KR20070053858A - Multi-starter set of lactic acid bacteria for increasing s-adenosylmethionine in fermented food - Google Patents
Multi-starter set of lactic acid bacteria for increasing s-adenosylmethionine in fermented food Download PDFInfo
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Abstract
본 발명은 우울증, 관절염, 간질환 등에 효과가 있는 S-adenosylmethionine(SAM)의 생성이 우수한 젖산균주를 선정하여 발효식품 종균으로 사용하는 것이다. 상세하게는 젖산균의 SAM 생성량을 속, 종별로 조사하여 생성 우수종균을 선별하여 종균 set으로 발효식품에 응용하는 단계로 구성되어 있다.The present invention is to select the lactic acid strain excellent in the production of S-adenosylmethionine (SAM) that is effective in depression, arthritis, liver disease, etc. to use as a fermented food spawn. In detail, it is composed of the step of selecting the excellent production spawn by applying the production of SAM in the lactic acid bacteria by genus, species, and applying to the fermented food as a seed set.
본 발명은 젖산균 특성에 따라 김치를 비롯한 젖산 발효 식품에 응용하여 식품내에 SAM 생성량을 증진시키는 데 있다. The present invention is to improve the amount of SAM produced in foods by applying to lactic acid fermented foods, including kimchi according to the characteristics of lactic acid bacteria.
젖산균, S-adenosylmethionine, 젖산발효식품 Lactic Acid Bacteria, S-adenosylmethionine, Lactic Acid Fermented Food
Description
도 1은 균주분양기관으로부터 분양받은 젖산균으로부터 SAM 생성량이 우수한 균주를 선별하여 젖산균 종균 set을 제조하여 식품에 적용한 절차도Figure 1 is a procedure of applying the food by producing a lactic acid bacteria spawn set by selecting strains excellent in the amount of SAM production from the lactic acid bacteria received from the strain culture institution
도 2는 발효된 김치로부터 김치젖산균을 분리하여 SAM 생성량이 우수한 균주를 선별하여 젖산균 종균 set을 제조하여 식품에 적용한 절차도 Figure 2 is a procedure of separating the kimchi lactic acid bacteria from fermented kimchi to select strains excellent in the amount of SAM production to produce a lactic acid bacterium seed set and applied to food
본 발명은 젖산균 중에서 우울증, 류머티스 관절염, 간질환 등에 효과가 있는 S-adenosylmethionine(SAM) 생성이 우수한 균주를 선정하여 발효식품 종균으로 사용하는 것이다.The present invention is to select a strain excellent in the production of S-adenosylmethionine (SAM) that is effective in depression, rheumatoid arthritis, liver disease, etc. among the lactic acid bacteria to be used as fermented food spawn.
최근에 기능성을 증진시켜서 젖산균을 종균으로 사용한 식품은 요구르트가 대표적이며 헬리코박터 파이로리를 억제하여 위를 보호하는 것, 알콜흡수를 억제하여 간을 보호하는 것, 장내 유해효소의 생산을 억제하고 간독성 물질로부터 간을 보호하는 역할을 하는 것 등이 있다. 또한, 유해세균 억지력이 있는 김치 젖산균을 이용하여 요구르트 종균으로 사용한 것이 있었으며 SAM 생성력이 증진된 젖산균을 이용한 것은 없었다. In recent years, foods that use lactic acid bacteria as a spawn seed to enhance functionality are typical of yogurt, which protects the stomach by inhibiting Helicobacter pylori, protects the liver by inhibiting alcohol absorption, inhibits the production of intestinal harmful enzymes, It also plays a role in protecting the liver. In addition, some of them were used as yogurt spawns using kimchi lactic acid bacteria having harmful bacterium inhibitory ability, and none of them had enhanced SAM production ability.
본 발명은 균주 분양기관으로부터 식품에서 흔히 발견되는 젖산균을 분양받아서 SAM 생성량을 조사한 결과, 종간(Species level) 및 속간(Genus level) 젖산균에 따라 생성량이 차이지는 것으로 나타나, SAM을 다량생성하는 균주 들을 구분하여 젖산균 종균 set을 제조하였다. 또한, 김치의 천연균총으로부터 SAM을 다량생성하는 균주 들을 구분하여 젖산균 종균 set을 제조하였다. According to the present invention, the amount of production of SAM is determined by receiving lactic acid bacteria commonly found in foods from a strain distribution institution, and the production amount is different depending on the species level and genus level lactic acid bacteria. The lactic acid bacteria spawn set was prepared separately. In addition, lactic acid bacteria spawn set was prepared by distinguishing the strains that produce a large amount of SAM from kimchi natural flora.
본 발명은 젖산균 중에서 S-adenosylmethionine(SAM) 생성이 우수한 균주를 선정하여 발효식품 종균 또는 종균 set으로 사용하는 것이다.The present invention is to select a strain excellent in the production of S-adenosylmethionine (SAM) among the lactic acid bacteria to use as a fermented food spawn or spawn set.
실험에 사용된 젖산균은 한국생명공학연구원과 한국식품연구원에서 분양받은 51균주(표 1)와 김치에서 분리한 180균주(표 2)를 사용하였으며 사용 김치 및 김치제조는 다음과 같다.The lactic acid bacteria used in the experiment were 51 strains (Table 1) distributed from Korea Research Institute of Biotechnology and Korea Food Research Institute and 180 strains separated from Kimchi (Table 2). Kimchi and Kimchi production were as follows.
실험에 사용한 김치는 구매하였거나 제조하였으며 제조김치의 원부재료는 절임배추 100g당 파 1.8g, 고춧가루 2.5g, 마늘 3g, 생강 0.5g, 젓갈 3g이었다. Kimchi used in the experiment was purchased or manufactured. The raw ingredients of the prepared kimchi were 1.8g of green onions per 100g of pickled cabbage, 2.5g of red pepper powder, 2.5g of garlic, 0.5g of ginger, and 3g of salted fish.
김치는 멸치젓을 넣어서 5℃와 10℃에서 발효 시킨 것, 새우젓, 까나리액젓, 젓갈 무첨가하여 10℃에서 발효시킨 것, 시중에서 구매한 것으로부터 김치의 맛이 가장 좋은 적숙기(pH4.2, 총산도 0.6 ∼ 0.8%)일때 젖산균 선택배지를 이용하여 분 리하였으며 API 50 CHL system으로 동정하였다. Kimchi fermented at 5 ℃ and 10 ℃ with anchovy marinated, fermented at 10 ℃ with no salted shrimp, canary aquifer, salted fish, and commercially prepared kimchi (pH4.2, total) When the acidity was 0.6 ~ 0.8%), it was isolated using lactic acid bacteria selection medium and identified by API 50 CHL system.
분리된 젖산균은 MRS agar 위에 반투막을 놓고 그 위에 성장시킨 후에 미생물 균체를 1 M, formic acid, 10ml로 현탁하여 회수하였고 동결건조 하였으며 증류수, 1ml에 녹여서 HPLC로 분석하였다. The isolated lactic acid bacteria were recovered by placing a semipermeable membrane on MRS agar, suspended in 1 M, formic acid, and 10 ml, lyophilized, and dissolved in distilled water and 1 ml.
KCTC: 한국생명공학연구원 균주, KFRI, AP: 한국식품연구원 균주KCTC: Korea Biotechnology Research Institute strain, KFRI, AP: Korea Food Research Institute strains
각 김치에서 분리된 경시적 우세 젖산균과 분양된 표준 균주의 세포내의 SAM을 분석하기 위하여 배양하여 세포를 1M Formic acid를 사용하여 회수하였고 HPLC(Jasco, japan)를 이용하여 분석하였다. HPLC에 사용한 column은 CAPCELL PAK C18을 사용하였으며, 일정한 농도로 제조한 sample을 20㎕ 주입하여 0.25M ammonium acetate와 100% methanol을 buffer로 사용하여 분석하였다. 이때의 HPLC의 분석 조건은 표 3과 같다.The cells were harvested using 1M Formic acid and analyzed by HPLC (Jasco, Japan) to analyze SAMs in cells of predominant lactic acid bacteria and standard strains isolated from each kimchi. For the column used for HPLC, CAPCELL PAK C18 was used, and 20 µl of a sample prepared at a constant concentration was injected and analyzed using 0.25M ammonium acetate and 100% methanol as a buffer. HPLC analysis conditions at this time are shown in Table 3.
실험에 사용된 분양된 젖산구균(Enterococcus)의 SAM 생성량은 평균 0.63 mM로 나타났다(표 4). The mean amount of SAM produced in the cultured Lactobacillus ( Enterococcus ) used in the experiment was 0.63 mM (Table 4).
젖산간균(Lactobacillus) 속의 SAM 함량을 살펴보면, 각 분양된 표준 균주의 속을 비교했을 경우, 평균 SAM 함량이 0.94mM로 나타나 두 번째로 높았다(표 5, 6). 이 속에서는 Lactobacillus delbrueckii subsp. delbrueckii 균주가 가장 높은 1.33mM의 SAM함량을 가지고 있는 것으로 나타났으며 다음으로 L. pentosus, L. curvatus, L. collinoides 균주 순으로 높은 SAM 생성을 나타냈다(표 5) Lactobacillus Looking at the SAM content of the genus, when comparing the genus of each standard strain, the average SAM content was 0.94mM was the second highest (Tables 5 and 6). In this Lactobacillus delbrueckii subsp. delbrueckii The strain had the highest SAM content of 1.33 mM , followed by L. pentosus , L. curvatus, L. collinoides High SAM production in order of strain (Table 5)
Leuconostoc 속의 SAM 생성량은 평균 1.16mM로 나타나, Lactobacillus속, Lactococcus 속, Pediococcus 속, Enterococcus 속, Weissella 속, 기타(Streptococcus 속, Aerococcus 속)보다 균주 내 SAM 함량이 월등히 높았다. 특히 Leuconostoc 속에서도 Leuconostoc mesenteroides subsp. mesenteroides의 SAM함량이 1.29mM로 나타나 가장 높았다(표 6). Leuconostoc SAM production in the genus is 1.16mM on average, Lactobacillus genus, Lactococcus genus, Pediococcus genus, Enterococcus The contents of SAM in the strain were much higher than those of the genus, Weissella , and others ( Streptococcus , Aerococcus ). Especially in the Leuconostoc Leuconostoc mesenteroides subsp. The SAM content of mesenteroides was 1.29 mM, which was the highest (Table 6).
Pediococcus 속의 균주 내 SAM 함량이 평균 0.73으로 나타나, 표준 균주 내에서 세 번째로 SAM함량이 높은 것으로 나타났으며, P. pentosacius가 0.92mM로 나타나 이중에서 가장 높았다.(표 7). Pediococcus The mean SAM content of the genus strain was 0.73, which was the third highest SAM content in the standard strain, and P. pentosacius was 0.92 mM, the highest among them (Table 7).
Weissella속은 평균이 0,51mM로 나타나 생성량이 낮게 나타났다(표 8). The genus Weissella showed a low production with an average of 0,51 mM (Table 8).
표 7. 분양된 Pediococcus의 SAM 생성량Table 7. SAM Production of Pediococcus Sold
Weissella속은 평균이 0,51mM로 나타나 생성량이 낮게 나타났다(표 8). The genus Weissella showed a low production with an average of 0,51 mM (Table 8).
기타 Streptococcus 속, Aerococcus 속 , Lactococcus 속의 균주내 SAM 함량을 살펴보면 다른 균주들에 비해 SAM 함량이 매우 낮은 것으로 나타났고, Lactococcus 속, Aerococcus 속, Streptococcus 속의 순으로 나타났다(표 9).Other Streptococcus genus, Aerococcus Genus , Lactococcus The contents of SAM in the genus were found to be very low in SAM compared to other strains. Lactococcus , Aerococcus , Streptococcus In order of genus (Table 9).
따라서, 젖산균 속 수준에서는 Leuconostoc이, 다음으로 Lactobacillus가 우수하였고 종 수준에서는 Lactobacillus delbrueckii subsp. delbrueckii , Leuconostoc mesenteroides subsp. mesenteroides , L. pentosus , Leu . mesenteroides subsp. dextranicum , L. fructovorans , L. cuvatus , L. collinoides, Leu . gelidum 등 순으로 우수하였다(표 5-9).Therefore, the lactic acid level in the Leuconostoc, Lactobacillus was an excellent servant to the next level in the Lactobacillus delbrueckii subsp. delbrueckii , Leuconostoc mesenteroides subsp. mesenteroides , L. pentosus , Leu . mesenteroides subsp. dextranicum , L. fructovorans , L. cuvatus , L. collinoides, Leu . gelidum Excellent in equal order (Table 5-9).
멸치젓을 넣어서 5℃에서 발효시킨 김치에서 분리한 젖산균의 SAM 생성량은 L.. bifermentans QMW327 균주가 1.43mM을 생성하여 가장 높았다(표 10)The amount of SAM produced by lactic acid bacteria isolated from Kimchi fermented at 5 ℃ by fermentation of anchovy was measured in L .. bifermentans The QMW327 strain produced the highest 1.43 mM (Table 10)
멸치젓을 넣어서 10℃에서 발효시킨 김치에서 분리한 젖산균의 SAM 생성량은 L.. curvatus SKP313 균주가 1.22mM을 생성하여 가장 높았고 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) SMY329가 다음으로 높았다(표 11)The amount of SAM produced by lactic acid bacteria isolated from Kimchi fermented at 10 ℃ by fermentation of anchovy was L .. curvatus Leu to the highest SKP313 strains produce a 1.22mM. mesenteroides subsp. . mesenteoides (or dextranicum ) SMY329 was next highest (Table 11)
새우젓을 넣어서 10℃에서 발효시킨 김치에서 분리한 젖산균의 SAM 생성량은 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) KSK417 균주가 1.54mM을 생성하여 가장 높았고 동종의 KSP426이 다음으로 높았다(표 12)The amount of SAM produced by lactic acid bacteria isolated from kimchi fermented at 10 ℃ with shrimp paste was Leu . mesenteroides subsp. . mesenteoides (or dextranicum ) The KSK417 strain produced 1.54mM, the highest, and the homologous KSP426 was the next highest (Table 12).
까나리 액젓을 넣어서 10℃에서 발효시킨 김치에서 분리한 젖산균의 SAM 생성량은 L.. plantarum KKM408 균주가 1.43mM을 생성하여 가장 높았고 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) SKKP 422가 다음으로 높았다(표 13)The amount of SAM produced by lactic acid bacteria isolated from Kimchi fermented at 10 ℃ by adding Canary fish sauce was L .. plantarum The KKM408 strain produced 1.43 mM and was the highest and Leu . mesenteroides subsp. . mesenteoides (or dextranicum ) SKKP 422 was next highest (Table 13)
젓갈을 넣지 않고 10℃에서 발효시킨 김치에서 분리한 젖산균의 SAM 생성량은 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) KFM 402 균주가 1.44mM을 생성하여 가장 높았고 L.. brevis KFP422가 다음으로 높았다(표 14)The amount of SAM produced by lactic acid bacteria isolated from Kimchi fermented at 10 ℃ without adding salted fish was Leu . mesenteroides subsp. . mesenteoides (or dextranicum ) KFM 402 strain produced 1.44 mM, the highest L .. brevis KFP422 was next highest (Table 14)
시판중인 J 김치에서 분리한 젖산균의 SAM 생성량은 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) KJM 401 균주가 1.51mM을 생성하여 가장 높았고 L.. plantarum KJM405가 다음으로 높았다(표 15)The production of SAM of lactic acid bacteria isolated from commercial J kimchi was Leu . mesenteroides subsp. . mesenteoides (or dextranicum ) KJM 401 strain produced 1.51 mM and was the highest in L .. plantarum KJM405 was next highest (Table 15)
따라서, SAM생성량은 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) KSK417 균주가 1.54mM을 생성하여 가장 높았고 같은 종의 균주 들이 대부분의 김치에서 생성량이 많았다. Lactobacillus에서는 L. plantarum, L. bifermentans, L. brevis 등의 높았다(표 10-15).Thus, the amount of SAM produced is Leu . mesenteroides subsp. . Mesenteoides (or dextranicum ) KSK417 produced 1.54mM, which was the highest, and most of the same species were produced in most kimchi. In Lactobacillus, L. plantarum, L. bifermentans and L. brevis were high (Table 10-15).
[실시예]EXAMPLE
SAM 생성량이 높은 균주를 4균주씩 동시에 인공김치배지에 첨가하여 10℃에 10일간 정치배양하였고 HPLC로 분석하였다. 인공김치는 상기김치 조성표에 물을 김치 무게비에 2배량 넣어서 원 김치 포함하여 총 3배로 하였으며 염도를 2.5%로 재조정하였다. 이것을 워링블렌더 믹서로 갈고 4℃에서 1시간 진탕하여 원부재료 안에 있는 물질을 용출 시켰으며 저속 원심한 후에 상등액을 인공김치용 배양액으로 하였다. 접종 젖산균은 MRS broth에 2차 계대 배양한 후에 원심분리하였고 인공김치 배양액으로 재현탁하여 0.1%(v/v)되게끔 SAM 생성 우수균을 4 균주씩 접종하였다. 인공김치 배양액은 배양후에 100ml를 원심분리하여 젖산균체를 얻었으며 10ml, 1M formic acid를 처리하여 동결건조 한 후에 HPLC로 분석하였다. 그 결과 김치에서 분리한 젖산균 혼합 종균이 SAM을 1.7mM을 생성하여 가장 많았고 무접종구보다 1.84배 많았으며 Leuconostoc 혼합종균, Lactobacillus 혼합종균 순으로 높았다.Strains with high SAM production were added to artificial kimchi medium at the same time by 4 strains and left to culture at 10 ° C. for 10 days and analyzed by HPLC. Artificial kimchi was added to the kimchi composition table twice the amount of water in the kimchi weight ratio, including three times the total kimchi, and adjusted the salinity to 2.5%. This was ground with a blending mixer and shaken at 4 ° C. for 1 hour to elute the substances in the raw materials. The supernatant was used as an artificial kimchi culture medium after low speed centrifugation. Inoculation Lactobacillus was centrifuged after secondary passage in MRS broth and resuspended in artificial kimchi culture to inoculate 4 strains of SAM-producing bacteria to 0.1% (v / v). After the incubation, the artificial kimchi culture was centrifuged to obtain lactic acid bacteria, and 10ml, 1M formic acid was lyophilized and analyzed by HPLC. The result was a mixed inoculum of lactic acid bacteria isolated from Kimchi most common non-vaccinated guboda 1.84 times many of SAM to create a 1.7mM Leuconostoc It was mixed with seed, Lactobacillus mixed seed order.
표. 인공김치액에서 10일 배양시킨 젖산균체 내의 SAM 함량table. SAM Content in Lactic Acid Cells Cultured in Artificial Kimchi for 10 Days
1) Lactobacillus 분양균 mixture set: Lactobacillus delbrueckii subsp. delbrueckii KCTC 1047, Lactobacillus pentosus KFRI 481, L. curvatus KFRI 654, L. collinoides KFRI 11821) Lactobacillus predator mixture set: Lactobacillus delbrueckii subsp. delbrueckii KCTC 1047, Lactobacillus pentosus KFRI 481, L. curvatus KFRI 654, L. collinoides KFRI 1182
2) Leuconostoc 분양균 mixture set: Leuconostoc mesenteroides subsp. mesenteroides KCTC 3100, Leu . mesenteroides subsp. dextranicum KCTC 3530, Leuconostoc gelidum KCTC 3527, Leuconostoc citreum KCTC 35262) Leuconostoc Breeding mixture mixture set: Leuconostoc mesenteroides subsp. mesenteroides KCTC 3100, Leu . mesenteroides subsp. dextranicum KCTC 3530, Leuconostoc gelidum KCTC 3527, Leuconostoc citreum KCTC 3526
3) 김치 분리 젖산균 mixture set(다양한 김치에서 SAM 생성량이 우수한 젖산균유래 혼합종균): 시판김치유래균 Leu . mesenteroides subsp.. mesenteoides(또는 dextranicum) KJM 401, 까나리액젓 첨가구 L.. plantarum KKM408, 5℃ 발효 멸치젓 첨가구 L.. bifermentans QMW327, 10℃ 발효 멸치젓 첨가구 L.. curvatus SKP313 3) Kimchi Isolation Lactic Acid Bacteria mixture set (Lactic acid bacteria-derived mixed species having excellent SAM production in various kimchi): Commercially available Kimchi-derived Leu . mesenteroides subsp. . mesenteoides (or dextranicum ) KJM 401, Canary fish sauce added L .. plantarum KKM408, 5 ℃ Fermented Anchovy Sauce L .. bifermentans QMW327, 10 ℃ Fermented Anchovy Sauce L .. curvatus SKP313
본 발명에 의해 발효식품내에 접종된 젖산균에 의하여 균체내의 SAM 생성량을 증가시킬 수 있었다.According to the present invention, the amount of SAM produced in the cells could be increased by the lactic acid bacteria inoculated into the fermented food.
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KR100942290B1 (en) * | 2007-10-25 | 2010-02-16 | 두두원발효(주) | Anti hypochondria composition with soy yogurt fermented by kimchi lactic acid bacteria |
WO2011040820A1 (en) * | 2009-09-30 | 2011-04-07 | Fjell & Fjord Mat As | Lactic acid bacteria suitable for producing partly fermented meat and a process for producing partly fermented meat by establishing conditions suitable for the proliferation of such lactic acid bacteria |
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