• Title/Summary/Keyword: Covalently linked

Search Result 67, Processing Time 0.021 seconds

Subunit Organization of Bacterial Malonate Decarboxylases: The Smallest ${\delta}$ Subunit as an Acyl-Carrier Protein

  • Byun, Hye-Sin;Kim, Yu-Sam
    • BMB Reports
    • /
    • v.30 no.2
    • /
    • pp.132-137
    • /
    • 1997
  • In order to compare molecular structure, malonate decarboxylases from Acinetobacter calcoaceticus, Pseudomonas fluorescens, and Pseudomonas putida aerobically grown on malonate, were purified by the method employing streptomycin sulfate treatment, chromatography with PBE 94 and ${\omega}-aminohexyl$ agarose. Molecular masses were estimated to be 185, 200, and 200 kDa, respectively. All malonate decarboxylases were multimeric enzymes consisting of four different subunits, $2{\alpha},\;1{\beta},\;1{\gamma},\;and\;1{\delta}$. The molecular masses of the Pseudomonas enzyme subunits were $65({\alpha})$, $33({\beta})$, $30({\gamma})$, and $11kDa({\delta})$; which are very similar to those, $65({\alpha})$, $32({\beta})$, $25({\gamma})$, and $11kDa({\delta})$ of Acinetobacter enzyme. The ${\delta}-subunit$ of the active form of the enzymes was acetylated. The acetyl group may form a thioester bond with the thiol group of the prosthetic group covalently linked to the enzyme. It suggests that such molecular organization is common in all malonate decarboxylases.

  • PDF

Screening of Peptides Bound to Anthrax Protective Antigen by Phage Display

  • Kim, Joung-Mok;Park, Hye-Yeon;Choi, Kyoung-Jae;Jung, Hoe-Il;Han, Sung-Hwan;Lee, Jae-Seong;Park, Joon-Shik;Yoon, Moon-Young
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.11
    • /
    • pp.1784-1790
    • /
    • 2006
  • Bacillus anthracis is a causative agent of anthrax. Anthrax toxins are composed of a protective antigen (PA), lethal factor (LF), and edema factor (EF), in which the PA is a central mediator for the delivery of the two enzymatic moieties LF and EF. Therefore, the PA has been an attractive target in the prevention and vaccinization for anthrax toxin. Recently, it has been reported that the molecule consisting of multiple copies of PA-binding peptide, covalently linked to a flexible polymer backbone, blocked intoxification of anthrax toxin in an animal model. In the present study, we have screened novel diverse peptides that bind to PA with a high affinity (picomolar range) from an M13 peptide display library and characterized the binding regions of the peptides. Our works provide a basis to develop novel potent inhibitors or diagnostic probes with a diverse polyvalence.

Characterization of Antibacterial activity and Synthesis of Sulfanilamide Polymer using Crosslinking Agent (가교제를 이용한 Sulfanilamide 중합체의 합성과 항균특성)

  • Kim, Jong-Woan;Yoon, Chul-Hun;Hwang, Sung-Kwy;Kong, Seung-Dae;Lee, Han-Seab
    • Journal of the Korean Applied Science and Technology
    • /
    • v.17 no.1
    • /
    • pp.37-42
    • /
    • 2000
  • Drug delivery system(DDS) have been actively studied for the past twenty years. Dual action agents are unique chemical entities comprised of two different types of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. In spite of the advent of the antibacterial agent the sulfa agents are the most widely used antibacterial agent today. In this study, new antibacterials derivative was synthesized using glutaraldehyde such as crosslinking agent for the purpose of dual-action as DDS study. Antibacterial activity of these new synthetic derivative between their structures and activities were examined by disc diffusion method. As a result, new synthetic derivative exhibited the broad antibacterial activities against Gram(+) and Gram(-) bacilli. Especially, the antibacterial effect of new synthetic derivative against Gram negative(Esherichia. coli) was much stronger than that against Gram positive.

Nucleotide Sequence of 7.2 kb Mitochondrial Linear Plasmid DNA in Pleurotus ostreatus (Pleurotus ostreatus 미토콘드리아의 7.2 kb 선상 플라스미드 염기서열 분석)

  • 윤혜숙;구용범;노정혜
    • Korean Journal of Microbiology
    • /
    • v.37 no.1
    • /
    • pp.37-41
    • /
    • 2001
  • Two linear plasmid-like DNAs, 10.2 kb and 7.2 kb were found in the mitochondria of P. ostreatus. They have covalently linked 5'-terminal proteins in both ends. Two continuous fragments of 4.7 kb and 2.3 kb from 7.2 kb DNA were cloned and sequenced. Two long open reading frames (ORF1; 2982 bp, 993 a.a and ORF2; 2703 bp, 900 a.a) and one short open reading frame(ORF3; 771 bp, 256 a.a) were found in the 7.2 kb plasmid. The putative ORF1 and ORF2 have conserved motifs of DNA polymerases and RNA polymerases, respectively, while the ORF3 has homologous regions with phosphatase from Plasmodium, and also with adhesine from Mycoplasma.

  • PDF

Improvement of Surface Functionalities, Including Allergenicity Attenuation, of Whole Buckwheat Protein Fraction by Maillard-Type Glycation with Dextran

  • Tazawa, Shigeru;Katayama, Shigeru;Hirabayashi, Masahiro;Yamaguchi, Daiki;Nakamura, Soichiro
    • Preventive Nutrition and Food Science
    • /
    • v.19 no.4
    • /
    • pp.327-332
    • /
    • 2014
  • The purpose of the current study was to determine the effects of the introduction of polysaccharide chains onto the molecular surface of buckwheat proteins on buckwheat protein surface functionality. The whole buckwheat protein fraction (WBP) was prepared using 50 mM phosphate buffer (pH 7.5) containing 0.5 M NaCl and covalently linked with 6 kDa, 17.5 kDa, 40 kDa, 70 kDa, or 200 kDa dextran by Maillard-type glycation through controlled dry-heating at $60^{\circ}C$ and 79% relative humidity for two weeks. Conjugation with 40 kDa dextran improved the water solubility and emulsifying properties of WBP without causing a serious loss of available lysine; 84.9% of the free amino groups were conserved. In addition, we found that the introduction of dextran chains onto the molecular surfaces of WBP attenuated the antigenicity of WBP.

Effects of Molecular Weights on the Physico-pharmaceutical Properties of Poly-L-glutamic acid-cytarabine Conjugates

  • Kim, Chong-Kook;Kwon, Kyoung-Ae;Jeong, Eun-Ju;Lee, Myung-Gull
    • Archives of Pharmacal Research
    • /
    • v.12 no.2
    • /
    • pp.88-93
    • /
    • 1989
  • In order to obtain some informations about the effect of molecular weight on the release rate of drug from drug carrier, two types of poly-L-glutamic acid (PLGA)-cytarabine (ara-C) conjugates, PLGA-ara-C:I and PLGA-ara-C:II, were synthesized using two types of PLGA having different average molecular weight, 43,000 and 77,800, respectively. The PLGA-ara-C conjugates were synthesized by mixed anhydride method and found to be covalently linked. Both types of conjugates charged negatively at biological pH. The pH-dependent release rate of ara-C was observed in both cases, and the release rate was accelerated in basic, acidic conditions (the k values were 0.015 $day^{-1}$ at pH 7.0, 0.024 $day^{-1}$ at pH 5.0, and 0.059 $day^{-1}$ at pH 9.0 in the case of PLGA-ara-C:I) and in the presence of pretense. The time required for the release of 16.5% of ara-C from PLGA-ara-C:I were 8 hr and 144 hr in the presence and absence of protease, respectively. Although both types of conjugates showed similar drug substitution ratio, they showed different release rates. Between the two types of conjugates, PLGA-ara-C:II showed the faster release rate (0.030 vs 0.042 $day^{-1}$ in pH 7.4 phosphate buffer solution at $37^{\circ}C$) and the smaller activation energy for the release of drug (12.5 vs 7.7 Kcal/mol) than PLGA-ara-C:I. The characteristic effect of molecular weight on the release rates of PLGA-ara-C conjugates suggests that the drug release rate might be effectively controlled over a prolonged period of time by the combined use of the different types of PLGA-ara-C conjugates having different molecular weights.

  • PDF

Fabrication of Polymer Thin Films on Solid Substrates (고체 기판에 고분자 박막의 고정화)

  • Kim, Min Sung;Jeong, Yeon Tae
    • Applied Chemistry for Engineering
    • /
    • v.21 no.2
    • /
    • pp.200-204
    • /
    • 2010
  • Surface properties are important for determining the functions and uses of materials. So modification of materials with polymer thin films has emerged as an important method to control the physical and chemical properties of the surface layer. We report a simple and effective method to photochemically attach thin polymeric layers to solid surface without chemical derivatization of the substrate and/or the polymer. The system is based on a photoreactive poly(4-vinylpyridine) (P4VP) thin film which is formed on the $SiO_{2}$ surface via spin coating. This substrate is then covered with another polymer film that is reacted with the benzyl radical moieties by UV irradiation. As a result of photochemical reaction, a thin layer of the later polymer is covalently bound to the surface of P4VP. Unbounded polymer is removed by sonication. The thickness of the attached film is a function of the irradiation time and the molecular weight of the polymer. Spatially defined polymer thin films can be fabricated by way of photolithography.

Synthesis and Characterization of Covalently Cross-Linked SPEEK/Cs-substituted MoSiA/Ceria Composite Membranes with MoSiA for Water Electrolysis (MoSiA를 이용한 수전해용 공유가교 SPEEK/Cs-MoSiA/Ceria복합막의 제조 및 성능 연구)

  • SEO, HYUN;SONG, YU-RI;OH, YUN-SUN;MOON, SANG-BONG;CHUNG, JANG-HOON
    • Transactions of the Korean hydrogen and new energy society
    • /
    • v.26 no.6
    • /
    • pp.524-531
    • /
    • 2015
  • To improve the electrochemical and mechanical characteristics, engineering plastic of the sulfonated polyether ether ketone (SPEEK) as polymer matrix was prepared in the sulfonation reaction of polyether ether ketone (PEEK). The SPEEK organic-inorganic blended composite membranes were prepared by sol-gel casting method. It was loaded with the highly dispersed ceria and cesium-substituted molybdosilicic acid (Cs-MoSiA) and 1,4-diiodobutane which was cross-linking agent contents of $10{\mu}L$. Cs-MoSiA was added to increase proton conductivity. Ceria ($CeO_2$) was used as a free radical scavenger which degrade the membrane in polymer electrolyte membrane water elctrolysis (PEMWE). In conclusion, CL-SPEEK/Cs-MoSiA/Ceria 1% composite membrane showed high proton conductivity 0.2104 S/cm at $25^{\circ}C$ which was better than Nafion 117 membrane.

Development of recombinant human chorionic gonadotropin (hCG) using high-density culture technique of suspension-adapted chinese hamster ovary (CHO) cells

  • Na, Kyu-Heum;Kim, Seung-Chul;Seo, Kwang-Seok;Lee, Sung-Hee;Kang, Soo-Hyung
    • 한국생물공학회:학술대회논문집
    • /
    • 2005.04a
    • /
    • pp.37-37
    • /
    • 2005
  • Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

  • PDF

Study of Characteristic of Covalent Cross-linked SPEEK/Silane 4wt%/Cs-substituted MoPA/Ceria hybrid Membrane for Water Electrolysis (Ceria 첨가에 따른 수전해용 공유가교 CL-SPEEK/Silane 4wt%/Cs-MoPA/Ceria 복합막의 특성 연구)

  • Oh, Seunghee;Park, Daeyong;Hwang, Sungha;Yoon, Daejin;Oh, Yunsun;Moon, Sangbong;Chung, Janghoon
    • Transactions of the Korean hydrogen and new energy society
    • /
    • v.25 no.6
    • /
    • pp.561-569
    • /
    • 2014
  • Ceria ($CeO_2$) was used to increase the durability of the membrane in the polymer electrolyte membrane water electrolysis (PEMWE) circumstance. The sulfonated polyether ether ketone (SPEEK) as polymer matrix was prepared in the sulfonation reaction of polyether ether ketone (PEEK) to improve electrochemical characteristics. After sulfonation reaction, the organic-inorganic blended composite membranes were prepared by means of sol-gel casting method with loading the highly dispersed $CeO_2$ and Cs-substituted molybdophosphoric acid (Cs-MoPA) with cross-linking agent (tetrapropyl orthosilicate). Consequently, the composite membrane CL-SPEEK/Silane 4wt%/Cs-MoPA/Ceria(1%) showed the improved characteristics such as 82% of water content, 0.11136 S/cm of proton conductivity at $80^{\circ}C$, 55.50 MPa of tensile strength and 4.37% of breeding out of MoPA.