• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.158-158
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• 1998

The non-mevalonate pathway is a newly discovered isoprenoid biosynthetic pathway in some bacteria, cyanobacteria, algae and plants. Because isoprenoid metabolites (ubiquinone, menaquinone, undecaprenol) are essential for bacterial growth, this pathway may represent a novel target for antibacterial agents. Antibiotics with a unique mechanism of action are needed to combat the risk of antibiotic resistance that is a current worldwide problem. In order to study this pathway as viable target, it was necessary to verify use of the pathway in our model system, the bacterium Bacillus subtilis. Incubation experiments with [6,6-$^2$H$_2$]-D-glucose and [l-$^2$H$_3$]-deoxy-D-xylulose were conducted to provide labeled menaquinone-7 (MK -7), the most abundant isoprenoid in B. subtilis. $^2$H-NMR analysis of the MK-7 revealed labeling patterns that strongly support utilization of the non-mevalonate pathway. Another approach to study the pathway is by structure activity relationships of proposed inhibitors of the pathway. Fosmidomycin is a phosphonic acid with antibacterial activity known to inhibit isoprenoid biosynthesis in susceptible bacteria and may act by inhibiting the non-mevalonate pathway. Fosmidomycin and an N-methyl analog were synthesized and tested for antibacterial activity. Fosmidomycin was active against Escherichia coli and B. subtilis, while N-formyl-N-methyl-3-amino-propylphosphonic acid was inactive.

• Journal of Microbiology
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• 제44권2호
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• pp.171-176
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• 2006

A strictly aerobic, non-motile, ovoid-shaped Alphaproteobacteria, designated strain $JC2049^T$ was isolated from a tidal flat sediment sample. The results of 16S rRNA gene sequence analysis indicated that this isolate belonged to the genus Thalassobius, with a sequence similarity of 96.9-97.3% to other valid Thalassobius spp. The cells required 1-7% NaCl for growth (optimum 2%) and accumulated $poly-\beta-hydroxybutyrate$. Nitrite was reduced to nitrogen, but nitrate was not reduced to nitrite. No genetic potential for aerobic anoxygenic photosynthesis was detected. The primary isoprenoid quinone (Ubiquinone-10), predominant cellular fatty acids $(C_{18:1}{\omega}7c,\;11\;methyl\;C_{18:1}\omega7c\;and\;C_{16:0})$ and DNA G+C content (61 mol %) were all consistent with the assignment of this isolate to the genus Thalassobius. Several phenotypic characteristics clearly distinguished our isolate from other Thalassobius species. The degree of genomic relatedness between strain $JC2049^T$ and other Thalassobius species was in a range of 20-43 %. The polyphasic data presented in this study indicates that our isolate should be classified as a novel species within the genus Thalassobius. The name Thalassobius aestuarii sp. novo is therefore proposed for this isolate; the type strain is $JC2049^T(=IMSNU\;14011^T=KCTC\;12049^T=DSM\;15283^T)$.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.643-648
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• 2002

For wastewater treatment and utilization of the biomass, a photosynthetic bacterium was isolated based on its cell growth rate, cell mass, and assimilating ability of organic acids. The isolate was a Gram-negative rod-shaped bacterium that contained a single polar flagellum and formed a lamellar intracytoplasmic membrane (ICM) system, including bacteriochlorophyll $\alpha$. The major isoprenoid quinone component was identified as ubiquinone Q-10, and the fatty acid composition was characterized as to contain relatively large amount of C-16:0 (18.74%) and C-18:1 (59.23%). Based on its morphology, phototrophic properties, quinone component, and fatty acid composition, the isolate appeared to be closely related to the Rhodopseudomonas subgroup of purple nonsulfur bacteria. A phylogenetic analysis of the isolate using its 16S rRNA gene sequence data also supported the phenotypic findings, and classified the isolate closely related to Rhodopseudomonas palustris. Accordingly, the nomenclature of the isolate was proposed as Rhodopseudomonas palustris KUGB306. A bench-scale photosynthetic bacteria (PSB) reactor using the isolate was designed and operated for the treatment of soybean curd wastewater.

• 농업과학연구
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• 제19권1호
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• pp.28-32
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• 1992

태국의 토양으로부터 분리한 젤라틴 분해능을 갖는 광합성세균 T-20주(株)는 직경이 $0.6{\sim}0.7{\mu}m$의 적색 간균으로써, Bacteriochlorophyll a의 흡수극대는 870nm이었고 또한, 퀴논종(種)은 UQ-8을 나타내었다. 유기 화합물로써 citrate, aspirate, glutamate, fractose 등을 이용하였다. 이상의 결과로부터 T-20주는 Bergey's Manual of Systematic Bacteriology(1989)에 기재된 Rhodocyclus gelatinosus와 일치한다.

• 미생물학회지
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• 제29권4호
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• pp.250-257
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• 1991

An obligate methanol-oxidizing bacterium, Methylobacillus sp. strain SK1, which grows only on methanol was isolated from soil. The isolate was nonmotile Gram-negtive rod. It does not have internal membrane system. The colonies were small, whitish-yellow, and smooth. The guanine plus cytosine content of the DNA was 48 mol%. Cellular fatty acids consisted predominantly of large amounts of straight-chain saturated $C_{16:0}$ acid and unsaturated $C_{16:1}$ acid. The major ubiquinone was Q-8, and Q-10 was present as minor component. The cell was obligately aerobic and exhibited catalase, but no oxidase, activity. Poly-.betha.-hydroxybutyrate, endospores, or cysts were not observed. the isolate could grow only on methanol in mineral medium. Growth factors were not required. The isolate was unable to use methane, formaldehyde, formate, methylamine, and several other organic compounds tested as a sole source of carbon and energy. Growth was optimal at 35.deg.C and pH 7.5. It could not grow at 42.deg.C. The doubling time was 1.2h at 30.deg.C when grown with 1.0%(v/v) methanol. The growth was not affected by antibiotics inhibiting cell wall synthesis and carbon monoxide but was completely suppressed by those inhibiting protein synthesis. Methanol was found to be assimilated through the ribulose monophosphate pathway. Cytochromes of b-, c-, and o- types were found. Cell-free extracts contained a phenazine methosulfate-linked methanol dehydrogenase activity, which required ammonium ions as an activator. Cells harvested after the late exponential phase seemed to contain blue protein.ein.

• 상하수도학회지
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• 제16권6호
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• pp.679-685
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• 2002

The objective of this study was investigated for the microbial community structure and treatment performance of domestic wastewater in lab-scale submerged membrane bioreactor operated with anoxic-oxic cycles. Respiratory quinone profiles were applied as tools for identifying different bacterial populations. The cycle time program of bioreactor was control under anoxic/oxic of 60/90 minutes with an hydraulic retention time of 8.4 hrs. The average $COD_{Cr}$ removal efficiency of domestic wastewater was as high as 93%. The results showed complete nitrification of $NH_4^+$-N generated during oxic period and up to 50% of the total nitrogen could be denitrified. The dominant quinone types of suspended microorganisms in bioreactor were ubiquinone (UQ)-8, -10, followed by menaquinone (MK)-6, and MK-7 for anoxic period, but those for oxic period were UQ-8, MK-6, followed by UQ-10 and MK-7. The microbial diversities of bioreactor at anoxic and oxic periods, calculated based on the composition of all quinones were 10.4 and 12.2-11.8, respectively. The experimental results showed that the microbial community structure in the submerged membrane bioreactor treating domestic wastewater was slightly affected by intermittent aeration.

• 한국수정란이식학회지
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• 제25권1호
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• pp.35-42
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• 2010

Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.

• Journal of Microbiology and Biotechnology
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• 제13권1호
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• pp.104-109
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• 2003

Enhancement of the production of benzoylformate reductase (BFR) was attempted under oxidative stress established by natural electron carriers. -lipoic acid (LA), flavin adenine dinucleotide (FAD), and ubiquinone (UQ) did not inhibit growth of E. faecalis when their concentrations were as high as $10{\mu}M$, while $H_2O_2$ and methyl viologen ($MV^2+$) inhibited the bacterial growth. BFR activity in the bacterial extract had increased rapidly after 1 h of cultivation after the addition of $4{\mu}M$ of natural electron carriers, and the activity was maintained during further cultivation. BFR activity of the cells treated with the natural electron carriers was $40\%$ higher than that of the control. In the presence of $4{\mu}M\;H_2O_2\;and\;MV^2+$, BFR activity increased, reaching the highest activity at about 5 h cultivation, and then decreased with further cultivation. It seems that natural electron carriers not only stimulate the induction of BFR, but also stabilize the enzyme. BFR was hardly affected by LA, FAD, and UQ, while $H_2O_2\;and\;MV^2+$ inactivated the crude enzyme. The decrease of BFR activity in the presence of $H_2O_2\;and\;MV^2+$ might be ascribed to inactivation of the enzyme by the oxidants.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1679-1684
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• 2014

Acetobacter sp. strains were isolated from traditional vinegar collected in Daegu city and Gyeongbuk province. The strain KJY8 showing a high acetic acid productivity was isolated and characterized by phenotypic, chemotaxonomic, and phylogenetic inference based on 16S rRNA sequence analysis. The chemotaxonomic and phylogenetic analyses revealed the isolate to be a strain of Acetobacter pomorum. The isolate showed a G+C content of 60.8 mol%. It contained $\small{LL}$-diaminopimelic acid ($\small{LL}$-$A_2pm$) as the cell wall amino acid and ubiquinone $Q_9$ (H6) as the major quinone. The predominant cellular fatty acids were $C_{18:1}w9c$, w12t, and w7c. Strain KJY8 grew rapidly on glucose-yeast extract (GYC) agar and formed pale white colonies with smooth to rough surfaces. The optimum cultivation conditions for acetic acid production by the KJY8 strain were $20^{\circ}C$ and pH 3.0, with an initial ethanol concentration of 9% (w/v) to produce an acetic acid concentration of 8% (w/v).

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1743-1750
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• 2007

Two Gram-negative, nonmotile, coccobacilli, SW-$3^T$ and SW-$100^T$, were isolated from sea water of the Yellow Sea in Korea. Strains SW-$3^T$ and SW-$100^T$ contained ubiquinone-9 (Q-9) as the predominant respiratory lipoquinone and $C_{18:1}\;{\omega}9c$ and $C_{16:0}$ as the major fatty acids. The DNA G+C contents of strains SW-$3^T$ and SW- $100^T$ were 44.1 mol% and 41.9 mol%, respectively. A neighbor-joining tree based on l6S rRNA gene sequences showed that the two isolates fell within the evolutionary radiation enclosed by the genus Acinetobacter. Strains SW-$3^T$ and SW-$100^T$ exhibited a l6S rRNA gene similarity value of 95.7% and a mean DNA-DNA relatedness level of 9.2%. Strain SW-$3^T$ exhibited l6S rRNA gene sequence similarity levels of 93.5-96.9% to the validly described Acinetobacter species and fifteen Acinetobacter genomic species. Strain SW-$100^T$ exhibited l6S rRNA gene sequence similarity levels of less than 97.0% to the other Acinetobacter species except Acinetobacter towneri DSM $14962^T$ (98.0% similarity). Strains SW-$3^T$ and SW-$100^T$ exhibited mean levels of DNA-DNA relatedness of 7.3-l6.7% to the type strains of some phylogenetically related Acinetobacter species. On the basis of phenotypic, phylogenetic, and genetic data, strains SW-$3^T$ and SW-$100^T$ were classified in the genus Acinetobacter as two distinct novel species, for which the names Acinetobacter marinus sp. novo (type strain SW-$3^T$=KCTC $12259^T$=DSM $16312^T$) and Acinetobacter seohaensis sp. novo (type strain SW-$100^T$=KCTC $12260^T$=DSM $16313^T$) are proposed, respectively.