KR20080065667A - Lactic acid bacterium having immunoregulatory activity derived from moromi for wine fermentation - Google Patents

Lactic acid bacterium having immunoregulatory activity derived from moromi for wine fermentation Download PDF

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KR20080065667A
KR20080065667A KR1020087012297A KR20087012297A KR20080065667A KR 20080065667 A KR20080065667 A KR 20080065667A KR 1020087012297 A KR1020087012297 A KR 1020087012297A KR 20087012297 A KR20087012297 A KR 20087012297A KR 20080065667 A KR20080065667 A KR 20080065667A
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다카코 야마무라
다에코 이이노
교코 아시카리
게이이치 아베
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Abstract

The invention relates to a bacterium belonging to the genus Lactobacillus having an immunoregulatory activity, a composition containing the same, and a method of efficiently isolating a lactic acid bacterium having an immunoregulatory activity. The bacterium belonging to the genus Lactobacillus of the invention has an immunoregulatory activity, and if the composition of the invention is taken or the like, a decrease in immune function can be suppressed by activating the immune function. Further, according to the invention, a lactic acid bacterium having an immunoregulatory activity can be efficiently obtained.

Description

와인 발효 모로미 유래의 면역 조절 작용을 갖는 유산균{LACTIC ACID BACTERIUM HAVING IMMUNOREGULATORY ACTIVITY DERIVED FROM MOROMI FOR WINE FERMENTATION}LACTIC ACID BACTERIUM HAVING IMMUNOREGULATORY ACTIVITY DERIVED FROM MOROMI FOR WINE FERMENTATION

본 발명은, 면역 조절 작용을 갖는 락토바실러스속균, 그것을 함유하는 조성물, 및 면역 조절 작용을 갖는 유산균의 효율이 양호한 분리 방법에 관한 것이다.This invention relates to the Lactobacillus genus which has an immunomodulatory effect, the composition containing it, and the isolation | separation method with good efficiency of the lactic acid bacterium which has an immunomodulatory effect.

유산균은 정장 작용이나 면역 부활 작용과 같은 유익한 생리 활성을 갖는 것으로 알려져 있다. 또, 일반 소비자에게 건강 효능감이 있는 소재로서의 인지도도 높다. 이와 같은 유익한 생리 활성을 갖는 유산균의 대부분은 발효 유제품이나 장관 (腸管) 으로부터 분리된 동물 소재 유래의 유산균이다.Lactic acid bacteria are known to have beneficial physiological activities such as intestinal action and immunostimulating action. Moreover, the recognition as a material with health efficacy for a general consumer is also high. Most of the lactic acid bacteria having such beneficial physiological activity are lactic acid bacteria derived from animal material separated from fermented dairy products or intestinal tract.

한편으로, 식물 소재를 분리원으로 하는 식물성 유산균에 있어서도 면역 부활 작용을 갖는 주가 발견되고 있다. 예를 들어, 일본 공개특허공보 평10-167972호 (공개일 : 1998년 6월 23일) 에는 면역 부활 작용을 갖는 유산균으로서, 락토바실러스·플란타룸 (Lactobacillus plantarum) L-137 주가 기재되어 있다. 또, 쿄토의 전통적인 채소 절임인「스구키」로부터 분리된 락토바실러스·브레비스 (Lactobacillus brevis) Labre 주나,「시바츠케」로부터 분리된 락토바실러스·펜토수스 (Lactobacillus pentosus) DA74N 주 등도 면역 부활 작용을 갖는 것으로 알 려져 있다 (참조 : (1) 일본 특허공보 평7-55908호 (공개일 : 1994년 7월 26일), (2) Pasken Journal 15. 21-26, 2002, 키시 아츠코, 코쿠보 아오이, 아카타니 카오루, 오기야 에리코, 후지타 테츠야, 키시다 모타로 : 유용 유산균 선정을 위한 스크리닝 1) 사람 말초혈에 있어서의 식물성 유산균의 시험관 내 (in vitro) 면역 부활 효과, (3) 제 6 회 장내 세균학회 (2002.05.30-31) 요지, 아카타니 카오루, 키시 아츠코, 오기야 에리코, 코쿠보 아오이, 후지타 테츠야, 키시다 모타로).On the other hand, the strain which has an immune activating effect is discovered also in the vegetable lactic acid bacteria which uses a plant material as a separation source. For example, Japanese Laid-Open Patent Publication No. 10-167972 (published: June 23, 1998) discloses Lactobacillus plantarum L-137 strain as a lactic acid bacterium having an immunostimulating action. . In addition, Lactobacillus brevis Labre strain isolated from Kyoto's traditional pickles Suguki, and Lactobacillus pentosus DA74N strain isolated from Shibatsuke also have immunosuppressive action. (Reference: (1) Japanese Patent Publication No. 7-55908 (published: July 26, 1994), (2) Pasken Journal 15. 21-26, 2002, Kishi Atsuko, Kokubo Aoi, Akatani Kaoru, Eriko Ogiya, Tetsuya Fujita, Mota Kishida: Screening for Selection of Useful Lactic Acid Bacteria 1) In Vitro Immune Revitalization Effect of Vegetable Lactic Acid Bacteria in Human Peripheral Blood, (3) 6th Intestine Bacterial Society (2002.05.30-31) Yoji, Akatani Kaoru, Kishi Atsuko, Eriko Ogiya, Kokubo Aoi, Fujita Tetsuya, and Kishida Motaro).

식물 소재를 분리원으로 하는 식물성 유산균은, 채소 절임이나 주류의 양조에 깊게 관련되어 있는 것으로 알려져 있고, 양조 공정으로부터는 수많은 균을 분리할 수 있다. 상기 이외에도 면역 부활 작용 등의 유용한 생리 활성을 갖는 균주가 존재하는 것으로 생각된다. 예를 들어, 주류의 양조 공정 중 와인 양조 공정으로부터 분리되는 균주로, 분리 빈도가 높은 주로 락토바실러스·플란타룸을 들 수 있다. 그러나, 이와 같은 주류의 양조 공정으로부터 분리된 균에 대해서는, 양조에 있어서의 작용이나 향미 등에 대한 기여에 관한 연구가 주요하고, 균 자체가 생체에 미치는 효능에 대해서는 그다지 연구되고 있지 않다. 그러므로, 주류 양조 공정으로부터 분리되는 유산균 중에서 유용한 생리 활성을 갖는 균주를 발견하는 것을 기대할 수 있다. 또, 식물성 유산균을 사용하면 독특한 발효취를 발생하는 경우가 있어, 제조하는 식품의 종류에 따라서는, 관능에 대한 대책, 배려를 필요로 하는 경우가 있다.Vegetable lactic acid bacteria which use plant material as a separation source are known to be deeply involved in pickling vegetables and brewing of alcoholic beverages, and many bacteria can be separated from the brewing process. In addition to the above, it is considered that there are strains having useful physiological activities such as immunostimulating action. For example, as a strain isolate | separated from the wine brewing process of the liquor brewing process, the mainly Lactobacillus plantarum with high isolation | separation frequency is mentioned. However, for the bacteria isolated from the liquor brewing process of such liquor, research on contribution to the action, flavor, etc. in the brewing is important, and the effect of the bacteria on the living body is not studied very much. Therefore, it can be expected to find strains with useful physiological activity among the lactic acid bacteria isolated from the mainstream brewing process. In addition, when the lactic acid bacteria are used, a unique fermentation odor may be generated, and depending on the type of food to be produced, there may be a need for measures and considerations for the sensory function.

발명의 개시Disclosure of the Invention

본 발명은, 상기의 현상황을 감안하여, 면역 조절 작용을 갖는 유산균, 그것을 함유하는 조성물, 및 면역 조절 작용을 갖는 유산균을 분리하는 방법을 제공하는 것을 목적으로 한다.It is an object of the present invention to provide a method for separating lactic acid bacteria having an immunomodulatory action, a composition containing the same, and a lactic acid bacteria having an immunomodulatory action in view of the present situation.

본 발명자들은 이러한 과제를 해결하기 위해, 주류 양조 공정에서의 생성품으로부터 분리된 유산균에 대해, 그 면역 조절 작용을 조사하고, 그 중에서 선택한 락토바실러스·플란타룸 및 락토바실러스·브레비스에 속하는 유산균에 대해, 보다 상세하게 면역 조절 작용을 조사하였다. 그 결과, 당해 유산균은 높은 면역 조절 작용을 갖고, 또 그것을 사용한 발효물은 양호한 향미를 나타내는 것을 알아내어, 본 발명을 완성시키기에 이르렀다.MEANS TO SOLVE THE PROBLEM In order to solve such a subject, the present inventors investigated the immunomodulatory action with respect to the lactic acid bacteria isolate | separated from the product in a liquor brewing process, and to the lactic acid bacteria which belong to the selected Lactobacillus plantarum and Lactobacillus brevis among them. In this regard, the immunomodulatory action was examined in more detail. As a result, the lactic acid bacterium had a high immunomodulatory effect, and it was found that the fermented product using the same exhibited good flavor, thus completing the present invention.

구체적으로는, 주류 양조 공정에서의 생성품으로부터 분리된 유산균에 대해, Th1 형 사이토카인의 인터류킨 12 (이하「IL-12」라고 약기한다) 유도능을 비교한 결과, 고효율로 면역 조절 작용을 갖는 유산균을 분리할 수 있는 것을 알아내고, 특히 락토바실러스·플란타룸, 및 락토바실러스·브레비스에 속하는 균주가, 마우스로부터 조제한 복강 매크로파지에 대해 인 비트로로 처리한 경우에 IL-12 의 산생을 상승시키는 것을 알아냈다. 또한, 마우스 복강 내에 대한 인 비보에서의 가열 균액 투여를 실시하여, Th1 형 사이토카인의 IL-12 의 산생을 유도하는 것도 확인하였다.Specifically, the lactic acid bacteria isolated from the product in the mainstream brewing process were compared with the induction ability of Th1 cytokine to interleukin 12 (hereinafter abbreviated as "IL-12"). It has been found that lactic acid bacteria can be isolated, and in particular, strains belonging to Lactobacillus plantarum and Lactobacillus brevis increase the production of IL-12 when treated in vitro to celiac macrophages prepared from mice. I found out. In addition, it was also confirmed that heating bacterial solution was administered in vivo to the mouse intraperitoneal to induce production of IL-12 of Th1-type cytokine.

즉, 본 발명은, 주류 제조 공정의 미생물 함유 성분에 포함되는, 면역 조절 작용을 갖는 락토바실러스속균이다. 특히, 미생물 함유 성분은 발효 모로미이고, 주류 제조 공정은 와인의 양조 공정이다.That is, the present invention is the genus Lactobacillus which has an immunomodulatory effect contained in the microorganism-containing component of the liquor production process. In particular, the microbial-containing component is fermented moromi, and the liquor manufacturing process is a wine brewing process.

또, 본 발명은, 면역 조절 작용을 갖는 락토바실러스·플란타룸 (Lactobacillus plantarum) 또는 락토바실러스·브레비스 (Lactobacillus brevis) 이다.Moreover, this invention is Lactobacillus plantarum or Lactobacillus brevis which has an immunomodulatory effect.

또한 본 발명은, 면역 조절 작용을 갖는 락토바실러스·플란타룸이 락토바실러스·플란타룸 SAM2446 주 (FERM ABP-10438) 또는 락토바실러스·브레비스가 락토바실러스·브레비스 SAM2447 주 (FERM ABP-10439) 이다.In the present invention, Lactobacillus plantarum having an immunomodulatory action is Lactobacillus plantarum SAM2446 strain (FERM ABP-10438) or Lactobacillus brevis Lactobacillus brevis SAM2447 strain (FERM ABP-10439) .

또, 본 발명은, 주류 제조 공정의 미생물 함유 성분에 포함되는, 면역 조절 작용을 갖는 락토바실러스속균을 함유하는 면역 조절 작용을 갖는 조성물이다. 특히 상기 조성물은 음식품 또는 의약품이다.Moreover, this invention is a composition which has an immunomodulatory effect containing the Lactobacillus genus which has an immunomodulatory effect contained in the microorganism containing component of a liquor manufacturing process. In particular, the composition is a food or beverage or a medicine.

또한 본 발명은, 주류 제조 공정의 미생물 함유 성분으로부터 미생물을 분리하는 공정, 분리된 각 미생물의 면역 조절 작용을 측정하는 공정, 및 면역 조절 작용이 강한 유산균을 선발하는 공정을 포함하는, 면역 조절 작용을 갖는 유산균을 분리하는 방법이다. 특히, 미생물 함유 성분은 발효 모로미이고, 주류 제조 공정은 와인의 양조 공정이다.In addition, the present invention includes a step of separating the microorganisms from the microorganism-containing component of the liquor manufacturing process, the step of measuring the immunomodulatory action of each separated microorganism, and the step of selecting a lactic acid bacterium with a strong immunomodulatory action, immunomodulatory action It is a method of separating the lactic acid bacteria having. In particular, the microbial-containing component is fermented moromi, and the liquor manufacturing process is a wine brewing process.

도 1 은 12 종류의 유산균을 마우스에 복강 내 투여한 후의 혈청 IL-12 농도를 측정한 결과를 나타내는 그래프이다.1 is a graph showing the results of measurement of serum IL-12 concentration after intraperitoneal administration of 12 kinds of lactic acid bacteria to mice.

도 2 는 해독된 SAM2446 주의 16S rRNA 유전자 배열을 나타낸다. 하선부는, 락토바실러스·플란타룸 JCM1149T (ATCC14917) 와의 배열의 상이 부분을 나타낸다.2 shows the 16S rRNA gene sequence of the translated SAM2446 strain. The underlined portion shows a portion different from the arrangement with Lactobacillus plantarum JCM1149T (ATCC14917).

도 3 은 해독된 SAM2447 주의 16S rRNA 유전자 배열을 나타낸다. 하선부는, 락토바실러스·브레비스 JCM1059T (ATCC14869) 와의 배열의 상이 부분을 나타낸다.3 shows the 16S rRNA gene sequence of the translated SAM2447 strain. The underlined portion represents a portion different from the arrangement with Lactobacillus brevis JCM1059T (ATCC14869).

도 4 는 NK 활성의 측정 결과를 나타낸다.4 shows measurement results of NK activity.

도 5 는 IFN-γ 농도의 측정 결과를 나타낸다.5 shows measurement results of IFN-γ concentration.

발명을 실시하기Implement the invention 위한 최선의 형태 Best form for

<락토바실러스속균><Lactobacillus genus>

락토바실러스속균은, 유산균의 일종으로서, 본 발명은 주류 제조 공정의 미생물 함유 성분에 포함되는, 면역 조절 작용을 갖는 락토바실러스속균이다. 락토바실러스속균은, 락토바실러스·플란타룸 (Lactobacillus plantarum), 또는 락토바실러스·브레비스 (Lactobacillus brevis) 에 속하는 락토바실러스속균이 바람직하다.The Lactobacillus genus is a kind of lactic acid bacteria, and the present invention is a Lactobacillus genus which has an immunomodulatory effect contained in the microorganism-containing component of the liquor production process. As for Lactobacillus genus, Lactobacillus plantarum or Lactobacillus genus belonging to Lactobacillus brevis is preferable.

락토바실러스속균의 균학적 성질은, 버지스·매뉴얼·오브·시스테마틱·박테리올로지 (BERGEY'S MANUAL of Systematic Bacteriology) (제 1 권 1984년, 제 2 권 1986년, 제 3 권 1989년, 제 4 권 1989년) 에 기재되어 있다.The bacteriological properties of the genus Lactobacillus are BERGEY'S MANUAL of Systematic Bacteriology (Vol. 1, 1984, Vol. 2, 1986, Vol. 1989, Vol. 4, 1989).

본 발명에 관련된 유산균의 대표적인 것으로서, 락토바실러스·플란타룸 SAM2446 주, 및 락토바실러스·브레비스 SAM2447 주를 들 수 있다. 본 균은 독립 행정 법인 산업 기술 종합 연구소, 특허 생물 기탁 센터 (일본 이바라키현 츠쿠바시 히가시 1쵸메 1반치 1 츄오 다이6) 에 기탁되어 있고, 그 수탁 번호는 FERM ABP-10438, 및 FERM ABP-10439 이다. 락토바실러스·플란타룸 SAM2446 주 (FERM ABP-10438) 의 균학적 성질을 표 1 에, 락토바실러스·브레비스 SAM2447 주 (FERM ABP-10439) 의 균학적 성질을 표 2 에 나타낸다.As a typical example of the lactic acid bacteria which concerns on this invention, Lactobacillus plantarum SAM2446 strain and Lactobacillus brevis SAM2447 strain are mentioned. This bacterium has been deposited in an independent administrative corporation, the Industrial Technology Research Institute, and the Patent Biological Deposit Center (1 Chuo Dai 6, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan) .The accession numbers are FERM ABP-10438 and FERM ABP-10439. to be. The microbiological properties of the Lactobacillus plantarum SAM2446 strain (FERM ABP-10438) are shown in Table 1, and the microbial properties of the Lactobacillus Brevis SAM2447 strain (FERM ABP-10439) are shown in Table 2.

[표 1]TABLE 1

락토바실러스·플란타룸 SAM2446 주 (FERM ABP-10438)Lactobacillus Planta Room SAM2446 Note (FERM ABP-10438)

세포 형태 포자 그램 염색성 운동성 아포 카탈라아제 반응 15℃ 에서의 생육 45℃ 에서의 생육 당의 자화성 (양성 : +, 음성 : -, 약양성 : w) 글리세롤 D-아라비노오스 L-아라비노오스 리보오스 D-자일로오스 L-자일로오스 갈락토오스 글루코오스 프룩토오스 만노오스 람노오스 만니톨 소르비톨 α-메틸-D-만노시드 α-메틸-D-글루코시드 셀로비오스 락토오스 멜리비오스 트레할로스 라피노오스 자일리톨Cell Form Spore Gram Staining Motile Apocatalase Reaction Growth at 15 ° C Magnetization of Growth Sugar at 45 ° C (positive: +, negative:-, weakly positive: w) Glycerol D-Arabinose L-Arabinose Ribose D- Xylose L-Xylose Galactose Glucose Fructose Mannose Rhamnose Mannitol Sorbitol α-Methyl-D-Mannoside α-Methyl-D-Glucoside Cellobiose Lactose Trehalose Raffinose Xylitol 간균 형성하지 않음 양성 없음 없음 음성 양호 발육하지 않음 - - + + - - + + + + - + + - - + + - - - -Bacillus not formed Positive None None Negative Good Not developed--+ +--+ + + +-+ +--+ +----

[표 2]TABLE 2

락토바실러스·브레비스 SAM2447 주 (FERM ABP-10439)Lactobacillus Brevis SAM2447 Note (FERM ABP-10439)

세포 형태 포자 그램 염색성 운동성 아포 카탈라아제 반응 15℃ 에서의 생육 45℃ 에서의 생육 당의 자화성 (양성 : +, 음성 : -, 약양성 : w) 글리세롤 D-아라비노오스 L-아라비노오스 리보오스 D-자일로오스 L-자일로오스 갈락토오스 글루코오스 프룩토오스 만노오스 람노오스 만니톨 소르비톨 α-메틸-D-만노시드 α-메틸-D-글루코시드 셀로비오스 락토오스 멜리비오스 트레할로스 라피노오스 자일리톨Cell Form Spore Gram Staining Motile Apocatalase Reaction Growth at 15 ° C Magnetization of Growth Sugar at 45 ° C (positive: +, negative:-, weakly positive: w) Glycerol D-Arabinose L-Arabinose Ribose D- Xylose L-Xylose Galactose Glucose Fructose Mannose Rhamnose Mannitol Sorbitol α-Methyl-D-Mannoside α-Methyl-D-Glucoside Cellobiose Lactose Trehalose Raffinose Xylitol 간균 형성하지 않음 양성 없음 없음 음성 양호 발육하지 않음 - w + + - - + + + - - w - - w - - - - - -Bacillus not formed Positive None None Negative Good Not developed-w + +--+ + +--w--w------

본 발명에 있어서,「면역 조절 작용을 갖는」이란, 정상 상태나 면역 기능이 저하되어 있는 경우에 이것을 활성화하는 작용 (면역 부활 작용), 또는 면역 기능이 과도하게 항진되어 있을 때에 이것을 억제하여 적절한 면역 상태로 하는 작용 (면역 억제 작용), 세포성 면역과 액성 면역의 밸런스를 최적화하는 작용 중 적어도 어느 일방의 작용을 갖는 것을 의미한다. 면역 조절 작용으로는, 예를 들어, 사이토카인 산생의 촉진 또는 억제 작용, 림프구 활성화 작용, NK (내츄럴·킬러) 활성 증강 작용, Th1/Th2 밸런스 개선 작용, 면역 저하 억제 작용, 항알레르기 작용 등을 들 수 있는데, 이들로 한정되는 것은 아니다.In the present invention, the term "having an immunomodulatory action" refers to an action of activating this when a steady state or immune function is lowered (immune reactivation action) or to suppressing an appropriate immune function when the immune function is excessively advanced. It means having the action of at least one of the effect of making a state (immunosuppressive action) and the effect of optimizing the balance between cellular immunity and humoral immunity. Examples of immunomodulatory actions include promoting or inhibiting cytokine production, activating lymphocytes, enhancing NK (natural killer) activity, improving Th1 / Th2 balance, suppressing immune lowering, antiallergic action, and the like. Although it may be mentioned, It is not limited to these.

본 발명의 락토바실러스속균은, 주류 제조 공정의 미생물 함유 성분으로부터 미생물을 분리하고, 분리된 각 미생물의 면역 조절 작용을 측정하고, 면역 조절 작용이 강한 균을 선발함으로써 얻을 수 있다.The Lactobacillus genus of the present invention can be obtained by separating microorganisms from the microorganism-containing component of the liquor production process, measuring the immunomodulatory action of each microorganism separated, and selecting a bacterium with a strong immunomodulatory action.

본 발명에 있어서「주류 제조 공정」이란, 주류 제조에 관련된 일련의 공정이다. 여기서,「주류」에는, 와인 등의 과실주, 맥주나 발포주 등의 맥아 발효 음료, 청주, 위스키 등의 증류주 등도 포함된다.In the present invention, the "liquor production process" is a series of processes related to alcohol production. Here, the "liquor" includes fruit wines such as wine, malt fermented beverages such as beer and sparkling wine, distilled wines such as sake, whiskey and the like.

미생물 함유 성분이란, 주류 제조 공정 중에서 미생물이 존재하는 성분을 말하고, 예를 들어 주입 원료, 발효요 등이다.The microorganism-containing component refers to a component in which microorganisms exist in the liquor production process, and is, for example, an injection raw material or fermented urine.

<조성물><Composition>

본 발명에 관련된 락토바실러스속균은, 당해 락토바실러스속균을 함유하는 조성물로서 이용할 수 있다. 특히, 면역 조절 작용을 갖는 약학적 조성물로서의 이용이 바람직하다.The Lactobacillus genus according to the present invention can be used as a composition containing the Lactobacillus genus. In particular, use as a pharmaceutical composition having an immunomodulatory action is preferable.

상기 조성물에 함유되는 락토바실러스속균으로는, 락토바실러스속균 (생균 및 사균), 락토바실러스속균 함유물, 락토바실러스속균 발효물, 락토바실러스속균 처리물 등을 들 수 있다. 생균은 당해 락토바실러스속균 배양액 등의 락토바실러스속균 함유물로부터 얻을 수 있다. 사균은, 예를 들어, 생균에 대해 가열, 자외선 조사, 포르말린 처리 등을 실시함으로써 얻을 수 있다. 얻어진 생균, 사균은, 또한 마쇄나 파쇄 등에 의해 처리물로 할 수 있다.As Lactobacillus genus contained in the said composition, Lactobacillus genus (live and dead bacteria), Lactobacillus genus containing material, Lactobacillus genus fermentation product, Lactobacillus genus processed material, etc. are mentioned. Live bacteria can be obtained from Lactobacillus bacteria-containing substances such as the Lactobacillus bacteria culture medium. The dead bacteria can be obtained by, for example, heating, ultraviolet irradiation, formalin treatment or the like to live bacteria. The obtained live bacteria and dead bacteria can be further treated by grinding or crushing.

즉, 본 발명의 락토바실러스속균을 이용하는 락토바실러스속균 함유 조성물은, 본 발명에 관련된 락토바실러스속균, 당해 락토바실러스속균 함유물, 당해 락 토바실러스속균 발효물, 당해 락토바실러스속균 처리물 중 적어도 어느 하나를 함유하고 있으면 된다. 상기 락토바실러스속균 함유물로는, 예를 들어, 락토바실러스속균 현탁액, 락토바실러스속균 배양물 (균체, 배양 상청, 배지 성분을 포함한다), 락토바실러스속균 배양액 (균 배양물로부터 고형분을 제거한 것), 락토바실러스속균 발효유 (락토바실러스속균 음료, 산유, 요구르트 등) 를 들 수 있다. 락토바실러스속에 속하는 미생물의 배양에 사용하는 배지로는, 락토바실러스속에 속하는 미생물을 생육할 수 있는 배지이면 특별히 제한없이 사용할 수 있다. 또, 락토바실러스속균 처리물로는, 예를 들어, 마쇄물, 파쇄물, 액상물 (추출액 등), 농축물, 페이스트화물, 건조물 (분무 건조물, 동결 건조물, 진공 건조물, 드럼 건조물 등), 희석물 등을 들 수 있다. 또, 본 발명에 관련된 락토바실러스·플란타룸 SAM2446 주, 및 락토바실러스·브레비스 SAM2447 주는 와인 양조 공정으로부터 분리된 것이므로, 예를 들어, 과채류나 곡류를 본 발명에 관련된 락토바실러스속균에 의해 발효시킨 발효물을 함유하는 것도 본 발명의 락토바실러스속균을 이용하는 유산균 함유 조성물의 1 형태로서 바람직하다. 또한, 상기 서술한 바와 같이, 본 발명에 관련된 락토바실러스속균은 원래 식품으로부터 분리된 것이기 때문에 안전성은 높다.That is, the Lactobacillus genus containing composition using the Lactobacillus genus of this invention is at least any one of the Lactobacillus genus which concerns on this invention, the said Lactobacillus genus containing substance, the said Lactobacillus genus fermentation product, and the said Lactobacillus genus processed material. What is necessary is just to contain. Examples of the Lactobacillus genus-containing material include, for example, Lactobacillus genus suspensions, Lactobacillus genus cultures (including microorganisms, culture supernatants, and media components), and Lactobacillus genus culture medium (solids removed from the bacterial cultures). And Lactobacillus genus fermented milk (Lactobacillus genus beverage, acid oil, yoghurt, etc.) are mentioned. As a medium used for culturing microorganisms belonging to the genus Lactobacillus, any medium that can grow microorganisms belonging to the genus Lactobacillus can be used without particular limitation. As the Lactobacillus fungus treated material, for example, ground products, crushed products, liquid products (extracts, etc.), concentrates, pastes, dried products (spray dried products, freeze-dried products, vacuum dried products, drum dried products, etc.), dilutions Etc. can be mentioned. In addition, since the Lactobacillus plantarum SAM2446 strain and the Lactobacillus brevis SAM2447 strain which are related to this invention are isolate | separated from the wine brewing process, for example, the fermentation which fermented vegetables and cereals by the Lactobacillus genus bacterium which concerns on this invention was carried out. It is also preferable to contain water as one form of the lactic acid bacteria containing composition using the Lactobacillus genus of this invention. Moreover, as mentioned above, since the Lactobacillus genus concerning this invention was isolate | separated from the original foodstuff, safety is high.

본 발명의 락토바실러스속균을 이용하는 락토바실러스속균 함유 조성물은, 면역 조절 작용을 갖는 약학적 조성물로서 사용하는 것이 바람직하다. 예를 들어, 음식품, 의약품, 사료 등을 들 수 있다. 음식품에 사용하는 경우에는, 면역 조절 작용을 갖는 건강 식품으로서 실시하는 것이 바람직하다. 또, 공지된 감미료, 산미료, 비타민 등의 각종 성분과 혼합하여 사용자의 기호에 맞는 제품으로 하면 된다. 예를 들어, 정제, 캡슐제, 드링크제, 트로키, 껌, 요구르트 등의 유제품, 아이스크림, 음료, 알코올 음료, 조미료, 가공 식품, 디저트류, 과자 등의 형태로 제공할 수 있다.It is preferable to use the Lactobacillus genus containing composition using the Lactobacillus genus of this invention as a pharmaceutical composition which has an immunomodulatory effect. For example, food and drink, medicine, feed, etc. are mentioned. When used for food and drink, it is preferable to carry out as a health food which has an immunomodulatory effect. Moreover, what is necessary is just to mix with various components, such as a well-known sweetener, an acidulant, and a vitamin, and to make it the product suited to a user's preference. For example, it can be provided in the form of dairy products such as tablets, capsules, drinks, troches, gums, yogurts, ice creams, beverages, alcoholic beverages, seasonings, processed foods, desserts, confectionery.

또, 의약품에 사용하는 경우에는, 주약에 부형제, 결합제, 붕괴제, 활택제, 교미 교취제, 용해 보조제, 현탁제, 코팅제 등의 의약의 제제 기술 분야에 있어서 통상 사용하는 공지된 보조제를 사용하여 제제화할 수 있다. 제형으로는, 정제, 캡슐제, 과립제, 산제, 시럽제, 좌제, 주사제 등을 들 수 있고, 특별히 한정되는 것은 아니다. 본 의약품의 투여 경로로는, 예를 들어, 경구 투여, 직장 투여, 경장 투여 등을 들 수 있는데, 특별히 한정되는 것은 아니다.Moreover, when used for pharmaceuticals, it is formulated using the well-known adjuvant normally used in the art of manufacture of medicines, such as excipient, binder, disintegrant, glidant, copper, dissolution aid, suspending agent, coating agent, etc. in a main medicine. can do. Examples of the formulation include tablets, capsules, granules, powders, syrups, suppositories, injections, and the like, and are not particularly limited. Examples of the route of administration of the drug include oral administration, rectal administration, and enteral administration, but are not particularly limited.

사료에 사용되는 경우에는, 예를 들어, 펫 푸드, 혹은 가축용 사료 등의 원료에 균체를 혼합할 수 있다. 혹은, 시판되고 있는 동물용 사료에 첨가해도 된다. 그 중에서도, 개나 고양이, 혹은 닭의 조류 등의 식이에 바람직하게 사용할 수 있다.When used for feed, the cells can be mixed with raw materials such as pet food or livestock feed. Or you may add to the commercial animal feed. Especially, it can use suitably for diet, such as a dog, a cat, or a bird of a chicken.

<면역 조절 작용을 갖는 유산균의 분리 방법><Isolation method of lactic acid bacteria having immunomodulatory action>

면역 조절 작용의 측정은, 당업자에게 알려지는 방법 중 어느 것이어도 된다. 일례를 들면, Th1 형 사이토카인의 인터류킨 12 (이하「IL-12」라고 약기한다) 유도능 시험으로, 마우스로부터 조제한 복강 매크로파지에 대한 인 비트로에서의 처리에 의한 IL-12 의 산생량을 측정함으로써 실시한다.The measurement of an immunomodulatory effect may be any of methods known to those skilled in the art. For example, in the interleukin 12 (hereinafter abbreviated as "IL-12") induction test of the Th1 cytokine, by measuring the amount of IL-12 produced by in vitro treatment of peritoneal macrophages prepared from mice, Conduct.

면역 조절 작용이 강한 유산균의 선발은, 상기 면역 조절 작용의 측정에 의 해, 면역 조절 작용이 높은 균주를 선발함으로써 실시할 수 있다. 선발의 기준으로는, 공지된 면역 조절 작용이 알려져 있는 유산균과의 비교, 컨트롤과의 비교를 실시하거나, 혹은 평균값과 비교하는 등 적절히 설정하면 된다. 또, 면역 조절 작용 외에, 일반적인 유산균용 배지 중에서 양호한 증식을 나타내는지의 여부를 선발 기준에 추가할 수 있다.Selection of lactic acid bacteria having a strong immunomodulatory effect can be performed by selecting strains having a high immunomodulatory effect by measuring the immunomodulatory effect. What is necessary is just to set suitably as a selection criteria, for example, the comparison with a lactic acid bacterium for which a known immunomodulatory effect is known, a comparison with a control, or a mean value. In addition to the immunomodulatory action, it is possible to add to the selection criteria whether or not good proliferation is exhibited in the medium for lactic acid bacteria.

본 발명에 있어서「주류 제조 공정」이란, 주류 제조에 관련된 일련의 공정이다. 여기서,「주류」에는, 와인 등의 과실주, 맥주나 발포주 등의 맥아 발효 음료, 청주 위스키 등의 증류주 등도 포함된다.In the present invention, the "liquor production process" is a series of processes related to alcohol production. Here, the "liquor" includes fruit wines such as wine, malt fermentation drinks such as beer and sparkling wine, distilled wines such as sake whiskey, and the like.

미생물 함유 성분이란, 주류 제조 공정 중에서, 미생물이 존재하는 성분을 말하고, 예를 들어 주입 원료, 발효요 등이다.The microorganism-containing component refers to a component in which microorganisms exist in the liquor production process, and is, for example, an injection raw material or fermented urine.

분리원으로서, 상기 와인의 양조 공정의 발효 모로미 등을 사용함으로써, 매우 효율적으로 면역 조절 작용을 갖는 유산균을 분리할 수 있다.As a separation source, by using a fermented moromi or the like in the wine brewing step, lactic acid bacteria having an immunomodulatory effect can be separated very efficiently.

본 발명을 실시예에 의해 더욱 상세하게 설명하는데, 본 발명은 상기 서술한 각 실시형태에 한정되는 것은 아니며, 청구항에 나타낸 범위에서 다양한 변경이 가능하고, 상이한 실시형태에 각각 개시된 기술적 수단을 적절히 조합하여 얻어지는 실시형태에 대해서도 본 발명의 기술적 범위에 포함된다.EXAMPLES The present invention will be described in more detail with reference to Examples, but the present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and combinations of the technical means disclosed in the different embodiments are appropriately combined. Embodiment obtained by making is also included in the technical scope of this invention.

실시예 1Example 1

[사용 유산균주][Use Lactobacillus]

와인 양조료로부터, 유산균을 104 주 분리하고, Th1 형 면역 부활능이 높은 균주를 선택하기 위해 인터류킨 12 (IL-12) 유도능을 비교하는 실험에 제공하였다.From wine brewing, lactic acid bacteria were isolated for 104 weeks and used in experiments comparing interleukin 12 (IL-12) inducing ability to select strains having high Th1 type immune activating ability.

[인 비트로 자극에 의한 IL-12 유도][Induction of IL-12 by In Vitro Stimulation]

BALB/c 마우스 (생후 7 주·수컷) 에 4.05% 티오글리콜레이트를 복강 내 투여하고, 4 일 후에 PBS 를 사용하여 복강 내 매크로파지를 회수하고, 10% FBS 를 포함하는 RPMI 배지를 사용하여 2 × 106cells/㎖ 로 조정한 후, 24 웰 (well) 플레이트에 0.5㎖ 씩 뿌렸다. 각 웰에 각 균주의 가열 사균 (1㎍/㎖) 을 첨가하여 24 시간 배양 후의 배양 상청 중의 IL-12 농도를 측정하였다. 컨트롤은, 사균 무첨가인 것으로 하였다. IL-12 의 활성형은 p35 와 p40 의 서브유닛이 결합된 p70 이기 때문에, IL-12 (p70) 를 측정하였다. 또한 IL-12 의 측정에는 OptEIA mouse IL-12 측정 키트 (BD Pharmingen 사 제조) 를 사용하였다.Intraperitoneal administration of 4.05% thioglycolate to BALB / c mice (7 weeks old male), 4 days later, intraperitoneal macrophages were recovered using PBS, and 2 × using RPMI medium containing 10% FBS. After adjusting to 10 6 cells / ml, it was sprinkled 0.5 ml in a 24 well plate. Heat killing bacteria (1 µg / ml) of each strain was added to each well, and IL-12 concentration in the culture supernatant after 24 hours of culture was measured. The control was supposed to be dead bacteria free. IL-12 (p70) was measured because the active form of IL-12 is p70 to which p35 and p40 subunits are bound. In addition, the OptEIA mouse IL-12 measurement kit (made by BD Pharmingen) was used for the measurement of IL-12.

분리한 유산균 104 주에 대해 인 비트로에 의한 IL-12 산생능을 평가한 결과, IL-12 산생능을 갖는 균주가 많이 포함되어 있었다. 이들 몇 가지에 대해, IL-12 산생능은, 면역 조절 작용이 알려져 있는 유산균과 비교해도 충분히 높은 활성을 갖고 있는 것으로 생각되었다. 이 중, 컨트롤에 대해 10 배를 초과하는 IL-12 산생능을 갖는 12 주에 대해, 높은 IL-12 산생능을 갖고 있는 주로서 선택하였다.As a result of evaluating IL-12 acid production by in vitro for the 104 strains of isolated lactic acid bacteria, many strains having IL-12 acid production were included. For some of these, IL-12 production was thought to have a sufficiently high activity compared with lactic acid bacteria whose immunomodulatory activity is known. Of these, for 12 weeks having a IL-12 production capacity more than 10 times the control, it was selected as a week having a high IL-12 production capacity.

컨트롤에 대해 10 배라는 매우 높은 기준을 고려하면, 이 확률은, 상당히 높은 값인 것으로 생각되고, 와인 양조료는, 면역 조절 유산균의 분리원으로서 매우 유용한 것으로 생각되었다.Considering a very high criterion of 10 times for the control, this probability was considered to be a fairly high value, and wine brewing was considered to be very useful as a source of immunoregulatory lactic acid bacteria.

[표 3]TABLE 3

Figure 112008036638288-PCT00001
Figure 112008036638288-PCT00001

실시예 2Example 2

[사용 유산균주][Use Lactobacillus]

실시예 1 에서 선택한 유산균 12 주를, 인 비보 자극에 의한 IL-12 유도능을 비교하는 실험에 제공하였다.Twelve weeks of lactic acid bacteria selected in Example 1 were used for the experiment comparing IL-12 inducing ability by in vivo stimulation.

[인 비보 자극에 의한 IL-12 유도][Induction of IL-12 by In vivo Stimulation]

BALB/c 마우스 (생후 7 주·수컷) 에 상기 각 균주의 가열 사균 (500㎍/0.2㎖/mouse) 의 현탁액 (용매 : 생리적 식염수) 을 복강 내 투여하고, 6 시간 후에 경추 탈구 후, 심장으로부터 채혈하였다. 또한, 컨트롤의 마우스에는 생리적 식염수를 등량 투여하였다. 채혈 후, 원심 분리에 의해 혈청을 채취하고, 혈청 중의 IL-12 농도를 OptEIA mouse IL-12 측정 키트 (BD Pharmingen 사 제조) 를 사용하여 측정하였다.BALB / c mice (7 weeks, males) were administered intraperitoneally with a suspension (solvent: physiological saline) of heated bacteria (500 µg / 0.2 ml / mouse) of each strain, and after 6 hours, after cervical dislocation, Blood was collected. In addition, the control mice were dosed with an equal amount of physiological saline. After blood collection, serum was collected by centrifugation, and IL-12 concentration in serum was measured using an OptEIA mouse IL-12 measurement kit (manufactured by BD Pharmingen).

결과를 도 1 에 나타냈다. 혈청 IL-12 농도는, A 주와 L 주에서 특히 강하였다. 그래서, 이들을 각각 SAM2446 주, SAM2447 주라고 명명하였다.The results are shown in FIG. Serum IL-12 concentration was especially strong in A and L weeks. Thus, they were named as SAM2446 and SAM2447, respectively.

실시예 3 SAM2446 주, SAM2447 주의 동정 (同定)Example 3 Identification of SAM2446 strain and SAM2447 strain

SAM2446 주의 균학적 성질의 검토 결과를 표 1 에, SAM2447 주의 균학적 성질의 검토 결과를 표 2 에 나타냈다.The results of the examination of the bacteriological properties of SAM2446 strains are shown in Table 1, and the results of the examination of the bacteriological properties of SAM2447 strains are shown in Table 2.

이들의 검토 결과에 의해, SAM2446 주는 락토바실러스·플란타룸으로, SAM2447 주는 락토바실러스·브레비스로 추정되었다. 락토바실러스·플란타룸은 소르비톨의 자화성이 있는 것과 없는 것의 양자가 알려져 있는데, 표 1 에 나타낸 바와 같이, SAM2446 주는 소르비톨의 자화성을 갖는 유산균종이다. 또, 락토바실러스·브레비스는 D-자일로오스의 자화성이 있는 것과 없는 것의 양자가 알려져 있는데, 표 2 에 나타낸 바와 같이, SAM2447 주는 D-자일로오스의 자화성을 가지지 않는 유산균종이다.Based on the results of these studies, the SAM2446 strain was estimated to be Lactobacillus plantarum and the SAM2447 strain was lactobacillus brevis. Lactobacillus plantarum is known both with and without magnetization of sorbitol. As shown in Table 1, SAM2446 strain is a lactic acid bacterium having magnetization of sorbitol. Incidentally, both Lactobacillus and Brevis are known to have D-xylose magnetization and non-magnetic properties. As shown in Table 2, the SAM2447 strain is a lactic acid species that does not have D-xylose magnetization.

SAM2446 주 및 SAM2447 주로부터 DNA 를 추출 후, 16S rRNA 유전자 영역에 대해 해독을 실시하였다. 해독된 SAM2446 주의 16S rRNA 유전자 배열 (배열 번호 1) 은, 락토바실러스·플란타룸 JCM1149T (ATCC14917) 의 16S rRNA 유전자 배열과 99% 상동성이 얻어졌기 때문에, 락토바실러스·플란타룸으로 동정하였다 (도 2). 또, SAM2447 주의 16S rRNA 유전자 배열 (배열 번호 2) 은, 락토바실러스·브레비스 JCM1059T (ATCC14869) 의 16S rRNA 유전자 배열과 97% 상동성이 얻어 졌기 때문에, 락토바실러스·브레비스로 동정하였다 (도 3).DNA was extracted from the SAM2446 and SAM2447 strains, followed by translation of the 16S rRNA gene region. The 16S rRNA gene sequence (SEQ ID NO: 1) of the decoded SAM2446 strain was identified as Lactobacillus plantarum because 99% homology with the 16S rRNA gene sequence of Lactobacillus plantarum JCM1149T (ATCC14917) was obtained ( 2). The 16S rRNA gene sequence of the SAM2447 strain (SEQ ID NO: 2) was identified as Lactobacillus brevis because 97% homology with the 16S rRNA gene sequence of Lactobacillus Brevis JCM1059T (ATCC14869) was obtained (Fig. 3).

실시예 4 경구 섭취에 의한 면역 부활 및 면역 조절 작용Example 4 Immune Revitalization and Immune Regulation by Oral Ingestion

C57BL/6 마우스 (니혼 찰스리버 주식회사, 생후 7 주·수컷, 1 군 4 마리) 에, SAM2446 주, SAM2447 주 (사균체) 를 1 주간 음수 (0.2㎎/day 상당) 로 자유롭게 섭취시켰다. 또, SAM2446 주, SAM2447 주를 섭취시키지 않는 대조군을 형성하였다. 1 주일 후에 비장을 적출하였다. 적출한 비장으로부터 통상적인 방법에 따라 비장 세포를 조제하고, NK 활성과, 사이토카인 산생능을 측정하였다. NK 활성은 비장 세포와 타겟 세포인 Yac-1 을 200 : 1, 또는 100 : 1 의 비율로 4 시간 공배양하고, 비장 세포의 세포 장해성 활성을 플로우 사이토메트리를 사용한 PINK 법으로 측정하였다. 또, 비장 세포 (5 × 106 개/㎖) 에 콘카나발린 A (나카라이테스크) 2.5㎎/㎖ 를 24 시간 작용시키고, 배양 상청 중에 산생되는 IFN-γ 농도를 ELISA 법 (OptEIA (BD BioScience)) 으로 측정하였다.C57BL / 6 mice (Nihon Charles River Co., Ltd., 7 weeks male and 4 males, 1 group) were freely ingested with SAM2446 strain and SAM2447 strain (killing bodies) for 1 week in negative water (equivalent to 0.2 mg / day). Moreover, the control group which does not ingest SAM2446 week and SAM2447 week was formed. After one week the spleens were removed. Splenocytes were prepared from the spleens removed according to a conventional method, and NK activity and cytokine acid production were measured. NK activity was co-cultured with spleen cells and target cells Yac-1 at a ratio of 200: 1 or 100: 1 for 4 hours, and cytotoxic activity of splenocytes was measured by PINK method using flow cytometry. In addition, 2.5 mg / ml of concanavalin A (Nakaraitesk) was allowed to act on spleen cells (5 × 10 6 cells / ml) for 24 hours, and IFN-γ concentration produced in the culture supernatant was determined by ELISA method (OptEIA (BD BioScience). Was measured).

NK 활성의 측정 결과를 도 4 에 나타냈다. 도 4 중, NK 활성 (%) 은 마우스 비장 세포의 Yac-1 에 대한 세포 상해성을 나타내고, E : T 비는 반응시킨 비장 세포수 : Yac-1 수를 나타낸다. 도 4 로부터 분명한 바와 같이, SAM2446 주, SAM2447 주를 경구 섭취한 마우스로부터 조제한 비장 세포의 NK 활성은 대조군 (control) 과 비교하여 현저하게 상승되어 있었다.The measurement result of NK activity is shown in FIG. In FIG. 4, NK activity (%) shows cytotoxicity with respect to Yac-1 of mouse spleen cells, and E: T ratio shows the spleen cell number: Yac-1 number which reacted. As is apparent from FIG. 4, the NK activity of spleen cells prepared from mice orally ingested with SAM2446 and SAM2447 strains was significantly increased as compared with the control.

IFN-γ 농도의 측정 결과를 도 5 에 나타냈다. 도 5 로부터 분명한 바와 같이, SAM2446 주, SAM2447 주를 경구 섭취한 마우스로부터 조제한 비장 세포에서 는 대조군 (control) 과 비교하여 IFN-γ 농도가 분명하게 증가되어 있었다.The measurement result of IFN- (gamma) concentration is shown in FIG. As is apparent from FIG. 5, the spleen cells prepared from mice orally ingested with SAM2446 and SAM2447 were significantly increased in IFN-γ concentrations as compared with the control.

이상과 같이, SAM2446 주, SAM2447 주를 경구 섭취한 마우스에 있어서, NK 활성이 증가하고, Th1 형 사이토카인인 IFN-γ 의 산생이 촉진되었다. 따라서, SAM2446 주, SAM2447 주는, 면역 부활 작용 및 면역 조절 작용을 갖는 것이 분명해졌다.As mentioned above, NK activity increased and the production | generation of IFN- (gamma) which is Th1-type cytokine was accelerated | stimulated in the mouse which orally ingested SAM2446 week and SAM2447 week. Therefore, it became clear that the SAM2446 strain and the SAM2447 strain have an immunostimulating action and an immunomodulatory action.

실시예 5 두유 발효물의 제조Example 5 Preparation of Soymilk Fermentation

SAM2446 주 발효 균액을, 다른 미생물 오염을 받지 않도록 시판되고 있는 두유에 첨가하고, 30℃ 에서 24 시간 정치 (靜置) 하여 두유 발효물을 얻었다. 발효는 신속하게 진행되어, 산도 상승 및 pH 의 저하를 볼 수 있었다. 얻어진 발효물은, 종래의 식물 발효품에 비교하여 불쾌한 발효취 등을 나타내지 않고, 관능적으로 우수한 향미를 나타냈다.The SAM2446 main fermentation bacterial solution was added to commercially available soymilk so as not to be subjected to other microbial contamination, and left standing at 30 ° C for 24 hours to obtain a soymilk fermentation product. The fermentation proceeded rapidly, and the acidity rise and pH fall were seen. The obtained fermented product did not exhibit an unpleasant fermentation odor or the like compared with the conventional plant fermented product, and exhibited a sensory excellent flavor.

실시예 6 유산균 분말 첨가 두유의 제조Example 6 Preparation of Soymilk Added with Lactic Acid Bacteria Powder

SAM2447 주 발효 균액을 가열 살균 후, 원심 분리 (3000rpm, 10 분) 에 의해 균체를 회수하고, 얻어진 균체에 대해 동결 건조를 실시하였다. 이 유산균 균체 분말을 시판되고 있는 두유에 첨가·교반하여 두유 중에 현탁 용해시켜 얻어지는 유산균 분말 첨가 두유는, 종래의 유산균 함유 발효 식품에 비교하여 불쾌한 발효취 등을 나타내지 않고, 관능적으로도 우수한 향미를 나타냈다.After heat sterilization of the SAM2447 main fermentation bacteria solution, the cells were collected by centrifugation (3000 rpm, 10 minutes), and the obtained cells were freeze-dried. The lactic acid bacteria powder-added soy milk obtained by adding and stirring the lactic acid bacteria cell powder to commercially available soymilk and suspended in soymilk exhibited a taste that was superior to conventional lactic acid bacteria-containing fermented foods without any unpleasant fermentation odor. .

실시예 7 유산균 함유 조성물의 제조Example 7 Preparation of Lactic Acid Bacteria-Containing Composition

제조예 1 : 유산균을 함유하는 의약품Preparation Example 1 Drug Containing Lactic Acid Bacteria

정제 :refine :

이하에 나타내는 방법에 의해, 유산균을 함유하는 의약품 (정제) 을 제조하였다.By the method shown below, the pharmaceutical product (tablet) containing a lactic acid bacterium was manufactured.

SAM2446 주균체 분말 50g 을, 젖당 250g 및 스테아르산마그네슘 2g 과 함께 혼합하고, 단발식 타정기로 타정함으로써, 직경 10㎜, 중량 300㎎ 의 정제를 제조하였다.50 g of SAM2446 main cell powder was mixed with 250 g of lactose and 2 g of magnesium stearate, and compressed into a single tableting machine to prepare a tablet having a diameter of 10 mm and a weight of 300 mg.

과립제 :Granules:

SAM2447 주균체 분말 100g 에, 스테아르산마그네슘 0.5g 을 첨가하고, 압축, 분쇄, 정립하고, 체질하여 20 ∼ 50 메시의 과립제를 얻었다.0.5 g of magnesium stearate was added to 100 g of SAM2447 main cell powder, which was compacted, pulverized, sieved, and sieved to obtain a granule of 20 to 50 mesh.

제조예 2 : 유산균을 함유하는 각종 음식물Preparation Example 2 Various Foods Containing Lactic Acid Bacteria

이하에 나타내는 조성으로, 유산균이 들어간 각종 음식품을 제조하였다.Various food-drinks containing the lactic acid bacterium were manufactured with the composition shown below.

[표 4]TABLE 4

엿 :Fucked:

(조성) (중량부)(Composition) (parts by weight)

분말 소르비톨 99.7Powder Sorbitol 99.7

향료 0.2Spices 0.2

SAM2446 주균체 분말 0.05SAM2446 Main Cell Powder 0.05

소르비톨시드Sorbitol seed 0.05            0.05

전체량 100.00Total amount 100.00

[표 5]TABLE 5

젤리 (커피 젤리) :Jelly (coffee jelly):

(조성) (중량부)(Composition) (parts by weight)

그래뉴당 15.015.0 per Granyu

젤라틴 1.0Gelatin 1.0

커피 엑기스 5.0Coffee Extract 5.0

물 78.93Water 78.93

SAM2447SAM2447 주균체Main cell 분말 0.07 Powder 0.07

전체량 100.00Total amount 100.00

[표 6]TABLE 6

주스 :juice :

(조성) (중량부)(Composition) (parts by weight)

냉동 농축 온주 밀감 과즙 5.0Frozen Concentrated Wenzhou Citrus Juicer 5.0

과당 포도당 액당 11.0Fructose glucose liquid 11.0

시트르산 0.2Citric Acid 0.2

L-아스코르브산 0.02L-ascorbic acid 0.02

SAM2446 주균체 분말 0.05SAM2446 Main Cell Powder 0.05

향료 0.2Spices 0.2

색소 0.1Pigment 0.1

물 83.43Water 83.43

전체량 100.00Total amount 100.00

[표 7]TABLE 7

탄산 음료 :carbonated drink :

(조성) (중량부)(Composition) (parts by weight)

그래뉴당 8.08.0 per granue

농축 레몬 과즙 1.0Concentrated Lemon Juicer 1.0

L-아스코르브산 0.10L-ascorbic acid 0.10

시트르산 0.09Citric Acid 0.09

시트르산나트륨 0.05Sodium Citrate 0.05

착색료 0.05Colorant 0.05

향료 0.15Spices 0.15

탄산수 90.55Sparkling water 90.55

SAN12447SAN12447 주균체Main cell 분말 0.01 Powder 0.01

전체량 100.00Total amount 100.00

[표 8]TABLE 8

유산균 음료 :Lactic acid bacteria drink:

(조성) (중량부)(Composition) (parts by weight)

유고형분 21% 발효유 14.76Milk Solid 21% Fermented Milk 14.76

과당 포도당 액당 13.31Fructose glucose liquid sugar 13.31

펙틴 0.5Pectin 0.5

시트르산 0.08Citric Acid 0.08

향료 0.15Spices 0.15

물 71.14Water 71.14

SAM2446SAM2446 주균체Main cell 분말 0.06 Powder 0.06

전체량 100.00Total amount 100.00

[표 9]TABLE 9

유산균 음료 :Lactic acid bacteria drink:

(조성) (중량부)(Composition) (parts by weight)

유고형분 21% 의 SAM2447 주 발효유 14.82SAM2447 main fermented milk with 21% milk solids 14.82

과당 포도당 액당 13.31Fructose glucose liquid sugar 13.31

펙틴 0.5Pectin 0.5

시트르산 0.08Citric Acid 0.08

향료 0.15Spices 0.15

물 71.14Water 71.14

전체량 100.00Total amount 100.00

[표 10]TABLE 10

커피 음료 :Coffee Drinks:

(조성) (중량부)(Composition) (parts by weight)

그래뉴당 8.08.0 per granue

탈지분유 5.0Skim milk powder 5.0

캐러멜 0.2Caramel 0.2

커피 추출물 2.0Coffee Extract 2.0

향료 0.1Spices 0.1

폴리글리세린지방산에스테르 0.05Polyglycerin Fatty Acid Ester 0.05

식염 0.05Saline 0.05

물 84.56Water 84.56

SAM2446SAM2446 주균체Main cell 분말 0.04 Powder 0.04

전체량 100.00Total amount 100.00

[표 11]TABLE 11

알코올 음료 :Alcoholic drinks:

(조성) (중량부)(Composition) (parts by weight)

50 용량% 에탄올 32.050% by volume ethanol 32.0

설탕 8.4Sugar 8.4

과즙 2.4Juicer 2.4

SAM2447 주균체 분말 0.2SAM2447 Main Cell Powder 0.2

정제수Purified water 57.0                            57.0

전체량 100.0Total amount 100.0

[표 12]TABLE 12

발포주 :Sparkling wine:

(조성) (중량부)(Composition) (parts by weight)

발포주 99.98Sparkling wine 99.98

SAM2447SAM2447 주균체Main cell 분말 0.02 Powder 0.02

전체량 100.0Total amount 100.0

제조예 3 : 유산균을 함유하는 사료Preparation Example 3 Feed containing Lactobacillus

이하에 나타내는 조성으로, 유산균이 들어간 사료를 제조하였다.The feed containing lactic acid bacteria was manufactured with the composition shown below.

[표 13]TABLE 13

사료 :feed :

(조성) (중량부)(Composition) (parts by weight)

시판 도크 푸드 99.98Commercial Dock Food 99.98

SAM2446SAM2446 주균체Main cell 분말 0.02 Powder 0.02

전체량 100.0 Total amount 100.0

본 발명의 락토바실러스속균을 섭취 등을 하면, 면역 기능을 부활화함으로써 면역 기능 저하를 억제할 수 있다. 또, 면역 기능의 밸런스를 조절함으로써 생체에 악영향을 미치는 면역 기능의 과도한 항진이나 그에 파생되는 생체에 있어서의 여러가지 불쾌 증상 등도 억제할 수 있다. 또, 본 발명에 의하면, 효율적으로 면역 조절 작용을 갖는 유산균을 얻을 수 있다.When the Lactobacillus genus bacterium of the present invention is ingested, the immune function can be suppressed by activating the immune function. In addition, by adjusting the balance of the immune function, excessive hyperactivity of the immune function adversely affecting the living body and various unpleasant symptoms in the living body derived therefrom can be suppressed. Moreover, according to this invention, the lactic acid bacterium which has an immune regulation effect can be obtained efficiently.

SEQUENCE LISTING <110> Suntory Limited <120> A novel Lactobacillus bacterium having immunomodulationg effects and a composition for immunomodulation containing it <130> YCT-1198 <160> 4 <170> PatentIn version 3.1 <210> 1 <211> 242 <212> DNA <213> Lactobacillus plantarum <400> 1 atacatgcaa gtcgaacgaa ctctggtatt gattggtgct tgcatcatga tttacatttg 60 agtgagtggc gaactggtga gtaacacgtg ggaaacctgc ccagaagcgg gggataacac 120 ctggaaacag atgctaatac cgcataacaa cttggaccgc atggtccgag tttgaaagat 180 ggcttcggct atcacttttg gatggtcccg cggcgtatta gctagatggt ggggtaacgg 240 ct 242 <210> 2 <211> 353 <212> DNA <213> Lactobacillus plantarum <400> 2 aggcggctgg ttcctaaaag gttaccccac cgactttggg tgttacaaac tctcatggtg 60 tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg cggcatgctg atccgcgatt 120 actagcgatt ccgacttcat gtaggcgagt tgcagcctac aatccgaact gagaatggct 180 ttaagagatt agcttactct cgcgagttcg caactcgttg taccatccat tgtagcacgt 240 gtgtagccca ggtcataagg ggcatgatga tttgacgtca tccccacctt cctccggttt 300 gtcaccggca gtctcaccac aatgcccaac ttaatgctgg caacagataa taa 353 <210> 3 <211> 346 <212> DNA <213> Lactobacillus brevis <400> 3 aacgctggcg gcatgcctaa tacatgcaag tcgaacgagc ttccgttgaa tgacgtgctt 60 gcactgattt taacaatgaa gcgagtggcg aactggtgag taacacgtgg gaaatctgcc 120 cagaagcagg ggataacact tggaaacagg tgctaatacc gtataacaac aaaatccgca 180 tggattttgt ttgaaaggtg gcttcggcta tcacttctgg atgatcccgc ggcgtattag 240 ttagttggtg aggtaaaggc ccaccaagac catgatgcgt agccgacctg agagggtaat 300 cgcccacatt gggactggca cacggcccat actcctacgg gaggca 346 <210> 4 <211> 524 <212> DNA <213> Lactobacillus brevis <400> 4 ctaatcatct gtcccacctt agacggctga ctcccgaagg ttatctcacc ggctttgggt 60 gttacaaact ctcatggtgt gacgggcggt gtgtacaagg cccgggaacg tattcaccgc 120 ggcatgctga tccgcgatta ctagcgattc caacttcatg taggcgagtt gcagcctaca 180 atccgaactg agaacggctt taagagatta gcttagcctc acgacttcgc gactcgttgt 240 accgtccatt gtagcacgtg tgtagcccag gtcataaggg gcatgatgat ttgacgtcat 300 ccccaccttc ctccggtttg tcaccggcag tctcaccaga gtgcccaact gaatgctggc 360 aactgataat aagggttgcg ctcgttgcgg gacttaaccc aacatctcac gacacgagct 420 gacgacaacc atgcaccacc tgtcattctg tccccgaagg gaacgtctta tctctaagat 480 tggcggaaga tgtcaagacc tggtaaggtt cattcgcgta gctt 524                          SEQUENCE LISTING <110> Suntory Limited <120> A novel Lactobacillus bacterium having immunomodulationg effects and a composition for immunomodulation containing it <130> YCT-1198 <160> 4 <170> PatentIn version 3.1 <210> 1 <211> 242 <212> DNA <213> Lactobacillus plantarum <400> 1 atacatgcaa gtcgaacgaa ctctggtatt gattggtgct tgcatcatga tttacatttg 60 agtgagtggc gaactggtga gtaacacgtg ggaaacctgc ccagaagcgg gggataacac 120 ctggaaacag atgctaatac cgcataacaa cttggaccgc atggtccgag tttgaaagat 180 ggcttcggct atcacttttg gatggtcccg cggcgtatta gctagatggt ggggtaacgg 240 ct 242 <210> 2 <211> 353 <212> DNA <213> Lactobacillus plantarum <400> 2 aggcggctgg ttcctaaaag gttaccccac cgactttggg tgttacaaac tctcatggtg 60 tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg cggcatgctg atccgcgatt 120 actagcgatt ccgacttcat gtaggcgagt tgcagcctac aatccgaact gagaatggct 180 ttaagagatt agcttactct cgcgagttcg caactcgttg taccatccat tgtagcacgt 240 gtgtagccca ggtcataagg ggcatgatga tttgacgtca tccccacctt cctccggttt 300 gtcaccggca gtctcaccac aatgcccaac ttaatgctgg caacagataa taa 353 <210> 3 <211> 346 <212> DNA <213> Lactobacillus brevis <400> 3 aacgctggcg gcatgcctaa tacatgcaag tcgaacgagc ttccgttgaa tgacgtgctt 60 gcactgattt taacaatgaa gcgagtggcg aactggtgag taacacgtgg gaaatctgcc 120 cagaagcagg ggataacact tggaaacagg tgctaatacc gtataacaac aaaatccgca 180 tggattttgt ttgaaaggtg gcttcggcta tcacttctgg atgatcccgc ggcgtattag 240 ttagttggtg aggtaaaggc ccaccaagac catgatgcgt agccgacctg agagggtaat 300 cgcccacatt gggactggca cacggcccat actcctacgg gaggca 346 <210> 4 <211> 524 <212> DNA <213> Lactobacillus brevis <400> 4 ctaatcatct gtcccacctt agacggctga ctcccgaagg ttatctcacc ggctttgggt 60 gttacaaact ctcatggtgt gacgggcggt gtgtacaagg cccgggaacg tattcaccgc 120 ggcatgctga tccgcgatta ctagcgattc caacttcatg taggcgagtt gcagcctaca 180 atccgaactg agaacggctt taagagatta gcttagcctc acgacttcgc gactcgttgt 240 accgtccatt gtagcacgtg tgtagcccag gtcataaggg gcatgatgat ttgacgtcat 300 ccccaccttc ctccggtttg tcaccggcag tctcaccaga gtgcccaact gaatgctggc 360 aactgataat aagggttgcg ctcgttgcgg gacttaaccc aacatctcac gacacgagct 420 gacgacaacc atgcaccacc tgtcattctg tccccgaagg gaacgtctta tctctaagat 480 tggcggaaga tgtcaagacc tggtaaggtt cattcgcgta gctt 524  

Claims (15)

와인 양조 공정의 발효 모로미에 함유되는, 면역 조절 작용을 갖는 락토바실러스속균.The Lactobacillus genus which has an immunomodulatory effect contained in the fermented moromi of a wine brewing process. 제 1 항에 있어서,The method of claim 1, 락토바실러스속균이 락토바실러스·플란타룸 (Lactobacillus plantarum) 인 균.The bacterium Lactobacillus is Lactobacillus plantarum. 제 1 항에 있어서,The method of claim 1, 락토바실러스속균이 락토바실러스·브레비스 (Lactobacillus brevis) 인 균.The bacterium Lactobacillus brevis is a genus of Lactobacillus. 제 2 항에 있어서,The method of claim 2, 락토바실러스·플란타룸이 소르비톨 자화성을 갖는 것인 균.A bacterium in which Lactobacillus plantarum has sorbitol magnetization. 제 3 항에 있어서,The method of claim 3, wherein 락토바실러스·브레비스가 D-자일로오스의 자화성을 갖지 않는 것인 균.A bacterium in which Lactobacillus brevis does not have magnetization of D-xylose. 제 2 항에 있어서,The method of claim 2, 락토바실러스·플란타룸이 락토바실러스·플란타룸 SAM2446 주 (FERM ABP- 10438) 인 균.Lactobacillus plantarum is Lactobacillus plantarum SAM2446 strain (FERM ABP-10438). 제 3 항에 있어서,The method of claim 3, wherein 락토바실러스·브레비스가 락토바실러스·브레비스 SAM2447 주 (FERM ABP-10439) 인 균.The bacterium Lactobacillus brevis is Lactobacillus brevis SAM2447 strain (FERM ABP-10439). 제 1 항 내지 제 7 항 중 어느 한 항에 기재된 락토바실러스속균을 함유하고, 면역 조절 작용을 갖는 조성물.The composition containing the Lactobacillus genus of any one of Claims 1-7, and having an immunomodulatory effect. 제 8 항에 있어서,The method of claim 8, 상기 조성물이 음식품인 조성물.The composition is a food-drinks. 제 8 항에 있어서,The method of claim 8, 상기 조성물이 의약품인 조성물.The composition is a pharmaceutical. 제 8 항에 있어서,The method of claim 8, 상기 조성물이 사료인 조성물.The composition is a feed. 면역 조절 작용을 갖는 유산균을 분리하는 방법으로서,As a method of separating lactic acid bacteria having an immunomodulatory action, (a) 주류 제조 공정의 미생물 함유 성분으로부터 미생물을 분리하는 공정,(a) separating the microorganism from the microorganism-containing component of the liquor manufacturing process, (b) 분리된 각 미생물의 면역 조절 작용을 측정하는 공정, 및(b) measuring the immunomodulatory activity of each isolated microorganism, and (c) 면역 조절 작용이 강한 유산균을 선발하는 공정(c) A process for selecting lactic acid bacteria with strong immunomodulatory action 을 포함하는 상기 방법.The method comprising a. 제 12 항에 있어서,The method of claim 12, 유산균이 락토바실러스속균인 방법.Lactic acid bacteria is Lactobacillus genus. 제 12 항 또는 제 13 항에 있어서,The method according to claim 12 or 13, 상기 미생물 함유 성분이 발효 모로미인 방법.The microorganism-containing component is fermented moromi. 제 12 항 내지 제 14 항 중 어느 한 항에 있어서,The method according to any one of claims 12 to 14, 상기 주류 제조 공정이 와인의 양조 공정인 방법.Wherein said liquor making process is a wine brewing process.
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