• Journal of Applied Biological Chemistry
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• 제43권1호
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• pp.44-48
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• 2000

Burkholderia cepacia strain G4 (pHG-2) containing toluene 2-monooxygenase and toluene dioxygenase, was able to grow on toluene and accumulate cis-3-methyl-3,5-cyclohexadien-1,2-diol (cis-toluene dihydrodiol) in the liquid culture. The cis-toluene dihydrodiol produced was identical to the authentic compound, as judged through mass spectrometry and nuclear magnetic resonance analysis. Our results indicate that pHG-2 provides an economical means to produce chemically-important chiral synthons while growing on toluene.

• KSBB Journal
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• 제15권5호
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• pp.474-481
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• 2000

The effects of growth substrates such as toluene and phenol on cometabolic biodegradation of trichloroethylene (TCE) by Burkholderia cepacia G4 were investigated. The dual effects of primary substrate on TCE biodegradation, stimulatory effects of toluene and phenol at low concentrations (0.5∼2 ppm & 0.1∼0.5 ppm, respectively) and a competitive inhibition at high concentration, were observed in batch experiments. These stimulatory effects of toluene and phenol were found to be due to the increments in the amount of reducing power like NADH which could be generated during the assimilation of toluene and phenol as the carbon and energy source. The efficiency of TCE biodegradation in trickling biofilm reactor (TBR) could be also enhanced up to the TCE removal efficiency of 58.1% by the supply of appropriate amounts of phenol (0.94∼4.7 ppm).

• 한국미생물·생명공학회지
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• 제31권4호
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• pp.421-428
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• 2003

길항미생물을 이용한 항곰팡이성 퇴비의 개발을 목적으로 공시 식물로써 Spinacia oleracea L과 식물 병원균 Rhizoctonia solani Kuhn O-28을 모델로 사용하여 수행 되었다. 다양한 음식쓰레기를 일년동안 숙성시킨 퇴비로부터 80 균주를 분리하였고, 그 중 No.15-2 균주가 R. solani Kuhn O-28에 대해 가장 높은 항곰팡이 활성을 보였다. 16S rDNA sequencing과 primer pair PCR에 의해 표현형과 분류학적으로 거의 독성이 없는 것으로 밝혀진 Burkholderia cepacia genomoval V로 분류되었다. 이 B. cepacia No. 15-2가 퇴비화 중에 우점하였으며 그 균수는 15일 동안 거의 $10^{13}$ cfu/g을 유지하였다. S. oleracea L을 배양 했을 경우 R. solani Kuhn O-28에 의한 발병율은 B. cepacia No.15-2를 첨가함으로써 40% 감소하였다. 결론적으로 B. cepacia No.15-2는 다양한 식물의 곰팡이 유래의 병을 방제하는 데 이용 가능할 것으로 사료된다.

• KSBB Journal
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• 제13권5호
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• pp.561-568
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• 1998

Two cometabolic trichloroethylene (TC) degraders, Pseudomonas putida F1 and Burkholderia (Pseudomonas) cepacia G4, were found to catabolize phenol, benzene, toluene, and ethylbenzene as carbon and energy sources. Resting cells of P. putida F1 and B. cepacia G4 grown in the presence of toluene and phenol, respectively, were able to degrade not only benzene, toluene and ethylenzene but also TCE and p-xylene. However, these two strains grown in the absence of toluene or phenol did not degrade TCE and p-xylene. Therefore, it was tentatively concluded that cometabolic degradation of TC and p-xylene was mediated by toluene dioxygenase (P. putida F1) or toluene-2-monooxygenase (B. cepacia G4). Maximal degradation rates of BTEX and TCE by toluene- and phenol-induced resting cells of P. putida F1 and B. cepacia G4 were appeared to be 4-530 nmol/(min$.$mg cell protein) when a single compound was solely served as a target substrate. In case of double substrates, the benzene degradation rate by P. putida F1 in the presence of toluene was decreased up to one seventh of that for the single substrate. TCE degradation rate was also linearly decreased as toluene concentration increased. On the other hand, toluene degradation rate was enhanced by benzene and TCE. For B. cepacia G4, degradation rates of TCE and toluene increased 4 times in the presence of 50 ${\mu}$M phenol. From these results, it was concluded that a degradation rate of a compound in the presence of another cosubstrate(s) could not be predicted by simply generalizing antagonistic or synergistic interactions between substrates.

• KSBB Journal
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• 제16권5호
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• pp.511-515
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• 2001

TCE및 TCE 분해산물의 높은 독성으로 인하여 TCE 분해 효소 및 세포 불활성화가 일어나서 장기간 안정되게 반응기 운전이 어렵다는 단점을 극복하고, TCE와 성장기질사이의 경쟁적 저해관계로 인한 처리효율 감소도 막기 위하여 TCE분해단계와 세포 및 효소 재활성화 단계를 구분시킨 2단계 CSTR/TBF 시스템을 개발하였다. B. ceapcia G4를 분해 미생물로 사용하였으며, 탄소원인 phenol과 공기 및 배지가연속적으로 공급되는 CSTR에서 효소 및 세포 재활성를 도모하고, TBF에서는 폐가스에 포함된 기사의 TCE를 공기와 함께 공급하여 B. cepcia G4 미생물막에의한 TCE가 분해되도록 구성하였다. 2단계 CSTR/TBF 시스템은 유입 TCE 농도가 15pp $m_{v}$까지도 100% 수준의 처리 효율을 보여주었으며, 고농도의 TCE를 안정적으로 장기간 처리할 수 있었다.다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.804-811
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• 2001

In many Gram-negative bacteria, autoinducers, such as N-acyl-L-homoserine lactone(acyl-HSL) and its derivative molecules, mediate the cell-density-dependnet expression of certain operons. The current study identified the autoinducers produced by Burkholderia cepacia G4, a trichloroethylene-degrading lagoon isolate, using TLC bioassays with Agrobacterium tumefaciens NT1(pDCI141E33) and Chromobacterium violaceum CVO26, and a GC-MS analysis. The ${R_f}\;and\;{R_t}$ values and mass spectra were compared with those of synthetic compounds. Based on the analyses, it was confirmed that G4 produces N-hexanoyl (C6)-, N-octanoyl (C8)-, N-decanoyl (C10)-, N-dodecanoyl (C12)-HSL, and an unknown active species. The integration of the GC peak areas exhibited a ratio of C8-HSL:C10-HSL:C12-HSL at 3:17:1 with C6-HSL and C10-HSL production at trace and micromolar levels, respectively, in the culture supernatants. Nutants partially defective in producing acyl-HSLs were also partially defective in the biosynthesis of an antibiotic substance. These results indicate that the autoinducer-dependent gene regulation in G4 is dissimilar to the clinical B. cepacia strains isolated from patients with cystic fibrosis.

• Archives of Pharmacal Research
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• 제27권5호
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• pp.570-575
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• 2004

2-Hydroxymuconic semialdehyde (2-HMS) dehydrogenase catalyzes the conversion of 2-HMS to 4-oxalocrotonate, which is a step in the meta cleavage pathway of aromatic hydrocarbons in bacteria. A tomC gene that encodes 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol, or benzene, was overexpressed into E. coli HB 101, and its gene product was characterized in this study. 2-HMS dehydrogenase from B. cepacia G4 has a high catalytic efficiency in terms of V$_{max}$K$_{max}$ towards 2-hydroxy-5-methyl-muconic semialdehyde followed by 2-HMS but has a very low efficiency for 5-chloro-2-hydroxymuconic semialdehyde. However, the enzyme did not utilize 2-hydroxy-6-oxo-hepta 2,4-dienoic acid and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid as substrates. The molecular weight of 2-HMS dehydrogenase from B. cepacia G4 was predicted to be 52 kDa containing 485 amino acid residues from the nucleotide sequence of the tomC gene, and it exhibited the highest identity of 78% with the amino acid sequence of 2-HMS dehydrogenase that is encoded in the aphC gene of Comamonas testosteroni TA441. 2-HMS dehydrogenase from B. cepacia G4 showed a significant phylogenetic relationship not only with other 2-HMS dehydrogenases, but also with different dehydrogenases from evolutionarily distant organisms.sms.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.218.2-219
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• 2003

2-Hydroxymuconic semialdehyde dehydrogenase catalyzes the conversion of 2-hydroxymuconic semialdehyde (HMS) to an enol form of 4-oxalocrotonate which is a step in the catechol-meta cleavage pathway. A tomC gene encoding 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol or tricholoro ethylene, is identified in between catechol 2,3-dioxygenase gene and HMS hydrolase gene, its sequence is analysed and the enzyme is characterised. (omitted)

• KSBB Journal
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• 제16권5호
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• pp.466-473
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• 2001

분리 규주는 운동성을 갖는 그람 음성 간균으로서 catalase 양성 반응을 보였으며, citrate, mannitol, sucorse, frutose, trehaslos 등을 이용하였다. Biolog test 결과, 분리 균주는 burkholderia(Pseudomonas) cepacia 이 것으로 동정되었으며, 85.5%의 유사성을 나타내었다. 분리 균주의 최적온도와 pH는 각각 3$0^{\circ}C$, 7.0이었으며, pH 4.0-8.0의 넓은 범위에서 성장이 가능하였다. 그리고 분리 균주의 유기물의 영향을 알아 본 결과 thiosulfate 배지에 yeast extract를 유일한 탄소원으로 혼합하여 첨가하면 thiosulfate 와 yeast extract 중 하나가 성장 제한 인자 중 하나의 농도를 증가시키면 세포 성장에 따른 균체량이 증가하였다. 본 실험에서는 유기물로는 glucose 보다 yeast extract를 첨가했을 때가 증식 속도와 황산화 속도가 높게 나타났다. Thiosulfate 농도에 따른 분리균주의 황산화 속도와 최대 황산화 속도는 각각 0.56.h$^{-1}$과 0.18 g-S/L.h로 나타났다. 결과는 환산화 속도는 기질 농도가 증가함에 따라 완만하게 증가하여 0.12M에서 0.2 g-S.L.에 도달하였으나 0.16M에서는 급격하게 감소하였다. 분리균의 황하 수소 제거능을 조사하기 위하여 각 flow rate 에 따른 최대 제거 속도는 Vm과 K CIn/R과 CIn의 그래프를 이용하여 Hanes-Wolf 식을 통하여 구했다. 즉, 그래프 기울기와 Y절편으로부터 계산한 flow rate 12 L/h에서의 황화 수소에 대한 Vm과 Ks는 각각 6.25 g-S.㎤.h$^{-1}$과 22.88ppm이다 높은 flow rate에서의 제거 효율과 제거 용량은 역 경향이 나타났다. 이것은 mass transfer 효과 즉, 확산 제한에 기인한 것이지 미생물의 효소 활성의 제한에 의한 것은 아니다 (19), 따라서 biofilter 내로 고농도의 황화 수소가 유입 될 때 황화 수소 가스의 flow rate를 줄이든지 또는 적절한 제거 용량에 도달하기 위해 충진물의 부피를 증가 할 필요가 있다.

• KSBB Journal
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• 제15권4호
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• pp.393-397
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• 2000

The cometaboic biodegradation conditionso f trichloroethylene(TCE) by Burkholderia cepacia G4 were optimized using response surface analysis. The experimental sets of phenol concentration temperature and pH were designed using central composite experimental design. The optimal conditions of phenol concentration temperature and pH were determined to be 0.91 ppm 21.5$^{\circ}C$ and 7.65 respectively by the Ridge analysis of the contour plot for TCE biodegradation rates. The TCE biodegradation rate could be enhanced up to 2.43 nmol.mg protein$.$min by response surface methodology.