JP7444949B2 - Medium for lactic acid bacteria - Google Patents
Medium for lactic acid bacteria Download PDFInfo
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- JP7444949B2 JP7444949B2 JP2022168178A JP2022168178A JP7444949B2 JP 7444949 B2 JP7444949 B2 JP 7444949B2 JP 2022168178 A JP2022168178 A JP 2022168178A JP 2022168178 A JP2022168178 A JP 2022168178A JP 7444949 B2 JP7444949 B2 JP 7444949B2
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- lactic acid
- acid bacteria
- medium
- lactobacillus
- mass
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims description 186
- 241000894006 Bacteria Species 0.000 title claims description 122
- 235000014655 lactic acid Nutrition 0.000 title claims description 93
- 239000004310 lactic acid Substances 0.000 title claims description 93
- 239000002612 dispersion medium Substances 0.000 claims description 35
- 239000002609 medium Substances 0.000 claims description 35
- 239000012138 yeast extract Substances 0.000 claims description 22
- 235000010469 Glycine max Nutrition 0.000 claims description 20
- 229940041514 candida albicans extract Drugs 0.000 claims description 20
- 244000068988 Glycine max Species 0.000 claims description 19
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 12
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 12
- 238000004519 manufacturing process Methods 0.000 claims description 11
- 241000186660 Lactobacillus Species 0.000 claims description 10
- 239000003995 emulsifying agent Substances 0.000 claims description 10
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- 235000007340 Hordeum vulgare Nutrition 0.000 claims description 9
- 150000001720 carbohydrates Chemical class 0.000 claims description 9
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- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 7
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- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 7
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- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical group CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 2
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- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
本発明は、乳酸菌の培養に好適な乳酸菌用培地に関する。 The present invention relates to a medium for lactic acid bacteria suitable for culturing lactic acid bacteria.
これまで乳酸菌の培養に用いられる培地としては、主にMRS(deMan、Rogosa、Sharpe)培地が、すべての乳酸菌の発育を促すため、広く用いられている。 Until now, as a medium used for culturing lactic acid bacteria, MRS (deMan, Rogosa, Sharpe) medium has been widely used because it promotes the growth of all lactic acid bacteria.
このMRS培地で乳酸菌を培養し、菌数を測定したり、物性を測定するには問題はないが、MRS培地には、食品素材として使用できない試薬が含まれているため、培養した乳酸菌を直接食品用途に用いることができなかった。 There is no problem in culturing lactic acid bacteria in this MRS medium and measuring the number of bacteria or physical properties, but since MRS medium contains reagents that cannot be used as food materials, cultured lactic acid bacteria can be directly used. It could not be used for food purposes.
これまで、乳酸菌用の培地で、食品素材で構成されたものとしては、大麦焼酎蒸留残液からなる培養基材に糖類、乳化剤を添加した培地が報告されている(特許文献1)が、この培地では乳酸菌の培養による到達菌数が十分ではないという問題があった。 Until now, as a culture medium for lactic acid bacteria made of food materials, a culture medium made of barley shochu distillation residue to which sugars and emulsifiers have been added has been reported (Patent Document 1). There was a problem in that the culture medium did not reach a sufficient number of lactic acid bacteria.
本発明の課題は、食品素材で構成された乳酸菌用の培地であって、乳酸菌の培養による到達菌数が十分となる培地およびそれを利用した技術を提供することである。 An object of the present invention is to provide a medium for lactic acid bacteria that is made of a food material and can reach a sufficient number of bacteria by culturing the lactic acid bacteria, and a technique using the same.
本発明者らは、上記課題を解決するために鋭意研究した結果、大豆ペプチドおよび酵母エキスを含有する培地が、乳酸菌の培養による到達菌数が十分となることを見出し、本発明を完成させた。 As a result of intensive research to solve the above problems, the present inventors found that a medium containing soybean peptide and yeast extract can reach a sufficient number of bacteria by culturing lactic acid bacteria, and completed the present invention. .
すなわち、本発明は、大豆ペプチドおよび酵母エキスを含有する乳酸菌用培地である。 That is, the present invention is a medium for lactic acid bacteria containing soybean peptide and yeast extract.
また、本発明は、乳酸菌を、上記乳酸菌用培地で培養することを特徴とする乳酸菌の培養方法である。 Further, the present invention is a method for culturing lactic acid bacteria, which comprises culturing lactic acid bacteria in the above-mentioned medium for lactic acid bacteria.
更に、本発明は、乳酸菌を、上記乳酸菌用培地で培養して得られることを特徴とする乳酸菌培養物である。 Furthermore, the present invention is a lactic acid bacteria culture obtained by culturing lactic acid bacteria in the above-mentioned medium for lactic acid bacteria.
また更に、本発明は、乳酸菌と、上記の乳酸菌用培地を含有することを特徴とする組成物である。 Furthermore, the present invention is a composition characterized by containing lactic acid bacteria and the above-mentioned medium for lactic acid bacteria.
更にまた、本発明は、上記乳酸菌用培地を用いて培養した乳酸菌菌体を分散媒に分散した後、凍結乾燥を行うことを特徴とする乳酸菌凍結乾燥菌体の製造方法である。 Furthermore, the present invention is a method for producing freeze-dried lactic acid bacteria cells, which comprises dispersing the lactic acid bacteria cells cultured using the above-mentioned medium for lactic acid bacteria in a dispersion medium and then freeze-drying them.
本発明の乳酸菌用培地は、すべての乳酸菌の発育を促し、培養後の到達菌数が高いものとすることができる The medium for lactic acid bacteria of the present invention can promote the growth of all lactic acid bacteria and achieve a high number of bacteria after culture.
しかも、本発明の乳酸菌用培地は、食品素材で構成されているため、培養後、そのまま各種飲食品に利用することができる。 Moreover, since the medium for lactic acid bacteria of the present invention is composed of a food material, it can be used as it is in various foods and drinks after culturing.
本発明の乳酸菌用培地(以下、「本発明培地」という)は、大豆ペプチドおよび酵母エキスを含有するものである。 The medium for lactic acid bacteria of the present invention (hereinafter referred to as "the medium of the present invention") contains soybean peptide and yeast extract.
上記大豆ペプチドは、大豆由来のペプチドであれば特に限定されないが、例えば、大豆を常法に従って酵素、酸等で加水分解してペプチドとしたもの等が挙げられる。この大豆ペプチドの分子量も特に限定されないが、例えば、10000以下程度である。なお、大豆ペプチドの分子量の測定はゲルろ過分析法を用いればよい。また、これら大豆ペプチドは1種または2種以上を用いることができる。これらの大豆ペプチドの中でも、大豆をプロテアーゼ等の酵素で加水分解した大豆ペプトンが好ましい。このような大豆ペプトンとしては、Bacterio-N SS-(B)K(登録商標、マルハニチロ(株))、大豆ペプトン(オルガノフードテック(株))等が市販されている。 The above-mentioned soybean peptide is not particularly limited as long as it is a peptide derived from soybean, but examples include peptides obtained by hydrolyzing soybean with enzymes, acids, etc. according to conventional methods. The molecular weight of this soybean peptide is also not particularly limited, but is, for example, about 10,000 or less. Note that the molecular weight of soybean peptides may be measured using gel filtration analysis. Further, one type or two or more types of these soybean peptides can be used. Among these soybean peptides, soybean peptone obtained by hydrolyzing soybean with an enzyme such as protease is preferred. As such soybean peptone, Bacterio-N SS-(B)K (registered trademark, Maruha Nichiro Co., Ltd.), soybean peptone (Organo Food Tech Co., Ltd.), etc. are commercially available.
本発明培地における大豆ペプチドの含有量は、特に限定されないが、例えば、0.1~20質量%(以下、単に「%」という)、好ましくは0.5~10%、特に好ましくは0.5~5%である。 The content of soybean peptide in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 20% by mass (hereinafter simply referred to as "%"), preferably 0.5 to 10%, particularly preferably 0.5%. ~5%.
上記酵母エキスは、酵母由来のエキスであれば特に限定されないが、例えば、酵母の自己消化液、酵母の抽出液等が挙げられる。酵母の抽出液を得る方法は特に限定されず、例えば、熱水、アルコール等の溶媒で抽出する方法等が挙げられる。酵母エキスは、酵母の自己消化液、酵母の抽出液そのもの、あるいはこれらの濃縮液または粉末等でもよい。酵母の種類は特に限定されず、例えば、ビール酵母、パン酵母、ワイン酵母等が挙げられるが、ビール酵母が好ましい。また、これら酵母エキスは1種または2種以上を用いることができる。これら酵母エキスの中でもビール酵母エキスが好ましい。このようなビール酵母エキスとしては、ミーストP1G(登録商標、アサヒビール食品(株))、ミーストP2G(登録商標、アサヒビール食品(株))、酵母エキス B2(オリエンタル酵母工業(株))等が市販されている。 The above-mentioned yeast extract is not particularly limited as long as it is an extract derived from yeast, and examples thereof include yeast autolytic fluid, yeast extract, and the like. The method for obtaining the yeast extract is not particularly limited, and examples include a method of extraction with a solvent such as hot water or alcohol. The yeast extract may be an autolyzed yeast liquid, a yeast extract itself, or a concentrated liquid or powder thereof. The type of yeast is not particularly limited, and examples thereof include beer yeast, baker's yeast, wine yeast, etc., with beer yeast being preferred. Moreover, these yeast extracts can be used alone or in combination of two or more. Among these yeast extracts, beer yeast extract is preferred. Examples of such brewer's yeast extract include Meest P1G (registered trademark, Asahi Beer Foods Co., Ltd.), Meest P2G (registered trademark, Asahi Beer Foods Co., Ltd.), Yeast Extract B2 (Oriental Yeast Industries Co., Ltd.), etc. It is commercially available.
本発明培地における酵母エキスの含有量は、特に限定されないが、例えば、酵母エキス粉末として0.1~20%、好ましくは0.5~10%である。 The content of yeast extract in the medium of the present invention is not particularly limited, but is, for example, 0.1 to 20% as yeast extract powder, preferably 0.5 to 10%.
本発明培地には、更に発酵大麦エキス、乳化剤および糖類からなる群から選ばれる1種以上、好ましくは全てを含有させてもよい。 The medium of the present invention may further contain one or more selected from the group consisting of fermented barley extract, emulsifiers, and sugars, preferably all of them.
上記発酵大麦エキスは、発酵大麦由来のエキスであれば、特に限定されないが、例えば、大麦を発酵させて焼酎等の酒類を製造する際に得られる、酒類の蒸留後の残液等が挙げられ、具体的には、大麦焼酎蒸留残液等が挙げられる。また、これら発酵大麦エキスは1種または2種以上を用いることができる。これらの発酵大麦エキスの中でも大麦焼酎蒸留残液が好ましい。このような大麦焼酎蒸留残液としては、バーレックス(登録商標、三和酒類(株))、バーレックスS(登録商標、三和酒類(株))等が市販されている。 The above-mentioned fermented barley extract is not particularly limited as long as it is an extract derived from fermented barley, but examples include the residual liquid after distillation of alcoholic beverages obtained when barley is fermented to produce alcoholic beverages such as shochu. , specifically, barley shochu distillation residue and the like. Moreover, these fermented barley extracts can be used alone or in combination of two or more. Among these fermented barley extracts, barley shochu distillation residue is preferred. As such barley shochu distillation residue, Burlex (registered trademark, Sanwa Shurui Co., Ltd.), Burlex S (registered trademark, Sanwa Shurui Co., Ltd.), etc. are commercially available.
本発明培地における発酵大麦エキスの含有量は、特に限定されないが、例えば、エキス(液体)として0.1~50%、好ましくは1.0~25%である。 The content of fermented barley extract in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 50% as extract (liquid), preferably 1.0 to 25%.
上記乳化剤は、特に限定されないが、例えば、モノオレイン酸デカグリセリン、モノステアリン酸デカグリセリン等のポリグリセリン脂肪酸エステルが挙げられる。また、これら乳化剤は1種または2種以上を用いることができる。これら乳化剤の中でもポリグリセリン脂肪酸エステルのうち親水性のポリグリセリン脂肪酸エステルが好ましく、特にモノオレイン酸デカグリセリンが好ましい。このモノオレイン酸デカグリセリンとしては、サンソフトQ-17S(登録商標、太陽化学(株))、NIKKOL DECAGLYN 1-OV(日光ケミカルズ(株))等が市販されている。 The emulsifier is not particularly limited, and examples thereof include polyglycerin fatty acid esters such as decaglycerin monooleate and decaglycerin monostearate. Moreover, these emulsifiers can be used alone or in combination of two or more. Among these emulsifiers, hydrophilic polyglycerol fatty acid esters are preferred among polyglycerol fatty acid esters, and decaglycerin monooleate is particularly preferred. As this decaglycerin monooleate, Sunsoft Q-17S (registered trademark, Taiyo Kagaku Co., Ltd.), NIKKOL DECAGLYN 1-OV (Nikko Chemicals Co., Ltd.), and the like are commercially available.
本発明培地における乳化剤の含有量は、特に限定されないが、例えば、0.001~5%、好ましくは0.01~0.5%である。 The content of the emulsifier in the culture medium of the present invention is not particularly limited, but is, for example, 0.001 to 5%, preferably 0.01 to 0.5%.
上記糖類は、乳酸菌が資化できるものであれば特に限定されないが、例えば、乳糖、グルコース、果糖、ショ糖等が挙げられる。また、これら糖類は1種または2種以上を用いることができる。これら糖類の中でもグルコースおよび/又は乳糖が好ましく、乳糖がより好ましい。乳糖としては乳糖GranuLac200(メグレジャパン(株))、MeggletoseB200(メグレジャパン(株))等が市販されている。グルコースとしては無水結晶ぶどう糖(サンエイ糖化(株))等が市販されている。 The above-mentioned saccharide is not particularly limited as long as it can be assimilated by lactic acid bacteria, and examples thereof include lactose, glucose, fructose, sucrose, and the like. Moreover, these saccharides can be used alone or in combination of two or more. Among these sugars, glucose and/or lactose are preferred, and lactose is more preferred. As lactose, lactose GranuLac200 (Megglet Japan Co., Ltd.), Meggletose B200 (Megglet Japan Co., Ltd.), etc. are commercially available. As glucose, anhydrous crystalline glucose (Sanei Toka Co., Ltd.) and the like are commercially available.
本発明培地における糖類の含有量は、特に限定されないが、例えば、0.1~20%、好ましくは0.5~5%である。 The content of saccharides in the culture medium of the present invention is not particularly limited, but is, for example, 0.1 to 20%, preferably 0.5 to 5%.
更に、本発明培地には、ミネラル分や乳酸菌の増殖を阻害しない成分であって操作性をよくする成分(消泡剤等)等を含有させてもよい。 Furthermore, the culture medium of the present invention may contain minerals, components that do not inhibit the growth of lactic acid bacteria, and which improve operability (antifoaming agents, etc.).
本発明培地の好ましい態様としては、次のものが挙げられる。
大豆ペプチド 0.1~20%、好ましくは0.5~5%
酵母エキス 0.1~20%、好ましくは0.5~10%
発酵大麦エキス 0.1~50%、好ましくは1.0~25%
乳化剤 0.001~5%、好ましくは0.01~0.5%
糖類 0.1~20%、好ましくは0.5~5%
水 残量
Preferred embodiments of the culture medium of the present invention include the following.
Soybean peptide 0.1-20%, preferably 0.5-5%
Yeast extract 0.1-20%, preferably 0.5-10%
Fermented barley extract 0.1-50%, preferably 1.0-25%
Emulsifier 0.001-5%, preferably 0.01-0.5%
Sugars 0.1-20%, preferably 0.5-5%
Water remaining amount
本発明培地の最も好ましい態様としては、以下の組成であり、この組成の培地をSMB培地という。
大豆ペプトン 2%
酵母エキス 2%
発酵大麦エキス 6%
モノオレイン酸デカグリセリン 0.1%
乳糖 2%
水 残量
The most preferred embodiment of the culture medium of the present invention has the following composition, and the culture medium with this composition is referred to as SMB medium.
Soy peptone 2%
Yeast extract 2%
Fermented barley extract 6%
Decaglycerin monooleate 0.1%
Lactose 2%
Water remaining amount
本発明培地は、上記成分を水、乳等の培地基材に、溶解させ、pH調整、殺菌等をすることにより調製することができる。pH調整方法としては水酸化ナトリウム等を添加すればよく、好ましいpHとしては6.0~7.2、より好ましいpHとしては6.2~7.0が挙げられる。なお、本発明に糖類を含有させる場合には、糖類以外を上記基材に溶解させ、殺菌後、糖類を溶解させることが好ましい。 The culture medium of the present invention can be prepared by dissolving the above components in a medium base material such as water or milk, followed by pH adjustment, sterilization, etc. The pH may be adjusted by adding sodium hydroxide or the like, and the preferred pH is 6.0 to 7.2, and the more preferred pH is 6.2 to 7.0. Note that when saccharides are contained in the present invention, it is preferable to dissolve substances other than saccharides in the base material, and then dissolve the saccharides after sterilization.
なお、本発明培地は、培地基材以外の成分を一つに封入し、培地キットとすることもできる。そして、使用時に、この培地キットを適宜基材に溶解させればよい。 Note that the culture medium of the present invention can also be made into a culture medium kit by encapsulating components other than the culture medium base material. Then, at the time of use, this culture medium kit may be appropriately dissolved in a base material.
以上説明した本発明培地は、乳酸菌の培養に用いることができる。本発明培地を用いた乳酸菌の培養条件は特に限定されず、通常の乳酸菌の培養条件を適用することができる。本発明培地で培養できる乳酸菌の種類は特に限定されず、ラクトバチルス属、エンテロコッカス属、ラクトコッカス属、ストレプトコッカス属、ビフィドバクテリウム属の乳酸菌等が挙げられる。これらの乳酸菌の中でもラクトバチルス属の乳酸菌が好ましく、ラクトバチルス属の乳酸菌の中でもラクトバチルス・クリスパータス(Lactobacillus crispatus)、ラクトバチルス・アシドフィルス(Lactobacillus acidophilus)、ラクトバチルス・カゼイ(Lactobacillus casei)、ラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカス(Lactobacillus delbrueckii subsp. bulgaricus)、ラクトバチルス・ヘルベティカス(Lactobacillus helveticus)、ラクトバチルス・デルブルッキー・サブスピーシーズ・ラクティス(Lactobacillus delbrueckii subsp. lactis)、ラクトバチルス・プランタラム(Lactobacillus plantarum)、ラクトバチルス・ガセリ(Lactobacillus gasseri)、ラクトバチルス・アシジピスシス(Lactobacillus acidipiscis)、ラクトバチルス・ブレビス(Lactobacillus brevis)、ラクトバチルス・コリニフォルミス(Lactobacillus coryniformis)、ラクトバチルス・デルブルッキー・サブスピーシーズ・デルブルッキー(Lactobacillus delbrueckii subsp. delbrueckii)、ラクトバチルス・ケフィーリ(Lactobacillus kefiri)、ラクトバチルス・ケフィラノファシエンス・サブスピーシーズ・ケフィラノファシエンス(Lactobacillus kefiranofaciens subsp. kefiranofaciens)、ラクトバチルス・ケフィラノファシエンス・サブスピーシーズ・ケフィリグラナム(Lactobacillus kefranofaciens subsp. kefirgranum)、ラクトバチルス・ノデンシス(Lactobacillus nodensis)、ラクトバチルス・パラブレビス(Lactobacillus parabrevis)、ラクトバチルス・パラカゼイ(Lactobacillus paracasei)、ラクトバチルス・パラケフィーリ(Lactobacillus parakefiri)、ラクトバチルス・ペントーサス(Lactobacillus pentosus)、ラクトバチルス・ぺロレンス(Lactobacillus perolens)、ラクトバチルス・ラムノーサス(Lactobacillus rhamnosus)、ラクトバチルス・サリバリウス(Lactobacillus salivarius)、ラクトバチルス・ツセッティ(Lactobacillus tucceti)、ラクトバチルス・クルバタス(Lactobacillus curvatus)、ラクトバチルス・ジョンソニー(Lactobacillus johnsonii)、ラクトバチルス・ファーメンタム(Lactobacillus fermentum)、ラクトバチルス・マリ(Lactobacillus mali)がより好ましい。これら乳酸菌の1種以上を培養することができる。これらの中でもラクトバチルス・クリスパータス、ラクトバチルス・アシドフィルス、ラクトバチルス・カゼイ、ラクトバチルス・ヘルベティカス、ラクトバチルス・デルブルッキー・サブスピーシーズ・ラクティス、ラクトバチルス・プランタラム、ラクトバチルス・ガセリが好ましく、ラクトバチルス・クリスパータスがより好ましく、ラクトバチルス・クリスパータス YIT 12319(FERM BP-11500、受託日:2011年4月28日、独立行政法人製品評価技術基盤機構特許生物寄託センター(〒292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室))が特に好ましい。 The medium of the present invention described above can be used for culturing lactic acid bacteria. The culture conditions for lactic acid bacteria using the medium of the present invention are not particularly limited, and normal culture conditions for lactic acid bacteria can be applied. The types of lactic acid bacteria that can be cultured in the medium of the present invention are not particularly limited, and include lactic acid bacteria of the genus Lactobacillus, Enterococcus, Lactococcus, Streptococcus, and Bifidobacterium. Among these lactic acid bacteria, lactic acid bacteria of the genus Lactobacillus are preferred, and among the lactic acid bacteria of the genus Lactobacillus, Lactobacillus crispatus , Lactobacillus acidophilus , Lactobacillus casei , and Lactobacillus casei are preferred. Lactobacillus delbrueckii subsp . bulgaricus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp . lactis , Lactobacillus plantarum plantarum ), Lactobacillus gasseri, Lactobacillus acidipiscis, Lactobacillus brevis , Lactobacillus coryniformis , Lactobacillus delbrueckii subsp . Lactobacillus delbrueckii subsp. delbrueckii , Lactobacillus kefiri, Lactobacillus kefiranofaciens subsp. kefiranofaciens , Lactobacillus kefiranofaciens subsp. kefiranofaciens Lactobacillus kefranofaciens subsp. kefirgranum , Lactobacillus nodensis , Lactobacillus parabrevis, Lactobacillus paracasei , Lactobacillus parakefiri , Lactobacillus parakefiri・Lactobacillus pentosus , Lactobacillus perolens, Lactobacillus rhamnosus , Lactobacillus salivarius , Lactobacillus tucceti, Lactobacillus curvatus curvatus ), Lactobacillus johnsonii , Lactobacillus fermentum , and Lactobacillus mali . One or more of these lactic acid bacteria can be cultured. Among these, Lactobacillus crispertus, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus plantarum, and Lactobacillus gasseri are preferred; Lactobacillus crispertus is more preferred, and Lactobacillus crispertus YIT 12319 (FERM BP-11500, date of entrustment: April 28, 2011, Patent Organism Depositary, National Institute of Technology and Evaluation (Kisarazu City, Chiba Prefecture, Japan 292-0818) Kazusa Kamatari 2-5-8 Room 120)) is particularly preferred.
本発明培地で乳酸菌を培養することにより、乳酸菌培養物が得られる。培養温度は乳酸菌によって適宜設定すればよいが、30~37℃が好ましい。本発明培地は、食品素材で構成されているため、この乳酸菌培養物は、そのままあるいは、濃縮、希釈、乾燥等をさせて、従来の飲食品等に配合することができる。また、乳酸菌と、本発明培地を含有する組成物も、同様にして、従来の飲食品等に配合することができる。 By culturing lactic acid bacteria in the medium of the present invention, a lactic acid bacteria culture can be obtained. The culture temperature may be set appropriately depending on the lactic acid bacteria, but 30 to 37°C is preferable. Since the culture medium of the present invention is composed of a food material, the lactic acid bacteria culture can be blended into conventional foods and drinks as is or after being concentrated, diluted, dried, etc. Furthermore, compositions containing lactic acid bacteria and the culture medium of the present invention can be similarly incorporated into conventional foods and drinks.
上記組成物を配合することのできる飲食品としては、特に限定されるものではないが、例えば、ハム、ソーセージ等の食肉加工食品、かまぼこ、ちくわ等の水産加工食品、パン、菓子、バター、ヨーグルトや発酵乳、清涼飲料、乳製品乳酸菌飲料、乳酸菌飲料等が挙げられる。また、飲食品の形態としては、通常用いられる飲食品の形態、例えば、粉末、顆粒等の固体状、ペースト状、液状等が挙げられる。また、微生物乾燥菌体を、錠剤、散剤、チュアブル剤、ハードカプセル剤、ソフトカプセル剤、丸剤、ガム等 に加工してもよい。なお、口腔用組成物として、洗口剤、練歯磨、粉歯磨、水歯磨、口腔用軟膏剤、ゲル剤、錠剤、顆粒剤、細粒剤、グミゼリー、トローチ、タブレット、カプセル、キャンディー、チューインガムなどに、またペットフード等に使用することも可能である。 Foods and drinks to which the above composition can be blended are not particularly limited, but include processed meat foods such as ham and sausages, processed seafood foods such as kamaboko and chikuwa, bread, sweets, butter, and yogurt. Examples include fermented milk, soft drinks, dairy products, lactic acid bacteria drinks, and lactic acid bacteria drinks. In addition, the form of the food/beverage product includes commonly used forms of food/beverage products, such as solid forms such as powder and granules, paste forms, liquid forms, and the like. Furthermore, dried microbial cells may be processed into tablets, powders, chewables, hard capsules, soft capsules, pills, gums, and the like. Oral compositions include mouthwash, toothpaste, powdered toothpaste, water toothpaste, oral ointment, gel, tablet, granule, fine granule, gummy jelly, troche, tablet, capsule, candy, chewing gum, etc. It can also be used in pet food, etc.
本発明培地は、乳酸菌凍結乾燥菌体の製造に利用することが可能である。乳酸菌凍結乾燥菌体を製造する方法は特に限定されず、公知の乳酸菌凍結乾燥菌体を製造する方法を適宜用いることが可能であるが、好ましくは、本発明培地を用いて培養した乳酸菌菌体を分散媒に分散した後、凍結乾燥を行う方法である。 The culture medium of the present invention can be used for producing freeze-dried lactic acid bacteria cells. The method for producing freeze-dried lactic acid bacteria cells is not particularly limited, and any known method for producing freeze-dried lactic acid bacteria cells can be used as appropriate, but preferably lactic acid bacteria cells cultured using the medium of the present invention are used. This is a method in which the product is dispersed in a dispersion medium and then freeze-dried.
上記乳酸菌菌体を分散させる分散媒は乳酸菌の種類に合わせて適宜選択すればよいが、例えば、保護剤および抗酸化剤を含有する水溶液を用いることが好ましい。乳酸菌菌体を分散媒に分散させる方法は特に限定されず、分散媒に乳酸菌菌体を添加し、撹拌等をすればよい。また、分散媒に分散させる乳酸菌菌体の量は特に限定されないが、例えば、0.01~20%程度である。なお、分散させる乳酸菌は基本的に生菌体だが、死菌体が含まれていてもよい。 The dispersion medium in which the lactic acid bacteria cells are dispersed may be appropriately selected depending on the type of lactic acid bacteria, but for example, it is preferable to use an aqueous solution containing a protective agent and an antioxidant. The method of dispersing lactic acid bacteria cells in a dispersion medium is not particularly limited, and may be performed by adding the lactic acid bacteria cells to the dispersion medium and stirring. Further, the amount of lactic acid bacteria cells to be dispersed in the dispersion medium is not particularly limited, but is, for example, about 0.01 to 20%. The lactic acid bacteria to be dispersed are basically live bacteria, but may also contain dead bacteria.
分散媒に用いられる保護剤は、特に限定されず、例えば、グルタミン酸や、グルタミン酸ナトリウム、グルタミン酸カリウム等のグルタミン酸の塩、トレハロース、スクロース、ラクトース、マルトース等の二糖類、グリセロール、マルトデキストリン、サイクロデキストリン、脱脂粉乳、馬鈴薯デンプン等が挙げられる。これら保護剤は1種または2種以上を用いることができるが、トレハロース、脱脂粉乳および馬鈴薯デンプンを用いることが好ましい。分散媒における保護剤の含有量は、特に限定されないが、例えば、1~40%が好ましく、3~40%がより好ましく、3~30%が特に好ましい。 The protective agent used in the dispersion medium is not particularly limited, and includes, for example, glutamic acid, glutamic acid salts such as sodium glutamate and potassium glutamate, disaccharides such as trehalose, sucrose, lactose, and maltose, glycerol, maltodextrin, cyclodextrin, Examples include skimmed milk powder and potato starch. One or more of these protective agents can be used, but it is preferable to use trehalose, skim milk powder, and potato starch. The content of the protective agent in the dispersion medium is not particularly limited, but is preferably, for example, 1 to 40%, more preferably 3 to 40%, particularly preferably 3 to 30%.
分散媒に用いられる抗酸化剤は、特に限定されず、例えば、アスコルビン酸や、アスコルビン酸ナトリウム、アスコルビン酸カルシウム等のアスコルビン酸の塩、ビタミンE、カテキン、グルタチオン、アスタキサンチン等が挙げられる。これら抗酸化剤は1種または2種以上を用いることができるが、アスコルビン酸が好ましい。分散媒における抗酸化剤の含有量は、特に限定されないが、例えば、0.01~10%が好ましく、0.05~5%がより好ましい。 The antioxidant used in the dispersion medium is not particularly limited, and examples include ascorbic acid, salts of ascorbic acid such as sodium ascorbate and calcium ascorbate, vitamin E, catechin, glutathione, and astaxanthin. These antioxidants can be used alone or in combination of two or more, but ascorbic acid is preferred. The content of the antioxidant in the dispersion medium is not particularly limited, but is preferably, for example, 0.01 to 10%, more preferably 0.05 to 5%.
分散媒としては、脱脂粉乳、トレハロース、アスコルビン酸および馬鈴薯デンプンからなる群から選ばれる1種以上を含有する水溶液が好ましく、脱脂粉乳、トレハロース、アスコルビン酸および馬鈴薯デンプンを含有する水溶液がより好ましく、脱脂粉乳、トレハロース、アスコルビン酸および馬鈴薯デンプンをそれぞれ3~40%、3~20%、0.1~5%、3~20%含有する水溶液が特に好ましい。なお、溶解してもpHに影響を与えない分散媒(馬鈴薯デンプン等)については、後述する乳酸菌菌体の分散媒懸濁液のpH調整後に当該分散媒を添加することもできる。 The dispersion medium is preferably an aqueous solution containing one or more selected from the group consisting of skim milk powder, trehalose, ascorbic acid, and potato starch, more preferably an aqueous solution containing skim milk powder, trehalose, ascorbic acid, and potato starch. Particularly preferred are aqueous solutions containing 3-40%, 3-20%, 0.1-5% and 3-20% of milk powder, trehalose, ascorbic acid and potato starch, respectively. In addition, regarding a dispersion medium (potato starch, etc.) that does not affect the pH even when dissolved, the dispersion medium can be added after adjusting the pH of a suspension of lactic acid bacteria in the dispersion medium, which will be described later.
上記のようにして乳酸菌菌体を分散媒に分散した後は、当該分散媒のpHをアルカリ側に調整することが好ましく、pHを7.5以上に調整することがより好ましく、pHを7.5~10.5に調整することがさらに好ましく、pHを8.0~10.0に調整することが最も好ましい。pHの調整は水酸化ナトリウム等のアルカリ物質を用いて行えばよい。 After the lactic acid bacteria cells are dispersed in the dispersion medium as described above, it is preferable to adjust the pH of the dispersion medium to an alkaline side, more preferably to adjust the pH to 7.5 or higher, and adjust the pH to 7.5 or higher. It is more preferable to adjust the pH to 5 to 10.5, and most preferably to adjust the pH to 8.0 to 10.0. The pH may be adjusted using an alkaline substance such as sodium hydroxide.
分散媒のpHを調整した後は、凍結乾燥させる。凍結乾燥の条件は、特に限定されないが、例えば、-35℃~-45℃で6~48時間の凍結処理を行った後、12℃~35℃で40~90時間の乾燥処理を行う条件等を挙げることができる。なお、凍結乾燥機の例としては、TF20-80TANNS((株)宝製作所製)等を挙げることができる。 After adjusting the pH of the dispersion medium, it is freeze-dried. Freeze-drying conditions are not particularly limited, but for example, conditions such as freezing at -35°C to -45°C for 6 to 48 hours followed by drying at 12°C to 35°C for 40 to 90 hours. can be mentioned. An example of the freeze dryer is TF20-80TANNS (manufactured by Takara Seisakusho Co., Ltd.).
斯くして得られる乳酸菌凍結乾燥菌体は、保存後も高い菌数を維持することができる。具体的には、乳酸菌凍結乾燥菌体を37℃で8週間保存後の生菌数が、乳酸菌菌体を分散媒に分散し、当該分散媒のpHを7.0に調整した後、凍結乾燥して得られる乳酸菌凍結乾燥菌体を37℃で8週間保存後の生菌数を100%とした時の120%以上である。なお、当然ながら、上記分散媒のpHを7.0に調整して得られる乳酸菌凍結乾燥菌体の製造には、分散媒のpHを7.5~10.5に調整して得られる乳酸菌凍結乾燥菌体の製造に用いたのと、分散媒のpH以外は同じ条件、菌株を用いる。また、乳酸菌凍結乾燥菌体を37℃で8週間保存後の生残率(%)が、乳酸菌菌体を分散媒に分散し、当該分散媒のpHを7.0に調整した後、凍結乾燥して得られる乳酸菌凍結乾燥菌体を37℃で8週間保存後の生残率を100%とした時の110%以上であることがさらに好ましい。なお、生残率は(保存期間後の生菌数/保存0日の生菌数)×100として計算される。 The freeze-dried lactic acid bacteria cells thus obtained can maintain a high bacterial count even after storage. Specifically, the number of viable bacteria after storing freeze-dried lactic acid bacteria cells at 37°C for 8 weeks is determined by dispersing the lactic acid bacteria cells in a dispersion medium, adjusting the pH of the dispersion medium to 7.0, and then freeze-drying the cells. The number of viable bacteria after storing the freeze-dried lactic acid bacteria cells at 37° C. for 8 weeks is 120% or more. Of course, in order to produce freeze-dried lactic acid bacteria cells obtained by adjusting the pH of the dispersion medium to 7.0, freezing of lactic acid bacteria obtained by adjusting the pH of the dispersion medium to 7.5 to 10.5 is required. The same conditions and strain as used for producing dried bacterial cells are used except for the pH of the dispersion medium. In addition, the survival rate (%) after storing lyophilized lactic acid bacteria cells at 37°C for 8 weeks was determined by dispersing the lactic acid bacteria cells in a dispersion medium, adjusting the pH of the dispersion medium to 7.0, and then lyophilizing the cells after lyophilization. It is more preferable that the survival rate of the freeze-dried lactic acid bacteria cells obtained by the above method is 110% or more when the survival rate after storage for 8 weeks at 37° C. is 100%. The survival rate is calculated as (number of viable bacteria after storage period/number of viable bacteria on day 0 of storage) x 100.
上記の乳酸菌凍結乾燥菌体は、そのまま、あるいは通常食品に添加される他の食品素材と混合することにより、食品や飲料に利用することができる。例えば、食品としては、ハム、ソーセージ等の食肉加工食品、かまぼこ、ちくわ等の水産加工食品、パン、菓子、バター、ヨーグルトや発酵乳等が挙げられる。飲料としては、清涼飲料、乳製品乳酸菌飲料、乳酸菌飲料等が挙げられる。また、飲食品の形態としては、通常用いられる飲食品の形態、例えば、粉末、顆粒等の固体状、ペースト状、液状等が挙げられる。また、微生物乾燥菌体を、錠剤、散剤、チュアブル剤、ハードカプセル剤、ソフトカプセル剤、丸剤、ガム等に加工してもよい。なお、口腔用組成物として、洗口剤、練歯磨、粉歯磨、水歯磨、口腔用軟膏剤、ゲル剤、錠剤、顆粒剤、細粒剤、グミゼリー、トローチ、タブレット、カプセル、キャンディー、チューインガムなどに、またペットフード等に使用することも可能である。 The freeze-dried lactic acid bacteria cells described above can be used in foods and drinks as they are or by mixing them with other food materials that are normally added to foods. For example, foods include processed meat foods such as ham and sausage, processed marine foods such as kamaboko and chikuwa, bread, sweets, butter, yogurt, and fermented milk. Beverages include soft drinks, dairy products lactic acid bacteria drinks, lactic acid bacteria drinks, and the like. In addition, the form of the food/beverage product includes commonly used forms of food/beverage products, such as solid forms such as powder and granules, paste forms, liquid forms, and the like. Furthermore, the dried microbial cells may be processed into tablets, powders, chewable preparations, hard capsules, soft capsules, pills, gums, and the like. Oral compositions include mouthwash, toothpaste, powdered toothpaste, water toothpaste, oral ointment, gel, tablet, granule, fine granule, gummy jelly, troche, tablet, capsule, candy, chewing gum, etc. It can also be used in pet food, etc.
以下、本発明を実施例を挙げて詳細に説明するが、本発明はこれら実施例に何ら限定されるものではない。 EXAMPLES Hereinafter, the present invention will be explained in detail with reference to Examples, but the present invention is not limited to these Examples in any way.
実 施 例 1
培地の調製および乳酸菌の培養:
以下の表1に記載の成分を水に混合、溶解させ、次いで、pHを水酸化ナトリウムで6.2~6.7に調整した後、殺菌して、培地を調製した。これらの培地にラクトバチルス・クリスパータスYIT 12319(FERM BP-11500、受託日:2011年4月28日、独立行政法人製品評価技術基盤機構特許生物寄託センター(〒292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室 )、以下、「LcY」ということもある)を0.25%接種し、37℃で12時間培養した。培養後の生菌数(cfu/ml)およびpHを、それぞれ測定した結果も表1に示した。なお、生菌数の測定は、スパイラルプレーター(EDDY JET、IUL Instruments)で行った。
Implementation example 1
Preparation of medium and culture of lactic acid bacteria:
The components listed in Table 1 below were mixed and dissolved in water, the pH was adjusted to 6.2 to 6.7 with sodium hydroxide, and the mixture was sterilized to prepare a medium. Lactobacillus crispertus YIT 12319 (FERM BP-11500, date of entrustment: April 28, 2011, Japan Patent Organism Depositary, National Institute of Technology and Evaluation (Kazusa, Kisarazu City, Chiba Prefecture, Japan 292-0818) Kamatari 2-5-8 Room 120) (hereinafter also referred to as "LcY") was inoculated at 0.25% and cultured at 37°C for 12 hours. Table 1 also shows the results of measuring the number of viable bacteria (cfu/ml) and pH after culturing. The number of viable bacteria was measured using a spiral plater (EDDY JET, IUL Instruments).
この結果より、培地1~4に比べて、大豆ペプチドと酵母エキスの両方を含む本発明の培地5および6は、培養後の生菌数が高くなることが分かった。 From this result, it was found that media 5 and 6 of the present invention containing both soybean peptide and yeast extract had a higher number of viable bacteria after culture than media 1 to 4.
実 施 例 2
培地の調製および乳酸菌の培養:
以下の表2に記載の成分を用いる以外は、実施例1と同様にして培地を調製した(この培地を「SMB培地」という)。この培地で表3に記載の乳酸菌を0.25%接種し、37℃で12時間または24時間培養した。また、比較としてMRS培地(ベクトンディッキンソン・アンド・カンパニー)を用い、乳酸菌を37℃で12時間または24時間培養した。培養後の生菌数を実施例1と同様にして測定した結果を表3に示した。
Implementation example 2
Preparation of medium and culture of lactic acid bacteria:
A medium was prepared in the same manner as in Example 1 except for using the components listed in Table 2 below (this medium is referred to as "SMB medium"). This medium was inoculated with 0.25% of the lactic acid bacteria listed in Table 3, and cultured at 37°C for 12 or 24 hours. For comparison, lactic acid bacteria were cultured at 37° C. for 12 or 24 hours using MRS medium (Becton Dickinson & Company). The number of viable bacteria after culture was measured in the same manner as in Example 1, and the results are shown in Table 3.
この結果より、SMB培地は、合成培地であるMRS培地と同等あるいは同等以上の生菌数が得られることが分かった。 From this result, it was found that the SMB medium yielded the same or higher number of viable bacteria than the MRS medium, which is a synthetic medium.
実 施 例 3
培地の調製および乳酸菌の培養:
実施例2と同様のSMB培地にB.breve YIT 12272※11またはB.bifidum YIT 10347※12を1%接種し、嫌気条件下、37℃で12時間または24時間培養した。また、比較としてMRS培地(ベクトンディッキンソン・アンド・カンパニー)を用い、乳酸菌を嫌気条件下、37℃で12時間または24時間培養した。培養後の生菌数を実施例1と同様にして測定した結果、B.breve YIT 12272およびB.bifidum YIT 10347もSMB培地に生育することを確認した。
※11:FERM BP-11320、受託日:2010年2月16日、独立行政法人製品評価技術基盤機構特許生物寄託センター(〒292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室 )
※12:FERM BP-10613、受託日:2005年6月23日、独立行政法人製品評価技術基盤機構特許生物寄託センター(〒292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室 )
Implementation example 3
Preparation of medium and culture of lactic acid bacteria:
B. in the same SMB medium as in Example 2. breve YIT 12272 *11 or B. bifidum YIT 10347 *12 was inoculated at 1% and cultured under anaerobic conditions at 37°C for 12 or 24 hours. For comparison, lactic acid bacteria were cultured under anaerobic conditions at 37° C. for 12 or 24 hours using MRS medium (Becton Dickinson & Company). As a result of measuring the number of viable bacteria after culturing in the same manner as in Example 1, B. breve YIT 12272 and B. bifidum YIT 10347 was also confirmed to grow in SMB medium.
*11: FERM BP-11320, Entrusted date: February 16, 2010, Patent Organism Depositary Center, National Institute of Technology and Evaluation (Address: 2-5-8 120 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) Issue room )
*12: FERM BP-10613, date of entrustment: June 23, 2005, National Institute of Technology and Evaluation, Patent Organism Depositary (120 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) Issue room )
実 施 例 4
乳酸菌凍結乾燥菌体の製造:
(1)分散媒懸濁液の調製
SMB培地で前培養したLcYの培養液を、SMB培地2Lに0.1%ずつ接種後、37℃で12時間(定常期に達するまで)静置培養した。次いで、培養液をそれぞれ遠心(4℃、4500×g、7分間)し、菌体を生理食塩水で1回洗浄後、LcYの菌体を分散媒(脱脂粉乳20%、トレハロース10%、アスコルビン酸1%を含有する水溶液)に添加・混合した(pH5.1)。pHを調整しない系では、さらに馬鈴薯デンプンを10%添加・混合し、分散媒懸濁液を得た。一方、分散媒のpHを調整する系では5N水酸化ナトリウムにてpH7.0および9.0にpHを調整した後、馬鈴薯デンプンを10%添加・混合し、分散媒懸濁液を得た。
Implementation example 4
Production of freeze-dried lactic acid bacteria cells:
(1) Preparation of dispersion medium suspension The culture solution of LcY precultured in SMB medium was inoculated into 2 L of SMB medium at 0.1%, and then statically cultured at 37°C for 12 hours (until reaching the stationary phase). . Next, each culture solution was centrifuged (4°C, 4500 x g, 7 minutes), the bacterial cells were washed once with physiological saline, and the LcY bacterial cells were dissolved in a dispersion medium (20% skimmed milk powder, 10% trehalose, ascorbic acid). (pH 5.1). In the system in which the pH was not adjusted, 10% potato starch was further added and mixed to obtain a dispersion medium suspension. On the other hand, in the system for adjusting the pH of the dispersion medium, the pH was adjusted to pH 7.0 and 9.0 with 5N sodium hydroxide, and then 10% potato starch was added and mixed to obtain a dispersion medium suspension.
(2)凍結乾燥および粉砕
(1)で得た分散媒懸濁液を凍結乾燥器(TF20-80TANN:(株)宝製作所製)にてマイナス40℃で2日間凍結後、棚温20℃で2日間凍結乾燥を行った。この凍結乾燥物を粉砕し、LcY凍結乾燥菌体を得た。
(2) Freeze drying and crushing The dispersion medium suspension obtained in (1) was frozen at -40°C for 2 days in a freeze dryer (TF20-80TANN: manufactured by Takara Seisakusho Co., Ltd.), and then at a shelf temperature of 20°C. Freeze-drying was performed for 2 days. This lyophilized product was pulverized to obtain lyophilized LcY cells.
(3)保存試験
(2)で得たLcY凍結乾燥菌体を、チャック付きラミネート袋(ラミジップ:生産日本社製)に分包し、空気をよくぬきチャックを閉めた。その後、37℃にて表3に記載の期間保存し、保存前、保存後の生菌数を測定した。生菌数の測定は、スパイラルプレーター(EDDY JET、IUL Instruments)で行った。また、以下の式で生残率を算出した。さらに、pH無調整の生菌数を100%とした時に、pH7.0または9.0の生菌数との生菌数の比がいくつになるかを以下の式で算出した。それらの結果も表4に示した。
(3) Storage test The LcY freeze-dried cells obtained in (2) were packaged into laminate bags with zippers (LamiZip, manufactured by Seisaku Nippon Sha Co., Ltd.), the air was thoroughly removed, and the zippers were closed. Thereafter, it was stored at 37°C for the period shown in Table 3, and the number of viable bacteria was measured before and after storage. The number of viable bacteria was measured using a spiral plater (EDDY JET, IUL Instruments). In addition, the survival rate was calculated using the following formula. Furthermore, when the number of viable bacteria without pH adjustment was taken as 100%, the ratio of the number of viable bacteria to the number of viable bacteria at pH 7.0 or 9.0 was calculated using the following formula. The results are also shown in Table 4.
この結果より、凍結乾燥前の分散媒のpHを高くすると生菌数や生残率が高くなることが分かった。なお、乳酸菌凍結乾燥菌体の37℃で8週間保存後の生菌数は、pH7.0に調整したものを100%とすると、pH9.0に調整したものは20290%であった。 From this result, it was found that increasing the pH of the dispersion medium before freeze-drying increases the number of viable bacteria and survival rate. In addition, the number of viable bacteria of the freeze-dried lactic acid bacteria cells after storage at 37° C. for 8 weeks was 20290% when the pH was adjusted to 7.0 and 100% when the pH was adjusted to 9.0.
実 施 例 5
乳酸菌凍結乾燥菌体の製造:
実施例4において、静置培養を37℃で12時間行うのに代えて、30℃で22時間行う以外は同様にしてLcY凍結乾燥菌体を得た。これらについて実施例4と同様にして生菌数を測定し、生残率を算出した。また、pH7.0の生菌数を100%とした時に、pH9.0の生菌数との生菌数の比がいくつになるかを以下の式で算出した。その結果を表5に示した。
Implementation example 5
Production of freeze-dried lactic acid bacteria cells:
Lyophilized LcY cells were obtained in the same manner as in Example 4, except that the static culture was performed at 30°C for 22 hours instead of at 37°C for 12 hours. The number of viable bacteria was measured for these in the same manner as in Example 4, and the survival rate was calculated. Furthermore, when the number of viable bacteria at pH 7.0 was taken as 100%, the ratio of the number of viable bacteria to the number of viable bacteria at pH 9.0 was calculated using the following formula. The results are shown in Table 5.
この結果より、培養条件を代えても、凍結乾燥前の分散媒のpHを高くすると生菌数や生残率が高くなることが分かった。 From this result, it was found that even if the culture conditions were changed, increasing the pH of the dispersion medium before freeze-drying increased the number of viable bacteria and survival rate.
実 施 例 6
タンクでの培養:
200Lあるいは1000L容量のステンレスタンクに、SMB培地を160Lあるいは900L入れ、これに前培養したLcYを0.1%接種し、好気条件下、37℃で12時間培養したところ、従来のMRS培地等の培地と比べて同等以上の生菌数が得られた。
Implementation example 6
Culture in tank:
160L or 900L of SMB medium was placed in a stainless steel tank with a capacity of 200L or 1000L, inoculated with 0.1% of precultured LcY, and cultured at 37°C under aerobic conditions for 12 hours. The same or higher number of viable bacteria was obtained compared to the medium.
本発明の乳酸菌用培地は、乳酸菌の培養に用いることができる。 The medium for lactic acid bacteria of the present invention can be used for culturing lactic acid bacteria.
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