JP2011177049A - Lactic acid bacterium, beverage using the same and production method thereof - Google Patents
Lactic acid bacterium, beverage using the same and production method thereof Download PDFInfo
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 118
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 59
- 239000004310 lactic acid Substances 0.000 title claims abstract description 59
- 235000013361 beverage Nutrition 0.000 title claims abstract description 56
- 241000894006 Bacteria Species 0.000 title claims abstract description 54
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 23
- 240000002605 Lactobacillus helveticus Species 0.000 claims description 34
- 235000013967 Lactobacillus helveticus Nutrition 0.000 claims description 34
- 229940054346 lactobacillus helveticus Drugs 0.000 claims description 34
- 238000000926 separation method Methods 0.000 claims description 20
- 239000007858 starting material Substances 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 238000012258 culturing Methods 0.000 claims description 17
- 235000013365 dairy product Nutrition 0.000 claims description 15
- 235000020183 skimmed milk Nutrition 0.000 claims description 14
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 8
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 241000186660 Lactobacillus Species 0.000 claims description 5
- 229940039696 lactobacillus Drugs 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 229930024421 Adenine Natural products 0.000 claims description 3
- 229960000643 adenine Drugs 0.000 claims description 3
- GFFGJBXGBJISGV-UHFFFAOYSA-N adenyl group Chemical group N1=CN=C2N=CNC2=C1N GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 3
- 229940104302 cytosine Drugs 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 abstract description 26
- 239000002253 acid Substances 0.000 abstract description 10
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 230000001953 sensory effect Effects 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract 2
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 14
- 238000000855 fermentation Methods 0.000 description 13
- 230000004151 fermentation Effects 0.000 description 13
- 239000000796 flavoring agent Substances 0.000 description 7
- 235000019634 flavors Nutrition 0.000 description 7
- 235000013336 milk Nutrition 0.000 description 6
- 239000008267 milk Substances 0.000 description 6
- 210000004080 milk Anatomy 0.000 description 6
- 239000003381 stabilizer Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 235000013861 fat-free Nutrition 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000002411 adverse Effects 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 235000013618 yogurt Nutrition 0.000 description 3
- 244000199866 Lactobacillus casei Species 0.000 description 2
- 235000013958 Lactobacillus casei Nutrition 0.000 description 2
- 241000186605 Lactobacillus paracasei Species 0.000 description 2
- 244000057717 Streptococcus lactis Species 0.000 description 2
- 235000014897 Streptococcus lactis Nutrition 0.000 description 2
- 229940017800 lactobacillus casei Drugs 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 240000002129 Malva sylvestris Species 0.000 description 1
- 235000006770 Malva sylvestris Nutrition 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 235000014214 soft drink Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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- Dairy Products (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Non-Alcoholic Beverages (AREA)
Abstract
Description
本発明は、中温域で酸生成が速い乳酸菌、その乳酸菌を使用することで、離水や沈殿が抑制された飲料およびそれらの製造方法等に関するものである。 The present invention relates to a lactic acid bacterium that generates acid quickly in a medium temperature range, a beverage in which water separation and precipitation are suppressed by using the lactic acid bacterium, a method for producing the same, and the like.
一般的に、乳酸菌飲料等の飲料は半透明の小さなプラスチック容器で販売されており、製品の沈殿が大きな課題となっている。
しかしながら、乳酸菌飲料等の離水や沈殿は、ベースミックスの殺菌温度と発酵温度（例えば、非特許文献１参照。）、発酵速度（例えば、非特許文献２参照。）、製品ｐＨ等による影響を受け、それらを制御しながらの製造は非常に困難であった。そのため、安定剤の使用により沈殿を抑制している（例えば、特許文献１参照。）。
In general, beverages such as lactic acid bacteria beverages are sold in small translucent plastic containers, and precipitation of products has become a major issue.
However, water separation and precipitation of lactic acid bacteria beverages and the like are affected by the sterilization temperature and fermentation temperature of the base mix (for example, see Non-Patent Document 1), fermentation rate (for example, see Non-Patent Document 2), product pH, and the like. Manufacturing with control of them was very difficult. Therefore, precipitation is suppressed by using a stabilizer (for example, refer to Patent Document 1).
ヨーグルトのスターターとしてはストレプトコッカス・サーモフィルス、ラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリクス、ラクトバチルス・ヘルベティクスといった高温性乳酸菌がポピュラーであり、これらのスターターがさわやかなヨーグルト特有の風味をつくりだしている。しかし、発酵温度は低いほど沈殿、離水を抑制することが知られているため、乳製品乳酸菌飲料、乳酸菌飲料等の製造には高温性乳酸菌がスターターとして利用されることは少ない。 High temperature lactic acid bacteria such as Streptococcus thermophilus, Lactobacillus delbruecki Subspecies bulgaricus, Lactobacillus helveticus are popular as starters of yogurt, and these starters create a refreshing yogurt-specific flavor. . However, since it is known that precipitation and water separation are suppressed as the fermentation temperature is lower, high-temperature lactic acid bacteria are rarely used as starters in the production of dairy lactic acid bacteria beverages, lactic acid bacteria beverages, and the like.
中温域に至適温度をもつ菌としてはラクトコッカス・ラクティス、ラクトバチルス・カゼイ、ラクトバチルス・パラカゼイが知られているが、多くのチーズで利用されているラクトコッカス・ラクティスは慣れ親しんだヨーグルトの味とは異なる風味を呈すことから利用されていなかった。このようなことから、ラクトバチルス・カゼイやラクトバチルス・パラカゼイといった菌種がスターターとして利用されることが多く、風味のバラエティがあまりないのが現状であった。 Lactococcus lactis, Lactobacillus casei, and Lactobacillus paracasei are known as bacteria having the optimal temperature in the middle temperature range, but Lactococcus lactis used in many cheeses is a familiar yogurt taste It was not used because it had a different flavor. For these reasons, bacterial species such as Lactobacillus casei and Lactobacillus paracasei are often used as starters, and there is not much variety in flavor.
沈殿の抑制には製品ｐＨの影響も大きく、長時間発酵させることにより製品ｐＨを乳タンパク質の等電点から離すほど沈殿は抑制される。ｐＨ低下能の高い菌種としてラクトバチルス・ヘルベティクスがあげられるが、本菌は高温性乳酸菌であるため発酵温度の点から沈殿の回避には安定剤を使用する必要があった。 Suppression of precipitation is also greatly affected by the product pH, and precipitation is suppressed as the product pH is separated from the isoelectric point of the milk protein by fermentation for a long time. Lactobacillus helvetics can be mentioned as a species having a high pH lowering ability. However, since this bacterium is a thermophilic lactic acid bacterium, it was necessary to use a stabilizer in order to avoid precipitation from the viewpoint of fermentation temperature.
乳酸菌飲料等の飲料の沈殿は、製品ｐＨ、発酵温度、発酵速度等による影響を受け、それらを制御しながらの製造は非常に困難であった。そのため、安定剤の使用により沈殿を抑制しているが、安定剤の使用は乳酸菌飲料の重要な官能特性であるのどごしやフレーバーリリースに悪影響を及ぼし、さわやかな風味が失われるという問題点もあった。さらに、安定剤を添加するため、コストアップという問題点もあった。
本発明は、これらの問題点を考慮し、飲料等の培養物への離水や沈殿抑制効果を有し、かつ飲料等の培養物の官能特性に悪影響を与えず、製造コストのかからない新規な乳酸菌、これらの乳酸菌を用いた飲料、およびその製造方法等を提供することを課題とする。
Precipitation of beverages such as lactic acid bacteria beverages is affected by product pH, fermentation temperature, fermentation rate, etc., and production while controlling them has been very difficult. For this reason, precipitation is suppressed by the use of stabilizers, but the use of stabilizers has an adverse effect on throat and flavor release, which are important sensory characteristics of lactic acid bacteria beverages, and there is also a problem that a refreshing flavor is lost. . Furthermore, since a stabilizer is added, there is a problem of an increase in cost.
In consideration of these problems, the present invention is a novel lactic acid bacterium that has an effect of inhibiting water separation and precipitation in a culture such as a beverage, does not adversely affect the sensory characteristics of the culture such as a beverage, and does not require production costs. It is an object of the present invention to provide a beverage using these lactic acid bacteria, a production method thereof, and the like.
したがって、本発明は、下記のいずれかの構成からなる発明である。
（１）配列表配列番号１に記載の１６ＳｒＤＮＡ塩基配列と９４％を超える同一性を有し、かつ、６７４番目、１１６５番目の塩基がシトシンであるラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）。
（２）さらに２０２番目の塩基配列がアデニンである請求項１に記載のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）。
（３）還元脱脂乳培地を用いて２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５の範囲である、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）。
（４）ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌ ｖｅｔｉｃｕｓ）ＳＢＴ０６２１（ＦＥＲＭ Ｐ−２１８８３）。
（５）上記（１）乃至（４）のいずれかに記載の乳酸菌を配合した飲料。
（６）前記飲料が、乳製品乳酸菌飲料または乳酸菌飲料である上記（５）に記載の飲料。
（７）上記（１）乃至（４）のいずれかに記載のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）をスターターとして飲料原料に添加して培養する工程を有する飲料の製造方法。
（８）前記培養工程において、２０〜２５℃、６８〜７６時間で培養する上記（７）に記載の飲料の製造方法。
（９）上記（１）乃至（４）のいずれかに記載のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）をスターターとして用いることにより培養物の離水を抑制する方法。
Accordingly, the present invention is an invention having any of the following configurations.
(1) Lactobacillus helveticus which has 94% identity with the 16S rDNA base sequence described in SEQ ID NO: 1 and whose bases at positions 674 and 1165 are cytosine.
(2) The Lactobacillus helveticus according to claim 1, wherein the 202nd base sequence is adenine.
(3) Lactobacillus helveticus having a pH reached in the range of 3.5 to 4.5 when cultured in a reduced skim milk medium at 20 to 25 ° C for 68 to 76 hours.
(4) Lactobacillus helvetics SBT0621 (FERM P-21883).
(5) A beverage containing the lactic acid bacterium according to any one of (1) to (4) above.
(6) The beverage according to (5), wherein the beverage is a dairy lactic acid bacteria beverage or a lactic acid bacteria beverage.
(7) A method for producing a beverage comprising a step of adding a Lactobacillus helveticus (Lactobacillus helveticus) according to any one of (1) to (4) above to a beverage ingredient as a starter and culturing.
(8) The method for producing a beverage according to (7) above, wherein the cultivation is performed at 20 to 25 ° C. for 68 to 76 hours in the culturing step.
(9) A method for suppressing water separation of a culture by using Lactobacillus helveticus according to any one of (1) to (4) as a starter.
中温域で酸生成が速い乳酸菌株を使用することで、沈殿が抑制された飲料等を得ることができる。 By using a lactic acid strain that produces acid fast in the middle temperature range, a beverage or the like in which precipitation is suppressed can be obtained.
本発明者らは、乳酸菌飲料等の沈殿抑制に優れた効果を有し、かつ飲料ののどごしやフレーバーリリースに悪影響を与えない、乳酸菌の製造条件について鋭意検討を進めたところ、中温域で酸生成が速い乳酸菌株の使用により沈殿抑制効果のあることを見出した。
本発明の中温域で酸生成が速い乳酸菌株とは、還元脱脂乳培地を用いて２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５である乳酸菌株のことをいう。さらに、高温域で一晩培養しても乳を凝固させることができない、高温域で酸生成が遅い乳酸菌株であることが好ましい。ここで、高温域で酸生成が遅い乳酸菌株とは、還元脱脂乳培地を用いて３５〜４５℃で１２〜２０時間培養したときの到達ｐＨが４．５〜５．５の範囲である乳酸菌株のことをいう。
The inventors of the present invention have made extensive studies on the production conditions of lactic acid bacteria, which have an excellent effect in suppressing precipitation of lactic acid bacteria beverages and the like, and do not adversely affect the throat and flavor release of beverages. Was found to have a precipitation-inhibiting effect by using a fast lactic acid strain.
The lactic acid strain having a fast acid production in the middle temperature range of the present invention is a lactic acid strain having an ultimate pH of 3.5 to 4.5 when cultured at 20 to 25 ° C. for 68 to 76 hours using a reduced skim milk medium. That means. Furthermore, it is preferable that the strain is a lactic acid strain that cannot coagulate milk even if it is cultured overnight in a high temperature range and has a slow acid production in the high temperature range. Here, a lactic acid bacterium having a slow acid production in a high temperature range is a lactic acid bacterium having a pH reached in the range of 4.5 to 5.5 when cultured at 35 to 45 ° C. for 12 to 20 hours using a reduced skim milk medium. It means a stock.
本発明の乳酸菌の取得方法であるが、例えば自然界に広く分布する乳酸菌の中から、還元脱脂乳培地を用いて２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５である、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を選択することができる。さらに、還元脱脂乳培地を用いて２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５であり、３５〜４５℃で１２〜２０時間培養したときの到達ｐＨが４．５〜５．５の範囲である、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を選択することができる。 In the method for obtaining lactic acid bacteria of the present invention, for example, from among lactic acid bacteria widely distributed in nature, the ultimate pH when culturing at 20 to 25 ° C. for 68 to 76 hours using a reduced skim milk medium is 3.5 to 4 Lactobacillus helveticus, which is .5, can be selected. Furthermore, the ultimate pH when culturing at 20-25 ° C. for 68-76 hours using a reduced skim milk medium is 3.5-4.5, and the ultimate pH when culturing at 35-45 ° C. for 12-20 hours Lactobacillus helveticus can be selected in the range of 4.5 to 5.5.
また、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）の１６ＳｒＤＮＡ塩基配列を調べ、配列表配列番号１に記載の１６ＳｒＤＮＡ塩基配列と９４％を超える同一性を有し、６７４番目および１１６５番目の塩基がシトシンである乳酸菌を選択することにより取得することができる。さらに、２０２番目の塩基配列がアデニンである乳酸菌を選択することにより取得することができる。配列表配列番号１に記載の１６ＳｒＤＮＡ塩基配列との同一性は９４％を超えればよく、特に９７％以上、さらには９９％以上であることが好ましい（微生物の分類・同定実験法、鈴木健一朗編集、シュプリンガー・フェアラーク東京株式会社発行、２００１年９月１６日発行）。 Further, the 16S rDNA base sequence of Lactobacillus helveticus was examined, and it had 94% identity with the 16S rDNA base sequence described in SEQ ID NO: 1 in the sequence listing, and the 674th and 1165th bases were cytosine. It can be obtained by selecting a certain lactic acid bacterium. Furthermore, it can be obtained by selecting a lactic acid bacterium whose 202nd base sequence is adenine. The identity with the 16S rDNA base sequence described in SEQ ID NO: 1 should be more than 94%, preferably 97% or more, more preferably 99% or more (microorganism classification and identification experiment method, edited by Kenichiro Suzuki) , Published by Springer Fairlark Tokyo Co., Ltd., issued September 16, 2001).
このようなラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）として、配列表配列番号２または配列表配列番号３に記載の１６ＳｒＤＮＡ塩基配列を有するラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を例示することができ、特に、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株およびＳＢＴ１００１１株を例示することができる。なお、ＮＣＩＭＢ−７０１２０９株は、Ｎａｔｉｏｎａｌ Ｃｏｌｌｅｃｔｉｏｎｓ ｏｆ Ｉｎｄｕｓｔｒｉａｌ， Ｆｏｏｄ ａｎｄ Ｍａｒｉｎｅ Ｂａｃｔｅｒｉａの公開菌株である。 As such Lactobacillus helvetics, Lactobacillus helveticus having the 16S rDNA base sequence described in SEQ ID NO: 2 or SEQ ID NO: 3 can be exemplified, and particularly Lactobacillus helveticus Examples include Lactobacillus helveticus NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain and SBT10011 strain. The NCIMB-701209 strain is a public strain of National Collections of Industrial, Food and Marine Bacteria.
なお、本発明における選択培地として、還元脱脂乳培地を用いることができるが、例えば、５〜２０％程度の一般的な還元脱脂乳培地を用いることができる。本培地を用いて、２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５であるラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を選択することができる。さらに、本培地を用いて、２０〜２５℃で６８〜７６時間培養したときの到達ｐＨが３．５〜４．５であり、３５〜４５℃で１２〜２０時間培養したときの到達ｐＨが４．５〜５．５の範囲であるラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を選択することができる。 In addition, although a reduced skim milk culture medium can be used as the selective medium in the present invention, for example, a general reduced skim milk medium of about 5 to 20% can be used. Using this medium, Lactobacillus helveticus having an ultimate pH of 3.5 to 4.5 when cultured at 20 to 25 ° C. for 68 to 76 hours can be selected. Furthermore, the ultimate pH when culturing at 20 to 25 ° C. for 68 to 76 hours using this medium is 3.5 to 4.5, and the ultimate pH when culturing at 35 to 45 ° C. for 12 to 20 hours is Lactobacillus helveticus, which is in the range of 4.5 to 5.5, can be selected.
本発明のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を配合して飲料を製造することができる。飲料としては、野菜飲料等の清涼飲料水、茶、機能性飲料、乳製品乳酸菌飲料、乳酸菌飲料等を例示することができるが、特に限定されるものではない。乳製品乳酸菌飲料や乳酸菌飲料等の蛋白質や乳酸菌を多く含む飲料では、特に離水や沈殿抑制の効果が顕著である。 A beverage can be manufactured by blending Lactobacillus helveticus of the present invention. Examples of drinks include soft drinks such as vegetable drinks, tea, functional drinks, dairy lactic acid bacteria drinks, and lactic acid bacteria drinks, but are not particularly limited. In beverages containing a large amount of protein and lactic acid bacteria, such as dairy lactic acid bacteria drinks and lactic acid bacteria drinks, the effects of water separation and precipitation suppression are particularly remarkable.
本発明のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を用いた飲料の製造方法としては、例えば、乳製品乳酸菌飲料や乳酸菌飲料を製造する際に、スターター菌として飲料原料に添加して使用することができる。他のスターター菌と合わせて飲料原料に添加して使用してもよい。本発明のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）をスターターとして飲料原料に添加して培養する工程を有する以外の製造方法は特に変更せずに、離水や沈殿を抑制した飲料を製造することができる。培養温度としては、１５〜２５℃の温度域で培養することにより、さらに、離水や沈殿を抑制することができる。特に２０〜２５℃、６８〜７６時間で培養することが好ましい。原材料や製造装置等は、一般的に飲料製造に用いられるものを用いることができる。 As a method for producing a beverage using Lactobacillus helveticus of the present invention, for example, when producing a dairy lactic acid bacteria beverage or a lactic acid bacteria beverage, it can be used as a starter fungus added to the beverage raw material. it can. You may add and use for a drink raw material in combination with another starter microbe. The production method can be produced without particularly changing the production method other than having a step of adding and culturing the Lactobacillus helveticus of the present invention as a starter to a beverage raw material without any particular change. . As the culture temperature, water separation and precipitation can be further suppressed by culturing in a temperature range of 15 to 25 ° C. In particular, it is preferable to culture at 20 to 25 ° C. for 68 to 76 hours. As the raw material, the manufacturing apparatus, etc., those generally used for beverage production can be used.
本発明のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を用いて培養物を製造することにより、培養物中の離水や沈殿を抑制することができる。後述する発酵試験の結果によると、本発明のラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）を用いて乳製品乳酸菌飲料や乳酸菌飲料を２０〜２５℃で培養して製造した場合、１５℃で２１日間保存後に沈殿や離水はほとんど認められなかった。なお、乳、還元脱脂乳等を原材料として用いた飲料等で特に顕著な効果が認められる。
このように、本発明の乳酸菌スターターを用いることにより、飲料等の培養物の沈殿や離水を抑制することが可能であり、安定剤の添加といったコストアップを回避しつつ、従来の飲料等の培養物の風味を損なわずに飲料等を製造することができる。
By producing a culture using the Lactobacillus helveticus of the present invention, water separation and precipitation in the culture can be suppressed. According to the results of the fermentation test described later, when a dairy lactic acid bacteria beverage or a lactic acid bacteria beverage is cultured at 20 to 25 ° C. using the Lactobacillus helveticus of the present invention and stored at 15 ° C. for 21 days. Later, almost no precipitation or water separation was observed. A particularly remarkable effect is observed in beverages using milk, reduced skim milk or the like as a raw material.
As described above, by using the lactic acid bacteria starter of the present invention, it is possible to suppress precipitation and water separation of a culture such as a beverage, and avoid an increase in cost such as addition of a stabilizer, while culturing a conventional beverage or the like. A drink etc. can be manufactured, without impairing the flavor of a thing.
次に実施例を示し、本発明を詳細に説明する。なお、以下に記載する実施例は本発明を説明するものであり、実施例の記述に限定するものではない。 EXAMPLES Next, an Example is shown and this invention is demonstrated in detail. In addition, the Example described below demonstrates this invention and is not limited to description of an Example.
１０％還元脱脂乳培地に、ラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株および基準株ＪＣＭ１１２０を用いて４０℃で２０時間、２０℃で７６時間培養し、培養後のｐＨを測定した。
それぞれの株の１６ＳｒＤＮＡ塩基配列における２０２番目、６７４番目および１１６５番目の塩基配列と、基準株ＪＣＭ１１２０の１６ＳｒＤＮＡ塩基配列（配列表配列番号１）に対する同一性を表１に示した。また、ＮＣＩＭＢ−７０１２０９株およびＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）の１６ＳｒＤＮＡ塩基配列を配列表配列番号２および３に示した。
Lactobacillus helveticus NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, SBT10011 strain, and reference strain JCM1120 in 10% reduced skim milk medium at 40 ° C. for 20 hours, 20 Culturing was carried out at 76 ° C. for 76 hours, and the pH after the cultivation was measured.
Table 1 shows the identity of the 202S, 674th and 1165th base sequences in the 16S rDNA base sequence of each strain and the 16S rDNA base sequence of the reference strain JCM1120 (SEQ ID NO: 1 in the sequence listing). The 16S rDNA base sequences of NCIMB-701209 and SBT0621 (FERM P-21883) are shown in SEQ ID NOs: 2 and 3, respectively.
結果を表２に示した。基準株ＪＣＭ１１２０の２０℃、７６時間培養後のｐＨは４．７５であり、４０℃、２０時間培養後のｐＨは３．５８であった。一方、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株は２０℃、７６時間培養後のｐＨは３．５５〜３．８７の範囲であり、中温域で酸生成が速く、高温域で酸生成が遅い乳酸菌株であった。また、これらの乳酸菌株はさらに、４０℃、２０時間培養後のｐＨは４．５０〜５．３５の範囲であり、高温域で酸生成が遅い乳酸菌株であった。 The results are shown in Table 2. The pH of the reference strain JCM1120 after culturing at 20 ° C. for 76 hours was 4.75, and the pH after culturing at 40 ° C. for 20 hours was 3.58. On the other hand, NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain have a pH of 3.55 to 3.87 after culturing at 20 ° C. for 76 hours, and acid generation occurs in the middle temperature range. It was a lactic acid strain that was fast and slow in acid production at high temperatures. Further, these lactic acid strains were lactic acid strains having a pH of 4.50 to 5.35 after culturing at 40 ° C. for 20 hours, and slow acid generation in a high temperature range.
１０％還元脱脂乳培地でそれぞれ培養したラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株および基準株ＪＣＭ１１２０をスターターとして用いた。
これらのスターターを９５℃で９０分殺菌した発酵ミックス（１６％脱脂粉乳＋３％グルコース）１０ｇに３％となるように添加し、２５℃で６８時間培養した。この培養物１０ｇを異性化糖４０ｇと混合してＢＲＩＸ １５％の乳製品乳酸菌飲料とした。この乳製品乳酸菌飲料１０ｍｌを目盛り付きのプラスチック容器に入れて１５℃で保存し、７日、１４日、２１日後の離水および沈殿の測定を行った。
Lactobacillus helveticus NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, SBT10011 strain and reference strain JCM1120, each cultured in 10% reduced skim milk medium, were used as starters.
These starters were added to 10 g of a fermented mix (16% nonfat dry milk + 3% glucose) sterilized at 95 ° C. for 90 minutes, and cultured at 25 ° C. for 68 hours. 10 g of this culture was mixed with 40 g of isomerized sugar to give a dairy lactic acid bacteria beverage with 15% BRIX. 10 ml of this dairy lactic acid bacteria beverage was placed in a graduated plastic container and stored at 15 ° C., and water separation and precipitation were measured after 7, 14, and 21 days.
基準株ＪＣＭ１１２０は中温域の２５℃で培養したため、培養後のｐＨは５．１０であり発酵ミックスを凝固せず、乳製品乳酸菌飲料の製造ができなかった。よって、沈殿および離水については経過をみることはできなかった。
一方、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株については、結果を表３に示した。培養後のｐＨは３．５０〜４．５０であり、中温域の２５℃で良好な発酵を示した。さらに、保存中の離水や沈殿がいずれも１ｍｍ以下であり極めて少なく、良質の乳製品乳酸菌飲料が製造できた。
Since the reference strain JCM1120 was cultured at 25 ° C. in the middle temperature range, the pH after the cultivation was 5.10, the fermentation mix was not coagulated, and a dairy lactic acid bacteria beverage could not be produced. Therefore, no progress could be observed for precipitation and water separation.
On the other hand, the results are shown in Table 3 for NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain. The pH after the culture was 3.50 to 4.50, and good fermentation was exhibited at 25 ° C. in the middle temperature range. Furthermore, water separation and precipitation during storage were both 1 mm or less and very little, and a high-quality dairy lactic acid bacteria beverage could be produced.
１０％還元脱脂乳培地でそれぞれ培養したラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）基準株ＪＣＭ１１２０、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株をスターターとした。
このスターターを９５℃で９０分殺菌した発酵ミックス（１６％脱脂粉乳＋３％グルコース）１０ｇに３％となるように添加し、３５℃で２０時間培養した。この培養物１０ｇを異性化糖４０ｇと混合してＢＲＩＸ １５％の乳製品乳酸菌飲料とした。この乳製品乳酸菌飲料１０ｍｌを目盛り付きのプラスチック容器に入れて１５℃で保存し、０日、７日、１４日、２１日後の離水の測定を行った。
Lactobacillus helveticus standard strains JCM1120, NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain each cultured in 10% reduced skim milk medium were used as starters.
This starter was added to 10 g of a fermented mix (16% nonfat dry milk + 3% glucose) sterilized at 95 ° C. for 90 minutes, and cultured at 35 ° C. for 20 hours. 10 g of this culture was mixed with 40 g of isomerized sugar to give a dairy lactic acid bacteria beverage with 15% BRIX. 10 ml of this dairy lactic acid bacteria beverage was placed in a graduated plastic container and stored at 15 ° C., and water separation was measured after 0, 7, 14, and 21 days.
結果を表４に示した。
基準株ＪＣＭ１１２０の培養後のｐＨは３．５０であり、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株の培養後のｐＨは４．５０〜５．２５であった。
保存中の離水の状態であるが、基準株ＪＣＭ１１２０に比べてＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株は離水が顕著に少なかった。なお、実施例２に比べて培養温度が高いため離水が多く認められた。
The results are shown in Table 4.
The pH after culture of the reference strain JCM1120 was 3.50, and the pH after culture of NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain was 4.50 to 5.25. .
Although it was in the state of water separation during storage, NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain had significantly less water separation than the reference strain JCM1120. In addition, since the culture | cultivation temperature was high compared with Example 2, many water separation was recognized.
１０％還元脱脂乳培地でそれぞれ培養したラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）基準株ＪＣＭ１１２０、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株をスターターとした。
このスターターを９５℃で９０分殺菌した発酵ミックス（１６％脱脂粉乳＋３％グルコース）１０ｇに３％となるように添加し、４５℃で１２時間培養した。この培養物１０ｇを異性化糖４０ｇと混合してＢＲＩＸ １５％の乳製品乳酸菌飲料とした。この乳製品乳酸菌飲料１０ｍｌを目盛り付きのプラスチック容器に入れて１５℃で保存し、０日、７日、１４日、２１日後の離水の測定を行った。
Lactobacillus helveticus reference strains JCM1120, NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain, each cultured in 10% reduced skim milk medium, were used as starters.
This starter was added to 10 g of a fermented mix (16% nonfat dry milk + 3% glucose) sterilized at 95 ° C. for 90 minutes, and cultured at 45 ° C. for 12 hours. 10 g of this culture was mixed with 40 g of isomerized sugar to give a dairy lactic acid bacteria beverage with 15% BRIX. 10 ml of this dairy lactic acid bacteria beverage was placed in a graduated plastic container and stored at 15 ° C., and water separation was measured after 0, 7, 14, and 21 days.
結果を表５に示した。基準株ＪＣＭ１１２０の培養後のｐＨは３．５０であった。一方、ＮＣＩＭＢ−７０１２０９株、ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）、ＳＢＴ２１６２株、ＳＢＴ１００１１株の培養後のｐＨは５．２０〜５．５０であり、発酵ミックスを凝固しなかった。 The results are shown in Table 5. The pH of the reference strain JCM1120 after culture was 3.50. On the other hand, pH after cultivation of NCIMB-701209 strain, SBT0621 strain (FERM P-21883), SBT2162 strain, and SBT10011 strain was 5.20-5.50, and the fermentation mix was not coagulated.
１０％還元脱脂乳培地で培養したラクトバチルス・ヘルベティクス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓ）ＳＢＴ０６２１株（ＦＥＲＭ Ｐ−２１８８３）をスターターとした。
このスターターを９５℃で９０分殺菌した発酵ミックス（１６％脱脂粉乳＋３％グルコース）１０ｇに３％となるように添加し、２０℃で７０時間培養した。この培養物５ｇを異性化糖４５ｇと混合してＢＲＩＸ １５％の乳酸菌飲料を製造した。発酵後のｐＨは３．９０であり、良好な発酵を示した。また、保存中の離水や沈殿は抑制されていた。
Lactobacillus helveticus SBT0621 strain (FERM P-21883) cultured in 10% reduced skim milk medium was used as a starter.
This starter was added to 10 g of a fermented mix (16% nonfat dry milk + 3% glucose) sterilized at 95 ° C. for 90 minutes, and cultured at 20 ° C. for 70 hours. 5 g of this culture was mixed with 45 g of isomerized sugar to produce a lactic acid bacteria beverage of 15% BRIX. The pH after fermentation was 3.90, indicating good fermentation. In addition, water separation and precipitation during storage were suppressed.
中温域で酸生成が速い乳酸菌株を使用することで、沈殿が抑制された飲料等を得ることができる。 By using a lactic acid strain that produces acid fast in the middle temperature range, a beverage or the like in which precipitation is suppressed can be obtained.
ＦＥＲＭ Ｐ−２１８８３ FERM P-21883
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