본 연구는 김치 유산균을 이용한 인삼 사포닌 변환 기술 개발을 통해 ginsenoside Rg3를 대량 생산하여 건강기능성식품 소재로 이용 가능케 하는 데에 목적을 두었다. 60개의 김치 시료로부터 72종의 유산균을 분리하였고 β-glucosidase 활성을 갖는 유산균 27종을 선발하였다. 이들 균주를 대상으로 ginsenoside Rb1의 변환을 유도해본 결과 MJ-7 균주에서 ginsenoside Rg3로의 변환 활성이 관찰되었으며, 이 균주는 16S rRNA gene ...
본 연구는 김치 유산균을 이용한 인삼 사포닌 변환 기술 개발을 통해 ginsenoside Rg3를 대량 생산하여 건강기능성식품 소재로 이용 가능케 하는 데에 목적을 두었다. 60개의 김치 시료로부터 72종의 유산균을 분리하였고 β-glucosidase 활성을 갖는 유산균 27종을 선발하였다. 이들 균주를 대상으로 ginsenoside Rb1의 변환을 유도해본 결과 MJ-7 균주에서 ginsenoside Rg3로의 변환 활성이 관찰되었으며, 이 균주는 16S rRNA gene 염기서열 분석을 통한 동정 결과, Lactobacillus brevis ATCC 14869T와 99.4%의 상동성을 보임으로써 Lactobacillus brevis에 속하는 균주임이 확인되어 Lactobacillus brevis MJ-7로 명명하였다. 이 외의 분리 균주들도 16S rRNA gene 염기서열 분석을 통해 동정하였는데, 그 결과 Lactobacillus 속이 가장 많은 분포를 보였고(72%), 그 다음으로 Weissella 속이 13%, Leuconostoc 속이 12%, Pediococcus 속이 3%로 나타났다. Lactobacillus brevis MJ-7은 pH 6.5 ~ pH 7.0의 100% MRS medium에서 37℃로 배양하였을 때 최적의 생장률을 보였으며, ginsenoside Rb1과 반응시킨 결과 gypenoside ⅩⅦ, ginsenoside Rd 및 20(S)-ginsenoside Rg3로 변환되었다. 또한 cell을 pH 6.0 ~ pH 6.5의 buffer에 현탁하였을 경우와 acetone을 이용한 조효소 침전 시 뛰어난 ginsenoside Rg3로의 변환을 보였으며, acetone을 이용한 조효소 침전 시에는 pH 6.5의 buffer에서, ethanol을 이용하였을 때는 pH 6.0의 buffer에서 가장 뛰어난 효소활성을 보였다. Ginsenoside Rd 생성 균주인 MJ-28 균주와 co-culture 하여 ginsenoside Rb1의 변환을 유도할 경우 ginsenoside Rg3로의 변환이 뚜렷이 증가하였으며, acetone 침전을 이용한 반응에서도 같은 결과를 얻었다.
본 연구는 김치 유산균을 이용한 인삼 사포닌 변환 기술 개발을 통해 ginsenoside Rg3를 대량 생산하여 건강기능성식품 소재로 이용 가능케 하는 데에 목적을 두었다. 60개의 김치 시료로부터 72종의 유산균을 분리하였고 β-glucosidase 활성을 갖는 유산균 27종을 선발하였다. 이들 균주를 대상으로 ginsenoside Rb1의 변환을 유도해본 결과 MJ-7 균주에서 ginsenoside Rg3로의 변환 활성이 관찰되었으며, 이 균주는 16S rRNA gene 염기서열 분석을 통한 동정 결과, Lactobacillus brevis ATCC 14869T와 99.4%의 상동성을 보임으로써 Lactobacillus brevis에 속하는 균주임이 확인되어 Lactobacillus brevis MJ-7로 명명하였다. 이 외의 분리 균주들도 16S rRNA gene 염기서열 분석을 통해 동정하였는데, 그 결과 Lactobacillus 속이 가장 많은 분포를 보였고(72%), 그 다음으로 Weissella 속이 13%, Leuconostoc 속이 12%, Pediococcus 속이 3%로 나타났다. Lactobacillus brevis MJ-7은 pH 6.5 ~ pH 7.0의 100% MRS medium에서 37℃로 배양하였을 때 최적의 생장률을 보였으며, ginsenoside Rb1과 반응시킨 결과 gypenoside ⅩⅦ, ginsenoside Rd 및 20(S)-ginsenoside Rg3로 변환되었다. 또한 cell을 pH 6.0 ~ pH 6.5의 buffer에 현탁하였을 경우와 acetone을 이용한 조효소 침전 시 뛰어난 ginsenoside Rg3로의 변환을 보였으며, acetone을 이용한 조효소 침전 시에는 pH 6.5의 buffer에서, ethanol을 이용하였을 때는 pH 6.0의 buffer에서 가장 뛰어난 효소활성을 보였다. Ginsenoside Rd 생성 균주인 MJ-28 균주와 co-culture 하여 ginsenoside Rb1의 변환을 유도할 경우 ginsenoside Rg3로의 변환이 뚜렷이 증가하였으며, acetone 침전을 이용한 반응에서도 같은 결과를 얻었다.
Lactic acid bacteria from Kimchi samples were isolated on MRS-BPB agar plates. Among the total 72 isolated strains, 27 strains showed β-glucosidase activity. β-glucosidase activity was examined by using esculin agar plates. Ginsenoside Rb1, was treated with each of the β-glucosidase producing strain...
Lactic acid bacteria from Kimchi samples were isolated on MRS-BPB agar plates. Among the total 72 isolated strains, 27 strains showed β-glucosidase activity. β-glucosidase activity was examined by using esculin agar plates. Ginsenoside Rb1, was treated with each of the β-glucosidase producing strains. As a result, some strains were determined to be capable of hydrolyzing ginsenoside Rb1. Among these strains, a strain, designated as MJ-7, was determined to have the most potent ability to convert ginsenoside Rb1 into Rg3. The process which ginsenoside Rb1 was decomposed by strain MJ-7, was analyzed by TLC and HPLC. Among the metabolites of ginsenoside Rb1, one compound which assumed as ginsenoside Rg3 was identified by 1H-NMR and 13C-NMR spectroscopy as 20(S)-ginsenoside Rg3. 16S rRNA gene sequence analysis revealed that strain MJ-7 belongs to the genus Lactobacillus and it is closely related to Lactobacillus brevis ATCC 14869T (99.4)%, Lactobacillus hammesii DSM 16381T (98.2%) and Lactobacillus parabrevis LMG 11984T (97.5%). And seventy two isolates were found to be Lactobacillus (72%), Weissella (13%), Leuconostoc (12%) and Pediococcus (3%). Growth characteristics of strain MJ-7 were investigated to obtain basic informations for transformation of ginsenoside Rb1. The highest growth was observed at 24 hours in 100% MRS medium with pH 6.5 to 7.0 and 37℃ incubation temperature. The transformation of ginsenoside Rb1 showed different patterns by crude enzymes. Crude enzymes of strain MJ-7 precipitated by acetone retained original activity, but in case of ethanol precipitation, enzyme activity was relatively low. So, for the production of ginsenoside Rg3, the isolation of the crude enzymes precipitated by acetone is needed to increase the yield. In order to compare the effect of pH, the strain MJ-7 was mixed in 0.2 M sodium phosphate buffer (pH 3.5 to pH 8.0) and reacted ginsenoside Rb1. The maximum activities of cells, crude enzymes precipitated by acetone and ethanol were shown in pH 6.0 ~ pH 6.5, pH 6.5 and pH 6.0, respectively. When ginsenoside Rb1 was incubated with strain MJ-7 and ginsenoside Rd producing strain, the production yield of ginsenoside Rg3 was higher than when incubating with only strain MJ-7. This suggested that Rb1 was transformed through the ginsenoside Rd by sequential hydrolyses of two glucoses at C-20 of Rb1. The study demonstrates that microorganism produce specific forms of ginsenoside Rb1 may indicate that it is feasible to develop a specific biotransformation process to obtain specifically designed functional products by the appropriate combination of ginsenoside substrates and specific microbial enzymes. Therefore, it is suggested that, if ginseng extract mixed with this lactic acid bacterium, enforced amount of ginsenoside Rg3 could be produced as ginseng products. This is the first report in transformation of ginsenoside Rb1 into ginsenoside Rg3 using lactic acid bacterium, Lactobacillus brevis.
Lactic acid bacteria from Kimchi samples were isolated on MRS-BPB agar plates. Among the total 72 isolated strains, 27 strains showed β-glucosidase activity. β-glucosidase activity was examined by using esculin agar plates. Ginsenoside Rb1, was treated with each of the β-glucosidase producing strains. As a result, some strains were determined to be capable of hydrolyzing ginsenoside Rb1. Among these strains, a strain, designated as MJ-7, was determined to have the most potent ability to convert ginsenoside Rb1 into Rg3. The process which ginsenoside Rb1 was decomposed by strain MJ-7, was analyzed by TLC and HPLC. Among the metabolites of ginsenoside Rb1, one compound which assumed as ginsenoside Rg3 was identified by 1H-NMR and 13C-NMR spectroscopy as 20(S)-ginsenoside Rg3. 16S rRNA gene sequence analysis revealed that strain MJ-7 belongs to the genus Lactobacillus and it is closely related to Lactobacillus brevis ATCC 14869T (99.4)%, Lactobacillus hammesii DSM 16381T (98.2%) and Lactobacillus parabrevis LMG 11984T (97.5%). And seventy two isolates were found to be Lactobacillus (72%), Weissella (13%), Leuconostoc (12%) and Pediococcus (3%). Growth characteristics of strain MJ-7 were investigated to obtain basic informations for transformation of ginsenoside Rb1. The highest growth was observed at 24 hours in 100% MRS medium with pH 6.5 to 7.0 and 37℃ incubation temperature. The transformation of ginsenoside Rb1 showed different patterns by crude enzymes. Crude enzymes of strain MJ-7 precipitated by acetone retained original activity, but in case of ethanol precipitation, enzyme activity was relatively low. So, for the production of ginsenoside Rg3, the isolation of the crude enzymes precipitated by acetone is needed to increase the yield. In order to compare the effect of pH, the strain MJ-7 was mixed in 0.2 M sodium phosphate buffer (pH 3.5 to pH 8.0) and reacted ginsenoside Rb1. The maximum activities of cells, crude enzymes precipitated by acetone and ethanol were shown in pH 6.0 ~ pH 6.5, pH 6.5 and pH 6.0, respectively. When ginsenoside Rb1 was incubated with strain MJ-7 and ginsenoside Rd producing strain, the production yield of ginsenoside Rg3 was higher than when incubating with only strain MJ-7. This suggested that Rb1 was transformed through the ginsenoside Rd by sequential hydrolyses of two glucoses at C-20 of Rb1. The study demonstrates that microorganism produce specific forms of ginsenoside Rb1 may indicate that it is feasible to develop a specific biotransformation process to obtain specifically designed functional products by the appropriate combination of ginsenoside substrates and specific microbial enzymes. Therefore, it is suggested that, if ginseng extract mixed with this lactic acid bacterium, enforced amount of ginsenoside Rg3 could be produced as ginseng products. This is the first report in transformation of ginsenoside Rb1 into ginsenoside Rg3 using lactic acid bacterium, Lactobacillus brevis.
AI-Helper
※ AI-Helper는 부적절한 답변을 할 수 있습니다.