• 대한의생명과학회지
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• 제17권1호
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• pp.7-12
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• 2011

Free-living amoebae ingest several kinds of bacteria. In other words, the bacteria can survive within free-living amoeba. To determine how Escherichia coli K1 isolate causing neonatal encephalitis and non-pathogenic K12 interact with free-living amoebae, e.g., Acanthamoeba castellanii (T1), A. astronyxis (T7), Naegleria fowleri, association, invasion and survival assays were performed. To understand pathogenicity of free-living amoebae, in vitro cytotoxicity assay were performed using murine macrophages. T1 destroyed macrophages about 64% but T7 did very few target cells. On the other hand, N. fowleri which needed other growth conditions rather than Acanthamoeba destroyed more than T1 as shown by lactate dehydrogenase (LDH) release assay. In association assays for E. coli binding to amoebae, the T7 exhibited significantly higher association with E. coli, compared with the T1 isolates (P<0.01). Interestingly, N. fowleri exhibited similar percentages of association as T1. Once E. coli bacteria attach or associate with free-living amoeba, they can penetrate into the amoebae. In invasion assays, the K1 (0.67%) within T1 was observed compared with K12 (0%). E. coli K1 and K12 exhibited high association with N. fowleri and bacterial CFU. To determine the fate of E. coli in long-term survival within free-living amoebae, intracellular survival assays were performed by incubating E. coli with free-living amoebae in PBS for 24 h. Intracellular E. coli K1 within T1 (2.5%) and T7 (1.8%) were recovered and grown, while K12 were not found. N. fowleri was not invaded and here it was not recovered.

• 미생물학회지
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• 제38권3호
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• pp.192-197
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• 2002

러시아 바이칼호에서 해빙기에 부유세균과 aggregates에 부착한 세균의 군집구조를 FISH (fluorescent in situ hybridization)방법으로 0 m부터 250 m수심에서 비교 분석하였다. 조사대상은 Eubacteria에 속하는 세균과 class Proteobacteria에 속하는 세균 중 $\alpha$-, $\beta$-, $\gamma$ -group과 Cytophaga-Flavobacterium group,그리고 Planctomycetales였다. 부유세균의 수는 $0.2{\times}10^6\cells{\cdot}ml^-1 to 3.2{\times}10^6 \cells{\cdot}ml^-1$범위였으며, 수심이 깊어질수록 감소하였다. Aggregate에 부착한 세균은 부유세균과 반대로 수심이 깊어질수록 증가하였고, 개체수의 범위는 0.4~$3.3{\times}10^4$ $cells{\cdot}ml^-1$ 이였다. 총세균수에 대한 Eubacteria 수의 비율은 부유세균의 경우 52.3~74.1%, aggregates에 부착한 세균은 39.6~66.7%로 부유세균보다 부착세균에서 그 비율이 낮았으며, 세균의 군집구조 분석 결과에서도 부유세균과 aggregates애 부착한 세균의 군집구조가 다른 양상으로 나타났다. 특히 두 세균의 군집구조는 식물플랑크톤이 밀집해 있는 25 m 수심에서 급격히 변화하여 식물플랑크톤이 부유세균과 부착세균의 군집의 변화에 밀접한 관련이 있는 것으로 확인되었다, Aggregates에 부착한 세균 군집은 수심에 따라 매우 특이한 변화 양상을 나타내었다. $\beta$-proteobacteria group은 수심이 깊어지면서 그 비율이 높아져, 100m에서는 $\beta$-group이 총세균수는 51.8%를 차지하였으나, 250m에서는 $\gamma$-group이 43.8%를 차지하여, 급격하게 우점 group이 변화하였다. 그러나, 부유세균에서는 전혀 다른 군집 구조를 이루고 있었다. 이러한 결과에서 aggregates에 부착한 세균은 부유세균과는 다른 다양성을 이루고, 다른 천이과정을 거치는 것으로 확인되었다.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.18-24
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• 2003

벼과식물과 관련된 질소고정균을 분리하기 위하여 한반도 남부지방의 19지역에서 쌀, 밀, 귀리, 보리, 호밀, 옥수수 등을 채집하여 이들의 뿌리로부터 free living bacteria와 endophyte를 분류하였다. Colony의 형태, 크기, 색깔 등과 16S rDNA 염기서열을 근거로 63종으로 분류하였다. nifH 유전자 염기서열 결정과 nitrogenase 활성 측정을 통하여 분리균주 모두 질소고정 능력이 있음을 확인하였다. 그리고 식물에서 분리한 내생균의 특징 중 하나인 cellulase활성이 대부분의 분리 균주에서 확인되었다.

• Ocean and Polar Research
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• 제39권4호
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• pp.245-255
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• 2017

There are a number of pieces of evidences that suggest a link between marine diatoms and microorganisms, but knowledge about related microbial communities is greatly lacking. The present study investigated the microbial community structures related to the growth of the marine diatom Cyclotella meneghiniana. We collected free-living bacteria (FLB) and particle-associated bacteria (PAB) at each growth stage (e.g., lag, exponential, stationary and death) of the diatom, and analyzed their bacterial 16S rDNA using pyrosequencing. Metagenomics analysis showed that community structures of FLB and PAB differed considerably with the progress of growth stages. FLB showed higher diversity than PAB, but variation in the different growth stages of C. meneghiniana was more evident in PAB. The proportion of the genus Hoeflea, belonging to the order Rhizobiales, was dominant in both FLB and PAB, and it gradually increased with the growth of C. meneghiniana. However, Enhydrobacter clade tended to considerably decrease in PAB. In addition, Marinobacter decreased steadily in FLB, but first increased and then decreased in PAB. These results suggest that Hoeflea, Enhydrobacter, and Marinobacter may be closely related to the growth of diatom C. meneghiniana.

• Journal of Microbiology
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• 제42권4호
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• pp.357-360
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• 2004

For determining the process of bacterial aggregation, glucose was added into water from Lake Baikal which had been stored for seven months. In the presence of a higher concentration of glucose, the abun­dance of single bacteria and aggregates were higher, but the biovolumes of both bacteria were similar. Theses results mean that both free-living and aggregated bacteria have similar maximum sizes and that aggregates are forming with available organic materials. With available organic materials, the biovol­ume of aggregates becomes larger.

• Journal of Applied Biological Chemistry
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• 제42권1호
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• pp.1-6
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• 1999

Poly-${\beta}$-hydroxybutyrate (PHB) and enzymes related PHB metabolism have been measured in nitrogen-fixing symbiosis of chickpea and cowpea plants. Bacteroids from chickpea and cowpea contained PHB to 0.8% and 43% of their dry weight, respectively, whereas the free-living cells CC 1192 and I 16 produced $285{\pm}55mg$ and $157{\pm}18mg$ of PHB g (dry weight)$^{-1}$. To further understand why chickpea bacteroids contained little PHB, the enzyme activities of PHB metabolism (3-ketothiolase, acetoacetyl-CoA reductase, PHB depolymerase, and 3-hydroxybutyrate dehydrogenase), the TCA cycle (malate dehydrogenase, citrate synthase, and isocitrate dehydrogenase), and related reactions (malic enzyme, pyruvate dehydrogenase, and glutamate:2-oxoglutarate transaminase) were compared in extracts from chickpea and cowpea bacteroids and the respective free-living bacteria. Significant differences were observed between chickpea and cowpea bacteroids and between the bacteroid and free-living forms of CC 1192, with respect to the capacity for some of these reactions. It is indicated that a greater potential for oxidizing malate to oxaloacetate in chickpea bacteroids could be a factor that favors the utilization of acetyl-CoA in TCA cycle rather than for PHB synthesis.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.303-309
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• 1985

Continuous production of hydrogen by Ca alginate-immobilized photosynthetic bacteria was studied in a packed-bed bioreactor. The dilution rate and input concentration of carbonaces substrate were selected as operating parameters. To choose the strain for immobilization, hydrogen productivities of Rhodopseudomonas caposulata 10006 and Rhodospirillum rubrum KS-301 were compared through preliminary batch cultures of their free cells: the former was found to show better hydrogen productivity in spite of its lower specific growth rate. For the continuous production of hydrogen by immobilized R capsulata, the optimum dilution rate was about 0.84 h$^{-1}$ . The Immobilized tells gave better hydrogen yield and conversion efficiency than free ones. And a kinetic parameter K'$_{m}$ was determined for the packed-bed bioreactor, being practically constant for a specific range of dilution rates.s.

• 미생물학회지
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• 제21권2호
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• pp.71-78
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• 1983

The number of aerobic free-living nitrogen fixation bacteria and factors in soil at different stands covered with Pinus rigida, Quercus acutissima and Zoysia japonica in Cheongju area were investigated from Feb. to Sept 1981. 1. The numbers of $N_2-fixation$ bacteria, according to the seasonal changes, increased gradually from winter to spring and summer. But the growth pattern revealed some differences in accordance with plant cover stands : the numbers increased abruptly in May at Pinus, May-June at Quercus and Apr. May at Zoysia stand. The pick of numbers represented in Aug. Sept, at Pinus, Jul-Aug. at Quercus and May-Jun. at Zoysia stand, respectively. 2. The interrelationship between the monthly changes of enviotnmental factors and numbers of $N_2-fixation$ bacteria at different stands, mainly depends upon the soil temperature than other soil factors (r=0.71-0.84). The numbers of $N_2-fixation$ bacteria may increase 5-7 times according to increase $10^{\circ}C$ of soil temperature, and optimal range was $20{\sim}30^{\circ}C$ for growth. Equation of the interrelation between soil temperature and numbers could be stated as follows : log y=ax+b. 3. In the case of high soil temperature, the bacterial numbers presented high level in drought periods. Therefore, the $N_2-fixation$ bacterial species in these soil seem to consist of resistant to desication. 4. The influence of soil organic matter for growth of $N_2-fixation$ bacteria indicated low conrelation. The reason may seen the content of organic matter in these soil existed abundantly above the quantities of limitation for growth. 5. In artifical gradients, the $N_2-fixation$ bacteria were predominated at $20{\sim}30^{\circ}C$ same as natural condition, pH7-8, and 20-30% of soil water contents. 6. The vertical distribution of bacteria marked decreasing trends from surface to lower layers, and the decreasing degree was shown well in Zoysia, Quercus and Pinus stand in order. But in the trees, the numbers increased at 30cm layer estimated the region of root than 20cm layer. 7. Both catalase megative and positive group of $N_2-fixation$ bacteria in soil increased according to the rise of the soil temperature. Catalase positive group was revealed as dominant group in winter, and catalase negative group revealed in summer. The change of dominant pattern was shown during Feb. to Apr.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.94-99
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• 2002

The oceans cover about $71\%$ of the Earth's crust and contain nearly 300,000 described species. Free-living bacteria in the sea and symbiotic bacteria of marine invertebrates are proving to be valuable sources of useful bioactive compounds. Marine sponges, in particular, which contain diverse communities of bacteria, produce many classes of compounds that are unique to the marine environment. Uncultured microorganisms are commonly believed to represent $99.9\%$ of the whole microbial community. They have been investigated for the possibility of isolating and over-expressing genes in viable microorganisms. Strict symbiotic species that have been adapted to the host are candidate unculturable species. With the enormous potential for discovery, development, and market value of marine derived compounds, supply of the products is a major limiting factor for further development.

• BMB Reports
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• 제37권1호
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• pp.83-92
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• 2004

As a result of the enormous amount of information that has been collected with E. coli over the past half century (e.g. genome sequence, mutant phenotypes, metabolic and regulatory networks, etc.), we now have detailed knowledge about gene regulation, protein activity, several hundred enzyme reactions, metabolic pathways, macromolecular machines, and regulatory interactions for this model organism. However, understanding how all these processes interact to form a living cell will require further characterization, quantification, data integration, and mathematical modeling, systems biology. No organism can rival E. coli with respect to the amount of available basic information and experimental tractability for the technologies needed for this undertaking. A focused, systematic effort to understand the E. coli cell will accelerate the development of new post-genomic technologies, including both experimental and computational tools. It will also lead to new technologies that will be applicable to other organisms, from microbes to plants, animals, and humans. E. coli is not only the best studied free-living model organism, but is also an extensively used microbe for industrial applications, especially for the production of small molecules of interest. It is an excellent representative of Gram-negative commensal bacteria. E. coli may represent a perfect model organism for systems biology that is aimed at elucidating both its free-living and commensal life-styles, which should open the door to whole-cell modeling and simulation.