• 대한환경공학회지
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• 제33권11호
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• pp.847-854
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• 2011

주요 신재생에너지인 바이오에너지의 일환으로 조류를 이용한 바이오에너지 및 자원화 기술에 대한 관심이 높아지고 있다. 조류는 영양염류 제거 능력을 활용해서 하수와 같은 오폐수 내 난분해성오염물질과 영양염류 제거의 고도처리도 가능하다. 조류와 박테리아 간의 생태적인 상호작용이 조류를 활용한 하수처리 및 하수자원화에 중요한 역할을 함에도 불구하고, 실지 하수 조건에서 조류와 박테리아간의 생태학적인 상호작용에 관한 과학적인 정보가 부족하다. 본 연구에서는 하수에서 배양이 잘 되고, 지질함량이 높다고 알려진 국내 조류 종인 Ankistrodesmus gracilis SAG 278-2의 하수오염물질 제거 특성과 조류 주입에 따른 하수 박테리아 군집의 반응을 실지 하수 조건에서 연구하였다. 하수 박테리아의 수가 증가는 조류의 성장 속도를 감소시켰으나, 반면 조류의 성장은 박테리아의 생존 및 내성호흡 생분해 속도에는 영향을 주지 않았다. 조류가 주입된 하수에서 난분해성 유기물질 및 총질소의 제거 향상이 관찰되었다. 박테리아 16S rRNA 유전자 T-RFLP 분석에 따르면 조류의 주입은 시간에 따라 박테리아 군집에 영향을 주었다. 박테리아 16S rRNA 유전자 PCR 증폭, clone 및 염기서열 분석 결과, 하수 내 조류의 성장은 박테리아 군집 구성을 변화시키며, 조류와 함께 공동 성장 가능한 박테리아는 Sediminibacterium, Sphingobacterium, Mucilaginibacter 속에 속하는 개체로 판명되었다.

• 대한환경공학회지
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• 제33권5호
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• pp.314-324
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• 2011

하수를 이용해서 배양된 조류는 바이오디젤 생산에 유용한 자원이다. 그러나 실제 하수에서 조류의 영양염류 신진대사와 하수 세균과의 상호작용에 관한 연구는 미흡하다. 본 연구에서는 하수로 배양되는 대표적 조류균주인 Ankistrodesmus gracilis SAG 278-2에 의한 하수 내 질소, 인 제거 거동을 평가하였고, 조류와 상호작용하는 하수 내의 세균 군집을 분석하였다. 하수 슬러지 세균 군집과 비교하였을 때, 조류-세균 복합 군집은 하수 내보다 높은 영양염류 제거를 보였다. 16S rRNA 유전자 분석 결과, 조류-세균 군집에서 조류가 성장함에 따라 Unclassified Alcaligenaceae 세균이 선택적으로 우점됨을 알 수 있었고, 조류에 의해서 선택적으로 우점화된 하수세균은 자연 수질 환경에서 조류와 공생적으로 상호작용 하는 것으로 알려진 Alcaligenes faecalis subsp. 5659-H와 계통학적으로 가까운 것으로 밝혀졌다. 본 연구의 결과, 하수 내의 높은 영양염류 제거를 보이는 조류-세균 복합 군집에서의 조류의 성장 및 신진대사가 특정 세균의 분포에 영향을 주는 것을 알 수 있었다.

• Journal of Plant Biotechnology
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• 제36권4호
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• pp.415-422
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• 2009

This study aimed to develop carnation cultivars with new coloring system. We used four genes of Petunia hybrida - chalcone synthase (CHS), flavanone 3-hydroxylase (FHT), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS) - as probes, in order to isolate four genes from carnations (Dianthus Caryophyllus). The isolated genes were used as probes in order to select mutants out of collected carnations, using Northern blot analysis. The Northern blot analysis revealed 10 DFR mutants - Gumbyul, Eunbyul, Ballatyne, Crystal, Eugenia, Koreno, Imp. White Sim, West Crystal, White Alpine, and White Charotte. Six among the selected 10 cultivarswere excluded from the target cultivars, because Eugenia, Imp. White Sim, and White Alpine were proved to be double mutants of DFR and ANS, Koreno was considered to be a double mutant of DFR and CHS, and Gumbyul and Ballatyne were proved to be double mutants of DFR and CHI (Chalcone isomerase). Consequently, we selected five DFR mutants, including Virginie, which was already selected as a DFR mutant. Finally, we measured DFR activities in order to confirm the selection, and the results showed that all of the five cultivars - Eunbyul, Crystal, West Crystal, White Charotte, and Virginie - had got no DFR activity.

• 한국물환경학회지
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• 제23권6호
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• pp.934-944
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• 2007

Fine suspended solids (SS) induced into a reservoir after flood events play important ecological and water quality roles by presenting persistent turbidity and attenuating light. Thus the origin and physical features must be characterized to understand their transport processes and associated impacts, and for the establishment of watershed based prevention strategies. This study was aimed to characterize the physical properties of the SS sampled from Daecheong Reservoir and its upstream rivers during flood events. Extensive field and laboratory experiments were carried out to identify the turbidity-SS relationships, particle size distributions, settling velocity, and mineral compositions of the SS. Results showed that the turbidity-SS relationships are site-specific depending on the locations and flood events in the system. The turbidity measured within the reservoir was much greater than that measured in the upstream rivers for the same SS value. The effective diameters ($D_{50}$) in the rivers were in the range of $13.3{\sim}54.3{\mu}m$, while those in the reservoir were reduced to $2.5{\sim}14.0{\mu}m$ due to a fast settling of large particles in the rivers. The major minerals consisting of the SS were found to be Illite, Muscovite, Albite, and Quartz both in the rivers and reservoir. Their apparent settling velocities at various locations in the reservoir were in the range of 0.06~0.13 m/day. The research outcome provides a fundamental information for the fine suspended particles that cause persistent turbidity in the reservoir, and can be used as basic parameters for modeling study to search watershed based optimal control measures.

• ALGAE
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• 제25권3호
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• pp.141-153
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• 2010

Seagrass epiphyte blooms potentially have important economic and ecological consequences in Tampa Bay, one of the Gulf of Mexico's largest estuaries. As part of a Tampa Bay pilot study to monitor the impact of environmental stresses, precise characterization of epiphyte diversity is required for efficient management of affected resources. Thus, epiphyte diversity may be used as a rational basis for assessment of ecosystem health. In May 2001, epiphytic species encompassing green, brown and red macroalgae were manually collected from dense and sparse seagrass beds of Thalassia testudinum and Syringodium filiforme. A total of 20 macroalgal epiphytes, 2 Chlorophyta, 2 Phaeophyta, and 16 Rhodophyta, were found on T. testudinum and S. filiforme seagrass at the four sampling sites (Bishop Harbor, Cockroach Bay, Feather Sound, and Mariposa Key). The Rhodophyta, represented by 16 species, dominated the numbers of species. Among them, the thin-crusted Hydrolithon farinosum was the most commonly found epiphyte on seagrass leaves. Species number, as well as species frequency of epiphytes, is higher at dense seagrass sites than sparse seagrass sites. Four attachment patterns of epiphytes can be classified according to cortex and rhizoid development: 1) creeping, 2) erect, 3) creeping & erect, and 4) erect & holding. The creeping type is characterized by an encrusting thallus without a rhizoid or holdfast base. Characteristics of the erect type include a filamentous thallus with or without a cortex, and a rhizoid or holdfast base. The creeping and erect type is characterized by a filamentous thallus with a cortex and rhizoid. A filamentous thallus with a cortex, holdfast base, and host holding branch is characteristics of the erect and holdfast attachment type. This study characterized each species found on the seagrass for epiphyte identification.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.503-508
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• 2011

A ${\beta}$-glucosidase from Phoma sp. KCTC11825BP isolated from rotten mandarin peel was purified 8.5-fold with a specific activity of 84.5 U/mg protein. The purified enzyme had a molecular mass of 440 kDa with a subunit of 110 kDa. The partial amino acid sequence of the purified ${\beta}$-glucosidase evidenced high homology with the fungal ${\beta}$- glucosidases belonging to glycosyl hydrolase family 3. Its optimal activity was detected at pH 4.5 and $60^{\circ}C$, and the enzyme had a half-life of 53 h at $60^{\circ}C$. The $K_m$ values for p-nitrophenyl-${\beta}$-D-glucopyranoside and cellobiose were 0.3 mM and 3.2 mM, respectively. The enzyme was competitively inhibited by both glucose ($K_i$=1.7 mM) and glucono-${\delta}$-lactone ($K_i$=0.1 mM) when pNPG was used as the substrate. Its activity was inhibited by 41% by 10 mM $Cu^{2+}$ and stimulated by 20% by 10 mM $Mg^{2+}$.

• Mycobiology
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• 제40권3호
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• pp.173-180
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• 2012

A ${\beta}$-glucosidase from Penicillium italicum was purified with a specific activity of 61.8 U/mg, using a chromatography system. The native form of the enzyme was an 88.5-kDa tetramer with a molecular mass of 354 kDa. Optimum activity was observed at pH 4.5 and $60^{\circ}C$, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and $65^{\circ}C$, respectively. Its activity was inhibited by 47% by 5 mM $Ni^{2+}$. The enzyme exhibited hydrolytic activity for p-nitrophenyl-${\beta}$-D-glucopyranoside (pNP-Glu), p-nitrophenyl-${\beta}$-D-cellobioside, p-nitrophenyl-${\beta}$-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-${\beta}$-D-lactopyranoside, p-nitrophenyl-${\beta}$-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a ${\beta}$-glucosidase. The $k_{cat}/K_m\;(s^{-1}mM^{-1})$ values for pNP-Glu and cellobiose were 15,770.4 mM and 6,361.4 mM, respectively. These values were the highest reported for ${\beta}$-glucosidases. Non-competitive inhibition of the enzyme by both glucose ($K_i=8.9mM$) and glucono-${\delta}$-lactone ($K_i=11.3mM$) was observed when pNP-Glu was used as the substrate. This is the first report of non-competitive inhibition of ${\beta}$-glucosidase by glucose and glucono-${\delta}$-lactone.

• Journal of Ecology and Environment
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• 제46권4호
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• pp.282-291
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• 2022

Background: The Arctic permafrost stores enormous amount of carbon (C), about one third of global C stocks. However, drastically increasing temperature in the Arctic makes the stable frozen C stock vulnerable to microbial decomposition. The released carbon dioxide from permafrost can cause accelerating C feedback to the atmosphere. Soil organic matter (SOM) composition would be the basic information to project the trajectory of C under rapidly changing climate. However, not many studies on SOM characterization have been done compared to quantification of SOM stocks. Thus, the purpose of our study is to determine soil properties and molecular compositions of SOM in four different Arctic regions. We collected soils in different soil layers from 1) Cambridge Bay, Canada, 2) Council, Alaska, USA, 3) Svalbard, Norway, and 4) Zackenberg, Greenland. The basic soil properties were measured, and the molecular composition of SOM was analyzed through pyrolysis-gas chromatography/mass spectrometry (py-GC/MS). Results: The Oi layer of soil in Council, Alaska showed the lowest soil pH and the highest electrical conductivity (EC) and SOM content. All soils in each site showed increasing pH and decreasing SOC and EC values with soil depth. Since the Council site was moist acidic tundra compared to other three dry tundra sites, soil properties were distinct from the others: high SOM and EC, and low pH. Through the py-GC/MS analysis, a total of 117 pyrolysis products were detected from 32 soil samples of four different Arctic soils. The first two-axis of the PCA explained 38% of sample variation. While short- and mid-hydrocarbons were associated with mineral layers, lignins and polysaccharides were linked to organic layers of Alaska and Cambridge Bay soil. Conclusions: We conclude that the py-GC/MS results separated soil samples mainly based on the origin of SOM (plants- or microbially-derived). This molecular characteristics of SOM can play a role of controlling SOM degradation to warming. Thus, it should be further investigated how the SOM molecular characteristics have impacts on SOM dynamics through additional laboratory incubation studies and microbial decomposition measurements in the field.

• Asian-Australasian Journal of Animal Sciences
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• 제31권1호
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• pp.63-70
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• 2018

Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

• 환경생물
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• 제39권4호
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• pp.445-451
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• 2021

Agrobacterium tumefaciens strain CP4 유래 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) 유전자를 포함하는 유전자변형생물체(Living modified organism, LMO)가 개발되었다. 이 같은 LMO는 국내 승인되어 사료용, 식품용, 가공용으로 이용 중이다. 간이면역 검사키트 개발을 위해서는 고효율의 단클론 항체 개발이 필수적이다. 본 연구에서는 대장균 BL21 (DE3)에서 재조합 CP4 EPSPS 단백질을 정제하였으며 SDS-PAGE와 MALDI-TOF MS 분석으로 단백질 특성을 분석하였다. 단클론 항체 제작은 (주)앱클론의 SOP 매뉴얼에 따라 진행하였다. 본 연구 결과 5개의 단클론 항체 클론(2F2, 4B9, 6C11, 10A9, 10G9)를 확보하였다. 5종의 단클론 항체의 효율과 특이도 검정을 위해서 LM 면화 추출액을 이용한 western blotting 분석을 실시하였다. 모든 단클론 항체는 CP4 EPSPS를 함유하는 MON1445와 MON88913을 특이적으로 검출하였으며 비변형 면화 및 타종의 LM 면화에서는 검출되지 않았다. 이러한 결과들을 바탕으로 CP4 EPSPS 단클론 항체는 LMO에 함유된 CP4 EPSPS 단백질을 타겟으로 항체 기반 검출법 개발에 활용될 것으로 사료된다.