• Journal of Microbiology
• /
• 제41권4호
• /
• pp.349-352
• /
• 2003

Recent studies have shown that some of the PCB (polychlorinated biphenyl)-degraders are able to effectively degrade PCB in the presence of monoterpenes, which act as inducers for the degradation pathway. Rhodococcus sp. T104, an effective PCB degrader, has been shown to induce the degradation pathway by utilizing limonenes, cymenes, carvones, and pinenes as sole carbon sources which can be found in the natural environment. Moreover, the strain T104 proved to possess three separate oxidation pathways of limonene, biphenyl, and phenol. Of these three, the limonene can also induce the biphenyl degradation pathway. In this work, we report the presence of three distinct genes for aromatic oxygenase, which are putatively involved in the degradation of aromatic substrates including biphenyl, limonene, and phenol, through PCR amplification and denaturing gradient gel electrophoresis (DGGE). The genes were differentially expressed and well induced by limonene, cymene, and plant extract A compared to biphenyl and/or glucose. This indicates that substrate specificity must be taken into account when biodegradation of the target compounds are facilitated by the plant natural substrates.

• Journal of Microbiology
• /
• 제40권1호
• /
• pp.86-89
• /
• 2002

Rhodococcus sp. strain T104, which is able to grow on either biphenyl or limonene, was found to utilize phenol as sole carbon and energy sources. Furthermore, T104 was positively identified to possess three separate pathways for the degradation of limonene, phenol, and biphenyl. The fact that biphenyl and limonene induced almost the same amount of catechol 1,2-dioxygenase activity indicates that limonene can induce both upper and lower pathways for biphenyl degradation by T104.

• 한국환경과학회지
• /
• 제19권11호
• /
• pp.1391-1396
• /
• 2010

Three biphenyl-degrading microorganisms were isolated from polluted soil samples in Sasang-gu, Busan. Among them, isolate DS-94 showing the strong degrading activity was selected. The morphological, physiological, and biochemical characteristics of DS-94 were investigated by API 20NE and other tests. This bacterium was identified as the genus Pseudomonas by 16S rDNA sequencing and designated as Pseudomonas sp. DS-94. The optimum temperature and pH for the growth of Pseudomonas sp. DS-94 were $25^{\circ}C$ and pH 7.0, respectively. This isolate could utilize biphenyl as sole source of carbon and energy. Biphenyl-degrading efficiency of this isolate was measured by HPLC analysis. As a result of biological biphenyl-degradation at high biphenyl concentration (500 mg/L), biphenyl-removal efficiency by this isolate was 73.5% for 7 days.

• Journal of Microbiology
• /
• 제42권2호
• /
• pp.160-162
• /
• 2004

Rhodococcus sp. strain T104 is able to utilize both limonene and biphenyl as growth substrates. Fur-thermore, T104 possesses separate pathways for the degradation of limonene and biphenyl. Previously, we found that a gene(s) involved in limonene degradation was also related to indigo-producing ability. To further corroborate this observation, we have cloned and sequenced a 8,842-bp genomic DNA region with four open reading frames, including one for indole oxygenase, which converts indole to indigo (a blue pigment). The reverse transcription PCR data demonstrated that the identified indole oxygenase gene is specifically induced by limonene, thereby implicating this gene in the degradation of limonene by T104.

• 한국지하수토양환경학회:학술대회논문집
• /
• 한국지하수토양환경학회 2005년도 총회 및 춘계학술발표회
• /
• pp.28-30
• /
• 2005

Because of high population diversity in soil microbial communities, it is difficult to accurately assess the capability of biodegradation of toxicant by microbes in soil and sediment. Identifying biodegradative microorganisms is an important step in designing and analyzing soil bioremediation. To remove non-important noise information, it is necessary to selectively enrich genomes of biodegradative microorganisms fromnon-biodegradative populations. For this purpose, a stable isotope probing (SIP) technique was applied in selectively harvesting the genomes of biphenyl-utilizing bacteria from soil microbial communities. Since many biphenyl-using microorganisms are responsible for aerobic PCB degradation In soil and sediments, biphenyl-utilizing bacteria were chosen as the target organisms. In soil microcosms, 13C-biphenyl was added as a selective carbon source for biphenyl users, According to $13C-CO_2$ analysis by GC-MS, 13C-biphenyl mineralization was detected after a 7-day of incubation. The heavy portion of DNA(13C-DNA) was separated from the light portion of DNA (12C-DNA) using equilibrium density gradient ultracentrifuge. Bacterial community structure in the 13C-DNAsample was analyzed by t-RFLP (terminal restriction fragment length polymorphism) method. The t-RFLP result demonstates that the use of SIP efficiently and selectively enriched the genomes of biphenyl degrading bacteria from non-degradative microbes. Furthermore, the bacterial diversity of biphenyl degrading populations was small enough for environmental genomes tools (metagenomics and DNA microarrays) to be used to detect functional (biphenyl degradation) genes from soil microbial communities, which may provide a significant progress in assessing microbial capability of PCB bioremediation in soil and groundwater.

• Journal of Microbiology
• /
• 제35권2호
• /
• pp.141-148
• /
• 1997

Alcaligenes xylosoxydans strain SMN3 capable of utilizing biphenyl grew not only on phenol, and benzoate, but also on salicylate. Catabolisms of biphenyl and salicylate appear to be interrelated since benzoate is a common metabolic intermediate of these compounds. Enzyme levels in the excatechol 2. 3-dioxygenas which is meta-cleavage enzyme of catechol, but did not induce catechol 1, 2-dioxygenase. All the oxidative enzymes of biphenyl and 2, 3,-dihydroxybiphenyl (23DHBP) were induced when the cells were grown on biphenyl and salicylate, respectively. Biphenyl and salicylate could be a good inducer in the oxidation of biphenyl and 2, 3-dihydroxybiphenyl. The two enzymes for the degradation of biphenyl and salicylate were induced after growth on either biphenyl or salicylate, suggesting the presence of a common regulatory element. However, benzoate could not induce the enzymes responsible for the oxidation of these compounds. Biphenyl and salicylate were good inducers for indigo formation due to the activity of biphenyl dioxygenase. These results suggested that indole oxidation is a property of bacterial dioxygenase that form cis-dihydrodiols from aromatic hydrocarbon including biphenyl.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
• /
• pp.489-492
• /
• 2000

The eventual goal of this study is to elucidate roles of plant terpenoids (e.g., cymene, limonene and others) as natural substrates in the cometabolic biodegradation of PCBs and to develop an effective PCB bioremediation technology. The aim of this study was to examine how plant terpenoids, as natural substrates or inducers would affect the biodegradation of PCB congeners. Various PCB degraders that could grow on biphenyl and several terpenoids were tested for their PCB degradation capabilities. The PCB congener degradation activities were first monitored through resting cell assay technique that could detect degradation products of the substrate. The congener removal was also confirmed by concommitant GC analysis. The PCB degraders, Pseudononas sp. P166 and Caynebacterium sp. T104 were found to grow on both biphenyl and terpenoids ((S)-(-) limonene, p-cymene and ${\alpha}-terpinene$) whereas Arthrobacter B1B could not grow on the terpenoids as a sole carbon source. The strain B1B grown on biphenyl showed a good degradation activity for 4,4'-dichlorobiphenyl (DCBp) while strains P166 and T104 gave about 25% of B1B activity. Induction of degradation by cymene, limonene and terpine was hardly detected by the resting cell assay technique. This appeared to be due to relatively lower induction effect of these terpenoids compared with biphenyl. However, a subsequent GC analysis showed that the congener could be removed up to 30% by the resting cells of T104 grown on the terpenoids. This indicates that terpenoids, widely distributed in nature, could be utilized as both growth and/or inducer substrate for PCB biodegradation.

• 한국미생물·생명공학회지
• /
• 제34권2호
• /
• pp.174-179
• /
• 2006

PCBs(polychlorinated biphenyl)는 난분해성 물질로써, 환경호르몬으로 분류된 유독한 화합물이다. 이런 유독한 화합물인 PCBs 화합물이 오염된 토양 및 수계를 회복하기 위해 PCBs의 모체인 biphenyl을 효과적으로 분해하는 미생물을 토양으로부터 분리 선별하여 S. yanoikuyae BK10 (AF406817)와 같이 분해능이 우수한 균주를 분리하였다. 분리된 S. yanoikuyae BK10의 특성을 조사하기 위하여 자연계의 토양 조건인 pH 5.0$\sim$8.0에서 99%이상의 높은 biphenyl 분해효율을 보였다. 또한, 온도를 달리하여 실험 한 결과, 10$\sim$50$^{\circ}C$의 범위에서 모두 70%이상의 높은 분해효율을 보여줌으로써 실제 biphenyl/PCBs로_오염된 토양에서 온도의 영향을 덜 받고 biphenyl을 효과적으로 분해 할 수 있을 것으로 생각된다. S. yanoikuyae BK10는 biphenyl이 500 $\mu$g/ml으로 처리된 mineral salt 배지에서 48시간동안 99% 이상의 biphenyl을 분해하는 높은 분해활성을 보이며, biphenyl을 mineralization 시키는 것으로 판단된다. 또한 biphenyl 분해효소 유도 실험결과는 기질을 biphenyl로 사용하여 증식한 균체가 다른 기질을 사용해서 증식한 균체보다 약 2배가량 biphenyl을 빨리 분해시켰다. 그렇지만, cell-mass를 많이 얻을 수 있는 당을 탄소원으로 사용하여 배양하였을 때에도 단시간 내에 biphenyl분해 효소를 분비하여 biphenyl을 분해하는 것으로 보아, S. yanoikuyae BK10는 실제 biphenyl/PCBs에 오염된 토양 적용 할 경우 안정적으로 균주의 제공이 가능하다고 판단된다. 이상의 결과를 토대로, 토양에서부터 분리한 S. yanoikuyae BK10는 자연계에서 유해화합물인 biphenyl/PCBs을 효과적으로 분해 할 수 있다고 생각되며, 분리균주인 S. yanoikuyae BK10의 분자 생물학적 특성을 조사하여 biphenyl과 PCBs를 분해하는 유전자 탐색에 유용한 정보를 얻을 수 있을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
• /
• 제26권11호
• /
• pp.1943-1950
• /
• 2016

Polycyclic aromatic hydrocarbons (PAHs) are commonly present xenobiotics in natural and contaminated soils. We studied three (phenanthrene, naphthalene, and biphenyl) xenobiotics, catabolism, and associated proteins in Sphingobium chungbukense DJ77 by two-dimensional gel electrophoresis (2-DE) analysis. Comparative analysis of the growth-dependent 2-DE results revealed that the intensity of 10 protein spots changed identically upon exposure to the three xenobiotics. Among the upregulated proteins, five protein spots, which were putative dehydrogenase, dioxygenase, and hydrolase and involved in the catabolic pathway of xenobiotic degradation, were induced. Identification of these major multifunctional proteins allowed us to map the multiple catabolic pathway for phenanthrene, naphthalene, and biphenyl degradation. A part of the initial diverse catabolism was converged into the catechol degradation branch. Detection of intermediates from 2,3-dihydroxy-biphenyl degradation to pyruvate and acetyl-CoA production by LC/MS analysis showed that ring-cleavage products of PAHs entered the tricarboxylic acid cycle, and were mineralized in S. chungbukense DJ77. These results suggest that S. chungbukense DJ77 completely degrades a broad range of PAHs via a multiple catabolic pathway.

• Archives of Pharmacal Research
• /
• 제24권2호
• /
• pp.159-163
• /
• 2001

The accelerated stability of dimethoxy biphenyl monocarboxylate.HCl (DDB-S) was investigated in 6 mg/mL water solution in the pH ranging 2-10 and the temperature of $45-85^{\circ}C$. The observed rate of degradation followed first-order kinetics. The energy of activation for DDB-S degradation was calculated to be 14.1 and 16.5 $Kcal/mole$ at pH 5 and in distilled watery respectively. The degradation rate constant ($K_{25^{\circ}C}$) obtained by trending line analysis of Arrhenius plots for DDB-S was $5.3{\times}10^{-6}h^{-1}$. The times to degrade 10% ($t_{10}$) and 50% $t_{500}$) at $K_{25^{\circ}C}$ were 829 and 5,416 days, respectively. DDB-S exhibited the fastest degradation at pH 10 and the slowest rate at pH 5. In addition, at $K_{65^{\circ}C}$, degradation rate constants of DDB-S were 0.066, 0.059, 5.460, 32.171, and $1.4{\times}10^{-6}h^{-1}$ at pH 2, 5, 8, 10 and in distilled water, respectively. These observations indicated that the rate-pH profile of DDB-S showed general acid-base catalysis reaction in the range of pH 2-10.