고 함유 메나퀴논 (menaqunone, 비타민 K2) 청국장을 개발하기 위하여 청국장으로부터 메나퀴논 생성능력이 우수한 균주를 분리•동정한 후 메나퀴논 생성을 위한 최적의 발효 특성을 시험하였다. 관능평가의 선호도와 메나퀴논 생성량이 가장 우수한 균주 BY01을 선별하였으며, 선별된 균주를 gyrB염기서열 결정법 및 생화학 분석을 통하여 Bacillus amyloliquefaciens BY01으로 명명하였다. 선발균주는 콩 추출액에서 호기적으로 배양할 경우, ...
고 함유 메나퀴논 (menaqunone, 비타민 K2) 청국장을 개발하기 위하여 청국장으로부터 메나퀴논 생성능력이 우수한 균주를 분리•동정한 후 메나퀴논 생성을 위한 최적의 발효 특성을 시험하였다. 관능평가의 선호도와 메나퀴논 생성량이 가장 우수한 균주 BY01을 선별하였으며, 선별된 균주를 gyrB염기서열 결정법 및 생화학 분석을 통하여 Bacillus amyloliquefaciens BY01으로 명명하였다. 선발균주는 콩 추출액에서 호기적으로 배양할 경우, 포자 형성율과 메나퀴논의 생성율은 반비례하였으며, 포자가 형성되기 전 6시간 동안 배양한 시험구에서 메나퀴논의 함량은4.22 μg/mL로 가장 높았다. 청국장의 본 발효에서 36시간 발효시킨 시험구의 관능평가 평점이 가장 높았다. 발효시간에 따른 메나퀴논-7및 아미노산의 생성 실험 결과, 36시간까지는 메나퀴논-7과 아미노산 (phenylalanine, tyrosine 및 tryptophan)의 생성율이 유의적으로 증가되었으나 36시간 이후부터는 메나퀴논-7의 생성율이 둔화되었으며 tryptophan > phenylalanine > tyrosine순으로 메나퀴논-7 생성을 저하시켰다. 선발된 균주에 4% (w/w)의 글리세롤을 첨가하여 43℃에서 36시간 발효시킨 청국장에서는 최고의 메나퀴논 (12.47 μg/g)이 생성되었다. 결론적으로 Bacillus amyloliquefaciens BY01로 제조된 청국장과 대조구인 상품화된 청국장 또는 낫토제품의 메나퀴논 함량을 비교할 때 메나퀴논 함량은 4배 이상 증가되었다. 따라서 B. amyloliquefaciens를 이용한 고함유 메나퀴논 청국장은 향후 칼슘흡수를 돕는 기능성 식품의 소재로 활용이 기대되는 바이다.
고 함유 메나퀴논 (menaqunone, 비타민 K2) 청국장을 개발하기 위하여 청국장으로부터 메나퀴논 생성능력이 우수한 균주를 분리•동정한 후 메나퀴논 생성을 위한 최적의 발효 특성을 시험하였다. 관능평가의 선호도와 메나퀴논 생성량이 가장 우수한 균주 BY01을 선별하였으며, 선별된 균주를 gyrB 염기서열 결정법 및 생화학 분석을 통하여 Bacillus amyloliquefaciens BY01으로 명명하였다. 선발균주는 콩 추출액에서 호기적으로 배양할 경우, 포자 형성율과 메나퀴논의 생성율은 반비례하였으며, 포자가 형성되기 전 6시간 동안 배양한 시험구에서 메나퀴논의 함량은4.22 μg/mL로 가장 높았다. 청국장의 본 발효에서 36시간 발효시킨 시험구의 관능평가 평점이 가장 높았다. 발효시간에 따른 메나퀴논-7및 아미노산의 생성 실험 결과, 36시간까지는 메나퀴논-7과 아미노산 (phenylalanine, tyrosine 및 tryptophan)의 생성율이 유의적으로 증가되었으나 36시간 이후부터는 메나퀴논-7의 생성율이 둔화되었으며 tryptophan > phenylalanine > tyrosine순으로 메나퀴논-7 생성을 저하시켰다. 선발된 균주에 4% (w/w)의 글리세롤을 첨가하여 43℃에서 36시간 발효시킨 청국장에서는 최고의 메나퀴논 (12.47 μg/g)이 생성되었다. 결론적으로 Bacillus amyloliquefaciens BY01로 제조된 청국장과 대조구인 상품화된 청국장 또는 낫토제품의 메나퀴논 함량을 비교할 때 메나퀴논 함량은 4배 이상 증가되었다. 따라서 B. amyloliquefaciens를 이용한 고함유 메나퀴논 청국장은 향후 칼슘흡수를 돕는 기능성 식품의 소재로 활용이 기대되는 바이다.
To develop a cheonggukjang with high content of menaquinone (MK), bacterial strains were isolated from the good quality cheonggukjang, and the MK productivities of the isolated strains were determined by high-performance liquid chromatography (HPLC). After sensory evaluation and MK determination, a ...
To develop a cheonggukjang with high content of menaquinone (MK), bacterial strains were isolated from the good quality cheonggukjang, and the MK productivities of the isolated strains were determined by high-performance liquid chromatography (HPLC). After sensory evaluation and MK determination, a potential probiotics strain BY01 with good sensory quality and high productivity of MK was selected. To identify the species of strain BY01, a combination of biochemical tests and gyrB sequence analysis has been carried out. The results showed that the strain BY01 was identified as Bacillus amyloliquefaciens. Preliminary study carried out in the soybean extract indicated that the fermentation under aerobic condition can produce more MK than that of anaerobic condition, and the synthesized MK mainly remained in the form of endocellular deposit. To improve the content of MK in cheonggukjang by using strain BY01, the optimal fermentation conditions were studied. The influences of pre-culture time, temperature, fermentation period, and carbon source on MK yield were analyzed. The rate of sporulation was found to inversely correlate with the MK productivity during the fermentation process. Therefore, the pre-culture should be terminated before the massive formation of spores. The changes in MK content, free amino acid amounts, and sensory index during the fermentation of cheonggukjang by B. amyloliquefaciens BY01 were investigated. The best sensory quality was appeared at 36 h of fermentation. Free amino acids and MK contents were remarkably increased during all the fermentation process. However, the increasing rate of them was slowed down after fermentation for 36 h, which may due to the accumulation of free amino acid. Three kinds of aromatic amino acids (phenylalanine, tyrosine, and tryptophan) were found to have inhibition effects on MK production during cheonggukjang fermentation, especially tryptophan, which was the most sensitive feedback inhibitor of MK production pathway, followed by phenylalanine, while tyrosine was the weakest. To increase the yield of MK in cheonggukjang, the optimum temperature was found to be 43℃, which was different from the optimum cell growth temperature (40℃). Glycerol was found to be the most effective carbon source, and the optimum concentration was 4% (w/w). The content of MK in cheonggukjang fermented with the isolated strain (B. amyloliquefaciens BY01) under the optimum condition reached as high as 12.47 μg/g, which is about four-fold higher than that of commercial cheonggukjang and natto products. This is the first report on the isolation of B. amyloliquefaciens as a superior MK producer in cheonggukjang fermentation.
To develop a cheonggukjang with high content of menaquinone (MK), bacterial strains were isolated from the good quality cheonggukjang, and the MK productivities of the isolated strains were determined by high-performance liquid chromatography (HPLC). After sensory evaluation and MK determination, a potential probiotics strain BY01 with good sensory quality and high productivity of MK was selected. To identify the species of strain BY01, a combination of biochemical tests and gyrB sequence analysis has been carried out. The results showed that the strain BY01 was identified as Bacillus amyloliquefaciens. Preliminary study carried out in the soybean extract indicated that the fermentation under aerobic condition can produce more MK than that of anaerobic condition, and the synthesized MK mainly remained in the form of endocellular deposit. To improve the content of MK in cheonggukjang by using strain BY01, the optimal fermentation conditions were studied. The influences of pre-culture time, temperature, fermentation period, and carbon source on MK yield were analyzed. The rate of sporulation was found to inversely correlate with the MK productivity during the fermentation process. Therefore, the pre-culture should be terminated before the massive formation of spores. The changes in MK content, free amino acid amounts, and sensory index during the fermentation of cheonggukjang by B. amyloliquefaciens BY01 were investigated. The best sensory quality was appeared at 36 h of fermentation. Free amino acids and MK contents were remarkably increased during all the fermentation process. However, the increasing rate of them was slowed down after fermentation for 36 h, which may due to the accumulation of free amino acid. Three kinds of aromatic amino acids (phenylalanine, tyrosine, and tryptophan) were found to have inhibition effects on MK production during cheonggukjang fermentation, especially tryptophan, which was the most sensitive feedback inhibitor of MK production pathway, followed by phenylalanine, while tyrosine was the weakest. To increase the yield of MK in cheonggukjang, the optimum temperature was found to be 43℃, which was different from the optimum cell growth temperature (40℃). Glycerol was found to be the most effective carbon source, and the optimum concentration was 4% (w/w). The content of MK in cheonggukjang fermented with the isolated strain (B. amyloliquefaciens BY01) under the optimum condition reached as high as 12.47 μg/g, which is about four-fold higher than that of commercial cheonggukjang and natto products. This is the first report on the isolation of B. amyloliquefaciens as a superior MK producer in cheonggukjang fermentation.
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