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Responses of Bacteria to TNT: Cells′Survival, SDS-PAGE and 2-D Electrophoretic Analyses of Stress-Induced Proteins (TNT에 대한 세균의 반응기작: 생존율, 스트레스 유도단백질의 SDS-PAGE 및 2-D 전기영동 분석)

  • 오계헌;장효원;강형일;김승일
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.67-73
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    • 2002
  • The cellular responses of soil-borne bacterium, Pseudomonas sp. HK-6 to explosive 2,4,6-trinitrotoluene (TNT) were examined. Two stress shock proteins (SSPs), approximately 70-kDa DnaK and a 60-kDa GroEL were found in HK-6 cells in response to TNT. Analyses of SDS-PAGE and Western blot using anti-DnaK and GroEL revealed that SSPs were induced in HK-6 cells exposed to 0.5 M of TNT far 6-12 hrs. The maximum induction of proteins was achieved at 8-hr incubation point after HK-6 cells'exposure to TNT. Similar SSPs were found to be induced in HK-6 cells by heat shock (shift of temperature, from $30^{\circ}C$ to $42^{\circ}C$) or cold shock (shift of temperature,$30^{\circ}C$ to $4^{\circ}C$).2D-PAGE of soluble protein tractions from the culture of Pseudomonas sp. HX-6 exposed to TNT demonstrated that approximately 450 spots were observed on the silver stained gels ranging from pH 3 to pH 10. Among them, 12 spots significantly induced and expressed in response to TNT were selected and analyzed. Approximately 60-kDa protein, which was assumed highly expressed on the gel, was used for amino acid sequencing. N-terminal microsequencing with in-gel digestion showed that N-terminal sequence of the TNT-induced protein, <$^1XXAKDVKFGDSARKKML^17$, shared extensive similarity with $^1XXAKDVKFGDSARKKML^17$, N-terminal sequence of (P48216) GroEL of Pseudomonas putida.

Refractory Textile Wastewater Treatment Using Cell-Immobilized Polyethylene glycol Media (PEG 포괄고정화담체를 이용한 난분해성 염색폐수 처리)

  • Han, Duk-Gyu;Cho, Young-Jin;Bae, Woo-Keun;Hwang, Byung-Ho;Lee, Yong-Woo
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.3
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    • pp.345-350
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    • 2006
  • This study investigated the removal of recalcitrant organics in dyeing wastewater using a fluidized bed reactor(FBR) that contained cell-immobilized pellets. The pellets were manufactured and condensing the gel phase by mixing PEG-polymer and cells to form micro-porous PEG-polymer pellets whose size were ${\Phi}\;4mm{\times}H\;4mm$ on average. An industrial activated sludge without any pre-adaptation was used for the cell immobilization because it gave an equivalent removal efficiency to a pre-adapted sludges. The feed was obtained from an effluent of a biological treatment plant, which contained $SCOD_{Cr}$ of 330 mg/L and $SBOD_5$ of 20 mg/L. The $SCOD_{Cr}$ removal efficiency was over 45% and the effluent $COD_{Mn}$ concentration was less than 100 mg/L at HRTs from 6 to 24 hrs. The optimum HRT in the FBR was determined as 12 hrs considering the removal efficiency and cost. When a raw wastewater containing 768 mg/L of $COD_{Cr}$ was fed to the FBR, the effluent $COD_{Cr}$ concentration increased only slightly, giving a 70% of $COD_{Cr}$ removal or a 97% of $BCOD_5$ removal. This indicated that the FBR had an excellent capability of biodegradable organics removal also. In conclusion, the FBR could be applied to textile wastewater treatment in place of an activated sludge process.

Characterizations of Water-dispersed Biocellulose Nanofibers on the Skin Surface (피부 표면에서의 수분산 바이오셀룰로오스 미세섬유의 특성에 대한 연구)

  • Jun, Seung-Hyun;Kim, Seo Yeon;Park, Sun Gyoo;Lee, Cheon Koo;Lee, Seol-Hoon;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.89-94
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    • 2018
  • In this study, water-dispersed biocellulose nanofibers (TC) were prepared via an oxidation reaction using 2,2,6,6-tetramethyl-1-piperidine-N-oxy radical (TEMPO) as a catalyst. The TC retained their unique structure in water as well as in emulsion. TC adhered to the skin surface while maintaining nanofibrous structures, providing inherent functions of biocellulose, such as high tensile strength and high water-holding capacity. When gelatin gels as model skin were coated with TC, the hardness representing the elasticity was increased by 20% compared to untreated gelatin gel because TC could tightly hold the gelatin structure. When porcine skin was treated with TC and TC-contained O/W emulsion, the initial water contact angles of TC were lower than other materials, and dramatically decreased over time as water penetrated the fibrous structure of the TC film. Characterization of TC on the skin surface offered insight into the function of nanofibers on the skin, which is important for their applications with respect to fiber-cosmetics.

The Complete Amino Acid Sequence of Newborn Dog Prochymosin (강아지 프로카이모신의 전 아미노산 서열)

  • Yoon, Joo-Ok;Kim, Hyun-Ku
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.3
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    • pp.289-300
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    • 1997
  • Newborn dog chymosin was extracted from the stomachs of dogs of 2 weeks of age, and was purified by ion exchange chromatography. Half of the sequence was determined by amino acid sequencing and the complete sequence was deduced from a cloned chymosin cDNA Results showed that the zymogen showed 79% sequence identity with calf prochymosin and 54% identity with porcine pepsinogen A The size of the propart and location of the residue which becomes the amino-terminus in the active enzyme was the same in the prochymosins. The maximum general proteolytic activity at pH 3.2 of newborn dog chymosin was 3-4% of that of porcine pepsin A at pH 2, whereas the milk clotting activity relative to the general proteolytic activity of newborn dog chymosin was much higher than that of calf chymosin. Agar gel electrophoresis at pH 5.2 of stomach extracts of individual dogs showed the existence of two predominant genetic variants of zymogen and enzyme. The two variants could not be distinguished by amino acid composition or amino-terminal sequencing, and no differences in the enzymatic properties of the genetic variants were observed. It was concluded that of the residues that participate in the substrate binding, calf and newborn dog chymosin differ in the following positions (porcine pepsin numbering, subsites in parentheses) : Ser 12 Thr(S$_4$), Leu 30 Val(S$_1$/S$_3$), His 74 Gln(S'$_2$), Val 111 Ile(S$_1$/S$_3$), Lys 220 Met(S$_4$). With regard to the low general proteolytic activity of newborn dog chymosin, the substitution Asp303 Val relative to calf chymosin may contribute to an explanation of this.

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Exchange-coupling Interaction and Magnetic Properties of BaFe12O19/Ni0.5Zn0.5Fe2O4 Nanocomposite Ferrite (BaFe12O19/Ni0.5Zn0.5Fe2O4 나노복합체 Ferrite의 Exchange-coupling 상호 작용과 자기 특성)

  • Oh, Young-Woo
    • Journal of the Korean Magnetics Society
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    • v.24 no.3
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    • pp.81-85
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    • 2014
  • Nano-sized Ba-ferrite, Ni-Zn ferrite and $BaFe_{12}O_{19}/Ni_{0.5}Zn_{0.5}Fe_2O_4$ nanocomposite ferrite were prepared by sol-gel combustion method. Nanocomposite was calcined at temperature range of $600{\sim}900^{\circ}C$ for 1 h. According to the diffraction patterns, hard/soft nanocomposite was indicated to the coexistence of the magnetoplumbite structural $BaFe_{12}O_{19}$ and spinel $Ni_{0.5}Zn_{0.5}Fe_2O_4$ and agree with the standard data (JCPDS 10-0325). The particle size of nanocomposite turn out to be less than 90 nm. The nanocomposite ferrite shows a single-phase magnetization behavior, implying that the hard magnetic phase and soft magnetic phase were well exchange-coupled. The specific saturation magnetization ($M_s$) of the nanocomposite is located between hard ($BaFe_{12}O_{19}$) and soft ferrite ($Ni_{0.5}Zn_{0.5}Fe_2O_4$). The remanence (Mr) of nanocomposite ferrite is much higher than that for the individual $BaFe_{12}O_{19}$ and $Ni_{0.5}Zn_{0.5}Fe_2O_4$ ferrite. $(BH)_{max}$ is increased, generally.

Synthesis and Characterization of Temperature and pH Sensitive Graft Copolymers Based on Pluronic (Pluronic을 기초로 한 온도와 pH에 민감한 그래프트 공중합체의 합성과 특성)

  • Oh, Yeon-Jeong;Lee, Gi-Baek;Park, Sung-Young
    • Polymer(Korea)
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    • v.36 no.2
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    • pp.223-228
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    • 2012
  • Temperature and pH sensitive graft copolymers [Pluronic-$g$-poly(NIPAAm-$co$-MMA), Polymer A] and [Pluronic-$g$-poly( NIPAAm-$co$-MAA), Polymer C] were synthesized by macro radical graft polymerization with $N$-isopropylacrylamide (NIPAAM)/$N,N$-diethylaminoethylmethacrylate (DEAEMA) and $N$-isopropylacrylamide (NIPAAm)/methacrylic acid (MAA) based on Pluronic, respectively. The chemical structure and molecular weight of the graft copolymers was characterized by $^1H$ NMR and gel permeation chromatography. The aqueous solution properties of graft copolymers were measured using a UV-visible spectrophotometer, contact angle and dynamic light scattering equipment with different temperature and pH conditions. The obtained graft copolymers showed a very sensitive phase transition in response to temperature and pH in aqueous media which suggested that the amine group of DEAEMA segment and carboxylic group of MAA had a great influence on the lower critical solution temperatures (LCST) in Polymer A and C, respectively. The graft copolymers can be utilized for drug delivery system and molecular switching applications where responses to temperature and pH changes are relevant.

In situ Microfluidic Method for the Generation of Monodisperse Double Emulsions (미세유체를 이용한 단분산성 이중 에멀젼 생성 방법)

  • Hwang, So-Ra;Choi, Chang-Hyung;Kim, Hui-Chan;Kim, In-Ho;Lee, Chang-Soo
    • Polymer(Korea)
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    • v.36 no.2
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    • pp.177-181
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    • 2012
  • This study presents the preparation of double emulsions in a poly(dimethylsiloxane) (PDMS)-based microfluidic device. To improve the wettability of hydrophilic continuous phase onto a hydrophobic PDMS microchannel, the surface was modified with 3-(trimethoxysilyl) propyl methacrylate (TPM) and then sequentially reacted with acrylic acid monomer solution, which produced selective covalent bonding between acrylic acids and methacrylate groups. For the proof of selective surface modification, tolonium chloride solution was used to identify the modified region and we confirmed that the approach was successfully performed. When water containing 0.5% w/w sodium dodecyl sulfate and 1% w/w Span80 with hexadecane were loaded into the selectively modified microfluidic channels, we can produce stable double emulsion. Based on the spreading coefficients, we predict the morphology of double emulsions. Our proposed method efficiently produces monodisperse double emulsions having 48.5 ${\mu}m$(CV:1.6%) core and 65.1 ${\mu}m$ (CV:1.6%) shell. Furthermore, the multiple emulsions having different numbers of core were easily prepared by simple control of flow rates.

Sustained Release of Proteins Using Small Intestinal Submucosa Modified PLGA Scaffold (SIS로 개질된 PLGA 담체에서의 단백질의 서방화)

  • Ko, Youn-Kyung;Choi, Myung-Kyu;Kim, Soon-Hee;Kim, Geun-Ah;Lee, Hai-Bang;Rhee, John-M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.3
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    • pp.199-205
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    • 2008
  • In this study, we fabricated poly (lactide-co-glycolide) (PLGA) scaffold modified with small intestinal submucosa (SIS) as a drug delivery matrix of bioactive molecules. SIS derived from the submucosa layer of porcine intestine has been widely used as biomaterial because of low immune response. PLGA scaffold was prepared by the method of solvent casting/salt leaching. Novel composite scaffolds of SIS/PLGA were manufactured by simple immersion method of PLGA scaffold in SIS solution under vacuum. SEM observation shows that PLGA and SIS/PLGA scaffolds have interconnective and open pores. Especially, SIS/PLGA scaffold showed that micro-sponge of SIS with interconnected pore structures were formed in the pores of PLGA scaffold. In order to assay release profile of proteins, we manufactured FITC conjugated BSA loaded PLGA and SIS/PLGA scaffold. And the release amount was identified by fluorescence intensity using the fluorescence spectrophotometer. The initial burst of BSA containing SIS/PLGA scaffolds was lower than that of PLGA scaffolds resulting in constant release. And release of BSA in SIS/PLGA scaffold was fast and incremental because of the increased content of BSA. In conclusion, we confirmed that penetrated SIS solution prevented the initial burst of BSA and PLGA modified with SIS scaffold is useful as protein carriers with controlled release pattern.

Purification and Characterization of ($Ca^{2+}$+$Mg^{2+}$)-ATPase of Sarcoplasmic Reticulum from Rat Skeletal Muscle (쥐 근소포체의 ($Ca^{2+}$+$Mg^{2+}$)-ATPase의 분리정제와 그 효소특성에 관하여)

  • Lee, Jong-Soon;Ha, Doo-Bong;Chung, Chin-Ha
    • The Korean Journal of Zoology
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    • v.28 no.1
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    • pp.31-43
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    • 1985
  • The $(Ca^{2+}+Mg^{2+})$-ATPase has been purified homogeneously from sarcoplasmic reticulum of rat skeletal muscle by sucrose density gradient centrifugation. The purified enzyme has a molecular weight of 115,000 as judged by polyacrylamide gel electrophoresis in the presence of sodium dedecyl sulfate, and therefore has the same size of the enzyme in rabbit and chick skeletal muscle. $Ca^{2+}, Mg^{2+}, Fe^{2+}, Co^{2+}, and Mn^{2+}$ at 50 $\\muM$ show stimulatory effect on the ATP-ase, while $Zn^{2+}, Cu^{2+}, and Hg^{2+}$ inhibit it at the same concentration. The ATPase activity is insensitive to antimalarial drugs such as quinine and quinacrine, but is sensitive to inhibition by p-hydroxymecurie benzoate and phenylmethylsulfonylfluoride. The enzyme has optimum pH of 6 to 7 and Km value for ATP is estimated to be 98 $\\muM$. Thus, a number of biochemical properties of this enzyme appear to be different from those of the enzyme that have been isolated from rabbit skeletal muscle. The $(Ca^{2+}+Mg^{2+})$-ATPase appears to be selectively degraded in microsomal fraction. The activity of metalloendoprotease is evident in the microsomal preparation when assayed by radioactively labeled protein substrate, such as $^{3}H-casein and $^{125}I$-insulin. However, it is presently unclear whether the metalloendoprotease is responsible for the degradation of the $(Ca^{2+}+Mg^{2+})$-ATPase.

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Wear Resistance of Crosslinked Ultra-high Molecular Weight Polyethylene (가교된 초고분자량 폴리에틸렌의 내마모성)

  • Im, Chae-Ik;Lee, Gwi-Jong;Jo, Jae-Yeong;Choe, Jae-Bong;Choe, Gwi-Won
    • Journal of Biomedical Engineering Research
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    • v.20 no.1
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    • pp.99-106
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    • 1999
  • Ultra-high molecular weight polyethylene (UHMWPE) was crosslinked in the melt state to enhance wear resistance, Dicumyl peroxide (DCP) and triallyl cyanurate (TAC) was used as a crosslinking agent and a promoter, respectively. With increasing amount of DCP and TAC used, gel content of crosslinked UHMWPE (XUMPE) increased, while the melting temperature, crystallizaiton temperature, crystallinity, and tensile properties decreased. The results of pin-on-disk wear test and ball-on-disk test with small applied load showed reduced wear volumes of XUMPE from that of the unmodified UHMWPE. As the wear mechanism effected in the experimental condition of this study was thought to be deformation rather than adhesion or fatigue, a new parameter, the ratio of maximum contact stress to yield stress, was proposed to correlate well with observed wear resistance. In ball-on-disk wear test with larger applied load, XUMPE showed higher wear volumes than that of the unmodified UHMWPE which were accompanied with increased friction coefficients and surface roughness of the wear tracks. When contact stress was well above yield stress, the failure of XUMPE, as well as deformation, was thought to be much accelerated.

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