• Title/Summary/Keyword: Molecular structures

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Glucosylation of Isoflavonoids in Engineered Escherichia coli

  • Pandey, Ramesh Prasad;Parajuli, Prakash;Koirala, Niranjan;Lee, Joo Ho;Park, Yong Il;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.37 no.2
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    • pp.172-177
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    • 2014
  • A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-${\alpha}$-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the $5^{th}$ hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-${\beta}$-D-glucoside, genistein 7-O-${\beta}$-D-glucoside (genistin), genistein 4',7-O-${\beta}$-D-diglucoside, biochanin A-7-O-${\beta}$-D-glucoside (sissotrin), daidzein 4'-O-${\beta}$-D-glucoside, daidzein 7-O-${\beta}$-D-glucoside (daidzin), daidzein 4', 7-O-${\beta}$-D-diglucoside, and formononetin 7-O-${\beta}$-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOF-ESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli $BL21(DE3)/{\Delta}pgi{\Delta}zwf{\Delta}ushA$ overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of $200{\mu}M$ of supplemented isoflavonoids, without any additional UDP-${\alpha}$-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.

Conformational Study of Human Serum Albumin in Pre-denaturation Temperatures by Differential Scanning Calorimetry, Circular Dichroism and UV Spectroscopy

  • Rezaei-Tavirani, Mostafa;Moghaddamnia, Seyed Hassan;Ranjbar, Bijan;Amani, Mojtaba;Marashi, Sayed-Amir
    • BMB Reports
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    • v.39 no.5
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    • pp.530-536
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    • 2006
  • Thermal conformational changes of human serum albumin (HSA) in phosphate buffer, 10 mM at pH = 7 are investigated using differential scanning calorimetric (DSC), circular dichroism (CD) and UV spectroscopic methods. The results indicate that temperature increment from $25^{\circ}C$ to $55^{\circ}C$ induces reversible conformational changes in the structure of HSA. Conformational change of HSA are shown to be a three-step process. Interestingly, melting temperature of the last domain is equal to the maximum value of fever in pathological conditions, i.e. $42^{\circ}C$. These conformational alterations are accompanied by a mild alteration of secondary structures. Study of HSA-SDS (sodium dodecyl sulphate) interaction at $45^{\circ}C$ and $35^{\circ}C$ reveals that SDS affects the HSA structure at least in three steps: the first two steps result in more stabilization and compactness of HSA structure, while the last one induces the unfolding of HSA. Since HSA has a more affinity for SDS at $45^{\circ}C$ compared to $35^{\circ}C$, It is suggested that the net negative charge of HSA is decreased in fever, which results in the decrease of HSA-associated cations and plasma osmolarity, and consequently, heat removal via the increase in urine volume.

PCR Identification and Phylogenetic Analysis of Trichomonas gallinae from Domestic Pigeons in Guangzhou, China

  • Qiu, Shen-Ben;Lv, Meng-Na;He, Xi;Weng, Ya-Biao;Zou, Shang-Shu;Wang, Xin-Qiu;Lin, Rui-Qing
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.333-336
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    • 2017
  • Avian trichomoniasis caused by Trichomonas gallinae is a serious protozoan disease worldwide. The domestic pigeon (Columba livia domestica) is the main host for T. gallinae and plays an important role in the spread of the disease. Based on the internal transcribed spacers of nuclear ribosomal DNA of this parasite, a pair of primers (TgF2/TgR2) was designed and used to develop a PCR assay for the diagnosis of T. gallinae infection in domestic pigeons. This approach allowed the identification of T. gallinae, and no amplicons were produced when using DNA from other common avian pathogens. The minimum amount of DNA detectable by the specific PCR assay developed in this study was 15 pg. Clinical samples from Guangzhou, China, were examined using this PCR assay and a standard microscopy method, and their molecular characteristics were determined by phylogenetic analysis. All of the T. gallinae-positive samples detected by microscopic examination were also detected as positive by the PCR assay. Most of the samples identified as negative by microscopic examination were detected as T. gallinae positive by the PCR assay and were confirmed by sequencing. The positive samples of T. gallinae collected from Guangzhou, China, were identified as T. gallinae genotype B by sequencing and phylogenetic analyses, providing relevant data for studying the ecology and population genetic structures of trichomonads and for the prevention and control of the diseases they cause.

High frequency plant proliferation via direct fronds regeneration of Korean endemic duckweed species (국내 자생 좀개구리밥 (Lemna paucicostata) 및 개구리밥(Spirodela polyrhiza)의 엽상체 증식을 통한 기내 식물체 대량 증식체계 확립)

  • Oh, Myung-Jin;Park, Jong-Mi;Ko, Suk-Min;Liu, Jang R.;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.157-162
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    • 2009
  • High frequency plant proliferation system via direct frond regeneration of endemic duckweed plants Lemna paucicostata and Spirodela polyrhiza was established. Fronds of L. paucicostata and S. polyrhiza were able to multiply half-strength MS basal medium without plant growth regulators. However, addition of BA at a range of 1 to 3 mg/L was more effective than high concentration of BA treatments for fronds proliferation. Also half-strength MS salts was suitable for the fronds proliferation. Increase of salts concentration had inhibitory effect on fronds proliferation. Also the frequency of callus formation from fronds of L. paucicostata was 3.3%, when they cultured onto 1/2 MS medium supplemented with 1 mg/L of BA. Similarly the frequency of callus formation from S. polyrhiza was very low. After subculture of white globular structures derived from fronds of L. paucicostata, numerous globular somatic embryos and calluses were developed onto the surface of fronds. However these somatic embryos did not fully develop into normal plants when transferred to 1/2 MS basal medium. Therefore direct frond regeneration system was more efficient for mass proliferation of L. paucicostata and S. polyrhiza. The plant regeneration system of L. paucicostata and S. polyrhiza established in this study, might be applied to mass proliferation and genetic transformation for molecular breeding.

Genetic Characteristics of 207 Microsatellite Markers in the Korean Population and in other Asian Populations

  • Choi, Su-Jin;Song, Hye-Kyung;Jeong, Jae-Hwan;Jeon, In-Ho;Yoon, Ho-Sung;Chung, Ki Wha;Won, Yong-Jin;Choi, Je-Yong;Kim, Un-Kyung
    • Molecules and Cells
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    • v.25 no.2
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    • pp.301-304
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    • 2008
  • Microsatellites, short tandem repeats, are useful markers for genetic analysis because of their high frequency of occurrence over the genome, high information content due to variable repeat lengths, and ease of typing. To establish a panel of microsatellite markers useful for genetic studies of the Korean population, the allele frequencies and heterozygosities of 207 microsatellite markers in 119 unrelated Korean, Indian and Pakistani individuals were compared. The average heterozygosity of the Korean population was 0.71, similar to that of the Indian and Pakistani populations. More than 80% of the markers showed heterozygosity of over 0.6 and were valuable as genetic markers for genome-wide screening for disease susceptibility loci in these populations. To identify the allelic distributions of the multilocus genetic data from these microsatellite markers, the population structures were assessed by clustering. These markers supported, with the most probability, three clustering groups corresponding to the three geographical populations. When we assumed only two hypothetical clusters (K), the Korean population was separate from the others, suggesting a relatively deep divergence of the Korean population. The present 207 microsatellite markers appear to reflect the historical and geographical origins of the different populations as well as displaying a similar degree of variation to that seen in previously published genetic data. Thus, these markers will be useful as a reference for human genetic studies on Asians.

Nox4-Mediated Cell Signaling Regulates Differentiation and Survival of Neural Crest Stem Cells

  • Lee, Ji-Eun;Cho, Kyu Eun;Lee, Kyung Eun;Kim, Jaesang;Bae, Yun Soo
    • Molecules and Cells
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    • v.37 no.12
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    • pp.907-911
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    • 2014
  • The function of reactive oxygen species (ROS) as second messengers in cell differentiation has been demonstrated only for a limited number of cell types. Here, we used a well-established protocol for BMP2-induced neuronal differentiation of neural crest stem cells (NCSCs) to examine the function of BMP2-induced ROS during the process. We first show that BMP2 indeed induces ROS generation in NCSCs and that blocking ROS generation by pretreatment of cells with diphenyleneiodonium (DPI) as NADPH oxidase (Nox) inhibitor inhibits neuronal differentiation. Among the ROS-generating Nox isozymes, only Nox4 was expressed at a detectable level in NCSCs. Nox4 appears to be critical for survival of NCSCs at least in vitro as down-regulation by RNA interference led to apoptotic response from NCSCs. Interestingly, development of neural crest-derived peripheral neural structures in Nox4-/- mouse appears to be grossly normal, although Nox4-/- embryos were born at a sub-Mendelian ratio and showed delayed over-all development. Specifically, cranial and dorsal root ganglia, derived from NCSCs, were clearly present in Nox4-/- embryo at embryonic days (E) 9.5 and 10.5. These results suggest that Nox4-mediated ROS generation likely plays important role in fate determination and differentiation of NCSCs, but other Nox isozymes play redundant function during embryogenesis.

Physicochemical Properties of Chitin and Chitosan Prepared from Lobster Shrimp Shell (가시발새우 껍질에서 제조한 키틴.키토산의 물리화학적 특성)

  • Chung, Gea-Hwan;Kim, Bong-Sub;Hur, Jong-Wha;No, Hong-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.28 no.5
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    • pp.870-876
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    • 1996
  • The physicochemical properties of chitin and chitosan produced from lobster shrimp (Metanephrups thomosonii) shell were investigated. Lobster shrimp chitin contained 6.84% nitrogen, 0.57% fat and 0.32% ash, while chitosan contained 7.52% nitrogen, 0.13% fat and 0.33% ash. Degree of deacetylation and molecular weight of chitosan were 67.5% and $9.1{\times}105$, respectively. Yields of chitin from the shell portion and chitosan from the chitin were 15.7% and 75%, respectively. Chitin and chitosan contained 2.64 and 1.39mg/g of residual amino acids, respectively, with both the most predominant being lysine. Chemical structures of the lobster shrimp chitin and chitosan have been investigated by the IR and solid state $^{13}C-NMR$ spectra.

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Effects of Aloe-emodin and Emodin on Proliferation of the MKN45 Human Gastric Cancer Cell Line

  • Chihara, Takeshi;Shimpo, Kan;Beppu, Hidehiko;Yamamoto, Naoki;Kaneko, Takaaki;Wakamatsu, Kazumasa;Sonoda, Shigeru
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3887-3891
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    • 2015
  • Aloe-emodin (1, 8-dihydroxy-3-hydroxyl-methylanthraquinone; AE) and emodin (1,3,8-trihydroxy-6-methylanthraquinone; EM) are anthraquinone derivatives that have been detected in some medical plants and share similar anthraquinone structures. AE and EM have been shown to exhibit anticancer activities in various cancer cell lines; however, the inhibitory effects of these derivatives on the growth of cancer cells were previously reported to be different. Gastric cancer is the second most common cause of cancer cell death worldwide. In the present study, we examined the inhibitory effects of 0.05 mM AE and 0.05 mM EM on the proliferation of the MKN45 human gastric cancer cell line. The proliferation of MKN45 cells was significantly inhibited in AE- and EM-treated groups 24 h and 48 h after treatment. Furthermore, the inhibitory effects of EM were stronger than those of AE. The cell cycle of MKN45 cells were arrested in G0/G1 phase or G0/G1 and G2/M phases by AE and EM, respectively. However, an analysis of intracellular polyamine levels and DNA fragmentation revealed that the mechanisms underlying cell death following cell arrest induced by AE and EM differed.

The Crystal and Molecular Structure of Cholesteryl Hexanoate at Room and Low Tempreature (Cholesteryl Hexanoate의 실온 및 저온에서의 분자 및 결정구조)

  • Young Ja Park;B. M. Craven
    • Journal of the Korean Chemical Society
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    • v.25 no.3
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    • pp.131-139
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    • 1981
  • Cholesteryl hexanoate $(C_{33}H_{56}O_2)$, is monoclinic, space group $P2_1$, with a = 12.162(3), b = 9.314(3), c = 13.643(5) ${\AA}$, ${\beta}$ = $93. 55{\circ}(3)$ and two molecules per unit cell. The atomic coordinates from cholesteryl octanoate were used in an initial trial structure using X-ray intensities(Mo $K{\alpha}$ radiation) measured by a diffractometer at room temperature and $-75{\circ}C$. Structure refinement by block-diagonal least squares gave R = 0.129 and 0.105 for room and low temperature experiments respectively. The molecules are arranged in monolayers with their long axes antiparallel and severely tilted. There is a close packing of cholesteryl groups within the monolayers. The crystal structures is very similar to those of cholesteryl octanoate and cholesteryl oleate.

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Crystal Structure and Tautomerism Study of the Mono-protonated Metformin Salt

  • Wei, Xiaodan;Fan, Yuhua;Bi, Caifeng;Yan, Xingchen;Zhang, Xia;Li, Xin
    • Bulletin of the Korean Chemical Society
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    • v.35 no.12
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    • pp.3495-3501
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    • 2014
  • A novel crystal, the mono-protonated metformin acetate (1), was obtained and characterized by elemental analysis, IR spectroscopy and X-ray crystallography. It was found that one of the imino group in the metformin cation was protonated along with the proton transfer from the secondary amino group to the other imino group. Its crystal structure was then compared with the previously reported diprotonated metformin oxalate (2). The difference between them is that the mono-protonated metformin cations can be linked by hydrogen bonding to form dimers while the diprotonated metformin cations cannot. Both of them are stabilized by intermolecular hydrogen bonds to assemble a 3-D supermolecular structure. The four potential tautomer of the mono-protonated metformin cation (tautomers 1a, 1b, 1c and 1d) were optimized and their single point energies were calculated by Density Functional Theory (DFT) B3LYP method based on the Polarized Continuum Model (PCM) in water, which shows that the most likely existed tautomer in human cells is the same in the crystal structure. Based on the optimized structure, their Wiberg bond orders, Natural Population Analysis (NPA) atomic charges, molecular electrostatic potential (MEP) maps were calculated to analyze their electronic structures, which were then compared with the corresponding values of the diprotonated metformin cation (cation 2) and the neutral metformin (compound 3). Finally, the possible tautomeric mechanism of the mono-protonated metformin cation was discussed based on the observed phenomena.