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Development of screening method for highly enriched peptides toward a multiple LPS using epoxy bead
Yun-Gon Kim(김윤곤),Chang-Soo Lee(이창수),Woo-Jae Chung(정우재),Eun-Mi Kim(김은미),Dong-Sik Shin(신동식),June-Hyung Kim(김준형),Yoon-Sik Lee(이윤식),Junho Chung(정준호),Byung-Gee Kim(김병기) 한국생물공학회 2005 한국생물공학회 학술대회 Vol.2005 No.10
전통 콩발효 식품 유래 7, 8, 4'-Trihydroxyisoflavone의 피부 생리활성 연구
박준성 ( Jun Seong Park ),김동현 ( Dong Hyun Kim ),심진섭 ( Jin-sup Shim ),김지성 ( Ji Seong Kim ),최권영 ( Kwon-young Choi ),김병기 ( Byung-gee Kim ),김덕희 ( Duck Hee Kim ),김한곤 ( Han Kon Kim ) 대한화장품학회 2010 대한화장품학회지 Vol.36 No.3
10년 이상 자연 발효된 콩 발효 식품에서 희귀 이소플라본 성분인 7, 8, 4'-trihydroxyisoflavone을 분리하였고, 분리된 성분의 항산화 효과를 비효소적인 방법으로는 DPPH 법을, 효소적인 방법으로는 xanthine oxidase법을 이용하여 확인하였다. 그 결과 7, 8, 4'-trihydroxyisoflavone는 농도 의존적으로 비타민 C와 유사한 DPPH 라디칼 소거 효능을 가지고 있음을 보여주었고 6.6 ± 0.4 μM 농도에서 xanthine oxidase에서 생성된 superoxide 라디칼을 50 % 수준까지 낮춤을 보여주었다. 또한, 이 희귀 이소플라본 성분은 UVB로 유도된 MMP-1의 생성을 초기 수준으로 억제하는 것으로 나타나 피부 노화 억제에 유용한 성분임을 확인할 수 있었다. A rare isoflavone, 7, 8, 4'-trihydroxyisoflavone, was isolated from a 10-year-old fermented soybean food. 7, 8, 4'-trihydroxyisoflavone was isolated for the first time from a Korean fermented soybean food. Evaluation tests of biological activity showed significantly inhibition activity for free radical scavenging on both non-enzymatic (DPPH system) and enzymatic method (xanthine oxidase system). DPPH radical scavenging effect of 7, 8, 4'-trihydroxyisoflavone was similar with vitamin C in a dose-dependent manner. In xanthine oxidase (XO) system 7, 8, 4'-trihydroxyisoflavone showed superoxide radical inhibition activity of 50 % at a concentration of 6.6 ± 0.4 μM. Also, the compound significantly suppressed cellular MMP-1 formation. These results suggest that 7, 8, 4'-trihydroxyisoflavone could be developed as a potential preventive or therapeutic agent against skin aging.
Tetrameric β를 이용한 고초균 포자에서의 미생물 표면 발현 모체 선별
김준형,반재구,김병기,Kim, June-Hyung,Pan, Jae-Gu,Kim, Byung-Gee 한국생물공학회 2011 KSBB Journal Vol.26 No.3
Using tetrameric ${\beta}$-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell ${\beta}$-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. ${\beta}$-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell ${\beta}$-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell ${\beta}$-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7% decrease in protease activity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.