• Title/Summary/Keyword: Rhodococcus rhodochrous

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Investigation about enzymatic properties of the gene encoding catechol 1,2-dioxygenase from Phenol-degrading, Rhodococcus sp. EL-GT

  • 이희정;이오미;김기한;박근태;박재림;이상준
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2001.11a
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    • pp.102-104
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    • 2001
  • 본 연구는 방향족 화합물질 중 페놀폐수에 대한 생물학적 처리를 위해 본 실험실에서 분리한 페놀분해능이 우수한 Rhodococcus sp. EL-GT를 이용하여 catechol 분해 catechol 1,2-dioxygenase 분해활성을 측정하였고, 이것이 ortho-pathway임을 확인할 수 있었다. 또한 다른 연구에서 보고된 Rhodococrus rhodochrous NCIMB 13259 균주의 catechol 1,2 dioxygenase를 기초로한 primer를 이용하여 PCR을 수행하였으며 이 분해 유전자의 cloning실험을 수행 중이다. 이들 실험을 통하여 Rhodoroccus sp. EL-GT의 페놀분해 균의 유전적 구조 및 특성을 검토하고 이를 이용하여 방향족 화합물의 분해능이 보다 우수한 균주의 개발을 시도하고자 한다.

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Production of 3-Hydroxypropionic Acid from Acrylic Acid by Newly Isolated Rhodococcus erythropolis LG12

  • Lee, Sang-Hyun;Park, Si-Jae;Park, Oh-Jin;Cho, Jun-Hyeong;Rhee, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.474-481
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    • 2009
  • A novel microorganism, designated as LG12, was isolated from soil based on its ability to use acrylic acid as the sole carbon source. An electron microscopic analysis of its morphological characteristics and phylogenetic classification by 16S rRNA homology showed that the LG12 strain belongs to Rhodococcus erythropolis. R. erythropolis LG12 was able to metabolize a high concentration of acrylic acid (up to 40 g/l). In addition, R. erythropolis LG12 exhibited the highest acrylic acid-degrading activity among the tested microorganisms, including R. rhodochrous, R. equi, R. rubber, Candida rugosa, and Bacillus cereus. The effect of the culture conditions of R. erythropo/is LG12 on the production of 3-hydroxypropionic acid (3HP) from acrylic acid was also examined. To enhance the production of 3HP, acrylic acid-assimilating activity was induced by adding 1 mM acrylic acid to the culture medium when the cell density reached an $OD_{600}$ of 5. Further cultivation of R. erythropo/is LG 12 with 40 g/l of acrylic acid resulted in the production of 17.5 g/l of 3HP with a molar conversion yield of 44% and productivity of 0.22 g/l/h at $30^{\circ}C$ after 72 h.

Biodegradation of Aromatic Compounds by Nocardioform Actinomycetes

  • CHA CHANG-JUN;CERNIGLIA CARL E.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.157-163
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    • 2001
  • Mycolic acid-containing gram-positive bacteria, so called nocardioform actinomycetes, have become a great interest to environmental microbiologists due to their metabolic versatility, multidegradative capacity and potential for bioremediation of priority pollutants. For example, Rhodococcus rhodochrous N75 was able to metabolize 4-methy1catechol via a modified $\beta$-ketoadipate pathway whereby 4-methylmuconolactone methyl isomerase catalyzes the conversion of 4-methylmuconolactone to 3-methylmuconolactone in order to circumvent the accumulation of the 'dead-end' metabolite, 4-methylmuconolactone. R. rhodochrous N75 has also shown the ability to transform a range of alkyl-substituted catechols to the corresponding muconolactones. A novel 3-methylmuconolactone-CoAsynthetase was found to be involved in the degradation of 3-methylmuconolactone, which is not mediated in a manner analogous to the classical $\beta$-ketoadipate pathway but activated by the addition of CoA prior to hydrolysis of lactone ring, suggesting that the degradative pathway for methylaromatic compounds by gram-positive bacteria diverges from that of proteobacteria. Mycobacterium sp. Strain PYR-l isolated from oil-contaminated soil was capable of mineralizing various polyaromatic hydrocarbons (PAHs), such as naphthalene, phenanthrene, pyrene, fluoranthrene, 1-nitropyrene, and 6-nitrochrysene. The pathways for degradation of PAHs by this organism have been elucidated through the isolation and characterization of chemical intermediates. 2-D gel electrophoresis of PAH-induced proteins enabled the cloning of the dioxygenase system containing a dehydrogenase, the dioxygenase small ($\beta$)-subunit, and the dioxygenase large ($\alpha$)-subunit. Phylogenetic analysis showed that the large a subunit did not cluster with most of the known sequences except for three newly described a subunits of dioxygenases from Rhodococcus spp. and Nocardioides spp. 2-D gel analysis also showed that catalase-peroxidase, which was induced with pyrene, plays a role in the PAH metabolism. The survival and performance of these bacteria raised the possibility that they can be excellent candidates for bioremediation purposes.

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Environmental Factors Affecting on Shrimp Cultivation and Bacterial Examination in Shrimp Aquaculture

  • Chun, Jae-Woo;Ma, Chae-Woo;Oh, Kye-Heon
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.775-779
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    • 2003
  • Cultivation environment for effective production of shrimp in shrimp aquaculture, Institute of Marine & Fishery, Soonchunhyang University located at Tae-An, Chung-Nam Province, with the ultimate aim of probiotics development, was monitored. Several environmental factors including dissolved oxygen, water body temperature, pH, salinity, $NH_4-N$, $NO_2-N$, $NO_3-N$, chlorophyll a affecting on the growth of shrimp were examined. Concentrations of $NH_4-N$, $NO_2-N$, and $NO_3-N$ dissolved in water samples were determined to 0.024-0.034 mg/L, 0.03-0.02, and 0.004-0.009 mg/L, respectively. Chlorophyll a content was examined in the range of 0.002-0.118 $ug/m^3$. In order to understand the distribution of different bacteria in water samples collected in shrimp aquaculture, bacteria were isolated and enumerated on the marine agar plates. Total number of bacteria were increased to approximately $6.5\;{\times}\;10^4$. Thirteen predomonant bacteria were isolated and identified. As the results of BIOLOG test of the isolates, these bacteria were identified as Corynebacterium nirilophilus, Clavibacter agropyri, Sphingomonas adhaesiva, Brevundimonas vesicularis, Vibrio parahaemolyticus, Pseudomonas bathycetes, Vibrio tubiashii, Sphingomonas macrogoltabidus, Rhodococcus, rhodochrous, Burkholderia glumae, Corynebacterium urealyticus, Rhodococcus fascians, Psychrobacter immobilis, respectively. Further work will stimulate the elucidation of pathogenicity, corresponding bacteria related to environment and probiotics, providing good information for effective production of shrimp.

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Twelve previously unrecorded bacterial species, isolated from the Nakdong River, South Korea

  • Kim, Hyangmi;Han, Ji-Hye
    • Journal of Species Research
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    • v.10 no.2
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    • pp.134-141
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    • 2021
  • During a survey of indigenous prokaryotic species diversity of the upstream Nakdong River, South Korea, 12 bacterial strains were isolated for further analysis. These bacterial strains were identified showing at least 98.7% 16S rRNA gene sequence similarity with known bacterial species that were previously unreported in South Korea. The 12 bacterial strains were phylogenetically diverse and assigned to four classes, eight orders, nine families, and ten different genera. The isolates were identified as Leucobacter holotrichiae (99.1%), Leucobacter tardus (99.9%), Rhodococcus rhodochrous (99.9%), Tessaracoccus oleiagri (100%), and Paeniglutamicibacter cryotolerans (99.3%), of the class Actinobacteria; Bacillus coagulans (99.7%) and Bacillus wudalianchiensis (99.1%) of the class Bacilli; Ochrobactrum pseudogrignonense (99.2%) and Paracoccus thiocyanatus (100%) of the class Alphaproteobacteria; and Ideonella azotifigens (99.0%), Polaromonas glacialis(99.3%), and Herbaspirillum seropedicae (99.5%) of the class Betaproteobacteria. The cellular and colonial morphology, biochemical properties, and phylogenetic position of these isolates were examined, and species descriptions are provided.

Desulfurization of Dibenzothiophene and Diesel Oil by Metabolically Engineered Escherichia coli

  • Park, Si-Jae;Lee, In-Su;Chang, Yong-Keun;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.578-583
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    • 2003
  • The desulfurization genes (dszABC) were cloned from Gordonia nitida. Nucleotide sequences similarity between the dszABC genes of G. nitida and those of Rhodococcus rhodochrous IGTS8 was 89%. The similarities of deduced amino acids between the two were 86% for DszA, 86% for DszB, and 90% for DszC. The G. nitida dszABC genes were expressed in several different Escherichia coli strains under an inducible trc promoter. Cultivation of these metabolically engineered E. coli strains in the presence of 0.2 mM dibenzothiophene (DBT) allowed the conversion of DBT to 2-hydroxybiphenyl (2-HBP), which is the final metabolite of the sulfur-specific desulfurization pathway. The maximum conversion of DBT to 2-HBP was 16% in 60 h. Recombinant E. coli was applied for the deep desulfurization of diesel oil supplemented into the medium at 5% (v/v). Sulfur content in diesel oil was decreased from 250 mg sulfur/1 to 212.5 mg sulfur/1, resulting in the removal of 15% of sulfur in diesel oil in 60 h.

Cloning and Phylogenetic Analysis of Two Different bphC Genes and bphD Gene From PCB-Degrading Bacterium, Pseudomonas sp. Strain SY5

  • Na, Kyung-Su;Kim, Seong-Jun;Kubo, Motoki;Chung, Seon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.668-676
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    • 2001
  • Pseudomonas sp. strain SY5 is a PCB-degrading bacterium [24] that includes two different enzymes (BphC1 and BphC2) encoding 2,3-dihdroxybiphenyl 1,2-dioxygenase and BphD encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase. The bphC1 and bphC2 genes were found to consist of 897 based encoding 299 amino acids and 882 bases encoding 294 amino acids, respectively, whereas the bphD gene consisted of 861 bases encoding 287 amino acids. According to a homology search, a 50% and 39% similarity between the bphC1 and bphC2 genes at the nucleotide and amino acid level was shown, respectively. The bphC1 gene showed a 38% and 45% similarity at the amino acid level to Alcaligenes eutrophus A5 and Rhodococcus rhodochrous, respectively, whereas, bphC2 showed a 95% and 43% similarity, respectively. A comparison of the deduced amino acid sequence of the bphD product of Pseudomonas sp. SY5 with that of A. eutrophus A5, Pseudomons sp. KKS102, and LB400 showed a sequence identity of 92, 92, and 79%, respectively. Strain SY5 was originally isolated from municipal sewage containing recalcitrant organic compounds an found to have a high degradability of various aromatic compounds [23]. The current study found that strain SY5 had two extradiol-type dioxygenases, which did not hybridize with each other as they had a low similarity, yet a similar structure of evolutionarily conserved amino acids residues for catalytic activity between BphC1 and BphC2 was observed.

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Genetic Organization of the dhlA Gene Encoding 1,2-Dichloroethane Dechlorinase from Xanthobacter flavus UE15

  • Song, Ji-Sook;Lee, Dong-Hun;Lee, Kyoung;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.188-193
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    • 2004
  • Xanthobacter flavus strain UE15 was isolated in wastewater obtained from the Ulsan industrial complex, Korea. This strain functions as a 1,2-dichloroethane (1,2-DCA) degrader, via a mechanism of hydrolytic dechlorination, under aerobic conditions. The UE15 strain was also capable of dechlorinating other chloroaliphatics such as 2-chloroacetic acid and 2-chloropropionic acid. The dhlA gene encoding 1,2-DCA dechlorinase was cloned from the genomic DNA of the UE15 strain, and its nucleotide sequence was determined to consist of 933 base pairs. The deduced amino acid sequence of the DhlA dechlorinase exhibited 100% homology with the corresponding enzyme from X. autotrophicus GJ10, but only 27 to 29% homology with the corresponding enzymes from Rhodococcus rhodochrous, Pseudomonas pavonaceae, and Mycobacterium sp. strain GP1, which all dechlorinate haloalkane compounds. The UE15 strain has an ORF1 (1,356 bp) downstream from the dhlA gene. The OFR1 shows 99% amino acid sequence homology with the transposase reported from X. autotrophicus GJ10. The transposase gene was not found in the vicinity of the dhlA in the GJ10 strain, but rather beside the dhlB gene coding for haloacid dechlorinase. The dhlA and dhlB genes were confirmed to be located at separate chromosomal loci in the Xanthobacter flavus UE15 strain as well as in X. autotrophicus GJ10. The dhlA and transposase the UE15 strain were found to be parenthesized by a pair of insertion sequences, 181247, which were also found on both sides of the transposase gene in the GJ10 strain. This unique structure of the dhlA gene organization in X. flavus strain UE15 suggested that the dechlorinase gene, dhlA, is transferred with the help of the transposase gene.