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바이오패닝에 의한 Bisphenol A 친화성 펩타이드 서열의 탐색
유익근,최우석,Yoo, Ik-Keun,Choe, Woo-Seok 한국미생물학회 2013 미생물학회지 Vol.49 No.2
비스페놀 A (BPA)는 내분비계 장애물질의 하나로서 인간에게 큰 위협이 되고 있는 물질이다. 따라서 BPA의 분석 및 제거를 위해 BPA에 대해 선택적 친화성을 보이는 특정 리간드 탐색이 요구되고 있다. 본 연구에서는 초음파 처리를 동반한 바이오패닝 기법을 이용하여 파지 표면 디스플레이 라이브러리로부터 BPA에 친화성이 높은 펩타이드 서열을 탐색하였다. BPA 입자에 대한 6라운드의 positive 스크리닝과 에펜도르프 튜브 표면 재질에 대한 negative 스크리닝 과정을 실시하였고, 이를 통해 BPA에 선택적 친화성이 높은 CysLysSerLeuGluAsnSerTyrCys (CKSLENSYC) 서열을 스크리닝하였다. 또한 확보된 서열의 선택적 친화성을 검증하기 위해 BPA와 구조가 유사한 비스페놀 F (BPF), 비스페놀 S (BPS)에 대해서 교차 친화성이 있는지 평가하였고, 앞에서 선택된 서열이 BPS, BPF에 비하여 상대적으로 BPA에 대한 친화성이 높다는 것을 확인하였다. Bisphenol A (BPA) is a highly hazardous component to human since it is regarded as one of endocrine disruptors. For the analysis and/or removal of BPA, the searching for the specific ligand with a selective affinity to target BPA is required. In order to find the peptide moiety that specifically binds to BPA, the ultrasound-assisted biopanning was carried out with a phage-displayed peptide library expressing constrained heptamer. After six rounds of positive screening against BPA particles followed by the negative screening against the surface of eppendorf tube, the peptide sequence (CysLysSerLeuGluAsnSerTyrCys) with affinity to BPA was screened based on the order of frequency from the screened phage clones. To further verify the specificity of screened peptide sequence, the cross-binding affinity of the phage peptide toward BPA analogues such as Bisphenol S (BPS) and Bisphenol F (BPF) was also assessed, where the selected phage peptide showed a higher affinity to BPA over BPS and BPF.
연구논문 : Biological Aerated Filter에서 유입 질소농도에 따른 아질산화 특성
유익근 ( Ik Keun Yoo ) 한국물환경학회(구 한국수질보전학회) 2014 한국물환경학회지 Vol.30 No.1
The purpose of this study was to investigate the nitrification characteristics of biological aerated filter (BAF) packed with Ceramic media, especially focusing on nitrite build-up during nitrification. When increasing the nitrogen load above 1.63 kgNH4(+)-N/m3d, ammonium removal efficiency decreased to less than 60% and the nitrite ratio (NO2+-N/NOx-N) of higher than 75% was achieved due to the inhibitory free ammonia (FA, NH3-N) concentration and oxygen limitation. FA inhibition, however, is not recommended strategy to promote nitrite build-up since FA concentration in the reactor is coupled with decreased ammonium removal efficiency. Nitrite ratio in the effluent was also affected by aeration rate and influent ammonium concentration. Ammonium oxidation was enhanced at a higher aeration rate regardless of influent ammonium concentration but, the nitrite ratio was dependent on both aeration rate and influent ammonium concentration. While a higher nitrite ratio was obtained when BAFs were fed with 50 mgNH4(+)-N/L of influent, the nitrite ratio significantly decreased for a greater influent concentration of 200-300 mgNH4(+)-N/L. Taken together, aeration rate, influent ammonium concentration and FA concentrations kept in the BAF were found to be critical variables for nitrite accumulation in the BAF system.
Two-component System 기반 재조합균을 이용한 인공폐수에서의 중금속 인지 및 제거
홍순호,유익근,Hong, Soon-Ho,Yoo, Ik-Keun 한국미생물학회 2012 미생물학회지 Vol.48 No.3
Two component system (TCS)인 ZraS/R 및 CusS/R의 zraP와 cusC 유전자의 프로모터에 의해 green fluorescent protein (GFP)이 발현되도록 제작된 박테리아 바이오센서의 성능을 인공폐수에서 평가하였다. 제작된 박테리아 바이오센서는 실제 폐수를 모사한 인공폐수에서도 시료 중의 $Zn^{2+}$와 $Cu^{2+}$를 인지하여 GFP를 효율적으로 발현시키는 것을 확인할 수 있었다. 두 번째는 세포 표면에 금속 친화성 펩타이드가 표현되도록 제작된 구리 및 아연 제거 박테리아를 인공폐수 조건에서 평가하였다. 제작된 박테리아는 각각 ZraS/R 및 CusS/R TCS에 의해 주변의 $Zn^{2+}$와 $Cu^{2+}$를 인지하여 세포 표면에 OmpC-ZBP와 OmpC-CBP 융합 단백질을 발현시키는 시스템이다. 실험을 통해 표면에 금속 흡착 펩타이드가 발현된 재조합 균은 인공폐수 조건에서도 효과적으로 구리 및 아연을 흡착시키는 것을 확인하였다. 따라서 본 연구에서 개발된 TCS 기반 재조합 균은 인공폐수 조건에서 중금속의 인지 및 제거 기능이 효과적으로 작동하는 것이 확인되었다. Two-component system (TCS)-based bacterial zinc and copper biosensors, in which green fluorescent protein (GFP) is expressed under the control of zraP and cusC promoter in ZraS/R and CusS/R TCS, were evaluated in artificial wastewater. Bacterial biosensors developed in this study efficiently expressed GFP by the recognition of $Zn^{2+}$ and $Cu^{2+}$ in artificial wastewater. Secondly, TCS-based zinc and copper removing bacteria with the peptide displayed on cell surface were examined in artificial wastewater. Zinc and copper removing bacteria expressed the peptide as a fusion protein such as OmpC-ZBP (zinc binding peptide) and OmpC-CBP (copper binding peptide) on the cell surface when sensing exogenous $Zn^{2+}$ and $Cu^{2+}$ through ZraS/R and CusS/R TCS. The recombinant cell expressing metal-adsorbing peptide could efficiently remove copper and zinc (15 and 18 mg/g dry cell weight, respectively) in artificial wastewater. Therefore, it was demonstrated that the TCS-based recombinant cell for the recognition or removal of heavy metal functions well in artificial wastewater environment.
김동진,김건협,유익근,이규현,이태진,이용택,차기철 ( Dong Jin Kim,Gun Hyub Kim,Ik Keun Yoo,Kyu Hyun Lee,Tae Jin Lee,Yong Taek Lee,Gi Cheol Cha ) 한국물환경학회 1997 한국물환경학회지 Vol.13 No.2
A downflow biological aerated filter (BAF) packed with porous ceramic media was developed to remove the organic matter in paper industry wastewater. For about 120days, average effluent BOD, COD, and SS were stably kept to less than 20㎎/1. A reduction of 80∼90% for BOD and 75∼90% for COD was obtained at empty bed contact time (EBCT) of 1.2∼11.8 hours. There was no significant difference in removal efficiency up to the hydraulic and organic load of 40㎥/㎡day, 5㎏BOD/㎥day, respectively. The backwashing cycle at once a day was enough to maintain stable operation of BAF reactor.