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Fibrinolysis of Fermented Kimchi
정영기,양웅석,강정욱,공인수,김정옥,Jeong, Yong-Kee,Yang, Woong-Suk,Kang, Jeong-Ok,Kong, In-Soo,Kim, Jeong-Ok Korean Society of Life Science 1995 생명과학회지 Vol.5 No.4
Fibrinolytic activity of fermented kimchi and ingredients used for the preparation were assayed by measuring the lysis area on plasminogen-rich fibrin plate. Fermented kimchi and picked fish sauces from changlan, prwan, and anchovy showed the activity, and the activity of pickled fish sauces were high in the order of pickled changlan, picked anchovy(pickled mulchi), and pickled prawn. However, the activity of kimchi may not be attributed to pickled fish sauce because kimchi containing fish sauce did not possess activities were determined from the samples heated for 30min at 100$\circ$C. There was no changes in activities before and after heat treatment indicating the agents may be components other than protein. Since major changes occur during kimchi fermentation were increased sour taste due to production of organic acids such as lactic, citric, oxalic, and succinic acids, the authentic organic acids present were examined for fibrinolytic activities. The results indicates that the major component posses the activity is lactic acid.
이담자효모 Rhodosporidium toruloides의 막단백질 인산화와 성 Pheromone, Rhodotorucine A의 작용
정영기(Yong-Kee Jeong) 한국식품영양과학회 1997 한국식품영양과학회지 Vol.26 No.6
이담자효모 Rhodosporidium toruloides 접합형 a 세포의 막단백질 분획을 in vitro 인산화를 행한 결과 72Kd와 57Kd의 인산화 단백질이 존재하였다. 이 단백질은 접합형 A형 세포가 분비하는 sexual pheromone인 Rhodotorucine A(Rh. A)에 의하여 인산화가 저해되었다. Rh. A에 의한 인산화 저해작용은 Ca^(2+)에는 의존하였으나 Mg^(2+)이나 calmodulin에는 무관한 결과를 보였다. 인산화 저해반응에서 trigger peptidase(TPase) inhibitor인 antipain을 첨가했을 때 저해현상은 나타나지 않았다. 또한 Rh. A를 trysin으로 분해한 산물을 인산화 반응액에 첨가하였을 때 Rh. A의 효과와 같은 인산화 저해현상을 보였다. 이상의 결과로서 인산화 저해 현상은 반응액 중의 TPase와 Rh. A간의 분해산물에 의하여 나타난다고 예측할 수 있다. When the membrane protein fraction of mating type a cells of heterobasidiomycetous yeast R. toruloides was phosphorylated in vitro, two phosphorylated proteins of 72Kd and 57Kd were detected on SDS-polyacryamide gel. The phosphorylation reaction was inhibited by rhodotorucine A(Rh. A) which is a sexual pheromone secreted by mating type A cells. The inhibition of phosphorylation by Rh.A was dependent on Ca^(2+), and independent on Mg^(2+) or calmodulin. When adding trigger peptidase (TPase) inhibitor, antipain, no inhibition of phosphorylation was observed. Also, by adding the trypsin-digested product of Rh. A, the phosphorylation was inhibited as the action of Rh. A. From these results, it is expected that the inhibition of membrane protein phosphorylation should be caused by the digested product of Rh. A with TPase.
Bacillus subtilis BK-17 유래 혈전용해효소(Bacillokinase)의 고체배양
정영기(Yong-Kee Jeong),백현(Hyun Beak),서민정(Min Jeong Seo),김민정(Min Jeong Kim),이혜현(Hye Hyeon Lee),주우홍(Woo Hong Joo),김정인(Jeong In Kim),최영현(Yung Hyun Choi),정경태(Kyung Tae Chung) 한국생명과학회 2009 생명과학회지 Vol.19 No.10
곡물을 소재로 혈행개선에 효과 있는 물질을 개발하기 위하여 Bacillus subtilis BK-17을 이용하여 고체 배양을 시도하였다. 고체배양에 접종하는 미생물은 영양세포보다 포자 접종이 혈전용해효소 생산에 효과적이었다. 포자 생산을 위한 배지는 SFM (0.8% nutrient broth, 0.05% yeast extract, 10?¹ M MgCl₂, 10?³ M FeCl₃, 10?⁴ MnCl₂, 10?? M dipicolic acid, pH 6.5)이 최적이었고, 포자 생성에 대한 최적 pH 와 온도는 각각 pH6과 30℃ 였으며 포자는 배양 후 60 시간에서 가장 많은 생산을 보였다. 각종 곡물을 기질로 하였을 때 녹두가 혈전용해능이 가장 높았으며 강낭콩, 검정콩, 대두, 옥수수는 녹두의 약 90% 수준의 활성으로 높은 혈전용해능을 보였다. 반면에 보리와 밀과 같은 탄수화물 곡물은 낮은 활성을 보였다. 고체 배양한 발효 대두는 단백질과 지질 함량에 있어 발효하지 않은 대두보다 약 10% 이상의 높은 수치를 보였다. 또한 필수 아미노산과 일반 아미노산의 함량 역시 발효 대두 쪽이 약 5 - 20% 높은 것으로 보아 혈행개선기능성 뿐만 아니라 식품으로 실용 가능성이 높은 것으로 사료된다. A solid-state culture based on grain materials was attempted to produce a fibrinolytic enzyme for blood circulation improvement using Bacillus subtilis BK-17. The spore, rather than vegetative cell inoculation, of B. subtilis BK-17 on the solid-state culture was effective in the production of a fibrinolytic enzyme. Maximum spore production was obtained with a SFM medium (0.8% nutrient broth, 0.05% yeast extract, 10?¹ M MgCl₂, 10?³ M FeCl₃, 10?⁴ MnCl₂, 10?? M dipicolic acid, pH 6.5). Optimal pH and temperature were pH 6 and 30℃, respectively. The spore production reached a maximum at 60 hours of incubation. Bacillus subtilis BK-17 on the mung bean solid-state culture produced greater fibrinolytic activity, and less activity was seen in other grains such as kidney bean, soybean and corn. Protein and lipid contents of fermented soybeans were about 10 - 30% more than those of unfermented soybeans. Amino acid content was also 5 - 20% more than that of unfermented soybeans. These results indicated that fermented solid-state culture medium, fermented soybean in this case, can be utilized as a food supplement.
Purification of Internal Invertase in Rhodosporidium toruloides Mating Type A Cells
정영기(Yong-Kee Jeong),류병호(Beung-Ho Ryu) 한국식품영양과학회 1993 한국식품영양과학회지 Vol.21 No.6
이담자 효모균 Rhodosporidium toruloides의 mating type A 세포에서 세포내 invertase를 정제하였다. 세포내 invertase는 배양 세포의 파쇄액을 산침전 시킨 후 그 상등액으로부터 DEAE-Sephadex A-50, SP-Sephadex C-50 column chromatography와 Sephadex G-200 gel fitration 등의 과정을 거쳐 polyacrylamide gel disc 전기영동상 단일 효소 단백질까지 정제되었다. 정제효소의 분자량은 gel filtration에 의하여 90,000이었고, SDS-PAGE상에서는 22,000 dalttons에서 단일 band를 보여 단일종의 subunit가 4개로 구성된 단백질로 추정된다. The internal invertase was purified from cell free extract of Rhodosporidium toruloides IFO 0559-M919 by acid precipitation, ion-exchange chromatography and gel filtration to the unique enzyme protein on disc electrophoresis. We have found out that molecular weight of purified internal invertase was 90,000 by gel filtration and the purified enzyme was protein with 4 homogeneous subunits appearing as single band of 22,000daltons on SDS-polyacrylamide gel electrophoresis.