JP5319186B2 - Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition - Google Patents
Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition Download PDFInfo
- Publication number
- JP5319186B2 JP5319186B2 JP2008180659A JP2008180659A JP5319186B2 JP 5319186 B2 JP5319186 B2 JP 5319186B2 JP 2008180659 A JP2008180659 A JP 2008180659A JP 2008180659 A JP2008180659 A JP 2008180659A JP 5319186 B2 JP5319186 B2 JP 5319186B2
- Authority
- JP
- Japan
- Prior art keywords
- lactic acid
- acid bacteria
- food
- ferm
- strain
- 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.)
- Active
Links
Description
本発明は、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５７９菌株、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８０菌株及びＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８１菌株、それを利用して得られた発酵組成物及び漬物食品並びに食品組成物に関する。更に詳しくは、食味を良好にする乳酸菌であるＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５７９菌株、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８０菌株及びＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８１菌株を鮒寿司から単離し、それを利用して得られた発酵組成物及び漬物食品並びに食品組成物に関する。 The present invention relates to a Lactobacillus buchneri FERM P-21579 strain, a Lactobacillus buchneri FERM P-21580 strain and a Lactobacillus buchneri FERM P-21580 strain , a fermented composition and a pickled food obtained using the same, and a food composition. More specifically, Lactobacillus buchneri FERM P-21579 strain, Lactobacillus buchneri FERM P-21580 strain and Lactobacillus buchneri FERM P-21580 strain, which are lactic acid bacteria that improve the taste, were obtained from sushi and obtained by using it. The present invention relates to fermented compositions, pickled foods, and food compositions.
乳酸菌は、自然界に広く分布しており、古くから意図的あるいは非意図的に、食品の風味及び保存性を高めるために用いられてきた。また、乳酸菌は日本の発酵食品とも馴染み深い細菌であり、漬物や味噌にも存在し、食品保存や風味の工夫を重ねる過程を通じて、乳酸菌が意図的あるいは非意図的に選択等されてきた。また、世界各地で植物、乳及び肉などの原料に基づいて、様々な発酵食品の製造に用いられ、その土地及び風土に根付いた多様な発酵食品が作られている。乳酸菌は、現在では、ヨーグルトや乳酸飲料などの発酵乳製品、キムチ、ピクルスなどの発酵植物製品、発酵肉、醸造及び発酵豆乳など多くの飲食品に利用されている（例えば、下記特許文献１参照）。 Lactic acid bacteria are widely distributed in nature and have been used for a long time intentionally or unintentionally to enhance the flavor and preservation of food. In addition, lactic acid bacteria are familiar bacteria with Japanese fermented foods and are also present in pickles and miso. Lactic acid bacteria have been selected intentionally or unintentionally through the process of preserving food and flavoring. In addition, various fermented foods that are used in the manufacture of various fermented foods based on raw materials such as plants, milk, and meat have been made in various parts of the world. Lactic acid bacteria are currently used in many foods and beverages such as fermented dairy products such as yogurt and lactic acid beverages, fermented plant products such as kimchi and pickles, fermented meat, brewing and fermented soy milk (for example, see Patent Document 1 below). ).
近年、乳酸菌及びその発酵食品による健康維持に対する効果が注目されており、更に、食味や機能性を向上させる乳酸菌及びその食品等の開発が求められている。
本発明は、上記の課題を解決するものであり、乳酸菌の分離源として日本の伝統食品の１つである食味良好な鮒寿司に着目した。鮒寿司から単離した乳酸菌が食味を向上させることを見出し、単離した乳酸菌を同定すると共にそれを発酵して得られる発酵組成物、及び漬物食品並びに食品組成物を提供することを目的とする。
In recent years, attention has been focused on the health maintenance effect of lactic acid bacteria and their fermented foods, and further development of lactic acid bacteria and their foods that improve the taste and functionality is required.
The present invention solves the above-mentioned problems, and has focused attention on sushi with good taste, which is one of the traditional Japanese foods as a source for separating lactic acid bacteria. An object of the present invention is to provide a fermented composition obtained by fermenting a lactic acid bacterium isolated from sushi, identifying the isolated lactic acid bacterium and fermenting it, and a pickled food and a food composition .
本発明は以下のとおりである。
１．鮒寿司から単離され、独立行政法人産業技術総合研究所特許生物寄託センター受託番号ＦＥＲＭ Ｐ−２１５７９であることを特徴とするＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５７９菌株。
２．鮒寿司から単離され、独立行政法人産業技術総合研究所特許生物寄託センター受託番号ＦＥＲＭ Ｐ−２１５８０であることを特徴とするＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８０菌株。
３．鮒寿司から単離され、独立行政法人産業技術総合研究所特許生物寄託センター受託番号ＦＥＲＭ Ｐ−２１５８１であることを特徴とするＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８１菌株。
４．上記１．乃至３．の少なくともいずれか一項に記載の菌株を利用して得られたことを特徴とする発酵組成物。
５．上記１．乃至３．の少なくともいずれか一項に記載の菌株を添加して発酵させることにより得られることを特徴とする漬物食品。
６．上記１．乃至３．の少なくともいずれか一項に記載の菌株を培養して得られる菌体を含むことを特徴とする食品組成物。
The present invention is as follows.
1. Lactobacillus buchneri FERM P-21579 strain isolated from sushi and having the accession number FERM P-21579 of the National Institute of Advanced Industrial Science and Technology.
2. Lactobacillus buchneri FERM P-21580 strain isolated from sushi and having the accession number FERM P-21580, National Institute of Advanced Industrial Science and Technology.
3. A Lactobacillus buchneri FERM P-21581 strain isolated from sushi and having the accession number FERM P- 21581 of the National Institute of Advanced Industrial Science and Technology.
4). Above 1. To 3. A fermentation composition obtained by using the strain described in any one of the above.
5. Above 1. To 3. A pickled food obtained by adding and fermenting the strain according to any one of the above.
6). Above 1. To 3. A food composition comprising cells obtained by culturing the strain according to any one of the above.
本発明のＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５７９菌株、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８０菌株及びＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８１菌株は鮒寿司から単離された新規の乳酸菌である。
また、本発明の菌株は食品の風味と保存性を高めることができる。
また、本発明の菌株を利用して得られた発酵組成物は優れた食味を奏することができる。
また、本発明の漬物食品は、本発明の菌株を漬物用スターターとして利用することで、食味が安定して良好であり、且つ保存性に優れる漬物食品とすることができる。
更に、本発明食品組成物は、本発明の菌株を培養して得られる菌体を含有させることにより食味が安定し、且つ食品の風味に優れる食品組成物とすることができる。
The Lactobacillus buchneri FERM P-21579 strain, Lactobacillus buchneri FERM P-21580 strain and Lactobacillus buchneri FERM P-21580 strain of the present invention are novel lactic acid bacteria isolated from sushi.
Moreover, the strain of the present invention can enhance the flavor and storage stability of food.
Moreover, the fermented composition obtained using the strain of the present invention can exhibit an excellent taste.
Moreover, the pickled food of the present invention can be made into a pickled food having a stable and good taste and excellent storage stability by using the strain of the present invention as a starter for pickled.
Furthermore, the food composition of the present invention can be made into a food composition having a stable taste and excellent food flavor by containing the cells obtained by culturing the strain of the present invention.
以下、本発明を詳しく説明する。
本発明における新規乳酸菌（以下、単に「乳酸菌」ともいう。）である、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５７９菌株、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８０菌株及びＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉ ＦＥＲＭ Ｐ−２１５８１菌株は、鮒寿司から単離された３種類の乳酸菌である。この乳酸菌の分離源として、滋賀県高島市勝野１２８７にある「喜多品老舗」より鮒寿司を入手し、ストマッカーで均質にした一部を乳酸菌分離用のＭＲＳ白亜寒天培地で培養した。
上記鮒寿司より単離した３種類の新規乳酸菌は、平成２０年５月２２日に本願出願人によって、独立行政法人産業技術総合研究所へ寄託し、受託番号：「ＦＥＲＭ Ｐ−２１５７９」、「ＦＥＲＭ Ｐ−２１５８０」及び「ＦＥＲＭ Ｐ−２１５８１」を得た。
尚、この３種類の新規乳酸菌を以下、受託番号：ＦＥＲＭ Ｐ−２１５７９は「ＳＵ−２」及び受託番号：ＦＥＲＭ Ｐ−２１５８０は「ＳＵ−４」並びに受託番号：ＦＥＲＭ Ｐ−２１５８１は「ＳＵ−６」と略記する。
The present invention will be described in detail below.
Lactobacillus buchneri FERM P-21579 strain, Lactobacillus buchneri FERM P-21580 strain and Lactobacillus buchneri FERM P-21580 strain, which are novel lactic acid bacteria in the present invention (hereinafter also simply referred to as “lactic acid bacteria”), There are three types of lactic acid bacteria. As a source for the separation of lactic acid bacteria, sushi was obtained from “Kita product long-established store” in 1287 Katsuno, Takashima City, Shiga Prefecture, and a portion homogenized with a stomacher was cultured on an MRS chalk agar medium for separating lactic acid bacteria.
Three types of new lactic acid bacteria isolated from the sushi sushi were deposited on May 22, 2008 by the applicant of the present application with the National Institute of Advanced Industrial Science and Technology, with accession numbers: “FERM P-21579”, “ FERM P-21580 "and" FERM P-21580 "were obtained.
In addition, these three kinds of new lactic acid bacteria are hereinafter referred to as “SU-2” for the accession number: FERM P-21579, “SU-4” for the accession number: FERM P-21580, and “SU-” for the accession number: FERM P-21580. 6 ”.
本発明の新規乳酸菌を培養する培地としては、表１に示す組成物からなるＭＲＳ培地（以下、「ＭＲＳ培地」と略記する。）が好ましい。 As a medium for culturing the novel lactic acid bacteria of the present invention, an MRS medium comprising the composition shown in Table 1 (hereinafter abbreviated as “MRS medium”) is preferable.
本発明における上記３種の新規乳酸菌は、以下の性質を有する。
（１）形態
上記乳酸菌「ＳＵ−２」及び「ＳＵ−４」並びに「ＳＵ−６」は、全て桿状である。菌の大きさは、巾：０．４〜１．０μｍ、長さ：０．８〜３μｍであり、グラム染色は全て陽性である。
（２）生理学的性質
上記乳酸菌「ＳＵ−２」及び「ＳＵ−４」並びに「ＳＵ−６」の生理学的性質について、「コロニー性状」、「好気培養」、「カタラーゼ」、「１５℃の生育」、「グルコースからのガス産生」及び「糖の発酵性の有無」を表２に示す。尚、表中において陽性は「＋」、及び陰性は「−」で表示した（以下、同様）。
尚、糖の発酵性の有無は以下の方法で行った。
（ａ）糖の発酵性の試験培地は、上記のＭＲＳ培地の糖濃度を０．５質量％に変更し、更にＬ−ｃｙｓｔｅｉｎ−ＨＣｌ−Ｈ２Ｏを０．０２質量％、ｐＨ指示薬としてブロモクレゾールフェノール（Ｂ．Ｃ．Ｐ）を０．０４質量％となるように加えた。また、ｐＨを７．２に調製し、１１５℃で２０分間、高圧滅菌をおこなった。
（ｂ）白金耳でＭＲＳ寒天培地に画線して３３℃、４８時間嫌気培養する。
（ｃ）コロニーを白金耳でそれぞれ釣菌してＭＲＳ培地６ｍｌに接種し、３３℃、２４時間嫌気培養する。
（ｄ）培養液２００μｌをＭＲＳ培地６ｍｌに接種し、１９時間嫌気培養する。
（ｅ）菌体を３０００ｒｐｍ、１５分間遠心分離した後、０．１％Ｌ−ｃｙｓｔｅｉｎ・ＨＣＬ・Ｈ２Ｏと０．１％チオグリコール酸ナトリウムとを加えた滅菌生食水２．５ｍｌで原菌液の２．４倍に懸濁したものを接種菌液とする。
（ｆ）上記菌液をチップで吸い取り、各試験培地（５ｍｌ）の底部に１２５μｌづつ接種する。
（ｇ）上記菌液を接種した試験培地を３３℃で、７日間培養し、糖の発酵性の結果を調べた。
The three kinds of novel lactic acid bacteria in the present invention have the following properties.
(1) Forms The lactic acid bacteria “SU-2”, “SU-4” and “SU-6” are all in the form of a bowl. The size of the bacteria is width: 0.4 to 1.0 μm, length: 0.8 to 3 μm, and Gram staining is all positive.
(2) Physiological properties Regarding the physiological properties of the lactic acid bacteria “SU-2” and “SU-4” and “SU-6”, “colony properties”, “aerobic culture”, “catalase”, “15 ° C. Table 2 shows the “growth”, “gas production from glucose”, and “presence / absence of fermentability of sugar”. In the table, positive is indicated by “+” and negative is indicated by “−” (hereinafter the same).
In addition, the presence or absence of fermentability of sugar was performed by the following method.
(A) The sugar fermentability test medium is prepared by changing the sugar concentration of the MRS medium to 0.5% by mass, further adding 0.02% by mass of L-cysteine-HCl-H 2 O and bromo as a pH indicator. Cresol phenol (BCP) was added so that it might become 0.04 mass%. Moreover, pH was adjusted to 7.2, and high-pressure sterilization was performed at 115 ° C. for 20 minutes.
(B) Streaking on MRS agar medium with platinum ears and anaerobic culture at 33 ° C. for 48 hours.
(C) Each colony is fished with a platinum loop and inoculated into 6 ml of MRS medium, followed by anaerobic culture at 33 ° C. for 24 hours.
(D) 200 μl of the culture solution is inoculated into 6 ml of MRS medium and cultured anaerobically for 19 hours.
(E) After centrifuging the cells at 3000 rpm for 15 minutes, the stock bacterial solution was diluted with 2.5 ml of sterile saline containing 0.1% L-cysteine / HCL / H 2 O and 0.1% sodium thioglycolate. 2.4 Inoculate the suspension with an inoculum.
(F) The above bacterial solution is sucked up with a chip, and 125 μl is inoculated at the bottom of each test medium (5 ml).
(G) The test medium inoculated with the above bacterial solution was cultured at 33 ° C. for 7 days, and the results of sugar fermentability were examined.
本発明の乳酸菌における１６ＳリボソームＲＮＡ塩基配列の解析を以下の６本のプライマーを使用し、シーケンスを行い、ＢＬＡＳＴ検索した。
また、理化学研究所微生物系統保存事業（ＪＣＭ：Ｊａｐａｎ Ｃｏｌｌｅｃｔｉｏｎ ｏｆ Ｍｉｃｒｏｏｒｇａｎｉｓｍｓ）からＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉの標準株ＪＣＭ１１１５を取り寄せて１６ＳｒＲＮＡ遺伝子配列を決定し、分離株（ＳＵ−２、ＳＵ−４、ＳＵ−６）のそれぞれの１６ＳｒＲＮＡ遺伝子配列と比較した。その結果、これらの分離株の配列は標準株とほぼ１００％一致し、これらの分離株はＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉと同定された。なお、上記標準株の１６ＳｒＲＮＡの遺伝子配列はＮＣＢＩ Ｇｅｎｅ Ｂａｎｋに登録した（ＡＣＣＥＳＳＩＯＮ 番号；ＡＢ２０５０５５）。
１６ＳｒＲＮＡ塩基解析に使用したプライマーは以下の通り。尚、下記プライマーの内（１）と（６）はＴｈｅ Ｅｕｒｏｐｅａｎ Ｒｉｂｏｓｏｍａｌ ＲＮＡ ｄａｔａｂａｓｅに掲載されているプライマーであり、その他は今回の塩基配列を決定するために新たに設計したものである。尚、プライマー（１）及び（２）の末尾の（）内の数字は大腸菌における１６ＳｒＲＮＡ遺伝子上の位置を示し、Ｆはフォワード、及びＲはリバースを表す。また、プライマー（３）〜（６）の末尾の数字は、乳酸菌における１６ＳｒＲＮＡの遺伝子配列のおおよその位置を示し、Ｆはフォワード、及びＲはリバースを表す。
（１）ＡＧＡＧＴＴＴＧＡＴＣＣＴＧＧＣＴＣＡＧ（２７−Ｆ）
（２）ＣＣＡＴＴＧＴＧＧＡＡＧＡＴＴＣＣＣＴＡＣ（３５０−Ｒ）
（３）ＣＴＡＣＧＧＧＡＧＧＣＡＧＣＡＧＴＡＧＧＧＡＡＴＣ（３００−Ｆ）
（４）ＧＡＡＣＡＣＣＡＧＴＧＧＣＧＡＡＧＧＣＧ（７３０−Ｆ）
（５）ＡＴＧＴＴＧＧＧＴＴＡＡＧＴＣＣＣＧＣＡＡＣＧ（１１００−Ｆ）
（６）ＡＡＧＧＡＧＧＴＧＡＴＣＣＡＧＣＣＧＣＡ（１５４１−Ｒ）
The 16S ribosomal RNA nucleotide sequence in the lactic acid bacterium of the present invention was sequenced using the following six primers, and BLAST search was performed.
In addition, the standard strain JCM1115 of Lactobacillus buchneri was obtained from RIKEN Microbial System Preservation Project (JCM: Japan Collection of Microorganisms), and the 16S rRNA gene sequence was determined, and the isolates (SU-2, SU-4, SU-6) Each 16S rRNA gene sequence was compared. As a result, the sequences of these isolates were almost 100% identical to the standard strain, and these isolates were identified as Lactobacillus buchneri. In addition, the gene sequence of 16S rRNA of the above-mentioned standard strain was registered in NCBI Gene Bank (ACCESSION number; AB205055).
Primers used for 16S rRNA base analysis are as follows. Of the following primers, (1) and (6) are primers published in The European Ribosomal RNA database, and the others are newly designed to determine the base sequence of this time. The numbers in parentheses at the end of primers (1) and (2) indicate the position on the 16S rRNA gene in E. coli, F represents forward, and R represents reverse. The numbers at the end of the primers (3) to (6) indicate the approximate position of the gene sequence of 16S rRNA in lactic acid bacteria, F represents forward, and R represents reverse.
(1) AGAGTTTGATCCTGGCTCAG (27-F)
(2) CCATTGTGGAAGATTCCCTAC (350-R)
(3) CTACGGGAGGCAGCAGTAGGGGAATC (300-F)
(4) GAACACCAGTGGCGAAGGGCG (730-F)
(5) AGTTTGGGTTAAGTCCCCACACG (1100-F)
(6) AAGGAGGTGATCCAGCCCGCA (1541-R)
本発明における発酵組成物は、上記乳酸菌の少なくとも１種を利用して得られたことを特徴とする。
本発明における発酵組成物は、上記乳酸菌が生育している過程で生成されたものであれば良く、例えば、上記乳酸菌の培養物、該培養物から菌体を分離した培養液、該培養液の濃縮物、ペースト化物及び固形物等が挙げられる。また、乳酸菌としては生菌体、乾燥菌
等が適宜使用可能である。
以上のようにして得られる発酵組成物は食品等に好適に用いられ、ヒトが摂取する食品に加えて、非ヒトの哺乳動物用の餌、飼料などの食品にも好適に使用することができる。
The fermentation composition in the present invention is obtained using at least one of the above lactic acid bacteria.
The fermentation composition in the present invention may be any composition as long as it is produced in the process in which the lactic acid bacterium is growing. For example, a culture of the lactic acid bacterium, a culture solution obtained by separating cells from the culture, Concentrates, pastes, solids, and the like can be mentioned. Further, as the lactic acid bacteria, viable cells, dry bacteria and the like can be used as appropriate.
The fermented composition obtained as described above is suitably used for foods and the like, and can be suitably used for foods such as food and feed for non-human mammals in addition to foods taken by humans. .
本発明における漬物食品は、上記乳酸菌の少なくとも１種を添加して発酵させることにより得られることを特徴とする。
漬物食品の原料は、特に限定さないが、野菜及び海藻が好ましい。
上記野菜は、漬物に使用され得るものであれば、特に限定はされない。例えば、キュウリ、瓜、白菜、キャベツ、レタス、ほうれん草、春菊、大根、かぶ、にんじん、牛蒡、レンコン、なす、ピーマン、トマト、ふき、玉葱、セロリ、オクラ、青菜、シシトウ、唐辛子、わさび等が挙げられる。また、上記海藻としては、昆布、わかめ、アオサ、ノリ等が挙げられる。これらは一種で用いてもよく、二種以上を組み合わせて用いてもよい。
更に、上記原料以外に肉、魚等を使用することができる。これらは一種で用いてもよく、二種以上を組み合わせて用いてもよい。
The pickled food according to the present invention is obtained by adding and fermenting at least one of the above lactic acid bacteria.
The raw material of the pickled food is not particularly limited, but vegetables and seaweed are preferable.
The said vegetable will not be specifically limited if it can be used for a pickle. For example, cucumber, salmon, Chinese cabbage, cabbage, lettuce, spinach, spring chrysanthemum, radish, turnip, carrot, beef bowl, lotus root, eggplant, bell pepper, tomato, wiping, onion, celery, okra, green vegetables, shiitake, chili, wasabi It is done. Examples of the seaweed include kelp, wakame, aosa, and seaweed. These may be used alone or in combination of two or more.
Furthermore, meat, fish, etc. can be used in addition to the above raw materials. These may be used alone or in combination of two or more.
本発明における漬物食品の製造方法は、上記乳酸菌の少なくとも１種を添加して発酵させるものであれば、特に限定されない。通常の漬物製造の場合と同様に製造することができる。例えば、原料の野菜等を洗浄し、必要に応じて、アク抜き、水きり、皮の削除、適当な大きさに裁断、陰干し、及び塩等による下漬け等を行い、その後、上記乳酸菌の培養液等を添加する。その添加量は、適宜選択されるが、通常、原料１００質量％に対して、０．０１〜５０質量％であり、好ましくは０．０３〜２０質量％、より好ましくは０．０５〜１０質量％である。添加量が上記範囲内にあると、良好な食味を有する漬物食品とすることができる。
その後、暗所等にて保持する。保持温度は、通常０〜２５℃が好ましく、１０〜２０℃がより好ましい。この保持温度範囲とすることで、乳酸菌の発酵が促進され、且つ腐敗等を防ぐことできる。また、保持期間は、原料の種類及び食味の嗜好により適宜選択できるが、浅漬けの場合は１〜２日間であり、通常、３日〜１月である。この期間保持することで食味良好な漬物食品を得られる。
また、上記漬物食品はこのまま、或いは低温下で貯蔵することができる。更に、アミノ酸系調味料やソルビトールなどの甘味料で調味することもできる。
The method for producing pickled food in the present invention is not particularly limited as long as at least one of the above lactic acid bacteria is added and fermented. It can be produced in the same manner as in the case of normal pickle production. For example, washing the raw vegetables, etc., if necessary, removing acupuncture, draining, removing the skin, cutting to an appropriate size, drying in the shade, submerging with salt, etc., and then the culture solution of the lactic acid bacteria Etc. are added. The addition amount is appropriately selected, but is usually 0.01 to 50% by mass, preferably 0.03 to 20% by mass, more preferably 0.05 to 10% by mass with respect to 100% by mass of the raw material. %. When the addition amount is within the above range, a pickled food product having a good taste can be obtained.
Then hold in a dark place. The holding temperature is usually preferably 0 to 25 ° C, more preferably 10 to 20 ° C. By setting it as this holding | maintenance temperature range, fermentation of lactic acid bacteria is accelerated | stimulated and decay can be prevented. In addition, the retention period can be appropriately selected depending on the type of raw material and taste preference, but in the case of shallow pickling, it is 1 to 2 days, and usually 3 days to 1 month. By holding for this period, a pickled food with good taste can be obtained.
The pickled food can be stored as it is or at a low temperature. Furthermore, it can be seasoned with a sweetener such as an amino acid seasoning or sorbitol.
本発明における食品組成物は、上記乳酸菌の少なくとも１種を培養して得られる菌体を含むものであれば、特に限定されない。上記菌体は生菌体、乾燥菌等が適宜使用可能であり、菌体を含有する培養液等の発酵組成物でも構わない。
更に、本発明によって見出された乳酸菌は、培養して得られた培養液から遠心分離機等によって集めた乳酸菌の菌体を凍結乾燥やスプレードライヤーによって粉末にすることにより、生菌粉末素材としてもさまざまな食品へ利用できる。
The food composition in the present invention is not particularly limited as long as it contains cells obtained by culturing at least one of the above lactic acid bacteria. As the cells, live cells, dry cells and the like can be used as appropriate, and a fermentation composition such as a culture solution containing the cells may be used.
Furthermore, the lactic acid bacteria found by the present invention can be obtained as a raw bacterial powder material by lyophilizing or spraying lactic acid bacteria collected from a culture solution obtained by culturing with a centrifugal separator or the like by freeze drying or a spray dryer. Can also be used for various foods.
以下、本発明を実施例により具体的に説明する。
（１）乳酸菌の培養
鮒寿司から単離された３種類の乳酸菌（ＳＵ−２、ＳＵ−４及びＳＵ−６）とＬａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉの標準株ＪＣＭ１１１５Ｔ（以下、「標準株」と略記する。）について、上述のＭＲＳ液体培地で培養した。
培養条件は、コロニーをＭＲＳ液体培地６ｍｌに接種し、３３℃で２４時間静置培養とした。
更に、寒天培地に画線して、この寒天培地をアネロパック・ケンキ（三菱ガス化学株式会社製）と一緒にパウチへ入れて３３℃において、４８時間嫌気培養した。
尚、上記Ｌａｃｔｏｂａｃｉｌｌｕｓ ｂｕｃｈｎｅｒｉの標準株「ＪＣＭ１１１５Ｔ」は、「理化学研究所 微生物系統保存事業」から入手した。
また、ＳＵ−２、ＳＵ−４及びＳＵ−６が実施例であり、標準株が比較例である。
Hereinafter, the present invention will be specifically described by way of examples.
(1) Culture of Lactic Acid Bacteria Three types of lactic acid bacteria (SU-2, SU-4 and SU-6) isolated from sushi and standard strain JCM1115 T of Lactobacillus buchneri (hereinafter abbreviated as “standard strain”) Was cultured in the MRS liquid medium described above.
The culture conditions were such that colonies were inoculated into 6 ml of MRS liquid medium and statically cultured at 33 ° C. for 24 hours.
Further, the agar medium was streaked, and this agar medium was put into a pouch together with Aneropac Kenki (manufactured by Mitsubishi Gas Chemical Co., Ltd.) and cultured at 33 ° C. for 48 hours.
The standard strain “JCM1115 T ” of Lactobacillus buchneri was obtained from “RIKEN Microorganism System Preservation Project”.
Moreover, SU-2, SU-4, and SU-6 are examples, and a standard strain is a comparative example.
（２）乳酸菌の保存及び復元
全重量を１００重量％とした場合に、スキムミルク１０重量％及びグルタミン酸ナトリウム１重量％を含む水溶液に、寒天培地からかきとった菌体を１ｍｌあたり１０８から１０９個の菌体を含むように分散したのち、凍結乾燥する。復元は、ＭＲＳ培地に乾燥菌粉末をいれ、３３℃インキュベーター中で培養する。
または、乳酸菌の保存及び復元の別の方法として、オートクレーブしたＭＲＳ培地を１．５ｍｌづつ分注した４本の２ｍｌチューブに上記４種類の乳酸菌の培養液をそれぞれ２００μｌづつ分注し、液体窒素で凍らせてディープフリーザで保存した。
また、復元は、３３℃インキュベーターにチューブを移しておこない、さらに、解凍した培地はあらたに植え継ぎ、寒天培地に画線してコンタミの確認を行った。
(2) Storage and Restoration of Lactic Acid Bacteria When the total weight is 100% by weight, 10 8 to 10 9 per 10 ml of cells scraped from an agar medium in an aqueous solution containing 10% by weight skim milk and 1% by weight sodium glutamate. After being dispersed so as to contain individual cells, it is freeze-dried. For reconstitution, dry powder is placed in MRS medium and cultured in a 33 ° C. incubator.
Alternatively, as another method for storage and restoration of lactic acid bacteria, 200 μl of each of the above four types of lactic acid bacteria cultures is dispensed into four 2 ml tubes into which 1.5 ml of autoclaved MRS medium has been dispensed. Frozen and stored in Deep Freezer.
Restoration was performed by transferring the tube to a 33 ° C. incubator, and the thawed medium was replanted and streaked on the agar medium to check for contamination.
（３）キュウリの漬物の製造
キュウリ３本（約４４０g）を水で洗浄し、包丁で両端を切り落とし、表面の皮を１／３〜１／２程度剥く。塩をつけて板ずりし、洗う。このキュウリを約１ｃｍ幅に切り、純度９５％のアルコール２０ｍｌをふりかける。その後、刻んだキュウリに付着しているアルコール及び水をペーパータオルでふき取り、容器にあける。
次に、食塩２２ｇをふりかけ、容器内でかき混ぜる。それをビニール袋に４等分になるように分けて投入し、Ｔｗｅｅｎ８０を除いて調製した改変ＭＲＳ培地から培養した標準株、ＳＵ−２、ＳＵ−４及びＳＵ−６の培養液を各々のビニール袋に１．５ｍｌを加え、密閉した後、よく混合し、ビニール袋に密閉したまま漬物器に入れて重石を置く。
更に、冷暗所（温度１０〜２０℃）に４日間置いた後、上記キュウリの漬物について官能試験を行った。
(3) Manufacture of pickled cucumbers Three cucumbers (about 440 g) are washed with water, both ends are cut off with a knife, and the surface peel is peeled off by about 1/3 to 1/2. Add salt and wash and wash. Cut this cucumber to a width of about 1 cm and sprinkle with 20 ml of 95% pure alcohol. Thereafter, the alcohol and water adhering to the chopped cucumber are wiped off with a paper towel and poured into a container.
Next, sprinkle with 22 g of salt and stir in the container. It was put into a plastic bag divided into 4 equal parts, and the cultures of the standard strains SU-2, SU-4 and SU-6 cultured from the modified MRS medium prepared by removing Tween 80 were added to each vinyl. Add 1.5 ml to the bag, seal it, mix well, put it in a pickle container while keeping it sealed in a plastic bag, and place the weight.
Furthermore, after leaving in a cool dark place (temperature 10-20 degreeC) for 4 days, the sensory test was done about the said pickled cucumber.
（４）官能試験
官能試験は１０人のボランティアにより実施した。乳酸菌の標準株、ＳＵ−２、ＳＵ−４又はＳＵ−６の培養液をそれぞれ添加し、４日間発酵させたキュウリの漬物をそれぞれ食し、食味について４段階による評価を行った。
評価内容は、食味について、「おいしかった」、「まあおいしかった」、「酸味がある」及び「塩かどがある」から選択するものとした。官能試験の結果を以下の表３に示す。
(4) Sensory test The sensory test was carried out by 10 volunteers. A lactic acid bacteria standard strain, a culture solution of SU-2, SU-4, or SU-6 was added, and each of the pickled cucumbers fermented for 4 days was eaten, and the taste was evaluated according to four stages.
The contents of evaluation were selected from “delicious”, “delicious”, “has acidity”, and “has salted corners” for the taste. The results of the sensory test are shown in Table 3 below.
（５）白菜の漬物
白菜（２株）の外側の葉を取り除き、株のまま水で洗浄し、夜〜翌日昼まで(約１７時間)室外で干す。白菜の硬い芯を取り除き、残りを約５ｃｍのざく切りにする。４個の漬物器にざく切りにした白菜を６００ｇずつ入れ、それぞれ、塩１２g（白菜を１００重量％とした場合の重量％として２重量％）、昆布６g（白菜を１００重量％とした場合の重量％として１％）を加えてよくかき混ぜて、タカノツメ１本をのせて重石をする。冷暗所（温度１０〜２０℃）に1日置く。上記の方法で培養した標準株、ＳＵ−２、ＳＵ−４及びＳＵ−６の培養液を各々の漬物器に１ｍｌを加え、更に、冷暗所（温度１０〜２０℃）に５日間置いた後、白菜の漬物について官能試験を行った。官能試験の方法及び評価方法は、上記（３）と同様である。また、評価の結果は上記（３）の表３の結果と同じであった。
(5) Pickled Chinese cabbage Remove the leaves of Chinese cabbage (2 stocks), wash the stock with water, and dry it outdoors from night to the next day (about 17 hours). Remove the hard core of Chinese cabbage and cut the remainder into 5 cm pieces. 600g each of Chinese cabbage chopped into 4 pickles, and 12g salt (2% by weight as 100% by weight of Chinese cabbage) and 6g of kelp (100% by weight of Chinese cabbage) Add 1% as a weight%) and stir well. Place in a cool dark place (temperature 10-20 ° C) for 1 day. After adding 1 ml of the culture solution of the standard strain, SU-2, SU-4 and SU-6 cultured by the above method to each pickle container, and further placing it in a cool dark place (temperature 10-20 ° C.) for 5 days, A sensory test was conducted on the pickled Chinese cabbage. The sensory test method and the evaluation method are the same as in (3) above. Moreover, the result of evaluation was the same as the result of Table 3 in (3) above.
（６）野菜ジュースへの添加効果実験
予め煮沸殺菌した４つのガラス容器に、トマトジュース（製品名：カゴメトマトジュース カゴメ株式会社製）を各々１５０ｍｌづつ入れ、そして、上記の方法で培養した標準株、ＳＵ−２、ＳＵ−４及びＳＵ−６の各培養液１．５ｍｌを各々の容器に接種し、２５℃に３日間置いて官能試験を行った。
官能試験は、標準株、ＳＵ−２、ＳＵ−４及びＳＵ−６の培養液を接種した各トマトジュースの食味について、接種後の１日目、２日目及び３日目の３回行なった。その官能試験の結果を以下の表４に示す。
(6) Experiment on effect of addition to vegetable juice Into four glass containers previously boiled and sterilized, 150 ml each of tomato juice (product name: Kagome Tomato Juice manufactured by Kagome Co., Ltd.) was added and cultured using the above method. , SU-2, SU-4 and SU-6 were inoculated into each container and placed at 25 ° C. for 3 days for a sensory test.
The sensory test was performed three times on the first day, the second day, and the third day after inoculation for the taste of each tomato juice inoculated with the culture solution of the standard strain, SU-2, SU-4, and SU-6. . The results of the sensory test are shown in Table 4 below.
（７）実施例の効果
表３の結果より、パネラーによる本発明の乳酸菌を使用した漬物の官能評価は、実施例のＳＵ−２及びＳＵ−４は、「まあおいしかった」であり、実施例のＳＵ−６は「おいしかった」という評価であった。かかる結果より、本発明の乳酸菌を添加した試験品は、比較例の標準株による対照品に比べて優れた食味を奏することが確認できた。
また、表４の結果より、実施例のＳＵ−４及びＳＵ−６を添加したトマトジュースは、２日目にはトマト臭が低減した。また、ＳＵ−４は爽やかな酸味を呈し、優れた食味を奏する効果が確認できた。また、実施例のＳＵ−２も３日目にはトマト臭が低減し、更に、ＳＵ−４及びＳＵ−６は、３日目にはトマト臭がほとんど無くなった。かかる結果より、本発明の乳酸菌を飲食品等に添加することにより、優れた食味を奏する効果が確認できた。
(7) Effects of Examples From the results of Table 3, the sensory evaluation of pickles using the lactic acid bacteria of the present invention by the panelists was that SU-2 and SU-4 of the examples were “good” and the examples The SU-6 was rated "delicious". From these results, it was confirmed that the test product to which the lactic acid bacterium of the present invention was added had an excellent taste compared to the control product using the standard strain of the comparative example.
Moreover, from the result of Table 4, tomato juice to which SU-4 and SU-6 of Examples were added had a reduced tomato odor on the second day. Moreover, SU-4 exhibited a refreshing acidity and confirmed the effect of producing an excellent taste. In addition, the tomato odor in the SU-2 of the Example also decreased on the third day, and the tomato odor almost disappeared on the third day in the SU-4 and SU-6. From these results, the effect of producing an excellent taste could be confirmed by adding the lactic acid bacteria of the present invention to food and drink.
尚、本発明においては、上述の具体的な実施例に示すものに限らず、目的、用途に応じて本発明の範囲内で種々変更した実施例とすることができる。即ち、本発明の乳酸菌を畜肉食品、乳製品等に使用して、発酵食品を製造することができる。 In addition, in this invention, it can be set as the Example variously changed within the range of this invention according to the objective and use, without being shown to the above-mentioned specific Example. That is, fermented foods can be produced using the lactic acid bacteria of the present invention for livestock meat foods, dairy products and the like.
本発明の乳酸菌は、食品の風味と保存性を高めることから、ヒトが摂取する飲料、食品等の分野に好適に利用できる。更に、非ヒトの哺乳動物（ネコ、イヌなどのペット及び家畜動物）用の餌、飼料などの食品に好適に利用することができる。 Since the lactic acid bacteria of the present invention enhance the flavor and storage stability of foods, they can be suitably used in the fields of beverages, foods and the like taken by humans. Furthermore, it can utilize suitably for foodstuffs, such as a feed and feed for non-human mammals (pets and livestock animals such as cats and dogs).
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2008180659A JP5319186B2 (en) | 2008-07-10 | 2008-07-10 | Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2008180659A JP5319186B2 (en) | 2008-07-10 | 2008-07-10 | Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2010017132A JP2010017132A (en) | 2010-01-28 |
| JP5319186B2 true JP5319186B2 (en) | 2013-10-16 |
Family
ID=41702642
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2008180659A Active JP5319186B2 (en) | 2008-07-10 | 2008-07-10 | Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP5319186B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5175991B2 (en) * | 2011-07-04 | 2013-04-03 | 株式会社ヤサカ | Method for producing lactic acid bacteria fermented product, hypertension inhibitor, blood fat lowering agent, and fat accumulation inhibitor |
-
2008
- 2008-07-10 JP JP2008180659A patent/JP5319186B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| JP2010017132A (en) | 2010-01-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110257285B (en) | Lactobacillus plantarum with functions of efficiently degrading nitrite and strongly producing acid and application thereof | |
| Jay et al. | Modern food microbiology | |
| Kopermsub et al. | Identification of lactic acid bacteria associated with the production of plaa-som, a traditional fermented fish product of Thailand | |
| CN106754472B (en) | One plant of fruits and vegetables fermentation lactobacillus plantarum grx15 and its application | |
| JP5933650B2 (en) | Method for producing fermented food and drink | |
| CN108102959A (en) | Humanized norcholesterol lactobacillus plantarum ZY08 and its application | |
| CN109234204A (en) | A kind of pickle starter and its methods for making and using same | |
| CN105767984A (en) | Fermented chili-pepper product and preparation method thereof | |
| KR101302705B1 (en) | Method for preparing pork cutlet using capsosiphon fulvescens | |
| CN104774781B (en) | One bacillus subtilis DCU and application thereof | |
| CN113980853A (en) | Lactococcus garvieae WBT0008 capable of producing lactic acid at high yield and application thereof | |
| Muzaddadi et al. | An accelerated process for fermented fish (seedal) production in Northeast region of India | |
| CN104726359B (en) | One bacillus pumilus BP and application thereof | |
| Sharma et al. | Potential of Lactococcus lactis subsp. lactis MTCC 3041 as a biopreservative | |
| CN105533577A (en) | Method for preparing vegetable-fermented product | |
| CN104988099B (en) | A method of agent of lactic acid bacteria and bacteriocin are produced using salted vegetable waste water | |
| JP5319186B2 (en) | Lactic acid bacteria, fermented composition and pickled food obtained using the same, and food composition | |
| CN111264817A (en) | Application of direct vat set lactobacillus starter in preserved szechuan pickle pickling | |
| Belhadj et al. | Isolation, identification and antimicrobial activity of lactic acid bacteria from Algerian honeybee collected pollen | |
| JP6723427B1 (en) | Lactic Acid Bacteria with Sour Taste Inhibition and Fermented Pickle Production Method Using the Same | |
| JP2017012163A (en) | Lactobacillus plantarum llp5193 with high acid- and bile-resistance and high cell attaching ability and products containing it as the effective ingredient | |
| Zhai et al. | Evaluation of the use of malic acid decarboxylase‐deficient starter culture in NaCl‐free cucumber fermentations to reduce bloater incidence | |
| FI116191B (en) | The lactobacillus strain and its use in an acidification culture for fermenting plant material | |
| Bajwa et al. | Evaluation of probiotic properties of yeasts isolated from Sidra–an ethnic fermented fish product of north East India | |
| JP5577559B2 (en) | Novel lactic acid bacteria and method for producing fermented foods using the lactic acid bacteria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20110708 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110708 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110822 |
|
| RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20121109 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130312 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20130501 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20130613 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20130711 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 5319186 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313115 |
|
| R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |