• Journal of Applied Biological Chemistry
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• 제63권2호
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• pp.125-129
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• 2020

Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

• Journal of Microbiology and Biotechnology
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• 제30권7호
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• pp.1005-1012
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• 2020

Acid mine drainage (AMD) has been a serious environmental issue that threatens soil and aquatic ecosystems. In this study, an acid-tolerant sulfate-reducing bacterium, strain S4, was isolated from the mud of an AMD storage pond in Vietnam via enrichment in anoxic mineral medium at pH 5. Comparative analyses of sequences of the 16S rRNA gene and dsrB gene involved in sulfate reduction revealed that the isolate belonged to the genus Desulfovibrio, and is most closely related to Desulfovibrio oxamicus (with 99% homology in 16S rDNA sequence and 98% homology in dsrB gene sequence). Denaturing gradient gel electrophoresis (DGGE) analyses of dsrB gene showed that strain S4 represented one of the two most abundant groups developed in the enrichment culture. Notably, strain S4 was capable of reducing sulfate in low pH environments (from 2 and above), and resistance to extremely high concentration of heavy metals (Fe 3,000 mg/l, Zn 100 mg/l, Cu 100 mg/l). In a batch incubation experiment in synthetic AMD with pH 3.5, strain S4 showed strong effects in facilitating growth of a neutrophilic, metal sensitive Desulfovibrio sp. strain SR4H, which was not capable of growing alone in such an environment. Thus, it is postulated that under extreme conditions such as an AMD environment, acid- and metal-tolerant sulfate-reducing bacteria (SRB)-like strain S4 would facilitate the growth of other widely distributed SRB by starting to reduce sulfate at low pH, thus increasing pH and lowering the metal concentration in the environment. Owing to such unique physiological characteristics, strain S4 shows great potential for application in sustainable remediation of AMD.

• 한국환경농학회지
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• 제16권4호
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• pp.311-319
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• 1997

광산폐수, 산업폐수등으로 부터 Cd, Pb, Zn 및 Cu등의 중금속에 강한 내성을 지니고 있을 뿐만 아니라 균체내 중금속 축적능력이 우수한 중금속 내성 미생물 균주 Pseudomonas putida(Cd), Pseudomonas aeruginosa(Pb), Pseudomonas chlororaphis(Zn) 및 Pseudomonas stutzeri(Cu)를 각각 분리하여, 여러가지 물리화학적인 방법으로 세포를 전처리하여 세포구성성분을 인위적으로 조절한 후 세포내 중금속이온의 흡수 거동 및 조단백질 함량과 중금속 축적관계를 조사한 결과는 다음과 같다. 세포를 알카리로 전처리하였을 경우 세포내 중금속 축적은 매우 감소되었으며, 메탄올과 클로포름으로 전처리하였을 경우에는 중금속 축적에 큰 영향을 미치지 않았으나, 메탄올과 클로로포름으로 전처리한 후 다시 알카리로 재차 처리하였을 경우에는 중금속 축적이 매우 감소되었다. 전처리된 세포내 중금속 축적은 용출되지 않고 남아있는 조단백질 함량이 감소됨에 따라 더 얼마나 크게 감소되었으므로 세포 구성물질중 단백질이 중금속 축적에 중요한 역할을 하는 물질인 것으로 판단되었다.

• Journal of Microbiology and Biotechnology
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• 제28권7호
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• pp.1156-1167
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• 2018

The aim of this study was to isolate and characterize lead (Pb)-solubilizing bacteria from heavy metal-contaminated mine soils and to evaluate their inoculation effects on the growth and Pb absorption of Brassica juncea. The isolates were also evaluated for their plant growth-promoting characteristics as well as heavy metal and salt tolerance. A total of 171 Pb-tolerant isolates were identified, of which only 15 bacterial strains were able to produce clear haloes in solid medium containing PbO or $PbCO_3$, indicating Pb solubilization. All of these 15 strains were also able to dissolve the Pb minerals in a liquid medium, which was accompanied by significant decreases in pH values of the medium. Based on 16S rRNA gene sequence analysis, the Pb-solubilizing strains belonged to genera Bacillus, Paenibacillus, Brevibacterium, and Staphylococcus. A majority of the Pb-solubilizing strains were able to produce indole acetic acid and siderophores to different extents. Two of the Pb-solubilizing isolates were able to solubilize inorganic phosphate as well. Some of the strains displayed tolerance to different heavy metals and to salt stress and were able to grow in a wide pH range. Inoculation with two selected Pb-solubilizing and plant growth-promoting strains, (i.e., Brevibacterium frigoritolerans YSP40 and Bacillus paralicheniformis YSP151) and their consortium enhanced the growth and Pb uptake of B. juncea plants grown in a metal-contaminated soil. The bacterial strains isolated in this study are promising candidates to develop novel microbe-assisted phytoremediation strategies for metal-contaminated soils.

• 한국지하수토양환경학회지:지하수토양환경
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• 제20권4호
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• pp.113-121
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• 2015

The contaminated soil at abandoned smelter areas present challenge for remediation, as the degraded materials are typically deficient in nutrients, and rich in toxic heavy metals and metalloids. Bioremediation technique is to isolate new strains of microorganisms and develop successful protocols for reducing metal toxicity with heavy metal tolerant species. The present study collected metal contaminated soil and characterized for pH and EC values, and heavy metal contents. The pH value was 5.80, representing slightly acidic soil, and EC value was 13.47 mS/m. ICP-AES analytical results showed that the collected soil samples were highly contaminated with various heavy metals and metalloids such as lead (183.0 mg/kg), copper (98.6 mg/kg), zinc (91.6 mg/kg), and arsenic (48.1 mg/kg), respectively. In this study, a bacterial strain, Bacillus cereus KM-15, capable of adsorbing the heavy metals was isolated from the contaminated soils by selective enrichment and characterized to apply for the bioremediation. The effects of heavy metal on the growth of the Bacillus cereus KM-15 was determined in liquid cultures. The results showed that 100 mg/L arsenic, lead, and zinc did not affect the growth of KM-15, while the bacterial growth was strongly inhibited by copper at the same concentration. Further, the ability of the bacteria to adsorb heavy metals was evaluated.

• 한국토양비료학회지
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• 제47권3호
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• pp.213-216
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• 2014

Arsenic is a known hazardous metalloid not only to the animals but also to plants. With high concentrations, it can impede normal plant growth and cause even death of plants at extremely high levels. A known plant response to stress conditions such as toxic levels of metal (loids) is the production of stress ethylene, causing inhibitory effect on root growth in plants. When the effect of various arsenic concentrations was tested to maize plant, the stress ethylene emission proportionately increased with increasing concentration of As(V). The inoculation of two arsenic tolerant bacteria; Pseudomonas grimonti JS126 and Pseudomonas taiwanensis JS238 having respective high and low 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity reduced stress ethylene emission by 59% and 30% in maize grown in arsenic polluted soils. The result suggested the possible use of Pseudomonas grimonti JS126 for phytoremediation of arsenic polluted soils.

• 생명과학회지
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• 제8권3호
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• pp.339-347
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• 1998

As a part of a study on the development of microbiological methods for petroleum exploration, the distribution of the avarafe survival rate for heterotrophs to various heavy metal ions and hydrocarbons were surveyed and compared by the use of plate count method. In consequence of the avarage survival rate to heavy metal ions(2 hours treatment) and hydrocarbons(1 hour treatment) for heterotrophs isolatinf from soil samples(50cm depth) which located in Doum mountain(A, B and D site) and Aedowon(C site) at Pohang area, the survival rate of heterotrophs for nickel(600ppm), cobalt(500ppm), cadmiun(100ppm), mercury(20ppm), zinc(400 ppm) and lead(500ppm) were 73.7%, 82.6%, 76.8%, 9.5%, 77.8% and 73.6% at A site and 67.9%, 82.5%, 86.0%, 5.8%, 82.5% and 91.7% at B site, 87.8%, 79.8%, 87.5%, 7.0%, 84.2% AND 47.7% AT c SITE, AND 71.8%, 76%, 85.9%, 1,2%, 79.6% AND 88.3% AT D site, respectively. Also the survival rate of heterotrophs from A,B,C and D site to pentane and hexane(each concentration is 20%) were 26.7% and 42.5%, 11.8% and 8.1%, 44.3% and 36.2%, and 12% and 3.5%, respectively. therefore, heterotrophs from B and D site that alternated gravelstone, muddy sandstone and sandstone were higher survival rate to the heavy metal ions than heterotrophs from A site which mainly composed gravelstone. Also, heterotrophs from C site which mainly composed muddy sandstone and once produced natural gas were showed relatively higher survival rate to the heavy metal ions and hydrocarbons than the other sites. Consequently, we confirmed that the distributions of tolerant heterotrophs to heavy metal ions and hydrocarbons were differ from the lithological compositon.

• 한국토양비료학회지
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• 제45권5호
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• pp.817-821
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• 2012

Soil samples collected from abounded mines of Boryeong area in South Korea were used in isolating bacterial strains and their capacity to solubilize inorganic phosphates and heavy metal tolerance were assessed in vitro. Three different inorganic phosphate sources (Ca phosphate, Fe phosphate, and Al phosphate) and four different heavy metals (Co, Cd, Pb and Zn) each with three concentrations ($100{\mu}g\;mL^{-1}$, $200{\mu}g\;mL^{-1}$, and $400{\mu}g\;mL^{-1}$) were used. The bacterial isolates PSB-1, PSB-2, PSB-3, and PSB-4 solubilized significantly higher amount of Ca phosphate during the first five days of incubation though subsequent drop in soluble phosphorus level in the medium was observed at the later stage (after 5 days) of the incubation. Solubilization of Ca phosphate and Fe phosphate was concomitant with the acidification of the culture medium compared to the control where it remained constant. Isolated strains could solubilize Fe phosphate to certain extent ($25-45{\mu}g\;mL^{-1}$) though solubilization of Al phosphate was found negligible. All the isolates were tolerant to heavy metals (Cd, Pb, and Zn) up to the concentration of $400{\mu}g\;mL^{-1}$ except PSB-1 and PSB-8, which were shown to be vulnerable to Co even at $100{\mu}g\;mL^{-1}$. Heavy metal tolerant strains should be further evaluated for plant growth promoting activities also under field conditions in order to assess their agricultural and environmental significance.

• Ocean and Polar Research
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• 제25권1호
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• pp.21-30
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• 2003

Seasonal fluctuations of physico-chemical and biological aspects of the environment were studied in Vladivostok harbour (Golden Horn Bay, the East Sea/Sea of Japan). The benthic community structure was described with a focus on size-spectra (bacteria, meio- and macrofauna) related with the chemical environment and chemical fluxes in sediment and to reveal their possible ecological role in the process of bioremediation of the environment. Samples from two sites with different concentrations of heavy metals (Fe, Zn, Cu, Pb, Mn, Cr, Ni Cd, Co) and petroleum hydrocarbon were assessed by a number of methods. These included plate counts of culturable bacteria, observation through a scanning electron (SEM) and transmission electron microscope (TEM). These approaches were complemented with microscopic assessments of the diversity of the benthic community. The specific communities had a limited number of species, tolerant to abnormally high levels of toxic compounds. The dominant species were presented by several sho.1-lived small polychaetes (Capitella capitata) and nematodes (Oncholaimium ramosum). The highest population density was recorded in microbenthos, in various diatoms, various physiological groups of bacteria which participate in biomineralization: marine heterotrophic bacteria, which oxidized oil, black oil in addition to groups resistant to heavy metals. They have the entire set of mechanisms for neutralizing the negative effect of those compounds, forming the detrital food web and biogeochemical circulation of material in sediments, which results in the biological self-recycling of sea basins. Macro- and meiobenthic organisms were more sensitive to a greater extent of $H_2S$ and petroleum hydrocarbons than to metal content, but the within-site rankings were the same as those achieved for microbiological analyses.

• 대한환경공학회지
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• 제28권11호
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• pp.1222-1227
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• 2006

중금속으로 오염된 폐 광산에서 중금속(카드뮴, 구리)에 내성을 가지는 총 9개의 균주를 순수 분리한 후, 스크린 테스트를 거쳐 고효율의 중금속 제거 균주 JH1을 선별하였다. JH1 균주는 형태학적, 생리 생화학적 특징, 지방산 분석 그리고 16S rDNA 분석결과 Ralstonia eutropha JH1으로 동정되었다. 카드뮴으로 오염된 토양을 구연산(10 mM, pH 6.0)으로 세척 한 후, 그 세척수(카드뮴 110 mg/L)에서 균 농도(0.5, 1.0, 2.0, 4.0 g/L)에 따른 카드뮴 제거율을 확인한 결과, 5일 동안 각각 49.9, 84.4, 89.7, 89.9% 제거되었다. 또한 카드뮴(110 mg/L)과 구리(100 mg/L)를 함유하고 있는 구연산-세척수에서 Ralstonia eutropha JH1을 1.0 g/L로 접종했을 때, 6일 동안 카드뮴과 구리가 100% 제거 되었다. 저농도(10, 30, 60 mg/L)의 카드뮴과 구리를 각각 함유한 세척수 내에 Ralstonia eutropha JH1을 1.0 g/L로 접종하고 시간에 따른 제거율을 확인한 결과, 카드뮴과 구리 모두 10, 30, 60 mg/L에서 각각 12, 18, 48시간에서 100%제거되었다.