• Title/Summary/Keyword: Molecular Separation

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Spatial protein expression of Panax ginseng by in-depth proteomic analysis for ginsenoside biosynthesis and transportation

  • Li, Xiaoying;Cheng, Xianhui;Liao, Baosheng;Xu, Jiang;Han, Xu;Zhang, Jinbo;Lin, Zhiwei;Hu, Lianghai
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.58-65
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    • 2021
  • Background: Panax ginseng, as one of the most widely used herbal medicines worldwide, has been studied comprehensively in terms of the chemical components and pharmacology. The proteins from ginseng are also of great importance for both nutrition value and the mechanism of secondary metabolites. However, the proteomic studies are less reported in the absence of the genome information. With the completion of ginseng genome sequencing, the proteome profiling has become available for the functional study of ginseng protein components. Methods: We optimized the protein extraction process systematically by using SDS-PAGE and one-dimensional liquid chromatography mass spectrometry. The extracted proteins were then analyzed by two-dimensional chromatography separation and cutting-edge mass spectrometry technique. Results: A total of 2,732 and 3,608 proteins were identified from ginseng root and cauline leaf, respectively, which was the largest data set reported so far. Only around 50% protein overlapped between the cauline leaf and root tissue parts because of the function assignment for plant growing. Further gene ontology and KEGG pathway revealed the distinguish difference between ginseng root and leaf, which accounts for the photosynthesis and metabolic process. With in-deep analysis of functional proteins related to ginsenoside synthesis, we interestingly found the cytochrome P450 and UDP-glycosyltransferase expression extensively in cauline leaf but not in the root, indicating that the post glucoside synthesis of ginsenosides might be carried out when growing and then transported to the root at withering. Conclusion: The systematically proteome analysis of Panax ginseng will provide us comprehensive understanding of ginsenoside synthesis and guidance for artificial cultivation.

Color-Tuning Mechanism of the Lit Form of Orange Carotenoid Protein

  • Man-Hyuk Han;Hee Wook Yang;Jungmin Yoon;Yvette Villafani;Ji-Young Song;Cheol Ho Pan;Keunwan Park;Youngmoon Cho;Ji-Joon Song;Seung Joong Kim;Youn-Il Park;Jiyong Park
    • Molecules and Cells
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    • v.46 no.8
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    • pp.513-525
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    • 2023
  • Orange carotenoid protein (OCP) of photosynthetic cyanobacteria binds to ketocarotenoids noncovalently and absorbs excess light to protect the host organism from light-induced oxidative damage. Herein, we found that mutating valine 40 in the α3 helix of Gloeocapsa sp. PCC 7513 (GlOCP1) resulted in blue- or red-shifts of 6-20 nm in the absorption maxima of the lit forms. We analyzed the origins of absorption maxima shifts by integrating X-ray crystallography, homology modeling, molecular dynamics simulations, and hybrid quantum mechanics/molecular mechanics calculations. Our analysis suggested that the single residue mutations alter the polar environment surrounding the bound canthaxanthin, thereby modulating the degree of charge transfer in the photoexcited state of the chromophore. Our integrated investigations reveal the mechanism of color adaptation specific to OCPs and suggest a design principle for color-specific photoswitches.

Preparation of Dextran Microparticles by Using the SAS Process (초임계 반용매 재결정 공정을 이용한 Dextran 입자의 제조)

  • Kang, Dong-Yuk;Min, Byoung-Jun;Rho, Seon-Gyun;Kang, Choon-Hyoung
    • Korean Chemical Engineering Research
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    • v.46 no.5
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    • pp.958-964
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    • 2008
  • In this work, micro-sized dextran particles, which have recently been focused as one of the candidate materials for the Drug Delivery System(DDS), were prepared by means of the Supercritical Antisolvent (SAS) process with $CO_2$. With dimethyl sulfoxide(DMSO) as the solvent, effects of the operating variables such as temperature (308.15~323.15 K), pressure(90~130 bar), solute concentration(10~20 mg/ml), and the molecular weight of the solute(Mw=37,500, 450,000) on the size and morphology of the resulting particles were thoroughly observed. The higher solute concentration led to the larger particles, however, the injection velocity of the solution and pressure did not show significant effects on the resulting particle size. With dextran of the lower molecular weight, the smallest particles were obtained at 313.15 K. On the other hand, the size of the particles from the high molecular weight dextran ranged between $0.1{\sim}0.5{\mu}m$ with an incremental effect of the temperature and pressure. For the solute concentration of 5 mg/ml, the lower molecular weight dextran did not form discrete particles while aggregation of the particles appeared when the solute concentration exceeded 15 mg/ml for the higher molecular weight dextran. It is believed that if the solute concentration is too low, the degree of the supersaturation in the recrystallization chamber would not be sufficient for initiation of the nucleation and growth mechanism. Instead, the spinodal decomposition mechanism leads to formation of the island-like phase separation which appears similar to aggregation of the discrete particles. This effect would be more pronounced for the smaller molecular weight polymer system due to the narrower phase-splitting region.

Gas Separation Study of PEBAX 3533 and PEG Blended Membranes (PEBAX 3533과 PEG의 혼합막에 대한 기체투과 연구)

  • Kim, Kwang Bae;Cho, Eun Hye;Cheong, Seong Ihl;Lee, Hyung Keun;Rhim, Ji Won
    • Membrane Journal
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    • v.23 no.2
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    • pp.144-150
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    • 2013
  • In order to increase the permeabilities of $N_2$, $O_2$, $CH_4$, $CO_2$, $SO_2$, Poly (ether block amides) (PEBAX) 3533 and its blended membranes with Poly (ethylene glycol) (PEG) of molecular weight 400 were prepared. The contents of PEG400 were 20%, 40%, and 50% and this membranes were characterized in terms of permeability for $N_2$, $O_2$, $CH_4$, $CO_2$, $SO_2$ gases and also diffusivity and solubility as well by using the time-lag gas separation apparatus. As expected, the permeabilities incerased as the contents of PEG400 increased. For the ideal selectivity, there is no big difference in values of between PEBAX 3533 and PEBAX/PEG400 membranes. The increase of permeabilities is due to the increases of solubilities of gases in question and this will be explained in more detail.

Predictive Thermodynamic Model for Gas Permeability of Gas Separation Membrane (기체 분리막의 투과 특성 예측 모델식 개발)

  • Kim, Jong Hwan;Hong, Sung Kyu;Park, Sang Jin
    • Korean Chemical Engineering Research
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    • v.45 no.6
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    • pp.619-626
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    • 2007
  • It is of special interest in our membrane separation technology due to its low energy consumption and cost, relatively simple equipment, low investment and operation cost, et al. Full scale utilization of such processes can be widely utilized to the various fields. Using the difference of permeability of gas molecules between the filter layers, it is able to separate effectually pure gases from the mixed gases. In this paper, the membranes of PDMS, ${\gamma}-radiated$ PDMS, PTFE, PTFE-X are chosen to develop the predictive model for the separation of pure gases such as oxygen, nitrogen, hydrogen, and other gases from mixed gases. By utilizing the thermodynamic gas properties($\sigma$, $\varepsilon/k$) and experimental data of gas transport characteristics for different polymer membranes, it is able to develop the predictive model equation under the influence of temperature, pressure and polymer characteristics. Predictive model developed in this research showed good agreement with experimental data of gas permeability characteristics for develop four different polymer membranes. The proposed model can also be extended to the general equation for predicting the separation of gases based on the properties of polymeric membranes.

LIQUID CRYSTALLINE POLYURETHANES. PHYSICOCHEMICAL CHARACTERISTICS AND PERVAPORATION PROPERTIES

  • A-Grabczyk, Aleksandra Wolibsk
    • Proceedings of the Membrane Society of Korea Conference
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    • 1996.04a
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    • pp.30-32
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    • 1996
  • Although the transport of small molecules through polymer membranes has been extensively studied for a long time, understanding of the transport mechanism is still far from satisfactory. This in turn makes difflcult the search for new membrane materials with the desired transport characteristics. Therefore it is of the utmost interest to study the correlations between a polymer's structure and morphology and its transport properties. Generally, polyurethanes serve as excellent polymer materials for such studies since their physical and chemical properties can be widely and systematically modified by varying the length, composition and chemical structure of the hard and soft segments. In this paper liquid crystalline polyurethanes are presented as new membrane materials for liquld separation and their transport properties with respect to molecular and supermolecular-structure are discussed.

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Improved Technologies to Produce Heterologous Proteins in Recombinant Escherichia coli. (재조합 대장균에서 외래단백질 발현을 위한 기술개발)

  • 박용철;권대혁;이대희;서진호
    • KSBB Journal
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    • v.16 no.1
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    • pp.1-10
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    • 2001
  • Escherichia coli has been used as an expression work horse for foreign genes. This article summarized recent development in genetic engineering techniques for overproduction of medical proteins and industrial enzymes. Special emphasis was placed upon research activities concerning folding and refolding of inclusion bodies at genetic and fermentation levels. Plasmid and mRNA stabilization, development of strong inducible promoters, modification of translational elements and reduction of rpoteolytic degradation were carried out to elevate an expression level of a target protein. Optimization of culture conditions, improvement of denaturation and renaturation steps and coexpression of molecular chaperones or foldase were accomplished to produce active proteins in soluble form. Fusion protein systems with selective separation and surface display technology were also performed in an effort to make the E. coli expression system more effective and versatile.

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Thermal denaturation analysis of protein

  • Miyazawa, Mitsuhiro
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1628-1628
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    • 2001
  • Near infrared (NIR) spectroscopy is a powerful technique for non-destructive analysis that can be obtained in a wide range of environments. Recently, NIR measurements have been utilized as probe for quantitative analysis in agricultural, industrial, and medical sciences. In addition, it is also possible to make practical application on NIR for molecular structural analysis. In this work, Fourier transform near infrared (FT-NIR) measurements were carried out to utilize extensively in the relative amounts of different secondary structures were employed, such as Iysozyme, concanavalin A, silk fibroin and so on. Several broad NIR bands due to the protein absorption were observed between 4000 and $5000\;^{-1}$. In order to obtain more structural information from these featureless bands, second derivative and Fourier-self-deconvolution procedures were performed. Significant band separation was observed near the feature at $4610\;^{-1}$ ,. Particularly the peak intensity at $4525\;^{-1}$ shows a characteristic change with thermal denaturation of fibroin. The structural information can be also obtained by mid-IR and CD spectral. Correlation of NIR spectra with protein structure is discussed.

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Comparison of Endonuclease-Sensitive Sites by T4 Endonuclease V and UvrABC Nuclease Treatments Followed by Formamide or Sodium Hydroxide Denaturation

  • Chang, Yung-Jin
    • BMB Reports
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    • v.31 no.4
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    • pp.405-408
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    • 1998
  • Endonuclease-sensitive sites detected by T4 endonuclease V or UvrABC nuclease treatments were compared in the dihydrofolate reductase gene of UV-irradiated Chinese hamster ovary B-11 cells. The number of endonuclease-sensitive sites detected by T4 endonuclease V treatment followed by NaOH denaturation was twice that of formamide denaturation. Repeated treatment of damaged genomic DNA with T4 endonuclease V resulted in no further increase in the number of endonuclease-sensitive sites detected. The numbers of endonuclease-sensitive sites detected by UvrABC nuclease using each denaturation condition were similar. Sequential treatment with the two endonucleases using formamide denaturation resulted in twice the number of endonuclease-sensitive sites detected by treatment of each nuclease alone. Due to a lack of AP endonuclease activity these results suggest the presence of T4 endonuclease V-sensitive sites which could be complemented by alkaline gel separation or by UvrABC nuclease treatment.

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Stereoselective Ligand Exchange Reaction of trans-dichlorocobalt(III) complex contained SS-epm and racemi propane-1,2-diamine (SS-epm이 배위된 trans-dichlorocobalt(III) 착물과 라세미 propane-1,2-diamine과의 입체선택적 리간드 치환반응)

  • Kim, Dong-Yeub
    • Journal of the Korean Society of Industry Convergence
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    • v.3 no.1
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    • pp.77-82
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    • 2000
  • The stereoselective ligand exchange reaction of trans-$[Co(SS-epm)Cl_2]^+$ and racemic propane-1,2-diamine (rac-pn) produces the complex that is identified as $[CO(N)_6]^{3+}$ type of $[Co(SS-epm)(pn)]^{3+}$ by absorption spectrum. It is conceivable that the reaction mechanism involves substitution and isomerization. The calculated and experimentally determined ratios of the complexed enantiomeric substrates at equilibrium were as follows: $[Co(SS-epm)(pn)]^{3+}$,calcd 32 % / 68 %, exptl 19 % / 81 % R-pn / S-pn. It has been shown that the employment of molecular mechanics calculations as a predictive tool may lead to the design of chiral complexes that may be applied to the separation of racemic mixtures of simple bidentate ligands.

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