンザイム（株）製）を１.５ｇ添加し５０℃にて４時間反応させた。 The yeast slurry was heated at 95 ° C. for 10 minutes to inactivate intracellular enzymes, adjusted to pH 6 and 1.5 g of cell wall lytic enzyme (trade name: YL-15 (manufactured by Amano Enzyme)). The mixture was added and reacted at 50 ° C. for 4 hours.
その後エンド型主体のプロテアーゼ（商品名：「アマノＮ」（天野エンザイム（株）製）を１ｇ添加しｐＨ５に調製後７時間反応させた。
反応後、遠心分離により上澄み液を回収し、粉末乾燥し、１１０ｇの酵母エキス１を得た。
Thereafter, 1 g of an endo-type protease (trade name: “Amano N” (manufactured by Amano Enzyme Co., Ltd.) was added to adjust to pH 5 and reacted for 7 hours.
After the reaction, the supernatant was collected by centrifugation and dried by powder to obtain 110 g of yeast extract 1.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々６．７％、３５．０％であり、ペプチド含量は２８．３％、ペプチド比率は８３．９％であった。 The free amino acids and total amino acids in the yeast extract were 6.7% and 35.0%, the peptide content was 28.3%, and the peptide ratio was 83.9%.
［実施例２］酵母エキス２の調製
キャンディダ・ユーティリス(IFO 0619)を、糖蜜培地を用いて培養し、集菌洗浄後、それを水に懸濁し、酵母スラリー（菌体濃度１５％）１０００ｍｌを調製した。
[Example 2] Preparation of yeast extract 2 Candida utilis (IFO 0619) was cultivated using molasses medium, collected and washed, suspended in water, and yeast slurry (cell concentration 15%) 1000 ml was prepared.
酵母スラリーを９５℃、１０分加熱し菌体内酵素を失活させた後ｐＨを６.５に調製した後、細胞壁溶解酵素（商品名：ツニカーゼ（大和化成（株）製）を１.８ｇ添加し４０℃にて５時間反応させた。 After the yeast slurry was heated at 95 ° C. for 10 minutes to inactivate intracellular enzymes, the pH was adjusted to 6.5, and 1.8 g of cell wall lytic enzyme (trade name: Tunicase (manufactured by Daiwa Kasei Co., Ltd.) was added. And reacted at 40 ° C. for 5 hours.
その後、エンド型プロテアーゼ（商品名：「アマノＰ」（天野エンザイム（株）製）を１ｇ添加し、ｐＨ６に調製後、５時間反応させた。反応後、遠心分離により上澄み液を回収し、粉末乾燥し、１０８ｇの酵母エキス２を得た。 Thereafter, 1 g of endo-type protease (trade name: “Amano P” (manufactured by Amano Enzyme Co., Ltd.) was added, and the mixture was reacted for 5 hours after adjusting to pH 6. After the reaction, the supernatant was collected by centrifugation and powdered. Dried to obtain 108 g of yeast extract 2.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々６．３％、３９．０％であり、ペプチド含量は３２．７％、ペプチド比率は８６．０％であった。 The free amino acids and total amino acids in the yeast extract were 6.3% and 39.0%, the peptide content was 32.7%, and the peptide ratio was 86.0%.
［実施例３］酵母エキス３の調製
実施例２の酵母スラリーを、９５℃、１０分加熱し菌体内酵素を失活させた後ｐＨを９.５に調製した後、細胞壁溶解酵素（商品名：ＹＬ−１５（天野エンザイム（株）製）を１.５ｇ、及びエンド型プロテアーゼ（商品名：「プロレーザー」（天野エンザイム（株）製）を０．５ｇ添加し５０℃にて４時間反応させた。
[Example 3] Preparation of yeast extract 3 The yeast slurry of Example 2 was heated at 95 ° C for 10 minutes to inactivate intracellular enzymes and then adjusted to pH 9.5. : 1.5 g of YL-15 (manufactured by Amano Enzyme) and 0.5 g of endo-type protease (trade name: “Prolaser” (manufactured by Amano Enzyme)) were added and reacted at 50 ° C. for 4 hours. I let you.
反応後、遠心分離し上澄み液を回収し、粉末乾燥し、１１０ｇの酵母エキス３を得た。
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々４．５％、４５．０％であり、ペプチド含量は４０．５％、ペプチド比率は９０．０％であった。
After the reaction, the mixture was centrifuged and the supernatant was collected and dried to obtain 110 g of yeast extract 3.
The free amino acid content and total amino acid content in this yeast extract were 4.5% and 45.0%, respectively, the peptide content was 40.5%, and the peptide ratio was 90.0%.
［比較例１］酵母エキス４の調製
上記実施例１において調製した酵母スラリーを、９５℃、１０分間加熱し、菌体内酵素を失活させた後、細胞壁溶解酵素（商品名：ＹＬ−１５（天野エンザイム（株）製））及びエキソ型主体のプロテアーゼ（商品名：プロチンＦＮ（大和化成（株）製））を同時に添加し、約１２時間、５０℃、ｐＨ６で反応させた。反応後、遠心分離し上澄み液を回収し粉末乾燥し、酵母エキス４を得た。
Comparative Example 1 Preparation of Yeast Extract 4 The yeast slurry prepared in Example 1 was heated at 95 ° C. for 10 minutes to inactivate intracellular enzymes, and then cell wall lytic enzyme (trade name: YL-15 ( Amano Enzyme Co., Ltd.)) and exo-type protease (trade name: Protin FN (manufactured by Yamato Kasei Co., Ltd.)) were added simultaneously and allowed to react for about 12 hours at 50 ° C. and pH 6. After the reaction, the mixture was centrifuged and the supernatant was collected and dried to obtain yeast extract 4.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々１５．２％、４２．５％であり、ペプチド含量は２７．３％、ペプチド比率は６４．２％であった。 The free amino acid content and total amino acid content in this yeast extract were 15.2% and 42.5%, respectively, the peptide content was 27.3%, and the peptide ratio was 64.2%.
［比較例２］酵母エキス５の調製
上記実施例２において調製したの酵母スラリーを、９５℃、１０分間加熱し、菌体内酵素を失活させた後、細胞壁溶解酵素（商品名：ＹＬ−１５（天野エンザイム（株）製））及びエキソ型主体のプロテアーゼ（商品名：プロチンＦＮ（大和化成（株）製））を同時に添加し、約１２時間、５０℃、ｐＨ６で反応させた。、反応後、遠心分離し上澄み液を回収し粉末乾燥し、酵母エキス５を得た。
[Comparative Example 2] Preparation of yeast extract 5 The yeast slurry prepared in Example 2 was heated at 95 ° C for 10 minutes to inactivate intracellular enzymes, and then cell wall lytic enzyme (trade name: YL-15). (Amano Enzyme Co., Ltd.) and exo-type protease (trade name: Protin FN (Daiwa Kasei Co., Ltd.)) were added simultaneously and allowed to react for about 12 hours at 50 ° C. and pH 6. After the reaction, the mixture was centrifuged and the supernatant was collected and dried to obtain yeast extract 5.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々１６．４％、４３．５％であり、ペプチド含量は２７．１％、ペプチド比率は６２．３％であった。 The free amino acid content and total amino acid content in this yeast extract were 16.4% and 43.5%, respectively, the peptide content was 27.1%, and the peptide ratio was 62.3%.
［比較例３］酵母エキス６の調製
上記実施例１において調製した酵母スラリーを、ｐＨ６、４０℃で２０時間保持することで、自己消化法により分解した。反応後、反応後、遠心分離し上澄み液を回収し粉末乾燥し、酵母エキス６を得た。
[Comparative Example 3] Preparation of yeast extract 6 The yeast slurry prepared in Example 1 was maintained at pH 6 and 40 ° C for 20 hours, and decomposed by the self-digestion method. After the reaction, the reaction was centrifuged, and the supernatant was collected and dried to obtain yeast extract 6.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々３０．４％、５０．２％であり、ペプチド含量は１９．８％、ペプチド比率は３９．４％であった。 The free amino acid content and total amino acid content in this yeast extract were 30.4% and 50.2%, respectively, the peptide content was 19.8%, and the peptide ratio was 39.4%.
［比較例４］酵母エキス７の調製
実施例２において調製した酵母スラリーを、ｐＨ６、４０℃で２０時間保持することで、自己消化法により分解した。反応後、反応後、遠心分離し上澄み液を回収し粉末乾燥し、酵母エキス７を得た。
[Comparative Example 4] Preparation of yeast extract 7 The yeast slurry prepared in Example 2 was decomposed by the self-digestion method by keeping it at pH 6 and 40 ° C for 20 hours. After the reaction, after the reaction, the mixture was centrifuged and the supernatant was collected and dried to obtain yeast extract 7.
この酵母エキス中の遊離アミノ酸及び全アミノ酸含量は、各々３１．５％、５２．３％であり、ペプチド含量は２０．８％、ペプチド比率は３９．８％であった。
実施例１〜３として調製した酵母エキス並びに比較例１〜４として調製した酵母エキス１〜７および市販の酵母エキスの成分組成を表１にまとめた。
The free amino acids and total amino acids in the yeast extract were 31.5% and 52.3%, the peptide content was 20.8%, and the peptide ratio was 39.8%.
The component composition of the yeast extract prepared as Examples 1-3, the yeast extracts 1-7 prepared as Comparative Examples 1-4, and a commercially available yeast extract was put together in Table 1.
コク味、特に濃厚感についての各官能試験は、以下の方法で行った。
１０名のパネラーにより、酵母エキス無添加のものを比較対照として、コク味および濃厚感を４段階（強い３点、弱い２点、極めて弱い１点、なし０点）で、その評価を行い、１０名の評価の平均値を求め、前記４段階にそって３以上を「強い」、２以上３未満を「弱い」、１以上２未満を「極めて弱い」、０以上１未満を「なし」とした。官能試験の結果とした。また、味の変化についてもその傾向を評価した。
Each sensory test for the body taste, particularly the rich feeling was performed by the following method.
10 panelists evaluated the taste and richness in 4 levels (strong 3 points, weak 2 points, extremely weak 1 point,
［官能試験１］
市販のコンソメスープ１００ｍｌに対し、酵母エキス１を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 1]
0.1 g of yeast extract 1 was added to 100 ml of commercially available consomme soup and kept at 60 ° C., and a sensory test was performed.
［官能試験２］
市販のだし１００ｍｌに対し、酵母エキス２を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 2]
0.1 g of yeast extract 2 was added to 100 ml of commercially available stock and kept at 60 ° C., and a sensory test was performed.
［官能試験３］
市販のだし１００ｍｌに対し、酵母エキス３を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 3]
0.1 g of yeast extract 3 was added to 100 ml of commercially available stock and kept at 60 ° C., and a sensory test was performed.
［官能試験４］
下記の処方にて常法により麺つゆを作った。麺つゆ１００ｍｌに対し、酵母エキス１を０．１ｇ添加し、官能試験を行った。
[Sensory test 4]
Noodle soup was made by the conventional method with the following formulation. 0.1 g of yeast extract 1 was added to 100 ml of noodle soup, and a sensory test was conducted.
麺つゆの処方 １０００ｇ中
醤油 ５０ｇ
砂糖 ３５ｇ
食塩 ５ｇ
鰹だし ９１０ｍｌ
官能試験１から４の結果を表２に示す。
Soy sauce 50g in 1000g noodle soup
35g sugar
5g salt
910ml dashi stock
The results of sensory tests 1 to 4 are shown in Table 2.
表２の結果から、本発明品は、単味であり味の変化がわかりやすいスープ、だしなどの食品についても、本来の味を損ねることなくコク味、濃厚感を強く付与できることが判明した。 From the results shown in Table 2, it was found that the present invention product can impart a strong taste and richness without sacrificing the original taste even for foods such as soups and soup stocks that are simple and easy to change in taste.
［官能試験５］乳製品への濃厚感付与効果
（１）アイスクリームへの添加効果
市販のアイスクリーム１００ｇに対し、酵母エキス２を０．１ｇ添加し、官能試験を行った。
[Sensory test 5] Effect of imparting richness to dairy products (1) Effect of addition to ice cream 0.1 g of yeast extract 2 was added to 100 g of commercially available ice cream, and a sensory test was performed.
（２）ミルクセーキへの添加効果
下記の処方にて常法によりミルクセーキを作った。ミルクセーキ１００ｍｌに対し、酵母エキス２を０．１ｇ添加し、官能試験を行った。
ミルクセーキの処方 ６５ｇ中
卵黄 ２０ｇ
牛乳 ３０ｇ
砂糖 １０ｇ
バニラエッセンス ２〜３滴
結果を表３に示す。
(2) Effect of addition to milkshake A milkshake was prepared by a conventional method with the following formulation. 0.1 g of yeast extract 2 was added to 100 ml of milkshake, and a sensory test was performed.
Milkshake prescription 65g egg yolk 20g
30g of milk
10g sugar
Table 2 shows the results of 2 to 3 drops of vanilla essence.
表３の結果から、本発明品は、乳製品のように旨味を必要としない食品に対し、味に変化を与えず、濃厚感を付与できることがわかった。 From the results of Table 3, it was found that the product of the present invention can give a rich feeling without changing the taste to foods that do not require umami like dairy products.
［官能試験６］辛味食品の濃厚感賦与効果
（１）キムチ鍋スープへの添加効果
市販のキムチ鍋スープ１００ｍｌに対し、酵母エキス２を０．５ｇ添加し、官能試験を行った。
[Sensory test 6] Concentration imparting effect of pungent food (1) Effect of adding to kimchi hot pot soup 100 g of commercial kimchi hot pot soup was added with 0.5 g of yeast extract 2 and subjected to a sensory test.
（２）カレーのルーへの添加効果
下記の処方にて常法によりカレーのルーを作った。
カレールー１００ｇに対し、酵母エキス２を０．５ｇ添加し、官能試験を行った。
カレールーの処方 １５０ｇ中
サラダ油 ６０ｇ
小麦粉 ７０ｇ
カレー粉 ２０ｇ
結果を表４に示す。
(2) Effect of adding curry roux to curry roux Curry roux was made by the conventional method with the following formulation.
0.5 g of yeast extract 2 was added to 100 g of Carreru, and a sensory test was performed.
60 g of salad oil in 150 g of Carreru prescription
70g flour
20g curry powder
The results are shown in Table 4.
表４の結果から、本発明品は、辛味製品に対しても味に変化を与えず、強い濃厚感を付与できることがわかった。 From the results of Table 4, it was found that the product of the present invention can give a strong rich feeling without changing the taste even for the pungent product.
［官能試験７］魚加工食品、畜肉加工食品の魚臭、畜肉臭マスキング効果
マスキング効果の官能試験は、比較対照として、酵母エキス無添加品及び酵母エキス７（低ペプチド品）を用い、１０名のパネラーにより官能試験を行い、一番強いマスキング効果を有すると判断したパネラーの数を官能試験の結果とした。
[Sensory test 7] Fish odor and processed meat odor masking effect of processed fish food and processed meat food The sensory test of masking effect uses yeast extract additive-free product and yeast extract 7 (low peptide product) as comparative controls. The sensory test was conducted with the panelists of No. 1, and the number of panelists judged to have the strongest masking effect was taken as the result of the sensory test.
（１）魚醤への添加効果
市販の魚醤１００ｍｌに対し、酵母エキス２または酵母エキス７を０．０１ｇ添加し、酵母エキス２添加、酵母エキス７添加、無添加の場合の３通りについて魚臭マスキング効果を評価した。
(1) Additive effect to fish soy 0.01 g of yeast extract 2 or yeast extract 7 is added to 100 ml of commercially available fish soy, and fish is added in three ways when yeast extract 2 is added, yeast extract 7 is added, and no additive is added. The odor masking effect was evaluated.
（２）ソーセージへの添加効果
下記の処方にて常法によりソーセージを作った。
ソーセージ１００ｇに対し、酵母エキス２または酵母エキス７を０．３ｇ添加し、酵母エキス２添加、酵母エキス７添加、無添加の場合の３通りについて畜肉臭マスキング効果を評価した。
ソーセージの処方 １５０ｇ中
豚ミンチ肉 ９１ｇ
大豆タンパク ５ｇ
澱粉 ４ｇ
食塩 １．６ｇ
ヒ゜ロリン酸ナトリウム ０．３ｇ
アスコルヒ゛ン酸 ０．１ｇ
タンパク加水分解物 ０．２ｇ
胡椒 ０．４ｇ
水 ４７．４ｇ
(2) Effect of addition to sausage Sausage was made by a conventional method with the following formulation.
0.3 g of yeast extract 2 or yeast extract 7 was added to 100 g of sausage, and the meat odor masking effect was evaluated for three cases of yeast extract 2 addition, yeast extract 7 addition, and no addition.
Sausage recipe 150g pork minced meat 91g
Soy protein 5g
4g starch
1.6g of salt
Sodium pyrophosphate 0.3g
Ascorbic acid 0.1g
Protein hydrolyzate 0.2g
Pepper 0.4g
47.4g of water
［官能試験８］大豆製品の大豆臭マスキング効果
大豆臭マスキング効果は、官能試験７と同様の官能試験を行い評価した。
（１）豆乳への添加効果
市販の無調製豆乳１００ｍｌに対し、酵母エキス２または酵母エキス７を０．０１ｇ添加し、酵母エキス２添加、酵母エキス７添加、無添加の場合の３通りについて大豆臭マスキング効果を評価した。
[Sensory Test 8] Soybean Odor Masking Effect of Soybean Products The soybean odor masking effect was evaluated by performing a sensory test similar to the sensory test 7.
(1) Additive effect to soy milk 0.01 g of yeast extract 2 or yeast extract 7 is added to 100 ml of commercially available unprepared soy milk, and soybeans are added in three ways when yeast extract 2 is added, yeast extract 7 is added, and no additive is added. The odor masking effect was evaluated.
（２）業務用大豆蛋白への添加効果
市販の業務用大豆蛋白（粉末品）を５重量％になるよう温水に溶かし、その溶液１００ｍｌに対し、酵母エキス２または酵母エキス７を０．１ｇ添加し、酵母エキス２添加、酵母エキス７添加、無添加の場合の３通りについて大豆臭マスキング効果を評価した。
官能試験７及び８の試験結果を表５に示す。
(2) Effect of addition to commercial soy protein Commercial commercial soy protein (powder product) was dissolved in warm water to 5% by weight, and 0.1 g of yeast extract 2 or yeast extract 7 was added to 100 ml of the solution. Then, the soybean odor masking effect was evaluated for three cases of yeast extract 2 addition, yeast extract 7 addition, and no addition.
The test results of sensory tests 7 and 8 are shown in Table 5.
表５の結果から、本発明品の酵母エキスは、飲食品の不快な臭いを低減することがわかった。また、ペプチド低含有酵母エキスは、不快臭が無添加と同程度であり、低減効果が全くないことがわかった。 From the results of Table 5, it was found that the yeast extract of the present invention reduces the unpleasant odor of food and drink. Moreover, it was found that the low peptide-containing yeast extract has the same level of unpleasant odor as that of no addition, and has no reduction effect.
［官能試験９］
官能試験１で用いたコンソメスープ１００ｍｌに対し、酵母エキス４を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 9]
To 100 ml of the consomme soup used in sensory test 1, 0.1 g of yeast extract 4 was added and kept at 60 ° C. to perform a sensory test.
［官能試験１０］
官能試験２で用いただし１００ｍｌに対し、酵母エキス５を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 10]
Used in sensory test 2, but 0.1 g of yeast extract 5 was added to 100 ml, and kept at 60 ° C. to perform a sensory test.
［官能試験１１］
官能試験１で用いたコンソメスープ１００ｍｌに対し、酵母エキス６を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 11]
To 100 ml of the consomme soup used in sensory test 1, 0.1 g of yeast extract 6 was added and kept at 60 ° C. to perform a sensory test.
［官能試験１２］
官能試験２のだし１００ｍｌに対し、酵母エキス７を０.１ｇ添加し、６０℃に保持し、官能試験を行った。
[Sensory test 12]
0.1 g of yeast extract 7 was added to 100 ml of the stock of sensory test 2 and kept at 60 ° C. to perform a sensory test.
［官能試験１３］
官能試験４で用いた麺つゆに、酵母エキス６を０．１ｇ添加し、官能試験を行った。
[Sensory test 13]
0.1 g of yeast extract 6 was added to the noodle soup used in sensory test 4 to perform a sensory test.
［官能試験１４］
官能試験４で用いた麺つゆに、市販の酵母エキスであるＳＫ酵母エキスＳ−３（日本製紙ケミカル（株）製）を０．１ｇ添加し、官能試験を行った。
[Sensory test 14]
0.1 g of SK yeast extract S-3 (manufactured by Nippon Paper Chemicals Co., Ltd.), which is a commercially available yeast extract, was added to the noodle soup used in sensory test 4 to perform a sensory test.
［官能試験１５］
官能試験４で用いた麺つゆに、市販の酵母エキスであるアロマイルド（興人（株）製）を０．１ｇ添加し、官能試験を行った。
[Sensory test 15]
To the noodle soup used in sensory test 4, 0.1 g of a commercially available yeast extract alomil (manufactured by Kojin Co., Ltd.) was added to perform a sensory test.
官能試験９〜官能試験１５までの官能試験の結果を表６に示す。 Table 6 shows the results of the sensory test from sensory test 9 to sensory test 15.
表６の結果から、ペプチド比率の低い酵母エキスは、食品へのコク味、濃厚感の付与効果がなく、呈味成分が多いと、旨味を増強し、食品の味を変えてしまうことがわかった。また、酵母エキス４〜７の旨味付加作用は、それらに含まれる遊離アミノ酸および呈味性５’−ヌクレオチド類の作用によるものと思われる。 From the results of Table 6, it can be seen that the yeast extract having a low peptide ratio has no effect of imparting a rich taste and richness to the food, and if there are many taste components, the umami is enhanced and the taste of the food is changed. It was. Moreover, the umami addition action of the yeast extracts 4 to 7 seems to be due to the action of free amino acids and taste-related 5'-nucleotides contained therein.
［官能試験１６］
（１）アイスクリームへの添加効果
官能試験５で用いたアイスクリーム１００ｇに対し、酵母エキス７を０．１ｇ添加し、官能試験を行った。
[Sensory test 16]
(1) Effect of addition to ice cream 0.1 g of yeast extract 7 was added to 100 g of ice cream used in sensory test 5 to perform a sensory test.
（２）ミルクセーキへの添加効果
官能試験５で用いたミルクセーキ１００ｍｌに対し、酵母エキス７を０．１ｇ添加し、官能試験を行った。
(2) Effect of addition to milkshake 0.1 g of yeast extract 7 was added to 100 ml of milkshake used in sensory test 5, and a sensory test was performed.
［官能試験１７］
（１）アイスクリームへの添加効果
官能試験５で用いたアイスクリーム１００ｇに対し、ＳＫ酵母エキスＳ−３（日本製紙ケミカル（株）製）を０．１ｇ添加し、官能試験を行った。
[Sensory test 17]
(1) Addition effect to ice cream 0.1 g of SK yeast extract S-3 (manufactured by Nippon Paper Chemicals Co., Ltd.) was added to 100 g of ice cream used in sensory test 5 to perform a sensory test.
（２）ミルクセーキへの添加効果
実施例５のミルクセーキ１００ｍｌに対し、ＳＫ酵母エキスＳ−３（日本製紙ケミカル（株）製）を０．１ｇ添加し、官能試験を行った。
(2) Effect of addition to milk shake 0.1 g of SK yeast extract S-3 (manufactured by Nippon Paper Chemical Co., Ltd.) was added to 100 ml of the milk shake of Example 5, and a sensory test was performed.
［官能試験１８］
（１）アイスクリームへの添加効果
官能試験５で用いたアイスクリーム１００ｇに対し、アロマイルド（興人（株）製）を０．１ｇ添加し、官能試験を行った。
[Sensory test 18]
(1) Effect of adding to ice cream 0.1 g of alomiled (manufactured by Kojin Co., Ltd.) was added to 100 g of ice cream used in sensory test 5 to perform a sensory test.
（２）ミルクセーキへの添加効果
官能試験５で用いたミルクセーキ１００ｍｌに対し、アロマイルド（興人（株）製）を０．１ｇ添加し、官能試験を行った。
官能試験１６〜官能試験１８の結果を表７に示す。
(2) Effect of addition to milkshake 0.1 g of aromoid (manufactured by Kojin Co., Ltd.) was added to 100 ml of the milkshake used in sensory test 5 to perform a sensory test.
The results of sensory test 16 to sensory test 18 are shown in Table 7.
表７の結果から、ペプチド比率の低い酵母エキスは、食品へのコク味、濃厚感の付与効果がなく、呈味成分が多いと、旨味を増強し、食品の味を変えてしまうことがわかった。また、酵母エキス７については、それに含まれる遊離アミノ酸および呈味性５’−ヌクレオチド類が旨味付与していると思われる。
乳製品などの旨味を必要としない食品については、旨味が付与されることは大きな欠点である。
From the results in Table 7, it can be seen that the yeast extract having a low peptide ratio has no effect of imparting a rich taste and richness to food, and if there are many taste components, it enhances umami and changes the taste of the food. It was. Moreover, about the yeast extract 7, it is thought that the free amino acid and taste 5'-nucleotide contained in it impart umami.
For foods that do not require umami such as dairy products, imparting umami is a major drawback.
［官能試験１９］
（１）キムチ鍋スープへの添加効果
官能試験６において用いたキムチ鍋スープ１００ｍｌに対し、酵母エキス７を０．５ｇ添加し、官能試験を行った。
[Sensory test 19]
(1) Effect of addition to kimchi hot pot soup 100 g of kimchi hot pot used in sensory test 6 was added with 0.5 g of yeast extract 7 and subjected to a sensory test.
（２）カレールーへの添加効果
官能試験６において用いたカレールー１００ｇに対し、酵母エキス７を０．５ｇ添加し、官能試験を行った。
(2) Effect of addition to Carrero The organoleptic test was performed by adding 0.5 g of yeast extract 7 to 100 g of Carrere used in Sensory test 6.
［官能試験２０］
（１）キムチ鍋スープへの添加効果
官能試験６において用いたキムチ鍋スープ１００ｍｌに対し、ＳＫ酵母エキスＳ−３（日本製紙ケミカル（株）製）を０．５ｇ添加し、官能試験を行った。
[Sensory test 20]
(1) Effect of addition to kimchi hot pot soup 100 g of kimchi hot pot used in sensory test 6 was added with 0.5 g of SK yeast extract S-3 (manufactured by Nippon Paper Chemical Co., Ltd.), and a sensory test was conducted. .
（２）カレールーへの添加効果
官能試験６において用いたカレールー１００ｇに対し、ＳＫ酵母エキスＳ−３（日本製紙ケミカル（株）製）を０．５ｇ添加し、官能試験を行った。
(2) Effect of addition to Carreru The organoleptic test was performed by adding 0.5 g of SK yeast extract S-3 (manufactured by Nippon Paper Chemicals Co., Ltd.) to 100 g of Carrere used in Sensory Test 6.
［官能試験２１］
（１）キムチ鍋スープへの添加効果
官能試験６において用いたキムチ鍋スープ１００ｍｌに対し、アロマイルド（興人（株）製）を０．５ｇ添加し、官能試験を行った。
[Sensory test 21]
(1) Effect of addition to kimchi hot pot soup 100 g of kimchi hot pot used in sensory test 6 was added with 0.5 g of aromoid (manufactured by Kojin Co., Ltd.), and a sensory test was performed.
（２）カレールーへの添加効果
官能試験６のカレールー１００ｇに対し、アロマイルド（興人（株）製）を０．５ｇ添加し、官能試験を行った。
官能試験１９〜官能試験２１の結果を表８に示す。
(2) Effect of addition to Carreru The organoleptic test was performed by adding 0.5 g of Alo-Mild (manufactured by Kojin Co., Ltd.) to 100 g of Carreru in Sensory Test 6.
The results of sensory test 19 to sensory test 21 are shown in Table 8.
表８の結果から、ペプチド比率の低い酵母エキスは、辛味食品への濃厚感の付与効果がなく、呈味成分が多いと、旨味を増強し、食品の味を変えてしまうことがわかった。また、酵母エキス７については、それに含まれる遊離アミノ酸および呈味性５’−ヌクレオチド類が旨味付与していると思われる。 From the results of Table 8, it was found that the yeast extract with a low peptide ratio has no effect of imparting a rich feeling to the pungent food, and if there are many taste components, the umami is enhanced and the taste of the food is changed. Moreover, about the yeast extract 7, it is thought that the free amino acid and taste 5'-nucleotide contained in it impart umami.
［酸性ペプチドの分画と各画分の官能試験］
（１）酸性ペプチドの分取方法
１．０重量％の酵母エキス３水溶液８〜１０ｇを陰イオン交換樹脂（Ａｍｂｅｒｌｉｔｅ ＩＲＡ４０２ＢＬ（酢酸型）オルガノ（株）社製）カラム（４００ｃｍ3）に通し、吸着物質と非吸着物質に分画した。樹脂に吸着した酸性物質（酸性タンパク質、酸性ペプチド、酸性アミノ酸、核酸関連物質）を、２Ｎの酢酸で溶出させて回収した。回収した溶液から酢酸をロータリーエバポレーターで除去した後、アミノ酸測定用ＨＰＬＣで測定し、酸性の遊離アミノ酸含量を測定した。また、酢酸溶出液中のタンパク質を全て加水分解し、アミノ酸測定用ＨＰＬＣで測定し、酸性の全アミノ酸含量を測定した。全アミノ酸含量から遊離アミノ酸含量を差し引くことにより、酸性のペプチド含量を算出した。
[Fractionation of acidic peptides and sensory test of each fraction]
(1) through acidic peptides preparative methods 1.0 wt% yeast extract 3 solution 8~10g anion exchange resin (Amberlite IRA402BL (manufactured acetate form) Organo Co.) column (400 cm 3), adsorption Fractionated into material and non-adsorbed material. Acidic substances (acidic proteins, acidic peptides, acidic amino acids, nucleic acid-related substances) adsorbed on the resin were recovered by elution with 2N acetic acid. After removing acetic acid from the collected solution with a rotary evaporator, it was measured with HPLC for amino acid measurement, and the acidic free amino acid content was measured. In addition, all proteins in the acetic acid eluate were hydrolyzed and measured by HPLC for amino acid measurement to measure the total content of acidic amino acids. The acidic peptide content was calculated by subtracting the free amino acid content from the total amino acid content.
（２）酸性ペプチドの分子量による分画
下記、表９に示す条件にて、ゲル濾過クロマトグラフィーを行い、分画した。酵母エキス３の酸性ペプチドの分子量分布を図１に示す。
(2) Fractionation based on molecular weight of acidic peptide Gel filtration chromatography was performed for fractionation under the conditions shown in Table 9 below. The molecular weight distribution of the acidic peptide of yeast extract 3 is shown in FIG.
分子量別に数万〜約５，０００（Ａ画分）、約５，０００〜約１，０００（Ｂ画分）、約１，０００〜約２００（Ｃ画分）、２００未満（Ｄ画分）の４フラクションに分画した。 By molecular weight, tens of thousands to about 5,000 (A fraction), about 5,000 to about 1,000 (B fraction), about 1,000 to about 200 (C fraction), less than 200 (D fraction) Were fractionated into 4 fractions.
表１０に、各フラクションの全体に占める割合（（重量／重量）％）を示す。 Table 10 shows the ratio ( (weight / weight) %) of each fraction.
（３）分子量画分別官能試験
上記４分画を、コンソメスープに添加した時の官能評価の結果を示す。
（３−１）コク味付与効果
添加量を酵母エキスの１／１０量（０．０１重量％）として、官能試験１と同様にして、コク味付与効果を評価した。結果は、表１１に示す。ペプチド分画無添加のものをブランクとした。
(3) Sensory test for molecular weight fractionation The results of sensory evaluation when the above four fractions are added to consomme soup are shown.
(3-1) Body taste imparting effect The body taste imparting effect was evaluated in the same manner as in the sensory test 1 with the addition amount of body richness being 1/10 amount (0.01 wt%) of the yeast extract. The results are shown in Table 11. A peptide-free fraction was used as a blank.
（３−２）マスキング効果
添加量を酵母エキスの１／１０量（０．０１重量％）として、官能試験７と同様にしてマスキング効果を評価した。結果は、表１２に示す。ペプチド分画無添加のものをブランクとした。
(3-2) Masking effect The masking effect was evaluated in the same manner as in the sensory test 7 with the addition amount of the masking effect being 1/10 amount (0.01% by weight) of the yeast extract. The results are shown in Table 12. A peptide-free fraction was used as a blank.
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| WO2013031571A1 (en) | 2011-08-26 | 2013-03-07 | 興人ライフサイエンス株式会社 | Yeast extract having taste-enhancing effect |
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| US11096408B2 (en) | 2011-08-26 | 2021-08-24 | Mitsubishi Corporation Life Sciences Limited | Yeast extract having taste-enhancing effect |
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