• Title/Summary/Keyword: Functional characterization

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Preparation and Characterization of Fluoroacrylate Copolymer by Emulsion Polymerization (유화 중합법에 의한 불소아크릴레이트 공중합체 제조)

  • 김동옥;김지현
    • Polymer(Korea)
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    • v.27 no.6
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    • pp.528-535
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    • 2003
  • The effects of surfactants, organic solvents, and functional monomers on the emulsion polymerization of perfluoroalkyleoylacryaltes and n-alkylacrylates were investigated. In particular, the dependence of the surface properties, contact angle and water repellency on the crystal melting temperature (T$\_$m/) of the fluorocopolymer and the variation of polymer latex particle sizes was investigated. Using WAXD experiments and synthesizing different types of fluorocopolymers which have fallowing fluoroacrylaytes [CH$_2$=CHCO$_2$CH,$_2$(CF$_2$CF$_2$) nH] (n = 4, 5 or 6), the relationship between the molecular packing structure of pendent side groups of fluorocopolymers and the surface properties was also investigated. We observed that the structure of primary carbon atoms of pendent side groups of fluorocopolymers plays key role in determining the surface properties.s.

Characterization of a Soil Metagenome-Derived Gene Encoding Wax Ester Synthase

  • Kim, Nam Hee;Park, Ji-Hye;Chung, Eunsook;So, Hyun-Ah;Lee, Myung Hwan;Kim, Jin-Cheol;Hwang, Eul Chul;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.248-254
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    • 2016
  • A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

Molecular Characterization of Cold-Inducible ${\beta}$-Galactosidase from Arthrobacter sp. ON14 Isolated from Antarctica

  • Xu, Ke;Tang, Xixiang;Gai, Yingbao;Mehmood, Muhammad Aamer;Xiao, Xiang;Wang, Fengping
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.236-242
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    • 2011
  • A psychrotrophic bacterium, Arthrobacter sp. ON14, isolated from Antarctica, was shown to exhibit a high ${\beta}$-galactosidase activity at a low temperature. A genomic library of ON14 was constructed and screened for ${\beta}$-galactosidase genes on functional plates containing 5-bromo-4-chloro-3-indolyl-${\beta}$-D-galactopyranoside (X-gal) as the substrate. Two different ${\beta}$-galactosidase genes, named as galA, galB, were found in ON14. Computational analyses of the genes revealed that the encoded protein GalA belongs to family 2 of glycosyl hydrolysases and is a cold-active protein, whereas GalB belongs to family 42 of glycosyl hydrolysases and is a mesophilic protein. Reverse transcription analyses revealed that the expression of galA is highly induced at a low temperature ($4^{\circ}C$ ) and repressed at a high temperature ($28^{\circ}C$ ) when lactose is used as the sole carbon source. Conversely, the expression of galB is inhibited at a low temperature and induced at a high temperature. The purified GalA showed its peak activity at $15^{\circ}C$ and pH 8. The mineral ions $Na^+$, $K^+$, $Mg^{2+}$, and $Mn^{2+}$ were identified as enzyme activators, whereas $Ca^{2+}$ had no influence on the enzyme activity. An enzyme stability assay revealed that the activity of GalA is significantly decreased when it is incubated at $45^{\circ}C$ for 2 h, and all its activity is lost when it is incubated at $50^{\circ}C$.

Characterization of a Glutamate Decarboxylase (GAD) from Enterococcus avium M5 Isolated from Jeotgal, a Korean Fermented Seafood

  • Lee, Kang Wook;Shim, Jae Min;Yao, Zhuang;Kim, Jeong A;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1216-1222
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    • 2017
  • To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

Functional Characterization of the C-Terminus of YhaV in the Escherichia coli PrlF-YhaV Toxin-Antitoxin System

  • Choi, Wonho;Yoon, Min-Ho;Park, Jung-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.987-996
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    • 2018
  • Bacterial programmed cell death is regulated by the toxin-antitoxin (TA) system. YhaV (toxin) and Pr1F (antitoxin) have been recently identified as a type II TA system in Escherichia coli. YhaV homologs have conserved active residues within the C-terminus, and to characterize the function of this region, we purified native YhaV protein (without denaturing) and constructed YhaV proteins of varying lengths. Here, we report a new low-temperature method of purifying native YhaV, which is notable given the existing challenges of purifying this highly toxic protein. The secondary structures and thermostability of the purified native protein were characterized and no significant structural destruction was observed, suggesting that the observed inhibition of cell growth in vivo was not the result of structural protein damage. However, it has been reported that excessive levels of protein expression may result in protein misfolding and changes in cell growth and mRNA stability. To exclude this possibility, we used an [$^{35}S$]-methionine prokaryotic cell-free protein synthesis system in vitro in the presence of purified YhaV, and two C-terminal truncated forms of this protein (YhaV-L and YhaV-S). Our results suggest that the YhaV C-terminal region is essential for mRNA interferase activity, and the W143 or H154 residues may play an analogous role to Y87 of RelE.

Analysis of Physicochemical Properties of Nurungji Added with Various Materials (다양한 소재가 첨가된 누룽지의 이화학 특성 분석)

  • Kim, Myong-Ki;Cho, Seok-Cheol
    • Journal of Convergence for Information Technology
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    • v.10 no.2
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    • pp.102-108
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    • 2020
  • Increasing the rice consumption, seven kinds of Nurungji which served as a home meal replacer and snack were developed with some functional materials were added, and their physiochemical characteristics were analyzed. The water binding capacity was the highest in Nurungji with black sesame seed (264.13%), and the hardness was the lowest as 0.36 N in Nurungji with Tenebrio molitor. The reducing sugar content was the highest in Nurungji with sweet pumpkin (1.47%) when being soaked in water for 30 minutes. As a result of measuring 2,2-diphenyl-1-picryl-hydrazyl free radical scavenging activity of the tested Nurungji preparations, Nurungji with sweet pumpkin and beet showed the high scavenging activity against free radicals. As a result of sensory evaluation, overall acceptability was the highest in Nurungji with black sesame seeds (4.3). Physicochemical characterization and sensory evaluation of Nurongji added with various materials confirmed that they could be applied to various food products.

Physicochemical Characterization of Extrudate Solid Formulation of Angelica gigas Nakai Prepared by Hot Melt Extrusion Process

  • Azad, Md Obyedul Kalam;Cho, Hyun Jong;Koo, Ja Seong;Park, Cheol Ho;Kang, Wie Soo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.72-72
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    • 2018
  • The root of Angelica gigas Nakai (AGN) is used as a traditional herbal medicine in Korea for the treatment of many diseases. However, a major challenge associated with the usage of the active compounds from AGN is their poor water solubility. Therefore, this work aimed to enhance the solubility of active compounds by a chemical (viz. surfactant) and physical (hot melt extrusion) crosslinking method (CPC). Infrared Fourier transform spectroscopy (FT-IR) revealed multiple peaks in extrudate solids representing new functional groups including carboxylic acid, alkynes and benzene derivatives. Differential scanning calorimetry (DSC) analysis of the extrudate showed lower glass transition temperature (Tg) and lower enthalpy (${\Delta}H$) (Tg: $43^{\circ}C$; ${\Delta}H$: <6 (J/g)) compared to the non-extrudate (Tg $68.5^{\circ}C$; ${\Delta}H$: 123.2) formulations. X-ray powder diffraction (XRD) analysis revealed amorphization of crystal materials in extrudate solid. In addition, nanonization, enhanced solubility and higher extraction of phenolic compounds were achieved in the extrudate solid. Among the different extrudates, acetic acid- and Span 80-mediated formulations showed superior extractions. We conclude that the CPC method successfully enhanced the production of amorphous nano dispersions from extrudate solid formulations.

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Influence of the Pore Properties on Carbon Dioxide Adsorption of PAN-based Activated Carbon Nanofibers (폴리아크릴로니트릴계 활성나노탄소섬유의 기공특성이 이산화탄소 흡착에 미치는 영향)

  • Lee, Dayoung;Cho, Seho;Kim, Yesol;Lee, Young-Seak
    • Polymer(Korea)
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    • v.37 no.5
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    • pp.592-599
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    • 2013
  • In this study, polyacrylonitrile (PAN)-based porous carbon nanofibers were prepared from PAN polymer solution by electrospinning and KOH activation with various concentrations, and the characterization of pore structures and carbon dioxide adsorption was investigated. Manufactured PAN-based activated carbon nanofibers tend to decrease diameter and increase surface oxygen functional groups depending on the increasing concentration of KOH solution. In addition, according to the results of nitrogen adsorption for pore properties analysis, it indicated increase of the specific surface area in conformity with increasing concentration of KOH solution. Micropore volume of treated activated carbon nanofibers (ANCF) by 4 M KOH was the largest compared with other samples and mesopore volume of treated ANCF by 8 M KOH was the largest volume, respectively. The concentration of KOH effects textural and surface properties, as represented by BET and XPS, which enhance carbon dioxide adsorption capacity at 0 and $25^{\circ}C$.

Structural and Functional Characterization of CRAMP-18 Derived from a Cathelicidin-Related Antimicrobial Peptide CRAMP

  • Park, Kyong-Soo;Shin, Song-Yub;Hahm, Kyung-Soo;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.24 no.10
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    • pp.1478-1484
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    • 2003
  • CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.

Functional Characterization and Regional Expression of Dopamine Transporter (도파민 수송체의 기능적 특성 및 발현에 관한 연구)

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.161-168
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    • 1995
  • Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

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