<국문초록> γ-aminobutyric acid (GABA)는 비단백성 아미노산 일종으로 L-glutamic acid의 α-탄소에 결합되어 있는 -COOH가 glutamic acid decarboxylase에 의한 탈탄산반응으로 생산된다. 인체 내에서의 GABA는 중추신경계의 억제성 ...
<국문초록> γ-aminobutyric acid (GABA)는 비단백성 아미노산 일종으로 L-glutamic acid의 α-탄소에 결합되어 있는 -COOH가 glutamic acid decarboxylase에 의한 탈탄산반응으로 생산된다. 인체 내에서의 GABA는 중추신경계의 억제성 신경전달물질로 작용하고, 혈압 상승억제, 식욕과 포만감등에 관여하여 체중 감소를 야기한다는 보고가 있다. 처음에 GABA는 포유동물의 뇌 조직에서 발견되었다. 식품중에서는 특히 한국의 전통발효식품인 김치가 고동도의 GABA를 생산한다고 보고되었다. 이 연구의 목적은 김치로부터 GABA를 생산할 수 있는 유산균을 분리하고, GABA 생산수율이 뛰어난 균주를 동정하기 위함이다. 김치로부터 유산균 324중을 분리하여 1%의 sodium L-glutamate (MSG)가 첨가된 MRS배지에 접종하고 30℃에서 36시간 배양하였다. 이 중 7균주는 Thin Layer Chromatography (TLC) 분석에 의해 GABA 생산이 확인되었다. 이들 균주의 GABA 전환율은 1%의 MSG가 첨가한 경우가 가장 높았다. 그러나 GABA의 가장 높은 수확량은 5%의 MSG가 첨가된 MRS배지에서 나타냈다. 7균주중 GABA 생산능력이 탁원했던 CK12, CC178, CKK215 및 CKK222는 API과 16S rDNA 분석에 의해 Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus brevis CKK215 (ATCC14869T) 및 Lactobacillus brevis CKK222 (ATCC14869T)로 동정되었다. 7균주에 의해 생산된 GABA를 정량 측정하고, 정확한 GABA 수율을 구하기 위해 High Pressure Liquid Chromatography (HPLC)를 사용하였다. 1% MSG 와 5% MSG를 첨가하여 24시간, 48시간 그리고 72시간 균주를 배양하였다. 1% MSG 첨가배지의 경우 48시간 배양에서는 95%이상 GABA로 전환시킬 수 있었고 이중 5균주는 GABA 수율이 100%였다. 72시간 배양에서는 7균주 모두 100% GABA로 전환하였다. 5% MSG 첨가배지의 경우 48시간 배양에서는 50%이상 GABA로 전환시킬 수 있었고 이중 Lactobacillus CKK222의 GABA 생산량이 94.87%로 가장 높았다. 72시간배양에서는 Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus sp. CK38, Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus sp. CC325, Lactobacillus sp. CKK214, Lactobacillus brevis CKK215 (ATCC14869T) 및 Lactobacillus brevis CKK222 (ATCC14869T)의 수율이 각각 86.21%, 78.34%, 87.84%, 90.03%, 90.92%, 98.22 및 100%였다. MSG첨가배지에 배양 후 pH는 MSG를 넣지 않은 MRS배지보다 더 높았다. 이는 식품의 발효과정중에 생산되는 유산균의 신맛을 경감시키는 것으로 보인다.
<국문초록> γ-aminobutyric acid (GABA)는 비단백성 아미노산 일종으로 L-glutamic acid의 α-탄소에 결합되어 있는 -COOH가 glutamic acid decarboxylase에 의한 탈탄산반응으로 생산된다. 인체 내에서의 GABA는 중추신경계의 억제성 신경전달물질로 작용하고, 혈압 상승억제, 식욕과 포만감등에 관여하여 체중 감소를 야기한다는 보고가 있다. 처음에 GABA는 포유동물의 뇌 조직에서 발견되었다. 식품중에서는 특히 한국의 전통발효식품인 김치가 고동도의 GABA를 생산한다고 보고되었다. 이 연구의 목적은 김치로부터 GABA를 생산할 수 있는 유산균을 분리하고, GABA 생산수율이 뛰어난 균주를 동정하기 위함이다. 김치로부터 유산균 324중을 분리하여 1%의 sodium L-glutamate (MSG)가 첨가된 MRS배지에 접종하고 30℃에서 36시간 배양하였다. 이 중 7균주는 Thin Layer Chromatography (TLC) 분석에 의해 GABA 생산이 확인되었다. 이들 균주의 GABA 전환율은 1%의 MSG가 첨가한 경우가 가장 높았다. 그러나 GABA의 가장 높은 수확량은 5%의 MSG가 첨가된 MRS배지에서 나타냈다. 7균주중 GABA 생산능력이 탁원했던 CK12, CC178, CKK215 및 CKK222는 API과 16S rDNA 분석에 의해 Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus brevis CKK215 (ATCC14869T) 및 Lactobacillus brevis CKK222 (ATCC14869T)로 동정되었다. 7균주에 의해 생산된 GABA를 정량 측정하고, 정확한 GABA 수율을 구하기 위해 High Pressure Liquid Chromatography (HPLC)를 사용하였다. 1% MSG 와 5% MSG를 첨가하여 24시간, 48시간 그리고 72시간 균주를 배양하였다. 1% MSG 첨가배지의 경우 48시간 배양에서는 95%이상 GABA로 전환시킬 수 있었고 이중 5균주는 GABA 수율이 100%였다. 72시간 배양에서는 7균주 모두 100% GABA로 전환하였다. 5% MSG 첨가배지의 경우 48시간 배양에서는 50%이상 GABA로 전환시킬 수 있었고 이중 Lactobacillus CKK222의 GABA 생산량이 94.87%로 가장 높았다. 72시간배양에서는 Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus sp. CK38, Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus sp. CC325, Lactobacillus sp. CKK214, Lactobacillus brevis CKK215 (ATCC14869T) 및 Lactobacillus brevis CKK222 (ATCC14869T)의 수율이 각각 86.21%, 78.34%, 87.84%, 90.03%, 90.92%, 98.22 및 100%였다. MSG첨가배지에 배양 후 pH는 MSG를 넣지 않은 MRS배지보다 더 높았다. 이는 식품의 발효과정중에 생산되는 유산균의 신맛을 경감시키는 것으로 보인다.
γ-aminobutyric acid (GABA) is a ubiquitous non-protein amino acid produced primarily by the glutamic acid decarboxylase (GAD). GABA function as a major inhibitory neurotransmitter in animals, and is involved in the regulation of cardiovascular functions, such as blood pressure and heart rate, and p...
γ-aminobutyric acid (GABA) is a ubiquitous non-protein amino acid produced primarily by the glutamic acid decarboxylase (GAD). GABA function as a major inhibitory neurotransmitter in animals, and is involved in the regulation of cardiovascular functions, such as blood pressure and heart rate, and play a role in the sensations of pain and anxiety. GABA was discovered in brain tissues of mammals. It was reported some other sources of foods have high concentration of GABA, containing kimchi, a traditional fermented food in Korea. The purpose of this study is to select the lactic acid bacteria (LAB) that can produce GABA from kimchi, to identify the GABA producing positive strains and to analyze the yield of GABA. 324 strains of lactic acid bacteria (LAB) were isolated from kimchi and incubated in the MRS medium added 1% sodium L-glutamate (MSG) at 30℃ for 36 hours. 7 strains of LAB were determined putatively as GABA producing LAB, and it was confirmed by Thin Layer Chromatography (TLC) analysis. It was considered that the conversion ratio in MRS medium with 1% MSG was the highest. However, the highest yield of GABA was shown in the MRS medium added 5% MSG. The strains named as CK12, CC178, CKK215 and CKK222 were identified by API test and 16S rDNA analysis as Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus brevis CKK215 (ATCC14869T) and Lactobacillus brevis CKK222 (ATCC14869T), respectively. The quantitative analysis of GABA produced by LAB was measured by High Pressure Liquid Chromatography (HPLC). The strains were incubated for 24 hours, 48 hours and 72 hours at 30℃ in the MRS medium added 1% or 5% MSG. The results of HPLC showed that more than 95% MSG could be converted to GABA within 48 hours in culture broth that added 1% MSG by 7 strains. Especially, the conversion ratio of 5 strains was 100%. The conversion ratio of 7 strains was all reached to 100% within 72 hours. More than 50% MSG could be converted to GABA within 48 hours in culture added 5% MSG by 7 strains, and the conversion ratio of Lactobacillus brevis CKK222 (ATCC14869T) was high as 94.87%, therefore, Lactobacillus brevis CKK222 (ATCC14869T) was considered as the most promising strain in GABA production among 7 GABA producing strains. For the culture of 72 hours the conversion ratio of Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus sp. CK38, Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus sp. CC325, Lactobacillus sp. CKK214, Lactobacillus brevis CKK215 (ATCC14869T) and Lactobacillus brevis CKK222 (ATCC14869T) were shown as 86.21%, 78.34%, 87.84%, 90.03%, 90.92%, 98.22% and 100%, respectively. pH of the supernatant after incubation in MRS medium with MSG was higher than in just MRS medium. It was considered that the sour taste might be alleviated in case of LAB fermentation during the food processing.
γ-aminobutyric acid (GABA) is a ubiquitous non-protein amino acid produced primarily by the glutamic acid decarboxylase (GAD). GABA function as a major inhibitory neurotransmitter in animals, and is involved in the regulation of cardiovascular functions, such as blood pressure and heart rate, and play a role in the sensations of pain and anxiety. GABA was discovered in brain tissues of mammals. It was reported some other sources of foods have high concentration of GABA, containing kimchi, a traditional fermented food in Korea. The purpose of this study is to select the lactic acid bacteria (LAB) that can produce GABA from kimchi, to identify the GABA producing positive strains and to analyze the yield of GABA. 324 strains of lactic acid bacteria (LAB) were isolated from kimchi and incubated in the MRS medium added 1% sodium L-glutamate (MSG) at 30℃ for 36 hours. 7 strains of LAB were determined putatively as GABA producing LAB, and it was confirmed by Thin Layer Chromatography (TLC) analysis. It was considered that the conversion ratio in MRS medium with 1% MSG was the highest. However, the highest yield of GABA was shown in the MRS medium added 5% MSG. The strains named as CK12, CC178, CKK215 and CKK222 were identified by API test and 16S rDNA analysis as Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus brevis CKK215 (ATCC14869T) and Lactobacillus brevis CKK222 (ATCC14869T), respectively. The quantitative analysis of GABA produced by LAB was measured by High Pressure Liquid Chromatography (HPLC). The strains were incubated for 24 hours, 48 hours and 72 hours at 30℃ in the MRS medium added 1% or 5% MSG. The results of HPLC showed that more than 95% MSG could be converted to GABA within 48 hours in culture broth that added 1% MSG by 7 strains. Especially, the conversion ratio of 5 strains was 100%. The conversion ratio of 7 strains was all reached to 100% within 72 hours. More than 50% MSG could be converted to GABA within 48 hours in culture added 5% MSG by 7 strains, and the conversion ratio of Lactobacillus brevis CKK222 (ATCC14869T) was high as 94.87%, therefore, Lactobacillus brevis CKK222 (ATCC14869T) was considered as the most promising strain in GABA production among 7 GABA producing strains. For the culture of 72 hours the conversion ratio of Lactobacillus pentosus CK12 (JCM 1558T), Lactobacillus sp. CK38, Lactobacillus brevis CC178 (ATCC14869T), Lactobacillus sp. CC325, Lactobacillus sp. CKK214, Lactobacillus brevis CKK215 (ATCC14869T) and Lactobacillus brevis CKK222 (ATCC14869T) were shown as 86.21%, 78.34%, 87.84%, 90.03%, 90.92%, 98.22% and 100%, respectively. pH of the supernatant after incubation in MRS medium with MSG was higher than in just MRS medium. It was considered that the sour taste might be alleviated in case of LAB fermentation during the food processing.
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