• Title/Summary/Keyword: nad1

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Genetic and Morphologic Identification of Spirometra ranarum in Myanmar

  • Jeon, Hyeong-Kyu;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Sohn, Woon-Mok;Hong, Sung-Jong;Chai, Jong-Yil;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.275-280
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    • 2018
  • In the present study, we identified a Spirometra species of Myanmar origin (plerocercoid) by molecular analysis using mitochondrial cox1 and nad1 genes, as well as by morphological observations of an adult tapeworm. Spargana specimens were collected from a paddy-field in Taik Kyi Township Tarkwa Village, Yangon, Myanmar in December 2017. A total of 5 spargana were obtained from 20 frogs Hoplobatrachus rugulosus; syn: Rana rugulosa (Wiegmann, 1834) or R. tigrina (Steindachner, 1867). The plerocercoids were used for experimental infection of a dog. After 4 weeks of infection, an adult tapeworm was recovered from the intestine of the dog. Morphologically, the distinct features of Spirometra sp. (Myanmar origin) relative to S. erinaceieuropaei and S. decipiens include a uterine morphology comprising posterior uterine coils that larger than the terminal uterine ball and coiling of the uteri diagonally (swirling) rather than spirally. The cox1 sequences (1,566 bp) of the Myanmar-origin Spirometra species showed 97.9% similarity to a reference sequence of S. decipiens (GenBank no. KJ599679) and 90.5% similarity to a reference sequence of S. erinaceieuropaei (GenBank no. KJ599680). Phylogenetic tree topologies were identical and presented high confidence level of values for the 3 major branches of the 3 Spirometra species in cox1 and nad1 genes. These results indicated that Myanmar-origin Spirometra species coincided with those of S. ranarum and may be considered as a valid species.

Hansenula sp. MS-364의 생육과 Formate Dehydrogenase의 활성

  • 유병욱;권태종
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.403-407
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    • 1997
  • Medium components for maximum activity of NAD$^{+}$-dependent formate dehydrogenase (EC 1.2.1.2; FDH) were optimized with a methanol-assimilating yeast Hansenula sp. MS-364, preserved by our laboratory. The maximum activity of the enzyme was obtained when the strain was cultivated at 30$circ$C for 24 hours in a medium containing methanol 3%(v/v), yeast extract 0.8%(w/v), K$_{2}$HPO$_{4}$, 0.1%(w/v), KH$_{2}$PO$_{4}$ 0.1%(W/V), MgSO$_{4}$, 7H$_{2}$O 0.05%(w/v), and the pH of the culture broth was adjusted at 5.0.

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Isoforms of Glucose 6-Phosphate Dehydrogenase in Deinococcus radiophilus

  • Sung, Ji-Youn;Lee, Young-Nam
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.318-325
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    • 2007
  • Glucose 6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in Deinococcus radiophilus, an extraordinarily UV-resistant bacterium, was investigated to gain insight into its resistance as it was shown to be involved in a scavenging system of superoxide $(O_2^{-1})$ and peroxide $(O_2^{-2})$ generated by UV and oxidative stresses. D. radiophilus possesses two G6PDH isoforms: G6PDH-1 and G6PDH-2, both showing dual coenzyme specificity for NAD and NADP. Both enzymes were detected throughout the growth phase; however, the substantial increase in G6PDH-1 observed at stationary phase or as the results of external oxidative stress indicates that this enzyme is inducible under stressful environmental conditions. The G6PDH-1 and G6PDH-2 were purified 122- and 44-fold (using NADP as cofactor), respectively. The purified G6PDH-1 and G6PDH-2 had the specific activity of 2,890 and 1,033 U/mg protein (using NADP as cofactor) and 3,078 and 1,076 U/mg protein (using NAD as cofactor), respectively. The isoforms also evidenced distinct structures; G6PDH-1 was a tetramer of 35 kDa subunits, whereas G6PDH-2 was a dimer of 60kDa subunits. The pIs of G6PDH-1 and G6PDH-2 were 6.4 and 5.7, respectively. Both G6PDH-1 and G6PDH-2 were inhibited by both ATP and oleic acid, but G6PDH-1 was found to be more susceptible to oleic acid than G6PDH-2. The profound inhibition of both enzymes by ${\beta}-naphthoquinone-4-sulfonic$ acid suggests the involvement of lysine at their active sites. $Cu^{2+}$ was a potent inhibitor to G6PDH-2, but a lesser degree to G6PDH-1. Both G6PDH-1 and G6PDH-2 showed an optimum activity at pH 8.0 and $30^{\circ}C$.

The NAD(P)H: Quinine Oxidoreductase 1 (NQO1) Gene 609 C>T Polymorphism is Associated with Gastric Cancer Risk: Evidence from a Case-control Study and a Meta-analysis

  • Hu, Wei-Guo;Hu, Jia-Jia;Cai, Wei;Zheng, Min-Hua;Zang, Lu;Wang, Zheng-Ting;Zhu, Zheng-Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2363-2367
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    • 2014
  • The association between the NAD(P)H:quinone oxidoreductase 1 (NQO1) gene C609T polymorphism (rs1800566) and gastric cancer has been widely evaluated, but a definitive answer is so far lacking. We first conducted a case-control study to assess this association in a large Han Chinese population, and then performed a meta-analysis to further address this issue. Although our case-control association study indicated no significant difference in the genotype and allele distributions of C609T polymorphism between gastric cancer patients and controls, in the meta analysis involving 4,000 subjects, comparison of alleles 609T and 609C indicated a significantly increased risk (46%) for gastric cancer (95% confidence interval (95%CI) for odds ratio (OR)=1.20-1.79) in individuals with the T allele. The tendency was similar to the homozygote (OR=1.81, 95%CI: 1.16-2.84), dominant models (OR=1.41, 95%CI: 1.12-1.79), as well as recessive model (OR=1.58, 95%CI: 1.06-2.35). Stratified analysis by study design demonstrated stronger associations in population-based than in hospital-based studies. And ethnicity-based analysis demonstrated a significant association in Asians. We conclude that the NQO1 gene C609T polymorphism increases the risk for gastric cancer, especially in Asian populations.

Dissociation Constant of the Primary Amines and Quaternary Ammonium-methylorange Salts.

  • Kim, Bak-Kwang
    • YAKHAK HOEJI
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    • v.18 no.3
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    • pp.197-202
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    • 1974
  • The data pf dosspcoatopm cpmstamts for aliphatic amines and quaternary ammonium-methylorange salts are based on the electrosatic theory of conductance. Dissociation constants for primary amines nad quaternary ammonium-methylorange salts (1$\times$10$^{-5}$~1$\times$10$^{-3}$ M) in nitrobenzene solution or water solution was evaluated from the relation of the concentration and the electric conduction and the electric conductance at $25^{\circ}C$ Plots of ${\delta}C$ against reciprocal conductance were linear ; hence the center-to-center distance of this salt was 1.75 $\AA$ in nitrobenzene solution.

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Overexpression, Purification, and Biochemical Characterization of the Thermostable NAD-dependent Alcohol Dehydrogenase from Bacillus stearothermophilus

  • Shim, Eun-Jung;Jeon, Sang-Hoon;Kong, Kwang-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.738-744
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    • 2003
  • The gene ADH encoding NAD-dependent alcohol dehydrogenase from Bacillus stearothennophilus was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed fusion protein was purified simply by glutathione affinity chromatography. GST fusion protein was then cleaved by thrombin, while soluble enzyme was further purified by glutathione affinity chromatography. The recombinant enzyme had the same elctrophoretic mobility as the native enzyme from Bacillus stearothennophilus. The recombinant enzyme catalyzed the oxidation of a number of alcohols and exhibited high activities towards secondary alcohols. The $K_m\;and\;V_{max}$ values of the recombinant enzyme for ethanol were 5.11 mM and 61.35 U/mg, respectively. Pyridine and imidazole notably inhibited the enzymatic activity. The activity of the recombinant enzyme optimally proceeded at pH 9.0 and $70^{\circ}C$. The midpoint of the temperature-stability curve for the recombinant enzyme was approximately $68^{\circ}C$, and the enzyme was not completely inactivated even at $85^{\circ}C$. The recombinant enzyme showed a high resistance towards denaturing agents (0.05% SDS, 0.1 M urea). Therefore, due to its stability and relatively broad substrate specificity, the recombinant enzyme could be utilized in bio-industrial processes and biosensors.

Generation and characterization of calmodulin-DHFR sandwich fusion protein

  • Han, Chang Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.243-250
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    • 2008
  • A calmodulin-dihydrofolate reductase (DHFR) sandwich fusion protein was generated by insertion of calmodulin into the $\beta$-bulge region of DHFR to observe the effects of structurally constraining the calmodulin structure. The calcium binding properties of the sandwich protein were almost identical to calmodulin. Similar to calmodulin ($10.7 {\mu}M$), the sandwich protein bound four equivalents of calcium, with half saturation ($K_{0.5}$) observed at a [$Ca^{2+}$] of $8{\mu}M$. However, nicotinamide adenine dinucleotide (NAD) kinase activation property of the sandwich protein was lower than that of calmodulin. The sandwich protein activated NAD kinase, but to only half of the level obtained with calmodulin. The K 0.5 for both calmodulin and the sandwich protein were approximately the same (1-2 nM). Methylation analyses of the sandwich protein show that insertion of calmodulin into DHFR results in a large decrease in methylation. The $V_{max}$ observed with the sandwich protein (95 nmole/min/ml) was only 22% of the value observed with calmodulin (436 nmol/min/ml) in the presence of calcium. Addition of trimethoprim to the reaction significantly inhibited the observed methylation rate. Overall, the data suggest that the insertion of calmodulin into the DHFR structure has little effect on calcium binding by the individual lobes of calmodulin, but may constrain the lobes in a manner that results in altered interaction with the calmodulin-dependent proteins, and severely perturbed the methyltransferase recognition site.

Study on the Production of L-Latic Acid from Soluble Starch by Streptococcus sp. JEJ-6 (Streptococcus sp. JEJ-6에 의한 가용성 전분으로부터 L-Lactic Acid 생성에 관한 연구)

  • 전홍기;조영배;전은주;백형석
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.425-432
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    • 1998
  • The strain producing L-lactic acid from starch was isolated from kimchi. The isolated strain was identified as a homofermentative Streptococcus sp. through its morphological, cultural, biochemical characteristics, and named Streptococcus sp. JEJ-6. Lactic acids are of two types, one L-specific and the other D-specific form in a stereospecific form. Streptococcus sp. JEJ-6 produced selectively L-lactic acid from all of the tested carbon sources. The optimum conditions for the L-lactic acid production from the isolated microorganism were determined. For the maximum yield of L-lactic acid from Streptococcus sp. JEJ-6, the cell should be harvested at the early stationary phase, and the growth temperature, pH, and NaCl concentration should be 37$^{\circ}C$, pH 7.0 and 0.1%, respectively. 4% Soluble starch as substrate and organic nitrogen sources such as peptone and yeast extract should be used for the best yield. The optimum pH of the nicotinamide adenine dinucleotide(NAD)-dependent and NAD-independent lactate dehydrogenase(LDH) activities was pH 8.5 and pH 7.0, respectively.

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The conservation of a gilt-bronze Sarira Reliquary, Treasure No. 955 (보물 제955호 선암사 금동팔각원당형사리탑 보존처리)

  • Go, Hyeong-Sun;Yu, Jae-Eun
    • 보존과학연구
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    • s.24
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    • pp.215-227
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    • 2003
  • The gilt-bronze Sarira Reliquary was discovered when repairing three-stories east stone pagoda (Treasure No. 395) at Seungju-eup in Suncheon city in Jeollanam-do Province in August, 1986. Then it was appointed as Treasure in 1988. The Sarira Reliquary had been held in Seonamsa temple, but deterioration on the surface and corrosion had appeared affecting its surface detail. Consequently, the conservation treatment was carried out from November 2002 to March 2003.The corrosion and dirt on the surface of the Sarira Reliquary were cleaned with ethyl alcohol and Benzotriazole was applied to prevent further corrosion. Finally, NAD-10(Paraloid NAD-10), acrylic resin, was used to consolidate the structure. Moreover, after non-destructive analysis to confirm element of alloy, copper, gold, silver and mercury were discovered and this result tells us that it was plated with gold by amalgam. Fibers at the pedestal were examined under the microscope and identified as silk. The total height of this Sarira Reliquary is 6.0cm, the height of lotus pedestal and the roof is 2.7cm and 1.8cm, respectively. The roof and body are joined together, and the lotus pedestal can be separated, on which the octagonal reliquary is impaled. The pedestal consists of 3layers of petals and the surface is decorated with flower pattern. The reliquary is presumed to be created in the 14th century, and it becomes valuable historical material to reveal the secret of metal work in the late Goryeo Dynasty.

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Amperometric Determination of Urea Using Enzyme-Modified Carbon Paste Electrode

  • Yang, Jae-Kyeong;Ha, Kwang-Soo;Baek, Hyun-Sook;Lee, Shim-Sung;Seo, Moo-Lyong
    • Bulletin of the Korean Chemical Society
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    • v.25 no.10
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    • pp.1499-1502
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    • 2004
  • An amperometric biosensor based on carbon paste electrodes (CPEs) for the determination of urea was constructed by enzyme (urease/GL-DH)-modified method. Urea was hydrolyzed to ${NH_4}^+$ by catalyzing urease onto the enzyme-modified electrode surface in sample solution. In the presence of ${\alpha}$-ketoglutarate and reduced nicotinamide adenine dinucleotide(NADH), a liberated ${NH_4}^+$ produce to L-glutamate and $NAD^+$ by Lglutamate dehydrogenase (GL-DH). After the chemical reaction was proceeded, the electrochemical reaction was occurred that an excess of the NADH was oxidized to $NAD^+$. The oxidation current of NADH was monitored at +1.10 volt vs. Ag/AgCl. An optimum conditions of biosensor were investigated: The optimum pH range for catalyzed hydrolysis reaction of urea was pH 7.0-7.4. The linear response range and detection limit were $2.0\;{\times}\;10^{-5}{\sim}2.0\;{\times}\;10^{-4}M\;and\;5.0\;{\times}\;10^{-6}M$, respectively. Another physiological species did not interfere, except L-ascorbic acid.