JP2019129795A - Flavor improver - Google Patents
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本発明は、特定のペプチドを含む酵母エキス、風味改良剤に関する。 The present invention relates to a yeast extract and a flavor improving agent containing a specific peptide.
近年の加工食品に用いられる調味料は、消費者の健康志向に高まりにより、自然な風味、食品添加物不使用などの食品が求められている。そのため、酵母エキス、ＨＶＰ（Hydrolyzed Vegetable Protein：植物性たん白加水分解物）、畜肉エキスなどの天然系調味料が用いられている。 Condiments used in processed foods in recent years have been demanded of foods with natural flavors and no use of food additives due to the growing health-consciousness of consumers. Therefore, natural seasonings such as yeast extract, HVP (Hydrolyzed Vegetable Protein: vegetable protein hydrolyzate), and livestock meat extract are used.
従来から知られている呈味性成分（グルタミン酸ナトリウム、５’−ヌクレオチド、糖類）のほか、ペプチド成分が、呈味性に重要な働きがあることが知られてきた。
例えば、酵母エキスは、食品素材、調味料用途に広く用いられている素材であり、アミノ酸、ペプチド、糖類、５’−ヌクレオチド等を含んでいる。そのため、グルタミン酸ナトリウム、イノシン酸、グアニル酸などの単体の呈味性物質と比較し、酵母エキスは、複雑な風味を付与することができる。
また、酵母エキスにも含まれるペプチドが、呈味性に寄与していることが知られている。例えば、グルタチオンに代表される、γ−グルタミルペプチドがコク味付与物質として知られている（特許文献１）。
酵母以外では、乳蛋白質を加水分解したペプチドに、酸味がマスキングと人工甘味料の後切れを改善する効果を有することが知られている（特許文献２）
In addition to the conventionally known taste components (sodium glutamate, 5′-nucleotide, saccharide), it has been known that peptide components have an important function on taste.
For example, yeast extract is a material widely used for food materials and seasonings, and includes amino acids, peptides, sugars, 5′-nucleotides and the like. Therefore, compared with simple taste substances such as sodium glutamate, inosinic acid, and guanylic acid, yeast extract can impart a complex flavor.
In addition, it is known that peptides contained in yeast extract contribute to taste. For example, γ-glutamyl peptide represented by glutathione is known as a body taste imparting substance (Patent Document 1).
In addition to yeast, it is known that the acidity of a peptide obtained by hydrolyzing milk protein has an effect of improving masking and after-cutting of artificial sweeteners (Patent Document 2).
しかし、酵母エキス中のペプチドにおいては、前述のようなペプチドにおいては、コク味付与に寄与していることは知られているが、どのような種類のペプチドに風味改善効果があるかは、不明な点が多い。従来は、ペプチドを多く含ませることにより、風味を改善させるような方法が多く、このような場合、風味に悪影響があるペプチドが含まれることもあるため、風味改善効果が弱まることがあった。さらに、ペプチドが修飾等されることにより、色のつくことがあり、使用できる食品に制限がある場合もあった。さらに、分子量の大きいペプチドが含まれる場合は、溶解性が悪化することもあった。 However, among the peptides in yeast extract, it is known that the above-mentioned peptides contribute to richness, but it is unclear what kind of peptides have a flavor improving effect. There are many points. Conventionally, there are many methods for improving the flavor by adding a large amount of peptide. In such a case, a peptide having a bad influence on the flavor may be included, and thus the flavor improving effect may be weakened. Furthermore, there are cases where the food is colored due to the modification of the peptide, and the food that can be used is limited. Furthermore, when a peptide having a large molecular weight is included, the solubility may be deteriorated.
以上のことから、本発明は、ペプチドを含み、前段の課題を解決した風味改良剤を得ることを課題とする。 From the above, an object of the present invention is to obtain a flavor improving agent that contains a peptide and solves the problems of the previous stage.
本発明者らは、上記課題を解決するために、種々検討し、特定のペプチドを含有させることにより、食品に添加した際に、風味改良効果が高いことを見出し、本発明を完成させた。 In order to solve the above-mentioned problems, the present inventors have made various studies and found that the effect of improving the flavor is high when added to foods by adding a specific peptide, thereby completing the present invention.
（１）水溶性荷電ペプチドを含有する酵母エキス、
（２）前記（１）の水溶性荷電ペプチドが、ペプチド中の酸性アミノ酸含量が２０％以上である酵母エキス、
（３）前記（２）の水溶性荷電ペプチドが、さらに、ペプチド中の塩基性アミノ酸含量が１４．５％以上である酵母エキス、
（４）以下の(a)~(c)を含む、水溶性荷電ペプチドを含有する請求項１〜３に記載の酵母エキスの製造方法、
(a) 食用酵母を担子菌産生酵素類による酵素処理に供する工程
(b) 工程(a)で生じた酵素処理液を固液分離して上清を回収する工程；
(c) 前記上清から荷電性ペプチド含有組成物を得る工程
（５）前記（１）の製造方法中、工程(a)で使用する担子菌産生酵素類は、ヒイロタケ（Ｐｙｃｎｏｐｏｒｕｓ ｃｏｃｃｉｎｅｕｓ）の培養物又は培養抽出物である請求項１の製造方法。
（６）前記（１）の製造方法中、b)の固液分離が、精密濾過によるものであり、精密濾過膜は０．０５〜０．２２μmの平均孔径を有することを特徴とする請求項１記載の製造方法。
(1) a yeast extract containing a water-soluble charged peptide,
(2) The water-soluble charged peptide according to (1), wherein the acidic amino acid content in the peptide is 20% or more,
(3) The water-soluble charged peptide according to (2), wherein the yeast extract further has a basic amino acid content of 14.5% or more in the peptide,
(4) The method for producing a yeast extract according to claims 1 to 3, comprising a water-soluble charged peptide, including the following (a) to (c):
(a) Process of subjecting edible yeast to enzyme treatment with basidiomycetous enzymes
(b) a step of solid-liquid separation of the enzyme-treated solution produced in step (a) and collecting the supernatant;
(c) Step of obtaining a charged peptide-containing composition from the supernatant (5) In the production method of (1), the basidiomycete-producing enzyme used in step (a) is a culture of Pynoporus coccineus Or the manufacturing method of Claim 1 which is a culture extract.
(6) In the production method of (1), the solid-liquid separation of b) is performed by microfiltration, and the microfiltration membrane has an average pore diameter of 0.05 to 0.22 μm. 1. The production method according to 1.
本発明を食品に添加した際、特定のペプチドを含むことにより、添加した食品の味の持続性が向上する。さらに、甘味と旨味を強め、濃厚感を付与する酵母エキスを提供することができる。さらに、溶解性の良いペプチドを多く含むため、清澄性のあるスープ等に添加した場合でも濁りの発生を抑制する酵母エキスとなる。
When the present invention is added to a food, the taste sustainability of the added food is improved by including a specific peptide. Furthermore, the yeast extract which strengthens sweetness and umami | taste and provides a rich feeling can be provided. Furthermore, since it contains a lot of peptides with good solubility, it becomes a yeast extract that suppresses the occurrence of turbidity even when added to a clear soup or the like.
本発明は、水溶性荷電ペプチドを含有する風味改良剤であり、タンパク質を含む食品に担子菌産生酵素類による酵素処理に供する工程により、水溶性荷電ペプチドを含有する風味改良剤を得ることができる。タンパク質を含む食品としては、酵母を利用することが好ましい。 The present invention is a flavor improving agent containing a water-soluble charged peptide, and a flavor improving agent containing a water-soluble charged peptide can be obtained by subjecting a food containing protein to an enzyme treatment with basidiomycetes-producing enzymes. . As food containing protein, it is preferable to use yeast.
本発明に用いる酵母としては、食用酵母であれば特に限定するものではなく、生酵母、自体公知の方法で適宜乾燥した乾燥酵母いずれでもよく、例えば、ワイン酵母、パン酵母、清酒酵母、ビール酵母（Saccharomyces pastorianus）、トルラ酵母（Candida utilis）等が使用できる。特に、トルラ酵母が好ましい。 The yeast used in the present invention is not particularly limited as long as it is an edible yeast, and may be live yeast or dry yeast appropriately dried by a method known per se, such as wine yeast, baker's yeast, sake yeast, and brewer's yeast. (Saccharomyces pastorianus), Torula yeast (Candida utilis), etc. can be used. In particular, Torula yeast is preferred.
酵母の培養方法は、制限なく、通常の方法で培養したものを使用することができる。一般的には、酵母を培養する際の培地には、炭素源として、ブドウ糖、酢酸、エタノール、グリセロール、糖蜜、亜硫酸パルプ廃液等が用いられ、窒素源としては、尿素、アンモニア、硫酸アンモニウム、塩化アンモニウム、硝酸塩などが使用される。リン酸、カリウム、マグネシウム源も過リン酸石灰、リン酸アンモニウム、塩化カリウム、水酸化カリウム、硫酸マグネシウム、塩化マグネシウム等の通常の工業用原料でよく、その他亜鉛、銅、マンガン、鉄イオン等の無機塩を添加する。その他は、ビタミン、アミノ酸、核酸関連物質等を使用しないでも培養可能であるが、これらを添加しても良い。コーンスチーブリカー、カゼイン、酵母エキス、肉エキス、ペプトン等の有機物を添加しても良い。また、市販されている乾燥酵母を使用してもよい。 The culture method of yeast can use what was culture | cultivated by the normal method without a restriction | limiting. Generally, glucose, acetic acid, ethanol, glycerol, molasses, sulfite pulp waste liquid, etc. are used as the carbon source in the medium for culturing yeast, and urea, ammonia, ammonium sulfate, ammonium chloride are used as the nitrogen source. Nitrates and the like are used. Phosphoric acid, potassium, and magnesium sources may be ordinary industrial raw materials such as lime perphosphate, ammonium phosphate, potassium chloride, potassium hydroxide, magnesium sulfate, magnesium chloride, and other zinc, copper, manganese, iron ions, etc. Add inorganic salt. Others can be cultured without using vitamins, amino acids, nucleic acid-related substances, etc., but these may be added. Organic substances such as corn steep liquor, casein, yeast extract, meat extract and peptone may be added. Moreover, you may use the dry yeast marketed.
培養温度やｐＨ等の培養条件は、特に制限なく適用でき、使用する酵母菌株に合わせて設定すれば良い。一般的には、培養温度は２１〜３７℃、好ましくは２５〜３４℃が良く、ｐＨは３．０〜８．０、特に３．５〜７．０が好ましい。 Culture conditions such as culture temperature and pH can be applied without particular limitation, and may be set according to the yeast strain to be used. Generally, the culture temperature is 21 to 37 ° C, preferably 25 to 34 ° C, and the pH is 3.0 to 8.0, particularly 3.5 to 7.0.
本発明の培養形式としては、バッチ培養、あるいは連続培養のいずれでも良いが、工業的には後者が望ましい。培養時の撹拌、通気等の条件は特に制限なく、一般的な方法でよい。 The culture format of the present invention may be either batch culture or continuous culture, but the latter is desirable industrially. Conditions such as agitation and aeration during culture are not particularly limited and may be a general method.
菌体培養後に本発明のエキス抽出を行う。菌体培養後の湿潤酵母菌体を蒸留水に懸濁して遠心分離を繰り返すことで洗浄した後に、酵母エキスの抽出を行う。
本発明の抽出法は、ｐＨ７．０〜８．５に調整し、５０〜９０℃で１０〜６０分間加熱撹拌して洗浄する。ｐＨ調整は、通常の方法でよい。洗浄後、再度遠心分離をする。遠心分離で得られた沈殿物を再懸濁し、酵素処理をする。
再懸濁は、菌体濃度が乾燥重量換算で、５〜３０％、好ましくは１０〜２０％になるように蒸留水に再懸濁する。
The extract of the present invention is extracted after culturing the cells. The wet yeast cells after culturing the cells are suspended in distilled water and washed by repeated centrifugation, and then the yeast extract is extracted.
In the extraction method of the present invention, the pH is adjusted to 7.0 to 8.5, and the mixture is washed by heating and stirring at 50 to 90 ° C. for 10 to 60 minutes. The pH may be adjusted by a usual method. After washing, centrifuge again. The precipitate obtained by centrifugation is resuspended and treated with an enzyme.
The resuspension is resuspended in distilled water so that the bacterial cell concentration is 5 to 30%, preferably 10 to 20% in terms of dry weight.
次いで、得られた懸濁液に酵素処理をする。本発明で使用する酵素は、プロテアーゼである。本発明で使用するプロテアーゼには、担子菌産生酵素類を使用する。例えば、ホウロクタケ属に属する担子菌、好ましくはヒイロタケ（Ｐｙｃｎｏｐｏｒｕｓ ｃｏｃｃｉｎｅｕｓ）を公知の方法により培養し、プロテアーゼを含有する培養液、培養抽出物、濃縮物、又は乾燥物等を使用する。さらには、公知の方法により、プロテアーゼ必須構成酵素とする酵素類を採取し、粗製酵素、または精製酵素として用いることができる。また、担子菌産生酵素は、プロテアーゼ活性だけでなく、グルカナーゼ活性を有しているため、酵素に適用した際には、細胞壁成分も分解可能である。 Next, the obtained suspension is subjected to enzyme treatment. The enzyme used in the present invention is a protease. As the protease used in the present invention, basidiomycete-producing enzymes are used. For example, basidiomycetes belonging to the genus Pleurotus, preferably Pycnoporus coccineus, are cultured by a known method, and a culture solution, a culture extract, a concentrate, or a dried product containing protease are used. Furthermore, enzymes that are protease-constituting enzymes can be collected by known methods and used as crude enzymes or purified enzymes. In addition, the basidiomycete-producing enzyme has not only protease activity but also glucanase activity, so that when applied to the enzyme, cell wall components can be decomposed.
担子菌産生酵素類の添加量は、酵素の調整方法により、通常の酵素と同様に適宜調整し添加する。水溶液の水溶性部分の乾燥物を用いる場合、酵母に対して、０．３〜１．５重量％程度使用する。 The amount of basidiomycete-producing enzymes is appropriately adjusted and added in the same manner as normal enzymes, depending on the enzyme adjustment method. When using the dried material of the water-soluble part of aqueous solution, it uses about 0.3 to 1.5 weight% with respect to yeast.
本発明では、必要に応じ、５’−イノシン酸及び５’−グアニル酸などの呈味性５’−ヌクレオチド類を含有させる工程を含める。通常は、ヌクレアーゼ、デアミナーゼ処理をすることで、酵母中の核酸を分解することで、本発明の風味改良剤に５’−イノシン酸及び５’−グアニル酸を含有さることができる。酵素は、通常の酵母エキス製造に用いるものでよい。本発明では、ヌクレアーゼ、デアミナーゼ反応後、前記の担子菌産生酵素類を添加し、処理する。また、酵母の自己消化法と組み合わせても良い。添加する酵素量、酵素反応の温度、ｐＨ、反応時間は各酵素により異なるが、各酵素の至適温度、至適ｐＨで反応させることが好ましい。自己消化法は、一般的な方法でよい。 In the present invention, a step of containing tasty 5'-nucleotides such as 5'-inosinic acid and 5'-guanylic acid is included as necessary. Usually, the flavor improving agent of the present invention can contain 5'-inosinic acid and 5'-guanylic acid by degrading nucleic acids in yeast by nuclease and deaminase treatment. An enzyme may be used for normal yeast extract manufacture. In the present invention, after the nuclease and deaminase reaction, the basidiomycete-producing enzymes are added and processed. Moreover, you may combine with the self-digestion method of yeast. The amount of enzyme to be added, the temperature, pH of the enzyme reaction, and the reaction time vary depending on each enzyme, but it is preferable to react at the optimum temperature and pH of each enzyme. The self-digestion method may be a general method.
本発明では、上記酵素と酵母との反応は、同一反応容器内で連続的に、反応条件を変えることも可能である。例えば、酵母に対し、ヌクレアーゼ、デアミナーゼを、ｐＨ６．５〜８．０にて、０．１〜３．０重量％添加し、３５℃〜６０℃ で３〜１０時間反応させ、次に６０℃〜７０℃で１〜６時間酵母と接触させる。その後、ｐＨ３．５〜５．５にて、担子菌産生酵素類を０．３〜１．５重量％添加し、４５℃〜６０℃で５〜２０時間酵母と接触させる。ｐＨ調整は、通常の方法でよく、必要に応じて酸（塩酸等） またはアルカリ（水酸化ナトリウム等） を用いて行なう。酵素反応後、必要に応じて、酵素の失活処理を行う。通常は、９０〜１００℃で１０〜６０分間加熱することにより、酵素を失活させる。 In the present invention, the reaction between the enzyme and yeast can be continuously changed in the same reaction vessel. For example, nuclease and deaminase are added to yeast at pH 6.5 to 8.0 and 0.1 to 3.0% by weight, and reacted at 35 ° C to 60 ° C for 3 to 10 hours. Contact with yeast at ˜70 ° C. for 1-6 hours. Thereafter, at pH 3.5 to 5.5, basidiomycete-producing enzymes are added in an amount of 0.3 to 1.5% by weight and brought into contact with yeast at 45 ° C to 60 ° C for 5 to 20 hours. The pH may be adjusted by a usual method, and an acid (such as hydrochloric acid) or an alkali (such as sodium hydroxide) is used as necessary. After the enzyme reaction, the enzyme is deactivated as necessary. Usually, the enzyme is inactivated by heating at 90 to 100 ° C. for 10 to 60 minutes.
酵素反応後、遠心分離、濾過等により、固体分を除去する。遠心分離は、常法でよい。濾過の場合は、精密濾過を行うとよい。精密濾過も、公知の方法でよい。使用する平均孔０．０５〜０．２μｍの精密濾過膜で行うのが好ましい。 After the enzyme reaction, the solid content is removed by centrifugation, filtration or the like. Centrifugation may be performed by a conventional method. In the case of filtration, microfiltration may be performed. Microfiltration may also be a known method. It is preferable to use a microfiltration membrane having an average pore size of 0.05 to 0.2 μm.
固形分除去後、そのまま風味改良剤として使用することができるが、濃縮、乾燥により、粉末化しても良い。ろ液の固形分濃度を１０〜５０重量％、好ましくは３０〜４０重量％ に濃縮してもよい。濃縮方法は特に限定するものではなく、例えば、常圧加熱濃縮、減圧過熱濃縮、冷凍濃縮等の公知の濃縮方法が採用できる。乾燥方法も公知の方法により、乾燥、粉末化してもよい。特に、５０℃ 以上に加温して噴霧乾燥することが、濃縮液のゲル化防止法として好ましい。 After removing the solid content, it can be used as it is as a flavor improving agent, but it may be powdered by concentration and drying. You may concentrate the solid content concentration of a filtrate to 10 to 50 weight%, Preferably it is 30 to 40 weight%. The concentration method is not particularly limited, and for example, a known concentration method such as atmospheric pressure heating concentration, reduced pressure overheating concentration, or freezing concentration can be employed. The drying method may be dried and powdered by a known method. In particular, heating to 50 ° C. or higher and spray drying is preferable as a method for preventing gelation of the concentrated solution.
本発明で、水溶性荷電ペプチドとは、ペプチド中の酸性アミノ酸含量が２０％以上及び／又は水溶性ペプチド中の塩基性アミノ酸含量が１４．５％以上となるペプチドである。酸性アミノ酸は、アスパラギン酸とグルタミン酸である。塩基性アミノ酸は、リシン、ヒスチジン、アルギニンである。本発明でペプチド含量は、本発明でいうペプチドとは、全アミノ酸量より遊離アミノ酸量を引くことにより算出する。本発明でのペプチド含量は、固形分あたり、３０重量％以上含有する。特に好ましくは４０重量％以上である。 In the present invention, the water-soluble charged peptide is a peptide having an acidic amino acid content of 20% or more in the peptide and / or a basic amino acid content of 14.5% or more in the water-soluble peptide. Acidic amino acids are aspartic acid and glutamic acid. Basic amino acids are lysine, histidine, arginine. In the present invention, the peptide content is calculated by subtracting the free amino acid amount from the total amino acid amount with the peptide referred to in the present invention. The peptide content in the present invention is 30% by weight or more per solid content. Especially preferably, it is 40 weight% or more.
試料中のペプチド組成は、全アミノ酸と遊離アミノ酸をアミノ酸分析計（日立高速アミノ酸分析計Ｌ−８８００）で測定し、全アミノ酸−遊離アミノ酸をペプチド組成とした。 The peptide composition in the sample was determined by measuring all amino acids and free amino acids with an amino acid analyzer (Hitachi High-Speed Amino Acid Analyzer L-8800), and using all amino acids-free amino acids as the peptide composition.
各アミノ酸の含有量は、アミノ酸測定用ＨＰＬＣにより、含有アミノ酸分析を行う。ペプチド中に含まれる各アミノ酸の含有量を測定後、各アミノ酸の構成比から、酸性アミノ酸のアスパラギン酸とグルタミン酸、塩基性アミノ酸のリシン、ヒスチジン及びアルギニンの構成比を加算し、アスパラギン酸とグルタミン酸の含有量がペプチドを構成する全アミノ酸中２０％以上、リシン、ヒスチジン及びアルギニンが１５％以上となるのが、本願発明の風味改良剤である。本発明の機能を発揮するには、ペプチド中の塩基性アミノ酸含量は高い方がよい。ペプチド中の酸性アミノ酸の含量は、２０％以上であるが、さらに好ましくは、２０％以上３５％未満である。特に好ましくは２５％以上３５％未満である。 The content of each amino acid is analyzed by amino acid measurement HPLC. After measuring the content of each amino acid contained in the peptide, from the composition ratio of each amino acid, add the composition ratios of the acidic amino acids aspartic acid and glutamic acid, the basic amino acids lysine, histidine and arginine, and the aspartic acid and glutamic acid In the flavor improving agent of the present invention, the content is 20% or more of all amino acids constituting the peptide, and lysine, histidine and arginine are 15% or more. In order to exhibit the function of the present invention, the basic amino acid content in the peptide should be high. The content of acidic amino acids in the peptide is 20% or more, more preferably 20% or more and less than 35%. Particularly preferably, it is 25% or more and less than 35%.
本発明では、前段までのペプチドのほかに、呈味性５’−ヌクレオチド類を含有させる。含有量としては、５’−イノシン酸及び５’−グアニル酸の合計量で、３重量％以上含有することが好ましい。さらに好ましくは、本発明の特定のペプチドとのバランスから３〜８重量％含有させる。呈味性５’−ヌクレオチド類と本発明のペプチドを含有することにより、甘味と旨味の持続性が発揮される。 In the present invention, in addition to the peptides up to the previous stage, a tasty 5'-nucleotide is included. The total content of 5'-inosinic acid and 5'-guanylic acid is preferably 3% by weight or more. More preferably, it is contained in an amount of 3 to 8% by weight from the balance with the specific peptide of the present invention. By containing the tasty 5'-nucleotides and the peptide of the present invention, sweetness and umami persistence are exhibited.
本発明の製造法で得られた風味改良剤は、公知の風味改良剤と同様に使用することができ、例えば、得られた風味改良剤を農産加工食品（野菜、果実、穀物等の加工品を含む）、水産加工食品（魚介類、海藻等の加工品を含む）、畜産加工食品（畜肉・卵・乳製品等の加工品を含む）、だし・つゆ・ソース・醤油・みそ、あらゆる合わせ調味料等に使用することができる。 The flavor improver obtained by the production method of the present invention can be used in the same manner as known flavor improvers. For example, the obtained flavor improver is processed into agricultural processed foods (processed products such as vegetables, fruits, and grains). Seafood processed foods (including processed products such as seafood and seaweed), livestock processed foods (including processed products such as livestock meat, eggs and dairy products), dashi, soy sauce, sauce, soy sauce and miso It can be used as a seasoning.
本発明の風味改良剤は、上記食品に対して、０．０１〜０．５重量％となるよう添加する。本発明の風味改良剤は、添加した食品の甘味と旨味の持続性を向上させる。当該効果は、水溶性荷電ペプチドによる効果であると考えている。 The flavor improving agent of this invention is added so that it may become 0.01 to 0.5 weight% with respect to the said foodstuff. The flavor improving agent of the present invention improves the sustainability of the sweetness and umami of the added food. This effect is considered to be due to the water-soluble charged peptide.
（実施例１）
（ヒイロタケ産生酵素類の調製）
（ １ ） ヒイロタケ（Ｐｙｃｎｏｐｏｒｕｓ ｃｏｃｃｉｎｅｕｓ） の胞子懸濁液（１０ ７個／ｍｌ 以上）２ｍｌを種培地（塩化カルシウム１ｇ／ｌ、硫酸マグネシウム１ｇ／ｌ、硫酸アンモニウム２ｇ／ｍｌ、リン酸一カリウム２ｇ ／ｌ、ショ糖５０ｇ／ｌ、コーン・スティープ・リカー３０ｇ／ｌ、ｐ Ｈ７ ．０）２０ｍｌに接種し、２００ｍｌ容フラスコ中２８℃ 、２００ｒｐｍで４８ 時間培養し、種培養終了液を得た。
（ ２ ） 得られた種培養終了液２ ｍｌを主培地（塩化カルシウム１ｇ ／ｌ 、硫酸マグネシウム１ｇ ／ｌ、硫酸アンモニウム２ｇ／ｌ、リン酸一カリウム２ｇ／ｌ、ショ糖８０ ｇ／ｌ、脱脂大豆粉３５ｇ／ｌ 、ｐＨ６．０）２０ｍｌに移植し、２００ｍｌ容フラスコで２８℃、２００ｒｐｍで９６時間培養し、主培養終了液を得た。主培養終了液を、濾紙で濾過し、得られた酵素液を真空乾燥してヒイロタケ産生酵素類の乾燥粉末を得た。
Example 1
(Preparation of oyster mushroom producing enzymes)
(1) 2 ml of spore suspension of Pycnoporus coccineus (10 7 pieces / ml or more) in a seed medium (calcium chloride 1 g / l, magnesium sulfate 1 g / l, ammonium sulfate 2 g / ml, monopotassium phosphate 2 g / l Sucrose 50 g / l, corn steep liquor 30 g / l, pH 7.0) and inoculated 20 ml, and cultured in a 200 ml flask at 28 ° C. and 200 rpm for 48 hours to obtain a seed culture end solution.
(2) 2 ml of the seed culture end solution obtained was added to the main medium (calcium chloride 1 g / l, magnesium sulfate 1 g / l, ammonium sulfate 2 g / l, monopotassium phosphate 2 g / l, sucrose 80 g / l, defatted soybeans) This was transplanted into 20 ml of powder 35 g / l, pH 6.0), and cultured in a 200 ml flask at 28 ° C. and 200 rpm for 96 hours to obtain a main culture end liquid. The main culture completion liquid was filtered with a filter paper, and the obtained enzyme liquid was vacuum-dried to obtain a dried powder of agaric-producing enzymes.
市販の乾燥キャンディダ・ユティリスを使用し、当該菌体で１０％菌体懸濁液１０００ｍｌを常法でｐＨ７．５に調整し、６０℃、３０分間加熱処理した後、遠心分離で菌体を回収し、菌体を水で洗浄し余分な抽出物を除去した。本菌体を純水で菌体濃度１０％に調整した。これを常法により、ｐＨ７．７に調整した。調整後、酵母固形分に対して０ ．２５％の割合で、リボヌクレアーゼＰ （天野エンザイム製５’−フォスフォジエステラーゼ）の３０ｍｇ を少量の水に溶解して加え、同温度で撹拌しながら３時間反応させた。次いで、液温を４５℃として、デアミザイム（天野エンザイム製デアミナーゼ）の２０ｍｇを少量の水に溶解して加え、この温度下で２時間撹拌しながら保持した。４０℃ から６５℃ まで５時間で昇温させ、ついで６５℃で３時間保持した。反応後、５０℃に冷却し、ｐＨ４．０に調整し、酵母固形分に対して０．６ ％ の割合で、上記で得られた担子菌産生酵素乾燥粉末を加え、５０℃ で１２時間反応させた。
反応終了後、９０℃で１０分間加熱殺菌し、６０℃に冷却した。反応液を０．１〜０．２μｍの孔径を有する精密濾過膜で濾過した。残渣を３倍量の純水と共に撹拌し、再度濾過して、濾液を上記の濾液と合した。濾過後、濃縮、乾燥さえ、風味改良剤を得た。ペプチド含量は、４２重量％、５’−イノシン酸及び５’−グアニル酸の合計含量は、６．６重量％であった。
得られた風味改良剤中でペプチドを構成するアミノ酸分析値を表１に示す。
Using commercially available dried Candida utilis, 1000 ml of a 10% cell suspension with the cells was adjusted to pH 7.5 by a conventional method, heat-treated at 60 ° C. for 30 minutes, and then centrifuged to remove the cells. The cells were collected and the cells were washed with water to remove excess extract. This bacterial cell was adjusted to a bacterial cell concentration of 10% with pure water. This was adjusted to pH 7.7 by a conventional method. After adjustment, 0. At a rate of 25%, 30 mg of ribonuclease P (Amanoenzyme 5′-phosphodiesterase) was dissolved in a small amount of water, and the mixture was reacted at the same temperature for 3 hours with stirring. Next, the liquid temperature was adjusted to 45 ° C., 20 mg of deamizyme (deaminase manufactured by Amano Enzyme) was dissolved in a small amount of water, and the mixture was held at this temperature with stirring for 2 hours. The temperature was raised from 40 ° C. to 65 ° C. over 5 hours, and then held at 65 ° C. for 3 hours. After the reaction, it is cooled to 50 ° C., adjusted to pH 4.0, the basidiomycete-producing enzyme dry powder obtained above is added at a ratio of 0.6% to the yeast solid content, and the reaction is carried out at 50 ° C. for 12 hours. I let you.
After completion of the reaction, the mixture was sterilized by heating at 90 ° C. for 10 minutes and cooled to 60 ° C. The reaction solution was filtered through a microfiltration membrane having a pore size of 0.1 to 0.2 μm. The residue was stirred with 3 volumes of pure water, filtered again, and the filtrate was combined with the above filtrate. After filtration, a flavor improver was obtained even after concentration and drying. The peptide content was 42% by weight, and the total content of 5′-inosinic acid and 5′-guanylic acid was 6.6% by weight.
Table 1 shows the analysis values of amino acids constituting peptides in the obtained flavor improving agent.
（実施例２）
実施例１と同様に酵母菌体を調整し、リボヌクレアーゼ、デアミザイム処理をした。反応後、５５℃に冷却し、ｐＨ４．５に調整し、酵母固形分に対して１．０％の割合で、上記で得られた担子菌産生酵素乾燥粉末を加え、５５℃ で１２時間反応させた。反応後は、実施例１と同様に、処理し、アミノ酸分析をした、アミノ酸分析値は、表１に示す。ペプチド含量は、４１重量％、５’−イノシン酸及び５’−グアニル酸の合計含量は、５．８重量％であった。
(Example 2)
Yeast cells were prepared in the same manner as in Example 1 and treated with ribonuclease and deamizyme. After the reaction, the reaction mixture is cooled to 55 ° C., adjusted to pH 4.5, and the dried basidiomycete-producing enzyme dry powder obtained above is added at a rate of 1.0% with respect to the yeast solid content and reacted at 55 ° C. for 12 hours. I let you. After the reaction, treatment and amino acid analysis were performed in the same manner as in Example 1. The amino acid analysis values are shown in Table 1. The peptide content was 41% by weight, and the total content of 5′-inosinic acid and 5′-guanylic acid was 5.8% by weight.
比較として、市販の乾燥ビール酵母も実施例１と同様に酵素反応させ、酵母エキスを製造した（比較例１）。当該酵母エキスのペプチドを構成するアミノ酸分析値を表１に示す。ペプチド含量は、２７重量％、５’−イノシン酸及び５’−グアニル酸の合計含量は、３．２重量％であった。
実施例１及び２で得られた、風味改良剤と比較例１の酵母エキスの添加効果を確認した。
（ホワイトソース）
表２に示す処方で対照のホワイトソースを作成した。先ず、鍋にバターと小麦粉を入れ、中火で良く炒めた。ここに残りの材料を入れてとろみが付くまで煮込んだ。このホワイトソース１００．０ｇに、実施例１、２及び比較例１の改良剤を０．１ｇ入れて溶解し、これを評価用サンプルとした。
The effects of adding the flavor improver obtained in Examples 1 and 2 and the yeast extract of Comparative Example 1 were confirmed.
(White sauce)
A control white sauce was prepared according to the formulation shown in Table 2. First, put butter and flour in a pan and fry well over medium heat. The remaining ingredients were put here and simmered until thick. In 100.0 g of this white sauce, 0.1 g of the improving agent of Examples 1 and 2 and Comparative Example 1 was added and dissolved, and this was used as a sample for evaluation.
官能評価により、下記表の配合で、ホワイトソースの食味を評価した。10名の訓練されたパネラーにより、実施例で得られた風味改良剤と酵母エキスを添加し、無添加のものと比較し、添加効果の評価を行った。その結果、実施例１及び２では、甘味、旨味が強くなり、その持続性があった。比較例１では、先味である旨味を強く感じるが、対照サンプルと比較し、甘味には変化がなかった。 The taste of the white sauce was evaluated by sensory evaluation with the composition shown in the following table. The taste improver and yeast extract obtained in the Examples were added by 10 trained panelists, and the effect of addition was evaluated in comparison with the additive-free one. As a result, in Examples 1 and 2, sweetness and umami became strong and sustained. In Comparative Example 1, the taste of taste was strongly felt, but there was no change in sweetness compared to the control sample.
Claims (6)
(d) 食用酵母を担子菌産生酵素類による酵素処理に供する工程
(e) 工程(a)で生じた酵素処理液を固液分離して上清を回収する工程；
(f) 前記上清から荷電性ペプチド含有組成物を得る工程
The method for producing a yeast extract according to any one of claims 1 to 3, comprising a water-soluble charged peptide comprising the following (a) to (c):
(d) Process of subjecting edible yeast to enzyme treatment with basidiomycetes-producing enzymes
(e) a step of solid-liquid separation of the enzyme-treated solution produced in step (a) and collecting the supernatant;
(f) obtaining a charged peptide-containing composition from the supernatant
The method according to claim 1, wherein the basidiomycete-producing enzymes used in step (a) in the production method of claim 1 are a culture or extract of Pynoporus coccineus.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022203049A1 (en) * | 2021-03-26 | 2022-09-29 | 三菱商事ライフサイエンス株式会社 | Yeast extract achieving spread of flavor and production method therefor |
| WO2023281779A1 (en) * | 2021-07-05 | 2023-01-12 | 三菱商事ライフサイエンス株式会社 | Off-flavor suppressing yeast extract |
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| JPH0848698A (en) * | 1994-08-05 | 1996-02-20 | Seiwa Kasei:Kk | Peptide composition derivatized from yeast protein, its production and its use |
| JP2004229540A (en) * | 2003-01-29 | 2004-08-19 | Takeda-Kirin Foods Corp | Method for producing yeast extract |
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| JPS62275669A (en) * | 1986-05-26 | 1987-11-30 | Sapporo Breweries Ltd | Production of yeast extract |
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| WO2023281779A1 (en) * | 2021-07-05 | 2023-01-12 | 三菱商事ライフサイエンス株式会社 | Off-flavor suppressing yeast extract |
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