• Title/Summary/Keyword: Metal binding

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Characterization, Cloning and Expression of the Ferritin Gene from the Korean Polychaete, Periserrula leucophryna

  • Jeong Byeong Ryong;Chung Su-Mi;Baek Nam Joo;Koo Kwang Bon;Baik Hyung Suk;Joo Han-Seung;Chang Chung-Soon;Choi Jang Won
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.54-63
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    • 2006
  • Ferritin is a major eukaryotic protein and in humans is the protein of iron storage. A partial gene fragment of ferritin (255 bp) taken from the total RNA of Periserrula leucophryna, was amplified by RT-PCR using oligonucleotide primers designed from the conserved metal binding domain of eukaryotic ferritin and confirmed by DNA sequencing. Using the $^{32}P-labeled$ partial ferritin cDNA fragment, 28 different clones were obtained by the screening of the P. leucophryna cDNA library prepared in the Uni-ZAP XR vector, sequenced and characterized. The longest clone was named the PLF (Periserrula leucophryna ferritin) gene and the nucleotide and amino acid sequences of this novel gene were deposited in the GenBank databases with accession numbers DQ207752 and ABA55730, respectively. The entire cDNA of PLF clone was 1109 bp (CDS: 129-653), including a coding nucleotide sequence of 525 bp, a 5' -untranslated region of 128 bp, and a 3'-noncoding region of 456 bp. The 5'-UTR contains a putative iron responsive element (IRE) sequence. Ferritin has an open reading frame encoding a polypeptide of 174 amino acids including a hydrophobic signal peptide of 17 amino acids. The predicted molecular weights of the immature and mature ferritin were calculated to be 20.3 kDa and 18.2 kDa, respectively. The region encoding the mature ferritin was subcloned into the pT7-7 expression vector after PCR amplification using the designed primers and included the initiation and termination codons; the recombinant clones were expressed in E. coli BL21(DE3) or E. coli BL21(DE3)pLysE. SDS-PAGE and western blot analysis showed that a ferritin of approximately 18 kDa (mature form) was produced and that by iron staining in native PAGE, it is likely that the recombinant ferritin is correctly folded and assembled into a homopolymer composed of a single subunit.

Lead Biosorption by Alginate Beads Immobilizing Aspergillus niger (Aspergillus niger를 고정화한 Alginate Bead에 의한 납 흡착)

  • Bang, Byung-Ho
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.185-190
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    • 2001
  • Alginate, a well-known biopolymer, is universally applied for immobilization of microbial cells. Biosorption characteristics of lead by waste biomass of immobilized A. niger beads, used in fermentation industries to produce citric acid, were studied. The immobilized A. niger beads, prepared via capillary extrusion method using calcium chloride, were applied in the removal of lead. Pb uptake was the highest in A. niger beads cells grown for 3 days with medium producing citric acid (12% sucrose, 0.5% $NH_4NO_3$, 0.1% $KH_2PO_4$, and 0.025% $MgSO_4$). Lead uptake by the immobilized A. niger beads and free A. niger mycellia beads increased sharply with time. However, while uptake by the immobilized A. niger beads continued to increase slowly, that by free A. niger mycellia beads stopped after 30 min. The optimum pH and temperature of lead uptake were found to be 6 and $35^{\circ}C$, respectively. The maximum uptake of lead was achieved with $50{\sim}100$ beads and 50 ml lead solution in a 250-ml Erlenmeyer flask, while, at over 100 beads, uptake of the lead decreased. The order of biosorption capacity for heavy metals was Pb>Cu>Cd. Pb uptake capacity of the immobilized A. niger beads treated with 0.1 M $CaCI_2$, 0.1 M NaOH, and 0.1 M KOH decreased compared to the untreated beads. On testing the desorption of Pb from the immobilized A. niger beads, re-uptake of Pb was found possible after desorption of the binding metal with 0.1 M HCI.

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Trichloroethylene (TCE) Removal Capacity of Synthesized Calcium Sulfoaluminate Minerals in Hydrated Cement-based Materials (합성 Calcium Sulfoaluminate계 시멘트 수화물의 Trichloroethylene (TCE) 제거능)

  • Ha, Min-Gyu;Ghorpade, Praveen A.;Kim, Jeong-Joo;Park, Joo-Yang
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.33 no.4
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    • pp.1463-1469
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    • 2013
  • Portland cement used as binding material in combination of ferrous iron for reductive dechlorination of chlorinated organics is already widely studied topic by several researchers. However there is no clear evidence about the component solely responsible in cement for trichloroethylene (TCE) dechlorination. Many researchers suspect that the ettringite, monosulfate phases associated with hydration of cement are responsible active agents for TCE dechlorination. This study deals with synthesizing different pure crystalline minerals like ettringite and monosulfate phases of cement hydration and check individual phase's TCE dechlorinating capacity in combination with ferrous iron. The results indicated that the synthesized minerals showed no reduction capacity for TCE. The findings in the present study is significant as it shows that ettringite and monosulfate phases which were suspected minerals by previous researchers for TCE dechlorination are not reactive. Hence it is suspected that some other mineral or mineral form in cement phase could be responsible for TCE degradation.

Solid-Phase Refolding of Poly-Lysine fusion Protein of hEGF and Angiogenin (Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘)

  • Park, Sang-Joong;Ryu, Kang;Suh, Chang-Woo;Chai, Young-Gyu;Kwon, Oh-Byung;Park, Seung-Kook;Lee, Eun-Kyu
    • KSBB Journal
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    • v.17 no.2
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    • pp.153-157
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.

Ultrastructure and Metallothionein Expression in Rat Liver Treated with Di-(2-ethylhexyl)phthalate (Di-(2-ethylhexyl)phthalate에 의한 흰쥐 간세포 미세구조와 metallothionein 발현에 미치는 영향)

  • Kim, Da-Ham;Moon, Seung-Hoon;Lee, Mi-Young;Lee, Jong-Hwa;Park, Young-Hyun;Shin, Kil-Sang;Kim, Wan-Jong
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.289-296
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    • 2007
  • Di-(ethyhexyl) phthalate (DEHP), commonly used as a plasticizer for manufacturing flexible vinyl products, has been the topic of extensive research, especially concerning endocrine disrupting properties. Metallothionein (MT) is a low molecular weight (6,000$\sim$7,000 Da), cysteine-rich (22$\sim$23%), metal-binding protein and is known to be induced by extrinsic factors such as chemical agents and stresses. Some of the known function of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Nonetheless, the definitive physiological function of MT are still unknown. This study was carried out to investigate the effects of DEHP on the ultrastructural changes and the expression of MT of the rat liver. The rats were orally intubated with either corn oil (experimental control) or 0.5 mg, 1.5 mg and 4.5 mg DEHP kg$^{-1}$ day$^{-1}$ in 0.5 mL of corn oil for 15 days before sacrificing and sampling. DEHP induced mild ultrastrctural changes of some cell organelles such as rough endoplasmic reticulum, mitochondria, lysosomes and peroxisomes in the rat liver treated with DEHP. In the respect of immunogold labelling and Western blotting, MT expression of the liver tissue was up-regulated by DEHP. In conclusion, DEHP has effects on the ultrastructures and hepatic function for MT expression in rat.

Process Optimization of PECVD SiO2 Thin Film Using SiH4/O2 Gas Mixture

  • Ha, Tae-Min;Son, Seung-Nam;Lee, Jun-Yong;Hong, Sang-Jeen
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.434-435
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    • 2012
  • Plasma enhanced chemical vapor deposition (PECVD) silicon dioxide thin films have many applications in semiconductor manufacturing such as inter-level dielectric and gate dielectric metal oxide semiconductor field effect transistors (MOSFETs). Fundamental chemical reaction for the formation of SiO2 includes SiH4 and O2, but mixture of SiH4 and N2O is preferable because of lower hydrogen concentration in the deposited film [1]. It is also known that binding energy of N-N is higher than that of N-O, so the particle generation by molecular reaction can be reduced by reducing reactive nitrogen during the deposition process. However, nitrous oxide (N2O) gives rise to nitric oxide (NO) on reaction with oxygen atoms, which in turn reacts with ozone. NO became a greenhouse gas which is naturally occurred regulating of stratospheric ozone. In fact, it takes global warming effect about 300 times higher than carbon dioxide (CO2). Industries regard that N2O is inevitable for their device fabrication; however, it is worthwhile to develop a marginable nitrous oxide free process for university lab classes considering educational and environmental purpose. In this paper, we developed environmental friendly and material cost efficient SiO2 deposition process by substituting N2O with O2 targeting university hands-on laboratory course. Experiment was performed by two level statistical design of experiment (DOE) with three process parameters including RF power, susceptor temperature, and oxygen gas flow. Responses of interests to optimize the process were deposition rate, film uniformity, surface roughness, and electrical dielectric property. We observed some power like particle formation on wafer in some experiment, and we postulate that the thermal and electrical energy to dissociate gas molecule was relatively lower than other runs. However, we were able to find a marginable process region with less than 3% uniformity requirement in our process optimization goal. Surface roughness measured by atomic force microscopy (AFM) presented some evidence of the agglomeration of silane related particles, and the result was still satisfactory for the purpose of this research. This newly developed SiO2 deposition process is currently under verification with repeated experimental run on 4 inches wafer, and it will be adopted to Semiconductor Material and Process course offered in the Department of Electronic Engineering at Myongji University from spring semester in 2012.

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pH, Alkaline Earth Metal Ion Effects and Miscibility with Hexadecanol on the Monolayer of Palmitic Acid at the Air-Water Interface (기-액 계면에서 Palmitic Acid 단분자막에 대한 pH, 알칼리토금속 이온의 영향 및 Hexadecanol 분자와의 섞임성)

  • Jong-Jae Chung;Byung-Il Seo;Hai-Won Lee;Min-Young Ju
    • Journal of the Korean Chemical Society
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    • v.37 no.3
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    • pp.294-301
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    • 1993
  • ${\pi}$-A isotherms of the Palmitic acid(PA) with increasing pH shifted to the low area/molecule due to the dissociation of PA at the air-water interface. More condensation of PA monolayers occurred by the addition of Mg$^{2+}$, Ca$^{2+}$ and Ba$^{2+}$ ion in subphase. This condensing effect was increased with increasing the concentration of these ions. Due to the interaction with each ion, PA were formed Mg, Ca, Ba-Palmitate complex. The binding structure between alkaline earth ion and carboxylate ligand in PA has been identified by IR spectrometry. The order of condensing effect of alkaline earth ions at pH 8 was Ca$^{2+}$ > Ba$^{2+}$+ > Mg$^{2+}$. The condensing effect except for Mg$^{2+}$ decreased with increasing atomic number. Whereas, the condensing effect in pure water system decreased with decreasing atomic number in the sequence: Ba$^{2+}$ > Ca$^{2+}$ > Mg$^{2+}$. The miscibility of binary system of PA and hexadecanol in monolayer showed that the miscibility was good for the pure water system. But, in the buffered pH 8 system, bad miscibility was found.

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Probiotic Properties of Lactic Acid Bacteria isolated from Feces and Kimchi (베트남인 분변 및 김치로부터 분리된 유산균의 프로바이오틱스 기능성 연구)

  • Shin, Hyun Su;Yoo, Sung Ho;Jang, Jin Ah;Won, Ji Young;Kim, Cheol Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.4
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    • pp.255-261
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    • 2017
  • The purpose of this study was to investigate the probiotic properties and antioxidant capacity of lactic acid bacteria isolated from Vietnamese feces and the Korean traditional food kimchi. Six isolated strains were identified as Lactobacillus sp. by 16S rRNA sequencing. All strains showed good resistance to low pH (1.5, 2.0, and 3.0) and 0.3% oxgall bile acids. Culture filtrates from the six strains showed various antioxidant effects, including DPPH, ABTS, reducing power, and metal chelating ($Fe^{2+}$) activities. Two of the six Lactobacillus strains showed potential probiotic activity. Heat resistance and adhesion assays were conducted by mixing the selected strains, Lactobacillus acidophilus V4, Lactobacillus plantarum V7, and Lactobacillus paracasei DK121 isolated from kimchi. The results showed that the heat resistance of these strains was similar to that of a commercial strain, L. plantarum LP. In addition, a mucin attachment assay using the mixture of selected strains (V4, V7, and DK121) showed high binding activity to the mucous layer. In conclusion, a mixture of V4, V7, and DK121 shows promising probiotic activity and may be useful for the development of health-related products.

Controlling the Location of Thermally Stable Au Nanoparticles with Tailored Surface Property within Block Copolymer Templates (열적으로 안정한 금나노입자를 이용한 블록공중합체 내에서의 입자위치 조절)

  • Kim, Se-Yong;Yoo, Mi-Sang;Jung, Se-Ra;Paek, Kwan-Yeul;Kim, Bum-Joon J.;Bang, Joona
    • Polymer(Korea)
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    • v.35 no.3
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    • pp.238-243
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    • 2011
  • Organic/inorganic hybrid materials have a lot of interest in various areas due to their fascinating properties. To control the location and dispersion of inorganic nanoparticles within polymer matrix. thiol-terminated polymeric ligands have been widely used to tune the surface property of nanoparticles. However, the specific binding between the thiol functional group and metal is unstable with increasing temperature. To archive the thermally-stable Au nanoparticles, we previously synthesized various UV-crosslinkable polymeric ligands, which have different compositions of polar, UV-crosslinkable azide unit comparing to non-polar 스티렌 units. After crosslinking the Au nanoparticles, it was found that the nanoparticles had superb stability at high temperature (above $180^{\circ}C$). In this work, we used thermally-stable Au nanoparticles to control the location within the polymer matrix. By changing the amount of polar azide units in the polymeric ligands, we could precisely control the location of nanoparticles from one domain to the interface of block copolymer templates.

Effect of Cu-resistant Pseudomonas on growth and expression of stress-related genes of tomato plant under Cu stress (구리-오염 토양에서 토마토 식물의 생장과 스트레스-관련 유전자 발현에 미치는 구리-내성 Pseudomonas의 영향)

  • Kim, Min-Ju;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.257-264
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    • 2017
  • Pseudomonas veronii MS1 and P. migulae MS2 have several mechanisms of copper resistance and plant growth promoting capability, and also can alleviate abiotic stress in plant by hydrolysis of a precursor of stress ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC deaminase. In 4-week pot test for tomato growth in soil contained 700 mg/kg Cu, inoculation of MS1 and MS2 significantly increased root and shoot lengths, wet weight and dry weight of tomato plants compared to those of uninoculated control. The inoculated tomato plants contained less amounts of proline that can protect plants from abiotic stress, and malondialdehyde, an oxidative stress marker than those of control. ACC synthase genes, ACS4 and ACS6, and ACC oxidase genes, ACO1 and ACO4, both involved in ethylene synthesis, were strongly expressed in Cu stressed tomato, whereas significantly reduced in tomato inoculated with MS1 and MS2. Also, a gene encoding a metal binding protein metallothionein, MT2 showed similar expression pattern with above genes. All these results indicated that these rhizobacteria could confer Cu resistance to tomato plant under Cu stress and allowed a lower level of Cu stress and growth promotion.