• Title/Summary/Keyword: hydrophobic acid

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Molecular Cloning and Characterization of a cis-Epoxysuccinate Hydrolase from Bordetella sp. BK-52

  • Pan, Hai Feng;Bao, Wen Na;Xie, Zhi Peng;Zhang, Jian Guo;Li, Yongquan
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.659-665
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    • 2010
  • A cis-epoxysuccinate hydrolase (CESH) from Bordetella sp. BK-52 was purified 51.4-fold with a yield of 27.1% using ammonium sulfate precipitation, ionic exchange, hydrophobic interaction, molecular sieve chromatography and an additional anion-exchange chromatography. The CESH was stable in a broad range of temperature (up to $50^{\circ}C$) and pH (4.0-10.0) with optima of $40^{\circ}C$ and pH 6.5, respectively. It could be partially inhibited by EDTA-$Na_2$, $Ag^+$, SDS, and DTT, and slightly enhanced by $Ba^{2+}$ and $Ca^{2+}$. The enzyme exhibited high stereospecificity in D(-)-tartaric acid (enantiomeric excess value higher than 99%) with $K_m$ and $V_max$ values of 18.67 mM and $94.34\;{\mu}M$/min/mg for disodium cis-epoxysuccinate, respectively. The Bordetella sp. BK-52 CESH gene, which contained 885 nucleotides (open reading frame) encoding 294 amino acids with a molecular mass of about 32 kDa, was successfully overexpressed in Escherichia coli using a T7/lac promoter vector and the enzyme activity was increased 42-times compared with the original strain. It may be an industrial biocatalyst for the preparation of D(-)-tartaric acid.

Controlled Release of Paclitaxel from Biodegradable Polymer Films for Drug-Eluting Stents (약물방출 스텐트용 생분해성 고분자 필름으로부터 파크리탁셀의 조절 방출)

  • Kim, Si-Eun;Lee, Bong-Soo;Kim, Jin-Hyang;Park, Kwi-Deok;Han, Dong-Keun
    • Polymer(Korea)
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    • v.34 no.2
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    • pp.172-177
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    • 2010
  • Although many researchers have studied the efficacy of paclitaxel (PTX) on many cells during the last two decades, little work has been reported on the importance of release kinetics inhibiting cell proliferation. The aim of this study is to examine the release behavior of the PTX on various biodegradable polymers such as poly(lactic-co-glycolic acid)(PLGA), poly-L-lactide (PLLA), and polycaprolactone (PCL) for drug-eluting stents (DES). The PTX from the fabricated films was released for 8 weeks and the degree of degradation of the films was observed by FE-SEM. Although the degradation time of PCL was the slowest, the PTX release rate was the fastest among them and followed by PLGA and PLLA with the equivalent PTX concentration. It suggests that hydrophobic drug such as PTX from polymer with low $T_g$ like PCL could be moved easily and released rapidly in body temperature.

Self-organized Pullulan/Deoxycholic Acid Nanogels: Physicochemical Characterization and Anti-cancer Drug-releasing Behavior

  • Na, Kun;Park, Kyong-Mi;Jo, Eun-Ae;Lee, Kwan-Shik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.262-267
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    • 2006
  • The objective of this study was to develop new self-organized nanogels as a means of drug delivery in patients with cancer. Pullulan (PUL) and deoxycholic acid (DOCA) were conjugated through an ester linkage between the hydroxyl group in PUL and the carboxyl group in DOCA. Three types of PUL/DOCA conjugates were obtained, differing in the number of DOCA substitutions (DS; 5, 8, or 11) per 100 PUL anhydroglucose units. The physicochemical properties of the resulting nanogels were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of DS 11 was the smallest (approx. 100 nm), and the size distribution was unimodal. To determine the organizing behavior of these conjugates, we calculated their critical aggregation concentrations (CACs) in a 0.01-M phosphate buffered saline solution. They were $10.5{\times}10^{-4}mg/mL,\;7.2{\times}10^{-4} mg/mL,\;and\;5.6{\times}10^{-4} mg/mL$ for DS 5, 8, and 11, respectively. This indicates that DOCA can serve as a hydrophobic moiety to create self-organized nanogels. To monitor the drug-releasing behavior of these nanogels, we loaded doxorubicin (DOX) onto the conjugates. The DOX-loading efficiency increased with the degree of DOCA substitution. The release rates of DOX from PUL/DOCA nanogels varied inversely with the DS. We concluded that the PUL/DOCA nanogel has some potential for use as an anticancer drug carrier because of its low CAC and satisfactory drug-loading capacity.

Preferential Peroxidase Activity of Prostaglandin Endoperoxide H Synthase for Lipid Peroxides

  • Yun, Seol-Ryung;Han, Su-Kyong;Song, In-Seok
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.94-94
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    • 2001
  • Prostaglandin endoperoxide H synthase (PGHS) catalyzes the committed step in prostaglandins and thromboxane A$_2$-- oxygenation of arachidonic acid to the hydroperoxy endoperoxide PGG$_2$, followed by reduction PGG$_2$to the alcohol PGH$_2$. The two reactions by PGHS -- cyclooxygenase and peroxidase -- occur at distinct but structurally and functionally interconnected sites. The peroxidase reaction occurs at a heme-containing active site located near the protein surface. The cyclooxygenase reaction occurs in a hydrophobic channel in the core of the enzyme. Initially a peroxide reacts with the heme group, yielding Compound I and an alcohol derived from the oxidizing peroxide. Compound I next undergoes an intramolecular reduction by a single electron traveling from Tyr385 along the peptide chain to the proximal heme ligand, His388, and finally to the heme group. Following the binding of arachidonic acid, Tyr385 tyrosyl radical initiates the cyclooxygenase reaction by abstracting the 13-pro(5) hydrogen atom to give an arachidonyl radical, which sequentially reacts with two molecules of oxygen to yield PGG$_2$. In order to characterize PGHS peroxidase active site, we examined various lipid peroxides with purified recombinant ovine PGHS proteins and determined the rate constants. The results have shown that twenty-carbon unsaturated fatty acid hydroperoxides have similar efficiency in peroxidation by PGHS, irrespective of either the location of hydroperoxy group or the number of double bonds. It was also confirmed by the subsequent study with PGHS peroxidase active site mutants.

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Cloning, Expression, and Nucleotide Sequencing of the Gene Encoding Glucose Permease of Phosphotransferase System from Brevibacterium ammoniagenes

  • Yoon, Ki-Hong;Yim, Hyouk;Jung, Kyung-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.214-221
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    • 1998
  • A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

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Industrial Synthesis for α-Sulfonation of Fatty Acid Methyl Ester (알파술폰화 고급지방산 메틸 에스테르의 공업적 합성)

  • Kim, Jin-Hyun;Choi, Seung-Ok;Yun, Young-Kyun;Nam, Ki-Dae
    • Applied Chemistry for Engineering
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    • v.9 no.4
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    • pp.595-602
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    • 1998
  • ${\alpha}$-sulfonated fatty acid methyl esters[$C_mH_{2m+1}CH(SO_3Na)COOC_nH_{2n+1}$], where hydrophobic group has carbon number of 12~18, were prepard by sulfonation of fatty acid methylester. The mole ratio of $SO_3$ to ester used was 1.3 and the reaction temperature was $70{\sim}90^{\circ}C$. The yield was found to be 97% by mixed gas reaction of inactive gas/gaseous $SO_3$. Studies on bleaching and neutralization processes in the pilot scale provided conditions applicable to industrial synthesis process.

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The Study on Synthesis and Application of Polymer Dispersion for Cement Modifier -The Waterproffing Effeet of Cement Mortar by Poly[DMA-co-DAMA] Emulsion- (시멘트 혼화용 폴리머 합성과 그 응용에 관한 연구 -Poly[DMA-co-DAMA] 에멀젼을 이용한 시멘트 모르타르의 방수성-)

  • Kim, Young-Geun;Herh, Dong-Seop;Park, Hong-Soo
    • Applied Chemistry for Engineering
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    • v.5 no.4
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    • pp.669-680
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    • 1994
  • DMA-co-DAMA were synthesized from 2-diethylaminoethyl metacrylate and dodecyl-metacrylate containing long chain hydrocarbon group with hydrophilic and hydrophobic radicals. To facilitate water emulsification,acrylic copolymer was cationized by acetic acid to produce acetated acrylic copolymer. The structures of the synthesized copolymer and acetated copolymers were confirmed by IR, NMR, and molecular weight was measured by GPC, and C. H. N elemental analysis. Acetated acrylic copolymers were perfectly emulsified in water and showed increased emulsion stability. Polymer dispersion for cement modifier(PDCM-PDD) was prepared by blending of the guaternized acrylic copolymer synthesized above sodium silicate sodium gluconate oleic acid and triethanol amine. The result with prepared polymer dispersion of cement modfier was examined, and it was found that excellent waterproffing effect; Water permeability ratio is 0.44 under the water pressure of $100g/cm^2$ and 0.55 under $3kg/cm^2$, and water absorption ratio is 0.36~0.47 and 1.02 compressive strength ratio at mixed ratio of water/PDCM-PDD is 45 times.

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Biosynthesis of Penicillins and Cephalosporins Antibiotics (페니실린과 세파로스포린계 항생제의 생합성)

  • 김경자;구양모
    • YAKHAK HOEJI
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    • v.27 no.3
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    • pp.185-205
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    • 1983
  • Penicillins and cephalosporins are biosynthesized from L-.alpha.-aminoadipic acid, L-cysteine and L-valine. A tripeptide, LLD-$\delta$-($\alpha$-aminoadipyl)cysteinylvaline(LLD-ACV) was isolated from fermentation broths of Cephalosporium acremonium as well as of Penicillium chrysogenum and it was proved that the LL-$\delta$-($\alpha$-aminoadipyl cysteine was formed first in mycelia, to which valine would be connected to give LLD-ACV. However, several points are still unsolved; first, what mechanism is involved in the configurational change from L-valine to D-valine, second, what kind of cyclization mechanism gives a $\betha$-lactam ring and a thiazolidine ring and third, what is the pathways for the ring expansion from penicillins to cephalosporins. At present, it seems clear that LLD-ACV is cyclized to give isopenicillin N, which is transformed to penicillin N and further to cepbalosporin C. Other hydrophobic penicillins, including benzyl penicillin and penicillin V, are formed from isopenicillin N by acyl-exchange reactions catalyzed by penicillin transferase, rather than by acylation reaction on 6-aminopenicillanic acid(6-APA), which was isolated from the fermentation broth of P. chrysogenum and which would be formed by hydrolysis of $\delta-(\alpha$-amincadipyl)amido moiety at the C-6 position in isopenicillin N or penicillin N by penicillin acylase. Acylation of 6-APA is catalyzed also by penicillin acylase, but the reaction is proved not to be involved in penicillin biosynthesis. Understanding the biosynthesis of penicillins and cephalsoporins would provide solutions to increase in fermentation yields of penicillins, especially of cephalosporins and a solution to biological production of 7-aminocepbalosporanic acid (7-ACA) which is of importance in pharmaceutical industry. Still regulation mechanisms in penicillin and cephalosporin biosynthesis are unveiled at all.

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Formation of the multiamellar vesicles of AHAsomes and effect of removal on the horny layer (AHAsomes의 multilamellar vesicles형성과 각질제거 효과)

  • 김인영;서봉석
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.21 no.2
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    • pp.1-21
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    • 1995
  • In this context, it should be mentioned that multilamellar vesicles can be prepared with the main compounds of the intercellular lipids, ceramides, cholesterol, cholesteryl ester, squalane, lecithin, wax ester by effect of the wetting. We investigated properties formation of MLV with use of the AHAsomes and Microfluidizer. The multilamellar vesicles are formed merely adding polyol and water phase, followed with the microfluidizer. Formation of a practically pure AHAsomes system, containing the active ingredients directly incorporated without need for preservatives. There were very good encapsulated properties of the active ingredients whether hydrophilic(malic acid, tartaric acid, lactic acid, allantoin, urea) and hydrophobic(vitamin-I acetate, vitamin-A palmitate). Optimum condition (ormatiom of MLV was passed three times in the microfluidizer, particle size of the vesicles should be within range 50-523nm (mean=163.5nm). As application, It was compared that horny layer of the sole of foots removal with the general OM emulsion and the AHAsomes cream. There was used for three months, those got recovery wrinkles about 151.8% and elasticity three times more the AHAsomes than O/W emulsions, It was confirmed with the Image Analyzer and the Cutometer.

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Synthesis of Lactide/Hyaluronic Acid Polymer Membrane for the Application of Drug Delivery System (약물방출시스템 적용을 위한 락타이드/히아루론산 고분자 막의 제조)

  • Kim, Min-Su;Kwon, Ji-Young;Cheong, Seong-Ihl
    • Membrane Journal
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    • v.15 no.4
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    • pp.281-288
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    • 2005
  • The hyaluronic acid (HA) with excellent biocompatibility can be combined with lactide, the ester dimer of polylactide, with good biodegradability to produce biocompatible materials applicable to drug delivery system. By freeze drying method, HA and lactide were crosslinked with crosslinking agent, 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide. Degree of lactide and EDC reaction was determined by the analysis of nuclear magnetic resonance (NMR) spectroscopy. The degree of lactide and EDC reaction increased and swelling ratio decreased as the mole ratio of lactide to HA or crosslinking agent concentration increased or reaction temperature decreased. The drug release experiment result from membranes having different degree of lactide reaction showed that drug release rate reduced in proportion to the degree of lactide reaction. The drug release experiment result from drugs having different hyrodphobicity showed that the more hydrophobic drug was released more slowly.