• Title/Summary/Keyword: TDP

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Cloning and Expression of Glucose-1-Phosphate Thymidylyltransferase Gene (schS6) from Streptomyces sp. SCC-2136

  • Han, Ji-Man;Kim, Su-Min;Lee, Hyo-Jung;Yoo, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.685-690
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    • 2007
  • The deoxysugar biosynthetic gene cluster of Sch 47554/Sch 47555 was cloned from Streptomyces sp. SCC-2136. One of the ORFs, schS6, appeared to encode glucose-1-phosphate thymidylyltransferase, which converts dTTP and glucose-1-phosphate to TDP-D-glucose and pyrophosphate. The dTDP-D-glucose is a key metabolite in prokaryotics as a precursor for a large number of modified deoxysugars, and these deoxysugars are a maj or part of various antibiotics, ranging from glycosides to macrolides. SchS6 was expressed in E. coli vector pSCHS6 and the expressed protein was purified to apparent homogeneity by ammonium sulfate precipitation and Ni-NTA affinity column chromatography. The specific activity of the purified enzyme increased 4.7-fold with 17.5% recovery. It migrated as a single band on SDS-PAGE with an apparent molecular mass of 56kDa. The purified protein showed glucose-1-phosphate thymidylyltransferase activity, catalyzing a reversible bimolecular group transfer reaction. In the forward reaction, the highest activity was obtained with combination of dTTP and ${\alpha}-D-glucose-1-phosphate$, and only 12% of that activity was obtained with the substrates $UTP/{\alpha}-D-glucose-1-phosphate$. In the opposite direction, the purified protein was highly specific for dTDP-D-glucose and pyrophosphate.

The Role of a Second Protein (Des VIII) in Glycosylation for the Biosynthesis of Hybrid Macrolide Antibiotics in Streptomyces venezuelae

  • HONG JAY SUNG JOONG;KIM WON SEOK;LEE SANG KIL;KOH HWA SOO;PARK HEE SUB;PARK SU JIN;KIM YOUN SANG;YOON YEO JOON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.640-645
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    • 2005
  • The function of the desVIII gene in the pikromycin producer Streptomyces venezuelae was characterized by gene deletion and complementation analysis. In addition to the DesVII glycosyltransferase, the desVIII gene that has previously been suggested to be required for the incorporation of endogenous deoxysugar, TDP-D-desosamine, into the aglycone of pikromycin is also required for the transfer of exogenous deoxysugars, TDP-D-quinovose and TDP-D-olivose.

Behavior of Nutrients in Runoff Water from a Small Rural Watershed (농촌 소유역 유출수에서의 영양염류의 거동)

  • Oh, Kwang-Young;Kim, Jin-Soo;Oh, Seung-Young
    • Journal of The Korean Society of Agricultural Engineers
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    • v.51 no.2
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    • pp.51-57
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    • 2009
  • The purpose of this study was to investigate characteristics of behavior of nutrients such as TN (Total nitrogen), TDN (Total dissolved nitrogen), TP (Total phosphorus) TDP (Total dissolved phosphorous) in runoff water from a nonpoint source dominated watershed ($6.67\;km^2$). Regular and intensive flow measurement and water sampling were taken during two years (February 2002 to January 2004) in the Ingyeong River, a tributary of the Han River. The mean concentrations of nutrients during rainy days were significantly (p < 0.05) higher than those during dry days. The mean TDN/TN ratio in rainy days (95%) is almost identical to that in dry days (96%), but mean TDP/TP ratio in rainy days (24%) significantly decreased compared with that in dry days (66%), suggesting that dominant form of TP is shifted from dissolved form to particulate form. Accordingly, the measures (.eg. filter strips, cover crops) to reduce soil erosion for fallow upland in the rainy season should be taken to control particulate phosphorous.

Crystallization and Preliminary X-ray diffraction Studies of Salmonella typhimurium Deoxy-Thymidine Diphosphate-D-Glucose-4, 6-Dehydratase (Salmonella typhimurium의 Deoxy-Thymidine Diphosphate-D-Glucose-4, 6-Dehydratase의 결정화와 X-선 회절에 관한 연구)

  • 최희욱;박교선
    • Korean Journal of Crystallography
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    • v.7 no.2
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    • pp.120-125
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    • 1996
  • Single crystals of deoxy-thymidine diphosphate-D-gluxose-4,6-dehydratase(abbreviated as dTDP-D-glucose dehydratase) from Escherichia coli Strain BL21 clone which harbors the gene of dTDP-D-glucose dehydratase in Salmonella typhimurium LT2 have been grown with and whithout substrates by sitting drop vapor diffusion at room temperature. The precipitating agent was 1.6 to 2.0 M Na, K phosphate buffer(pH 8.0). The crystals diffract to at least 2.5Å and belong to the hexagonal space group P61 with cell dimensions a=b=168.54Å, c=81.08Å. The asymmetric unit contains one dimer with a crystal volume per protein mass(VM) of 2.4Å3/Da and solvent content (Vsol) of 64% by volume.

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Pelvic Measurement Indicators to Produce the Gonad Protection of Female Infants (유아기 여아의 생식선 차폐체 제작을 위한 골반 지표 계측)

  • Kwak, Jong-hyeok;Park, Chan-hyeok;Moon, deog-hwan
    • Journal of the Korean Society of Radiology
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    • v.10 no.2
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    • pp.101-107
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    • 2016
  • It was performed in this study to provide basic information on production of accurate gonadal shield by measuring pelvic indicators of TDP SD ISP IAD 1CDP and 2CDP. when pelvis ap and hip ap examination was taken, there is no exact position of the reference point of the shield and anatomically more difficult to shield gonad in the case of female infants than male. Results analyzed by height in 70~80 and 110~120 were approximately 30mm 13mm 19mm 20mm 2mm and 7mm difference in TDP SD ISP IAD 1CDP and 2CDP respectively. This value was statistically significant (P<0.05). Results analyzed by age of 2~3 and 6~7 were different on SD ISP IAD and 2CDP by 17mm 10mm 12mm 16mm respectively. it was also statistically significant. However 1CDP was not statistically significant(P>0.05). the difference was nearly about 1mm. Analysis of results by weight of less than 10 and more than 20 showed difference on TDP SD ISP IAD 1CDP and 2CDP by 28mm 14mm 11mm 20mm 3mm and 8mm. it was statistically significant (P<0.05). From the above results, female infants pelvic indicator measurement can be used as reference value for shielding production and especially IAD can present a reference point on the shielding position. Pelvis indicators of female infant has a close correlation. In addition, as it is showen in the statistical difference analysis to have an accurate ovarian shield, gonad shield would be produced and used by height, age and weight.

지능형 금형공장 개발

  • Choe, Byeong-Gyu;Go, Gi-Hun;Kim, Bo-Hyeon
    • CDE review
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    • v.11 no.2
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    • pp.15-22
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    • 2005
  • Presented in the paper is an approach to developing an intelligent mold shop as a means to overcome the difficulties faced by mold-makers due to skill shortages and increased global competition. A machine shop where as much as of the human skills are replaced by a set of intelligent systems is called an intelligent machine shop, and an intelligent mold-making machine shop is called an intelligent mold shop(MS). By analyzing the contents of operator's skill. three intelligent S/W stations have been designed: Technical Data Processing(TDP) Station, Loading Schedule Station, and Real-time Monitoring Station. A detailed architecture of the TDP station is described, and measures of effectiveness of IMS are elaborated.

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Method for Cloning Biosynthetic Genes of Secondary Metabolites Including Deoxysugar from Actinomycetes

  • Sohng, Jae-Kyung;Oh, Tae-Jin;Kim, Chun-Gyu
    • BMB Reports
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    • v.31 no.5
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    • pp.475-483
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    • 1998
  • Many antibiotics contain partially deoxygenated sugar components that are usually essential for biological activity, affinity, structural stability, and solubility of antibiotics. Gene probes of the biosynthetic genes related with the deoxysugar were obtained from PCR. Primers were designed from the conserved peptide sequences of the known dTDP-D-glucose 4,6-dehydratases, which are the key step enzymes in the biosynthesis of deoxysugar. The primers were applied to amplify parts of dehydratase genes to 27 actinomycetes that produce the metabolites containing deoxysugar as structural constituents. About 180 and 340 bp DNA fragments from all of the actinomycetes were produced by PCR and analyzed by Southern blot and DNA sequencing. The PCR products were used as gene probes to clone the biosynthetic gene clusters for the antibiotic mithramycin, rubradirin, spectinomycin, and elaiophyrin. This method should allow for detecting of the biosynthetic gene clusters of a vast array of secondary metabolites isolated from actinomycetes because of the widespread existence of deoxysugar constituents in secondary metabolites.

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Studies on the possible role of thiamine in the central nervous system

  • Iwata, Heitaroh
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.51-55
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    • 1980
  • Thiamine, in the form of its diphosphate (TDP), is well known to act as a coenzyme, and during the early stage in the study of thiamine it had been believed that the symptoms of thiamine-deficiency were resulted secondarily from the disturbance of metabolic processes in which TDP participated as a coenzyme. However, the neurological symptoms in thiamine deficiency are now separated from the metabolic disturbances in thiamine deficiency. On the other hand, the specific involvement of phosphorylated thiamine in nerve conduction has been suggested by von Muralt, but nature of this involvement has not been elucidated at a molecular level. Recently the possible significance of thiamine triphosphate (TTP) in nervous tissue was suggested by the demonstration that TTP is not present in the brain of patients with subacute necrotizing encephalomyelitis, a fatal disease associated with an abnormality in thiamine metabolism. Furthermore, the studies using membrane fragments of rat brain strongly indicated that ion movement across the nerve membrane is associated with dephosphorylation of phosphorylated thiamine.

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Identification of High Affinity Non-Peptidic Small Molecule Inhibitors of MDM2-p53 Interactions through Structure-Based Virtual Screening Strategies

  • Bandaru, Srinivas;Ponnala, Deepika;Lakkaraju, Chandana;Bhukya, Chaitanya Kumar;Shaheen, Uzma;Nayarisseri, Anuraj
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3759-3765
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    • 2015
  • Background: Approaches in disruption of MDM2-p53 interactions have now emerged as an important therapeutic strategy in resurrecting wild type p53 functional status. The present study highlights virtual screening strategies in identification of high affinity small molecule non-peptidic inhibitors. Nutlin3A and RG7112 belonging to compound class of Cis-imidazoline, MI219 of Spiro-oxindole class and Benzodiazepine derived TDP 665759 served as query small molecules for similarity search with a threshold of 95%. The query molecules and the similar molecules corresponding to each query were docked at the transactivation binding cleft of MDM2 protein. Aided by MolDock algorithm, high affinity compound against MDM2 was retrieved. Patch Dock supervised Protein-Protein interactions were established between MDM2 and ligand (query and similar) bound and free states of p53. Compounds with PubCid 68870345, 77819398, 71132874, and 11952782 respectively structurally similar to Nutlin3A, RG7112, Mi219 and TDP 665759 demonstrated higher affinity to MDM2 in comparison to their parent compounds. Evident from the protein-protein interaction studies, all the similar compounds except for 77819398 (similar to RG 7112) showed appreciable inhibitory potential. Of particular relevance, compound 68870345 akin to Nutlin 3A had highest inhibitory potential that respectively showed 1.3, 1.2, 1.16 and 1.26 folds higher inhibitory potential than Nutilin 3A, MI 219, RG 7112 and TDP 1665759. Compound 68870345 was further mapped for structure based pharamacophoric features. In the study, we report Cis-imidazoline derivative compound; Pubcid: 68870345 to have highest inhibitory potential in blocking MDM2-p53 interactions hitherto discovered.