• Title/Summary/Keyword: Iron production

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Optimization of the Expression of the Ferritin Protein Gene in Pleurotus eryngii and Its Biological Activity (큰느타리버섯에서 석충 페리틴 단백질 유전자의 발현 최적화 및 생물학적 활성)

  • Woo, Yean Jeong;Oh, Si Yoon;Choi, Jang Won
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.359-371
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    • 2019
  • To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

Electricity Generation and De-contamination Effect for Characteristic Electrode Material in a Microbial Fuel Cell System Using Bay Sediment (MFC의 금속 및 탄소전극에 의한 전기생산 특성과 오염저감 효과)

  • Kwon, Sung-Hyun;Song, Hyung-Jin;Lee, Eun-Mi;Cho, Dae-Chul;Rhee, In-Hyoung
    • Journal of Environmental Science International
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    • v.19 no.8
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    • pp.951-960
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    • 2010
  • Sediment works as a resource for electric cells. This paper was designed in order to verify how sediment cells work with anodic material such as metal and carbon fiber. As known quite well, sediment under sea, rivers or streams provides a furbished environment for generating electrons via some electron transfer mechanism within specific microbial population or corrosive oxidation on the metal surfaces in the presence of oxygen or water molecules. We experimented with one type of sediment cell using different anodic material so as to attain prolonged, maximum electric power. Iron, Zinc, aluminum, copper, zinc/copper, and graphite felt were tested for anodes. Also, combined type of anodes-metal embedded in the graphite fiber matrix-was experimented for better performances. The results show that the combined type of anodes exhibited sustainable electricity production for ca. 600 h with max. $0.57\;W/m^2$ Al/Graphite. Meanwhile, graphite-only electrodes produced max. $0.11\;W/m^2$ along with quite stationary electric output, and for a zinc electrode, in which the electricity generated was not stable with time, therefore resulting in relatively sharp drop in that after 100 h or so, the maximum power density was $0.64\;W/m^2$. It was observed that the corrosive reaction rates in the metal electrodes might be varied, so that strength and stability in the electric performances(voltage and current density) could be affected by them. In addition to that, COD(chemical oxygen demand) of the sediment of the cell system was reduced by 17.5~36.7% in 600 h, which implied that the organic matter in the sediment would be partially converted into non-COD substances, that is, would suggest a way for decontamination of the aged, anaerobic sediment as well. The pH reduction for all electrodes could be a sign of organic acid production due to complicated chemical changes in the sediment.

Studies on the Production of Enzymes by Thermophilic Actinomycetes (PART II) Some Properties of $\alpha$-Amylase from Thermophilic Actinomycetes (고온성 방선균에 의한 순소생산에 관한 연구 (제2보) $\alpha$-Amylase의 효소학적 성질)

  • Yang, Han-Chul;Park, Yong-Jin;Cho, Hong-Yeon
    • Microbiology and Biotechnology Letters
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    • v.4 no.3
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    • pp.91-97
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    • 1976
  • During the course of studies on the production and utilization of thermostable ${\alpha}$-amylase from a thormophilic actinomycete species isolated from soil, partial characterization of the ${\alpha}$-amylase has been (arried out. The optimum pH for the dextrinogenic activity of the enzyme was found to be 6.5 and the maximum reaction rate was achieved at a temperature range of 55$^{\circ}$ to 65$^{\circ}C$. Calcium ion was recognized to have a slight effect in activating the enzyme, while heavy metal salts especially ferrous and cupric ions showed a remarkable inhibition effect. The enzyme was best protected iron thermal denaturation at pH 8.0 with tris-HCI buffer;inactivation was rapid at higher or lower pH values. Furthermore, its thermal stability was greatly increased by calcium ion, particulary at the final concentration of 1${\times}$10$\^$-2/ mole in the reaction mixture. The Km value for the ${\alpha}$-amylase was calculated to be 2.17${\times}$10$\^$-4/g per $m\ell$ and the energy of activation for the dextrinogenic reaction to be 12,000${\pm}$580 ㎈ per mole.

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Effect of lactoferrin hydrolysates on inflammatory cytokine modulation in HEK-293, RBL-2H3, and HMC-1 cells

  • Son, Ji Yoon;Bae, Hyung Churl;Renchinkhand, Gereltuya;Nam, Myoung Soo;Kim, Woan-sub
    • Korean Journal of Agricultural Science
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    • v.47 no.1
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    • pp.83-93
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    • 2020
  • Lactoferrin (LF) is an iron-binding glycoprotein that is present in colostrum, milk, and other body secretions. The objective of this study was to investigate the effects of lactoferrin hydrolysates (LHs) on the production of immunomodulatory factors, including inflammatory related cytokines. The nuclear factor (NF)-κB reporter assay using human embryonic kidney 293 cells (HEK-293) revealed that NF-κB activity was significantly decreased by 1, 50, and 100 ㎍/mL of LH and the fractions above and below the 10 kDa LH. The mRNA expression of interferon (IFN)-γ in rat basophilic leukemia mast cells (RBL-2H3) treated with the fraction above the 10 kDa LH decreased in a dose-dependent manner, but the cells treated with LH and the fraction below the 10 kDa LH showed an increased expression of IFN-γ in a dose-dependent manner. The level of cyclooxygenase (COX)-2 expression decreased dose-dependently in RBL-2H3 cells treated with LH and the fraction above the 10 kDa LH, but the cells treated with the fraction below the 10 kDa LH showed an increased COX-2 expression in a dose-dependent manner. The mRNA expression of interleukin (IL)-4) was dose-dependently decreased by the fraction below the 10 kDa LH in human mast cells (HMC-1). The mRNA expressions of tumor necrosis factor (TNF)-α and IL-6 were significantly dose-dependently decreased by the fractions above and below the 10 kDa LH, but was dose-dependently increased by LH. The production of IL-4 was a little increased by the fraction above the 10 kDa LH compared to the positive control, but was decreased with LH and the fraction below the 10 kDa LH in HMC-1 cells. It was concluded that LF hydrolysates had an immunomodulating effect on anti-, pro-inflammatory and anti-allergic reactions.

Characteristics of Silica Coated ${\gamma}-Fe_{2}O_{3}$ with Heat-treatment (열처리에 따른 실리카 피착 ${\gamma}-Fe_{2}O_{3}$의 특성)

  • Lee, J.Y.;Byeon, T.B.;Kim, D.Y.;Lee, H.;Han, K.H.;Sohn, J.G.
    • Journal of the Korean Magnetics Society
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    • v.3 no.2
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    • pp.108-114
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    • 1993
  • This paper presents a study on the effects of silica coating in the production of ${\gamma}-Fe_{2}O_{3}$ powders suitable for magnetic recording media. Emphasis has been put on investigating the relationship between the powder characteristics and the effects of silica coating in the heat-treatment stage of ${\gamma}-Fe_{2}O_{3}$ production. After we prepared non-coated ${\gamma}-Fe_{2}O_{3}$ and silica coated ${\gamma}-Fe_{2}O_{3}$ with coating water glass on the surface of goethite and heattreatment process, we compared and investigated powder characteristics. As silica coated layer played a role of preventing the powders from overreduction to metal iron and rapid oxidation, silica coated ${\gamma}-Fe_{2}O_{3}$ showed superior magnetization value due to inhibiting t!1e adulteration of ${\alpha}-Fe_{2}O_{3}$ into the final product. When silica coated layer acted as a sintering restrainer, silica coated ${\gamma}-Fe_{2}O_{3}$ showed high coercivity and specific sur-face area due to good acicularity.

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Human Lung Insults due Air Pollutant -A Review for Priority Setting in the Research- (대기오염에 의한 폐장조직 손상 -연구방향의 설정을 위한 논의-)

  • 김건열;백도명
    • Journal of environmental and Sanitary engineering
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    • v.7 no.2
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    • pp.95-110
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    • 1992
  • Much progress has been made in understanding the subcellular events of the human lung injuries after acute exposure to environmental air pollutants. Host of those events represent oxidative damages mediated by reactive oxygen species such as superoxide, hydrogen peroxide, and the hydroxy, free radical. Recently, nitric oxide (NO) was found to be endogenously produced by endothelial cells and cells of the reticulo-endothelial system as endothelialderived relaxation factor (EDRF) which is a vasoactive and neurotransmitter substance. Together with superoxide, NO can form another strong oxidant, peroxonitrite. The relative importance of exogenous sources of $N0/N0_2$ and endogenous production of NO by the EDRF producing enzymes in the oxidative stresses to the heman lung has to be elucidated. The exact events leading to chronic irreversible damage are still yet to be known. From chronic exposure to oxidant gases, progressive epithelial and interstitial damages develop. Type I epithelial cells become thicker and cover a smaller average alveolar surface area while thee II cells proliferate instead. Under acute damages, the extent of loss of the alveolar epithelial cell lining, especially type II cells appears to be a good predictor of the ensuing irreversible damage to alveolar compartment. Interstitial matrix undergo remodeling during chronic exposure with increased collagen fibers and interstitial fibroblasts. However, Inany of these changes can be reversed after cessation of exposure. Among chronic lung injuries, genetic damages and repair responses received particular attention in view of the known increased lung cancer risks from exposure to several air pollutants. Heavy metals from foundry emission, automobile traffics, and total suspended particulate, especially polycystic aromatic hydrocarbons have been positively linked with the development of lung cancer. Asbestos in another air pollutant with known risk of lung cancer and mesothelioma, but asbestos fibers are nonauthentic in most bioassays. Studies using the electron spin resonance spin trapping method show that the presence of iron in asbestos accelerates the production of the hydroxy, radical in vitro. Interactions of these reactive oxygen species with particular cellular components and disruption of cell defense mechanisms still await further studies to elucidate the carcinogenic potential of asbestos fibers of different size and chemical composition. The distribution of inhaled pollutants and the magnitude of their eventual effects on the respiratory tract are determined by pollutant-independent physical factors such as anatomy of the respiratory tract and level and pattern of breathing, as well as by pollutant-specific phyco-chemical factors such as the reactivity, solubility, and diffusivity of the foreign gas in mucus, blood and tissue. Many of these individual factors determining dose can be quantified in vitro. However, mathematical models based on these factors should be validated for its integrity by using data from intact human lungs.

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Low-dose of organic trace minerals reduced fecal mineral excretion without compromising performance of laying hens

  • Qiu, Jialing;Lu, Xintao;Ma, Lianxiang;Hou, Chuanchuan;He, Junna;Liu, Bing;Yu, Dongyou;Lin, Gang;Xu, Jiming
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.4
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    • pp.588-596
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    • 2020
  • Objective: The objective of this study was to investigate the effects of low doses of organic trace minerals (iron, copper, manganese, and zinc) on productive performance, egg quality, yolk and tissue mineral retention, and fecal mineral excretion of laying hens during the late laying period. Methods: A total of 405 healthy hens (HY-Line White, 50-week-old) were randomly divided into 3 treatments, with 9 replicates per treatment and 15 birds per replicate. The dietary treatments included feeding a basal diet + inorganic trace minerals at commercial levels (CON), a basal diet + inorganic trace minerals at 1/3 commercial levels (ITM), and a basal diet + proteinated trace minerals at 1/3 commercial levels (TRT). The trial lasted for 56 days. Results: Compared to CON, ITM decreased (p<0.05) egg production, daily egg mass, albumen height, eggshell strength, yolk Fe concentration, serum alkaline phosphatase activity and total protein, and increased (p<0.05) egg loss and feed to egg ratio. Whereas with productive performance, egg quality, yolk mineral retention, and serum indices there were no differences (p>0.05) between CON and TRT. The concentrations of Fe and Mn in the tissue and tibia were changed notably in ITM relative to CON and TRT. Both ITM and TRT reduced (p<0.05) fecal mineral excretion compared to CON. Conclusion: These results indicate that dietary supplementation of low-dose organic trace minerals reduced fecal mineral excretion without negatively impacting hen performance and egg quality.

Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity (pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kang-Sun;Chung, Wee-Sup;Kim, Soo-Gwan;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.211-217
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    • 2006
  • Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

Synthesis of High-Quality Single-Walled Carbon Nanotube Fibers by Vertical CVD (수직 가열로를 이용한 고순도 단일벽 탄소나노튜브 섬유의 합성)

  • Kim, Tae-Min;Song, Woo-Seok;Kim, Yoo-Seok;Kim, Soo-Youn;Choi, Won-Chel;Park, Chong-Yun
    • Journal of the Korean Vacuum Society
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    • v.19 no.5
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    • pp.377-384
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    • 2010
  • Many routes have been developed for the synthesis of signle-walled carbon nanotubes (SWCNTs). We spun fibers of SWCNTs directly from vertical furnace using a liquid source of carbon and an iron-contained molecule. The solution was prepared by ethanol as a carbon source, in which ferrocene as a catalyst, thiophene were dissolved. It was then injected from the top of the furnace into hot zone with hydrogen as a carrier gas. We successfully synthesized high-quality SWCNTs by adjusting the various experimental conditions, such as concentration of ferrocene, solution injection rate, concentration of thiophene, and hydrogen flow rate. Measurement of Raman spectroscopy, scanning electron microscopy, and transmission electron microscopy were carried out to find the optimized conditions. The synthesized SWCNTs (1.16~1.64 nm) appeared a bundle structure and well-aligned parallel to the direction of furnace. These results also provide an simple way for high-quality SWCNTs mass production and fabricating direct spining SWCNTs fiber. It will allow one-step production of SWCNTs fiber with potentially excellent properties and wide-range applications.