• Title/Summary/Keyword: active residues

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Fates of Cyfluthrin and Trichlorfon in Water and Their Impacts on Aquatic Organisms Following Aerial Application Over the Forest (삼림환경에 항공살포된 Cyfluthrin과 Trichlorfon의 물에서의 동태와 수서생물에 미치는 영향)

  • Lee, Sung-Kyu;Kim, Yong-Hwa;Kim, Tae-Wook;Roh, Jung-Koo
    • Korean Journal of Environmental Agriculture
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    • v.8 no.1
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    • pp.17-29
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    • 1989
  • This study was conducted to evaluate the behavior in the water and the impact on aquatic organisms following aerial application of two insecticides in the forest, cyfluthrin and trichlorfon, to control the alder leaf beetle. As active ingredients, 25g of cyfluthrin and 536g of trichlorfon per ha were diluted seperately into 30L of tap water, and applied with a helicopter to the study areas. A model stream study was also conducted in a stream located adjacent to the study area in order to confirm the impact of insecticides on aquatic invertebrates. Cyfluthrin residues in water were $0.62{\mu}g/L$ (1st. application) and $78{\mu}g/L$ (2nd application) immediately after spraying. and decreased, to a non-detectable level after one day, while trichlorfon residue increased to $30.7{\mu}g/L$ one day after spraying and fluctuated for 22th day depending on precipitation after spraying. Cyfluthrin application rapidly increased the number of some drifting aquatic invertebrates during 24-hour period immediately after spraying, but had no effects on the other aquatic organisms such as fish and zooplankton. The largest increase in the number of drifting organisms following application of cyfluthrin was shown by Ephemeroptera, and followed by Trichoptera, Coleoptera, and Diptera. However, trichlorfon little affected the number of drifting aquatic invertebrates and zooplankton population.

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Molecular Cloning of Glycoside Hydrolase Family 74 Genes and Analysis of Transcript Products from the Basidiomycete Phanerochaete chrysosporium (담자균 Phanerochaete chrysosporium으로부터 유래한 Glycoside Hydrolase Family 74 유전자 클로닝과 전사산물 분석)

  • Lee, Jae-Won;Samejima, Masahiro;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.3
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    • pp.56-63
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    • 2006
  • In order to evaluate the mechanism of cellulose hydrolysis, the complementary DNA encoding Glycoside Hydrolase Family (GHF)74 was cloned from Phanerochaete chrysosporium. Depending on the presence of Cellulose Binding Module (CBM), it can be classified as GHF74A or GHF74B. The GHF74A gene from P. chrysosporium (PcGHF74A) consists of 2163 bp encoding a protein of 721 amino acid residues. The PcGHF74A showed homology of 70~77% compared with the GHF74 from other filamentous fungi. The PcGHF74B, which contains CBM and is a member of family 1, was transcribed to various transcripts depending on the nature of carbon sources and their concentration. To study the possible presence of splice variants in GHF74B transcripts in P. chrysospoium, we carried out RT-PCR analysis using primers that designed based on the annotation data and sequenced data. Our result indicated that PcGHF74B was transcribed to several splicing variants in various culture conditions. Especially in the culture of 2% cellulose, three transcript products were observed. First transcript was presumed to be a full length ORF that contained 11th intron with stop codon at position 2562 bp. The second one consisted of 12 exons and 11 introns with stop codon at position 1187 bp with 7th exon. The shortest transcript consisted of 10 exons and 9 introns with stop codon at 910 bp in the 7th exon. These premature stop codon might prevent the synthesis of fully active GHF74 or contribute for the production of protein with distinct function depending on the ambient carbon sources.

Characterization of Barley ${\alpha}$-Amylase Chimeric Enzymes Expressed in Pichia pastoris (Pichia pastoris에서 발현된 보리 알파아밀라제 Chimera 효소들의 특성)

  • Kim, Tae-Jip;Yuk, Jeong-Bin;Choi, Seung-Ho;Jang, Myoung-Uoon;Svensson, Birte
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.80-85
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    • 2010
  • Two different ${\alpha}$-amylase isozymes (AMY1 and AMY2) found in barley malt share up to 80% of amino acid sequence identity with each other, but their enzymatic properties differ remarkably. AMY1 shows the highest activity at low concentration of calcium ion, while AMY2 is highly active at high calcium concentration. Meanwhile, BASI (Barley ${\alpha}$-Amylase/Subtilisin Inhibitor) protein specifically inhibits only AMY2. In the present study, three separate regions in AMY genes (I, II, and III) were assigned on the basis of restriction enzyme sites and four kinds of chimeric amylases have been obtained by swapping a part of regions with each other. Each chimera gene was successfully over-expressed in Pichia pastoris. From the results of enzymatic characterization, both AMY211 and AMY122 showed the mixed or intermediate type of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY221 chimera could be significantly inhibited by BASI protein. As a result, it can be proposed that some amino acid residues in the region I and II, except region III, of barley ${\alpha}$-amylases play very important roles in calcium-dependency and interaction with BASI.

The Role of Geocrete and Soluble Sodium Silicate as a Substitute to Control Leachate Leaking from Landfill Side Wall (Geocrete와 규산소다액을 이용한 매립지 사면 침출수 누출제어)

  • 조재범;현재혁;나진성;김자영
    • Journal of the Korean Geotechnical Society
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    • v.17 no.6
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    • pp.47-51
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    • 2001
  • There are two strategies to cope with the troubles in landfill site after closure. The first method is active in a way that the wastes are dug up and the recyclable materials are reutilized, meanwhile the materials not recyclable are incinerated in order to minimize the volume of residues to be disposed of. The second method is rather passive and defensive in a way that the source of contamination, that is, buried wastes are not treated. Instead, the transport of leaking leachate and gases generated from the wastes are intercepted and controlled. In this study, as a passive way of the efficient leachate blocking process, applicabilities of geocrete and soluble sodium silicate as a substitute to control leachate leaking from landfill sidewall were investigated. In case of compression test, the strength of mixture I (Geocrete:Sodium silicate=1:3.9 v/v) and mixture II (Geocrete:Sodium silicate=1:2.5 v/v), even after 7 days' curing was higher than the minimum allowance to tolerate the loading(5 kg/$\textrm{cm}^2$). Soaking in the acid fur 4 days and 7 days respectively, the compressive strength of the specimens reduced seriously. The toxicity of geocrete is not detected through the bioassay test, once it was mixed with sodium silicate and the complex was formed. The hydraulic conductivity of the mixtures even after 7 days' curing was lower than the threshold limit $(1.0\times10_{-7}cm/s)$.

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Gene Cloning and Characterization of a Cold-Adapted Esterase from Acinetobacter venetianus V28

  • Kim, Young-Ok;Heo, Yu Li;Kim, Hyung-Kwoun;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Dong-Gyun;Kim, Woo-Jin;Kim, Bong-Seok;Jee, Young-Ju;Lee, Sang-Jun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1245-1252
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    • 2012
  • Acinetobacter venetians V28 was isolated from the intestine of righteye flounder, Poecilopsetta plinthus caught in Vietnam seawater, and the esterase gene was cloned using a shotgun method. The amino acid sequence deduced from the nucleotide sequence (1,017 bp) corresponded to a protein of 338 amino acid residues with a molecular weight of 37,186. The esterase had 87% and 72% identities with the lipases of A. junii SH205 and A. calcoaceticus RUH2202, respectively. The esterase contained a putative leader sequence, as well as the conserved catalytic triad (Ser, His, Asp), consensus pentapeptide GXSXG, and oxyanion hole sequence (HG). The protein from the strain V28 was produced in both a soluble and an insoluble form when the Escherichia coli cells harboring the gene were cultured at $18^{\circ}C$. The maximal activity of the purified enzyme was observed at a temperature of $40^{\circ}C$ and pH 9.0 using p-NP-caprylate as substrate; however, relative activity still reached to 70% even at $5^{\circ}C$ with an activation energy of 3.36 kcal/mol, which indicated that it was a cold-adapted enzyme. The enzyme was a nonmetallo-protein and was active against p-nitrophenyl esters of $C_4$, $C_8$, and $C_{14}$. Remarkably, this enzyme retained much of its activity in the presence of commercial detergents and organic solvents. This cold-adapted esterase will be applicable as catalysts for reaction in the presence of organic solvents and detergents.

Persistent Organic Pollution and Arsenic Contamination in Asia Pacific Water: Case Study of Emerging Environmental Problems in Vietnam

  • Pham, Viet.H.
    • Journal of Wetlands Research
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    • v.9 no.1
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    • pp.79-89
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    • 2007
  • This paper provides a comprehensive overview of the present status of several environmental problems caused by emerging toxic substances such as persistent organic pollutants (POPs), endocrine disrupting chemicals (EDCs), and arsenic in various environmental media in Vietnam. Monitoring data reported during the 1990s demonstrated elevated contamination of DDTs in most of these compartments in Vietnam. Studies in frame of the Asia-Pacific Mussel Watch Program revealed that fish, mussels and resident birds from Vietnam contained higher concentrations of DDTs as compared to other countries in region, suggesting the role of Vietnamese environment as a significant emission source of DDT in the Southeast Asian region. The estimated dietary intakes of PCBs and DDTs for Vietnamese were relatively high among Asian developing countries, suggesting potential risk for humans posed by thesechemicals. Widespread contamination of some endocrine active compounds such as alkylphenols and phthalates was observed at various sites along the coasts of northern and middle Vietnam. The presence of significant source of bisphenol-A along Red River estuary was revealed with the concentrations comparable to those reported for developed nations. A case study on seasonal variation of alkylphenols and phthalates in surface water of river delta and estuary of north and middle Vietnam indicated the differences in distribution of these compounds between dry and rainy seasons. Higher concentrations of alkylphenols and phthalates were found in dry season in estuary; while the contrasting pattern was observed in the river delta, showing elevated residues in rainy season. This result suggests the different behavior of alkylphenols and phthalates in river delta and coastal environment. From ecotoxicological perspectives, concentrations of bis-phenol A and di(2-ethylhexyl)phthalates [DEHP] in surface water from some locations in Vietnam exceeded the guideline values for Ecotoxicological Effects and the Environmental Risk Limit, respectively, suggesting potential for toxic implications on aquatic wildlife. Widespread and elevated arsenic contamination was discovered inour recent surveys in groundwater in a large area of suburban areas of Hanoi city, the capital of Vietnam. The most recent investigation in 4 villages showed about more than 50 % of groundwater samples contained As concentrations exceeding 50 g/L (the WHO and Vietnamese standard). In particular, in Son Dong villages, 58 % of samples analyzed contained As concentrations higher than 200 g/L. Good correlations were found in As concentrations in water and hair and urine of peoples in corresponding families, suggesting the chronic exposure to As by people living in As-contaminated ground water areas. In Son Dong village, As levels in hair (mean: 1.7 mg/kg dry wt) and urine (g/g creatinine) exceeding the reference values recommended by WHO, suggesting potential for human risk posed by long term accumulation of As in human body. Future studies should be focused on the time trends of POPs and EDCs in biota in Vietnam in order to predict future trend of contamination and to reveal new clues for understanding possible toxic impacts on aquatic organisms. The issues of arsenic contamination in groundwater and their chronic toxic implications on human health should be systematically investigated in the future.

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Effect of Dietary Fiber on the In Vitro Digestibility of Fish Protein (식이 섬유소가 어류단백 소화율에 미치는 영향)

  • Ryu, Hong-Soo;Park, Nam-Eun;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.3
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    • pp.255-262
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    • 1992
  • In vitro digestibility of filefish, protein was substantially decreased by fiber constituents in the follow-ing order : pectin (9.97%), gum karaya (7.03%), sodium alginate (6.12%),and cellulose (1.52%). The order of reduction by fibrous residues from vegetables ranked as follows : sea tangle (12.36%), Ro-maine lettuce (11.12%), perillar leaf (8.96%), and green pepper (5.15%). The inhibitory effect of the dietary fibers towards filefish protein digestion, expressed as soybean trypsin inhibitor equivalents, in-creased with added levels, but the inhibition differed with the sources of dietary fibers. Sea tangle and sodium alginate were most active in decreasing the concentration of essential amino acid from filefish protein hydrolysis. Sodium alginate exerted an inhibitory effect on the activity of trypsin, but the other fiber constituents did not have an inhibitory potency on trypsin and bacterial pretense (Streptomyces griceus). Results supported that dietary fiber components may reduce protein digestibility through the interaction of dietary fiber components with filefish protein.

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Molecular Modification of Perilla Lipid Composition

  • Hwang, Young-Soo;Kim, Kyung-Hwan;Hwang, Seon-Kap;Lee, Sun-Hwa;Lee, Seong-Kon;Kim, Jung-Bong;Park, Sang-Bong;Tom Okita;Kim, Donghern
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.20-30
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    • 1999
  • In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

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Cloning and Characterization of a Novel Carboxylesterase Gene from Cow Rumen Metagenomic Library (소 반추위 메타게놈에서 새로운 carboxylesterase 유전자 클로닝 및 유전산물의 특성)

  • Asraful Islam, Shah Md.;Kim, Min-Keun;Renukaradhya, K. Math;Srinivasa, Reddy R.N.;Kim, Eun-Jin;Kim, Jung-Ho;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1306-1313
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    • 2010
  • The gene encoding esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est1R) was 2,465 bp in length, encoding a protein of 366 amino acid residues, and the molecular weight of the enzyme was 61,166 Da. Est1R of rumen cosmid library shared 5.9% amino acid identity with Est1R (P37967) of PNB carboxylesterase, 6.1% with Est1R (1EEAA) of acetylcholinesterase and 6.1% with Est1R (1H23A) of chain A. BlastP in NCBI database analysis of Est1R revealed that it was not homologous to previous known lipases and esterases. Est1R showed optimum activity at pH 7.0 and $40^{\circ}C$. On the other hand, the enzyme was found to be most active without organic solvent, followed by 95% activity with methanol, and the enzyme activity was highly affected by hexane (lost 51% activity). Therefore, the novel esterase gene est1R is likely obtainable from cow rumen metagenome and may be utilized for industrial purposes.

Cloning and Expression of an Insecticidal Crystal Protein CryIIA Gene from Bacillus thuringiensis subsp. kurstaki HD-1 (Bacillus thuringiensis subsp. kurstaki HD-1 CryIIA의 내독소 단백질 유전자의 클로닝 및 발현)

  • 김호산;김상현;제연호;유용만;서숙재;강석권;조용섭
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.300-306
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    • 1993
  • The CryIIA gene encoding the insecticidal crystal protein of Bacillus thuringiens!s subsp. kurstalri HD-l has been cloned in Escherichia col!, and its nucleotide sequences were determined completely. 5kb Hindlli fragment harboring CryIIA gene was screened in the large ca. 225kb plasmid DNA by southern blot. HindlIT digested 5kb fragment was ligated into pUC19 and transformed in E. coli. The 4kb BamHI-HindlIT fragment containing the CryIIA gene was subcloned and named pSKIIA. DNA sequence analysis demonstrates that pSKIIA is the gene of an operon which is comprised of Lhree open reading frames (designated orn, orf2 and or£3). The CrylIA gene is composed of 3,952bp-long BamHI-Hindill DNA restriction fragment. The orf3 code for a polypeptide of 633 amino acid residues. The protoxin protein has a predicted molecular weight of 70,780. The E. coli derived protoxin gene product is biologICally active against three species of Lepidopteran (Plu.lelia maculipennis, He/iolhis assulta, Spodoptera litura) and a species of Dip Leran( Culex pipines) larvae in bioassay.

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