JP5577559B2 - Novel lactic acid bacteria and method for producing fermented foods using the lactic acid bacteria - Google Patents
Novel lactic acid bacteria and method for producing fermented foods using the lactic acid bacteria Download PDFInfo
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Landscapes
- Dairy Products (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Description
本発明は、低温で優位に増殖して雑菌増殖を抑制する乳酸菌とそれを利用した発酵食品の製造方法に関する。 The present invention relates to a lactic acid bacterium that proliferates advantageously at low temperatures and suppresses the growth of various bacteria, and a method for producing a fermented food using the lactic acid bacterium.
従来、漬物を製造する際、食品に多量の食塩が添加されてきた。多量の食塩により食品の保存性を高めることができるという利点があるものの、塩分の過剰摂取が高血圧の要因となるために、現在では漬物の低塩化が進んでいる。この低塩化によって引き起こされる雑菌の増殖や、食品の品質・保存性の低下を防ぐために、さらに加熱または次亜塩素酸ナトリウム水による殺菌処理が行われている。ところが、過度の加熱処理は加熱臭の発生や、食感の損失など加工食品の品質低下を引き起こし、次亜塩素酸ナトリウム水の添加は塩素臭の発生を引き起こすという問題があった。 Conventionally, when manufacturing pickles, a large amount of salt has been added to food. Although there is an advantage that a large amount of salt can improve the storage stability of food, excessive salt intake causes high blood pressure, so that low salification of pickles is now progressing. In order to prevent the proliferation of various bacteria caused by this low salinization and the deterioration of the quality and storage stability of the food, further heat treatment or sterilization treatment with sodium hypochlorite water is performed. However, excessive heat treatment causes problems such as generation of heated odor and quality deterioration of processed foods such as loss of texture, and addition of sodium hypochlorite water causes generation of chlorine odor.
そこで、発酵食品を製造する際、地域の特産食品やその土地に由来する乳酸菌を分離し、それをスターターとして用いることで、品質の安定化、雑菌増殖の抑制、風味の向上が図られてきた。例えば、白神山地の腐葉土から分離された乳酸菌を用いた発酵食品の製造方法（特許文献１）や、発酵キムチから分離される乳酸菌ラクトバチルス・サケＨＳ１を用いた漬物の製造方法（特許文献２）などが知られている。しかし、一般の雑菌が増殖しない低温下で優位に増殖し、雑菌の増殖抑制と発酵食品の食味の向上に寄与する乳酸菌は稀少である。 Therefore, when producing fermented foods, the local specialty foods and lactic acid bacteria derived from the local area are separated and used as a starter to stabilize the quality, suppress the growth of bacteria, and improve the flavor. . For example, a method for producing fermented foods using lactic acid bacteria isolated from humus in Shirakami Mountains (Patent Document 1) and a method for producing pickles using lactic acid bacteria Lactobacillus salmon HS1 separated from fermented kimchi (Patent Document 2) Etc. are known. However, lactic acid bacteria that predominately grow at low temperatures at which general bacteria do not grow and contribute to the suppression of the growth of bacteria and the taste of fermented foods are rare.
そこで、本発明は上記問題点に鑑み、低食塩および低温下で保存する加工原料野菜等の保存性、食味を向上させるために、低温で速やかに増殖し雑菌増殖を抑制する乳酸菌およびそれを使用した雑菌増殖の抑制方法または発酵食品の製造方法を提供することを課題とする。 Therefore, in view of the above problems, the present invention uses a lactic acid bacterium that rapidly grows at a low temperature and suppresses the growth of miscellaneous bacteria in order to improve the storage stability and taste of processed raw vegetables and the like that are stored under low salt and low temperature. It is an object of the present invention to provide a method for inhibiting the growth of miscellaneous bacteria or a method for producing a fermented food.
上記課題を達成するため鋭意検討した結果、低温で優位に増殖して雑菌増殖を抑制する新規の乳酸菌を見出し、その乳酸菌を利用した発酵食品の製造方法を開発した。具体的には、前記乳酸菌は、新潟県魚沼地域の雪室で保存した野沢菜漬けから分離された３種類の低温増殖性乳酸菌である。また、これら３種類の乳酸菌を用いた雑菌増殖の抑制方法または発酵食品の製造方法である。 As a result of intensive studies to achieve the above-mentioned problems, a novel lactic acid bacterium that proliferates advantageously at low temperatures and suppresses the growth of various bacteria was found, and a method for producing fermented foods using the lactic acid bacterium was developed. Specifically, the lactic acid bacteria are three types of cold-growing lactic acid bacteria isolated from Nozawana pickles preserved in a snow room in the Uonuma area of Niigata Prefecture. Further, it is a method for suppressing the growth of miscellaneous bacteria using these three types of lactic acid bacteria or a method for producing fermented foods.
すなわち、本発明の乳酸菌は、ラクトバチルス サケイ ＵＯＮＵＭＡ−１（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−１：受託番号ＮＩＴＥ Ｐ−１３２３）、ラクトバチルス サケイ ＵＯＮＵＭＡ−２（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−２：受託番号ＮＩＴＥ Ｐ−１３２４）またはラクトバチルス サケイ ＵＯＮＵＭＡ−３（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−３：受託番号ＮＩＴＥ Ｐ−１３２５）であることを特徴とする。 That is, the lactic acid bacteria of the present invention include Lactobacillus sake UONUMA-1 (Lactobacillus sakei UONUMA-1: accession number NITE P-1323) and Lactobacillus sake UONUMA-2 (Lactobacillus sakei UONUMA-2: accession number NITE P-13). It is characterized by being Lactobacillus sake UONUMA-3 (Lactobacillus sake UONUMA-3: accession number NITE P-1325).
また、本発明の発酵食品の製造方法は、請求項１に記載のいずれかの乳酸菌を用いて、発酵させるステップを含むことを特徴とする。 Moreover, the manufacturing method of the fermented food of this invention is characterized by including the step fermented using the lactic acid bacteria in any one of Claim 1.
また、前記発酵食品の製造方法は、請求項１に記載のすべての乳酸菌を混合して用い、発酵させるステップを含むことを特徴とする。 In addition, the method for producing fermented food includes a step of mixing and using all the lactic acid bacteria according to claim 1 and fermenting.
また、前記発酵食品の製造方法は、４〜６℃の低温において発酵させることを特徴とする。 Moreover, the manufacturing method of the said fermented food is characterized by making it ferment at the low temperature of 4-6 degreeC.
さらに、本発明の発酵食品は、請求項２ないし４のいずれか１つに記載の製造方法により得られることを特徴とする。 Furthermore, the fermented food of this invention is obtained by the manufacturing method as described in any one of Claim 2 thru | or 4.
本発明によれば、乳酸菌が低温（４℃）でも優位に増殖するため、この乳酸菌を漬物等のスターターとして低温で利用することにより異味異臭の原因となる雑菌の増殖を抑制し、発酵食品の品質劣化を抑えることができる。 According to the present invention, since lactic acid bacteria proliferate predominantly even at low temperatures (4 ° C.), the use of the lactic acid bacteria as a starter for pickles etc. at low temperatures suppresses the growth of miscellaneous bacteria that cause off-flavor odors. Quality degradation can be suppressed.
本発明の乳酸菌ラクトバチルス サケイ ＵＯＮＵＭＡ−１（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−１）、ラクトバチルス サケイ ＵＯＮＵＭＡ−２（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−２）、ラクトバチルス サケイ ＵＯＮＵＭＡ−３（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−３）は、新潟県魚沼地域の雪室で保存した株式会社大沢加工製の野沢菜漬けから分離されたラクトバチルス サケイ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ）に属する新規の菌であり、独立行政法人製品評価技術基盤機構特許微生物寄託センター（千葉県木更津市かずさ鎌足２−５−８）に、２０１２年４月１８日付で、それぞれ受託番号ＮＩＴＥ Ｐ−１３２３、受託番号ＮＩＴＥ Ｐ−１３２４、受託番号ＮＩＴＥ Ｐ−１３２５として寄託されている。 Lactic acid bacteria Lactobacillus salmon UONUMA-1 (Lactobacillus sakei UONUMA-1), Lactobacillus salmon UONUMA-2 (Lactobacillus sakei UONUMA-2), Lactobacillus salmon UONUMA-3 (Lactobacillus Numa) It is a new fungus belonging to Lactobacillus sakei isolated from Nozawana pickles made by Osawa Co., Ltd., preserved in a local snow room, and is a patent microorganism deposit center of the National Institute of Technology and Evaluation (CHIBA) In Kisarazu City Kazusa Kamashikaga 2-5-8) on April 18, 2012, accession number NITE P-1323 and accession number NITE P-1324, respectively. It has been deposited as accession number NITE P-1325.
また、本発明の発酵食品の製造方法は、本発明の乳酸菌ラクトバチルス サケイ ＵＯＮＵＭＡ−１、ラクトバチルス サケイ ＵＯＮＵＭＡ−２、ラクトバチルス サケイ ＵＯＮＵＭＡ−３をそれぞれ単独で用いた場合と比較して、３種類を混合して用いた場合のほうが、得られた発酵食品の官能評価が高かったため、３種類の前記乳酸菌を混合して用いることが好ましい。 Moreover, the manufacturing method of the fermented food of this invention has three types compared with the case where the lactic acid bacteria Lactobacillus salmon UONUMA-1, Lactobacillus salmon UONUMA-2, and Lactobacillus salmon UONUMA-3 of this invention are each used independently. Since the sensory evaluation of the obtained fermented food was higher when the mixture was used, it is preferable to mix and use the three types of lactic acid bacteria.
また、本発明の乳酸菌は４℃でも優位に増殖することが確認されたことから、本発明の発酵食品は４〜６℃の低温で発酵させることが好ましい。これにより、本発明の乳酸菌を漬物等のスターターとして低温で利用することにより異味異臭の原因となる雑菌の増殖を抑制し、発酵食品の品質劣化を抑えることができる。 Moreover, since it was confirmed that the lactic acid bacteria of this invention proliferate preferentially also at 4 degreeC, it is preferable to ferment the fermented food of this invention at the low temperature of 4-6 degreeC. Thereby, by using the lactic acid bacteria of the present invention as a starter such as pickles at a low temperature, it is possible to suppress the growth of miscellaneous bacteria that cause off-flavors and to suppress the quality deterioration of the fermented food.
したがって、本発明の乳酸菌は発酵食品の製造に利用でき、該発酵食品としては、特定のものに限定されないが、野沢菜漬けやキムチなどの漬物、またはヨーグルト様食品が好適である。 Therefore, the lactic acid bacteria of the present invention can be used for the production of fermented foods, and the fermented foods are not limited to specific ones, but pickles such as Nozawana pickles and kimchi, or yogurt-like foods are suitable.
なお、本発明は上記実施形態に限定されるものではなく、本発明の思想を逸脱しない範囲で種々の変形実施が可能である。 The present invention is not limited to the above embodiment, and various modifications can be made without departing from the spirit of the present invention.
本発明の新規乳酸菌の分離および同定について説明する。 The separation and identification of the novel lactic acid bacteria of the present invention will be described.
雪室に４週間保存した野沢菜漬けの漬液を野沢菜漬けとともにストマッカーで均質にした。その一部を乳酸菌分離用のＭＲＳ白亜寒天培地で培養し、培地に添加した炭酸カルシウムを溶解してクリアゾーン（透明帯）を形成した乳酸菌８０株を採取した。これらを個々にＭＲＳ培地に接種し培養後、ゲノムＤＮＡ多型性を利用したランダム増幅多型ＤＮＡ（ＲＡＰＤ）分析により各株のグループ分けを行い、７つのグループに分類した。次に各グループから１株ずつ選抜し、１６ＳリボソームＤＮＡの部分塩基配列の相同性解析を行った結果、全てラクトバチルス サケイ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ）と同定した。 The pickled solution of Nozawana pickled in the snow room for 4 weeks was homogenized with a stomacher along with Nozawana pickled. A part thereof was cultured on an MRS chalk agar medium for separating lactic acid bacteria, and 80 strains of lactic acid bacteria that formed a clear zone by dissolving calcium carbonate added to the medium were collected. These were individually inoculated into MRS medium and cultured, and then each strain was grouped by random amplified polymorphic DNA (RAPD) analysis using genomic DNA polymorphism and classified into 7 groups. Next, one strain was selected from each group, and as a result of homology analysis of the partial base sequence of 16S ribosomal DNA, all were identified as Lactobacillus sakei.
次に、分離した８０株の乳酸菌株のうち、多くの菌株（６３株）が属する３グループから１株ずつ選抜し、これら乳酸菌が産生する乳酸の旋光性を分析した。その結果、酢酸ナトリウムを含まない培地で培養すると産生する乳酸はＤＬ型を示し、培地に酢酸ナトリウムを含むとＬ型を示した。これは、ラクトバチルス サケイ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ）に属する乳酸菌が持つ特異性であり、このことからも本選抜菌株はラクトバチルス サケイ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ）と同定した。 Next, among the 80 isolated lactic acid strains, one strain was selected from 3 groups to which many strains (63 strains) belong, and the optical rotation of lactic acid produced by these lactic acid bacteria was analyzed. As a result, lactic acid produced when cultured in a medium not containing sodium acetate showed DL type, and when the medium contained sodium acetate, it showed L type. This is the specificity of lactic acid bacteria belonging to Lactobacillus sakei, and from this fact, this selected strain was identified as Lactobacillus sakei.
次に、選抜した３菌株の糖類の資化性を検討した。３菌株全てがマルトースを資化せず、アミグダリン、メリビオース、β−ゲンチオビオースの資化性は各菌株間で異なった。 Next, the assimilation property of saccharides of the selected three strains was examined. All three strains did not assimilate maltose, and the assimilation properties of amygdalin, melibiose, and β-gentiobiose were different among the strains.
いずれの菌株も既知乳酸菌株（ラクトバチルス サケイ標準株 ＮＢＲＣ３５４１、ＪＣＭ１１５７）と異なる資化性を示したことから新規な菌株と認めた。本発明の３種類のラクトバチルス サケイ菌株は、ラクトバチルス サケイ ＵＯＮＵＭＡ−１（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−１）、ラクトバチルス サケイ ＵＯＮＵＭＡ−２（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−２）、ラクトバチルス サケイ ＵＯＮＵＭＡ−３（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｓａｋｅｉ ＵＯＮＵＭＡ−３）と命名され、独立行政法人製品評価技術基盤機構特許微生物寄託センターに、それぞれ受託番号ＮＩＴＥ Ｐ−１３２３、受託番号ＮＩＴＥ Ｐ−１３２４、受託番号ＮＩＴＥ Ｐ−１３２５として寄託されている。なお、以下において、本発明の３種類のラクトバチルス サケイ菌株をそれぞれＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３という。 All strains were recognized as novel strains because they showed different assimilability from known lactic acid strains (Lactobacillus sakei standard strains NBRC3541, JCM1157). The three types of Lactobacillus salmon strains of the present invention are Lactobacillus salmon UONUMA-1 (Lactobacillus sakei UONUMA-1), Lactobacillus salmon UONUMA-2 (Lactobacillus sakei UONUMA-2), -3) and deposited at the National Institute of Technology and Evaluation Patent Microorganism Deposit Center under the accession numbers NITE P-1323, NITE P-1324, and NITE P-1325, respectively. In the following, the three types of Lactobacillus sakei strains of the present invention are referred to as UONUMA-1, UONUMA-2, and UONUMA-3, respectively.
本発明の乳酸菌と、その他乳酸菌の生育に与える培養温度の影響について説明する。 The influence of the culture temperature on the growth of the lactic acid bacteria of the present invention and other lactic acid bacteria will be described.
本発明の乳酸菌（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）と、既知乳酸菌（ＮＢＲＣ３５４１、ＪＣＭ１１４９）をＭＲＳ培地にそれぞれ接種し、培養温度５、１０、１５℃で静置培養して増殖量を吸光度６６０ｎｍで測定した。本発明の乳酸菌（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）は、いずれの温度でも速やかに増殖し、特に５℃においても良好な生育を示した（図１）。このことは一般の雑菌が増殖できない低温で発酵食品が製造できることを示唆する。 Lactic acid bacteria of the present invention (UONUMA-1, UONUMA-2, UONUMA-3) and known lactic acid bacteria (NBRC3541 and JCM1149) are inoculated into MRS medium, respectively, and cultured at 5, 10 and 15 ° C., and cultured to stand for growth. Was measured at an absorbance of 660 nm. The lactic acid bacteria of the present invention (UONUMA-1, UONUMA-2, UONUMA-3) proliferated rapidly at any temperature, and showed good growth especially at 5 ° C. (FIG. 1). This suggests that fermented foods can be produced at low temperatures where general bacteria cannot grow.
本発明の乳酸菌を用いた糠床の製造方法について説明する。 A method for producing a cocoon bed using the lactic acid bacteria of the present invention will be described.
米糠４００ｇに１００℃に加熱した食塩水（食塩５％ｗ／ｖ）６００ｇを加え、よく混合、冷却して糠床を作製した。本発明のラクトバチルス サケイ（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）と既知乳酸菌（ＮＢＲＣ３５４１、ＪＣＭ１１４９）の懸濁液をそれぞれ５ｍＬ糠床に接種し４℃で発酵させた。なお、乳酸菌の接種量は、ＭＲＳ培地で３０℃、２４時間培養し集菌して滅菌水で洗浄した後、吸光度（６６０ｎｍ）が１になるように調整した。所定の期間発酵させた後、一部を取り出し乳酸菌数を計測した（図２）。また、米糠１０ｇを１００ｍＬ滅菌生理食塩水に懸濁し、さらに１００倍希釈してシャーレ上の標準寒天培地に１ｍＬ接種、混釈した。これを３０℃で２日間培養して、現れた雑菌類を図３に示した。 To 400 g of rice bran, 600 g of brine (salt 5% w / v) heated to 100 ° C. was added, mixed well, and cooled to prepare a straw bed. The suspensions of Lactobacillus salmoni (UONUMA-1, UONUMA-2, UONUMA-3) and known lactic acid bacteria (NBRC3541, JCM1149) of the present invention were each inoculated into a 5 mL bed and fermented at 4 ° C. The amount of lactic acid bacteria inoculated was adjusted so that the absorbance (660 nm) would be 1 after culturing in MRS medium at 30 ° C. for 24 hours, collecting and washing with sterilized water. After fermentation for a predetermined period, a part was taken out and the number of lactic acid bacteria was counted (FIG. 2). Further, 10 g of rice bran was suspended in 100 mL of sterilized physiological saline, further diluted 100 times, and inoculated and mixed with 1 mL of a standard agar medium on a petri dish. This was cultured at 30 ° C. for 2 days, and the germs that appeared were shown in FIG.
本発明の乳酸菌（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）を接種した糠床は速やかに乳酸菌数が増加、１２〜１７日目には１０９オーダーに達し、その後高い菌数を維持した。また、乳酸菌数の増加とともに標準寒天培地上に現れるエンテロバクター属などの腸内細菌科に属する細菌類が減少した。このことから、本発明の乳酸菌は、低温で速やかに増殖し、雑菌の増殖を抑制することが明らかとなった。 Nukayuka inoculated with lactic acid bacteria (UONUMA-1, UONUMA-2 , UONUMA-3) of the present invention is rapidly increased lactobacilli count, the 12 to 17 day reached 10 9 order was maintained thereafter high number of bacteria . In addition, the number of bacteria belonging to the family Enterobacteriaceae such as Enterobacter, which appears on the standard agar medium, decreased with the increase in the number of lactic acid bacteria. From this, it became clear that the lactic acid bacteria of the present invention rapidly grow at low temperatures and suppress the growth of miscellaneous bacteria.
本発明の乳酸菌を用いた発酵野沢菜の製造方法について説明する。 A method for producing fermented Nozawana using the lactic acid bacteria of the present invention will be described.
下漬けした野沢菜（塩分約５％）２．５ｋｇをポリエチレン袋（０．０５×８００×９００ｍｍ）に入れ、その袋を５Ｌ容ポリエチレン製漬物容器に入れた。そこに調味液（醤油、食塩、還元水飴、調味料（アミノ酸等）、酸味料を含む）１．２Ｌを添加し、袋上部の空気を抜いて、輪ゴムで止めた。一方、本発明の乳酸菌（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）の懸濁液をそれぞれＭＲＳ培地１０ｍＬに接種した。３０℃で２４時間培養し、集菌して、滅菌水で洗浄した。再度、滅菌水に懸濁して混合し１０ｍＬとし、これを乳酸菌懸濁液とした。乳酸菌懸濁液を上記野沢菜に接種し、４℃で１０日間保存後、さらに０℃で１０日間保存した。 2.5 kg of Nozawana (salt content of about 5%) soaked in a container was placed in a polyethylene bag (0.05 × 800 × 900 mm), and the bag was placed in a 5 L polyethylene pickle container. 1.2 L of seasoning liquid (including soy sauce, salt, reduced starch syrup, seasoning (including amino acids, etc.) and acidulant) was added, and the air at the top of the bag was evacuated and stopped with a rubber band. On the other hand, suspensions of the lactic acid bacteria of the present invention (UONUMA-1, UONUMA-2, UONUMA-3) were each inoculated into 10 mL of MRS medium. The cells were cultured at 30 ° C. for 24 hours, collected, and washed with sterilized water. Again, it was suspended in sterilized water and mixed to make 10 mL, which was used as a lactic acid bacteria suspension. The lactic acid bacteria suspension was inoculated into Nozawana and stored at 4 ° C. for 10 days, and further stored at 0 ° C. for 10 days.
試作した発酵野沢菜の性状と官能評価の結果を表２に示した。非接種区は保存開始時と比較してｐＨ、一般細菌数、乳酸菌数のいずれも大きな変化はなく、市販品と同等の食味であった。一方、乳酸菌接種区では乳酸菌の増加に伴ってｐＨが低下したが、パネル６名による官能評価は適度な酸味が好評であった。 Table 2 shows the properties and sensory evaluation results of the prototype Nozawana. In the non-inoculated area, the pH, the number of general bacteria, and the number of lactic acid bacteria were not significantly changed compared to the time when the storage was started, and the taste was the same as that of the commercially available product. On the other hand, in the lactic acid bacteria inoculation zone, the pH decreased as the number of lactic acid bacteria increased, but the sensory evaluation by the six panelists was popular for its moderate acidity.
本発明の乳酸菌を用いたキムチの製造方法について説明する。 A method for producing kimchi using the lactic acid bacteria of the present invention will be described.
白菜４株（約８ｋｇ）を４つ割りし、２５Ｌ容ポリエチレン製漬物容器に入れた。そこに８％（ｗ／ｖ）食塩水８Ｌを添加した。次に押蓋をして重石（１２ｋｇ）を載せ、冷蔵庫（０℃）で一晩漬けた。一方、乳酸菌は、本発明の乳酸菌（ＵＯＮＵＭＡ−１、ＵＯＮＵＭＡ−２、ＵＯＮＵＭＡ−３）と、既知乳酸菌（ＮＢＲＣ３５４１、ＪＣＭ１１４９、特許文献２（特許第３０９１１９６号）の乳酸菌））の懸濁液をそれぞれＭＲＳ培地５０ｍＬに接種した。３０℃で２４時間培養し、集菌して、滅菌水で洗浄した後、再度、滅菌水１０ｍＬに懸濁し、乳酸菌懸濁液とした。 Four Chinese cabbages (about 8 kg) were divided into four and placed in a 25 L polyethylene pickle container. 8% (w / v) saline 8 L was added thereto. Next, it was put on a lid, placed on heavy stone (12 kg), and soaked overnight in a refrigerator (0 ° C.). On the other hand, lactic acid bacteria are suspensions of lactic acid bacteria of the present invention (UONUMA-1, UONUMA-2, UONUMA-3) and known lactic acid bacteria (NBRC3541, JCM1149, lactic acid bacteria of Patent Document 2 (Patent No. 3091196)), respectively. 50 mL of MRS medium was inoculated. The cells were cultured at 30 ° C. for 24 hours, collected, washed with sterilized water, and then suspended again in 10 mL of sterilized water to obtain a lactic acid bacteria suspension.
翌日、漬物容器から白菜を取り出し、ザルにあけ、重石を載せて、余分な水分を除いた。このとき、白菜重量は約６ｋｇであった。下漬けした白菜を１ｋｇずつ３Ｌ容プラスチック製漬物容器に分取し、乳酸菌懸濁液を１０ｍＬ接種した。調味液を約１６５ｇ添加して下漬け白菜とよく混合し、押蓋をした。５℃で１０日間保存した。なお、上記調味料は以下のように調製した。市販キムチ用調味料（原材料：ニンニク、蜜柑、林檎、生姜、食塩、砂糖・ブドウ糖果糖液糖、魚介エキス（カツオ、イカ）、唐辛子、醸造酢、昆布、調味料（アミノ酸等）、酸味料、安定剤（キサンタン）、パプリカ色素、食塩濃度１４％）９００ｇ、イカ塩辛（食塩濃度１７％）９０ｇ、砂糖７１ｇをミキサーで攪拌した。 The next day, the Chinese cabbage was taken out of the pickle container, opened in a colander, placed on a heavy stone to remove excess water. At this time, the Chinese cabbage weight was about 6 kg. 1 kg each of the pickled Chinese cabbage was dispensed into a 3 L plastic pickle container, and 10 mL of the lactic acid bacteria suspension was inoculated. About 165 g of seasoning liquid was added and mixed well with the pickled Chinese cabbage and covered with a lid. Stored at 5 ° C. for 10 days. In addition, the said seasoning was prepared as follows. Seasoning for Kimchi (Ingredients: Garlic, Tangerine, Apple, Ginger, Salt, Sugar / Glucose fructose liquid sugar, Seafood extract (bonito, Squid), Pepper, Brewed vinegar, Kelp, Seasoning (amino acids, etc.), Acidulant, Stabilizer (xanthan), paprika pigment, salt concentration 14%) 900 g, squid salty salt (salt concentration 17%) 90 g, and sugar 71 g were stirred with a mixer.
試作したキムチの性状と官能評価の結果を表３に示した。乳酸菌数は、乳酸菌接種前では２．１×１０２個／ｇであったが、乳酸菌を接種して５℃１０日間保存した後ではいずれの区も１０８個／ｇ以上となった。ｐＨは、ＮＢＲＣ３５４１株接種区では４．７０とやや高く、その他の区では４．４３〜４．５６と大差はなかった。官能評価はパネル１５名により行い、特許文献２の乳酸菌を接種した区を基準（３点）とし、「好ましい」を５点、「やや好ましい」を４点、「やや好ましくない」を２点、「好ましくない」を１点として採点した。本発明の乳酸菌は評価が高く、特にＵＯＮＵＭＡ−２株接種区及び３種類の乳酸菌を混合した区ではパネルの平均点が３．３で、特許文献２の乳酸菌の評価を上回った。 Table 3 shows the properties and sensory evaluation results of the experimental kimchi. The number of lactic acid bacteria was 2.1 × 10 2 cells / g before inoculation with lactic acid bacteria, but after inoculation with lactic acid bacteria and storage at 5 ° C. for 10 days, all the groups were 10 8 cells / g or more. The pH was slightly high at 4.70 in the NBRC3541 inoculation group, and was not significantly different from 4.43 to 4.56 in the other groups. The sensory evaluation was performed by 15 panelists, and the group (3 points) inoculated with the lactic acid bacteria of Patent Document 2 was used as the standard (3 points), “preferably” 5 points, “slightly preferable” 4 points, “slightly unfavorable” 2 points, "Unfavorable" was scored as one point. The lactic acid bacteria of the present invention were highly evaluated, and the average score of the panel was 3.3 particularly in the group in which the UONUMA-2 strain inoculated group and three types of lactic acid bacteria were mixed, exceeding the evaluation of the lactic acid bacteria in Patent Document 2.
以上のように本発明は、漬物製造、農産加工および発酵食品製造に利用できる。 As described above, the present invention can be used for pickle production, agricultural processing, and fermented food production.
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