• 한국미생물·생명공학회지
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• 제28권4호
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• pp.202-208
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• 2000

Four bacteria capable of using aniline as a sole source of carbon and energy we4e isolated from river waters. Among them, two strains were identified as Stenotrophomonas maltophilia based on their physiological and biochemical characteristics and 16SrRNA gene sequence and the others as delftia acidovorans. The four strains were able to grow on the mineral salt media containing aniline at concentrations up to 6,000 $\mu\textrm{g}$/ml. Since aniline degradation by S. maltophilia has not been reported so far, the two strains A-s and 51-4 were selected for further studies. They completely utilized aniline in a mineral salt medium containing 300 $\mu\textrm{g}$/ml of aniline as a sole carbon and energy source within 24 hours. Optimum pH and temperature for aniline degradation and cell growth of both strains were 7.0 and $35^{\circ}C$, respectively. In addition, they effectively degraded aniline is waste, underground and river waters containing 300 $\mu\textrm{g}$/ml of aniline. This is the first report of aniline degradation by S. maltophilia strains.

• 미생물학회지
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• 제30권3호
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• pp.199-206
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• 1992

Six isolated strains degrading aniline were selected, identified and designated as pseudomonas putida K6, Pseudomonas acidovorans K82, Achromobacter gr. D. V. K24, Achromobacter xylosocidans K4, Moraxella sp. K21 and Moraxella sp. K22. All of them degraded 1000 ppm aniline completely within 30 to 36 hours. Most of these strains are resistant to antibiotics more than one, but Moraxella sp. has not any antibiotic marker tested. Most strains except for P. acidovorans K82 were shown to have resistance to the heavy metal ions such as Ni, Cu, Li, Ba, Co, etc. but not to Hg to which only P. putida K6 was resistant. M. sp. K21 was capable of degrading aniline to a maximum concentration of 2500 ppm without any repression. The incubation of the cell in limited pH ranges (4-8) had no great effect on aniline degradation. The addition of bactopeptone to the minimal media promoted the speed of aniline degradation, but the addition of glucose rather repressed the rate of aniline degradation. Through enzyme assay, A. gr. D. V. K 24 was shown to degrade aniline through artho-pathway and formed .betha.-ketoadipate as intermediate metabolite.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.643-650
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• 2000

A new aniline-utilizing microorganism, strain HY1 obtained from an orchard soil, was characterized by using the BIOLOG system, an analysis of the total cellular fatty acids, and a 16S rDNA sequence. Strain HY1 was identified as a Burkholderia species, and was designated Burkholderia sp. HY1. GC and HPLC analyses revealed that Burkholderia sp. HY1 was able to degrade aniline to produce catechol, which was subsequently converted to cis,cis-muconic acid through an ortho-ring fission pathway under aerobic conditions. Strain HY1 exhibited a drastic reduction in the rate of aniline degradation when glucose was added to the aniline media. However, the addition of peptone or nitrate to the aniline media dramatically accelerated the rate of aniline degradation. A fatty acid analysis showed that strain HY1 was able to produce lipids 16:0 2OH, and 11 methyl 18:1 ${\omega}7c$ approximately 3.7-, 2.2-, and 6-fold more, respectively, when grown on aniline media than when grown on TSA. An analysison the alignment of a 1,435 bp fragment. A phylogenetic analysis of the 16S rDNA sequence based on a 1,420 bp multi-alignment sowed of the 16s rDNA sequence revealed that strain HY1 was very closely related to Burkholderia graminis with 95% similarity based that strain HY1 was placed among three major clonal types of $\beta$-Proteobacteria, including Burkholderia graminis, Burkholderia phenazinium, and Burkholderia glathei. The sequence GAT(C or G)${\b{G}}$, which is highly conserved in several locations in the 16S rDNA gene among the major clonal type strains of $\beta$-Proteobacteria, was frequently replaced with GAT(C or G)${\b{A}}$ in the 16S rDNA sequence from strain HY1.

• Archives of Pharmacal Research
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• 제15권1호
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• pp.41-47
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• 1992

In order to obtain acetaminophen, a popular analgesic-antipyretic, though microbial p-hydroxylation and N-acetylation of aniline, various Streptomyces strains were screened. Aniline N-acetylation activity was rather ubiquitous but-hydroxylation activity was selective. Microbial conversion pathway of aniline to acetaminophen was considered to be through N-acetylation and p-hydroxylation or vice versa. However, depending on species used, o-hydroxylation and its degradation activity (S. fradiae) and acetaminophen degradation activity (S. coelicolar) were also detected. Among the screened Streptomyces strains, S fradiae NRRL 2702 showed the highest acetanilide p-hydroxylation activity (203% conversion rate). Furthermore, in S. fradiae carbon source and its concentration, phosphate ion concentration and pH of growth medium were found to play the crucial roles in p-hydroxylation activity. Through the proper combination of factors mentioned above, the ten times more activity (26-30% conversion rate) was attained.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.79-84
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• 2001

An aniline-utilizing microorganism identified as a species of Pseudomonas was isolated from soil contaminated highly with aniline and urea-herbicide. This strain was able to utilize aniline as the sole source of carbon and energy, and was shown to harbor a single large plasmid mediating the aniline assimilation. Subsequent plasmid-curing of this bacterium resulted in the abolishment of the aniline utilizing phenotype and the loss of catechol-C2,3O-oxygenase. The reestablishment of the plasmid, denoted pB10, in cured Pseudomonas sp. via filter surface mating, resulted in restoration of the aniline assimilation abilities and enzyme activity.

• 미생물학회지
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• 제36권2호
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• pp.79-83
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• 2000

도시폐수처리장의 활성슬러지 표본으로부터 유일 탄소원 및 질소원으로 aniline을 이용할수 있는 미생물 컨소시엄을 농화배양하였다. 농화배양으로부터 분리된 3개의 분해세균 가운데 분해능이 탁원한 세균을 본 연구에 사용되었다. 분리세균운 그람 음성세균으로서 여러 가지 생리.생화학적 시험을 통하여 Delftia acidovorans로 동정되었으며 이를 Delftia sp. JK-2로 명명하였다. 10mM의 aniline이 포함된 액체배지에 Delftia sp. JK-2를 접종한 결과 24시간 이내에 aniline이 완전히 분해되엇다. 이 가간동안 배양액 내에 $NH_4^+$이온은 일시적으로 생성되었다가 aniline이 완전히 분해됨에 따라 완전히 사라졌다. Aniline을 포함하여 무기배지에 부가탄소로서 glucose를 첨가하였을 때 분해능은 크게 감소하였다. 질소원으로서 0.5% nitratef,f 첨가하였을 때 첨가하지 않은 대조군에 비해 anline의 분해가 80% 이상 향상되었다. Aniline 분해세균의 16S rDNA 염기서열을 이용하여 phylogenetic 분석을 실시한 결과 이 세균은 Delftia acidovorans와 96%의 유사성을 나타내었으며, Acidovorax, Aquaspirllum, Xylophilus, Variovorax, Rhodoferax 등의 세균과도 상당한 유사성을 나타내었다.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.25-31
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• 2003

아닐린을 분해할 수 있는 Deiftia acidovoran 51-A가 기존에 분리되어 아닐린 분해능이 우수함이 보고되었다. 이 균주로부터 아닐린 분해관련 유전자를 선발하기 위하여 Tn5-B20삽입 변이체들을 유도하여 영양요구형이 아니면서 아닐린을 분해할 수 없는 변이균주 D. acidovorans 10-4-2 균주를 선발하였다. Southern hybridization 결과 이 변이균주에는 Tn5-B20이 한 copy만 삽입된 것으로 나타났다 이 변이균주의 Tn5-B20삽입의 인접 유전자들을 분리하여 염기서열을 분석한 결과 아닐린 분해의 첫 단계에 해당하는 아닐린의 catechol로의 산화적 deamination에 관련되어 있는 것으로 추정하는 tdnQ, tdnT tdnAl 유전자들이 동정되었고 Tn5-B2O은 tdnAl의 바로 밑에 삽입된 것을 알 수 있었다. TdnA2 및 downstream의 유전자 기능을 상실하여 아닐린을 catechol로 전환하는 과정에 변이가 발생하고 따라서 아닐린을 탄소원으로 이용하지 못하는 표현형을 가지게 된 것으로 결론지을 수 있었다. Tn5-B20 의 일부 DNA 조각을 probe로 Southern hybridization 결과 transposon 삽입이 대형의 플라스미드에 삽입된 것으로 나타나 tdn 유전자들이 pTDN51이라고 명명한 100-kb 이상의 대형 플라스미드에 위치함을 알 수 있었다. 이상의 결과는 D. acidovorans 51-A의 pTDN51상의 tdn유전자들이 아닐린의 분해에 관여함을 보여준다.

• 한국환경농학회지
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• 제20권2호
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• pp.74-78
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• 2001

아닐린을 유일 탄소원 및 에너지원으로 이용하는 두 개의 균주를 강물에서 분리하였다. 두 개의 세균 균주들은 생리 ${\cdot}$ 생화학적 특성과 16S rRNA 유전자 염기서열을 통하여 모두 Pseudomonas rhodesiae로 동정되었다. 이 균주들은 유일 탄소원으로 아닐린이 6,000 ${\mu}g/mL$ 수준으로 포함되어 있는 최소배지에서도 생육이 가능하였으며 두 균주간의 아닐린 분해능에는 뚜렷한 차이가 없었다. P. rhodesiae 51-C 균주는 아닐린이 300 ${\mu}g/mL$ 수준으로 처리된 최소배지에서 16시간이내에 아닐린을 완전히 분해하였고 아닐린 분해에 대한 최적의 pH는 7.0이었으며, 적온은 $30^{\circ}C$와 $35^{\circ}C$사이였다. P. rhodesiae에 의한 아닐린의 분해는 처음으로 보고하는 바이다.

• Journal of Microbiology and Biotechnology
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• 제26권5호
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• pp.891-899
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• 2016

The recently isolated aniline-degrading bacterium Pseudomonas migulae AN-1 was tagged with green fluorescent protein (GFP) to investigate its bioaugmentation potential against aniline-contaminated groundwater through microcosm experiments. The survival and cellular response of GFP-tagged AN-1 introduced in a lab-scale aquifer corresponded directly with aniline consumption. During the process, the GFP-tagged AN-1 biomass increased from 7.52 × 10⁵ cells/ml to 128 × 10⁵ cells/ml and the degradation rate of aniline was 6.04 mg/l/h. GFP-tagged AN-1 was moderately hydrophobic (41.74%-47.69%) when treated with 20-100 mg/l aniline and exhibited relatively strong hydrophobicity (55.25%-65.78%) when the concentration of aniline was ≥100 mg/l. The membrane permeability of AN-1 increased followed by a rise in aniline below 100 mg/l and was invariable with aniline above 100 mg/l. Pyrosequencing analysis showed that the relative abundance of Proteobacteria (accounted for 99.22% in the non-bioaugmentation samples) changed to 89.23% after bioaugmentation with GFP-tagged AN-1. Actinobacteria increased from 0.29% to 2.01%, whereas the abundance of Firmicutes barely changed. These combined findings demonstrate the feasibility of removing aniline in aquifers by introducing the strain AN-1 and provide valuable information on the changes in the diversity of dominant populations during bioaugmentation.

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.427-431
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• 1988

Pseudomonas testosteroni 6F1의 아닐린 분해에 미치는 이차기질의 첨가효과를 보기 위하여 균생장에 필요한 유도기간과 효소활성유도 정도를 조사하였다. 아닐린만 존재하는 배지에서 P. testosteroni 6Fl은 7시간의 유도기간을 필요로 한데 비하여 쉽게 이용가능한 이차기질을 첨가해준 경우 아닐린 분해에 필요한 유도기간이 1-3시간으로 줄어들었다. 이러한 아닐린 분해 촉진효과는 이차기질과 아닐린이 서로 발리 이용되면서 균체의 생장과 분해효소의 활성유도시기를 앞당겼기 때문이었으며, 이때 아닐린 분해효소의 최종활성 정도는 첨가해준 이차기질에 따라 다르게 나타났다.