JP5215518B2

JP5215518B2 - Enzymatic degradation product and production method thereof

Info

Publication number: JP5215518B2
Application number: JP2002501275A
Authority: JP
Inventors: 定治横山; 彰二垂水
Original assignee: Takara Shuzo Co Ltd
Current assignee: Takara Shuzo Co Ltd
Priority date: 2000-06-02
Filing date: 2001-05-31
Publication date: 2013-06-19
Anticipated expiration: 2021-05-31
Also published as: WO2001093699A1; JPWO2001093699A1

Description

技術分野
本発明は、調味料原料に、特定の条件下で蛋白質分解酵素を作用させてなる酵素分解物、該酵素分解物からなる調味料及びそれらの製造方法に関する。

背景技術
従来、原料を塩酸等で加水分解することで種々の調味料が製造されているが、ＭＣＰ、ＤＣＰ等の有害な塩素化合物も副生されることから、近年の食品の安全性への関心の高まりと共に、酵素分解によって調味料を製造することが注目されている。例えば、魚を６０℃以上の温度に加熱して魚体中に含まれる自己消化酵素を不活性化し、次いで５０〜６０℃、ｐＨ９〜１０において耐アルカリ性蛋白質分解酵素を添加して分解した後、ｐＨを５〜６に調整し別の耐酸性蛋白質分解酵素を添加して同じく５０〜６０℃で分解する調味料の製造方法（特公昭５３−３１９３５号公報）、削り節の熱水抽出残渣を、エンドタイプのプロテアーゼを中性で作用させて蛋白質を可溶化させた後、エキソタイプのプロテアーゼを酸性にて作用させて苦味成分を分解し、旨み成分を増加させることを特徴とする液体調味料の製造方法（特開平５−３０８９２２号公報）、小麦グルテン、動物蛋白（ゼラチン）等を５０〜６０℃条件下で酵素分解を行い調味料を製造する方法（食品と開発、３１、２、２０〜２２頁、１９９６年）がある。これら従来の酵素分解においては、酵素分解反応液中の雑菌汚染を防止するために、該酵素分解反応液中の温度を５０〜６０℃という高温に設定しなければならず、この結果、得られる製品に加熱臭が付くことが避けられないという問題点が存在した。また、一般に使用されるエキソ型プロテアーゼは糸状菌由来であり、耐熱性が低く、該酵素分解反応液中の温度が５５℃を越えると酵素活性が次第に失活してしまう。そのため実生産においては、温度管理、かくはん能力等を備えた酵素分解反応槽が必要であり、かなりの設備投資、生産管理要員及びメンテナンス費用を要する点が問題となっている。
一方、魚醤油の製造方法において、蛋白質分解酵素及びエタノール濃度０．５〜５％（ｖ／ｖ）に相当するエタノールの存在下で、無機塩濃度３〜１２％（ｗ／ｗ）として原料魚介を酵素分解する方法が開示されている（特開平３−２５４６６０号公報）。しかし、この方法は、一般的な食品用酵素を魚醤油の製造のための一般的な方法で（すなわち、エタノール、無機塩などの存在下で）使用している。この方法における条件は、酵素の特性（至適ｐＨを含む）を生かした効果的な酵素の使用には適していないので、原料利用率が低いという欠点を有する。特に、だし汁製造企業等から排出される抽出残渣の有効利用技術の開発が求められている現状から、原料利用率の高い産業廃棄物利用技術の開発が求められている。
また、コーングルテンミール（ＣＧＭ）はコーンスターチの副産物として大量に（日本国内で年間約２０万トン）産出され、蛋白質を多量に含有する資源である。しかし、ＣＧＭは分解されにくいので、現在では、家畜の飼料などに利用されているのみである。従って、ＣＧＭのより有効な利用方法の開発が望まれている。

発明の目的
本発明の第１の目的は、特別な設備を要しない簡易な設備を用いて、調味料原料を酵素分解した酵素分解物を提供することにあり、第２の目的は、第１の目的の酵素分解物の製造方法を提供することにある。

発明の概要
本発明を概説すると、調味料原料に、いずれも、酵母及び乳酸菌が生育しない条件下において、（ａ）アルカリ性条件下でエンド型プロテアーゼを作用させ、次いで（ｂ）中性〜酸性条件下でエキソ型プロテアーゼを作用させてなる酵素分解物に関する。更に本発明は、調味料原料に、酵素分解反応液中の塩濃度及びエタノール濃度をそれぞれ８〜１２ｗ／ｖ％及び３．５〜６ｖ／ｖ％、又はそれぞれ１ｗ／ｖ％以上８ｗ／ｖ％未満及び３．５〜１２ｖ／ｖ％に維持しつつ、２０〜４０℃で調味料原料に、（ａ）アルカリ性条件（塩濃度及びエタノール濃度が前者の場合はｐＨ８．５〜９．５、後者の場合はｐＨ９．５超〜１２）下でエンド型プロテアーゼを作用させ、次いで（ｂ）中性〜酸性条件下でエキソ型プロテアーゼを作用させることを特徴とする酵素分解物の製造方法に関する。更に本発明は、前述の酵素分解物を含有する食品、飲料又は調味料に関する。
本発明者らは特別な設備を必要とせずに、風味及び旨みの多い、ペプチドを多く含む酵素分解物の開発を目指した。その結果、２０〜４０℃において、酵素分解反応液中の塩濃度及びエタノール濃度をそれぞれ８〜１２ｗ／ｖ％及び３．５〜６ｖ／ｖ％、又はそれぞれ１ｗ／ｖ％以上８ｗ／ｖ％未満及び３．５〜１２ｖ／ｖ％に維持しつつ、調味料原料にアルカリ性条件下でエンド型プロテアーゼを作用させ、次いで中性〜酸性条件下でエキソ型プロテアーゼを作用させることにより、酵母、乳酸菌及びそれ以外の雑菌が生育せず、醤油香気及び加熱臭のない、風味及び旨みの多い、ペプチドを多く含むこく味の多い酵素分解物が得られることを見出した。更に該酵素分解物の製造方法において、酵素分解反応液中の塩濃度８〜１２ｗ／ｖ％及びエタノール濃度３．５〜６ｖ／ｖ％、又は塩濃度１ｗ／ｖ％以上８ｗ／ｖ％未満及びエタノール濃度３．５〜１２ｖ／ｖ％を維持しつつ、まずエンド型プロテアーゼをアルカリ性条件（塩濃度及びエタノール濃度が前者の場合はｐＨ８．５〜９．５、後者の場合はｐＨ９．５超〜１２）下で作用させることで、酵母、乳酸菌及びそれ以外の雑菌の汚染が防止しされ、かつ原料の分解率が向上すること、次いで、エキソ型プロテアーゼを中性〜酸性条件下で作用させることで、原料窒素回収率が向上し、醤油香気及び加熱臭のない、風味及び旨みの多い酵素分解物が得られることも見出し本発明の完成に至った。
また、本発明における酵素分解物の製造方法は、２０〜４０℃にて操作できるので、従来の方法では必要であった酵母、乳酸菌及びそれ以外の雑菌の汚染防止のための温度制御設備や厳密な作業管理を必要とせず、酵素分解反応液そのものの安定性も高いので、作業性に優れていると言う利点を有することも見出した。
更に、調味料原料として、穀類、畜肉類及び魚介類処理物等の産業廃棄物を用いて検討した結果、醤油香気及び加熱臭のない、旨み、こく味の良好な酵素分解物を得ることができることを見出した。

発明の詳細な説明
以下、本発明について具体的に説明する。
本発明における調味料原料とは、蛋白含有物及び／又は焼酎蒸留残渣をいう。
本発明における蛋白含有物とは、動物（魚介類を含む）または植物由来の蛋白を含有する物質をいう。蛋白含有物は、上記物質の処理物であってもよく、上記物質およびそれらの処理物から選択される２種以上の物質の混合物であってもよい。動物蛋白含有物には特に限定はないが、鶏肉、牛肉、豚肉等の畜肉が包含される。植物蛋白含有物にも特に限定はないが、小麦グルテン、コーングルテン、大豆粉末、脱脂大豆粉末等の穀類蛋白や胡麻粕、菜種粕、酒粕、豆腐粕（おから）等が、それぞれ例示される。また、それらの処理物とは上記のような動物蛋白含有物及び植物蛋白含有物の加工品、加工残渣、蛋白含有物の抽出物又は抽出残渣等を言う。蛋白含有物の処理物としては、小麦グルテンから加工される麩、コーンスターチの副産物であるコーングルテンミール（ＣＧＭ）、鰹節抽出物が例示される。従って、本発明の方法はＣＧＭの新規利用方法として有用である。
また、動物由来の蛋白含有物には魚介類処理物が含まれる。魚介類処理物とは、例えば、鰯、鯖、いか、蛸、海老、鰹、雑魚等の乾燥物又は焙乾物、又はそれらの残渣、抽出物、抽出残渣等を言い、魚介類の種類に特に限定はない。
ここで、魚介類の乾燥物は、加熱（又は煮熱）後、乾燥（天日干し又は乾燥機等で乾燥）した物、又は単に乾燥（天日干し又は乾燥機等で乾燥）した物であり、乾燥方法に特に限定はない。また、焙乾物は、乾燥と共に燻煙をかけた物であり、焙乾方法に特に限定はない。同じ燻煙をかけるにしても、燻製食品の場合には、燻煙をすることが主目的で乾燥は付随的なものであるが、かつお節を製造する際の焙乾は乾燥が主目的で燻煙が付随的なものというように異なっている〔だし・エキスの知識、第６１頁、著者太田静行、発行所（株）幸書房、１９９６年１２月２０日初版第１刷発行〕。
本発明に使用する焼酎蒸留残渣は、連続蒸留機（Ｃｏｎｔｉｎｕｏｕｓｓｔｉｌｌ）で蒸留する甲類焼酎の蒸留残渣、単式蒸留機（ｐｏｔｓｔｉｌｌ）で蒸留する乙類焼酎の蒸留残渣のいずれであっても良く、あるいは、ウオッカ、ウィスキーなどの蒸留酒の蒸留残渣であっても良いが、焼酎蒸留残渣中に含まれるエキス分を考慮に入れると、乙類焼酎蒸留残渣が好ましい。乙類焼酎蒸留残渣、特に減圧蒸留処理で得られた残渣中には、生きた酒精酵母菌体が相当量残っている。酒精酵母菌体は、その生育に適しない濃度の食塩及びエタノール含有条件で熟成することにより、自己消化が容易に起こり、菌体内蛋白、及び核酸分解物が遊離し、調味料に良好な香味及びこく味を加えることができる。乙類焼酎蒸留残渣としては、米焼酎、麦焼酎、芋焼酎、甘蔗焼酎、及び蕎麦焼酎等を生産する際に副生する蒸留廃液が例示される。廃液はそのままを使用しても良く、あるいは濃縮したものでも良い。またこれらの蒸留残渣は一種類又は数種類の混合物を使用しても良い。
また、本発明における原料の状態（原料粒度）については、特に限定はないが、例えば、微粉砕原料を使用した場合は、分解反応液中において大部分が浮遊状態で存在し、かくはんがほとんど不必要であり作業が容易となる。
本発明で使用するエタノールは、特に限定はなく、食品に使用できるものであれば良い。例えば、醸造用アルコール、又は、清酒、焼酎、ウオッカ、ウィスキー等のアルコール飲料がある。
本発明で使用する塩としては、通常は食塩、岩塩等の塩化ナトリウムを使用するが、塩化カリウム等の他の塩類も使用可能である。
本発明においてアルカリ性条件下で作用させるエンド型プロテアーゼは、アルカリ性条件下で高い酵素活性を持つものであれば特に限定はないが、アルカラーゼ〔ノボノルディスクバイオインダストリー（株）製〕、エスペラーゼ〔ノボノルディスクバイオインダストリー（株）製〕、オリエンターゼ２２ＢＦ〔阪急共栄物産（株）製〕、プロテアーゼＳ〔天野製薬（株）製〕等が例示される。また、本発明において中性〜酸性条件下で作用させるエキソ型プロテアーゼは、中性〜酸性条件下で高い酵素活性を持つものであれば特に限定はないが、フレーバーザイム〔ノボノルディスクバイオインダストリー（株）製〕、オリエンターゼＯＮＳ〔阪急共栄物産（株）製〕、プロテアーゼＡ〔天野製薬（株）製〕等が例示される。
本発明で使用するエンド型プロテアーゼ又はエキソ型プロテアーゼの形態としては、遊離の形又は担体に固定化された形でも良く、特に限定はない。例えば、担体に固定化されたエンド型プロテアーゼ又はエキソ型プロテアーゼ（以下、固定化酵素と称す）を用いる場合、実生産において酵素の使用量を飛躍的に減少させることができる。この場合、酵素分解反応液中の酵母、乳酸菌及びそれ以外の雑菌汚染が問題となるが、本発明においては、エタノール及び塩濃度を維持しつつ、アルカリ性又は中性〜酸性条件にて固定化酵素を作用させることができるので、２０〜４０℃においても効果的に酵母、乳酸菌及びそれ以外の雑菌汚染を防止できる。
本発明における酵素分解物は、ペプチド及びアミノ酸の混合物であるので、ペプチドおよび／またはアミノ酸を有効成分として含有するいずれかの組成物として使用することができる。この組成物をさらに精製して使用してもよい。組成物としては調味料が例示される。特に、本発明の酵素分解物は単独でこく味調味料として用いることができる。また、塩濃度１ｗ／ｖ％以上８％未満の酵素分解物は、スープ、出汁、たれ等の調味料の添加物及び栄養補助食品等の素材として用いることもできる。
本明細書において用いる用語「こく味調味料」は、ペプチドを比較的多量に含有し、味をなれさせ、こく味を効果的に付与する調味料をいう。アミノ酸を比較的多量に含有する醤油、ペプチド含量は比較的高いが特有の魚臭を有する魚醤油、魚肉エキスなどはこく味調味料に含まれない。本発明におけるこく味調味料において、ペプチド含有率は、式（１−ＦＮ／ＴＮ）で得られる値で表すことができる。ここで、ＴＮは可溶性画分中の全窒素のことであり、ＦＮは同じく可溶性画分中のホルモール窒素のことであり、それらの比であるＦＮ／ＴＮはアミノ酸含有率を表している。ＦＮ／ＴＮは例えば０．６〜０．１、好ましくは０．５〜０．２、より好ましくは０．４５〜０．２５である。
本明細書において用いる用語「こく味」とは、料理において、塩辛さ、すっぱさを抑える、味にまろみを付ける、濃厚感を増強するといったような総合的な作用をいう。
本明細書において用いる用語「酵母及び乳酸菌が生育しない条件」は、酵母及び乳酸菌が生育しないいずれかの条件をいう。５０〜６０℃のような高温での反応は得られる製品に加熱臭などが生じるので、特に調味料の製造には好ましくない。酵母及び乳酸菌の生育は、酵素分解物の一定量を取得し、これを適切な培地で培養し、その中に含まれる酵母及び乳酸菌の数を計数することによって測定することができる。例えば、酵母及び乳酸菌が生育しない条件は、酵素分解物１ｇあたり１０^４個以下の酵母及び乳酸菌を含有する条件である。
限定するものではないが、酵母は醤油酵母および酒精酵母を包含し、乳酸菌は醤油乳酸菌を包含する。
１つの実施態様において、酵母及び乳酸菌が生育しない条件は、酵素分解反応液中の塩濃度及びエタノール濃度を適切に調整することによって提供される。
次に、本発明におけるエタノール濃度と塩濃度は、例えば、２０〜４０℃において酵素分解反応液中のエンド型プロテアーゼ又はエキソ型プロテアーゼ活性を阻害せずに、酵母、及び乳酸菌の増殖を阻止するような条件であれば、該条件を達成する手段に特に限定はないが、好ましくは、酵素分解反応液中のエタノール及び塩濃度を調整することによって達成する。なお、酵素分解反応液中のエタノール及び塩濃度を調整するに当っては、例えば、原料由来の持込み分がある場合には、エタノール及び塩を補正して添加する必要がある。
酵素分解反応液中のエタノール濃度が３．５ｖ／ｖ％未満又は塩濃度が８ｗ／ｖ％未満では酵素分解反応液中に酵母及び乳酸菌が生育し、熟成後の酵素分解反応液は腐敗してしまい、また、エタノール濃度が６ｖ／ｖ％又は塩濃度が１２ｗ／ｖ％を超えた場合は、酵母及び乳酸菌の生育又は腐敗は無いが、酵素分解反応液中のエンド型プロテアーゼ又はエキソ型プロテアーゼ活性が阻害を受け、その結果、原料中窒素の利用率が低くなる。したがって、本発明のこく味調味料を得るためには、酵素分解反応液中のエタノール濃度３．５〜６ｖ／ｖ％及び塩濃度８〜１２ｗ／ｖ％にすることが望ましい。なお、酵素分解反応液中のエタノールは熟成中に一部が蒸散するので、必要に応じて補充することが望ましい。
前述のごとく、塩濃度８ｗ／ｖ％未満では通常、酵素分解反応液中の酵母及び乳酸菌が生育し、酵素分解反応液は腐敗してしまうが、酵素反応におけるアルカリ性条件がｐＨ９．５超〜１２で、かつ、エタノール濃度３．５〜１２ｖ／ｖ％に設定した場合は、この限りではない。すなわち、塩濃度が１ｗ／ｖ％以上８ｗ／ｖ％未満の酵素分解物を得る場合は、ｐＨ及びエタノール濃度を上記の範囲に設定することで腐敗を防止できる。エタノール濃度が６ｖ／ｖ％を越えた場合、酵素分解反応液中のエンド型プロテアーゼ又はエキソ型プロテアーゼ活性が阻害を受けるが、その場合は、使用する酵素量を増やすか反応時間を延長することで対応できる。
本発明における反応温度は、例えば、２０〜４０℃であり、該範囲中に収まるのであれば温度コントロールをしても良いし、気候により成り行きに任せても良い。上記のように、５０〜６０℃のような高温での反応は得られる製品に加熱臭などが生じるので、特に調味料の製造には好ましくない。
本発明の酵素分解物の製造は、例えば以下のように行う。すなわち、所定量の調味料原料、エタノール及び食塩を混合した分解反応液を、まずアルカリ性条件下でエンド型プロテアーゼを作用させ、次いで中性〜酸性条件下でエキソ型プロテアーゼを作用させる。反応温度は、２０〜４０℃、エンド型プロテアーゼ及びエキソ型プロテアーゼの酵素活性を考慮に入れると好ましくは２５〜３５℃にて、通常１〜８週間分解、熟成を行う。
本発明にて使用するエンド型プロテアーゼの添加量は、好ましくは原料乾物ｇ当り２００単位〜５０００単位であればよく、特に好ましくは、８００単位〜３０００単位であればよい。なお、ここでいうプロテアーゼの活性は、当該酵素をカゼインを基質として、１分間に１μｇのチロシンに相当するＴＣＡ可溶性のフォリン試薬呈色物質を生成する酵素量を１単位（Ｕ）とする。
また、本発明にて使用するエキソ型プロテアーゼの添加量は、好ましくは原料乾物ｇ当り２単位〜５０単位であればよく、特に好ましくは、５単位〜２０単位であればよい。なお、ここでいうプロテアーゼの活性は、当該酵素をＬ−ロイシン−ｐ−ニトロアニリドを基質として、１分間に１マイクロモルのｐ−ニトロアニリドを遊離する酵素量を１単位（Ｕ）とする。
また、本発明において使用する酵素は、微生物の培養物又はその加工品でも良い。例えば、アスペルギルス（Ａｓｐｅｒｇｉｌｌｕｓ）属菌、バチルス（Ｂａｃｉｌｌｕｓ）属菌の液体培地培養物又は固体培地培養物、特に米麹、豆麹、麦麹、小麦ふすま麹、醤油麹等でも良い。
本発明における分解反応液のｐＨは、酵素の至適ｐＨ範囲に調整すれば特に限定はないが、例えば、一般的なこく味調味料の製造においては、最初にアルカリ性条件下、好ましくは、ｐＨ８．５〜９．５でエンド型プロテアーゼを作用させ、次いで中性〜酸性にｐＨを変更して、エキソ型プロテアーゼを作用させるのが好ましく、中性〜酸性条件がｐＨ５．５〜６．５であることが特に好ましい。本明細書において、アルカリ性、中性、酸性はこれらの用語が一般的に使用される場合と同じ意味を表す。すなわち、アルカリ性はｐＨが７より大きいことをいい、中性はｐＨが７であることをいい、酸性はｐＨが７より小さいことをいう。まず、最初の酵素反応をアルカリ側で行うことで、腐敗菌の混入を防ぎながら、原料の膨潤と可溶化が進み、酵素の反応性が高まる。また、エンド型プロテアーゼを用いることで、蛋白質の可溶化、ペプチド化が促進される。次いで、中性〜酸性条件下でエキソ型プロテアーゼを作用させることで、旨味成分の生成が促進される。その結果として、原料利用率が高く、旨味、こく味の多い調味料の製造が可能となる。更に、従来の加熱しながらアルカリ性条件下で蛋白質分解酵素を作用させる方法では、加熱臭及び脱アミノ化反応によるアンモニア臭やアルカリ臭等が発生するが、本発明の条件においては、常温下で蛋白質分解酵素を作用させることができるので、そのような臭気の発生が見られない。
また、例えば、塩濃度が１ｗ／ｖ％以上８ｗ／ｖ％未満の酵素分解物の製造においては、まず、ｐＨ９．５超〜１２でエンド型プロテアーゼを作用させ、次いで中性〜酸性条件下でエキソ型プロテアーゼを作用させればよい。
このようにして得られた調味料又は酵素分解物はこのままでも各種用途に使用可能であるが、必要に応じて公知処理方法を実施することで更に高品質の製品とすることができる。処理手段としては、１．使用した酵素を失活させるための加熱処理（例えば、８０℃、３０分間）、２．固形分を除去するための遠心分離、ろ過、圧搾処理、３．脱臭、脱色のための活性炭処理、４．濃縮処理操作、及び５．スプレードライ等による粉末化等を単独でも又は複数組合せても良い。
また、本発明により前述の酵素分解物を含有することを特徴とする食品、飲料又は調味料が提供される。本発明の食品、飲料又は調味料には、必要に応じ、甘味料、酸味料、香料等を添加しても良く、更にエタノール及び／又は炭酸ガスを添加し、エタノール含有飲料又は炭酸ガス含有飲料としても良い。なお、本発明の食品、飲料又は調味料でいう「含有」の語は、含有、添加、希釈の意を含むものであり、含有とは食品、飲料又は調味料中に本発明により得られる酵素分解物が含まれるという態様を、添加とは食品、飲料又は調味料の原料に、該酵素分解物を添加するという態様を、希釈とは該酵素分解物を食品、飲料又は調味料の原料で希釈するという態様をいうものである。
本発明の食品、飲料又は調味料とは、前述の酵素分解物を含有すれば良く、特に限定はされないが、例えば、麺類、パン類等の穀物加工品、野菜飲料、果実飲料、漬け物類等の野菜・果実加工品、リキュール類等のアルコール飲料、だし、たれ等の調味料が挙げられる。
以上説明したごとく、本発明の酵素分解物は、醤油香気及び加熱臭を発生しない、魚醤油や醤油にない、風味、旨み、こく味を有する新規の酵素分解物である。また、本発明の酵素分解物を食品又は飲料に使用することにより、塩なれ感、酢なれ感、まろやかさの付与、味の伸びなど熟成した味わいをもたせることができる。
以下、本発明を実施例によって更に具体的に説明するが、本発明はこれらの実施例に限定されるものではない。

実施例
実施例１
鰹節粉末〔（株）マルモ製、１００ｍｅｓｈｐａｓｓＴＮ１３％〕８０ｇ（乾物重量７５ｇ）、食塩５０ｇ、エタノール２５ｍｌ、水４００ｍｌを混合して、食塩及びエタノール終濃度がそれぞれ１０ｗ／ｖ％及び５ｖ／ｖ％になった混合液を用意した。次に、該混合液のｐＨを８．５に調整したのちアルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３ｍｌ添加して３０℃にて静置した。静置中の混合液は、原料が微粉末であるため大部分が液中に浮遊しており、かくはんはほとんど不要であった。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて８．５になるように調整した。４週間後、該混合液のｐＨを６．５に調整した後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８ｍｌを添加して、１週間静置し、反応を終了した。次に、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．４％、ＦＮ０．４％、ＮａＣｌ１０．０％、ＥｔＯＨ５％、ｐＨ５．５、Ｂｒｉｘ１９．６、ＯＤ_４３０５．１の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた調味料が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

参考例１
干麩８４ｇ（乾物重量７８ｇ）、食塩５０ｇ、エタノール２５ｍｌ、水３７５ｍｌを混合し、ｐＨを８．５に調整したのちアルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３ｍｌ添加して３０℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて８．５になるように調整した。１週間後、該混合液のｐＨを６．０に調整した後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８ｍｌを添加して、１週間静置し、反応を終了した。次に、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．６％、ＦＮ０．４５％、ＮａＣｌ１０．０％、ＥｔＯＨ５％、ｐＨ５．８、Ｂｒｉｘ２８．１、ＯＤ_４３０３．１の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた調味料が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

実施例２
鰹節抽出残渣（ＴＮ６％）１５０ｇ（乾物重量７５ｇ）、食塩３４ｇ、エタノール１７ｍｌ、水１７０ｍｌを混合し、ｐＨを８．５に調整したのちアルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３ｍｌ添加して３０℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて８．５になるように調整した。４週間後、該混合液のｐＨを５．５に調整した後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８ｍｌを添加して、１週間静置し、反応を終了した。次に、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．９％、ＦＮ０．８％、ＮａＣｌ１０．０％、ＥｔＯＨ５％、ｐＨ５．４、Ｂｒｉｘ２５．４、ＯＤ_４３０５．９の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた調味料が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

参考例２
コーングルテンミール（ＴＮ１１．９％）８６ｇ、食塩５０ｇ、エタノール２５ｍｌ、水３７３ｍｌを混合し、ｐＨを９．０に調整した後、アルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３３ｍｌ添加して３０℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて９．０になるように調整した。４週間後、該混合液のｐＨを５．０に調整した後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８２ｍｌを添加して、２週間静置し、反応を終了した。次に、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．７％、ＦＮ０．５１％、ＮａＣｌ１０．５％、ＥｔＯＨ４．５％、Ｂｒｉｘ２４．０、ＯＤ_４３０４．５の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた調味料が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

実施例３
鰹節抽出残渣（ＴＮ６．９％）１８１．２ｇ（乾物重量８７．０ｇ）、食塩３０ｇ、エタノール２５ｍｌ、水２８２．９ｍｌを混合し、ｐＨを１０．０に調整したのちアルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３３ｍｌ添加して、合計５００ｍｌを３０℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて１０．０になるように調整した。４週間後、該混合液のｐＨを５．５に調整した後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８ｍｌを添加して、２週間静置し、反応を終了した。次に、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．６％、ＦＮ０．４８％、ＮａＣｌ６．０％、ＥｔＯＨ５％、ｐＨ５．４、Ｂｒｉｘ２０．５、ＯＤ_４３０３．８の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた酵素分解物が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

実施例４
鰹節抽出残渣（ＴＮ６．９％）１８１．２ｇ（乾物重量８７．０ｇ）、食塩２５ｇ、エタノール３０ｍｌ、水２８７．９ｍｌを混合し、ｐＨを９．６に調整したのちアルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．３３ｍｌ添加して、合計５００ｍｌを３０℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは時々ＮａＯＨ溶液（４０ｗ／ｖ％）を用いて９．６になるように調整した。５週間後、該混合液のｐＨを５．５に調整したした後、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕０．８ｍｌを添加して更に１週間静置した。次いで、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ１．７％、ＦＮ０．４９％、ＮａＣｌ５．１％、ＥｔＯＨ５．９％、ｐＨ５．３、Ｂｒｉｘ１９．６、ＯＤ_４３０３．９の醤油香気及び加熱臭のない、風味、旨み、こく味に優れた酵素分解物が得られた。また、分解反応中の雑菌による腐敗は見られなかった。

実施例５
麦焼酎蒸留廃液（ＴＮ０．７１％、Ｂｒｉｘ２４．８）６００ｍｌに干麩（ＴＮ１１．９％）１３５ｇ、エタノール４０ｍｌ、食塩８０ｇを加え、ｐＨを８．６に調製したのち、アルカラーゼ２．４Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．４ｍｌ添加して、合計８００ｍｌを３５℃にて静置した。静置中の混合液は、１日に１回かくはんを行った。また、混合液のｐＨは、時々ＮａＯＨ溶液（４０ｗ／ｖ％）で８．６になるように調整した。３週間後に、ｐＨを６．５に調節したのち、フレーバーザイム１０００Ｌ〔ノボノルディスクバイオインダストリー（株）製〕を０．９ｍｌ添加して更に１週間静置した。次いで、固液分離し、８０℃、３０分間加熱して酵素を失活させた後、ろ過をして調味料を得た。一般分析値はＴＮ２．２％、ＦＮ０．８％、ＮａＣｌ１０．５％、ＥｔＯＨ５．５％、ｐＨ５．８、Ｂｒｉｘ２２．０１９．６、ＯＤ_４３０５．６で、醤油香気及び加熱臭のない、風味、こく味に優れたものであった。また、分解反応中の雑菌による腐敗は見られなかった。

産業上の利用の可能性
本発明により、風味、旨み、こく味の良い酵素分解物を提供することができる。また、本発明により、節抽出残渣、焼酎蒸留残渣やおから等の副産物の有効利用方法を提供できる。更に、本発明においては、酵素分解槽等の特別な装置を必要とせず、経済的に安価な酵素分解物を製造することが可能である。 Technical field
  The present invention relates to an enzyme degradation product obtained by allowing a proteolytic enzyme to act on a seasoning material under specific conditions, a seasoning comprising the enzyme degradation product, and a method for producing the same.

Background art
  Conventionally, various seasonings have been manufactured by hydrolyzing the raw material with hydrochloric acid or the like, but since harmful chlorine compounds such as MCP and DCP are also by-produced, there has been an interest in food safety in recent years. With the rise, attention has been focused on producing seasonings by enzymatic degradation. For example, the fish is heated to a temperature of 60 ° C. or higher to inactivate autolytic enzymes contained in the fish body, and then decomposed by adding an alkali-resistant proteolytic enzyme at 50 to 60 ° C. and pH 9 to 10, followed by pH Is adjusted to 5-6, another acid-resistant proteolytic enzyme is added, and the seasoning is also produced at 50-60 ° C. (Japanese Examined Patent Publication No. 53-31935). Manufactures liquid seasonings characterized by neutralizing type proteases to solubilize proteins and then acidifying exotype proteases to decompose bitter components and increase umami components Method (Japanese Patent Laid-Open No. 5-308922), a method of producing a seasoning by enzymatically degrading wheat gluten, animal protein (gelatin) and the like under 50-60 ° C. conditions (food and development, 31, 2, 20 22 pp., 1996) there is. In these conventional enzymatic decompositions, the temperature in the enzymatic decomposition reaction solution must be set to a high temperature of 50 to 60 ° C. in order to prevent contamination with various bacteria in the enzymatic decomposition reaction solution, and as a result, it is obtained. There was a problem that the product was unavoidably heated. Further, the exo-type protease generally used is derived from filamentous fungi, has low heat resistance, and when the temperature in the enzymatic decomposition reaction solution exceeds 55 ° C., the enzyme activity is gradually deactivated. Therefore, in actual production, an enzyme decomposition reaction tank equipped with temperature control, stirring ability, etc. is necessary, and there is a problem that considerable capital investment, production management personnel and maintenance costs are required.
  On the other hand, in the method for producing fish soy sauce, raw fish and shellfish with an inorganic salt concentration of 3 to 12% (w / w) in the presence of proteolytic enzyme and ethanol corresponding to an ethanol concentration of 0.5 to 5% (v / v) Has been disclosed (Japanese Patent Application Laid-Open No. 3-254660). However, this method uses common food enzymes in a common way for the production of fish soy sauce (ie in the presence of ethanol, inorganic salts, etc.). The conditions in this method are not suitable for the use of an effective enzyme that takes advantage of the properties of the enzyme (including the optimum pH), and thus have the disadvantage that the raw material utilization rate is low. In particular, development of technology for utilizing industrial waste with a high raw material utilization rate has been demanded from the current situation where development of effective utilization technology for extraction residues discharged from stock soup manufacturing companies and the like is required.
  Corn gluten meal (CGM) is a resource that is produced in large quantities (approximately 200,000 tons per year in Japan) as a by-product of corn starch and contains a large amount of protein. However, since CGM is difficult to be decomposed, it is currently used only for livestock feed. Therefore, development of a more effective method for using CGM is desired.

Object of the invention
  A first object of the present invention is to provide an enzyme degradation product obtained by enzymatic degradation of a seasoning material using a simple facility that does not require special facilities. A second object is to provide a first object. It is in providing the manufacturing method of an enzyme degradation product.

Summary of the Invention
  Briefly, the present invention is such that (a) an endo-type protease is allowed to act under alkaline conditions under conditions where yeast and lactic acid bacteria do not grow, and then (b) exo under under neutral to acidic conditions. The present invention relates to an enzymatic degradation product obtained by allowing a type protease to act. Furthermore, the present invention provides the seasoning material with a salt concentration and an ethanol concentration of 8 to 12 w / v% and 3.5 to 6 v / v%, or 1 w / v% or more and 8 w / v%, respectively, in the enzyme decomposition reaction solution. Less than and 3.5 to 12 v / v% while maintaining the seasoning raw material at 20 to 40 ° C., (a) alkaline conditions (pH 8.5 to 9.5 when the salt concentration and ethanol concentration are the former, the latter In the case of the above, the present invention relates to a method for producing an enzymatic degradation product, characterized in that an endo-type protease is allowed to act under pH 9.5 to 12) and then (b) an exo-type protease is allowed to act under neutral to acidic conditions. Furthermore, this invention relates to the foodstuff, drink, or seasoning containing the above-mentioned enzyme degradation product.
  The present inventors aimed to develop an enzymatic degradation product containing a large amount of peptides and having a high flavor and taste without requiring special equipment. As a result, at 20 to 40 ° C., the salt concentration and ethanol concentration in the enzymatic decomposition reaction solution were 8 to 12 w / v% and 3.5 to 6 v / v%, respectively, or 1 w / v% or more and less than 8 w / v%, respectively. And 3.5 to 12% v / v, while allowing the seasoning raw material to act with an endo-type protease under alkaline conditions and then with an exo-type protease under neutral to acidic conditions, yeast, lactic acid bacteria and It has been found that other miscellaneous bacteria do not grow, and there is no soy sauce flavor and heating odor, and there is a rich flavor and umami-rich enzyme-decomposed product containing a large amount of peptide. Furthermore, in the method for producing the enzyme degradation product, the salt concentration in the enzyme decomposition reaction solution is 8 to 12 w / v% and the ethanol concentration is 3.5 to 6 v / v%, or the salt concentration is 1 w / v% or more and less than 8 w / v%, and While maintaining the ethanol concentration of 3.5 to 12 v / v%, the endo-type protease is first subjected to alkaline conditions (pH 8.5 to 9.5 when the salt concentration and ethanol concentration are the former, and over pH 9.5 when the latter is 12) By making it act below, contamination of yeast, lactic acid bacteria and other miscellaneous bacteria is prevented and the decomposition rate of the raw material is improved, and then exo-type protease is allowed to act under neutral to acidic conditions. Thus, it was found that the raw material nitrogen recovery rate was improved, and an enzyme-decomposed product having a high flavor and taste was obtained without soy sauce flavor and heating odor, and the present invention was completed.
  Moreover, since the manufacturing method of the enzyme degradation product in this invention can be operated at 20-40 degreeC, the temperature control equipment for the contamination | pollution prevention of yeast, lactic acid bacteria, and other miscellaneous bacteria which were required in the conventional method, and strict The present inventors have also found that there is an advantage that the workability is excellent because the operation of the enzyme decomposition reaction solution itself is high and the workability is not required.
  Furthermore, as a result of investigation using industrial waste such as processed cereals, livestock meats and fishery products as seasoning raw materials, it is possible to obtain an enzyme degradation product with good taste and richness without soy sauce aroma and heated odor. I found out that I can do it.

Detailed Description of the Invention
  Hereinafter, the present invention will be specifically described.
  The seasoning raw material in the present invention refers to a protein-containing material and / or a shochu distillation residue.
  The protein-containing material in the present invention refers to a substance containing protein derived from animals (including seafood) or plants. The protein-containing product may be a processed product of the above-mentioned substance, or may be a mixture of two or more kinds of substances selected from the above-mentioned substance and those processed products. The animal protein-containing material is not particularly limited, but includes livestock such as chicken, beef and pork. The plant protein-containing material is not particularly limited, and examples thereof include cereal proteins such as wheat gluten, corn gluten, soybean powder, and defatted soybean powder, sesame meal, rapeseed meal, sake lees, and tofu meal (okara). . In addition, these processed products refer to processed products of animal protein-containing products and plant protein-containing products, processed residues, extracts of protein-containing products, or extracted residues. Examples of the processed protein-containing product include cocoon processed from wheat gluten, corn starch corn gluten meal (CGM), and koji extract. Therefore, the method of the present invention is useful as a novel method of using CGM.
  Animal-derived protein-containing products include processed seafood. Processed seafood refers to, for example, dried or roasted products such as salmon, salmon, squid, salmon, shrimp, salmon, miscellaneous fish, etc., or their residues, extracts, extraction residues, etc. There is no limitation.
  Here, the dried fishery product is a product that has been heated (or boiled) and then dried (dried by sun-drying or drying) or simply dried (dried by sun-drying or drying). There is no particular limitation on the drying method. In addition, the dried product is a product that has been smoked with drying, and the drying method is not particularly limited. Even if the same smoke is applied, in the case of smoked foods, the main purpose is to smoke, and drying is incidental, but the drying of bonito is the main purpose of drying. Smoke is an incidental thing (knowledge of dashi and extract, page 61, author Shizuyuki Ota, publisher Shokobo Co., Ltd., first edition published on December 20, 1996).
  The shochu distillation residue used in the present invention may be either a distillation residue of the first class shochu distilled by a continuous still or a second residue of a second class shochu distilled by a single type still (pot still). Or, it may be a distillation residue of distilled liquor such as vodka, whiskey, etc., but taking into consideration the extract contained in the shochu distillation residue, the oyster shochu distillation residue is preferred. A considerable amount of vigorous alcoholic yeast cells remain in the sesame shochu distillation residue, particularly the residue obtained by the vacuum distillation treatment. By aging under conditions containing salt and ethanol at a concentration that is not suitable for growth, the sake yeast cells are easily self-digested, releasing intracellular protein and nucleic acid degradation products, and having a good flavor and seasoning. Kokumi can be added. Examples of the sesame shochu distillation residue include distillation waste liquid by-produced when producing rice shochu, barley shochu, shochu shochu, sweet potato shochu, soba shochu, and the like. The waste liquid may be used as it is or may be concentrated. These distillation residues may use one kind or a mixture of several kinds.
  In addition, the state of the raw material (raw material particle size) in the present invention is not particularly limited. For example, when a finely pulverized raw material is used, most of it is present in a floating state in the decomposition reaction solution, and stirring is almost impossible. It is necessary and easy to work.
  The ethanol used in the present invention is not particularly limited as long as it can be used for food. For example, there are alcohol for brewing or alcoholic beverages such as sake, shochu, vodka, whiskey.
  As the salt used in the present invention, sodium chloride such as salt and rock salt is usually used, but other salts such as potassium chloride can also be used.
  In the present invention, the endo-type protease that acts under alkaline conditions is not particularly limited as long as it has a high enzyme activity under alkaline conditions, but Alcalase [manufactured by Novo Nordisk Bio Industry Co., Ltd.], esperase [Novo Norol] Disc Bio Industry Co., Ltd.], Orientase 22BF (Hankyu Kyoei Bussan Co., Ltd.), Protease S (Amano Pharmaceutical Co., Ltd.) and the like are exemplified. In the present invention, the exo-type protease that acts under neutral to acidic conditions is not particularly limited as long as it has a high enzyme activity under neutral to acidic conditions, but flavorzyme [Novonordisk bioindustry ( Co., Ltd.], Orientase ONS [manufactured by Hankyu Kyoei Bussan Co., Ltd.], protease A [manufactured by Amano Pharmaceutical Co., Ltd.], and the like.
  The form of the endo-type protease or exo-type protease used in the present invention may be a free form or a form immobilized on a carrier, and is not particularly limited. For example, when an endo-type protease or exo-type protease (hereinafter referred to as an immobilized enzyme) immobilized on a carrier is used, the amount of enzyme used can be drastically reduced in actual production. In this case, contamination of yeast, lactic acid bacteria and other miscellaneous bacteria in the enzymatic decomposition reaction solution becomes a problem. In the present invention, the immobilized enzyme is maintained under alkaline or neutral to acidic conditions while maintaining ethanol and salt concentrations. Therefore, it is possible to effectively prevent yeast, lactic acid bacteria and other miscellaneous bacteria from contaminating even at 20 to 40 ° C.
  Since the enzymatic degradation product in the present invention is a mixture of peptides and amino acids, it can be used as any composition containing peptides and / or amino acids as active ingredients. This composition may be used after further purification. A seasoning is illustrated as a composition. In particular, the enzyme degradation product of the present invention can be used alone as a rich seasoning. In addition, an enzyme degradation product having a salt concentration of 1 w / v% or more and less than 8% can also be used as an additive for seasonings such as soup, soup stock, and sauce, and as a material for nutritional supplements.
  The term “kokumi seasoning” used in the present specification refers to a seasoning that contains a relatively large amount of peptide, makes it taste good, and effectively imparts kokumi. Soy sauce containing a relatively large amount of amino acids, fish soy sauce having a relatively high peptide content but having a characteristic fish odor, and fish meat extract are not included in the rich seasoning. In the rich seasoning in the present invention, the peptide content can be represented by a value obtained by the formula (1-FN / TN). Here, TN is the total nitrogen in the soluble fraction, FN is also the formol nitrogen in the soluble fraction, and the ratio FN / TN represents the amino acid content. FN / TN is, for example, 0.6 to 0.1, preferably 0.5 to 0.2, and more preferably 0.45 to 0.25.
  The term “kokumi” used in the present specification refers to a comprehensive action such as suppressing saltiness and sourness, adding a mellow taste, and enhancing a rich feeling in cooking.
  The term “conditions in which yeast and lactic acid bacteria do not grow” used in the present specification refers to any condition in which yeast and lactic acid bacteria do not grow. A reaction at a high temperature such as 50 to 60 ° C. is not preferable for the production of a seasoning because a heated odor or the like is generated in the resulting product. The growth of yeast and lactic acid bacteria can be measured by obtaining a certain amount of the enzyme degradation product, culturing it in an appropriate medium, and counting the number of yeast and lactic acid bacteria contained therein. For example, the conditions under which yeast and lactic acid bacteria do not grow are 10 per 1 g of enzymatic degradation product.⁴This is a condition containing no more than one yeast and lactic acid bacteria.
  Although not limited, yeast includes soy sauce yeast and sake spirit yeast, and lactic acid bacteria include soy sauce lactic acid bacteria.
  In one embodiment, conditions under which yeast and lactic acid bacteria do not grow are provided by appropriately adjusting the salt concentration and ethanol concentration in the enzymatic degradation reaction solution.
  Next, the ethanol concentration and the salt concentration in the present invention, for example, prevent the growth of yeast and lactic acid bacteria without inhibiting the endo-type protease or exo-type protease activity in the enzymatic degradation reaction solution at 20 to 40 ° C. There are no particular limitations on the means for achieving these conditions, but it is preferably achieved by adjusting the ethanol and salt concentrations in the enzymatic decomposition reaction solution. In addition, when adjusting the ethanol and salt concentration in the enzymatic decomposition reaction solution, for example, when there is a carry-in derived from the raw material, it is necessary to correct and add the ethanol and salt.
  If the ethanol concentration in the enzyme decomposition reaction solution is less than 3.5 v / v% or the salt concentration is less than 8 w / v%, yeast and lactic acid bacteria grow in the enzyme decomposition reaction solution, and the enzyme decomposition reaction solution after ripening is spoiled. In addition, when the ethanol concentration exceeds 6 v / v% or the salt concentration exceeds 12 w / v%, there is no growth or decay of yeast and lactic acid bacteria, but the endo-type protease or exo-type protease activity in the enzymatic degradation reaction solution As a result, the utilization rate of nitrogen in the raw material is lowered. Therefore, in order to obtain the kokumi seasoning of the present invention, it is desirable that the ethanol concentration in the enzymatic decomposition reaction solution is 3.5 to 6 v / v% and the salt concentration is 8 to 12 w / v%. In addition, since ethanol in the enzymatic decomposition reaction solution partially evaporates during aging, it is desirable to replenish as necessary.
  As described above, when the salt concentration is less than 8 w / v%, yeast and lactic acid bacteria in the enzyme decomposition reaction solution usually grow and the enzyme decomposition reaction solution rots, but the alkaline condition in the enzyme reaction is more than pH 9.5 to 12 However, this is not the case when the ethanol concentration is set to 3.5 to 12 v / v%. That is, when obtaining an enzyme degradation product having a salt concentration of 1 w / v% or more and less than 8 w / v%, it is possible to prevent spoilage by setting the pH and ethanol concentration within the above ranges. When the ethanol concentration exceeds 6v / v%, endo-type protease or exo-type protease activity in the enzymatic degradation reaction solution is inhibited. In this case, increase the amount of enzyme used or extend the reaction time. Yes.
  The reaction temperature in the present invention is, for example, 20 to 40 ° C., and the temperature may be controlled as long as it falls within the range, or may be left to the future depending on the climate. As described above, a reaction at a high temperature such as 50 to 60 ° C. is not preferable for the production of seasonings because a heated odor or the like is generated in the resulting product.
  The enzymatic degradation product of the present invention is produced, for example, as follows. That is, a decomposition reaction solution obtained by mixing a predetermined amount of a seasoning raw material, ethanol and salt is first subjected to endo-type protease under alkaline conditions, and then allowed to act under neutral to acidic conditions. When the reaction temperature is 20 to 40 ° C. and the enzyme activities of endo-type protease and exo-type protease are taken into consideration, the reaction temperature is preferably 25 to 35 ° C., and the decomposition and aging are usually performed for 1 to 8 weeks.
  The addition amount of the endo-type protease used in the present invention is preferably 200 units to 5000 units, and particularly preferably 800 units to 3000 units, per g of raw material dry matter. The protease activity referred to here is defined as the amount of enzyme that produces a TCA-soluble forin reagent coloring substance corresponding to 1 μg of tyrosine per minute with the enzyme as casein as a substrate.
  Moreover, the addition amount of the exo-type protease used in the present invention is preferably 2 units to 50 units, and particularly preferably 5 units to 20 units, per g dry material. In addition, the activity of protease here makes the amount of enzyme which liberates 1 micromol p-nitroanilide in 1 minute by making the said enzyme L-leucine-p-nitroanilide into a substrate as 1 unit (U).
  The enzyme used in the present invention may be a culture of microorganisms or a processed product thereof. For example, a liquid medium culture or solid medium culture of Aspergillus or Bacillus, particularly rice bran, soybean cake, wheat straw, wheat bran or soy sauce cake may be used.
  The pH of the decomposition reaction solution in the present invention is not particularly limited as long as it is adjusted to the optimum pH range of the enzyme. For example, in the production of general kokumi seasonings, first, the alkaline reaction conditions, preferably pH 8 It is preferable that endo-protease is allowed to act at .5 to 9.5, and then pH is changed from neutral to acidic to cause exo-type protease to act, and neutral to acidic conditions are at pH 5.5 to 6.5. It is particularly preferred. In the present specification, alkaline, neutral, and acidic represent the same meaning as when these terms are generally used. That is, alkaline means that the pH is greater than 7, neutral means that the pH is 7, and acidic means that the pH is less than 7. First, by performing the first enzyme reaction on the alkali side, the raw material swells and solubilizes while preventing the entry of spoilage bacteria, and the reactivity of the enzyme increases. In addition, by using an endo-type protease, protein solubilization and peptide formation are promoted. Subsequently, the production | generation of an umami | taste component is accelerated | stimulated by making exo-type protease act under neutral-acidic conditions. As a result, it is possible to produce a seasoning with a high raw material utilization rate and a high umami and rich taste. Furthermore, in the conventional method in which a proteolytic enzyme is allowed to act under alkaline conditions while heating, ammonia odor or alkali odor due to heating odor and deamination reaction is generated. Since a degrading enzyme can be made to act, generation of such an odor is not seen.
  For example, in the production of an enzyme degradation product having a salt concentration of 1 w / v% or more and less than 8 w / v%, first, an endo-type protease is allowed to act at a pH of more than 9.5 to 12, and then under neutral to acidic conditions. An exo-type protease may be allowed to act.
  The seasoning or enzyme degradation product obtained in this way can be used for various purposes as it is, but it can be made into a higher quality product by carrying out known processing methods as necessary. As processing means,1.Heat treatment (for example, 80 ° C., 30 minutes) to inactivate the used enzyme,2.Centrifugation, filtration, squeezing to remove solids,3.Activated carbon treatment for deodorization and decolorization,4).Concentration processing operation, and5.Powdering by spray drying or the like may be used alone or in combination.
  The present invention also provides a food, beverage or seasoning characterized by containing the aforementioned enzyme degradation product. The food, beverage or seasoning of the present invention may contain a sweetener, a sour agent, a fragrance, etc., if necessary, and further ethanol and / or carbon dioxide is added to the ethanol-containing beverage or carbon dioxide-containing beverage. It is also good. The term “containing” in the food, beverage or seasoning of the present invention includes inclusion, addition and dilution, and the term “containing” means an enzyme obtained by the present invention in food, beverage or seasoning. A mode in which a degradation product is contained, an addition is a mode in which the enzyme degradation product is added to a raw material for food, beverage or seasoning, and a mode in which dilution is a raw material for food, beverage or seasoning. It means an aspect of dilution.
  The food, beverage or seasoning of the present invention is not particularly limited as long as it contains the above-mentioned enzyme degradation product, for example, processed products such as noodles, breads, vegetable beverages, fruit beverages, pickles, etc. Processed fruits and vegetables, alcoholic beverages such as liqueurs, and seasonings such as dashi and sauce.
  As described above, the enzymatic degradation product of the present invention is a novel enzymatic degradation product that does not generate soy sauce flavor and heated odor, and does not exist in fish soy sauce or soy sauce, and has a flavor, umami, and richness. In addition, by using the enzyme degradation product of the present invention in foods or beverages, it is possible to have an aged taste such as a salty feeling, a vinegared feeling, a mellowness, and a taste increase.
  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

Example
Example 1
  80 g of dried bonito powder (manufactured by Marumo Corporation, 100 mesh pass TN 13%), 50 g of salt, 25 ml of ethanol and 400 ml of water are mixed, and the final concentrations of salt and ethanol are 10 w / v% and 5 v / v, respectively. % Mixed solution was prepared. Next, after adjusting the pH of the mixed solution to 8.5, 0.3 ml of Alcalase 2.4L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] was added and allowed to stand at 30 ° C. Most of the liquid mixture during standing was suspended in the liquid because the raw material was fine powder, and stirring was almost unnecessary. Further, the pH of the mixed solution was sometimes adjusted to 8.5 using a NaOH solution (40 w / v%). After 4 weeks, the pH of the mixed solution was adjusted to 6.5, and then 0.8 mL of Flavorzyme 1000 L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] was added and left to stand for 1 week to complete the reaction. . Next, after heating at 80 degreeC for 30 minutes and deactivating an enzyme, it filtered and obtained the seasoning. General analysis values are TN 1.4%, FN 0.4%, NaCl 10.0%, EtOH 5%, pH 5.5, Brix 19.6, OD₄₃₀  A seasoning excellent in flavor, umami and kokumi without 5.1 soy sauce flavor and heating odor was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Reference example 1
  Mix 84g dried rice cake (78g dry matter weight), 50g salt, 25ml ethanol, 375ml water, adjust pH to 8.5, then add 0.3ml Alcalase 2.4L (Novo Nordisk Bio Industry Co., Ltd.) And left at 30 ° C. The liquid mixture during standing was stirred once a day. Further, the pH of the mixed solution was sometimes adjusted to 8.5 using a NaOH solution (40 w / v%). One week later, the pH of the mixed solution was adjusted to 6.0, and then 0.8 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bioindustry Co., Ltd.] was added and left to stand for 1 week to complete the reaction. . Next, after heating at 80 degreeC for 30 minutes and deactivating an enzyme, it filtered and obtained the seasoning. General analysis values are TN 1.6%, FN 0.45%, NaCl 10.0%, EtOH 5%, pH 5.8, Brix 28.1, OD₄₃₀  The seasoning excellent in flavor, umami, and kokumi without the soy sauce flavor and heating odor of 3.1 was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Example2
  150 g of bonito extract residue (TN 6%) (dry matter weight 75 g), 34 g of sodium chloride, 17 ml of ethanol and 170 ml of water were mixed, pH was adjusted to 8.5, and then Alcalase 2.4 L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] 0.3 ml was added and allowed to stand at 30 ° C. The liquid mixture during standing was stirred once a day. Further, the pH of the mixed solution was sometimes adjusted to 8.5 using a NaOH solution (40 w / v%). After 4 weeks, the pH of the mixed solution was adjusted to 5.5, and then 0.8 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bioindustry Co., Ltd.] was added and left to stand for 1 week to complete the reaction. . Next, after heating at 80 degreeC for 30 minutes and deactivating an enzyme, it filtered and obtained the seasoning. General analysis values are TN 1.9%, FN 0.8%, NaCl 10.0%, EtOH 5%, pH 5.4, Brix 25.4, OD₄₃₀  A seasoning excellent in flavor, umami, and kokumi without 5.9 soy sauce flavor and heated odor was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Reference example 2
  After mixing 86 g of corn gluten meal (TN 11.9%), 50 g of sodium chloride, 25 ml of ethanol and 373 ml of water and adjusting the pH to 9.0, Alcalase 2.4 L [manufactured by Novo Nordisk Bioindustry Co., Ltd.] 0.33 ml was added and allowed to stand at 30 ° C. The liquid mixture during standing was stirred once a day. In addition, the pH of the mixed solution was sometimes adjusted to 9.0 using a NaOH solution (40 w / v%). After 4 weeks, the pH of the mixed solution was adjusted to 5.0, and then 0.82 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bioindustry Co., Ltd.] was added and allowed to stand for 2 weeks to complete the reaction. . Next, after heating at 80 degreeC for 30 minutes and deactivating an enzyme, it filtered and obtained the seasoning. General analysis values are TN 1.7%, FN 0.51%, NaCl 10.5%, EtOH 4.5%, Brix 24.0, OD₄₃₀  A seasoning excellent in flavor, umami and kokumi without the soy sauce flavor and heating odor of 4.5 was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Example3
  181.2 g of bonito extract residue (TN 6.9%) (dry matter weight 87.0 g), 30 g of sodium chloride, 25 ml of ethanol and 282.9 ml of water were mixed, pH was adjusted to 10.0, and then 2.4 L [Novonor] Disc Bio Industry Co., Ltd.] was added in an amount of 0.33 ml, and a total of 500 ml was allowed to stand at 30 ° C. The liquid mixture during standing was stirred once a day. Further, the pH of the mixed solution was adjusted to 10.0 with a NaOH solution (40 w / v%) from time to time. After 4 weeks, the pH of the mixed solution was adjusted to 5.5, and then 0.8 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bioindustry Co., Ltd.] was added and allowed to stand for 2 weeks to complete the reaction. . Next, after heating at 80 degreeC for 30 minutes and deactivating an enzyme, it filtered and obtained the seasoning. General analysis values are TN 1.6%, FN 0.48%, NaCl 6.0%, EtOH 5%, pH 5.4, Brix 20.5, OD₄₃₀  An enzymatic degradation product excellent in flavor, umami and kokumi without 3.8 soy sauce flavor and heating odor was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Example4
  181.2 g of bonito extract residue (TN 6.9%) (dry matter weight 87.0 g), 25 g of sodium chloride, 30 ml of ethanol and 287.9 ml of water were mixed, pH was adjusted to 9.6, and then 2.4 L [Novonor] Disc Bio Industry Co., Ltd.] was added in an amount of 0.33 ml, and a total of 500 ml was allowed to stand at 30 ° C. The liquid mixture during standing was stirred once a day. Further, the pH of the mixed solution was sometimes adjusted to 9.6 using a NaOH solution (40 w / v%). After 5 weeks, the pH of the mixed solution was adjusted to 5.5, and then 0.8 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] was added and the mixture was allowed to stand for 1 week. Next, the enzyme was inactivated by heating at 80 ° C. for 30 minutes, and then filtered to obtain a seasoning. General analysis values are TN 1.7%, FN 0.49%, NaCl 5.1%, EtOH 5.9%, pH 5.3, Brix 19.6, OD.₄₃₀  An enzymatic degradation product excellent in flavor, umami and kokumi without 3.9 soy sauce flavor and heating odor was obtained. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Example5
  After adding 135 g of dried rice cake (TN 11.9%), ethanol 40 ml, and salt 80 g to 600 ml of barley shochu distillation waste liquid (TN 0.71%, Brix 24.8), the pH was adjusted to 8.6, and then Alcalase 2. 0.4 L of 4 L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] was added, and a total of 800 ml was allowed to stand at 35 ° C. The liquid mixture during standing was stirred once a day. Further, the pH of the mixed solution was sometimes adjusted to 8.6 with NaOH solution (40 w / v%). Three weeks later, after adjusting the pH to 6.5, 0.9 ml of Flavorzyme 1000 L [manufactured by Novo Nordisk Bio Industry Co., Ltd.] was added and the mixture was allowed to stand for another week. Next, the mixture was solid-liquid separated, heated at 80 ° C. for 30 minutes to deactivate the enzyme, and then filtered to obtain a seasoning. General analysis values are TN 2.2%, FN 0.8%, NaCl 10.5%, EtOH 5.5%, pH 5.8, Brix 22.0 19.6, OD.₄₃₀  It was 5.6 with no soy sauce flavor and heating odor, and was excellent in flavor and body. Moreover, no spoilage due to various bacteria during the decomposition reaction was observed.

Industrial applicability
  According to the present invention, it is possible to provide an enzymatic degradation product with good flavor, umami, and rich taste. Further, according to the present invention, it is possible to provide an effective utilization method of by-products such as koji extraction residue, shochu distillation residue and okara. Furthermore, in the present invention, it is possible to produce an economically inexpensive enzymatic degradation product without requiring a special apparatus such as an enzymatic degradation tank.

Claims

Any of the protein-containing products derived from seafood or shochu distillation residue, under the condition that the salt concentration in the enzymatic decomposition reaction solution is 8-12 w / v% and the ethanol concentration is 3.5-6 v / v% (A) A seasoning obtained by allowing endo-type protease to act under 20 to 40 ° C. and alkaline conditions, and then (b) allowing exo-type protease to act under 20 to 40 ° C. and neutral to acidic conditions.

In the fish-derived protein-containing product or shochu distillation residue, the salt concentration in the enzymatic decomposition reaction solution is 1 w / v% or more and less than 8 w / v%, and the ethanol concentration is 3.5 to 12 v / v%. Under certain conditions, (a) an endo-type protease is allowed to act at 20-40 ° C. and pH 9.5 to above -12 conditions, and then (b) an exo-type protease is acted at 20-40 ° C. and neutral to acidic conditions. Condiment that lets you.

The seasoning according to claim 1 or 2, wherein the peptide content in the seasoning is 0.4 to 0.9.

A food, beverage or seasoning comprising the seasoning according to any one of claims 1 to 3.

While maintaining the salt concentration and ethanol concentration in the enzymatic decomposition reaction solution at 8-12 w / v% and 3.5-6 v / v%, respectively, the protein content derived from fish and shellfish or the shochu distillation residue at 20-40 ° C. A method for producing a seasoning comprising the steps of: (a) causing endo-type protease to act at pH 8.5 to 9.5; and (b) allowing exo-type protease to act under neutral to acidic conditions.

A protein-containing product derived from fish and shellfish at 20 to 40 ° C. while maintaining the salt concentration and ethanol concentration in the enzymatic decomposition reaction solution at 1 w / v% or more and less than 8 w / v% and 3.5 to 12 v / v%, respectively A method for producing a seasoning comprising: (a) a step of causing an endo-type protease to act at a pH of more than 9.5 to 12 on a shochu distillation residue; and (b) a step of causing an exo-type protease to act under neutral to acidic conditions.

The method according to claim 5 or 6, wherein the peptide content in the seasoning is 0.4 to 0.9.