• Korean Journal of Microbiology
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• v.41 no.1
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• pp.13-17
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• 2005

The aim of this work was to investigate the N-terminal amino acid sequence of catechol 2,3-dioxygenase isolated from Delftia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O from strain JK-2 was $^1MGVMRIGHASLKVMDMDAAVRHYENV^{26}$, and exhibited high sequence similarity with that of C2,3O from Pseudomonas sp., Comamonas sp. JS765, Comamonas test-osteroni, or Burkholderia sp. RP007. Approximately 950-bp C2,3O was obtained through PCR using the primers derived from N-terminal amino acid sequence. Analysis of the DNA sequence revealed that the deduced 296 amino acid sequences were determined, and it showed $100\%$ identity with C2,3O from Pseudomonas sp. AW-2 and $97\%$ similarity with Comamonas sp. JS765.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.1-7
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• 2003

The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.104-111
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• 2007

In this study, aniline dioxygenase genes responsible for initial catabolism of aniline in Burkholderia sp. HY1 and Delftia sp. HY99 were cloned and the amino acid sequences were comparatively analyzed, which already have been reported as bacteria utilizing aniline as a sole source of carbon and nitrogen, B. sp. HY1 was found to have at least a plasmid, and the plasmld-cured strain, B. sp. HY1-PC obtained using mitomycin C was tested with wild type strain to investigate whether the former maintained the degradability for aniline. This proved that the aniline oxygenase gene from B. sp. HY1 was located in chromosomal DNA, not in plasmid DNA. Aniline dioxygenase small subunits from B. sp. HY1 and D. sp. HY99 were found, based on 146 amino acids, to share 79% similarity. Notably, ado2 genes from B. sp. HY1 and D. sp. HY99 which were found to be terminal dioxygenase of aniline dioxygenase small subunit showed 99% similarity in the deduced amino acid sequences with tdnA2 of Frateuria sp. ANA-18 and danA2 of D. sp. AN3, respectively. Besides, enzyme assay and amino acid sequence analysis of catechol dioxygenase supported the previous report that B. sp. HY1 might occupy ortho-cleavage pathway using catechol 1,2-dioxygenase, while D. sp. HY99 might occupy catechol 2,3-dioxygenase for meta-cleavage pathway.

• Journal of Environmental Health Sciences
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• v.35 no.5
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• pp.393-401
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• 2009

We isolated and identified a microorganism which was excellent for ammonia oxidation in the biological control of ammonia gas in odor producing materials from organic composting. The isolated strain was tested for growth characteristics and ammonia elimination efficiency under various conditions of temperature, pH, carbon concentration and ammonia concentration. The strain was isolated from a culture broth used in a $NO_2$ producing test with Griess-Ilosvay reagent. The results of 16S rRNA sequence from the isolated strain by using BLANST (Basic Local Alignment Search Tool) and confirming RDP (Ribosomal Database Project II) and ERRD (The European Ribosomal RNA Database) indicate that the strain is related to Delftia sp. UV-Spectrophotometer (Shimadzu, UVmini-1240) was used as a microbial growth test by measuring turbidity on OD660nm and ammonia concentration was measured by Spectrophotometer (HACH, DR-4000). The optimum growth culture conditions of the ammonia oxidizer Delftia sp. were $30^{\circ}C$, pH 7, glucose concentration 1.00% and $(NH_4)_2SO_4$ 0.5 g/l. Ammonia elimination efficiency was over 94% under the same conditions.

• Korean Journal of Microbiology
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• v.36 no.2
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• pp.79-83
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• 2000

Activated sludge samples were collected from a municipal sewage treatment plant and used for enrichment of microbial consortia with aniline as the sole carbon and nitrogen source. Threc aniline-degrading bacteria were obtained lrom microbial consortia and an isolate which has excellent aniline degradability was selected for this study. The isolate was Gram-negative, and identified and designated as Delfha sp. JK-2 on the basis of various physiological and biochemical tests. 10 mM aniline was completely degraded within 24 hours after inoculation of the culture. Ammonium ion was liberated in the medium transiently during the incubation and disappeared when aniline was completely degraded. Addition of glucose as a supplementary source to aniline minimal media showed significant decrease in aniline degradat~on rate for the strain Effective degradation of aniline was achieved by the addition of 0.5% nitrate as a nitrogen source, and resulted in approximately 80% higher aniline degradation compared to the absence of nitrate. Phylogenetic analysis based on 16s [DNA sequence revealed that the strain was closely related to De@ia acidovorans, with 96% overall similarity. The 16s [DNA sequence of JK-2 was also found to be closely related to those of six other clonal types, including Acidovoru, Aquaspirillum. Xylophilus, Variovorm, and Rhodofernr.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1734-1739
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• 2006

The degradation of perchloroethylene (PCE) was investigated with the serial treatment of biological reaction after dechlorination using Fenton's reagent. The dechlorination of PCE was expressed using $D_m$ (dechlorination value), calculated from ${\Delta}Cl^-mol/{\Delta}PCE$ mol, and was 2.58 with 5 mM of $H_2O_2$ and $Fe^{3+}$. The $150{\mu}M$ of PCE was transformed to $37{\mu}M$ of dichloroacetic acid (DCAA). Biological treatment with Delftia sp. N6 was applied after degradation of PCE by the Fenton reaction. The optical densities indicating cell growth were 0.53/0.10 with/without the Fenton reaction after one day, respectively. The N6 strain degraded 95% of the DCAA produced from PCE by the Fenton reaction within one day. Consequently, it seemed that the serial treatment of a Fenton reaction and biological reaction was effective in the removal of not only PCE, but also DCAA, one of the major metabolites of PCE.

• KSBB Journal
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• v.19 no.1
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• pp.50-56
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• 2004

The aim of this work was to investigate the purification and characterization dixoygenases isolated from Delftia sp. JK-2, which could utilize aniline, benzoate, and p-hydroxybenoate as sole carbon and energy source. Catechol 1,2-dioxygenase (C1, 2O), catechol 2,3-dioxygenase(C2, 3O), and protocatechuate 4,5-dioxygenase(4,5-PCD) were isolated by benzoate, aniline, and p-hydroxybenzoate. In initial experiments, several characteristics of C1 ,2O, C2, 3O, and 4,5-PCD separated with ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose were investigated. Specific activity of C1 ,2O, C2, 3O, and 4,5-PCD were approximately 3.3 unit/mg, 4.7 unit/mg, and 2.0 unit/mg. C1 ,2O and C2, 3O demonstrated their enzyme activities to other substrates, catechol and 4-methylcatechol. 4,5-PCD showed the specific activity to the only substrate, protocatechuate, but the substrates(e.g., catechol, 3-methylcatechol, 4-methylcatechol, 4-chlorocatechol, 4-nitrocatechol) did not show any specific activities in this work. The optimum temperature of C1, 2O, C2, 3O, and 4,5-PCD were 30$^{\circ}C$, and the optimal pHs were approximately 8, 8, and 7, respectively. Ag$\^$+/, Hg$\^$+/, Cu$\^$2+/ showed inhibitory effect on the activity of C1, 2O and C2, 3O, but Ag$\^$+/, Hg$\^$+/, Cu$\^$2+/, Fe$\^$3+/ showed inhibitory effect on the activity of 4,5-PCD. Molecular weight of the C1, 2O, C2, 3O, and 4,5-PCD were determined to approximately 60 kDa,35 kDa, and 62 kDa by SDS-PAGE.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.12
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• pp.1094-1101
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• 2010

Microbial fuel cells (MFC) were enriched using sediment Nakdong river, Hoidong river and protected water area in Gijang. The microbial community of sediment and enriched MFC was analyzed by FISH (fluorescent in situ hybridization) and 16S rDNA sequencing. ${\alpha}$-Proteobacteria, Acidobacter and Cyanobactia group were dominant in sediment by FISH. The coulombs of the final 10 peak of the 3 MFC (Nakdong, Hoidong, Gijang) were 0.64 C, 0.50 C, 0.61 C, respectively. When MFCs were enriched by sediment, ${\beta}$-, ${\gamma}$-Proteobacteria, Acidobacter and Firmicutes group increased 45~90%, 50~90%, 40~80% and 45~125%, respectively. In results of 16S rDNA sequencing, Roseomonas sp., Azospillium sp., Frateuria sp., Dyella sp., Enterobacter sp. and Deinocossus were isolated from Nakdong river and Azospillium sp., Delftia sp., Ralstonia sp., Klebsiella sp. and Deinococcus sp. were isolated from protected water area in Gijang and Pseudomonas sp., Klebsiella sp., Deinococcus sp., Leifsonia sp. and Bacillus sp. were isolated from Hoidong river.

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.180-184
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• 2006

Nineteen nitrogen-fixing bacteria were isolated from rice roots cultivated in the southern part of the Korean peninsula. Among them, three isolates - Stenotrophomonas sp. KNUC89, Pseudomonas sp. KNUC116, and Delftia sp. KNUC133 - showed antifungal activities against phytopathogenic fungi Rhizoctonia solani and Fusarium oxysporum. Besides they could produce auxin and siderophores.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.378-384
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• 2005

Vanillin is one of the world's principal flavoring compounds, and is used extensively in the food industry. The potential vanillin production of the bacteria was compared to select and clone genes which were appropriate for highly productive vanillin production by E. coli. The fcs (feruloyl-CoA synthetase) and ech (enoyl-CoA hydratase/aldolase) genes cloned from Amycolatopsis sp. strain HR104 and Delftia acidovorans were introduced to pBAD24 vector with $P_{BAD}$ promoter and were named pDAHEF and pDDAEF, respectively. We observed 160 mg/L vanillin production with E. coli harboring pDAHEF, whereas 10 mg/L of vanillin was observed with pDAHEF. Vanillin production was optimized with E. coli harboring pDAHEF. Induction of the fcs and ech genes from pDAHEF was optimized with the addition of 13.3 mM arabinose at 18 h of culture, from which 450 mg/L of vanillin was produced. The feeding time and concentration of ferulic acid were also optimized by the supplementation of $0.2\%$ ferulic acid at 18 h of culture, from which 500 mg/L of vanillin was obtained. Under the above optimized condition of arabinose induction and ferulic acid supplementation, vanillin production was carried out with four different types of media, M9, LB, 2YT, and TB. The highest vanillin production, 580 mg/L, was obtained with LB medium, a 3.6 fold increase in comparison to the 160 mg/L obtained before the optimization of vanillin production.