• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1701-1707
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• 2016

Lipoxygenase (LOX) is an industrial enzyme with wide applications in food and pharmaceutical industries. The available structure information indicates that eukaryotic LOXs consist of N terminus β-barrel and C terminus catalytic domains. However, the latest crystal structure of Pseudomonas aeruginosa LOX shows it is significantly different from those of eukaryotic LOXs, including the N-terminal helix domain. In this paper, the functions of this N-terminal helix domain in the soluble expression and catalysis of P. aeruginosa LOX were analyzed. Genetic truncation of this helix domain resulted in an insoluble P. aeruginosa LOX mutant. The active C-terminal domain was obtained by dispase digestion of the P. aeruginosa LOX derivative containing the genetically introduced dispase recognition sites. This functional C-terminal domain showed raised substrate affinity but reduced catalytic activity and thermostability. Crystal structure analyses demonstrate that the broken polar contacts connecting the two domains and the exposed hydrophobic substrate binding pocket may contribute to the insoluble expression of the C terminus domain and the changes in the enzyme properties. Our data suggest that the N terminus domain of P. aeruginosa LOX is required for its soluble expression in E. coli, which is different from that of the eukaryotic LOXs. Besides this, this N-terminal domain is not necessary for catalysis but shows positive effects on the enzyme properties. The results presented here provide new and valuable information on the functions of the N terminus helix domain of P. aeruginosa LOX and further improvement of its enzyme properties by molecular modification.

• 청정기술
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• 제29권2호
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• pp.135-144
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• 2023

Pseudomonas aeruginosa와 Staphylococcus aureus는 만성 창상 감염의 원인이 되는 주요 병원체이며 상처부위에서 공존한다. 이러한 감염은 단일 감염에 비해 독성이 높아 환자에게 바람직하지 않은 결과를 초래한다. 복합미생물 감염내에서 미생물 간의 상호작용은 질병 진행을 악화시키는 것으로 알려져 있다. 호흡기, 상처, 당뇨병 발과 같은 질병 내 복합 미생물 감염은 다양한 미생물들을 포함하나, 녹농균과 황색포도상구균이 가장 일반적으로 확인된다. 본 연구는 그람음성균 P. aeruginosa와 그람양성균 S. aureus를 중심으로 독성화학물질의 농도구배에 따른 성장양상을 비교하고자 하였다. 세균 성장이 억제되는 농도를 의미하는 최소 억제 농도(MIC)는 특정화학물을 함유한 배지를 연속적으로 희석하여 성장 곡선을 평가하여 결정한다. 두 균주 모두 상기 방법을 통해 성장곡선을 확인하였고, 지수성장기를 적용하여 세균의 배가시간을 계산하였다. 각 독성 물질에 대한 MIC 결과로부터 그람양성균과 그람음성균 사이의 성장 속도 차이와 각 독성 물질에 대한 내성 차이를 식별하였다. 우리는 이 접근 방식이 세균 관련 감염의 혁신적인 치료법 개발에 대한 강력한 잠재력을 가지고 있다고 기대한다.

• Archives of Pharmacal Research
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• 제12권2호
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• pp.114-118
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• 1989

The effects of intraperitoneal injection of cephradine in a dose of 75 mg/kg and netilmicin in dose of 50 mg/kg and their combination on creatinine and urea serum levels of rabbits were studied as well as the antipseudomonal activity against three multiresistant clinicial isolates. The antibacterial activity was investigated by two methods: Checkerboard titration method and time-kill studies. Finally, the antibacterial activity of the sera obtained from the rabbits receiving the used drugs in the previous regimen was studied using time-kill study method against Pseudomonas aeruginosa isolates. Results obtained from this study indicated that both creatinine and urea serum levels of the rabbits receiving both drugs were not significantly different from those of the rabbits receiving either cephradine or netilmicin alone. At the same time the in vitro antibactrial activity (either of the prepared solutions of the used drugs and their combination or of the sera obtained from the rabbits receiving these drugs as mentioned before) showed a synergistic effect against the tested strains of Pseudomonas aeruginosa

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.378-381
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• 2000

The optimal conditions and properties for the immobilization of marine bacterium Pseudomonas aeruginosa BYK-2 have been determined. For the high productioon of biosurfactant, Na-alginate, PVA, modified PVA were used as a carrier. The optimal emulsifying activity on immobilized Pseudomonas aeruginosa BYK-2 showed 1036Unit (about 2.2g/L biosurfactant) in Basal salt medium(B.S.M.) at $25^{\circ}C$, 100rpm. Ca-alginate was selected the optimal bead among PVA, modified PVA and Ca-alginate. The optimal cell load in alginate bead was 10 gCWW/100g carrier. As the results of incubation of immobilized 5g Ca-alginate bead (conditions; 3% alginate, bead diameter: 2.3mm, 10% cell load) in 50m1 production medium, The emulsifying activity of 1407Unit, about 3.0g/L biosurfactant was obtained from immobilized cell after cultivation of 92hr at $25^{\circ}C$, 100rpm.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.645-649
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• 1998

A glycolipid type of biosurfactants was obtained from a strain which had been isolated from soil. The cell was identified as Pseudomonas aeruginosa from taxonomic characteristics and was designated as YPJ80. Thin layer chromatography and deoxyhexose detection tests were done to verify the type of biosurfactant. Critical micelle concentration (CMC) of the surfactant was observed to be 50 ppm and the minimum surface tension was 30.1 mN/m. As an emulsifier, YPJ80 biosurfactant was superior to emulsan in the emulsification of crude Arabian light oil.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.355-361
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• 2001

A Chromate tolerant strain of Pseudomonas aeruginosa isolated from the effluent of a tannery showed significant enzymatic activity of chromate reduction. Cells grown in chromate-supplemented medium reduced 8 $\mu\textrm{g}$ chromate/mg protein/h in the presence of NADH/NADPH. The chromate reducing activity was inducible as cells pregrown in chromate showed higher chromate reduction. In contrast, the periplasmic fraction of cells gown in chromate reduce $75\%$ chromate in 4 h and the spheroplast fraction failed to do so, indicating that chromate reductase may be located in the periplasm. The presence of a 30 kDa protein in the periplasmic extracts of cells grown in the presence of chromate, but its absence of the protein in cells grown without chromate, points out a possible role of this protein in chromate reduction.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.347-352
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• 1998

A new strain of Pseudomonas aeruginosa growing in a rice field contaminated with herbicide and effluents of a factory manufacturing explosives was isolated. This isolate showed excellent growth in unusually high concentration of acrylamide (60 mM). It utilized acrylamide as the sole source of carbon and nitrogen for growth. Other amides such as acetamide, butyramide, isobutyramide, and methacrylamide were also utilized for the growth by this isolate. Acrylamide was degraded into acrylic acid and ammonia by the enzyme amidase. More than $65\%$ of added acrylamide (40 mM) was converted into acrylic acid after 40 h of growth of the culture. Amidase activity was inducible, the highest activity being observed with isobutyramide ($12.5{\mu}M$ ammonia/mg protein/min). These results demonstrate that this bacterium can degrade a variety of amides.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2001년도 추계학술발표회
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• pp.235-238
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• 2001

Monod kinetics에 관련된 주요 생분해 파라미터를 도출하기 위하여 microcosm 규모의 배치실험에서 BTEX 화합물에 대해 분해능이 우수한 Pseudomonas aeruginosa을 이용해 다양한 농도의 벤젠에 대한 분해기작을 고찰하였다. 벤젠의 생분해율(D)과 Maximumspecific growth rate ($\mu$$_{max}$)는 기질의 농도가 증가할수록 높아지다가 최고점에 도달 후에 점차적으로 감소하였으며 이것은 어느 한계점 이상의 벤젠 농도가 미생물의 생분해에 방해 요소로 작용한다는 것을 나타낸다. 그러나 미생물에 의한 벤젠 분해의 상관관계를 나타내는 yield coefficient(Y)는 벤젠의 초기 농도가 낮을수록 높은 값을 나타내었다. Microbial decay constant( b)와 half-saturation constant(K$_{c}$)는 각각 0.21~0.48day$^{-1}$와 218mg/$\ell$로서 문헌값 보다 높은 수치를 나타내었다. 실험으로부터 결정된 생분해 파라미터들은 초기 벤젠 농도에 따라 큰 차이를 보이므로 생분해 모델링에 사용할 파라미터는 기질농도에 따라 적절하게 선택되어야 한다고 사료된다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.626-629
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• 2000

The Pseudomonas aeruginosa BYK-2(KCTC 18012p) produced three kinds of glycolipids on olive oil as a substrate and purified two types of major glycolipids(Rf=0.48, BS-1; Rf=0.65, BS-2) using silica gel chromatography, TLC, HPLC, etc. From the analysis of the chemical structure, the glycolipid of BS-1 was estimated as rhamnolipid($2-O-{\alpha}-L-rhamnopyranosyl- {\alpha}-L-rhamnopyranosyl-{\beta}-hydroxyldecanoyl-{\beta}-hydroxydecanoic$ acid; M.W. 650) and BS-2 was detected as rhamnolipid methyl ester($2-O-{\alpha}-L-rhamnopyranosyl-{\alpha}-L-rhamnopyranosyl-{\beta}-hydroxyldecanoyl-{\beta}-hydroxydecanoic$ acid methyl ester; M.W. 664) by FT-IR, FAB Mass spectrometry, $^1H-NMR$, $^{13}C$ FT-NMR, DEPT, 2D-NMR (TOCSY, RELAY, NOESY, HSQC, HMBC). In particular, It was found that a marine bacterium Pseudomonas aeruginosa BYK-2(KCTC 18012P) remarkably produced rhamnolipid and rhamnolipid methyl ester simultaneously.

• 센서학회지
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• 제21권5호
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• pp.374-378
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• 2012

Biofouling causes many problem in industrial processes, medical health industries, water utilities and our daily life. So detecting formation of biofilm on the surface of medical appliance, water pipe and industrial utility is highly important to prevent the problem caused by biofouling. In this study, we suggest an electrochemical sensor for detecting biofilm. We fabricated the electrochemical sensor in MEMS process and cultivated two different kinds of Pseudomonas aeruginosa RpoN type and Wild type on the surface of electrochemical sensor. Each group of Pseudomonas aeruginosa was cultivated according to the hours of 2, 4, 6, 8, 12 and 24. Then we investigated changes in degree of biofilm cultivation using cyclic voltammetry. As a result, it was observed that peak of the cyclic voltammetry curve is increased according as the biofilm growth on the surface of electrochemical sensor. Also we can discern between Pseudomonas aeruginosa RpoN type and Wild type.