• 생명과학회지
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• 제22권8호
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• pp.1009-1017
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• 2012

Rodobacter sphaeroides에서 orf282 유전자는 cbb3 terminal oxidase를 암호화하는 ccoNOQP 오페론과 혐기적 활성자인 FnrL을 암호화하는 fnrL 유전자 사이에 있으며, 아직은 기능이 잘 알려지지 않았다. orf282 유전자의 기능을 알기 위해 우리는 orf282의 일부를 삭제함으로써 유전자를 붕괴시켜 orf282-minus mutant를 제조하였다. 두개의 FnrL 결합 부위가 orf282의 upstream에 존재한다는 것이 밝혀져 있으며, orf282 유전자가 FnrL에 의해 양성적으로 조절된다는 것이 증명되었다. orf282 유전자는 B875와 B800-850 spectral complexes의 형성과 관련이 없다. orf282 mutant에서의 cbb3 oxidase 활성을 wild type와 비교해보면 orf282 유전자가 ccoNOQP 오페론의 조절과 cbb3 cytochrome c oxidase의 생합성과 무관하다는 것을 알 수 있다. orf282 mutant의 구조 유전자인 nifH와 조절유전자인 nifA의 프로모터 활성이 증가한 것은 orf282 유전자 산물이 nifH와 nifA의 발현에서 음성적 effector로 작용한다는 것을 시사한다.

• BMB Reports
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• 제29권1호
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• pp.88-91
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• 1996

The anaerobically expressed gene aeg-46.5, which had been identified by the operon fusion technique with a hybrid bacteriophage of ${\lambda}$ and Mu, ${\lambda}$placMu53, was studied for its expression pattern and growth. The expression of aeg-46.5 was studied in the wild-type cell and mutant cells that have mutation (s) in the control gene of anaerobic respiration (fnr) and nitrate response (narL and narP). The ${\beta}$-galactosidase reporter gene showed maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Both 40 mM and 100 mM concentrations of nitrate ion in the medium had little effect on expression level. We propose that aeg-46.5 is subject to multiple regulations of anaerobic activation by Fnr, nitrate activation by NarP and repression mediated by NarL.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.218-225
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• 2007

A lactic acid bacterium capable of anaerobic respiration was isolated from soil with ferric iron-containing glucose basal medium and identified as L. garvieae by using 16S rDNA sequence homology. The isolate reduced ferric iron, nitrate, and fumarate to ferrous iron, nitrite, and succinate, respectively, under anaerobic $N_2$ atmosphere. Growth of the isolate was increased about 30-39% in glucose basal medium containing nitrate and fumarate, but not in the medium containing ferric iron. Specifically, metabolic reduction of nitrate and fumarate is thought to be controlled by the specific genes fnr, encoding FNR-like protein, and nir, regulating fumarate-nitrate reductase. Reduction activity of ferric iron by the isolate was estimated physiologically, enzymologically, and electrochemically. The results obtained led us to propose that the isolate metabolized nitrate and fumarate as an electron acceptor and has specific enzymes capable of reducing ferric iron in coupling with anaerobic respiration.

• 한국환경보건학회지
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• 제38권6호
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• pp.568-574
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• 2012

Objectives: The purpose of this experiment is to understand the phosphorus removal ratio effects of iron plates per unit of surface area through the iron electrolysis system, which consists of an anoxic basin, aerobic basin, and iron precipitation apparatus. Methods: Iron electrolysis, which uses an iron precipitation reactor in anoxic and oxic basins, consisted of iron plates with total areas of 400 $cm^2$, 300 $cm^2$ and 200 $cm^2$ respectively. The FNR process was operated with a hydraulic retention time and a sludge retention time of 12 hours and three days, respectively. Wastewater used in the experiments was prepared by dissolving $KH_2PO_4$ in influent water. Results: The iron plates 400 $cm^2$ (16.6 $mA/cm^2$), 300 $cm^2$ (13.3 $mA/cm^2$) and 200 $cm^2$ (7.3 $mA/cm^2$) in surface area in the phosphorus reactor had respective phosphorus of 2.4 mg/l, 2.7 mg/l and 3.2 mg/l in the effluent and phosphorus removal respective efficiencies of 90.3%, 89.1% and 87.1%. The effluent in the reactor, where the iron plate was not used, had relatively very low phosphorus removal efficiency showing phosphorus concentration of 15.3 mg/l and a phosphorus removal efficiency about 38.3%. Phosphorus removal per ferrous was 0.472 mgP/mgFe in the iron electrolysis system where the surface area of iron was low. Phosphorus pollution load per active surface area and the phosphorus removal efficiency had an interrelation of RE = -0.27LS + 89.0 (r = 0.85). Conclusion: With larger iron plate surface area, the elution of iron concentration and phosphorus removal efficiency was higher. The removal efficiency of phosphorus has decreased by increasing the initial phosphate concentration in the iron electrodes. This shows a tendency of decreasing phosphorus removal efficiency because of decreasing of iron deposition as the phosphorus pollution load per active surface area increases.

• 생명과학회지
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• 제16권4호
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• pp.632-639
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• 2006

본 연구에서는 lacZ transcriptional fusion plasmid를 이용하여 광합성 세균인 Rhodobacter sphaeroides에서의 7가지 광합성유전자 (puf, puc, puhA, bchC, bchE, bchF, bchI) 발현의 경향과 조절을 조사하였다. R. sphaeroides에서 puhA와 bchI를 제외한 모든 광합성유전자들이 호기적 조건과 비교했을 때 혐기적 조건에서 더욱 강하게 발현되었다. puhA 유전자는 bchFNBHLM-RSP0290과 operon을 형성하며, bchI 유전자는 crtA와 operon을 이루는 것으로 나타났다. 광합성 조건에서 자란 R. sphaeroides의 puf, puc, bchCXYZ operon의 발현은 빛의 세기에 비례하는 반면, bchFNBHLM(RSP0290 puhA) operon의 발현은 빛의 세기에 반비례 하였다. bchEJG의 발현은 $10\;W/m^2$의 빛이 조사된 광합성 조건에서 제일 낮았으며, $100\;W/m^2$의 빛의 광합성 조건에서 가장 높았다. R. sphaeroides의 산소인지와 빛 인지에 관련된 세 가지 주요 조절기작에 의한 광합성유전자 조절은 다음과 같다. puf와 bchC는 PpsR repressor와 PrrBA two-component system에 의해 조절된다. 그리고 puc operon은 PpsR, FnrL, PrrBA system에 의해 조절된다. bchE의 발현은 FnrL과 PrrBA system에 의해 조절되는 반면, bchF는 오로지 PpsR에 의해서만 조절된다. PpsR repressor는 강한 세기의 빛 조건에서 bchf 발현억제의 원인이 되며, FnrL은 그 자체가 산소를 인지하는 기능 이외에도 세포질의 산화/환원 상태의 인지에 관련될 것으로 보인다.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.436-442
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• 1999

The aeg-46.5 operon of Escherichia coli is induced by nitrate and anaerobic conditions. Positive regulators Fnr and NarP, and a negative regulator NarL control the expression of the aeg-46.5. It has two symmetry regions [6], one of which is located between +37 and +56 bp from the 5'end of the anaerobic transcription initiation site. In this study, mutagenized symmetry regions were transferred from plasmid to chromosome by homologous recombination to evaluate the mutation as a single copy in the fnr, narL, narP, and narL-narP double mutant background. The expressions of the aeg-46.5 operon with these mutations indicated that the control was not through the possible stem-loop structure. Whether there is a protein that mediates this control remains to be seen. The results from the narL-narP double mutant indicated that the anaerobic Fill induction was independent of NarL repression.

• BMB Reports
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• 제29권1호
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• pp.92-97
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• 1996

The age-46.5 gene of Escherichia coli is induced by nitrate ion and regulated by Fnr, NarL, and NarP during anaerobic growth. aeg-46.5::lacZ fusion gene shows its maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Fnr and NarP act as positive regulators, and NarL acts as a negative regulator. The control region of the aeg-46.5 was identified and the binding sites of regulator proteins have been predicted (Reznikoff and Choe (1993)). It has two symmetry regions. One is located at -52~-37 bp from the anaerobic mRNA 5'-end, which is the binding site of NarL and NarP. The other is located at +37~+56 bp from the 5'-end of mRNA. In this study, the downstream symmetry region from the mRNA 5'-end was investigated by site-directed mutagenesis. The destruction of the symmetry region increases the expression level of aeg-46.5. We propose that the symmetry region interferes with the expression of aeg-46.5 possibly by forming a stem-and-loop structure.

• 한국환경보건학회지
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• 제39권1호
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• pp.83-89
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• 2013

Objectives: The purpose of this experiment was to illuminate the relationship between the phosphorus removal rate of unit operation and the phosphorus removal rate of phosphorus volume loading in the Ferrous Nutrient Removal process, which consists of an anoxic basin, oxic basin, and iron precipitation apparatus. Methods: This study was conducted in order to improve the effect of nitrogen and phosphorus removal in domestic wastewater using the FNR (Ferrous Nutrient Removal) process which features an iron precipitation reactor in anoxic and oxic basins. The average concentration of TN and TP was analyzed in a pilot plant ($50m^3/day$). Results: The removal rate of T-N and T-P were 66.5% and 92.8%, respectively. The $NH_3-N$ concentration of effluent was 2.62 mg/l with nitrification in the oxic basin even though the influent was 17.7 mg/l. The $NO_3$-N concentration of effluent was 5.83 mg/l through nitrification in oxic basin even though the influent and anoxic basin were 0.82 mg/l and 1.00 mg/l, respectively. The specific nitrification of the oxic basin ($mg.NH_3$-Nremoved/gMLVSSd) was 16.5 and specific de-nitrification ($mg.NO_3$-Nremoved/gMLVSSd) was 90.8. The T-P removal rate was higher in the oxic basin as T-P of influent was consumed at a rate of 56.3% in the anoxic basin but at 90.3% in the oxic basin. The TP removal rate (mg.TP/g.MLSS.d) ranged from 2.01 to 4.67 (3.06) as the volume loading of T-P was increased, Conclusions: The test results showed that the electrolysis of iron is an effective method of phosphorus removal. Regardless of the temperature and organic matter content of the influent, the quality of phosphorus in the treated water was both relatively stable and high due to the high removal efficiency. Nitrogen removal efficiency was 66.5% because organic matter from the influent serves as a carbon source in the anoxic basin.

• 대한토목학회논문집
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• 제4권4호
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• pp.25-36
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• 1984

본(本) 연구(硏究)는 강구조부재(鋼構造部材)의 안정성수준(安全性水準)을 LRFD에 의해 검토(檢討)하였다. 그리고, 우리 현실(現實)을 고려(考慮)한 목표신뢰성지수(目標信賴性指數)(${\beta}_0$)를 설정(設定)하여 이에 대한 저항(抵抗) 및 하중계수(荷重係數)를 방법(方法)으로 계산(計算)하였으며, 각(各) 도로교시방서(道路橋示方書) 공칭안전율(公稱安全率)과 이들 계산(計算)된 계수(係數)를 비교분석(比較分析)하였다. 강구조부재(鋼構造部材)의 저항(抵抗) 및 하중효과(荷重効果)에 대한 불확실량(不確實量) 산정(算定)은 Galambos-Ravindra 및 SGST의 하중(荷重) 및 저항(抵抗) 불확실량(不確實量) 해석방법(解析方法)에 따랐다. 그 결과(結果)를 요약(要約)하면 다음과 같다. 1) 현(現) 강구조부재(鋼構造部材)에 대한 목표신뢰성지수(目標信賴性指數)(${\beta}_0$)는 우리의 현실(現實)을 고려(考慮)하여 ${\beta}_0=3.5$가 적절(適切)하다. 2) 1)에 대한 강도설계(强度設計) 방정식(方程式)에 공칭저항(公稱抵抗) ${\phi}^{\prime}$, 공칭하중계수(公稱荷重係數) ${\gamma}^{\prime}$는 아래와 같다. a) 양연지지판(兩緣支持板): ${\phi}^{\prime}=0.75$, ${\gamma}_D{^{\prime}}=1.04$, ${\gamma}_L{^{\prime}}=2.08$ b) 편지지판(片支持板): ${\phi}^{\prime}=0.82$, ${\gamma}_D{^{\prime}}=1.04$, ${\gamma}_L{^{\prime}}=2.08$.

• KSBB Journal
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• 제11권4호
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• pp.431-437
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• 1996

본 연구에서는 질산염(nitrate) 존재하에서 혐기 조건이 되면 최대로 발현되는 변형된 naT 프로모터 를 대장균내에서 단백질을 대량 생산하기 위한 발현 운반체로 쓰일 수 있는지를 조사하였다. 이를 위해 용존산소농도에 따라 naT 프로모터의 유도에 영향을 미치는 염색체 fnT 유전자가 발현되지 못하는 대장 균(ES200l)에 pBR322 플라스미드에 프로모터상 의 10 지역에서 특정부위돌연변이화에 의해 con­s sensus sequence로 바핀 변형된(modified) naT 프 로모터(pMW616)가 도입되어 있는 계(pMW616/ E ES2001 )가 이용되었다. 이 변형된 naT 프로모터의 하류에는 구조유전자(structural gene) 인 질산염 환 원효소대 신에 ${\beta}$-galactosidase를 발현하는 lacZ 유 전자가 클로닝되어 있다. 이 변형된 naT 프로모터의 유도에 대한 최적조건을 찾기 위해 변형된 naT 프로 모터를 유도시키는 방법, 변형된 naT 프로모터가 최 대로 유도되는 질산엽의 농도, 말현된 $\beta$galactosid a ase의 양 빛 변형된 naT 프로모터가 유도되는 특성 들이 조사되었다. 이에 대한 실험으로부터 다음과 같은 결론들이 얻어졌다; 이 변형된 naT 프로모터로 부터 ${\beta}$-galactosidase의 말현은 질산염의 농도에 의 해서는 크게 영향을 받지 않았다. 한편, 변형된 naT 프로모터로부터 ${\beta}$-galactosidase의 말현은 성장단계 에서 대장균을 호기상태에서 OD600이 약 2.27~ 될 때까지 키우다가 유도시 혐기상태로 만드는 것이 가 장 유리하였으며, 이 때 발현된 ${\beta}$-galactosidase의 비활성도는 약 13,000 Miller unit였다. 하지만, 이 변형된 naT 프로모터는 이켓을 유도시키기 전에도 발현된 ${\beta}$-galactosidase의 비활성도가 약 6,000 M Miller unit로 매우 높음으로 인하여 이 변형된 naT 프로모터는 원하는 단백질을 유도성보다는 구성적으 로 발현시키는데 더 적합한 프로모터였다.