JPH11113484A - Improvement of survival of bacillus bifidus - Google Patents
Improvement of survival of bacillus bifidusInfo
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- JPH11113484A JPH11113484A JP28331197A JP28331197A JPH11113484A JP H11113484 A JPH11113484 A JP H11113484A JP 28331197 A JP28331197 A JP 28331197A JP 28331197 A JP28331197 A JP 28331197A JP H11113484 A JPH11113484 A JP H11113484A
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- Prior art keywords
- bifidobacteria
- lactitol
- survival
- culture
- food
- Prior art date
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- Saccharide Compounds (AREA)
- Dairy Products (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、ビフィズス菌の生
残性を改善する方法及びビフィズス菌の含有する飲食物
に関し、特に、発酵乳製品に好適な発明である。TECHNICAL FIELD The present invention relates to a method for improving the survival of bifidobacteria and a food and beverage containing the bifidobacteria, and is particularly suitable for fermented milk products.
【０００２】ここでは、液状の発酵乳製品を主として例
に採り説明するが、発酵乳製品に限られず、飲食物の形
態も、液状、ペースト状、固状のいずれも含む。なお、
ビフィズス菌とは、ビフィドバクテリウム（Bifidobact
erium ）に分類される細菌の一般名である。Here, a liquid fermented dairy product will be mainly described as an example, but the present invention is not limited to a fermented dairy product, and the form of food and drink includes liquid, paste, and solid. In addition,
Bifidobacterium refers to Bifidobacterium
erium) is a generic name for bacteria classified as:
【０００３】[0003]
【従来の技術】ビフィズス菌は、乳幼児にとどまらず、
成人、老人に至るまで、健康に果たす効果は高く評価さ
れている。2. Description of the Related Art Bifidobacteria are not limited to infants,
Its effects on health, from adults to the elderly, are highly valued.
【０００４】例えば、生理的意義に関しては、乳酸、酢
酸、蟻酸等の乳糖起源有機酸の産生にともなう腸管内ｐ
Ｈの低下による病原菌抑制作用、整腸作用等多数の研究
が明らかにされている。このようにビフィズス菌は保健
効果が期待されるため、医薬品や食品に大変多く利用さ
れている。[0004] For example, with regard to the physiological significance, intestinal p
Numerous studies have been clarified, such as an inhibitory action on pathogens due to a decrease in H and an intestinal action. As described above, since bifidobacteria are expected to have health effects, they are widely used in medicines and foods.
【０００５】特に発酵乳（ヨーグルト）等の乳製品に多
く使われ、ビフィズス菌を含んだ各種製品が市場を形成
するに至っている。発酵乳などへの使用菌種としては、
乳幼児にはビフィドバクテリウム・ブレーペ、幼児から
成人用にはビフィドバクテリウム・ロンガムが推奨され
ている（「化学と生物」 vol.24 No.1 ８、９項参
照）。[0005] In particular, various products containing bifidobacteria are widely used in dairy products such as fermented milk (yogurt), and the market has been formed. As bacterial species used for fermented milk, etc.,
Bifidobacterium brepe is recommended for infants and infants, and Bifidobacterium longum is recommended for infants and adults (see “Chemistry and Biology”, vol. 24, No. 18, paragraph 9).
【０００６】[0006]
【発明が解決しようとする課題】しかしながら、ビフィ
ズス菌は従来から発酵乳製造に用いられてきた酪農乳酸
菌と比べ菌学的性質も異なり、下記のような問題点を含
んでいる。However, bifidobacteria differ in mycological properties from dairy lactic acid bacteria conventionally used for the production of fermented milk and have the following problems.
【０００７】生育環境として、酸素が存在する状態で
は生育が遅い偏性嫌気性生菌である。[0007] The growth environment is an obligate anaerobic bacterium that grows slowly in the presence of oxygen.
【０００８】栄養要求性が複雑かつ厳格で酵母エキス
等の生育促進物質を含有しない純粋な牛乳培地では増殖
しない。[0008] The nutritional requirements are complex and strict and do not grow on a pure milk medium containing no growth promoting substances such as yeast extract.
【０００９】耐酸性が低いため、発酵乳のような低ｐ
Ｈ領域で長期間生存させることは困難である。Due to low acid resistance, low p like fermented milk
It is difficult to survive for a long time in the H region.
【００１０】このため、発酵乳製品中でのビフィズス菌
の生菌数に急激な減少が認められ、ビフィズス菌を生き
たまま利用使用という本来の目的を達成することが困難
となる。[0010] For this reason, the viable count of bifidobacteria in fermented milk products is sharply reduced, and it is difficult to achieve the original purpose of utilizing and using bifidobacteria alive.
【００１１】このような観点から発酵乳製品におけるビ
フィズス菌の生残性改善方法について多数研究が行わ
れ、種々提案されている。From such a viewpoint, many studies have been made on methods for improving the survival of bifidobacteria in fermented milk products, and various proposals have been made.
【００１２】例えば、スクロース（ショ糖）またはソル
ビトール（Ｄ−グルシトール）によるビフィズス菌の生
残性改善方法（特公昭５７−４２９１号公報）、エリス
リトールによるビフィズス菌の生残性改善方法（特許第
２５７７６９２号）などがある。For example, a method for improving the survival of bifidobacteria using sucrose (sucrose) or sorbitol (D-glucitol) (Japanese Patent Publication No. 57-4291) and a method for improving the survival of bifidobacteria using erythritol (Japanese Patent No. 2577692) No.).
【００１３】しかし、ショ糖では昨今の低う蝕性（難う
蝕性）や低カロリー化の要請には応え難く、また、ソル
ビトールでは難う蝕性は有するものの低カロリー化の要
請に応え難く、更に、エリスリトールでは上記要請には
応えられるものの、エリスリトールでは、砂糖より甘味
の後味が著しく少なくて、砂糖と同様な甘味感を得難か
った。即ち、砂糖と同様な甘味感が要求される飲食物に
は、砂糖と同様の甘味特性を有する甘味料と併用せざる
を得なかった。However, sucrose is difficult to respond to the recent demand for low cariogenicity (hard caries) and low calories, and sorbitol is hard to meet the demand for low calories although it has hard caries. Although erythritol can meet the above-mentioned requirements, erythritol has a much less sweet aftertaste than sugar, making it difficult to obtain a sweetness similar to that of sugar. In other words, foods and drinks that require a sweetness similar to sugar have to be used in combination with a sweetener having sweetness characteristics similar to sugar.
【００１４】本発明は、上記にかんがみて、従来技術に
優るとも劣らないビフィズス菌の生残性改善ができると
ともに、低カロリー化、低う蝕性の要請に応えられ、し
かも、砂糖と同様な甘味感を容易に得ることができるビ
フィズス菌の生残性改善方法を提供することを目的とす
る。In view of the above, the present invention can improve the survival of bifidobacteria which is not inferior to the prior art, can meet the demand for low calorie and low caries, and has the same effect as sugar. An object of the present invention is to provide a method for improving the survival of bifidobacteria, which can easily obtain a sweet taste.
【００１５】[0015]
【課題を解決するための手段】本発明のビフィズス菌の
生残性改善方法は、上記課題を、飲食物中のビフィズス
菌生残性を改善するために、ラクチトールを、ビフィズ
ス菌の培地または培養物に添加する構成により解決する
ものである。SUMMARY OF THE INVENTION In order to improve the survival of bifidobacteria in foods and drinks, the method for improving the survival of bifidobacteria of the present invention comprises the steps of: The problem is solved by a configuration added to the material.
【００１６】また、本発明のビフィズス菌含有飲食物
は、上記課題を、ビフィズス菌を生残性改善剤と共に含
有する飲食物において、前記生残性改善剤がラクチトー
ルを必須成分とする構成により解決するものである。Further, the food and drink containing bifidobacteria of the present invention solves the above-mentioned problems by providing a food and drink containing bifidobacteria together with a survival improver, wherein the survival improver comprises lactitol as an essential component. Is what you do.
【００１７】更に、本発明のビフィズス菌含有発酵乳製
品の製造方法は、好気性条件下でビフィズス菌を培養し
て得られる培養物そのまま又は適宜加工して、ビフィズ
ス菌を含有する発酵乳製品を製造するに際して、製品１
Ｌ当たり０．０１〜２モル濃度となるように、ラクチト
ールを本培養の培地または培養物に添加混合して製造す
る構成により上記課題を解決するものである。Further, the method for producing a bifidobacterium-containing fermented milk product of the present invention is a method for producing a fermented milk product containing bifidobacteria, which is obtained by culturing the bifidobacterium under aerobic conditions as it is or as appropriate. Product 1 when manufacturing
The object is achieved by a configuration in which lactitol is added to and mixed with a culture medium or a culture for main culture so as to have a concentration of 0.01 to 2 mol per L.
【００１８】[0018]
【発明の構成の説明】以下、本発明の構成について、詳
細に説明をする。なお、以下の説明で「％」、比は、特
に断らない限り、重量単位である。DESCRIPTION OF THE STRUCTURE OF THE INVENTION Hereinafter, the structure of the present invention will be described in detail. In the following description, “%” and the ratio are by weight unless otherwise specified.
【００１９】(1) 本発明の発酵乳製品の培地としては、
通常、発酵乳の製造に用いられている全乳、脱脂乳、ま
たはこれらの粉乳からの還元乳等に適宜生育促進物質な
どを含んだものを使用するが、乳を含まない半合成また
は合成培地を用いることもできる。また、培地の乳固形
分濃度８〜２０％程度では全て使用可能であり、得られ
た培養物は、そのままビフィズス菌を含有する食品とし
て食用に供してもよく、また、甘味料、果汁、水、香料
等を適宜添加し、酪農乳酸菌の発酵乳製品と同様の処理
を行い、飲料としてもよい。(1) The medium of the fermented milk product of the present invention includes:
Usually, whole milk used in the production of fermented milk, skimmed milk, or reduced milk from these milk powders and the like containing a growth promoting substance or the like as appropriate, but a semi-synthetic or synthetic medium containing no milk Can also be used. In addition, all of the medium can be used at a milk solids concentration of about 8 to 20%, and the obtained culture may be used as it is as a food containing bifidobacteria, and may be used as a sweetener, fruit juice, water, or the like. , A fragrance and the like may be added as appropriate, and the same treatment as that for fermented milk products of dairy lactic acid bacteria may be performed to obtain a beverage.
【００２０】なお、上記「合成培地」とは、ＭＲＳ培
地、ブリッグスリバーブロス（Briggsliver broth ）培
地のような、本質的に各種タイプの栄養素・発育因子の
組み合わせにより形成されるものを言う。The above-mentioned "synthetic medium" refers to a medium formed essentially by a combination of various types of nutrients and growth factors, such as an MRS medium and a Briggsliver broth medium.
【００２１】なお、上記培養は、通常好気性条件下で行
う。好気性条件とは、酪農乳酸菌飲料の製造条件での培
養条件をいう。The above culture is usually performed under aerobic conditions. The aerobic condition refers to a culture condition in a dairy lactic acid bacterium beverage production condition.
【００２２】(2) 本発明の発酵乳製品を製造するに際し
て使用するビフィズス菌としては、公知の菌株である、
ビフィドバクテリウム・ブレーベＡＴＣＣ １５７０
０、ビフィドバクテリウム・ロンガムＡＴＣＣ１５７０
７及びビフィドバクテリウム・ブレーベＳＢＲ３２１２
（ＦＡＲＭ Ｐ−１１９１５）等を挙げることができ
る。(2) The bifidobacteria used for producing the fermented milk product of the present invention is a known strain.
Bifidobacterium breve ATCC 1570
0, Bifidobacterium longum ATCC 1570
7 and Bifidobacterium breve SBR3212
(FARM P-11915).
【００２３】(3) 上記ラクチトールは、乳糖のグルコー
ス基が還元されてソルビトール基となった糖アルコール
で、乳糖に水素を添加（還元）して製造される。水素の
添加法は、古くから油脂の硬化に応用され、マーガリン
やショートニング等の製法としてよく知られている慣用
方法で行うことができる。(3) Lactitol is a sugar alcohol in which the glucose group of lactose has been reduced to a sorbitol group, and is produced by adding (reducing) hydrogen to lactose. The method of adding hydrogen has been applied to hardening of fats and oils for a long time, and can be performed by a conventional method well-known as a production method such as margarine or shortening.
【００２４】ラクチトールは、甘味度は低いが（砂糖の
３０〜４０％）、砂糖（ショ糖：スクロース）に似た癖
のない甘味質であり、ヨーロッパにおいては、シュガー
レス菓子関係に多く利用されている。また、水に容易に
溶解し砂糖のように結晶化しやすく、結晶性粉末で吸湿
性がなく、製造上、砂糖と同様の扱いができることが特
徴として挙げられる。また、他の糖アルコールと同じ
く、下記のような特長を有する（社団法人菓子総合技術
センター、「菓子用新素材の適性利用技術シリーズ Ｎ
ｏ．１６」、平成元年３月参照）。Lactitol has a low sweetness (30 to 40% of sugar), but has no habit of sweetness similar to sugar (sucrose: sucrose). In Europe, it is widely used for sugarless confectionery. ing. In addition, it is easily dissolved in water, easily crystallized like sugar, is crystalline powder, has no hygroscopicity, and can be treated in the same manner as sugar in production. Also, like other sugar alcohols, it has the following features (confectionery synthesis technology center, “Appropriate utilization technology series of new materials for confectionery N
o. 16 ", March 1989).
【００２５】消化酵素による分解を受けにくく、小腸
でほとんど吸収されない（エネルギー値は、２kcal/gと
乳糖４kcal/gの半分である。）。It is hardly decomposed by digestive enzymes and is hardly absorbed in the small intestine (the energy value is 2 kcal / g and half that of lactose 4 kcal / g).
【００２６】血糖値がほとんど上がらない。The blood sugar level hardly rises.
【００２７】難う蝕性である。It is hardly carious.
【００２８】また、ラクチトールは、乳酸菌産生菌であ
るBifidobacterium やLactobacill-usには比較的よく利
用され、Bacteriodes やCiostrodium には利用されにく
い。そのため、ラクトースと同等の腸内菌層改善効果が
報告されている（「ビフィズス ９巻１号」 ｐ１９−
２６ １９９５参照）。Lactitol is relatively well used for lactic acid bacteria-producing bacteria, Bifidobacterium and Lactobacill-us, and is hardly used for Bacteriodes and Ciostrodium. Therefore, an intestinal bacterial layer improving effect equivalent to that of lactose has been reported (“Bifidus Vol. 9, No. 1, p. 19-”).
26 1995).
【００２９】しかし、ラクチトールは、全てのビフィズ
ス菌の栄養源となるものではなく、供試菌株であるビフ
ィドバクテリウム・ブレーペに関しては資化されるが、
ビフィドバクテリウム・ロンガムについては資化されな
い。このため、ラクチトールがどのような機構によるも
のかは明らかではないが、ビフィズス菌と共存するだけ
で何らかの保護作用を発揮するのである。However, lactitol is not a nutrient source for all bifidobacteria, and is assimilated with respect to the test strain Bifidobacterium brepe.
Bifidobacterium longum is not assimilated. For this reason, it is not clear what mechanism is used by lactitol, but it exhibits some protective effect only by coexisting with bifidobacteria.
【００３０】従って、ラクチトールはビフィズス菌を含
有する飲食物中に存在させればよい。よって、ラクチト
ールの添加時期は培養前、培養後のいずれの時期でも自
由であるが、培養終了後は、本発明の効果を確実に得る
ためにはなるべく早い段階で加えることが望ましい。Therefore, lactitol may be present in foods and drinks containing bifidobacteria. Therefore, lactitol can be added at any time before or after culturing, but after culturing, it is desirable to add lactitol as early as possible in order to reliably obtain the effects of the present invention.
【００３１】本発明でのラクチトール添加量は、飲食物
の種類、即ち、酸性度及び甘味度により異なる。例え
ば、発酵乳製品の場合、製品１Ｌ当たり０．０１〜２モ
ル、望ましくは０．０１５〜１．７モル、更に望ましく
は、０．０１５〜０．８モルとする。０．０１モル未満
では、生残性改善効果を得難く、２モルを越える使用
は、生残性改善効果が低下する。The amount of lactitol added in the present invention varies depending on the type of food or drink, that is, the acidity and sweetness. For example, in the case of a fermented milk product, the amount is 0.01 to 2 mol, preferably 0.015 to 1.7 mol, more preferably 0.015 to 0.8 mol per liter of the product. If the amount is less than 0.01 mol, it is difficult to obtain a survival improving effect, and if it exceeds 2 mol, the survival improving effect is reduced.
【００３２】[0032]
【発明の作用・効果】本発明のビフィズス菌の生残性改
善方法は、ラクチトールを培地または培養物に添加する
ことにより、下記のような作用・効果を奏する。Action / Effect of the Invention The method for improving the survival of bifidobacteria of the present invention has the following action / effect by adding lactitol to a medium or a culture.
【００３３】これらを用いて得られる飲食物（発酵乳製
品）中の生菌数は、後記実施例に示すように、１０日保
存後でも生菌数低下は少ない。The viable cell count in the food and drink (fermented dairy product) obtained by using them, as shown in the Examples below, shows a small decrease in the viable cell count even after storage for 10 days.
【００３４】即ち、充分商品化可能な程度に生残菌数が
安定し、ビフィズス菌を利用する本来の目的の達成が可
能となり、更に砂糖に似た甘味感を有し、砂糖と同様な
甘味感を容易に得ることができる。That is, the number of surviving bacteria is stable to the extent that it can be sufficiently commercialized, the original purpose of utilizing bifidobacteria can be achieved, and the sweetness similar to sugar is obtained. The feeling can be easily obtained.
【００３５】[0035]
【試験例・応用例】以下、本発明に使用するラクチトー
ルのビフィズス菌の生残性改善効果を確認する為に行っ
た試験例例及び応用例により本発明を説明する。Test Examples and Application Examples The present invention will be described below with reference to test examples and application examples performed to confirm the effect of lactitol used in the present invention to improve the survival of bifidobacteria.
【００３６】なお、各例中の「生菌数」は光岡の嫌気性
用希釈液（光岡：臨床検査、第１８巻、第１１６３頁、
１９７４年）で段階的に希釈した後、血液肝臓寒天（Bl
oodLiver Ager. BL寒天）平板培地の表面に塗布し、３
７℃、７２時間、スチールウール法により嫌気培養を行
い、出現したコロニー数を計測し、試料１ｍＬあたりの
値を示した。また「酸度」は、試料９ｇを中和するのに
要した０．１Ｎ水酸化ナトリウム溶液のｍＬ数により、
試料１ｇあたりの酸度を乳酸％で示した。The "viable cell count" in each case is based on Mitsuoka's anaerobic diluent (Mitsuoka: Clinical Laboratory, Vol. 18, p. 1163,
1974), blood liver agar (Bl
oodLiver Ager. BL agar)
Anaerobic culture was performed by the steel wool method at 7 ° C. for 72 hours, and the number of colonies that appeared was counted, and the value per 1 mL of the sample was shown. The “acidity” is represented by the number of mL of 0.1 N sodium hydroxide solution required to neutralize 9 g of the sample.
The acidity per 1 g of the sample was indicated by% lactic acid.
【００３７】＜試験例１＞０．２％酵母エキス入り１７
％還元脱脂乳培地２００ｍＬを３００ｍＬ三角フラスコ
に分注し、綿栓を施してから、９５℃×３０min の条件
で殺菌した。その後、３７℃まで冷却し、ビフィドバク
テリウム・ブレーベ ATCC15700及び同SBR3212 、ビフィ
ドバクテリウム・ロンガム ATCC15707のスタータを単独
に２％接種し、各３７℃で１８時間静置培養した。得ら
れた培養物を、種々の糖濃度で含有する糖液と培養液を
１：１の割り合いで混合し、５℃で７日間保存した後、
生菌数を測定した。<Test Example 1> 17 containing 0.2% yeast extract
200 mL of% reduced skim milk medium was dispensed into a 300 mL Erlenmeyer flask, covered with a cotton plug, and then sterilized at 95 ° C. for 30 minutes. Thereafter, the mixture was cooled to 37 ° C., and a 2% starter of Bifidobacterium breve ATCC15700, SBR3212, and Bifidobacterium longum ATCC15707 was inoculated alone, and each was allowed to stand still at 37 ° C. for 18 hours. The obtained culture was mixed with a sugar solution containing various sugar concentrations and a culture solution at a ratio of 1: 1 and stored at 5 ° C. for 7 days.
The number of viable bacteria was measured.
【００３８】結果を表１及び図１〜３に示す。各実験に
おいて用いた糖は、糖無添加、ラクチトール、グルコー
ス、スクロースである。なお混合直後の各加糖培養物の
pHは４．４、生菌数は、ビフィドバクテリウム・ブレ
ーベ ATCC15700が４．７×１０9 ／ｍＬ、同SBR3212 が
４．５×１０9 ／ｍＬ、ビフィドバクテリウム・ロンガ
ム ATCC15707が３．７×１０9 ／ｍＬであった。The results are shown in Table 1 and FIGS. The sugars used in each experiment were sugar-free, lactitol, glucose, and sucrose. In addition, immediately after mixing,
The pH was 4.4 and the viable cell count was 4.7 × 10 9 / mL for Bifidobacterium breve ATCC15700, 4.5 × 10 9 / mL for SBR3212, and 3 for Bifidobacterium longum ATCC15707. It was 7 × 10 9 / mL.
【００３９】表１及び図１から、ビフィドバクテリウム
・ブレーベ ATCC15700では、ラクチトール添加により、
０．０２〜１．５モル糖添加濃度のいずれにおいても、
他の糖に比べて生残菌数が高くなる傾向にあった。例え
ば、０．６モル添加での各加糖培養物中の生残菌数は、
ラクチトールが２．１×１０6 ／ｍＬ、グルコースが
１．４Ｅ×１０3 ／ｍＬ、スクロースが６．６×１０4
／ｍＬであった。As shown in Table 1 and FIG. 1, in Bifidobacterium breve ATCC15700, lactitol was added to
At any of the 0.02-1.5 molar sugar addition concentrations,
The number of surviving bacteria tended to be higher than other sugars. For example, the number of surviving bacteria in each sweetened culture at 0.6 mole addition is
Lactitol 2.1 × 10 6 / mL, glucose 1.4E × 10 3 / mL, sucrose 6.6 × 10 4
/ ML.
【００４０】また、同SBR3212 、ビフィドバクテリウム
・ロンガム ATCC15707でも、０．０２〜１．５モル糖添
加濃度のいずれにおいても、他の糖に比べて明らかに生
残菌数が高くなる傾向にあった。例えば、０．６モル添
加での各加糖培養物中の生残菌数は、ビフィドバクテリ
ウム・ブレーベSBR3212 は、ラクチトールが３．８×１
０8 ／ｍＬ、グルコースが１．３３×１０6 ／ｍＬ、ス
クロースが１．６８×１０8 ／ｍＬであった。また、ビ
フィドバクテリウム・ロンガム ATCC15707は、ラクチト
ールが１．５０×１０8 ／ｍＬ、グルコースが９．１×
１０6 ／ｍＬ、スクロースが７．８×１０7 ／ｍＬであ
った。In addition, even in the case of SBR3212 and Bifidobacterium longum ATCC15707, the number of surviving bacteria tended to be clearly higher than that of other sugars at any of the concentrations of 0.02 to 1.5 molar sugar. there were. For example, the number of surviving bacteria in each sweetened culture at the addition of 0.6 mol is as follows: Bifidobacterium breve SBR3212 has 3.8 × 1 lactitol.
0 8 / mL, glucose was 1.33 × 10 6 / mL, and sucrose was 1.68 × 10 8 / mL. In addition, Bifidobacterium longum ATCC15707 contains lactitol at 1.50 × 10 8 / mL and glucose at 9.1 × 10 8 / mL.
It was 10 6 / mL and sucrose was 7.8 × 10 7 / mL.
【００４１】[0041]
【表１】 [Table 1]
【００４２】＜試験例２＞ビフィドバクテリウム・ブレ
ーベ ATCC15700及び同SBR3212 、ビフィドバクテリウム
・ロンガム ATCC15707を増菌用液体培地で、３７℃×２
０時間の条件で培養後、遠心沈殿法により集菌し、数回
洗浄してから菌体懸濁液（ＯＤ660 ＝１．２）を調製し
た。別に種々の０．６モルの糖を含有する各 pH の０．
１モル酢酸緩衝液を調製し、その９ｍＬと上記菌体懸濁
液１ｍＬを混合した後、５℃で保存した。保存３日目、
７日目及び１０日目の生残菌数を測定した。<Test Example 2> Bifidobacterium breve ATCC15700 and SBR3212 and Bifidobacterium longum ATCC15707 were added to a liquid culture medium for enrichment at 37 ° C. × 2.
After culturing for 0 hour, the cells were collected by centrifugation and washed several times to prepare a cell suspension (OD 660 = 1.2). Separately, each of the pH ranges contains 0.6 moles of sugar.
A 1 M acetate buffer was prepared, 9 mL of the buffer was mixed with 1 mL of the cell suspension, and the mixture was stored at 5 ° C. On the third day of storage,
On days 7 and 10, the number of surviving bacteria was measured.
【００４３】結果を示す表２・３から明らかなように、
いずれのビフィズス菌においても各pH での生残菌数の
最も高いのは、ラクチトールであった。特に、 pH ４．
５では顕著であり、その結果を図４〜６に示す。なお、
各図の実験において用いた糖は、糖無添加、スクロー
ス、ラクチトールである。As is clear from Tables 2 and 3 showing the results,
Lactitol had the highest number of surviving bacteria at each pH for all bifidobacteria. In particular, pH4.
5 is remarkable, and the results are shown in FIGS. In addition,
The sugars used in the experiments in each figure are sugar-free, sucrose, and lactitol.
【００４４】図４〜６から明らかなように、保存中のビ
フィドバクテリウム・ブレーベ ATCC15700及び同SBR321
2 、ビフィドバクテリウム・ロンガム ATCC15707の生残
菌数は、糖無添加の場合、保存３日目より生残菌数の減
少が急激に始まり、スクロースを添加した場合、保存３
日目より生残菌数の減少は糖無添加の場合に比して緩や
かである。ラクチトール添加の場合は、スクロース添加
に比べ、更に、生残菌数改善効果があった。As is apparent from FIGS. 4 to 6, Bifidobacterium breve ATCC 15700 and SBR321 during storage.
2. The number of surviving bacteria of Bifidobacterium longum ATCC15707 is rapidly reduced from the third day of storage when no sugar is added.
From the day, the decrease in the number of surviving bacteria is more gradual than in the case without sugar. In the case of adding lactitol, the number of surviving bacteria was further improved as compared with the case of adding sucrose.
【００４５】[0045]
【表２】 [Table 2]
【００４６】[0046]
【表３】 [Table 3]
【００４７】＜応用例１＞ビフィドバクテリウム・ロン
ガム ATCC15707を酵母エキス０．５％を含む１２％還元
脱脂乳培地４００ｍＬに３％接種し、３７℃×１８時間
の条件で培養し、スタータを調製した。このスタータを
直ちに、酵母エキス０．５％を含む１７％還元脱脂乳培
地１０Ｌに３％接種し、３７℃×１８時間の条件で培養
した。ラクチトール、フラクトース、グルコースそれぞ
れについて、表４に示した各糖濃度で含有する糖液と培
養液を１：１の割り合いで混合し、均質機で均質化し
て、発酵乳飲料１０Ｌを調製し、５℃で７日間保存し、
生残菌数を測定した。結果を示す表４から明らかなよう
に、ラクチトールを添加することで、保存製品中の生残
菌の保存が改善された。<Application Example 1> Bifidobacterium longum ATCC15707 was inoculated at a concentration of 3% into 400 mL of a 12% reduced skim milk medium containing 0.5% of yeast extract, and cultured at 37 ° C. for 18 hours. Prepared. This starter was immediately inoculated into 10 L of a 17% reduced skim milk medium containing 0.5% of yeast extract, and cultured at 37 ° C. for 18 hours. For each of lactitol, fructose and glucose, a sugar solution and a culture solution containing at each sugar concentration shown in Table 4 were mixed at a ratio of 1: 1 and homogenized with a homogenizer to prepare 10 L of a fermented milk beverage. Store at 5 ° C for 7 days,
The number of surviving bacteria was measured. As is apparent from Table 4 showing the results, the addition of lactitol improved the preservation of surviving bacteria in the preserved product.
【００４８】[0048]
【表４】 [Table 4]
【００４９】＜応用例２＞０．２％酵母エキス入り１７
％還元脱脂乳１００Ｌを９５℃×３０min の条件で殺菌
した。その後、３７℃まで冷却し、ビフィドバクテリウ
ム・ブレーベSBR3212 のスタータを３％接種し、３７℃
で１８時間静置培養した。ラクチトールが、最終製品で
０．２モルの糖濃度で含有する糖液を培養液を１：１の
割り合いで混合して発酵乳飲料１５０Ｌを調製した。製
品調製直後は、４．５×１０9 ／ｍＬのビフィズス菌を
含有し、 pH は４．７、乳酸酸度０．７％であった。こ
れを５℃で７日間保存した後の生残菌数は、１．２×１
０8 ／ｍＬであった。<Application Example 2> 17 containing 0.2% yeast extract
100 L of 100% reduced skim milk was sterilized at 95 ° C. for 30 minutes. Then, the mixture was cooled to 37 ° C, and 3% of a starter of Bifidobacterium breve SBR3212 was inoculated.
For 18 hours. Lactitol was mixed with a sugar solution containing 0.2 mol of sugar in the final product in a 1: 1 ratio of a culture solution to prepare 150 L of a fermented milk beverage. Immediately after preparation of the product, it contained 4.5 × 10 9 / mL of bifidobacteria, pH was 4.7, and lactic acidity was 0.7%. After storage at 5 ° C. for 7 days, the number of surviving bacteria was 1.2 × 1
Was 0 8 / mL.
【図１】ビフィドバクテリウム・ブレーベ SBR3212の培
養物に各種糖類を添加した場合の糖濃度と７日間保存後
の生残菌数の関係を示すグラフ図である。FIG. 1 is a graph showing the relationship between the sugar concentration and the number of surviving bacteria after storage for 7 days when various sugars are added to a culture of Bifidobacterium breve SBR3212.
【図２】同じくビフィドバクテリウム・ブレーベ ATCC1
5700の培養物における同様な関係を示すグラフ図であ
る。[Figure 2] Bifidobacterium breve ATCC1
FIG. 4 is a graph showing a similar relationship in 5700 cultures.
【図３】同じくビフィドバクテリウム・ロンガム ATCC1
5707の培養物における同様な関係を示すグラフ図であ
る。[Fig. 3] Bifidobacterium longum ATCC1
FIG. 7 is a graph showing a similar relationship in the 5707 culture.
【図４】各糖を含有する pH ４．５の０．１モル酢酸緩
衝液中のビフィドバクテリウム・ブレーベ ATCC15700の
生残菌数と保存日数との関係を示すグラフ図である。FIG. 4 is a graph showing the relationship between the number of surviving bacteria of Bifidobacterium breve ATCC15700 and the number of storage days in a 0.1 M acetate buffer at pH 4.5 containing each sugar.
【図５】同じくビフィドバクテリウム・ブレーベ SBR32
12における同様な関係を示すグラフ図である。Fig. 5 Bifidobacterium breve SBR32
FIG. 13 is a graph showing a similar relationship in FIG.
【図６】同じく同じくビフィドバクテリウム・ロンガム
ATCC15707における同様な関係を示すグラフ図である。FIG. 6: Bifidobacterium longum
FIG. 27 is a graph showing a similar relationship in ATCC15707.
フロントページの続き (72)発明者 山口 勝信 愛知県小牧市小木東三丁目45番地 雪印ロ ーリー株式会社開発研究所内Continuing on the front page (72) Inventor Katsunobu Yamaguchi 3-45 Ogihigashi, Komaki-shi, Aichi Pref.
Claims (4)
るために、ラクチトールを、ビフィズス菌の培地または
培養物に添加することを特徴とするビフィズス菌の生残
性改善方法1. A method for improving the survival of bifidobacteria, which comprises adding lactitol to a culture or culture of bifidobacteria in order to improve the survival of bifidobacteria in foods and drinks.
する飲食物において、前記生残性改善剤がラクチトール
を必須成分とすることを特徴とするビフィズス菌含有飲
食物。2. A food or drink containing bifidobacteria together with a survival improver, wherein the survival improver comprises lactitol as an essential component.
Ｌ当たり０．０１〜２モルのラクチトールを含有するこ
とを特徴とする請求項２記載のビフィズス菌含有飲食
物。3. The food or drink is a fermented milk product, and the product 1
3. The food and drink containing bifidobacteria according to claim 2, which contains 0.01 to 2 mol of lactitol per L.
得られる培養物をそもまま又は適宜加工して、ビフィズ
ス菌を含有する発酵乳製品を製造するに際して、 製品１Ｌ当たり０．０１〜２モル濃度となるように、ラ
クチトールを本培養の培地または培養物に添加混合して
製造することを特徴とするビフィズス菌含有発酵乳製品
の製造方法。4. A method for producing a fermented milk product containing bifidobacteria, wherein a culture obtained by culturing bifidobacteria under aerobic conditions is directly or appropriately processed to produce a fermented dairy product containing bifidobacteria in an amount of 0.01 to 1 L per product. A method for producing a fermented milk product containing bifidobacteria, wherein lactitol is added to and mixed with a medium or a culture for main culture so as to have a 2 molar concentration.
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| JP28331197A JP3261571B2 (en) | 1997-10-16 | 1997-10-16 | Method for improving survival of bifidobacteria |
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|---|---|---|---|---|
| WO2008120570A1 (en) * | 2007-03-19 | 2008-10-09 | National University Corporation Okayama University | Culture medium for production of protein or proliferation of virus |
| US9596875B2 (en) | 2005-06-02 | 2017-03-21 | Kabushiki Kaisha Yakult Honsha | Fermented food containing Bifidobacterium bacteria and method for producing the same |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4881304B2 (en) | 2005-07-21 | 2012-02-22 | 株式会社ヤクルト本社 | Novel Bifidobacterium and its utilization |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9596875B2 (en) | 2005-06-02 | 2017-03-21 | Kabushiki Kaisha Yakult Honsha | Fermented food containing Bifidobacterium bacteria and method for producing the same |
| WO2008120570A1 (en) * | 2007-03-19 | 2008-10-09 | National University Corporation Okayama University | Culture medium for production of protein or proliferation of virus |
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