• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.218-223
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• 2011

To obtain eicosapentaenoic acid (EPA)-producing bacteria, some 600 strains of bacteria were isolated from Antarctic sediment and marine organisms during the summer expedition of 1999-2000 and 7 EPA-producing bacteria were obtained through screening with TLC and GC. A strain designated as L93 showed the highest EPA production, which was gram-negative, rod-shaped bacterium. L93 strain was identified as Shewanella sp., from the sequence analysis of 16S rDNA. Optimal conditions temperature and pH for the growth and EPA production were about $4^{\circ}C$ and pH 7. In addition, its production was optimized by 50%(w/v) sea salt. We establish the optimal production system to produce about 320 mg per liter by using this optimal EPA production conditions. EPA-methyl ester was purified from cultured L93 strain to a purity of higher than 97% and typical purification yield is greater than 72% of the input amount via urea complexation and HPLC.

• Economic and Environmental Geology
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• v.42 no.5
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• pp.471-477
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• 2009

An experimental removal of dissolved uranium (U) exsiting as uranyl ion (${UO_2}^{2+}$) was carried out using Shewanella p., iron-reducing bacterium. By the microbial reductive reaction, initial U concentration ($50{\mu}M$) was constantly decreased, and most U were removed from solution after 2 weeks. Major mechanism that U was removed from the solution was adsorption, precipitation and mineralization on the microbe surface. Under the transmission electron microscopy, the U adsorbed on the microbe was observed as being crystallized and eventually enlarged to several ${\mu}m$ sizes of minerals by combining with individual microbes and organic exudates. It seems that such U growth and mineralization on the microbial surface could affect the U behavior in a radioactive waste disposal site. Thus, the biogechemical reaction of metal-reducing bacteria observed in this experiment could give an affirmative measure that the microbial activity may retard U movement in subsurface environment.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.881-887
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• 2009

Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.372-378
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• 2013

An alginate-degrading bacterium, identified as Shewanella oneidensis PKA 1008 by 16S ribosomal RNA sequence analysis, was isolated from the green alga Ulva pertusa. Optimal conditions for the alginate-degrading ability of its crude enzyme were then determined. The optimal culture conditions for the growth of S. oneidensis PKA 1008 were pH 9, 2% NaCl, $30^{\circ}C$, and 24 hours incubation time. The crude enzyme produced by S. oneidensis PKA 1008 showed the highest alginate-degrading activity at pH 9, $30^{\circ}C$ and produced 1.001 g of reducing sugar per liter in 3.5% (w/v) sodium alginate for 1 hour.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.811-817
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• 2013

The present study dealt with the isolation, identification and enzyme characterization of potential luminous bacteria from water, sediment, squid, and cuttle fish samples of the Karaikal coast, Bay of Bengal, India during the study period September 2007 - August 2008. Bioluminescent strains were screened in SWC agar and identified using biochemical tests. As Shewanella henadai was found to be the most common and abundant species with maximum light emission [69,702,240 photons per second (pps)], the optimum ranges of various physicochemical parameters that enhance the luciferase activity in Shewanella hanedai were worked out. The maximum luciferase activity was observed at the temperature of $25^{\circ}C$ (69,674,387 pps), pH of 8.0 (70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps) as nitrogen source, 0.9% glycerol (71,625,196 pps), and the ionic supplements of 0.3% $CaCO_3$ (73,991,591 pps), 0.3% $K_2HPO_4$ (73,919,915 pps), and 0.2% $MgSO_4$ (72,161,155 pps). Shewanella hanedai was cultured at optimum ranges for luciferase enzyme characterization. From the centrifuged supernatant, the proteins were precipitated with 60% ammonium sulfate, dialyzed, and purified using anion-exchange chromatography, and then luciferase was eluted with 500 mM phosphate of pH 7.0. The purified luciferase enzyme was subjected to SDS-PAGE and the molecular mass was determined as 78 kDa.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.79-84
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• 2020

This study investigated the characterization and functionality of Undaria pinnatifida root (UPT) extracts, degraded using a crude enzyme from Shewanella oneidensis PKA1008. To obtain the optimum degrading conditions, the UPT was mixed with alginate degrading enzymes from S. oneidensis PKA 1008 and was incubated at 30℃ for 0, 3, 6, 12, 24, and 48 h. The alginate degrading ability of these enzymes was then evaluated by measuring the reducing sugar, viscosity, pH and chromaticity. Enzymatic extract at 24 h revealed the highest alginate degrading ability and the lowest pH value. As the incubation time increased, the lightness (L ^⁎) also decreased and was measured at its lowest value, 39.84, at 12 hours. The redness and yellowness increased gradually to 10.27 at 6 h and to 63.95 at 3 h, respectively. Moreover, the alginate oligosaccharides exhibited significant anti-inflammatory activity. These results indicate that a crude enzyme from S. oneidensis PKA 1008 can be used to enhance the polysaccharide degradation of UPT and the alginate oligosaccharides may also enhance the anti-inflammatory effect.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.510-515
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• 2006

The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.6
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• pp.888-897
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• 2015

The anti-inflammatory effect of alginate oligosaccharides on LPS-induced RAW 264.7 cells was investigated at different time points (0-60 h). The alginate oligosaccharides were produced by an alginate-degrading enzyme from Shewanella oneidensis PKA1008. The alginate oligosaccharides decreased the production of nitric oxide and proinflammatory cytokines [tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6] in a dose-dependent manner. The alginate oligosaccharides showed peak anti-inflammatory activity after 36 h of incubation; at that time point, reduced protein expression of NF-${\kappa}B$ p65, iNOS, and COX-2 was detected. Furthermore, the alginate oligosaccharide treatment reduced the formation of ear edema at 36 h compared to samples examined at 0 h when the oligosaccharides were administered at 50 and 250 mg/kg body weight, as well as dermal thickness and mast cell numbers in a histological analysis. These results suggest that alginate oligosaccharides are a promising anti-inflammatory agent.

• KSBB Journal
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• v.14 no.6
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• pp.731-736
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• 1999

87 microbes were isolated from dyeing wastewater collected at Dongducheon and Banweol industrial complex. Five microbes showed excellent ability of color removal and were identified as Shewanella putrefaciens, Aeromonas salmonicida(3 different strains), and Pseudomonas vesicularis. Five identified strains had optimal pH and optimal temperature as 7.0 and 30$^{\circ}C$ for cultivation, and showed morphological characteristics of Gram negative, oxidase negative, rod shape, and non-motility, but their biochemical characteristics were distinguishable. Each single strain of five microbes were tested in the 500 mL flask to treat dyeing wastewater, and achieved about 35% color removal efficiency in average. When two strains were selected and applied to the treatment at same time, color removal efficiency was increased up to 65%. While three or more associations of each strain did not show the improvement of color removal. Inhibition effects by $Mn^{2+}\;and\;Fe^{3+}$ on the dye degradation were tested and resulted in no effect under 70 ppm concentration.

• Journal of Soil and Groundwater Environment
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• v.16 no.6
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• pp.58-65
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• 2011

The bacterial uranium(VI) reduction and its resultant low solubility make this process an attractive option for removing U from groundwater. An impact of aqueous suspending iron phase, which is redox sensitive and ubiquitous in subsurface groundwater, on the U(VI) bioreduction by Shewanella putrefaciens CN32 was investigated. In our batch experiment, the U(VI) concentration ($5{\times}10^5M$) gradually decreased to a non-detectable level during the microbial respiration. However, when Fe(III) phase was suspended in solution, bioreduction of U(VI) was significantly suppressed due to a preferred reduction of Fe(III) instead of U(VI). This shows that the suspending amorphous Fe(III) phase can be a strong inhibitor to the U(VI) bioreduction. On the contrary, when iron was present as a soluble Fe(II) in the solution, the U(VI) removal was largely enhanced. The microbially-catalyzed U(VI) reduction resulted in an accumulation of solid-type U particles in and around the cells. Electron elemental investigations for the precipitates show that some background cations such as Ca and P were favorably coprecipitated with U. This implies that aqueous U tends to be stabilized by complexing with Ca or P ions, which easily diffuse and coprecipitate with U in and around the microbial cell.