• Title/Summary/Keyword: Alcaligenes sp.

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Molecular Detection of Catabolic Genes for Polycyclic Aromatic Hydrocarbons in the Reed Rhizosphere of Sunchon Bay

  • Kahng Hyung-Yeel;Oh Kye-Heon
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.572-576
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    • 2005
  • This study focused on detecting catabolic genes for polycyclic aromatic hydrocarbons (PAHs) distributed in the reed rhizosphere of Sunchon Bay, Korea. These marsh and mud environments were severely affected by human activities, including agriculture and fisheries. Our previous study on microbial roles in natural decontamination displayed the possibility that PAH-degrading bacteria, such as Achromobacter sp., Alcaligenes sp., Burkholderia sp. and Pseudomonas sp. play an important decontamination role in a reed rhizosphere. In order to gain further fundamental knowledge on the natural decontamination process, catabolic genes for PAH metabolism were investigated through PCR amplification of dioxygenase genes using soil genomic DNA and sequencing. Comparative analysis of predicted amino acid sequences from 50 randomly selected dioxygenase clones capable of hydroxylating inactivated aromatic nuclei indicated that these were divided into three groups, two of which might be originated from PAH-degrading bacteria. Amino acid sequences of each dioxygenase clone were a part of the genes encoding enzymes for initial catabolism of naphthalene, phenanthrene, or pyrene that might be originated from bacteria in the reed rhizosphere of Sunchon Bay.

Isolation and Characterization of A Thiodiglycol-Degrading Cupravidus sp. (Thiodiglycol를 분해하는 Cupriavidus sp.의 분리와 특성)

  • Park, Jong-Deok;Kim, Jee-Cheon;Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.311-316
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    • 2007
  • A Gram-negative bacterium capable of degrading thiodiglycol (TDG), main hydrolysis product of sulfur mustard, was isolated from ginseng field in enrichment medium supplemented with TDG as carbon source. The isolate, WS-32, grew optimally at $30-37^{\circ}C$ and pH 6.0-8.0. It was found to be similar to the genus Cupriavi연 on the basis of 165 rRNA sequence, while its biochemical properties were highly homologous to Alcaligenes faecalis. The cell growth of WS-32 strain was slightly inhibited on LB broth by TDG, but the maximum level of its growth was maintained stably in the presence of TDG. After incubation of inoculated LB medium or uninoculated LB medium containing TDG for 2 days, TDG amount of the culture filtrate was analyzed to decrease noticeably by HPLC. TDG and alcohols were also oxidized by cell-free extract of the isolate with maximum activities at pH 8.0 and $45^{\circ}C$.

Characterization of TCE-Degrading Bacteria and Their Application to Wastewater Treatment

  • Lee, Wan-Seok;Park, Chan-Sun;Kim, Jang-Eok;Yoon, Byung-Dae;Oh, Hee-Mock
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.569-575
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    • 2002
  • Two bacterial strains capable of degrading trichloroethylene (TCE), isolated form soils contaminated with various chlorinated alkenes, were identified as Alcaligenes odorous N6 and Nocardia sp. Hl7. In addition, four KCTC strains, including three strains of Pseudomonas putida and one strain of Sphingomonas chlorophenolica, exhibited an ability to degrade toluene. A. odorans N6 and Nocardia sp. H17 degraded 84% of the initial amount of TCE in a basal salts medium (BSM), containing 0.2 mM TCE as the sole source of carbon and energy, in a day. The optimal pH for growth was within a range of 7.0-8.0. A mixed culture of the four toluene-degrading isolates degraded 95% of 0.2 mM TCE with 1.5 mM toluene as an inducer, whereas no TCE was degraded by the same mixture without an inducer. When a mixed culture of all 6 isolates was used, the degradation efficiency of 0.2 mM TCE was 72% without an inducer, in a day, and 82% with toluene as an inducer. In a continuous treatment, 1,000 mg/1 of TCE in an artificial wastewater was completely removed within 18 h when an activated sludge was used along with the microbial mixture, which was 27 h laster than when only an activated sludge was used. Accordingly, it would appear that such a microbial mixture could be effectively applied to the biological treatment of wastewater containing TCE with or without an inducer.

Agrobacterium sp. ATCC 31750의 고농도 세포배양

  • Jang, Jeong-Gyun;Cha, Wol-Seok;Gang, Si-Hyeong;Park, Jae-Eok;Lee, Jung-Heon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.245-246
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    • 2000
  • Agrobacterium sp. ATCC 31750( formerly Alcaligenes faecalis subsp myxogenes) was used to produce curdlan. Since the curdlan is secondary metabolite, it is important for curdlan production to increase cell concentration. The fedbatch operation was used to increase cell concentration with addition of carbon and nitrogen sources. When the initial sucrose concentration was 20g/L, it was consumed in 24 hrs and the cell concentration was 6g/L in a batch culture. The sucrose solution(200g/L) was fed to control the sucrose concentration above 10g/L.

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Isolation of the Biodegradable Peptide Polymer-Producing Bacterial Strain and Characterization of the Polymer Produced by This Strain (Peptide계 생분해성 고분자 생산균주의 분리 및 생성 고분자의 특성)

  • 이신영;강태수김갑수유주현
    • KSBB Journal
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    • v.8 no.3
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    • pp.209-216
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    • 1993
  • For the production of biodegradable polymers from microorganisms a bacterial strain producing a biopolymer was isolated from soil. The bacteriological characteristics of this strain and physicochemical Properties of the biopolymer produced were investigated. The bacterial strain was identified as an alkalophilic Alcaligenes sp. The Purified biopolymer treated with cetylpridinium chloride and acetone was identified as an acidic biopolymer having carboxyl groups and showed strong UV absorbance (at 210nm). The biopolymer was composed of 100% glutamic acid and glutamic acid existed as $\gamma$-polyglutamic acid($\gamma$-PGA) in the form of the $\gamma$-peptide bond. The equivalent weight of this $\gamma$-PGA was estimated about 350, indicating that one acidic fraction per 2.7 residue of $\gamma$-polyglutamic acid existed. The molecular weight was $6.5{\times}10^5$ Daltons.

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Characterization of the Bacteriocin Produced by a Leuconostoc mesenteroides Strain Inhibiting the Growth of Lactobacillus sakei (Lactobacillus sakei 생육저해활성 보유 Leuconostoc mesenteroides가 생산하는 Bacteriocin의 특성)

  • Lee, Kwang-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.390-396
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    • 2011
  • Lactobacillus sakei is known to be the most populous lactic acid bacteria in over-ripened kimchi. Twenty three strains of Leuconostoc species inhibiting the growth of Lb. sakei were isolated from kimchi and amongst these the Leuconostoc mesenteroides strain CK0122 exhibited the highest antagonistic activity against Lb. sakei. The culture supernatant of the strain CK0122 was fractionated by a molecular weight cutter and lyophilized. The fraction with a molecular weight of less than 3,000 Da showed antagonistic activity against Lb. sakei. The antibacterial activity of the active fraction was sensitive to proteinase K treatment, confirming its proteinaceous nature (bacteriocin). The crude bacteriocin was active in the pH range of 4 to 7 and extremely stable after 15 min of heat treatment at $121^{\circ}C$. The crude bacteriocin inhibited the growth of Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Alcaligenes xylosoxydans, Flavobacterium sp., and Salmonella typhimurium.

Induction by Carvone of the Polychlorinated Biphenyl (PCB)-Degradative Pathway in Alcaligenes eutrophus H850 and Its Molecular Monitoring

  • Park, Young-In;So, Jae-Seong;Koh, Sung-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.804-810
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    • 1999
  • There is a possibility that carvone, a monoterpene from spearmint (Mentha spicata), could induce the bph degradative pathway and genes in Alcaligenes eutrophus H850, which is a known Gram-negative PCB degrader with a broad substrate specificity that was thoroughly investigated with Arthrobacter sp. BIB, a Gram-positive PCB degrader. The strains BIB and H850 were unable to utilize and grow on the plant terpene [(R)-(-)-carvone] (50ppm) to be recognized as a sole carbon source. Nevertheless, the carvone did induce 2,3-dihydroxybiphenyl 1,2-dioxygenase (encoded by bphC) in the strain B lB, as observed by a resting cell assay that monitors accumulation of a yellow meta ring fission product from 4,4'-dichlorobiphenyl (DCBp). The monoterpene, however, did not appear to induce the meta cleavage pathway in the strain H850. Instead, an assumption was made that the strain might be using an alternative pathway, probably the ortho-cleavage pathway. A reverse transcription (RT)-PCR system, utilizing primers designed from a conserved region of the bphC gene of Arthrobacter sp. M5, was employed to verify the occurrence of the alternative pathway. A successful amplification (182bp) of mRNA transcribed from the N-terminal region of the bphC gene was accomplished in H850 cells induced by carvone (50ppm) as well as in biphenyl-growth cells. It is, therefore, likely that H850 possesses a specific PCB degradation pathway and hence a different substrate specificity compared with B1B. This study will contribute to an elucidation of the dynamic aspects of PCB bioremediation in terms of roles played by PCB degraders and plant terpenes as natural inducer substrates that are ubiquitous and environmentally compatible.

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Bacterial Community and Biological Nitrate Removal: Comparisons of Autotrophic and Heterotrophic Reactors for Denitrification with Raw Sewage

  • Lee, Han-Woong;Park, Yong-Keun;Choi, Eui-So;Lee, Jin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1826-1835
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    • 2008
  • An autotrophic denitrification reactor (ADR-l) and a heterotrophic denitrification reactor (HDR-2) were operated to remove nitrate and nitrite in an anoxic environment in raw sewage. The $NO_3$-N removal rate of ADR-l was shown to range from 52.8% to 78.7%, which was higher than the $NO_3$-N removal rate of HDR-2. Specific denitrification rates (SDNR) of ADR-l and HDR-2 were 3.0 to 4.0 and 1.1 to $1.2\;mgNO_3$-N/gVSS/h, respectively. From results of restriction fragment length polymorphism (RFLP) of the 16S rRNA gene, Aquaspirillum metamorphum, Alcaligenes defragrans, and Azoarcus sp. were $\beta$-Proteobacteria that are affiliated with denitritying bacteria in the ADR-l. Specifically, Thiobacillus denitrificans was detected as an autotrophic denitrification bacteria. In HDR-2, the $\beta$-Proteobacteria such as Denitritying-Fe-oxidizing bacteria, Alcaligenes defragrans, Acidovorax sp., Azoarcus denitrificans, and Aquaspirillum metamorphum were the main bacteria related to denitrifying bacteria. The $\beta$-and $\alpha$-Proteobacteria were the important bacterial groups in ADR-l, whereas the $\beta$-Proteobacteria were the main bacterial group in HDR-2 based on results of fluorescent in situ hybridization (FISH). The number of Thiobacillus denitrificans increased in ADR-l during the operation period but not in HRD-2. Overall, the data presented here demonstrate that many heterotrophic denitritying bacteria coexisted with autotrophic denitrifying bacteria such as Thiobacillus denitrificans for nitrate removal in ADR-l. On the other hand, only heterotrophic denitritying bacteria were identified as dominant bacterial groups in HDR-2. Our research may provide a foundation for the complete nitrate removal in raw sewage of low-COD concentration under anoxic condition without any external organic carbon or the requirement of post-treatment.

Production of Extracellular Water Insoluble ${\beta}-1,3-Glucan$ (Curdlan) from Bacillus sp. SNC07

  • Gummadi, Sathyanarayana N.;Kumar, Kislay
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.546-551
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    • 2005
  • [ ${\beta}-1,3-Glucan$ ] (curdlan) is a water-insoluble polysaccharide composed exclusively of ${\beta}-1,3\;linked$ glucose residues. Extracellular curdlan was mostly synthesized by Agrobacterium species and Alcaligenes faecalis under nitrogen-limiting conditions. In this study, we screened the microorganisms capable of producing extracellular curdlan from soil samples. For the first time, we reported Gram-positive bacterium Bacillus sp. SNC 107 capable of producing extracellular curdlan in appreciable amounts. The effect of different carbon sources on curdlan production was studied and found that the yield of curdlan was more when glucose was used as carbon source. It was also found that maximum production was achieved when the initial concentration of ammonium and phosphate in the medium was 0.5 and 1.9 g/L respectively. In this study the curdlan production was increased from 3 to 7g/L in shake flask cultures.

Cloning of Catechol 2,3-dioxygenase Gene from Pseudomonas putida (Pseudomonas putida의 Catechol 2,3-dioxygenase 유전자의 클로닝)

  • 김영수;최봉수;민경락;김치경
    • Korean Journal of Microbiology
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    • v.29 no.3
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    • pp.155-159
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    • 1991
  • Four strains of Pseudomonas putida (NAH), Pseudomonas sp.(TOL), Achromobacter xylosoxidans, and Alcaligenes sp. were compared with their degradative capability of aromatic compounds. All of the bacterial strains were utilized catechol as a sole carbon source for growth, but signigicantly different in degradative properties for 5 other aromatic compounds. Catechol 2, 3-dioxygenase gene from P. putida (NAH) has been cloned and expressed in E. coli. The DNA clone designated pCNU101 contains NAH-derived 6 Kb insert and its physical map was characterized. A subclone (pCNU106) for the catechol dioxygenase gene in pCNU101 contained 2.0kb-DNA insery fragmented by HpaI and ClaI.

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