• 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.

• 한국토양비료학회지
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• 제37권6호
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• pp.371-377
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• 2004

산림토양에서의 황산이온 흡착은 식물의 흡수, 양이온 이동 및 산중화능 등 산림생태계의 황 동태에 중요한 영향을 미친다. 따라서 본 연구는 산림토양 중의황산이온 분포와 그와 관련된 황산이온 흡착이 어떠한 토양특성에 의해 영향을 받는지를 밝히기 위해 수행되었다. 이를 위해 소나무와 신갈나무 임분으로 조성된 4개 산림지역을 대상으로 표토층과 심토층으로 구분하여 황산이온 흡착지표로서의 추출성 황산이온함량 및 황산이온 흡착능을 정량 한 후 관련 토양 물리화학적 특성인 pH. 양이온치환용량, 점토함량 및 Ai과 Fe 산화물의 종류별 함량과의 상관성을 분석하였다. 토양 중 추출성 황산이온 함량 및 황산이온 흡착능은 각 조사지별로 큰 차이를 보였으며 전체적으로 유기물 함량이 적고 Al 또는 Fe 산화물 함량은 많은 심토층이 표토층에 비하여 황산이온 흡착량이 많았다. 황산이온 흡착량이 많았던 심토층의$Al_d$와 $Fe_d$ 산화물 평균 함량은 각각 8.49와 $12.45g\;kg^{-1}$로 대체로 낮은 수준이었다. 황산이온 흡착지표들은 토양 pH, 양이온치환용량 및 점토함량과 대체로 유의적인 정의 상관관계를 보였다. 반면에 유기탄소 함량과는 부의 상관성을 보여 산림토양 중에서 유기물이 황산이온 흡착에 대해 경합물질로 작용함을 알 수 있었다. Ai과 Fe 산화물 중에서는 $Al_d,\;Al_o,\;Al_p,\;Al_a$ 및 $Fe_a$가 황산이온 흡착지표들과 유의성 있는 상관성을 보였으나, $Al_c$와 $Fe_c$는 황산이온 흡착지표들과 상관성이 없었다. 황산이온 흡착지표인자들과 토양 pH, 양이온치환용량 및 비결정형 Ai 함량간의 정의 상관관계 결과는 토양산성화로 인해 산림생태계에서 황 동태에 기여하는 산림토양의 황산이온 흡착 능력이 저하될 수 있음을 반영한다.

• 한국지반공학회논문집
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• 제21권5호
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• pp.83-92
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• 2005

산성암반배수가 발생되는 지역을 중심으로 사면구조물의 피해현황과 환경오염 실태를 파악한 결과를 하면 다음과 같다. 첫째, 산성암반배수가 발생하는 절취사면은 대부분 숏크리트공법으로 처리하고 있으며, 산성암반배수가 사면구조물의 안정성에 악영향을 미치고 있다. 둘째, 숏크리트는 산성암반배수에 의해서 현저하게 열화되어 가고 있으며, 용수발생량이 많은 곳은 배면공동이 발견되는 경우도 있다. 셋째, 절취사면에서 산성암반배수가 발생되는 곳은 국부적으로 불안정하고, 이를 보강하기 위하여 앵커와 록볼트를 사용하고 있으나 강산성암반배수에 의해 부식될 우려가 크다. 넷째, 식생공의 경우 산성암반배수가 잔디 씨앗의 발아 및 성장에 악영향을 미치고 있어 일부 지역에서는 생석회를 사용하여 산성암반배수를 중화시키고 있으나 근본적인 대책은 되지 못하고 있다. 다섯째, 황철석의 풍화로 발생한 산에 의해 배수가 낮은 pH를 유지하면서 중금속들을 용출시켜 지속적으로 배출하여 부근 토양, 지하수와 하천수를 오염시킬 개연성이 매우 크다. 산성암반배수의 발생 가능성을 암석유형별로 평가한 결과 편마암, 화강암의 경우 산성배수 발생 가능성이 낮은 군으루 열수변질을 받근 화산암, 응회암, 탄질셰일, 금속광산 폐석시료는 산성배수 발생 가능성이 높은 군으로 분류되었다. 따라서 토목건설공사과정에서 빈번히 발생될 것으로 예상되는 사면절취에 의한 암반산성배수에 대비하여 발생개연성이 있는 지역에 대한 지반정보의 확보 및 대책기술의 개발이 요구되고있다.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.534-544
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• 2014

The effects of microbial iron reduction and oxidation on the immobilization and mobilization of copper were investigated in a high concentration of sulfate with synthesized Fe(III) minerals and red earth soils rich in amorphous Fe (hydr)oxides. Batch microcosm experiments showed that red earth soil inoculated with subsurface sediments had a faster Fe(III) bioreduction rate than pure amorphous Fe(III) minerals and resulted in quicker immobilization of Cu in the aqueous fraction. Coinciding with the decrease of aqueous Cu, $SO_4{^{2-}}$ in the inoculated red earth soil decreased acutely after incubation. The shift in the microbial community composite in the inoculated soil was analyzed through denaturing gradient gel electrophoresis. Results revealed the potential cooperative effect of microbial Fe(III) reduction and sulfate reduction on copper immobilization. After exposure to air for 144 h, more than 50% of the immobilized Cu was remobilized from the anaerobic matrices; aqueous sulfate increased significantly. Sequential extraction analysis demonstrated that the organic matter/sulfide-bound Cu increased by 52% after anaerobic incubation relative to the abiotic treatment but decreased by 32% after oxidation, indicating the generation and oxidation of Cu-sulfide coprecipitates in the inoculated red earth soil. These findings suggest that the immobilization of copper could be enhanced by mediating microbial Fe(III) reduction with sulfate reduction under anaerobic conditions. The findings have an important implication for bioremediation in Cu-contaminated and Fe-rich soils, especially in acid-mine-drainage-affected sites.

• 한국지하수토양환경학회지:지하수토양환경
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• 제18권7호
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• pp.81-89
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• 2013

Concerns have been raised over the impact of nano materials on soil and groundwater environment with the increasing attention to the potential applications of carbon nano materials in various fields. Particularly, carbon nano materials introduced into water environment readily make complexes with humic acid (HA) due to their hydrophobic nature, so there have been increasing numbers of studies on the interaction between HA and carbon nano materials. In this study, we investigated the solubility of HA and multiwalled carbon nanotubes (MWCNT) in three different surfactant solutions of sodium dodecyl sulfate (SDS), Brij 30 and Triton X-100, and evaluated whether the HA can be effectively desorbed from the surface of MWCNT by surfactant. The objective of this study was to determine the optimal adsorption condition for HA to MWCNT. Futhermore, sodium dodecyl sulfate (SDS), Brij 30, Triton X-100 were used to elucidate the effect of desorption and separation on adsorbed HA on MWCNT. As a result, HA solution with 12.7 mg of total organic carbon (TOC) and 5 mg of MWCNT showed the highest adsorption capacity at pH 3 reacted for 72 hrs. Weight solubilizing ratio (WSR) of surfactants on HA and MWCNT was calculated. HA had approximately 2 times lower adsorption capacity for the applied three surfactants compared to those of MWCNT, implying that the desorption of HA may occur from the HA/MWCNT complex. According to the results of adsorption isotherm and weight solubilizing ratio (WSR), the most effective surfactants was the SDS 1% soluiton, showing 53.63% desorption of HA at pH 3.

• 한국농림기상학회지
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• 제2권4호
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• pp.190-197
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• 2000

산성비와 같은 요인에 의해 토양으로 유입되는 C $l^{-}$, N $O_3$$^{-}$, S $O_4$$^{2-}$ , P $O_4$$^{3-}$ 그리고 유기산 등의 음이온은 점토광물의 결정구조에 포함되지 않기에 관심 대상이다. 본 논문에서는 포화 또는 불포화 상태의 청원통 Bt 층 토양에 특정 또는 비특정적으로 흡착된 음이온을 혼입치환시킴으로서 토양내 자연 황산이온의 용탈과 파쇄곡선을 조사하였다. 시험결과 파쇄곡선 모양과 형태는 토양입자 표면에서의 치환과 흡착에 따라 영향을 받은 것을 알 수 있었다. 포화와 불포화토양분 조건에서 조사된 옥살릭 이온의 파쇄곡선은 토양 자체의 황산이온을 치환시키는데 작은 공극 수량이 필요하였으며 최대 감지한계에 도달은 불포화보다 포화조건에서 더 빨리 도달하였다. 그리고 오른쪽으로 치우친 파쇄곡선은 비록 토양내 유속변화 순에 따라 영향을 받았지만 주로 각각의 음이온의 서로 다른 흡착 특성에 의해 영향을 받았다고 판단된다. 교호상태의 조건에서 얻어진 파쇄곡선과 용출특성은 C/Co가 0보다는 1에 빨리 도달하였다. 이와 같이 음이온의 비대칭형 형태의 용출 특성은 토양을 통과하는 토양수내의 음이온이 전회를 띠고 있는 토양입자 표면과의 선택적 반응에 기인한다. 그리고 토양을 통과하는 용출수의 pH 변화를 조사한 결과 토양 내의 치환과 흡착반응이 진행되는 6 공극수량 까지는 pH가 7까지 증가되나 이 이후 점진적으로 4까지 감소됨을 알 수 있었다.

• 한국환경농학회지
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• 제26권4호
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• pp.294-299
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• 2007

Natural gypsum has been used as a soil amendment in the United States. However, flue gas desulfurization (FGD)-gypsum has not traditionally been used for agricultural purpose although it has potential benefit as a soil amendment. To expand use of FGD-gypsum for agricultural purpose, the effect of FGD-gypsum on soil chemical properties was investigated in the field scales. Application rates for this study were 0 (control), 1.1, and 2.2 Mg ha-1 of FGD-gypsum. After two year application, the soil samples were taken to 110 cm depth and sub-sampled at 10 cm intervals. The heavy metal contents in FGD-gypsum were lower than ceiling levels allowed by regulations for land-applied biosolids. Soil pH was not largely affected by FGD-gypsum application. Although degree of calcium (Ca) saturation in surface horizons increases only slightly with respect to the control, there is a clear decrease in exchangeable aluminum (Al). FGD-gypsum clearly increases the soil electrical conductivity (EC) with increasing application rate. Water-soluble Ca and sulfate is increased with FGD-gypsum application and these ions moved to a depth of at least 80 cm after only 2 years. We conclude that surface application of FGD-gypsum can mitigate toxicity of Al and deficiency of Ca in subsoil of acid soil.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제32회 학술심포지움
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.408-413
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• 2006

A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of $55^{\circ}C$ and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to $60^{\circ}C$ within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were $30^{\circ}C$ and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1449-1459
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• 2015

A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg²⁺ and Ca²⁺ and completely inhibited by Cu²⁺, but slightly enhanced by Fe²⁺. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.