JP4336930B2 - Method for producing lactic acid bacteria fermentation broth and food made from the fermentation broth - Google Patents
Method for producing lactic acid bacteria fermentation broth and food made from the fermentation broth Download PDFInfo
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- JP4336930B2 JP4336930B2 JP2001190333A JP2001190333A JP4336930B2 JP 4336930 B2 JP4336930 B2 JP 4336930B2 JP 2001190333 A JP2001190333 A JP 2001190333A JP 2001190333 A JP2001190333 A JP 2001190333A JP 4336930 B2 JP4336930 B2 JP 4336930B2
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- Prior art keywords
- lactic acid
- acid bacteria
- yeast
- fermentation
- dry yeast
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Description
【0001】
【発明の属する技術分野】
本発明は、新規な乳酸菌発酵液の製造方法及び該発酵液を原料とする食品・医薬に関する。
【0002】
【従来の技術】
酵母は古くから発酵食品の製造過程で利用されており、パンやビールはもちろん、日本人の食卓には欠かせない味噌,醤油,日本酒など、酵母を利用した醸造産業は近代以前の産業界では重要な位置を占めていた。
【0003】
近年、酵母細胞内の成分の生化学的解明が進むにつれ、酵母のもつ栄養学的特性,薬理効果を期待して積極的に利用しようとする動きが盛んになってきた。酵母のもつ栄養学的特性は、(1)良質な高蛋白、必須アミノ酸をバランスよく含む、(2)豊富なビタミン,ミネラル含量、(3)核酸、多糖類に富む、などがあげられ、以上の効用から総合ビタミン補給剤,胃腸薬剤,飼料の栄養強化剤として利用されている。
【0004】
しかし、酵母を栄養強化の目的として加工食品に配合しようとした場合、酵母独特の好ましくない臭いや苦み,呈味があるため、その用途や使用量等に制約を受けることが多い。即ち、錠剤型とすると、酵母の臭い、苦みなどをあまり感じなくなるが、所望の効果を得るためには多量のタブレットを摂取する必要があり、毎日継続して摂取するのが困難である。一方、飲料の形態とすると、所要量(有効量)の摂取は容易であるが、酵母の臭い、苦みなどのため非常に飲みにくいものとなる。
【0005】
そこで、酵母のもつ不快臭のマスキング方法として、従来化学物質や香料などの添加,加熱処理,酵素処理等が提案,実施されている。しかしこれらの方法は、十分な効果が得られない、操作が煩雑で経済的でない、衛生上問題がある等の欠点があり、酵母はその高い栄養学的価値にもかかわらず、健康食品素材としては十分活用されていないのが現状である。
【0006】
一方、生酵母を酵母細胞壁溶解酵素で処理した後、乳酸発酵させる技術が提案されているが、この場合、酵素処理中の微生物汚染の問題や、酵素処理によって酵母細胞中に含まれるタンパク、ペプチド等が完全に溶出し、またその一部がアミノ酸まで分解されるので、香味の点で好ましくない。
【0007】
また、穀類,野菜,畜肉等を酵母菌,乳酸菌,麹菌等により発酵し、素材の不快味および不快臭を改良,低減する方法はこれまで種々提案されてきたが、酵母そのものの不快味,不快臭,苦みを乳酸菌発酵により改良,低減するという発想はこれまでにはなかった。
【0008】
【発明が解決しようとする課題】
本発明は、上記したような欠点のない酵母の乳酸菌発酵液の製造方法及び該発酵液を原料とする食品や医薬を提供することを目的とする。
【0009】
【課題を解決するための手段】
このような事情を鑑み、本発明者は栄養学的価値の高い酵母を手軽においしく摂取できる、望ましい加工食品を提供するために鋭意検討した結果、乾燥酵母を乳酸菌で発酵させることにより、酵母本来のもつ独特の不快臭,苦み,呈味がマスキングされた嗜好性の高い飲料が得られることを見出した。
【0010】
即ち、本発明は下記の各項に示された発明に関する。
項1 乾燥酵母を乳酸菌で発酵することを特徴とする、乳酸菌発酵液の製造方法。
項2 発酵を、好気的条件で行うことを特徴とする項1記載の製造方法。
項3 発酵を、麦芽エキスの存在下で行うことを特徴する項1または2記載の製造方法。
項4 乳酸菌としてLactobacillus rhamnosus 及びLactobacillus plantarumからなる群より選ばれる少なくとも1種を用いることを特徴とする項1〜3のいずれかに記載の製造方法。
項5 乾燥酵母が、乾燥ビール酵母であることを特徴とする項1〜4のいずれかに記載の製造方法。
項6 乾燥酵母を乳酸菌で発酵して得ることができる乳酸菌発酵液を原料とする食品。
項7 整腸作用を有する、項6に記載の食品。
項8 乾燥酵母を乳酸菌で発酵して得ることができる乳酸菌発酵液を原料として含む便秘薬。
項9 乾燥酵母を乳酸菌で発酵して得ることができる乳酸菌発酵液を原料として含む整腸薬。
【0011】
【発明の実施の形態】
本発明においては、乾燥酵母を用いる。乾燥酵母は、生酵母をドラム乾燥,噴霧乾燥,通気乾燥などの方法により乾燥,粉末化したものであり、水分が10%以下のものである。本発明では、活性乾燥酵母を用いてもよく、不活性乾燥酵母を用いてもよい。
【0012】
通常商業的に入手可能な酵母原料としては、生酵母,酵母エキス,乾燥酵母等があるが、本発明において乾燥酵母を用いる理由は以下の通りである。
【0013】
生酵母にはマンナン,グルカン,蛋白質,脂肪等からなる極めて強固な細胞壁が存在しているために消化性が悪く、菌体内に含まれる豊富な栄養成分の利用率が低いという欠点がある。酵母エキスは、生酵母を自己消化もしくは酵素分解後水溶成分を抽出したエキスであり、酵母由来の蛋白質が酵素分解等によりペプチド,アミノ酸まで分解されており、呈味が強すぎ配合量が制限されるという難点がある。一方、乾燥酵母は、その乾燥工程において強固な細胞壁の構造がルーズになり、細胞質の透過性が増大しているので消化吸収性もよく、呈味も酵母エキスほど強くない。かかる理由により、本発明において乾燥酵母を原料として用いる。
【0014】
乾燥酵母の原料としてはビール酵母、パン酵母、トルラ酵母、石油酵母、清酒酵母などが好適であるが、とりわけビールの醸造の際に副産物として得られるビール酵母は安価かつ安定的に入手できるのでより好ましい。
【0015】
また、本発明では、乳酸菌であれば特に限定されず用いることができ、通性ヘテロ発酵タイプ、絶対ホモ発酵タイプ、絶対ヘテロ発酵タイプのいずれであってもよい。これらの中でも、乾燥酵母の発酵性の点からラクトバチルス属に属する菌が好ましく、ラクトバチルス属の通性ヘテロ発酵タイプの乳酸菌がより好ましい。さらには、乾燥酵母の香味のマスキング効果からみて、ラクトバチルス・プランタラム,ラクトバチルス・ラムノサス,ラクトバチルス・カゼイ,ラクトバチルス・アリメンタリウス,ラクトバチルス・コリニフォルミスなどが好適で、特にラクトバチルス・プランタラム,ラクトバチルス・ラムノサスが好ましい。これらは単独、又は任意の組み合わせで用いることができる。
【0016】
ストレプトコッカス・ラクチス,エンテロコッカス・フェシウム等の乳酸球菌も、本発明方法において乳酸菌として用いることができるが、これら球菌は、ラクトバチルス属に属する乳酸菌と組み合わせて乾燥酵母の発酵に用いることが好ましい。
【0017】
本発明方法においては、1〜30重量%程度、好ましくは5〜20重量%程度の乾燥酵母の懸濁液を調製後、乳酸菌が資化できる糖質源を、添加後の濃度が0.5〜20重量%程度、好ましくは1〜10重量%程度となるように添加することが望ましい。糖質源としては、乳酸菌が資化できるものであれば特に限定されるものではないが、例えば、グルコース、キシロース、フラクトース、サッカロース、マルトース、トレハロース、パラチノース、ラクトース、ガラクトース、ガラクトオリゴ糖、フラクトオリゴ糖、大豆オリゴ糖、マルトオリゴ糖、イソマルトオリゴ糖、キシロオリゴ糖やこれら糖を水添した糖アルコール、麦芽エキス等が挙げられる。これらの中でも、乾燥酵母の香味のマスキング効果からみて麦芽エキスが最も好ましい。ここで、麦芽エキスは、麦芽をビール醸造における仕込み作業とほぼ同様な方法で糖化させ、糖化液を濾過後、濃縮したもの、またはさらに乾燥粉末化しものであり、食品の分野で通常用いられているものを利用することができる。これら糖質源は、単独、もしくは組み合わせて添加してよい。
【0018】
また、乾燥酵母の懸濁液の調製に際しては、栄養強化もしくは香味改善を目的として、乾燥酵母の一部に代えて、獣乳、還元脱脂乳、大豆粉末、大豆分離蛋白、豆乳、ホエイ;野菜や果物の磨砕物、搾汁、搾汁濃縮物等の1種又は2種以上を、乳酸菌による発酵を阻害しない範囲で用いることができる。
【0019】
上記のように調製された培地は、発酵に先立ち90℃、5分程度の加熱殺菌を行うことが望ましい。
【0020】
発酵は、例えば、スターターを乾燥酵母懸濁液に対し0.5〜2重量%程度接種し、定法に従って25〜42℃程度、好ましくは30〜38℃程度で行うことができる。スターターは、発酵に用いる乳酸菌を、酵母エキス、ペプトン等を添加した10〜15%還元脱脂乳等の公知の培地を用いて定法に従って培養して調製することができる。
【0021】
乳酸菌は通性嫌気性菌であり、通常の乳酸菌発酵液製造過程は嫌気条件である静置発酵で行われる。本発明の方法においても、嫌気条件である静置発酵において、乾燥酵母のマスキング効果が得られる。しかしながら、本発明者は発酵中に発酵液をプロペラ攪拌器等で攪拌するなど好気条件で発酵することにより、静置発酵に比べて一層乾燥酵母の不快臭のマスキング効果が増強されることを見出した。従って、本発明の方法において、乳酸菌を用いた発酵工程は、好気的に行うことが好ましい。
【0022】
発酵時間は乾燥酵母の濃度、その他配合成分により適宜設定できるが、通常、8〜48時間程度で、生成する乳酸量を目安に、所望の最終製品に適した発酵液となるように、発酵の終点を適宜定めればよい。
【0023】
かくして得られた乳酸菌発酵液には、酵母の味や酵母の臭みがマスキングされ、また、ビール酵母に含まれていたタンパクがアミノ酸まで分解されないのでアミノ酸の味があまりなく、香味の点で優れており、食品用の原料として好適である。
【0024】
本発明においては上記のようにして製造した乳酸菌発酵液を最終製品中の乾燥酵母含量が0.5〜10.0重量%程度になるように希釈し、また必要に応じて通常食品に用いられる甘味剤、果汁、野菜汁、酸味剤、香味剤、着色剤、強化剤等の添加剤を加えて味を整えた乳酸菌飲料を得ることができる。もちろん、発酵液をそのまま乳酸発酵飲料としてもよい。
【0025】
乳酸菌飲料は、乳を原料として用いない非乳製品の乳酸菌飲料を主とするが、乳を原料の一部として用いた乳酸菌飲料(生菌乳製品乳酸菌飲料、殺菌乳製品乳酸菌飲料など)であってもよい。
【0026】
その他の食品の形態については、本発明に係る乳酸菌発酵液を、そのまま、或いは、濃縮,乾燥させるなどして加工した後、任意の食品に配合することもできる。例えば、粉末飲料、タブレット、カプセル、キャンディー、ゼリー、ヨーグルト、チョコレート、揚げ菓子、焼き菓子、パン、ケーキ類などの形状の食品に調製することができる。これら食品においても、酵母の味や酵母の臭みがマスキングされ、また、香味の点で優れており、摂取しやすい。
【0027】
本明細書における「食品」には、乳酸菌飲料、その他上記したような乳酸菌発酵液を原料として調製された食品が含まれる。
【0028】
本発明方法により得られた乳酸菌発酵液を原料として調製された食品は、良質な蛋白、必須アミノ酸をバランスよく含み、また、ビタミン、ミネラルなどを豊富に含み、ビタミン補給、栄養強化などの作用が期待できる。
【0029】
従って、本発明の食品は、食事が偏りがちな人や食生活が不規則な人を対象とした健康食品、機能性食品、特定保健用食品又は病者用食品という用途に用いることができる。
【0030】
また、本発明方法では乾燥酵母をそのまま発酵しているため、細胞壁又はその一部が含まれる。細胞壁の主成分はβ―グルカンであり、体内で食物繊維のような働きをすることが知られているので、乳酸菌と相まって、整腸作用が発揮される。
【0031】
具体的には、下記の実施例に示すように、本発明方法により得られた乳酸菌発酵液を原料として調製された食品は、排便回数、便の形状・硬さ(柔らかさ)、便の容量、排便後の感覚(すっきり感)などの、便通・便性を改善する効果を有している。さらに、便通・便性を改善することにより、それに伴う肌荒れ、吹き出物、腹部膨満感などを改善する作用も期待できる。
【0032】
従って、本発明の食品は、整腸のための、健康食品、機能性食品、特定保健用食品又は病者用食品という用途に用いることができる。
【0033】
また、本発明方法により得られた乳酸菌発酵液を原料として調製された食品は、体内で食物繊維のような働きをするβ―グルカンを主成分とする細胞壁又はその一部が含まれているので、乳酸菌と相まって、免疫賦活作用や、免疫機能を整える作用が期待できる。
【0034】
従って、本発明の食品は、免疫強化のため、或いは免疫機能を整えるための、健康食品、機能性食品、特定保健用食品又は病者用食品という用途に用いることができる。
【0035】
乳酸菌発酵液は、食品原料以外にも、医薬品原料、培地原料、動物薬の原料、飼料原料などの用途にも用いることができる。
【0036】
例えば、上記乳酸菌飲料を原料とする医薬は、整腸したり、便通・便性を改善する作用を有しているので、便秘薬、整腸薬、下剤(瀉下薬)として有用である。
【0037】
かかる医薬組成物は、本発明の乳酸菌発酵液を医薬品原料として、これを適当な無毒性の製剤学的に許容される担体と共に用いて、一般的な医薬製剤の形態として実用される。
【0038】
上記医薬製剤の投与単位形態としては、各種の形態が選択でき、その代表的なものとしては、錠剤、丸剤、顆粒剤、カプセル剤等の固形製剤;内用散剤等の粉体製剤;液剤、懸濁剤、乳剤、シロップ剤等の液状製剤が挙げられる。
【0039】
本発明医薬製剤に利用される上記製剤学的に許容される担体としては、製剤の使用形態に応じて通常使用される、結合剤、崩壊剤、界面活性剤、吸収促進剤、吸着剤、滑沢剤、充填剤、増量剤、付湿剤等の希釈剤又は賦形剤を例示できる。これらは得られる製剤の投与単位形態に応じて適宜選択して使用される。
【0040】
更に、本発明医薬製剤には、必要に応じて防腐剤、甘味剤、着色剤、抗酸化剤、保存剤、香料、風味剤等や他の医薬品を含有させ、医薬製剤として調製することもできる。
【0041】
上記医薬製剤は、当該形態における常法に従って調製することができる。また、上記医薬製剤は、通常経口投与される。
【0042】
本発明の食品又は医薬製剤の摂取量については、摂取の目的、食品又は医薬製剤の形態、摂取者の年齢、性別その他の条件に応じて適宜選択されるが、通常、乾燥酵母量として、1日成人1人当たり0.01〜20g程度、好ましくは0.1〜10g程度となるような量とするのがよく、1日に1回又は2〜4回に分けて摂取することができる。
【0043】
本発明食品又は医薬製剤は、その目的に応じて摂取期間を設定することができる。
【0044】
例えば、便秘の人が便秘の症状を改善するためには、症状が改善されるまで1日〜12週間程度、好ましくは3日〜3週間程度連続的に摂取することが好ましく、改善後も、1週間に1〜7日、好ましくは2〜4日程度摂取して排便習慣を維持するのが望ましい。
【0045】
便秘ぎみの人、便秘がちな人、便秘になりやすい人、便秘傾向にある人などは、排便習慣を維持するために、1週間に1〜7日程度、好ましくは2〜4日程度の割合で、本発明の食品又は医薬製剤を継続的に摂取して、お腹の調子を整えたり便秘になりにくいようにすることができる。
【0046】
また、軟便の人が便の硬さを改善するためには、便の硬さが改善されるまで、通常1日〜24週間程度、好ましくは3日〜12週間程度連続的に摂取することが好ましく、改善後も、1週間に1〜7日程度、好ましくは2〜4日程度摂取して排便習慣を維持するのが望ましい。
【0047】
軟便ぎみの人、軟便がちな人、軟便になりやすい人、軟便傾向にある人などは、軟便にならないように、1週間に1〜7日、好ましくは2〜4日程度の割合で、本発明の食品又は医薬製剤を継続的に摂取することができる。
【0048】
【実施例】
以下、実施例及び比較例を挙げて本発明の効果をさらに詳しく説明するが、本発明はこれらに限定されるものではない。なお、「%」は特に断らない限り重量%を意味する。
【0049】
試験例1
種々の乳酸菌を用いて乾燥酵母の発酵試験を行った。
【0050】
乾燥ビール酵母15%,グルコースを3%添加した懸濁液を調整後、90℃,10分の殺菌を行い、すみやかに冷却して乳酸菌発酵培地とした。これらの培地に対して、あらかじめ培養しておいた9種の乳酸菌スターターをそれぞれ1%接種し、37℃,28時間静置発酵した。スターターは、発酵に用いる乳酸菌を、酵母エキスを添加した12%還元脱脂乳からなる培地を用いて、36℃で20時間培養して調製した。
【0051】
発酵終了後の発酵液の乳酸酸度、および香味評価結果を以下の表1に示す。なお香味評価は専門パネラー6名の、1(好ましくない)から5(好ましい)の5段階評価の合計点でスコア化した。
【0052】
【表1】
【0053】
未発酵の比較例1は、酵母の味が強かったが、乳酸菌発酵させた実施例1〜9は、比較例1と比べて酵母の味がほとんどなく、酵母の臭い及び味のマスキング効果が示された。また、表1に示すごとく、ラクトバチルス属の通性ヘテロ発酵タイプの菌(コリニフォルミス,アリメンタリウス,カゼイ,ラムノサス及びプランタラム)良好な酵母臭乃至酵母味のマスキング効果が認められた。
【0054】
試験例2
発酵時の発酵糖質源の種類及び発酵条件について検討を行った。
【0055】
試験例1の発酵糖質源を種々の糖質(固形分換算で3%添加)に置き換えた培地を同様に調整し、ラクトバチルス・ラムノサスをスターターとして37℃,20時間、静置発酵(実施例10〜17),またはスターラーにて攪拌条件下(実施例18〜25)で発酵を行った。発酵終了後の発酵液の乳酸酸度、および香味評価結果を以下に示す。なお香味評価はさらに詳細に解析を行うべく、5種の香味の項目(エグ味,苦味,ざらつき,呈味,生臭さ)について、専門パネラー6名の1(好ましくない)から5(好ましい)の5段階評価の合計点でスコア化した。結果を表2及び表3に示す。
【0056】
【表2】
【0057】
【表3】
【0058】
表2,表3に示すごとく、全般的に良好な香味の発酵液が得られたが、発酵糖質源に麦芽エキスを用いること、および攪拌発酵を行うことによりさらに高い香味改善が確認された。
【0059】
実施例26
実施例25で得た乾燥酵母の乳酸菌発酵液を用い、下記の処方により各成分を混合して、常法により乳酸菌飲料を作成した。
【0060】
試験例3
実施例26で得た飲料A、飲料Aの乳酸菌発酵液を実施例17で得られた発酵液に置き換えた飲料B(実施例27)、実施例10で得られた発酵液に置き換えた飲料C(実施例28)、比較例2の乳酸菌未発酵液に置き換えた飲料C(比較例3)を用い、一般消費者50名(男性25名,女性25名:平均年齢36歳)を対象として官能評価を行った。結果を表4に示す。
【0061】
【表4】
【0062】
表4の結果から判るように、通常香味に劣る乾燥酵母を、乳酸菌、特にラクトバチルス属の通性ヘテロタイプの乳酸菌で発酵することにより、酵母のもつ不快臭,不快味をマスキングし、飛躍的に嗜好性に優れた酵母含有飲料を得ることができ、さらには発酵等質源に麦芽エキスを用いる,または好気(攪拌)条件にて発酵することによりそのマスキング効果を向上できることが明らかである。
【0063】
試験例4
便秘症状を自覚する成人を対象として、乾燥ビール酵母錠剤及び乾燥ビール酵母の乳酸菌発酵液を原料とする飲料による、排便(便通・便性)に及ぼす影響について検討した。
【0064】
乾燥ビール酵母錠剤としては、乾燥ビール酵母を錠剤の製造に慣用されている担体を用いて錠剤の形態として得られたものを用いた。乳酸菌発酵ビール酵母飲料としては、前記乾燥ビール酵母を乳酸菌(Lactobacillus rhamnosus)で発酵して得られた乳酸菌発酵液を配合して加熱殺菌して得られた飲料を用いた。
【0065】
9名のボランティアに乾燥ビール酵母錠剤を1日25錠(乾燥酵母量として3.75g)<以下、“錠剤群”とする>、8名に乳酸菌発酵ビール酵母飲料を1日1缶(160g、乾燥酵母量として3.75g)<以下、“飲料群”とする>、それぞれ3週間摂取させた。錠剤群には、摂取時に飲料とほぼ同量の水も摂取させた。摂取期間の前に2週間の非摂取期間を設け、排便に関するアンケート調査を実施した。排便量はφ25mm×85mmの円柱本数に見たてて記入させた(1:1本未満、2:2本未満、3:3本未満、4:4本未満、5:4本以上)。排便後の感覚は、1:すっきり、2:普通、3:すっきりしない、という基準により記入させた。試験期間中、乳酸菌製剤、乳酸菌飲料、食物繊維、オリゴ糖配合飲料、糖アルコール強化飲料・食品、野菜ジュース、医薬品(抗生物質、整腸剤、下剤)など便通・便性に影響を与える可能性がある食品や薬剤の摂取を控えさせた以外は特に食事制限を行わなかった。摂取前2週間のうち1週目は調整期間とし、摂取前2週目を対照期間として設定した。
【0066】
各群について統計学的検定を行った結果、飲料群について、摂取期間中に排便回数、容量、排便後の感覚(すっきり感)に有意な改善が認められた。錠剤群に関しては、摂取により便通改善の傾向は見られたが、有意な差ではなかった。また、飲み易さに関しては、錠剤群は錠剤を25錠毎日飲むのが困難であるボランティアもいたが、飲料群は1日摂取量が160gであり、臭みや苦みなどが抑えられているので毎日飲用するのは特に大変ではなかった。
【0067】
試験例5
軟便の成人を対象として、試験例4と同一の,乾燥ビール酵母の乳酸菌発酵液を原料とする飲料が、便性に及ぼす影響について検討した。
【0068】
7名のボランティアに乳酸菌発酵ビール酵母飲料を1日1缶(160g、乾燥酵母量として3.75g)を3ヶ月間摂取させた。摂取期間の前に2週間非摂取期間を設け、便性に関するアンケート調査を実施した。便の形状及び硬さについて下記の評価に従って記録させた。
便の形状:1.水状 2.泥状 3.半練り状 4.バナナ状 5.カチカチ状
6.コロコロ状
便の硬さ:1.非常に柔らかい 2.軟らかい 3.ふつう 4.硬い 5.非常に硬い
結果を表5に示す。
【0069】
【表5】
【0070】
統計学的検定を行った結果、摂取期間中に便の形状及び硬さについて、保形性が改善される方向で有意差が認められた。
【0071】
以上の結果より、本発明の乳酸菌発酵ビール酵母飲料は、便通乃至便性の改善に有用であることが明らかになった。
【0072】
【発明の効果】
本発明によれば、栄養学的価値が高いが加工食品として摂取し難い乾燥酵母を、乳酸菌発酵することにより、嗜好性に優れた乳酸菌飲料などの乳酸菌発酵液を原料とする食品や医薬製剤が提供される。また、本発明の乳酸菌発酵液を原料とする食品や医薬製剤によれば、整腸作用が得られ、便通・便性が改善する。
【図面の簡単な説明】
【図1】乳酸菌発酵ビール酵母飲料による排便回数の変化を示す図。
【図2】酵母錠剤摂取による排便回数の変化を示す図。
【図3】乳酸菌発酵ビール酵母飲料による排便容量の変化を示す図。
【図4】酵母錠剤摂取による排便容量の変化を示す図。
【図5】乳酸菌発酵ビール酵母飲料による排便後の感覚(すっきり感)の変化を示す図。
【図6】酵母錠剤摂取による排便後の感覚(すっきり感)の変化を示す図。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a novel method for producing a lactic acid bacteria fermentation broth and food / medicine using the fermentation broth as a raw material.
[0002]
[Prior art]
Yeast has long been used in the manufacturing process of fermented foods, and brewing industries using yeast, such as miso, soy sauce, and sake, which are indispensable for Japanese tables, as well as bread and beer, are pre-modern industries. Occupied an important position.
[0003]
In recent years, with the progress of biochemical elucidation of components in yeast cells, the movement to actively use them in anticipation of the nutritional characteristics and pharmacological effects of yeast has become active. The nutritional characteristics of yeast include (1) high-quality high protein, well-balanced essential amino acids, (2) rich vitamin and mineral content, (3) rich in nucleic acids and polysaccharides, etc. It is used as a general vitamin supplement, gastrointestinal drug, and feed fortification because of its benefits.
[0004]
However, when trying to add yeast to processed foods for the purpose of enhancing nutrition, there are many unfavorable odors, bitterness, and tastes unique to yeast, and there are many restrictions on their use and usage. That is, when it is a tablet type, it does not feel the smell or bitterness of yeast so much, but in order to obtain a desired effect, it is necessary to ingest a large amount of tablets, and it is difficult to ingest continuously every day. On the other hand, in the form of a beverage, the required amount (effective amount) can be easily taken, but it is very difficult to drink due to the smell of the yeast, bitterness and the like.
[0005]
Therefore, as a masking method for unpleasant odors possessed by yeast, the addition of chemical substances and fragrances, heat treatment, enzyme treatment, and the like have been proposed and implemented. However, these methods have drawbacks such as insufficient effects, complicated operation and economicalness, and sanitary problems. Yeast is a health food material despite its high nutritional value. Is not fully utilized.
[0006]
On the other hand, a technique has been proposed in which live yeast is treated with a yeast cell wall lytic enzyme and then lactic acid fermented. In this case, the problem of microbial contamination during the enzyme treatment and the proteins and peptides contained in the yeast cell by the enzyme treatment are proposed. Etc. are completely eluted, and some of them are degraded to amino acids, which is not preferable in terms of flavor.
[0007]
In addition, various methods for improving and reducing unpleasant taste and unpleasant odor of materials by fermenting cereals, vegetables, livestock meat, etc. with yeast, lactic acid bacteria, and koji molds have been proposed. There has never been an idea to improve or reduce odor and bitterness by fermentation of lactic acid bacteria.
[0008]
[Problems to be solved by the invention]
An object of this invention is to provide the manufacturing method of the lactic-acid-bacteria fermented liquor of yeast without the above faults, and the foodstuffs and medicine which use this fermented liquid as a raw material.
[0009]
[Means for Solving the Problems]
In view of such circumstances, the present inventor has intensively studied to provide a desirable processed food that can be easily and deliciously ingested with high nutritional value yeast. It has been found that a beverage with high palatability can be obtained in which the peculiar unpleasant odor, bitterness and taste are masked.
[0010]
That is, this invention relates to the invention shown by each following item.
Item 4. The method according to any one of
Item 6 A food using as a raw material a fermentation liquid of lactic acid bacteria that can be obtained by fermenting dry yeast with lactic acid bacteria.
[0011]
DETAILED DESCRIPTION OF THE INVENTION
In the present invention, dry yeast is used. Dry yeast is obtained by drying and pulverizing live yeast by methods such as drum drying, spray drying, and aeration drying, and has a moisture content of 10% or less. In the present invention, active dry yeast may be used, or inactive dry yeast may be used.
[0012]
Usually, commercially available yeast raw materials include live yeast, yeast extract, and dry yeast. The reason for using dry yeast in the present invention is as follows.
[0013]
Live yeast has the disadvantage that it has poor digestibility due to the presence of extremely strong cell walls composed of mannan, glucan, protein, fat, etc., and the utilization rate of abundant nutrients contained in the cells is low. Yeast extract is an extract obtained by extracting water-soluble components after self-digestion or enzymatic decomposition of live yeast. Proteins derived from yeast are decomposed to peptides and amino acids by enzymatic decomposition, etc. There is a difficulty that. On the other hand, dry yeast has a strong cell wall structure in the drying step and looses cytoplasmic permeability, so that it has good digestibility and taste and is not as strong as yeast extract. For this reason, dry yeast is used as a raw material in the present invention.
[0014]
Beer yeast, baker's yeast, torula yeast, petroleum yeast, sake yeast, etc. are suitable as raw materials for dry yeast, but especially beer yeast obtained as a by-product during brewing beer can be obtained cheaply and stably. preferable.
[0015]
Moreover, in this invention, if it is a lactic acid bacterium, it can be used without limitation, and any of facultative heterofermentation type, absolute homofermentation type, and absolute heterofermentation type may be sufficient. Among these, bacteria belonging to the genus Lactobacillus are preferable from the viewpoint of fermentability of dry yeast, and facultative heterofermentative lactic acid bacteria belonging to the genus Lactobacillus are more preferable. Furthermore, from the viewpoint of the masking effect of the flavor of dry yeast, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus amentalius, Lactobacillus coriniformis are preferred, especially Lactobacillus -Plantaram, Lactobacillus rhamnosus are preferred. These can be used alone or in any combination.
[0016]
Lactococci such as Streptococcus lactis and Enterococcus faecium can also be used as lactic acid bacteria in the method of the present invention, but these cocci are preferably used in combination with lactic acid bacteria belonging to the genus Lactobacillus for fermentation of dry yeast.
[0017]
In the method of the present invention, after preparing a dry yeast suspension of about 1 to 30% by weight, preferably about 5 to 20% by weight, a saccharide source capable of assimilating lactic acid bacteria is added at a concentration after addition of 0.5. It is desirable to add so as to be about -20% by weight, preferably about 1-10% by weight. The carbohydrate source is not particularly limited as long as it can assimilate lactic acid bacteria.For example, glucose, xylose, fructose, saccharose, maltose, trehalose, palatinose, lactose, galactose, galactooligosaccharide, fructooligosaccharide, Examples include soybean oligosaccharides, maltooligosaccharides, isomaltooligosaccharides, xylo-oligosaccharides, sugar alcohols obtained by hydrogenation of these sugars, and malt extracts. Among these, malt extract is most preferable in view of the masking effect of the flavor of dry yeast. Here, the malt extract is obtained by saccharifying malt in substantially the same manner as in beer brewing, filtering the saccharified solution, concentrating it, or further pulverizing it, and is usually used in the field of food. You can use what you have. These carbohydrate sources may be added alone or in combination.
[0018]
When preparing dry yeast suspensions, animal milk, reduced skim milk, soy powder, soy protein isolate, soy milk, whey; vegetables instead of a part of dry yeast for the purpose of enhancing nutrition or improving
[0019]
The medium prepared as described above is preferably sterilized by heating at 90 ° C. for about 5 minutes prior to fermentation.
[0020]
For example, fermentation can be performed at about 25 to 42 ° C., preferably about 30 to 38 ° C. according to a conventional method by inoculating a starter with about 0.5 to 2% by weight of the dry yeast suspension. The starter can be prepared by culturing lactic acid bacteria used for fermentation according to a conventional method using a known medium such as 10-15% reduced skim milk supplemented with yeast extract, peptone and the like.
[0021]
Lactic acid bacteria are facultative anaerobic bacteria, and the normal lactic acid bacteria fermentation liquid production process is performed by stationary fermentation which is anaerobic conditions. Also in the method of this invention, the masking effect of dry yeast is acquired in stationary fermentation which is anaerobic conditions. However, the present inventor shows that the unpleasant odor masking effect of dry yeast is further enhanced by fermentation under aerobic conditions such as stirring the fermentation solution with a propeller stirrer during fermentation. I found it. Therefore, in the method of the present invention, the fermentation step using lactic acid bacteria is preferably performed aerobically.
[0022]
Fermentation time can be set as appropriate depending on the concentration of dry yeast and other ingredients, but it is usually about 8 to 48 hours, with the amount of lactic acid produced as a guideline, so that the fermentation solution is suitable for the desired final product. What is necessary is just to determine an end point suitably.
[0023]
In the lactic acid bacteria fermentation broth thus obtained, the taste and odor of yeast are masked, and the protein contained in brewer's yeast is not degraded to amino acids, so the taste of amino acids is not so great and excellent in flavor. It is suitable as a raw material for food.
[0024]
In the present invention, the lactic acid bacteria fermentation broth produced as described above is diluted so that the dry yeast content in the final product is about 0.5 to 10.0% by weight, and is usually used for food as necessary. Additives such as sweeteners, fruit juices, vegetable juices, acidulants, flavoring agents, coloring agents, and fortifying agents can be added to obtain a lactic acid bacteria beverage with a conditioned taste. Of course, the fermented liquid may be used as it is as a lactic acid fermented beverage.
[0025]
Lactic acid bacteria drinks are mainly non-dairy lactic acid bacteria drinks that do not use milk as raw materials, but are lactic acid bacteria drinks that use milk as part of the raw materials (eg, live lactic acid product lactic acid bacteria drinks and sterilized dairy product lactic acid bacteria drinks). May be.
[0026]
About the form of another foodstuff, after processing the lactic-acid-bacteria fermented liquid which concerns on this invention as it is or concentrating and drying, it can also mix | blend with arbitrary foodstuffs. For example, it can be prepared into foods in the form of powdered beverages, tablets, capsules, candy, jelly, yogurt, chocolate, fried confectionery, baked confectionery, bread, cakes and the like. Also in these foods, the taste of yeast and the smell of yeast are masked, and they are excellent in flavor and easy to ingest.
[0027]
“Food” in the present specification includes lactic acid bacteria beverages and other foods prepared using lactic acid bacteria fermentation liquid as described above as a raw material.
[0028]
Foods prepared using the lactic acid bacteria fermentation broth obtained by the method of the present invention as a raw material contain a good balance of high-quality proteins and essential amino acids, and are rich in vitamins, minerals, etc., and have effects such as vitamin supplementation and nutritional enhancement. I can expect.
[0029]
Therefore, the food of the present invention can be used in applications such as health foods, functional foods, foods for specified health use, or foods for the sick, for people who tend to have a biased diet or who have irregular eating habits.
[0030]
Moreover, in the method of this invention, since dry yeast is fermented as it is, a cell wall or its part is contained. The main component of the cell wall is β-glucan, and it is known that it functions like dietary fiber in the body.
[0031]
Specifically, as shown in the examples below, food prepared using the lactic acid bacteria fermentation liquid obtained by the method of the present invention as a raw material has the number of defecations, stool shape / hardness (softness), stool volume It also has the effect of improving bowel movements and stool properties such as feeling after stool (clean feeling). Furthermore, by improving the bowel movement and stool properties, the effect of improving the associated rough skin, pimples, abdominal fullness and the like can be expected.
[0032]
Therefore, the food of the present invention can be used for use as a health food, a functional food, a food for specified health use, or a food for sick people for intestinal regulation.
[0033]
In addition, the food prepared using the lactic acid bacteria fermentation broth obtained by the method of the present invention as a raw material contains a cell wall mainly composed of β-glucan that functions like dietary fiber in the body or a part thereof. In combination with lactic acid bacteria, an immunostimulatory effect and an effect of regulating immune function can be expected.
[0034]
Therefore, the food of the present invention can be used for health foods, functional foods, foods for specified health use, or foods for sick people for enhancing immunity or adjusting immune functions.
[0035]
In addition to food raw materials, lactic acid bacteria fermentation broth can be used for pharmaceutical raw materials, medium raw materials, animal drug raw materials, feed raw materials and the like.
[0036]
For example, a medicine made from the above lactic acid bacteria beverage is useful as a constipation medicine, an intestinal medicine, or a laxative (a laxative) because it has the effect of adjusting the bowel or improving bowel movements and feces.
[0037]
Such a pharmaceutical composition is practically used in the form of a general pharmaceutical preparation by using the lactic acid bacteria fermentation broth of the present invention as a pharmaceutical raw material together with a suitable non-toxic pharmaceutically acceptable carrier.
[0038]
As the dosage unit form of the pharmaceutical preparation, various forms can be selected. Typical examples thereof include solid preparations such as tablets, pills, granules, capsules; powder preparations such as internal powders; , Liquid preparations such as suspensions, emulsions, and syrups.
[0039]
Examples of the pharmaceutically acceptable carrier used in the pharmaceutical preparation of the present invention include a binder, a disintegrant, a surfactant, an absorption accelerator, an adsorbent, a lubricant, and the like that are usually used depending on the use form of the preparation. Illustrative of diluents or excipients such as bulking agents, fillers, bulking agents, moistening agents and the like. These are appropriately selected and used depending on the dosage unit form of the resulting preparation.
[0040]
Furthermore, the pharmaceutical preparation of the present invention may contain preservatives, sweeteners, coloring agents, antioxidants, preservatives, flavors, flavoring agents, and other pharmaceuticals as necessary, and can be prepared as pharmaceutical preparations. .
[0041]
The pharmaceutical preparation can be prepared according to a conventional method in the form. Moreover, the said pharmaceutical formulation is normally administered orally.
[0042]
The intake of the food or pharmaceutical preparation of the present invention is appropriately selected according to the purpose of intake, the form of the food or pharmaceutical preparation, the age of the intaker, sex, and other conditions. The amount may be about 0.01 to 20 g, preferably about 0.1 to 10 g per day, and may be taken once a day or divided into 2 to 4 times a day.
[0043]
The ingestion period of the food or pharmaceutical preparation of the present invention can be set according to the purpose.
[0044]
For example, in order for a person with constipation to improve symptoms of constipation, it is preferably taken continuously for about 1 day to 12 weeks, preferably about 3 days to 3 weeks until the symptoms are improved, It is desirable to maintain a defecation habit by ingesting about 1 to 7 days, preferably about 2 to 4 days per week.
[0045]
Persons with constipation, those who tend to have constipation, those who are prone to constipation, and those who tend to have constipation have a rate of about 1 to 7 days per week, preferably about 2 to 4 days, in order to maintain their bowel habits. Thus, the food or pharmaceutical preparation of the present invention can be continuously ingested to make the stomach well-tuned and less likely to cause constipation.
[0046]
Moreover, in order to improve the hardness of stool, a person with soft stool can usually take continuously for about 1 day to 24 weeks, preferably about 3 days to 12 weeks until the stool hardness is improved. Preferably, even after improvement, it is desirable to maintain the bowel habit by ingesting about 1 to 7 days per week, preferably about 2 to 4 days.
[0047]
Persons with loose stools, people who tend to have loose stools, people who tend to have loose stools, people with a tendency to loose stools, etc., at a rate of about 1 to 7 days per week, preferably about 2 to 4 days so as not to become loose stools The food or pharmaceutical preparation of the invention can be taken continuously.
[0048]
【Example】
Hereinafter, although an example and a comparative example are given and the effect of the present invention is explained in detail, the present invention is not limited to these. “%” Means% by weight unless otherwise specified.
[0049]
Test example 1
A dry yeast fermentation test was performed using various lactic acid bacteria.
[0050]
After preparing a suspension containing 15% dry brewer's yeast and 3% glucose, sterilization was performed at 90 ° C. for 10 minutes, and the mixture was immediately cooled to obtain a lactic acid bacteria fermentation medium. Each of these mediums was inoculated with 1% of each of 9 types of lactic acid bacteria starters that had been cultured in advance, and subjected to static fermentation at 37 ° C. for 28 hours. The starter was prepared by culturing lactic acid bacteria used for fermentation at 36 ° C. for 20 hours using a medium consisting of 12% reduced skim milk added with yeast extract.
[0051]
Table 1 below shows the lactic acid acidity and flavor evaluation results of the fermentation broth after the end of fermentation. In addition, flavor evaluation was scored by the total score of 5 grades of 1 (not preferable) to 5 (preferable) of 6 professional panelists.
[0052]
[Table 1]
[0053]
Although the unfermented Comparative Example 1 had a strong yeast taste, Examples 1 to 9 fermented with lactic acid bacteria had little yeast taste compared to Comparative Example 1, and showed a yeast odor and taste masking effect. It was done. In addition, as shown in Table 1, a facultative heterofermentative bacterium of the genus Lactobacillus (Coriniformis, Amententarius, Casei, Rhamnosus and Plantarum) showed a good yeast odor or yeast taste masking effect.
[0054]
Test example 2
The type of fermentation carbohydrate source and fermentation conditions during fermentation were examined.
[0055]
A medium in which the fermentation sugar source of Test Example 1 was replaced with various sugars (added 3% in terms of solid content) was similarly prepared, and the fermentation was performed at 37 ° C. for 20 hours using Lactobacillus rhamnosus as a starter. Fermentation was performed under stirring conditions (Examples 18 to 25) using Examples 10 to 17) or a stirrer. The lactic acid acidity of the fermentation broth after the end of fermentation and the flavor evaluation results are shown below. In addition, in order to analyze the flavor in more detail, the five types of flavor items (eg, taste, bitterness, graininess, taste, fresh odor) were evaluated from 1 (not preferred) to 5 (preferred) of 6 professional panelists. The scores were scored with a total score of 5 grades. The results are shown in Tables 2 and 3.
[0056]
[Table 2]
[0057]
[Table 3]
[0058]
As shown in Tables 2 and 3, fermented liquor with a generally good flavor was obtained, but a higher flavor improvement was confirmed by using malt extract as a fermented sugar source and by performing stirring fermentation. .
[0059]
Example 26
Using the lactic acid bacteria fermentation liquid of dry yeast obtained in Example 25, each component was mixed according to the following formulation, and a lactic acid bacteria beverage was prepared by a conventional method.
[0060]
Test example 3
Beverage A obtained in Example 26, Beverage B obtained by replacing the lactic acid bacteria fermentation liquid of Beverage A with the fermented liquid obtained in Example 17 and Beverage C replaced with the fermented liquid obtained in Example 10 (Example 28), using the beverage C (Comparative Example 3) replaced with the unfermented lactic acid bacteria of Comparative Example 2, and targeting 50 general consumers (25 men, 25 women: average age 36 years) Evaluation was performed. The results are shown in Table 4.
[0061]
[Table 4]
[0062]
As can be seen from the results in Table 4, by drying fermented yeast, which is usually inferior in flavor, with lactic acid bacteria, particularly facultative lactic acid bacteria of the genus Lactobacillus, masking the unpleasant odor and unpleasant taste of yeast, It is clear that a yeast-containing beverage with excellent palatability can be obtained, and that the masking effect can be improved by using a malt extract as the source of fermentation or fermenting under aerobic (stirring) conditions .
[0063]
Test example 4
For adults who are aware of constipation symptoms, we investigated the effects of dried beer yeast tablets and beverages made from lactic acid fermentation broth of dried beer yeast on defecation (fecal stool).
[0064]
As the dry brewer's yeast tablet, a product obtained by using dried brewer's yeast in the form of a tablet using a carrier commonly used for tablet production was used. As the lactic acid bacteria fermented beer yeast beverage, a beverage obtained by blending a lactic acid bacteria fermentation solution obtained by fermenting the dried beer yeast with lactic acid bacteria ( Lactobacillus rhamnosus ) and heat sterilizing it was used.
[0065]
Nine volunteers with 25 dry brewer's yeast tablets a day (3.75 g as dry yeast amount) <hereinafter referred to as “tablet group”>, 8 with 1 lactic acid bacteria fermented beer yeast beverage per day (160 g, The amount of dry yeast was 3.75 g) <hereinafter referred to as “beverage group”>, each of which was ingested for 3 weeks. The tablet group also received almost the same amount of water as the drink at the time of ingestion. A 2-week non-intake period was set before the intake period, and a questionnaire survey on defecation was conducted. The amount of defecation was entered by looking at the number of cylinders of φ25 mm × 85 mm (less than 1: 1, 2: less than 2, 3: less than 3, 4: less than 4, 5: 4 or more). The sensation after defecation was entered according to the following criteria: 1: clear, 2: normal, 3: unclean. During the test period, lactic acid bacteria preparations, lactic acid bacteria drinks, dietary fibers, oligosaccharide-containing drinks, sugar alcohol-enriched drinks / foods, vegetable juices, pharmaceuticals (antibiotics, intestinal preparations, laxatives), etc. There was no particular dietary restriction except withholding food and drugs. Of the 2 weeks before intake, the first week was set as the adjustment period, and the second week before intake was set as the control period.
[0066]
As a result of performing a statistical test for each group, significant improvements were observed in the number of defecations, volume, and feeling after stool (clean feeling) during the intake period for the drink group. Regarding the tablet group, there was a tendency to improve bowel movement by ingestion, but this was not a significant difference. Regarding the ease of drinking, there were volunteers who were difficult to take 25 tablets daily in the tablet group, but the daily intake of the drink group was 160 g, and the smell and bitterness were suppressed every day. It was not particularly difficult to drink.
[0067]
Test Example 5
For the soft stool adults, the same effect as in Test Example 4 on the effect of the beverage made from the lactic acid bacteria fermentation liquid of dry brewer's yeast on the stool properties was examined.
[0068]
Seven volunteers ingested 1 can (160 g, 3.75 g of dry yeast amount) of lactic acid bacteria fermented beer yeast beverage for 3 months a day. A 2-week non-intake period was set before the intake period, and a questionnaire survey on stool properties was conducted. The shape and hardness of the stool was recorded according to the following evaluation.
Stool shape: 1.
[0069]
[Table 5]
[0070]
As a result of the statistical test, a significant difference was observed in the direction in which the shape retention was improved with respect to the shape and hardness of the stool during the intake period.
[0071]
From the above results, it was revealed that the lactic acid bacteria-fermented beer yeast beverage of the present invention is useful for improving bowel movement or stool property.
[0072]
【The invention's effect】
According to the present invention, there is provided a food or pharmaceutical preparation using a fermented lactic acid bacterium such as a lactic acid bacterium beverage having excellent palatability by fermenting lactic acid bacteria with dry yeast that has high nutritional value but is difficult to ingest as processed food. Provided. Moreover, according to the foodstuff and pharmaceutical formulation which use the lactic-acid-bacteria fermented liquor of this invention as a raw material, an intestinal regulation effect is acquired and a bowel movement and feces are improved.
[Brief description of the drawings]
FIG. 1 is a graph showing changes in the number of defecations caused by lactic acid bacteria-fermented beer yeast beverages.
FIG. 2 is a graph showing changes in the number of defecations due to ingestion of yeast tablets.
FIG. 3 is a graph showing changes in defecation capacity due to lactic acid bacteria-fermented beer yeast beverages.
FIG. 4 is a graph showing changes in defecation volume due to ingestion of yeast tablets.
FIG. 5 is a view showing a change in feeling (clean feeling) after defecation by a lactic acid bacteria fermented beer yeast beverage.
FIG. 6 is a diagram showing a change in feeling (clean feeling) after defecation due to ingestion of yeast tablets.
Claims (10)
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| JP4301545B2 (en) * | 2003-01-10 | 2009-07-22 | 三井製糖株式会社 | How to improve the flavor of fermented soymilk |
| KR100649855B1 (en) * | 2003-03-07 | 2006-11-28 | 가부시키가이샤 아우레오 | Composition containing ?-glucan and constipation-relieving drug, immunopotentiator and skin moistening agent using the composition |
| JP4000078B2 (en) * | 2003-03-07 | 2007-10-31 | 株式会社アウレオ | Skin moisturizer |
| JP4054697B2 (en) * | 2003-03-07 | 2008-02-27 | 株式会社アウレオ | Constipation improving agent |
| JP5722282B2 (en) * | 2004-12-24 | 2015-05-20 | 株式会社明治 | Skin improving agent and skin improving method |
| JP5813304B2 (en) * | 2009-10-13 | 2015-11-17 | 三栄源エフ・エフ・アイ株式会社 | Method for masking unpleasant taste of collagen |
| JP5671652B2 (en) * | 2012-03-28 | 2015-02-18 | サッポロビール株式会社 | Plant raw material liquid and beverage and method related thereto |
| DE102013008335A1 (en) * | 2013-01-29 | 2014-07-31 | Erdinger Weissbräu Franz Brombach e.K. | Process for the preparation of a food as well as the food produced therewith and use of lactic acid bacteria, an acid, a mash, a wort, a drink or a concentrate |
| DE102013100891A1 (en) * | 2013-01-29 | 2014-07-31 | Erdinger Weissbräu Franz Brombach e.K. | Process for the preparation of an acid, a mash or wort, a drink, a concentrate and the products produced therewith and use of lactic acid bacteria, an acid, a mash, a wort or a concentrate |
| JP6541944B2 (en) * | 2014-06-13 | 2019-07-10 | テーブルマーク株式会社 | Deterioration inhibitor |
| KR102063248B1 (en) * | 2019-02-18 | 2020-01-07 | 이상복 | Powdered yogurt and its manufacturing method |
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