• 제목/요약/키워드: Functional characterization

검색결과 794건 처리시간 0.027초

Characterization of Clay Minerals in Ranch Pasture

  • Kang, Sangjae;Jang, Jeonghun;Park, Nayun;Park, Junhong;Choi, Seyeong;Park, Man;Lee, Changhee;Lee, Donghoon;Zhang, Yongseon
    • 한국토양비료학회지
    • /
    • 제49권1호
    • /
    • pp.53-59
    • /
    • 2016
  • This study deals with the distribution of the clay minerals separated from clay fractions of ranch pastures in Korea and their chemical and mineralogical properties. Crystalline phases of the clay minerals were identified by powder X-ray diffraction (XRD) pattern and FT-IR spectra, and their relative chemical compositions were also analyzed by X-ray flourescence spectrometry (XRF). Primary minerals consisted mainly of quartz and mica and chlorite and kaolinite along with a trace of swelling micas were identified as secondary clay minerals. However, the relative content of these clay minerals was different with the locations, which led to significant effects on physical and chemical properties of soils like inorganic elemental composition. In particular, $SiO_2$ content was higher in Gochang ranch pasture than in other ranch pasture. Infrared (IR) spectra did not indicate any significant differences in organic functional groups among the locations. This study clearly showed that ranch pastures had different relative content of clay minerals and chemical properties depending on the location and consequently that those properties are worthy to be taken into account for soil amendment.

Fluorescence Microscopy of Condensed DNA Conformations of Bacterial Cells

  • Suleymanoglu, Erhan
    • Journal of Microbiology
    • /
    • 제40권4호
    • /
    • pp.319-326
    • /
    • 2002
  • Cellular DNA in prokaryotes is organized in nucleic acid-protein self-assemblies referred to as the nucleoid. The physical forces responsible for its stability inside the poor solvent properties of the cytoplasm and their functional implications are not understood. Studies on the organisation and functioning of the cytosol of cells largely rely on experimental protocols performed in highly dilute solutions using biochemically purified molecules, which is not a reliable substitute for the situation existing in vivo. Our current research interest is focused on the characterization of biological and physical forces determining the compaction and phase separation of DNA in Escherichia coli cytoplasm. We have emphasized the effect of excluded volume in solutions with high macromolecular concentrations (macromolecular crowding) upon self-association patterns of reactions. The prokaryotic cytosol was simulated by addition of inert polymer polyethylene glycol (PEG) (average molecular weight 20000), as an agent which afterwards facilitates the self-association of macromolecules. Fluorescence microscopy was used for direct visualization of nucleoids in intact cells, after staining with DAPI (4',6-diamidino-2-phenylindole dihydrochloride). Addition of the crowding agent PEG 20,000, in increasing concentrations generated progressively enhanced nucleoid compaction, the effect being stronger in the presence of 0.2 M NaCl and 5 mM MgCl$\_$2/. Under these conditions, the nucleoids were compacted to volumes of around 2 ㎛$\^$3/ or comparable sizes with that of living cells.

Characterization of Single Nucleotide Polymorphisms in 55 Disease-Associated Genes in a Korean Population

  • Lee, Seung-Ku;Kim, Hyoun-Geun;Kang, Jason-J.;Oh, Won-Il;Oh, Berm-Seok;Kwack, Kyu-Bum
    • Genomics & Informatics
    • /
    • 제5권4호
    • /
    • pp.152-160
    • /
    • 2007
  • Most common diseases are caused by multiple genetic and environmental factors. Among the genetic factors, single nucleotide polymorphisms (SNPs) are common DNA sequence variations in individuals and can serve as important genetic markers. Recently, investigations of gene-based and whole genome-based SNPs have been applied to association studies for marker discovery. However, SNPs are so population-specific that the association needs to be verified. Fifty-five genes and 384 SNPs were selected based on association with disease. Genotypes of 337 SNPs in candidate genes were determined using Illumina Sentrix Array Matrix (SAM) chips by an allele-specific extension method in 364 unrelated Korean individuals. Allelic frequencies of SNPs were compared with those of other populations obtained from the International HapMap database. Minor allele frequencies, linkage disequilibrium blocks, tagSNPs, and haplotypes of functional candidate SNPs in 55 genetic disease-associated genes were provided. Our data may provide useful information for the selection of genetic markers for gene-based genetic disease-association studies of the Korean population.

Structural flexibility of Escherichia coli IscU, the iron-sulfur cluster scaffold protein

  • Kim, Bokyung;Kim, Jin Hae
    • 한국자기공명학회논문지
    • /
    • 제24권3호
    • /
    • pp.86-90
    • /
    • 2020
  • Iron-sulfur (Fe-S) clusters are one of the most ancient yet essential cofactors mediating various essential biological processes. In prokaryotes, Fe-S clusters are generated via several distinctive biogenesis mechanisms, among which the ISC (Iron-Sulfur Cluster) mechanism plays a house-keeping role to satisfy cellular needs for Fe-S clusters. The Escherichia coli ISC mechanism is maintained by several essential protein factors, whose structural characterization has been of great interest to reveal mechanistic details of the Fe-S cluster biogenesis mechanisms. In particular, nuclear magnetic resonance (NMR) spectroscopic approaches have contributed much to elucidate dynamic features not only in the structural states of the protein components but also in the interaction between them. The present minireview discusses recent advances in elucidating structural features of IscU, the key player in the E. coli ISC mechanism. IscU accommodates exceptional structural flexibility for its versatile activities, for which NMR spectroscopy was particularly successful. We expect that understanding to the structural diversity of IscU provides critical insight to appreciate functional versatility of the Fe-S cluster biogenesis mechanism.

NMR-based structural characterization of transthyretin in its aggregation-prone state

  • Kim, Bokyung;Kim, Jin Hae
    • 한국자기공명학회논문지
    • /
    • 제24권3호
    • /
    • pp.91-95
    • /
    • 2020
  • Transthyretin (TTR) is an abundant protein in blood plasma and cerebrospinal fluid (CSF), working as a homo-tetrameric complex to transport thyroxine (T4) and a holo-retinol binding protein. TTR is well-known for its amyloidogenic property; several types of systemic amyloidosis diseases are caused by aggregation of either wild-type TTR or its variants, for which more than 100 mutations were reported to increase the amyloidogenicity of TTR. The rate-limiting step of TTR aggregation is the dissociation of a monomeric subunit from a tetrameric complex. A wide range of biochemical and biophysical techniques have been employed to elucidate the TTR aggregation processes, among which nuclear magnetic resonance (NMR) spectroscopy contributed much to characterize the structural and functional features of TTR during its aggregation processes. The present review focuses on discussing the recent advances of our understanding to the amyloidosis mechanism of TTR and to the structural features of its monomeric aggregation-prone state in solution. We expect that the present review provides novel insights to appreciate the molecular basis of TTR amyloidosis and to develop novel therapeutic strategies to treat diverse TTR-related diseases.

From the Sequence to Cell Modeling: Comprehensive Functional Genomics in Escherichia coli

  • Mori, Hirotada
    • BMB Reports
    • /
    • 제37권1호
    • /
    • pp.83-92
    • /
    • 2004
  • As a result of the enormous amount of information that has been collected with E. coli over the past half century (e.g. genome sequence, mutant phenotypes, metabolic and regulatory networks, etc.), we now have detailed knowledge about gene regulation, protein activity, several hundred enzyme reactions, metabolic pathways, macromolecular machines, and regulatory interactions for this model organism. However, understanding how all these processes interact to form a living cell will require further characterization, quantification, data integration, and mathematical modeling, systems biology. No organism can rival E. coli with respect to the amount of available basic information and experimental tractability for the technologies needed for this undertaking. A focused, systematic effort to understand the E. coli cell will accelerate the development of new post-genomic technologies, including both experimental and computational tools. It will also lead to new technologies that will be applicable to other organisms, from microbes to plants, animals, and humans. E. coli is not only the best studied free-living model organism, but is also an extensively used microbe for industrial applications, especially for the production of small molecules of interest. It is an excellent representative of Gram-negative commensal bacteria. E. coli may represent a perfect model organism for systems biology that is aimed at elucidating both its free-living and commensal life-styles, which should open the door to whole-cell modeling and simulation.

The First-principles View of Nanometal Alloy Catalysts

  • Ham, Hyung Chul;Hwang, Gyeong S.
    • 한국진공학회:학술대회논문집
    • /
    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
    • /
    • pp.129-129
    • /
    • 2013
  • Nanometal alloy catalysts have been found to significantly increase catalytic efficiency, compared to the monometallic counterparts. This enhancement can be attributed to various alloying effects: i) the existence of uniquemixed-metal surface sites [the so called ensemble (geometric) effect]; ii) electronic state changes due to metal-metal interactions [the so called ligand (electronic) effect]; and iii) strain caused by lattice mismatch between the alloy components [the socalled strain effect]. In addition, the presence of low-coordination surface atoms and preferential exposure of specific facets [(111), (100), (110)] in association with the size and shape of nanoparticle catalysts [the so called shape-size-facet effect] can be another important factor for modifying the catalytic activity. However, mechanisms underlying the alloying effect still remain unclear owing to the difficulty of direct characterization. Computational approaches, particularly the prediction using first-principles density functional theory (DFT), can be a powerful and flexible alternative for unraveling the role of alloying effects in catalysis since those can give us quantitative insights into the catalytic systems. In this talk, I will present the underlying principles (such as atomic arrangement, facet, local strain, ligand interaction, and effective atomic coordination number at the surface) that govern catalytic reactions occurring on Pd-based alloys using the first-principles calculations. This work highlights the importance of knowing how to properly tailor the surface reactivity of alloy catalysts for achieving high catalytic performance.

  • PDF

Characterization of a Chromosomal Nickel Resistance Determinant from Klebsiella oxytoca CCUG 15788

  • Park, Jae-Sun;Lee, Sung-Jae;Rhie, Ho-Gun;Lee, Ho-Sa
    • Journal of Microbiology and Biotechnology
    • /
    • 제18권6호
    • /
    • pp.1040-1043
    • /
    • 2008
  • Klebsiella oxytoca CCUG 15788 is resistant to $Ni^{2+}$ at a concentration of 10 mM and grows in an inducible manner when exposed to lower concentrations of $Ni^{2+}$. The complete genomic sequence of a 4.2-kb HindIII-digested fragment of this strain was determined from genomic DNA. It was shown to contain four nickel resistance genes (nirA, nirB, nirC, and nirD) encoding transporter and transmembrane proteins for nickel resistance. When the plasmid pKOHI4, encoding nirABCD, was transformed into Escherichia coli JM109, the cells were able to grow in Tris-buffered mineral medium containing 3 mM nickel. TnphoA'-1 insertion mutants in the four nickel genes nirA, nirB, nirC, and nirD showed nickel sensitivity. The nir genes were heterogeneously expressed in E. coli, suggesting functional roles of these genes in nickel resistance.

Molecular Cloning and Expression of Candida antarctica lipase B in Corynebacterium genus

  • Gonzalez, Tamara;M'Barek, Hasna Nait;Gomaa, Ahmed E.;Hajjaj, Hassan;Zhen, Chen;Dehua, Liu
    • 한국미생물·생명공학회지
    • /
    • 제47권4호
    • /
    • pp.546-554
    • /
    • 2019
  • This study, for the first time, reports the functional expression of lipase B derived from the yeast Candida antarctica (CALB) in Corynebacterium strain using the Escherichia coli plasmid PK18. The CALB gene fragment encoding a 317-amino-acid protein was successfully obtained from the total RNA of C. antarctica. CALB was readily produced in the Corynebacterium strain without the use of induction methods described in previous studies. This demonstrated the extracellular production of CALB in the Corynebacterium strain. CALB produced in the Corynebacterium MB001 strain transformed with pEC-CALB recombinant plasmid exhibited maximum extracellular enzymatic activity and high substrate affinity. The optimal pH and temperature for the hydrolysis of 4-nitrophenyl laurate by CALB were 9.0 and 40℃, respectively. The enzyme was stable at pH 10.7 in the glycine-KOH buffer and functioned as an alkaline lipase. The CALB activity was inhibited in the presence of high concentration of Mg2+, which indicated that CALB is not a metalloenzyme. These properties are key for the industrial application of the enzyme.

Expression and Characterization of Trehalose Biosynthetic Modules in the Adjacent Locus of the Salbostatin Gene Cluster

  • Choeng, Yong-Hoon;Yang, Ji-Yeon;Delcroix, Gaetan;Kim, Yoon-Jung;Chang, Yong-Keun;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권10호
    • /
    • pp.1675-1681
    • /
    • 2007
  • The pseudodisaccharide salbostatin, which consists of valienamine linked to 2-amino-1,5-anhydro-2-deoxyglucitol, is a strong trehalase inhibitor. From our Streptomyces albus ATCC 21838 genomic library, we identified thirty-two ORFs in a 37-kb gene cluster. Twenty-one genes are supposed to be a complete set of modules responsible for the salbostatin biosynthesis. Through sequence analysis of the gene cluster, some of the upstream gene products (SalB, SalC, SalD, SalE, and SalF) revealed functional resemblance with trehalose biosynthetic enzymes. On the basis of this rationale, we isolated the five genes (salB, salC, salD, salE, and salF) from the S. albus ATCC 21838 and cloned them into the expression vector pWHM3. We demonstrated the noticeable expression and accumulation of trehalose, using only the five upstream biosynthetic gene cluster of salbostatin, in the transformed Streptomyces lividans TK24. Finally, 490 mg/l trehalose was produced by fermentation of the transformant with sucrosedepleted R2YE media.