• Title/Summary/Keyword: Shewanella

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Eicosapentaenoic Acid (EPA) Biosynthetic Gene Cluster of Shewanella oneidensis MR-1: Cloning, Heterologous Expression, and Effects of Temperature and Glucose on the Production of EPA in Escherichia coli

  • Lee, Su-Jin;Jeong, Young-Su;Kim, Dong-Uk;Seo, Jeong-Woo;Hur, Byung-Ki
    • 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.

Mobilization of Heavy Metals in Contaminated Soils induced by Bioaugmentation of Shewanella xiamenensis HM14

  • Walpola, Buddhi Charana;Arunakumara, K.K.I.U.;Song, Jun-Seob;Lee, Chan-Jung;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.4
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    • pp.290-298
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    • 2014
  • A bacterial strain with the potential ability to solubilize heavy metals was isolated from heavy metal contaminated soils collected from abandoned mines of Boryeong area in South Korea. The bacterial strain with the highest degree of metal resistance was shown to have close proximity with Shewanella xiamenensis FJ589031, according to 16S rRNA sequence analysis, and selected for investigating the mobilization of metals in soil or plant by the strain. The strain was found to be capable of solubilizing metals both in the absence and in the presence of metals (Co, Pb and Cd). Metal mobilization potential of the strain was assessed in a batch experiment and the results showed that inoculation could increase the concentrations of water soluble Co, Pb and Cd by 48, 34 and 20% respectively, compared with those of non-inoculated soils. Bacterial-assisted growth promotion and metal uptake in sunflower (Helianthus annuus) was evaluated in a pot experiment. In comparison with non-inoculated seedlings, the inoculation led to increase the growth of H. annuus by 24, 18 and 16% respectively in Co, Pb and Cd contaminated soils. Moreover, enhanced accumulation of Co, Pb and Cd in the shoot and root systems was observed in inoculated plants, where metal translocation from root to the above-ground tissues was also found to be enhanced by the strain. Plant growth promotion and metal mobilizing potential of the strain suggest that the strain could effectively be employed in enhancing phytoextraction of Co, Pb and Cd from contaminated soils.

Reduction of Hexavalent Chromium by Shewanella sp. HN-41 in the Presence of Ferric-Citrate (구연산철 환원 조건하에서 Shewanella sp. HN-41에 의한 6가 크롬의 환원)

  • Hyemin Park;Jin-Hyeob Kwak;Ji-Hoon Lee
    • Korean Journal of Environmental Agriculture
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    • v.42 no.3
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    • pp.253-258
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    • 2023
  • In the environment, chromium often exists in a highly mobile and toxic form of Cr(VI). Therefore, the reduction of Cr(VI) to less toxic Cr(III) is considered an effective remediation strategy for Cr(VI)-contamination. In this study, the biological reduction of hexavalent chromium was examined at the concentrations of 0.01 mM, 0.1 mM, and 1 mM Cr(VI) by the dissimilatory metal-reducing bacterium, Shewanella sp. HN-41 in the presence of ferric-citrate. With the relatively condensed cell densities, the aqueous phase Cr(VI) was reduced at the proportions of 42%, 23%, and 31%, respectively for the 0.01 mM, 0.1 mM, and 1 mM Cr(VI) incubations, while Fe(III)-citrate was reduced at 95%, 88%, and 73%, respectively. Although the strain HN-41 was not considered to reduce Cr(VI) as the sole electron acceptor for anaerobic metabolism in the preliminary experiment, it has been presumed that outer-membrane c-type cytochromes such as MtrC and OmcA reduced Cr(VI) in the presence of ferric-citrate as the electron acceptor. Since this study indicated the potential of relatively high cell density for Cr(VI) reduction, it might propose a bioremediation strategy for Cr(VI) removal from contaminated waters using engineered systems such as bioreactors employing high cell growths.

Isolation and Identification of a Histamine-degrading Barteria from Salted Mackerel (자반고등어에서 histamine 분해능을 가진 세균의 분리 동정)

  • Hwang Su-Jung;Kim Young-Man
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.743-748
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    • 2005
  • Histamine can be produced at early spoilage stage through decarboxylation of histidine in red-flesh fish by Proteus morganii, Hafnia alvei or Klebsiella pneumoniae. Allergic food poisoning is resulted from the histamine produced when the freshness of Mackerel degrades. Conversely it has been reported that there are bacteria which decompose histamine at the later stage. We isolated histamine decomposers from salted mackerel and studied the characteristics to help establish hygienic measure to prevent outbreak of salted mackerel food poisoning. All the samples were purchased through local supermarket. Histamine decomposers were isolated using restriction medium using histamine 10 species were selected. Identification of these isolates were carried out by the comparison of 16S rDNA partial sequence; as a result, we identified Pseudomonas putida strain RA2 and Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudomonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia with homology of $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}and{\;}100\%$respectively. Turbidometry determination method and enzymic method were employed to determine the ability of histamine decomposition. Among those species Shewanella massilia showed the highest in ability of histamine decomposition. From these results we confirmed various histamine decomposer were present in salted mackerel product in the market.

The Growth and EPA Synthesis of Shewanella oneidensis MR-1 and Expectation of EPA Biosynthetic Pathway

  • Jeong, Young-Su;Song, Sang-Kyu;Lee, Su-Jin;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.127-133
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    • 2006
  • Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

Biodegradation Characteristics of the Eco-friendly Plastics by Seawater Microbes (해수 미생물의 환경친화성 플라스틱의 생분해 특성)

  • Kim, Mal-Nam;Yoon, Moon-Gyung
    • Korean Journal of Environmental Biology
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    • v.26 no.3
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    • pp.247-251
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    • 2008
  • Degradation behavior in the seawater of Tongyeong, Incheon, Kunsan and Hongsung was explored for Mater-Bi$^{(R)}$, poly(3-hydroxybutyrate-co-3-hydroxy valerate) (PHBV), poly(butylene succinate-co-butylene adipate) (PBSA) and polycaprolactone (PCL) which can eventually be used for various fishery tools. Acinetobacter lwoffu/junii and Shewanella algae/putrefaciens inhabited all the seawater samples. Eikenella corrodens was also detected in all the seawater samples, although identified with poor confidence by VITEK system. Mater-Bi$^{(R)}$ was degraded faster than PHBV, PBSA and PCL in the seawater in contradiction to the degradation behavior in soil environment. The seawater retrieved from Incheon exhibited the most elevated activity for the plastic degradation, which may be partly ascribed to the largest number of total viable counts.

Growth Properties of the Iron-reducing Bacteria, Shewanella putrefaciens IR-1 and MR-1 Coupling to Reduction of Fe(III) to Fe(II)

  • Park, Doo-Hyun;Kim, Byung-Hong
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.273-278
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    • 2001
  • Shewanela, putrefaciene IR-1 and MR-1 were cultivated by using various combinations electron donor-acceptor, lactate-Fe(III) lactate-nitrate, pyruvate-FE(III), pyruvate-nitrate H$_2$ acetate-Fe(III) and H$_2$-acetate-nitrate. Both strains grew fermentatively on pyruvate and lactate but not on without and electron acceptor. In culture with Fe(III), both astrains grew on pyruvate and lactate but on H$_2$-acetate- CO$_2$. In cultivation with nitrate, both stains grew on pyruvate lactage and on H$_2$-acetate-CO$_2$ The growth yields of IR-1 pyruvate, pyruvate-Fe(III) and lactate-Fe(III) were about 3.4, 3.5, and 3.6(g cell/M substrate), respectively. From the growth properties of both strains on media with Fe(III) as an electron acceptor, the bacterial growth was confirmed not to be increased by addition of Fee(III) as an electron acceptor to the growth medium, which indicates a possibility that the dissimilatory reduction of Fe(III) to Fe(III) may not be coupled to free energy production.

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Effect of Non-indigenous Bacterial Introductions on Rhizosphere Microbial Community

  • Nogrado, Kathyleen;Ha, Gwang-Su;Yang, Hee-Jong;Lee, Ji-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.40 no.3
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    • pp.194-202
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    • 2021
  • BACKGROUND: Towards achievement of sustainable agriculture, using microbial inoculants may present promising alternatives without adverse environmental effects; however, there are challenging issues that should be addressed in terms of effectiveness and ecology. Viability and stability of the bacterial inoculants would be one of the major issues in effectiveness of microbial pesticide uses, and the changes within the indigenous microbial communities by the inoculants would be an important factor influencing soil ecology. Here we investigated the stability of the introduced bacterial strains in the soils planted with barley and its effect on the diversity shifts of the rhizosphere soil bacteria. METHODS AND RESULTS: Two different types of bacterial strains of Bacillus thuringiensis and Shewanella oneidensis MR-1 were inoculated to the soils planted with barley. To monitor the stability of the inoculated bacterial strains, genes specific to the strains (XRE and mtrA) were quantified by qPCR. In addition, bacterial community analyses were performed using v3-v4 regions of 16S rRNA gene sequences from the barley rhizosphere soils, which were analyzed using Illumina MiSeq system and Mothur. Alpha- and beta-diversity analyses indicated that the inoculated rhizosphere soils were grouped apart from the uninoculated soil, and plant growth also may have affected the soil bacterial diversity. CONCLUSION: Regardless of the survival of the introduced non-native microbes, non-indigenous bacteria may influence the soil microbial community and diversity.