• 제목/요약/키워드: Shewanella

검색결과 75건 처리시간 0.025초

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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    • 제11권6호
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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
    • 한국토양비료학회지
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    • 제47권4호
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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.

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

  • 박혜민;곽진협;이지훈
    • 한국환경농학회지
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    • 제42권3호
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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.

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

  • 황수정;김영만
    • 생명과학회지
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    • 제15권5호
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    • pp.743-748
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    • 2005
  • Histamine은 적색육 어류의 histidine이 어육 중의 Morganella morganii, Hafnia alvei 및 Klebsiella pneumoniae와 같은 부패세균에 의해 탈탄산 되어 초기에 형성되는 것으로 allergy성 식중독을 일으킬 수 있다. 이는 적색육 어류인 고등어의 선도저하 시에 많이 생성된다. 그리고 부패 후기에는 histamine을 분해하는 세균도 존재하는 것으로 알려져 있다. 그러므로, histamine 식중독의 잠재력을 지닌 자반고등어로 인한 식중독 사고 예방과 그 위생 대책을 수립하는데 필요한 자료를 얻고자 자반고등어에서 histamine 분해능을 가진 균을 분리, 동정하였다. 시료는 대형마트에서 시판되는 상태로 구입하였다. 질소원과 탄소원으로써 histamine만을 첨가한 제한배지를 사용하여 histamine 분해능을 가진 균을 분리하였다. 그리고 Cram staining, oxidase, catalase, citrate, TSI test, $H_{2}S$ reaction 및 indole 생성 등의 기본적인 생화학적 동정시험을 거쳐 10종의 시험균주를 선택하였다. 이 균주들을 16SrRNA gene 염기서열 비교에 의한 계통발생학적 분석을 이용하여 동정 하였다. 그 결과, Pseudomonas putida strain RA2, Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudemonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia의 10종이 모두 동정 되 었으며, 각각 $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}100\%$의 상동성을 보였다. Histamine분해능의 존재를 탁도측정법과 효소법에 의해 확인한 결과, 분리된 10종 모두의 histamine 분해능이 재확인 되었고, 그 중 Shewanella massilia가 최대의 histamine 분해능을 보이는 것으로 확인되었다. 이 결과로 자반고등어 시판 제품에는 다수의 histamine 분해 세균이 존재하는 것을 확인할 수 있었으며, 이 세균을 활용한다면 식품 내 존재하는 histamine을 효과적으로 분해할 수 있을 것이라 예상된다.

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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    • 제11권2호
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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.

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

  • 김말남;윤문경
    • 환경생물
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    • 제26권3호
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    • pp.247-251
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    • 2008
  • 통영, 인천, 군산 및 홍성의 해수 미생물에서 각종 어업용구의 재료로 사용될 수 있는 Mater-Bi$^{(R)}$, PHBV, PBSA 및 PCL의 분해거동을 조사하였다. Acinetobacter lwoffu/junii와 Shewanella algae/putrefaciens는 모든 해수속에 서식하고 있었으며 Eikenella corrodens 역시 비록 YITEK 결과의 신뢰도가 조금 낮은 수준으로 동정되었지만 모든 해수에서 검출되었다. 해수에서는 Mater-Bi$^{(R)}$가 PHBV, PBSA및 PCL보다 더 빠르게 분해되어 토양 환경에서의 분해와는 다른 거동을 나타내었다. 인천의 해수가 이들 플라스틱에 대하여 가장 높은 분해활성을 보였으며 이는 인천의 해수가 가장 많은 수의 total viable count를 포함하고 있는데 일부 기인하는 것으로 사료된다.

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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    • 제39권4호
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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
    • 한국환경농학회지
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    • 제40권3호
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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.