• Title/Summary/Keyword: Synthesis

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Expression and Cloning of the pmmC Gene Encoding Phosphomannomutase in Sphingomonas chungbukensis DJ77 (Sphingomonas chungbukensis DJ77 균주에서 Phosphomannomutase를 암호화하는 pmmC 유전자의 클로닝과 발현)

  • Kim Mi-Hye;Choi Jung-Do;Shin Malshick;Kim Young-Chang
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.84-89
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    • 2005
  • Phosphomannomutase (PMM) is a key enzyme in prokaryotes and eukaryotes, which catalyzes the conversion of ${\alpha}$-D-mannose 6-phosphate to ${\alpha}$-D-mannose 1-phosphate. The latter is the substrate for the synthesis of GDP-mannose, which serves as the mannosyl donor for many metabolic pathways in the cells. We report here on the isolation of a gene from a genomic library of Sphingomonas chungbukensis DJ77, the pmmC gene encoding phosphomannomutase. The gene was cloned into E. coli expression vector, and the sequence was analyzed. The ribosomal binding site GGAAG lays 5 bp upstream of the ORF of 750 bp, which is initiated by ATG codon and terminated by TAG. The predicted sequence of the enzyme consists of 249 amino acids with a molecular mass of 27.4 kDa and showed $86.9\%$ similarity to that of eukaryotic phosphomannomutase after bioinformatical analyses with the conserved domain search of NCBI. The purified gene product revealed the activity of phosphomannomutase. In conclusion, we confirmed that pmmC gene encodes phosphomannomutase actually.

Inhibitory effect of Aralia elata ethanol extract against skin damage in UVB-exposed human keratinocytes and human dermal fibroblasts (두릅순 에탄올 추출물의 인간유래 피부각질형성세포와 피부섬유아세포에서의 자외선에 의한 광노화 억제효과)

  • Yang, Jiwon;Kwak, Chungshil
    • Journal of Nutrition and Health
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    • v.49 no.6
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    • pp.429-436
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    • 2016
  • Purpose: Solar ultraviolet (UV) radiation causes inflammation and matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging such as wrinkle formation, dryness, and sagging. Activation of MMP is influenced by various molecules such as reactive oxygen species (ROS), proinflammatory cytokines, and transient receptor potential vanilloid type (TRPV)-1, which are increased in UV-irradiated skin cells. Aralia elata (AE) ethanolic extract was reported to inhibit ROS generation caused by UVB-irradiation in keratinocytes. In this study, we investigated the photoprotective effect of AE ethanolic extract on UVB-irradiated human keratinocytes (HaCaT) and human dermal fibroblasts (HDF). Methods: AE was freeze-dried, extracted in 70% ethanol, and concentrated. Skin cells were treated with AE extract for 24 h and then exposed to UVB ($55mJ/cm^2$). After 48 h of incubation, proinflammatory cytokines, MMP-1, type-1 procollagen, and TRPV-1 levels were measured by ELISA or Western blotting. Results: Treatment with AE extract ($100{\mu}g/mL$) significantly inhibited UVB-induced IL-6, IL-8, and $PGE_2$ production in HaCaT by 25.6%, 5.3%, and 70.2%, respectively, and also inhibited elevation of MMP-1 and TRPV-1 caused by UVB irradiation by 20.0% and 41.9%, respectively (p < 0.05). In HDF, AE extract treatment significantly inhibited both elevation of MMP-1 and reduction of type-1 procollagen caused by UVB irradiation (p < 0.05). In addition, type-1 procollagen was elevated by AE extract treatment in normal HDFs (p < 0.05). Conclusion: AE 70% ethanol extract has photoprotective ability via reduction of proinflammatory mediators, TRPV-1 and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that AE extract might be a good natural material to protect against UVB-induced premature skin aging.

Studies on the synthesis of phenylmercuric 8-oxyquinolinate and its fungicidal effect (Phenylmercuric 8-oxyquinolinate의 합성(合成)과 그 살균효과(殺菌效果)에 대하여)

  • Shu, Y.T.;Son, C.Y.;Lee, S.H.
    • Applied Biological Chemistry
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    • v.6
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    • pp.37-43
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    • 1965
  • 8-Hydroxyquinoline, known to have the therapeutic effect to fusarium; and to diminish the amount of evaporation because of reducing the size of the stomata, and a new compound, phenylmercuric 8-oxyquinolinate, were synthesized. The fungicidal effect and diminishing effect of evaporation in phenylmercuric 8-oxyquinolinate were studied and the results are as follows. 1) 8-Hydroxyquinoline(m.p. $74{\sim}75^{\circ}C$, white needle crystalline) was synthesized by Skraup's method, 2) Phenylmercuric 8-oxyquinolinate(m.p. $159{\sim}160^{\circ}C$, yellowish brown needle crystalline) was synthesized by reacting phenylmercuricacetate to 8-hydroxyquinoline. 3) The orders of the fungicidal effects are; a) To Cochliobolus miyabeanus P.M.A.

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Ultrastructure of the Androgenic Gland of the Freshwater Prawn, Macrobrachium nipponense (징거미새우, Macrobrachium nipponense의 Androgenic Gland 미세구조)

  • 김대현;강정하;김대중;한창희
    • Development and Reproduction
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    • v.3 no.1
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    • pp.53-58
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    • 1999
  • The androgenic gland secretes a hormone, androgenic gland hormone, which is believed to act on the differentiation of the primary, secondary, and behavioral sex characteristics in most malacostracan crustaceans. This report presents the ultrastructural morphology of the androgenic gland in the freshwater prawn Macrobrachium nipponense. This gland, located in the coxopodite of the last pair of walking legs, was attached to the subterminal region of the sperm duct. The gland was composed of simple cellular strands, encased by a fibrous sheath. Microvilli were situated in the fibrous sheath, especially at the edge of each cellular strand. The androgenic gland cells had the large and round nucleus and rough endoplasmic reticulum arranged either in spirals or in concentric circles throughout the cytoplasm of the cell. They also had the well-developed Golgi complex and long mitochondria with flat and transverse cristae. The Golgi complex was similar to microvesicular cluster, but usually in the shape of typical dictyosomes, These features of androgenic gland cells coincides well with the protein/peptide secretion in their function. However, despite the apparent ultastructual equipment for protein/peptide secretion, no accumulation of materials secreted were noticed in the cytoplasm. Therefore it is strongly suggested that the transient transportation of the materials into the hemocoel has occurred just after synthesis.

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Development of Matrix for the Immobilization of High Level Radioactive Waste : Study on the Synthesis of Ce-pyrochlore (고준위 핵페기물의 고정화를 위한 메트릭스 개발 : Ce파이로클로어 합성 연구)

  • ;;;Yudintsev, S. V²
    • Economic and Environmental Geology
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    • v.35 no.2
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    • pp.97-102
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    • 2002
  • Ce-pyrochlore (CaCe $Ti_2 $O_7)was synthesized to study its properties and phase relations in CaO-Ce $O_2$-Ti $O_2$ system because Ce-pyrochlore was known as a promising material for the immobilization of radioactive actinide. The samples were prepared from the high purity starling materials under the pressure of 200~400 kg/$\textrm{cm}^2$ at room temperature, and annealed at 1000~ 150$0^{\circ}C$. The Synthesized samples were analysed and indentified with XRD and SEM/EDS methods. The optimal formation condition of Ce-pyrochlore was at 130$0^{\circ}C$ under $O_2$ atmosphere and the chemical composition of it wasCa$Ca_{1-x}Ti_{2-y}O_{7-x-2y}$(x=0.03-0.05, y=0.02~0.04) At temperature between 130$0^{\circ}C$ 140$0^{\circ}C$, Ce-pyrochlore underwent rapidly the incongruent decomposition to perovskite. Ce-perovskite, a partial solid solution between perovskite and loparite (C $e_{0.66}$Ti $O_3$), was observed as a major phase above 140$0^{\circ}C$.>.

The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: III. Comparative Molecular Field Analysis (CoMFA) on Binding Affinities between Ligands of 2-(Cyclohexyloxy) Tetrahydrofurane Derivatives and Porcine Odorant Binding Protein (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: III. 2-(Cyclohexyloxy) Tetrahydrofurane 유도체와 Porcine Odorant Binding Protein 사이의 결합 친화력에 관한 비교 분자장 분석)

  • Sung Nack-Do;Park Chang-Sik;Jung Hoon-Sung;Seong Min-Kyu
    • Reproductive and Developmental Biology
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    • v.30 no.1
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    • pp.13-19
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    • 2006
  • To search of new porcine pheromonal odorants for biostimulation control system technologies to improve reproductive efficiency in livestock species, the comparative molecular field analysis (CoMFA) for binding affinity constant $(p(Od)_{50})$ between porcine odorant binding protein (pOBP) and ligands of odorant 2-(cyclohexyloxy) tetrahydrofurane derivatives as substrate molecule was conducted and discussed. In the optimized CoMFA model AIV with chirality $(C_1'(R),\;C_2(S))$ in substrate molecule and atom based fit alignment (A) of odorants, the statistical results showed the best predictability of the binding affinities $(p(Od)_{50})$ based on the LOO cross-validated value $r^2_{cv}.\;(q^2=0.886)$ and non-cross-validated conventional coefficient $(r^2_{ncv}.=0.984)$. the binding affinity constants exhibited a good correlation with steric (40.8%), electrostatic (14.6%) and hydrophobic (44.6%) factors of the substrate molecules. from the analytical results of the contour maps, which may give us some valuable informations to the modification of odorants for effective binding affinity.

Effect of Yeast Addition in Rice Straw Silage Fermentation (볏짚 Silage 발효를 위한 효모의 첨가 효과)

  • 옥지운;이상민;이신자;임정화;강태원;정희영;문여황;이성실
    • Journal of Animal Science and Technology
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    • v.48 no.5
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    • pp.691-698
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    • 2006
  • Three species of the yeast Saccharomyces cerevisiae, Humicola grisea and Candida glabrata were assumed as microbial inoculants for fermentation of rice straw silage. Four types of silage innoculated with three yeasts including control (non-treatment) were opened on day 1, 3, 6, 9, 15 and 20 after ensiling, and analyzed for fermentation status (pH, crude protein, microbial counts) and the microbial population attached with silage texture using SEM (Scanning Electron Microscopy). The results obtained were summarized as fallow; The pH of silage juice was decreased to 4.3 after 6th day of fermentation in the treatments innoculated with yeast, but was not changed at the ranges of 5.47 to 5.67 in control. Crude protein concentration of silage was increased by 38~41% with yeast inoculation compared to control. From SEM observation, it could be confirmed that crude protein concentration of silage was increased by microbial growth and SCP synthesis. The yeast Saccharomyces cerevisiae and Candida glabrata could be used as useful fermenters of rice straw silage.

Effect of Whey Protein Isolate and Lactobacillus spp. Cell Extracts on Intracellular Antioxidative Activities in Human Prostate Epitherial Cells (유청단백질 및 Lactobacillus spp. 추출물이 전립선 세포 내 항산화 활성에 미치는 영향)

  • 변정열;윤영호
    • Journal of Animal Science and Technology
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    • v.48 no.5
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    • pp.719-726
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    • 2006
  • Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

The Amount of Telomeric DNA and Telomerase Activity on Cattle Cells (소의 생리적 특성에따름 세포내 텔로미어 함량과 텔로머레이스 활성도 분석)

  • Choi, Duk-Soon;Cho, Chang-Yeon;Sohn, Sea-Hwan
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.445-456
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    • 2008
  • Telomeres consist of TTAGGG tandem repeated DNA sequences with specific proteins and locate at chromosome ends. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Telomerase is a ribonucleoprotein which has a template for the synthesis of telomeric DNA. This study was carried out to analyze the amount of telomeric DNA and telomerase activity in cattle cells. Analysis of the quantity of telomere in lymphocytes was done at different ages, sex and among Korean cattle and Holstein breeds. The telomerase activity was also analyzed in liver, brain, heart, kidney, and testis tissues of fetal calf and of 18 month old cattle. The amount of telomeres in lymphocytes and other tissue cells was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) technique using a telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol assay (TRAP). The amount of telomeric DNA on the lymphocytes during the whole life span was decreased along with age. Quantity of telomeres in Korean cattle was significantly higher than that in Holstein breed. The amount of telomeric DNA in males was significantly higher than that in females. Telomerase activity was up-regulated in most bovine tissues during fetal stage, but was down-regulated in most tissues at mature 18 month age except the testis cells. This study indicates that the amount of telomeres and telomerase activity of cells can be used as an age marker or/and a physiological marker of cattle.

Procaryotic Expression of Porcine Acid-Labile Subunit of the 150-kDa Insulin-like Growth Factor Complex (미생물에서 돼지 150-kDa Insulin-Like Growth Factor Complex의 Acid-Labile Subunit 발현)

  • Lee, C. Young;Kang, Hye-Kyeong;Moon, Yang-Soo
    • Journal of Animal Science and Technology
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    • v.50 no.2
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    • pp.177-184
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    • 2008
  • Acid-labile subunit(ALS) is a 85-kDa glycosylated plasma protein which forms a 150-kDa ternary complex with 7.5-kDa insulin-like growth factor(IGF) and 40~45-kDa IGF-binding protein-3. In a previous study, the present authors prepared a porcine ALS(pALS) expression construct by inserting a pALS coding sequence into a plasmid vector following synthesis of the sequence by reverse transcription-polymerase chain reaction(RT-PCR). The expression construct, however, was subsequently found to have a mis-sense mutation at two bases of the pALS coding sequence which is presumed to have occurred through a PCR error. In the present study, the correct coding sequence was synthesized by the site-directed mutagenesis and inserted into the pET-28a(+) plasmid expression vector containing the His-tag sequence flanking the last codon of the insert DNA. After induction of the expression construct in E. coli BL21(DE3) cells, the resulting presumptive recombinant peptide was purified by the Ni-affinity chromatography. Upon SDS- PAGE, the affinity-purified peptide was resolved as a single band at a 66-kDa position which is consistent with the expected molecular mass of the presumptive recombinant pALS. Collectively, results indicate that a recombinant pALS peptide was successfully expressed and purified in the present study.