• Title/Summary/Keyword: Functional characterization

검색결과 793건 처리시간 0.028초

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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    • 제27권7호
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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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    • 제28권6호
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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.

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

  • 김명기;조석철
    • 융합정보논문지
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    • 제10권2호
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    • pp.102-108
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    • 2020
  • 지속적으로 감소하는 쌀 소비량을 해결하기 위해 쌀을 주원료로 하며 식사대용식은 물론 스낵으로도 제공될 수 있는 누룽지 제품의 다양성을 확보하고자 기능성을 보유한 7종의 천연 소재를 첨가하여 기호도와 상품성을 증가시킬 수 있는 누룽지를 제조하고 이화학적 특성을 분석하였다. 물 결합력은 흑임자를 첨가한 누룽지에서 264.13%로 가장 높게 나타났으며 경도는 갈색거저리를 첨가한 누룽지에서 0.36 N으로 가장 낮은 값을 보였다. 환원당 함량은 단호박을 첨가한 누룽지를 30분 침지하였을 때 1.47%로 가장 높게 나타났다. 다양한 소재를 첨가한 누룽지의 2,2-diphenyl-1-picryl-hydrazyl 유리라디칼 소거활성을 측정한 결과 단호박과 비트 첨가 누룽지에서 높은 소거능을 보였다. 관능평가 결과 전체적 기호도는 흑임자 첨가 누룽지에서 4.3으로 가장 높게 나타났다. 여러 소재가 첨가된 누룽지의 이화학적 특성 분석과 관능평가 결과 다양한 식품으로서의 적용이 가능한 것을 확인하였다.

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
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 춘계학술발표회
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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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폴리아크릴로니트릴계 활성나노탄소섬유의 기공특성이 이산화탄소 흡착에 미치는 영향 (Influence of the Pore Properties on Carbon Dioxide Adsorption of PAN-based Activated Carbon Nanofibers)

  • 이다영;조세호;김예솔;이영석
    • 폴리머
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    • 제37권5호
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    • pp.592-599
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    • 2013
  • Polyacrylonitrile(PAN) 고분자 용액으로부터 전기방사된 고분자 나노섬유를 다양한 농도의 KOH 용액을 이용하여 다공성 나노탄소섬유를 제조하였으며, 그에 따른 세공 구조 및 이산화탄소 흡착 특성을 평가하였다. PAN 용액으로부터 제조된 활성나노탄소섬유는 KOH 활성화 농도가 증가함에 따라 섬유 직경이 감소하였으며, 표면의 산소관능기가 증가하는 경향을 보였다. 또한 질소 흡착에 따른 세공특성을 분석한 결과 KOH 활성화 농도 증가에 따라 활성나노탄소섬유의 비표면적이 증가하고, 미세공은 4 M KOH로 활성화한 나노탄소섬유가 가장 많았으며, 중간세공은 8 M KOH로 활성화한 활성나노탄소섬유가 가장 많았다. 또한 0, $25^{\circ}C$에서 KOH 활성화제의 농도가 BET 및 XPS에서 나타난 것처럼 이산화탄소 흡착을 강화시키도록 세공 및 표면 특성에 영향을 주었다.

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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    • 제24권10호
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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.

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

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • 약학회지
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    • 제39권2호
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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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Sonocatalytic Degradation of Rhodamine B in the Presence of TiO2 Nanoparticles by Loading WO3

  • Meng, Ze-Da;Sarkar, Sourav;Zhu, Lei;Ullah, Kefayat;Ye, Shu;Oh, Won-Chun
    • 한국재료학회지
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    • 제24권1호
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    • pp.6-12
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    • 2014
  • In the present work, $WO_3$ and $WO_3-TiO_2$ were prepared by the chemical deposition method. Structural variations, surface state and elemental compositions were investigated for preparation of $WO_3-TiO_2$ sonocatalyst. X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX) and transmission electron microscopy (TEM) were employed for characterization of these new photocatalysts. A rhodamine B (Rh.B) solution under ultrasonic irradiation was used to determine the catalytic activity. Excellent catalytic degradation of an Rh.B solution was observed using the $WO_3-TiO_2$ composites under ultrasonic irradiation. Sonocatalytic degradation is a novel technology of treating wastewater. During the ultrasonic treatment of aqueous solutions sonoluminescence, cavitaties and "hot spot" occurred, leading to the dissociation of water molecules. In case of a $WO_3$ coupled system, a semiconductor coupled with two components has a beneficial role in improving charge separation and enhancing $TiO_2$ response to ultrasonic radiations. In case of the addition of $WO_3$ as new matter, the excited electrons from the $WO_3$ particles are quickly transferred to $TiO_2$ particle, as the conduction band of $WO_3$ is 0.74 eV which is -0.5 eV more than that of $TiO_2$. This transfer of charge should enhance the oxidation of the adsorbed organic substrate. The result shows that the photocatalytic performance of $TiO_2$ nanoparticles was improved by loading $WO_3$.

폴리프로필렌 복합소재의 아르곤 플라즈마 처리로 표면층 제거와 젖음성 향상 (Improvement of Wettability and Removal of Skin Layer on Ar-Plasma-Treated Polypropylene Blend Surface)

  • 원종일;이선용
    • 폴리머
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    • 제36권4호
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    • pp.461-469
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    • 2012
  • 아르곤(Ar) 플라즈마 처리된 폴리프로필렌 복합소재의 표면 개질 및 특성 변화를 X-선 광전자 분광 분석(XPS), 적외선 분광 분석(FTIR), 주사 전자 현미경 분석(SEM) 및 접촉각 측정 등을 이용하여 조사하였다. Ar 플라즈마 처리 시간의 증가는 폴리프로필렌 복합소재 표면의 젖음성, 극성 관능기를 갖는 산소 성분, 탈크 함량 및 표면조도의 증가를 초래하였다. 주사 전자 현미경 분석을 통한 자세한 관찰은 폴리프로필렌 성분으로 구성된 표면층(skin layer)이 존재함을 확인하였다. 폴리프로필렌과 고무 입자간의 점도차는 표면층의 생성을 촉진시켰다. 하지만 Ar 플라즈마 처리시간의 증가는 표면층의 두께를 감소시키는 것을 확인하였다. 사출성형 공정 동안, 표면층을 제거할 수 있는 추가적인 방법론에 대해서도 토의하였다. Ar 플라즈마 처리에 의한 표면 개질 및 모폴로지의 변화는 폴리프로필렌 복합소재 표면 상에 친수화 상태를 부여하고, 이에 따라 젖음성 향상을 유도하였다.

고분자 소재의 스크래치 거동의 정량적 평가 (Quantitative Evaluation of Scratch Behavior for Polymeric Materials)

  • 백기완;이성구;이재홍;최길영;원종일
    • 폴리머
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    • 제33권4호
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    • pp.273-283
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    • 2009
  • 고분자소재의 표면 특성 중의 하나인 스크래치 거동에 관한 최근의 연구 결과와 현황을 광범위하게 고찰하였다. 본 고찰에서는 고분자 소재의 스크래치 특성에 영향을 주는 실험인자들, 즉 소재재질, 온도, 작용하중, 실험속도, 표면처리, 스크래치 횟수, 고분자 구조 및 관능기, 가교정도 그리고 결정화도에 따른 스크래치 특성을 조사하였다. 또한, 고분자 소재의 스크래치 특성을 정량적으로 평가할 수 있는 접근 방법을 소개하였고, 현존하는 관련 국제시험법을 비교 조사하였다. 고분자 소재의 스크래치 거동을 정확히 평가할 수 있는 방법과 내스크래치성을 향상시킬 수 있는 방법에 관한 최근의 기술적 접근 방법에 대해 토의하였다.