• Title/Summary/Keyword: Residues

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Cloning and Sequencing of the pelCl Gene Encoding Pectate Lyase of Erwinia carotovora subsp. carotovora LY34 (Erwinia carotovora subsp. carotovora LY34에서 pelCI 유전자 클로닝)

  • Lim, Sun-Tech;Park, Yong-Woo;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.380-387
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    • 1997
  • Phytopathogenic Erwinia carotovora subsp. carotovora (Ecc) LY34 causes plant tissue maceration by secretion of pectinolytic enzymes such as pectate Iyase (PL) existed as multiple isoenzyme form. Genomic DNA from Ecc LY34 was digested with Sau3Al and ligated into the BamHI site of pBluescript ll $SK^+$. Among them, a clone hydrolyzing polypectate was selected and its DNA was digested with BamHI. Through the subsequent subcloning the resulting 3.1 kb fragment, corresponding to a peICI, was subcloned into pLYPA 100. The structural organization of a peICI gene encoding a 374 amino acid residues consists of an open reading frame (ORF) of 1,122 bp commencing with a ATG start codon and followed by a TAA stop codon. PeICI contained a typical prokaryotic signal peptide of 22-amino acid. Since the deduced amino acid sequences of PeICl protein was very similar to those of PelIII of Erwinia carotovora subsp. carotovora, and to those of Pel3 of Erwinia carotovora subsp. atroseptica, and to those of PeIC of Erwinia carotovora subsp. carotovora, it belong to the same family PLbc group. The 374-amino acld PeICI had a calculated Mr of 40,507 and pI of 7.60.

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Biotinoyl Domain of Human Acetyl-CoA Carboxylase;Structural Insights into the Carboxyl Transfer Mechanism

  • Lee, Chung-Kyung;Cheong, Hae-Kap;Ryu, Kyoung-Seok;Lee, Jae-Il;Jeon, Young-Ho;Cheong, Chae-Joon
    • Journal of the Korean Magnetic Resonance Society
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    • v.12 no.1
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    • pp.1-13
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    • 2008
  • Acetyl-CoA carboxylase (ACC) catalyzes the first step in fatty acid biosynthesis: the synthesis of malonyl-CoA from acetyl-CoA. As essential regulators of fatty acid biosynthesis and metabolism, ACCs are regarded as therapeutic targets for the treatment of metabolic diseases such as obesity, In ACC, the biotinoyl domain performs a critical function by transferring an activated carboxyl group from the biotin carboxylase domain to the carboxyl transferase domain, followed by carboxyl transfer to malonyl-CoA. Despite the intensive research on this enzyme, only the bacterial and yeast ACC structures are currently available, To explore the mechanism of ACC holoenzyme function, we determined the structure of the biotinoyl domain of human ACC2 and analyze its characteristics using NMR spectroscopy. The 3D structure of the hACC2 biotinoyl domain has a similar folding topology to the previously determined domains from E. coli and P. Shermanii, however, the 'thumb' structure is absent in the hACC2 biotinoyl domain. Observations of the NMR signals upon the biotinylation indicate that the biotin group of hACC2 does not affect the structure of the biotinoyl domain, while the biotin group for E. coli ACC interacts directly with the thumb residues that are not present in the hACC2 structure. These results imply that, in the E. coli ACC reaction, the biotin moiety carrying the carboxyl group from BC to CT can pause at the thumb of the BCCP domain. The human biotinoyl domain, however, lacks the thumb structure and does not have additional non-covalent interactions with the biotin moiety; thus, the flexible motion of the biotinylated lysine residue must underlie the "swinging arm" motion. This study provides insight into the mechanism of ACC holoenzyme function and supports the "swinging arm" model in human ACCs.

A Study on Environmental Monitoring of Fluorouracil and Decontamination Reagents (Fluorouracil의 환경감시 및 제거약제에 관한 연구)

  • Lee, Su-Gil;Lee, Nae-Woo
    • Journal of the Korean Society of Safety
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    • v.24 no.4
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    • pp.47-52
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    • 2009
  • This study has been to examine the occupational exposure levels of Fluorouracil (5-FU) in a hospital and to investigate the most effective cleaning reagent for control. Fluorouracil is one of the cytotoxic drugs which are therapeutic agents used to treat cancer. The health practitioners working in the cytotoxic work room and oncology ward areas are exposed to adverse health risks like cytogenetic and DNA damage from cytotoxic drugs exposure by frequent skin contact from contaminated surfaces. Four kinds of cleaning reagents has been examined to degrade the 5-FU. It was found that 5-FU was only degraded soon after the reaction in 0.5%(w/v) NaClO solution. Therefore, 0.5%(w/v) NaClO solution has been chosen to decompose any residues on the contamination surfaces. A substantial level of contamination was found on the surfaces of cytotoxic work room and oncology ward areas. The contamination ranges of the surfaces in cytotoxic work room and oncology ward areas were from 2.0 to $13.8{\mu}g/m^2$ and 5.39 to $11.53{\mu}g/m^2$ respectively. Consequently, regulation of the occupational exposure limit, procedure of special cleaning, and the use of personal protective equipment are recommended during the manipulation and administration of the drugs to avoid skin contamination from cytotoxic drugs like 5-FU.

Silicidation Reaction Stability with Natural Oxides in Cobalt Nickel Composite Silicide Process (자연산화막 존재에 따른 코발트 니켈 복합실리사이드 공정의 안정성)

  • Song, Oh-Sung;Kim, Sang-Yeob;Kim, Jong-Ryul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.8 no.1
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    • pp.25-32
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    • 2007
  • We investigated the silicide reaction stability between 10 nm-Col-xNix alloy films and silicon substrates with the existence of 4 nm-thick natural oxide layers. We thermally evaporated 10 nm-Col-xNix alloy films by varying $x=0.1{\sim}0.9$ on naturally oxidized single crystal and 70 nm-thick polycrystalline silicon substrates. The films structures were annealed by rapid thermal annealing (RTA) from $600^{\circ}C$ to $1100^{\circ}C$ for 40 seconds with the purpose of silicidation. After the removal of residual metallic residue with sulfuric acid, the sheet resistance, microstructure, composition, and surface roughness were investigated using a four-point probe, a field emission scanning electron microscope, a field ion bean4 an X-ray diffractometer, and an Auger electron depth profiling spectroscope, respectively, to confirm the silicide reaction. The residual stress of silicon substrate was also analyzed using a micro-Raman spectrometer We report that the silicide reaction does not occur if natural oxides are present. Metallic oxide residues may be present on a polysilicon substrate at high silicidation temperatures. Huge residual stress is possible on a single crystal silicon substrate at high temperature, and these may result in micro-pinholes. Our results imply that the natural oxide layer removal process is of importance to ensure the successful completion of the silicide process with CoNi alloy films.

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Evaluating The Fuel Characteristics of Wood Pellets Fabricated with Wood Tar and Starch as An Additive (목타르와 전분 첨가제 혼합에 따른 목재펠릿 품질특성 평가)

  • Ahn, Byoung-Jun;Lee, Soo-Min
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.3
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    • pp.318-326
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    • 2014
  • This study was conducted to investigate the potential of non-used forest biomass residues as raw materials for making wood pellets with additives such as wood tar and starch and to evaluate fuel characteristics of the pellets. Wood tar, a by-product provided from the carbonization process of wood, could be a suitable additive for wood pellet production due to its higher calorific value and lower hazardous heavy metals, such as cadmium and mercury, compared to woody biomass. When the wood tar (10 wt%) was added, the calorific value was increased from 4,630 kcal/kg (wood pellet without additive) to 4,800 kcal/kg (wood pellet with additive). With the increase of additive amount into wood pellet, the length and individual density of wood pellet increased. In addition, bulk density of the pellets was increased, whereas the fine content was decreased. Consequently the overall productivity of wood pellets was improved by adding 2 w% additives into wood pellets; the percentage of productivity increase was 5.9% and 4.9% for adding starch and wood tar, respectively.

Germ Line Transformation of the Silkworm, Bombyx mori L. with a piggyBac Vector Harboring the Human Lactoferrin Gene (락토페린 유전자도입 piggyBac 벡터에 의한 누에 형질전환)

  • Kim, Yong-Soon;Sohn, Bong-Hee;Kim, Kee-Young;Jung, I-Yeon;Kim, Mi-Ja;Kang, Pil-Don
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.37-42
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    • 2007
  • Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

Assessment of Allelopathic Potential of Some Weed Species on Alfalfa(Medicago sativa L.) Germination and Early Seedling Growth (알팔파 발아와 초기생육에 대한 잡초종의 Allelopathic 잠재성 평가)

  • Chung, I.M.;Miller, D.A.
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.121-130
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    • 1995
  • Greenhouse and laboratory studies were conducted to investigate allelopathic potential of some weed species on alfalfa(Medicago sativa L.) germination and seedling growth. In the comparison between top(leaves+stems) and root extracts, top extract exhibited greater allelopathic effects on alfalfa germination than that of root. The various weed species extract differently responded to alfalfa test species, WL-320, in terms of allelopathic effect. Top and root aqueous extracts of lambsquarter(Chenopodium album L.), giant foxtail(Setaria faberii Herrm.), redroot pigweed (Amaranthus retroflexus L.), velvetleaf(Abutilon theophrasti Medic.), crabgrass(Digitaria sanguinalis L.), canada thistle(Cirsium arvense L.) and prostrate knotweed(Polygonium aviculare L.) significantly inhibited germination, seedling length, weight, vigor, and rate of germination of alfalfa. The regression slopes of various top extracts showed that velvetleaf(b=3.69) extracts were the most inhibitory, while large crabgrass(b=2.39) extracts had the least allelopathic effect on alfalfa germination. Germination, seedling length and weight of alfalfa were inversely proportional to the concentration of dried velvetleaf extracts. Also, more of the toxic effects were observed from the dried extracts compared to the fresh extracts. Residue of velvetleaf inhibited significantly alfalfa emergence and survival percentage compared to the control. The emergence and survival percentage of alfalfa were 44%, 57% at 1.0% residue treatment, respectively. When weed residues were mixed with silica sand with incubation time, velvetleaf residue most inhibited alfalfa growth. The degree of inhibition increased as incubation time increased. An incubation for 72h caused the greatest inhibition of alfalfa growth. These results demonstrate the different allelopathic activity of weed species extracts on alfalfa and suggest that weed may affect alfalfa growth and development through the inhibitory effects of allelochemicals present in weed tissue.

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The Development of Dimerized Chicken Recombinant Single-chain Fv (ScFv) Antibody Using Leucine Zipper Motif (Leucine Zipper Motif를 이용한 닭의 재조합 이량체 Single-chain Fv (ScFv) 항체의 개발)

  • Park, Dong-Woon;Kim, Eon-Dong;Kim, Sung-Heon;Han, Jae-Yong;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.328-334
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    • 2011
  • Leucine zipper motif consists of multiple periodic leucine residues, which forms amphipathic alpha helix. The hydrophobic nature of leucine zipper motif can dimerize proteins which contain this motif. Leucine zipper motif addition at C-terminus of single-chain Fv (ScFv) antibody induces its dimerization. Since the dimeric ScFv antibody contains two antigen binding sites (bivalency) like Y-shaped complete antibody, it could increase avidity. As a result, it could show higher antigen binding activity than monomeric ScFv antibodies. Based on this concept, monomeric chicken 8C3 ScFv antibody previously developed from chicken hybridoma was dimerized by the addition of leucine zipper motif at C-terminus of ScFv antibody. The dimeric 8C3 ScFv antibody specifically reacted with Eimerian sporozoite which causes Avian Coccidiosis. As expected, dimeric 8C3 ScFv antibody showed 3-folds higher antigen binding activity than monomer due to increased avidity. In addition, protien yields of dimer expression were 2-folds higher than monomer.

A Study on the Characteristics of Humic Materials Extracted from Decomposing Plant Residues -III. Amino Acids in the Acid Hydrolysates of Humic Acids Extracted from Straw of Rice and Barley (식물성(植物性) 유기질(有機質)의 부숙과정중(腐熟過程中) 부식특성(腐植特性)에 관(關)한 연구(硏究) -III. 볏짚과 보리짚부식산(腐植酸)의 산가수분해(酸加水分解) 용액중(溶液中) Amino 산(酸)의 함량(含量))

  • Kim, Jeong-Je;Lee, Wi-Young;Shin, Young-Oh
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.301-306
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    • 1988
  • Contents and distribution of amino acids in the hydrolysates of humic acids extracted from straw of rice and barley at three different dates during decomposition were examined. The results obtained from this study may be summed up as the following: 1. There are differences between the humic acid hydrolysates from rice straw and barley straw in regards of composition of humic acids and distribution of amino acids. 2. Neutral amino acids as a group occupy the largest share, followed by acidic amino acids and basic amino acids. 3. The total amount of amino acids per gram of humic acid is greater in straw of rice than in straw of barley. 4. With the humification progressing the content of lysine increases, but the content of histidine decreases. In general glycine, glutamic acid, aspartic acid, alanine and leucine constitute the 5 predominant amino acids in all hydrolysates. 5. Arginine is not detected at all in any of the hydrolysates of humic acids obtained from humified materials. 6. The presence of phenylalanine and tyrosine is an evidence for the aromatic characteristics of humic acids.

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Partial Characterization of Two Cathepsin D Family Aspartic Peptidases of Clonorchis sinensis

  • Kang, Jung-Mi;Yoo, Won-Gi;Le, Huong Giang;Thai, Thi Lam;Hong, Sung-Jong;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
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    • v.57 no.6
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    • pp.671-680
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    • 2019
  • Cathepsin D (CatD, EC 3.4.23.5) is a member belonging to the subfamily of aspartic endopeptidases, which are classified into the MEROPS clan AA, family A1. Helminth parasites express a large set of different peptidases that play pivotal roles in parasite biology and pathophysiology. However, CatD is less well known than the other classes of peptidases in terms of biochemical properties and biological functions. In this study, we identified 2 novel CatDs (CsCatD1 and CsCatD2) of Clonorchis sinensis and partially characterized their properties. Both CsCatDs represent typical enzymes sharing amino acid residues and motifs that are tightly conserved in the CatD superfamily of proteins. Both CsCatDs showed similar patterns of expression in different developmental stages of C. sinensis, but CsCatD2 was also expressed in metacercariae. CsCatD2 was mainly expressed in the intestines and eggs of C. sinensis. Sera obtained from rats experimentally infected with C. sinensis reacted with recombinant CsCatD2 beginning 2 weeks after infection and the antibody titers were gradually increased by maturation of the parasite. Structural analysis of CsCatD2 revealed a bilobed enzyme structure consisting of 2 antiparallel β-sheet domains packed against each other forming a homodimeric structure. These results suggested a plausible biological role of CsCatD2 in the nutrition and reproduction of parasite and its potential utility as a serodiagnostic antigen in clonorchiasis.