JPH10327847A - Stabilization of lactobacillus sporogenes spore - Google Patents
Stabilization of lactobacillus sporogenes sporeInfo
- Publication number
- JPH10327847A JPH10327847A JP14224697A JP14224697A JPH10327847A JP H10327847 A JPH10327847 A JP H10327847A JP 14224697 A JP14224697 A JP 14224697A JP 14224697 A JP14224697 A JP 14224697A JP H10327847 A JPH10327847 A JP H10327847A
- Authority
- JP
- Japan
- Prior art keywords
- spores
- spore
- germination
- lactic acid
- lactobacillus sporogenes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【発明が属する技術分野】本発明は、有胞子乳酸菌胞子
の安定化方法に関するものである。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for stabilizing spores of lactic acid bacteria.
【０００２】[0002]
【従来の技術】最近、“体質改善をねらった健康指向の
食品”が世を賑わしており、特に腸の環境改善効果を目
指した商品の要望が高まってきている。そういった中で
ある種の乳酸菌又はビフィズス菌を継続的に飲用するこ
とで腸内菌叢が改善され、それにより健康増進されるこ
とが多くの研究者により研究され知られている。しか
し、乳酸菌は一般に寿命が短く、特に乾燥条件下ではそ
の生存率は極めて低く、また健康増進効果を得るには胃
液及び胆汁酸による影響をのがれて腸管に達する必要性
があり、現在多くのメーカーでは耐酸性乳酸菌の利用、
腸溶性カプセルを用いた導入方法等が考案され用いられ
てきている。一方、有胞子乳酸菌〔バチルス コアグラ
ンス（Bacillus coagulans) 〕は、一般的な乳酸菌と異
なり、一定の条件下で乾燥にも熱や酸にも非常に強く、
腸内で発芽して活発に増殖する特性を有するものであ
り、各種食品用途への応用がなされている。しかしなが
ら、有胞子乳酸菌は、胞子が発芽せずに成熟胞子のまま
でいてこそ、耐熱性等に優れているものであり、一旦発
芽し発芽胞子（発芽胞子は更に栄養細胞へと変化する）
となると、耐熱性、耐薬品性等が失われてしまうことが
知られている。そこで、従来より、有胞子乳酸菌の安定
化（胞子を発芽させずに成熟胞子のまま維持すること）
について各種の検討がなされてきている。有胞子乳酸菌
は粉末の状態では極めて安定であり、１年以上にわたり
一定の菌数を保持することが知られている。しかしなが
ら、水系で安定に保持する方法は未だ提案されてない。
即ち、生理食塩水や糖水溶液に保存しても、経時的に死
滅菌数が増加し、安定に保持しているとは言いがたく、
有胞子乳酸菌の各種飲料等への応用性が損なわれている
のが現状である。2. Description of the Related Art Recently, "health-oriented foods aimed at improving the physical constitution" has been popular in the world, and there has been an increasing demand for products especially aimed at improving the intestinal environment. Under such circumstances, it has been studied and known by many researchers that the continuous ingestion of certain lactic acid bacteria or bifidobacteria improves the intestinal flora and thereby promotes health. However, lactic acid bacteria generally have a short lifespan, and their survival rate is extremely low, especially under dry conditions.In addition, in order to achieve a health-promoting effect, it is necessary to remove the effects of gastric juice and bile acids to reach the intestinal tract. Manufacturers use acid-resistant lactic acid bacteria,
Introduction methods and the like using enteric capsules have been devised and used. On the other hand, spore lactic acid bacteria (Bacillus coagulans), unlike general lactic acid bacteria, are extremely resistant to drying, heat and acid under certain conditions,
It has the property of germinating in the intestine and actively growing, and has been applied to various food applications. However, the spore lactic acid bacterium is excellent in heat resistance and the like only when the spores do not germinate and remain as mature spores, and once germinate and germinate spores (germinative spores are further transformed into vegetative cells)
It is known that heat resistance, chemical resistance and the like are lost. Therefore, conventionally, stabilization of spore-forming lactic acid bacteria (maintaining mature spores without germinating spores)
Various studies have been made on. It is known that spore-forming lactic acid bacteria are extremely stable in a powdered state and maintain a constant number of bacteria for one year or more. However, a method for stably maintaining the compound in an aqueous system has not yet been proposed.
That is, even when stored in saline or aqueous sugar solution, the number of dead sterilization increases with time, and it is hard to say that the bacteria are stably maintained.
At present, the applicability of spore-forming lactic acid bacteria to various beverages and the like has been impaired.
【０００３】[0003]
【課題を解決するための手段】本発明者らは、有胞子乳
酸菌を安定に保持する方法について鋭意検討した結果、
酸性の条件下で発芽抑制剤の添加により、有胞子乳酸菌
の胞子を未発芽の状態で保持することが有効であること
を見出し、本発明を完成するに到った。即ち本発明は、
有胞子乳酸菌に対し、ｐＨが４以下の条件下で発芽抑制
剤を濃度0.5 重量％未満の量添加することにより、有胞
子乳酸菌の胞子を未発芽の状態で保持することを特徴と
する有胞子乳酸菌胞子の安定化方法である。特に好まし
いのは、発芽抑制剤が安息香酸の場合である。本発明で
は、ｐＨと発芽抑制剤の濃度を限定することによって、
食品を腐敗させず、胞子を長期保存することができる。Means for Solving the Problems The present inventors have conducted intensive studies on a method for stably retaining spore-forming lactic acid bacteria.
The present inventors have found that it is effective to keep spores of spore-forming lactic acid bacteria in an ungerminated state by adding a germination inhibitor under acidic conditions, and have completed the present invention. That is, the present invention
Spores of spores of lactic acid bacteria are maintained in an ungerminated state by adding a germination inhibitor to the lactic acid bacteria at a pH of 4 or less at a concentration of less than 0.5% by weight. This is a method for stabilizing lactic acid bacteria spores. Particularly preferred is when the germination inhibitor is benzoic acid. In the present invention, by limiting the pH and the concentration of the germination inhibitor,
The spores can be stored for a long time without spoiling the food.
【０００４】[0004]
【発明の実施の形態】以下、本発明について詳細に説明
する。従来、有胞子細菌の性質として、次のことが知ら
れている。 バチルス コアグランスは、５％の食塩を含有する培
地では生育しない。 バチルス コアグランスは、ｐＨ４以下では生育しな
い。 バチルス メガテリウム（Bacillus megaterium)のリ
ン酸制限培地での生育菌体の胞子は、0.1 ％リン酸添加
培地での胞子よりもかなり耐熱性が高い〔Microbios Le
tt.,20(79)103-107(1982) 〕。 バチルス ステアロサーモフィルス（Bacillus atear
othermophilus)の胞子は、加熱時のリン酸緩衝液の濃度
（０〜67mM）によって胞子の耐熱性が変化し、０mMの時
が最も耐熱性が高い〔Appl.Environ.Microbiol.,36(3)4
57-464(1978)〕。 バチルス コアグランスの胞子は、加熱時のリン酸緩
衝液の濃度によって耐熱性が変化し、10〜25mMの時が最
も耐熱性が高い〔J.Bacteriol.,74:596-604(1957) 〕。 また、胞子の発芽、生育の抑制は、胞子形成の促進と近
似する概念であり、発芽抑制剤としてはヌクレオチド誘
導体（１mM）等が知られている。しかしながら、単にこ
のような発芽抑制剤を添加するのでは、各種飲料等の食
品への利用ができない。また、Mg2+、Ca2+（２mM）は生
育抑制を回復し、胞子形成を阻害すること、並びにアデ
ノシン、無機リン酸（１mM）は胞子の発芽を促進するこ
とが知られている〔Agric.Biol.Chem.,44(12)2773-2778
(1980)、39(9)1893-1895(1975)〕。又、クエン酸ナトリ
ウムなどの有機酸塩が、有胞子細菌の芽胞の発芽を抑制
することも知られている〔食衛誌、25(2)125-131(198
4)〕。更に、〔食品工業、65-71 、２下−1976〕には、
ソルビン酸およびアミノ酸の芽胞発芽阻害性についての
研究が報告されており、バチルス サブチリス（Bacill
us subtilis) PCI-219菌の場合、中性条件下で、食品保
存料（ソルビン酸等）が初期の芽胞発育を阻害すること
が示されている。BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. Conventionally, the following are known as properties of spore-forming bacteria. Bacillus coagulans do not grow on media containing 5% salt. Bacillus coagulans does not grow below pH4. The spores of cells grown on a Bacillus megaterium phosphate-restricted medium are considerably more heat-resistant than spores on a medium supplemented with 0.1% phosphoric acid [Microbios Le.
tt., 20 (79) 103-107 (1982)]. Bacillus stearothermophilus (Bacillus atear)
Othermophilus) spores vary in the heat resistance of spores depending on the concentration of the phosphate buffer during heating (0 to 67 mM), and the spore heat resistance is highest at 0 mM [Appl. Environ. Microbiol., 36 (3) Four
57-464 (1978)]. The heat resistance of spores of Bacillus coagulans changes depending on the concentration of the phosphate buffer at the time of heating, and the heat resistance is highest at 10 to 25 mM [J. Bacteriol., 74: 596-604 (1957)]. In addition, suppression of spore germination and growth is a concept similar to promotion of spore formation, and nucleotide derivatives (1 mM) and the like are known as germination inhibitors. However, simply adding such a germination inhibitor cannot be used for foods such as various beverages. It is known that Mg 2+ and Ca 2+ (2 mM) restore growth inhibition and inhibit spore formation, and adenosine and inorganic phosphate (1 mM) promote spore germination [Agric .Biol.Chem., 44 (12) 2773-2778
(1980), 39 (9) 1893-1895 (1975)]. It is also known that an organic acid salt such as sodium citrate inhibits the germination of spores of spore-forming bacteria (Shokuei, 25 (2) 125-131 (198
Four)〕. In addition, [Food Industry, 65-71, 2-1976]
Studies on the inhibitory effects of sorbic acid and amino acids on spore germination have been reported, and Bacillus subtilis (Bacill
us subtilis) In the case of PCI-219 bacteria, under neutral conditions, food preservatives (such as sorbic acid) have been shown to inhibit early spore development.
【０００５】本発明者らは、上記の公知事実をふまえた
上で鋭意検討した結果、有胞子乳酸菌に対し、ｐＨが４
以下の条件下で発芽抑制剤を濃度0.5 重量％未満の量添
加することにより、有胞子乳酸菌の長期保存条件（加速
試験）での胞子の発芽が顕著に抑制されることを見出し
た。ここで使用する発芽抑制剤とは、安息香酸、ソルビ
ン酸、クエン酸、ヘキサメタリン酸、ピロリン酸、およ
びこれらのカリウム、ナトリウム等の塩である。特に好
ましい発芽抑制剤とは、安息香酸である。The present inventors have conducted intensive studies on the basis of the above-mentioned known facts, and as a result, have found that the pH of the spore-forming lactic acid bacterium is 4
It has been found that spore germination of spore-forming lactic acid bacteria under long-term storage conditions (accelerated test) is significantly suppressed by adding a germination inhibitor in an amount of less than 0.5% by weight under the following conditions. The germination inhibitor used herein is benzoic acid, sorbic acid, citric acid, hexametaphosphoric acid, pyrophosphoric acid, and salts thereof such as potassium and sodium. A particularly preferred germination inhibitor is benzoic acid.
【０００６】本発明において、これらの発芽抑制剤の添
加濃度は、0.5 重量％未満、好ましくは0.2 重量％以
下、特に好ましくは0.01〜0.2 重量％である。また、本
発明においては、ｐＨは４以下、特に４〜３の条件が好
ましい。この範囲外では本発明の効果が得られない。In the present invention, the concentration of these germination inhibitors is less than 0.5% by weight, preferably 0.2% by weight or less, particularly preferably 0.01 to 0.2% by weight. In the present invention, the pH is preferably 4 or less, particularly preferably 4 to 3. Outside this range, the effects of the present invention cannot be obtained.
【０００７】[0007]
【実施例】以下、実施例により本発明を更に具体的に説
明する。 実施例 ラクリス（三共（株）製、登録商標）粉末 0.1ｇを９ml
の無菌水に懸濁し、80℃で30分加熱後、遠心分離するこ
とによって乳糖（賦形剤）などの可溶成分と栄養細胞を
除去すると共に、胞子の活性化処理を行い、再度、無菌
水に懸濁した。予め滅菌した表１に示す各種評価サンプ
ルを添加したＢＣＰ培地（ｐＨ3.8 ）4.9ml に胞子懸濁
液を0.1ml ずつ106 〜107 個／mlとなるように接種後、
50℃に保持した。経時的に0.5ml ずつサンプリングし、
4.5ml の滅菌した50mMリン酸バッファー（ｐＨ７）で希
釈後、80℃で30分加熱後冷却し、ＢＣＰ寒天培地（日水
製薬（株）製）に塗沫し、50℃で２日培養後、生じたコ
ロニー数より胞子の残存率を求めた。結果を表１にあわ
せて示す。EXAMPLES The present invention will be described more specifically with reference to the following examples. Example 9 ml of 0.1 g of Lacris (registered trademark, manufactured by Sankyo Co., Ltd.) powder was used.
Suspended in sterile water, heated at 80 ° C for 30 minutes, centrifuged to remove soluble components such as lactose (excipients) and vegetative cells, and activated spores. Suspended in water. After inoculation previously sterilized spore suspension to various evaluation BCP medium (pH 3.8) with the addition of sample 4.9ml shown in Table 1 so as to have a 0.1ml by 10 6 to 10 7 cells / ml,
It was kept at 50 ° C. Sampling 0.5ml at a time,
After dilution with 4.5 ml of sterile 50 mM phosphate buffer (pH 7), heat at 80 ° C for 30 minutes, cool, spread on BCP agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.), and culture at 50 ° C for 2 days. The remaining rate of spores was determined from the number of colonies formed. The results are shown in Table 1.
【０００８】[0008]
【表１】 [Table 1]
Claims (2)
件下で発芽抑制剤を濃度0.5 重量％未満の量添加するこ
とにより、有胞子乳酸菌の胞子を未発芽の状態で保持す
ることを特徴とする有胞子乳酸菌胞子の安定化方法。1. A method for maintaining spores of a spore-forming lactic acid bacterium in an ungerminated state by adding a germination inhibitor to a spore-forming lactic acid bacterium at a pH of 4 or less at a concentration of less than 0.5% by weight. A method for stabilizing spores of spore-forming lactic acid bacteria.
載の有胞子乳酸菌胞子の安定化方法。2. The method according to claim 1, wherein the germination inhibitor is benzoic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14224697A JPH10327847A (en) | 1997-05-30 | 1997-05-30 | Stabilization of lactobacillus sporogenes spore |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14224697A JPH10327847A (en) | 1997-05-30 | 1997-05-30 | Stabilization of lactobacillus sporogenes spore |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH10327847A true JPH10327847A (en) | 1998-12-15 |
Family
ID=15310852
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14224697A Pending JPH10327847A (en) | 1997-05-30 | 1997-05-30 | Stabilization of lactobacillus sporogenes spore |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH10327847A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018217228A1 (en) * | 2017-05-26 | 2018-11-29 | Muhammed Majeed | Beverage compositions containing bacillus coagulans mtcc 5856 |
-
1997
- 1997-05-30 JP JP14224697A patent/JPH10327847A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018217228A1 (en) * | 2017-05-26 | 2018-11-29 | Muhammed Majeed | Beverage compositions containing bacillus coagulans mtcc 5856 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Desmond et al. | Environmental adaptation of probiotic lactobacilli towards improvement of performance during spray drying | |
| Cotter et al. | Role of the glutamate decarboxylase acid resistance system in the survival of Listeria monocytogenes LO28 in low pH foods | |
| Golden et al. | Inactivation and injury of Listeria monocytogenes as affected by heating and freezing | |
| Mital et al. | Acidophilus milk products: manufacture and therapeutics | |
| Wandling et al. | Nisin in milk sensitizes Bacillus spores to heat and prevents recovery of survivors | |
| Penna et al. | Production of nisin by Lactococcus lactis in media with skimmed milk | |
| JP2000210075A (en) | Lactic acid bacterium having high productivity of gamma- aminobutyric acid, fermented food with high content of gamma-aminobutyric acid using the same lactic acid bacterium and its production | |
| JP3652866B2 (en) | Beverages containing spore lactic acid bacteria | |
| US20020058326A1 (en) | Bifidobacteria and siderophores produced thereby and methods of use | |
| WO2012079973A1 (en) | Food product comprising probiotic bacterial spores | |
| Meera et al. | Partial characterization and optimization of parameters for Bacteriocin production by Probiotic Lactic acid bacteria | |
| Jozala et al. | Increase of nisin production by Lactococcus lactis in different media | |
| Sutherland et al. | Influence of pH and sugars on the growth and production of diarrhoeagenic toxin by Bacillus cereus | |
| JP5133336B2 (en) | Antimicrobial preparation | |
| JPH10327847A (en) | Stabilization of lactobacillus sporogenes spore | |
| Penna et al. | The effect of nisin on growth kinetics from activated Bacillus cereus spores in cooked rice and in milk | |
| US20010019720A1 (en) | Spore germination inhibitor and food using the same | |
| JPH10165173A (en) | Stabilization of spore of bacillus coagulans in aqueous system | |
| JP2001089377A (en) | Agent for suppressing or inhibiting bacterial proliferation containing 1,5-d-anhydrofructose | |
| JP2004337125A (en) | Instant soup containing sporolactobacillus spore | |
| JP2000093162A (en) | Culture of sporogenous lactic acid bacterium | |
| JPH10327846A (en) | Stabilization of lactobacillus sporogenes spore | |
| JP3989626B2 (en) | Method for stabilizing spores of lactic acid bacteria | |
| JPH08205857A (en) | Microorganism protecting agent and production of frozen or frozen and freeze-dried microorganism using the same | |
| EP4379049A1 (en) | Enzyme-containing composition, method for producing milk and method for producing fermented milk |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A131 | Notification of reasons for refusal |
Effective date: 20070109 Free format text: JAPANESE INTERMEDIATE CODE: A131 |
|
| A02 | Decision of refusal |
Effective date: 20070508 Free format text: JAPANESE INTERMEDIATE CODE: A02 |