• Title/Summary/Keyword: secondary structure of ITS

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Characterization of a Biflaviolin Synthase CYP158A3 from Streptomyces avermitilis and Its Role in the Biosynthesis of Secondary Metabolites

  • Lim, Young-Ran;Han, Songhee;Kim, Joo-Hwan;Park, Hyoung-Goo;Lee, Ga-Young;Le, Thien-Kim;Yun, Chul-Ho;Kim, Donghak
    • Biomolecules & Therapeutics
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    • v.25 no.2
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    • pp.171-176
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    • 2017
  • Streptomyces avermitilis produces clinically useful drugs such as avermectins and oligomycins. Its genome contains approximately 33 cytochrome P450 genes and they seem to play important roles in the biosynthesis of many secondary metabolites. The SAV_7130 gene from S. avermitilis encodes CYP158A3. The amino acid sequence of this enzyme has high similarity with that of CYP158A2, a biflaviolin synthase from S. coelicolor A3(2). Recombinant S. avermitilis CYP158A3 was heterologously expressed and purified. It exhibited the typical P450 Soret peak at 447 nm in the reduced CO-bound form. Type I binding spectral changes were observed when CYP158A3 was titrated with myristic acid; however, no oxidative product was formed. An analog of flaviolin, 2-hydroxynaphthoquinone (2-OH NQ) displayed similar type I binding upon titration with purified CYP158A3. It underwent an enzymatic reaction forming dimerized product. A homology model of CYP158A3 was superimposed with the structure of CYP158A2, and the majority of structural elements aligned. These results suggest that CYP158A3 might be an orthologue of biflaviolin synthase, catalyzing C-C coupling reactions during pigment biosynthesis in S. avermitilis.

A New Approach for Practical Classification of Herbicide and for Effective Use by Two-dimensional Ordination Analysis (Two-Dimensional Ordination 분석법에 의한 제초제살초 Spectrum 분류와 효과적인 사용법)

  • Kim Soon Chul
    • Korean journal of applied entomology
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    • v.22 no.2 s.55
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    • pp.147-159
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    • 1983
  • In general, herbicides have been classified according to selectivity, mobility. time of application, methods of application, mode of action and chemical property and structure. However, there was no generally accepted classification system for practical use in the field. The primary processes affected by the majority of herbicides are the growth process through cell elongation and/or cell division, the photosynthetic process specifically the light reaction, the oxidative phosphorylation and the integrity of the membrane systems. The usual approach in the study of the mechanism by which herbicides kill or inhibit the growth of plants is to initially determine the morphological phototoxicity systems, The mechanism by which a herbicide kills a plant or suppresses its development is actually the resultant effect of primary and secondary(or side) effects. In most instances, the death of the plant is due to the secondary effects. To induce the desired response, a herbicide must be able to gain entry into the plants and once inside, to be transported within the plant to its site(s) of activity in concentrations great enough. Obstacles to the entry and movement of herbicides in plants are generally classified by leaf and soil obstacles, translocation obstacles and biochemical obstacles, and these obstacles are also strongly influenced by plant species and by environmental factors such as light, temperature, rainfall and relative humidity. And hence, in most instances, results obtained from laboratory or greenhous vary from those of field experiment. Author attempted to classify herbicides from the field experiment using the two-dimensional ordination analysis to obtain practical information for selecting effective herbicides or to choose effective herbicide combinations for increasing herbicidal efficacy or reducing the chemical cost. Based on this two-dimensional diagram, desired herbicides or combinations were selected and further investigated for the interaction effects whether these combinations are synergistic, additive or antagonistic. From the results, it was concluded that these new approach could possibly be give more comprehensive informations about effective use of herbicide than any other systems.

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Study of the Electrochemical Properties of Li4Ti5O12 Doped with Ba and Sr Anodes for Lithium-Ion Secondary Batteries

  • Choi, Byung-Hyun;Lee, Dae-Jin;Ji, Mi-Jung;Kwon, Young-Jin;Park, Sung-Tae
    • Journal of the Korean Ceramic Society
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    • v.47 no.6
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    • pp.638-642
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    • 2010
  • The spinel material $Li_4Ti_5O_{12}$ has attracted considerable attention as an anode electrode material for many battery applications owing to its light weight and high energy density. However, the real capacity of $Li_4Ti_5O_{12}$ powder as determined by the solid-state method is lower than the ideal capacity. In this study, we investigated the effect of the dopants in M-doped spinel $Ba_xLi_{4-2x}Ti_5O_{12}$(x=0.005, 0.05, 0.1) powders prepared by the solid-state reaction method and used as the anode material in lithiumion batteries. The results confirmed the effect of the Ba and Sr dopants on the powder properties of the spinel $Li_4Ti_5O_{12}$, which exhibited a pure spinel structure without any secondary phase in its XRD pattern. Moreover, the electrochemical properties of the spinel M-LTO materials were investigated using a half cell. The electrochemical data show that cells with anodes made of undoped $Li_4Ti_5O_{12}$ and Ba- and Sr-doped $Li_4Ti_5O_{12}$ have discharge capacities of 97, 130, and 112 mAh/g, respectively, at the first cycle. Moreover, the Ba- and Sr-doped spinel $Li_4Ti_5O_{12}$ demonstrated good properties in the mid-voltage range at 1.55 V, showing stable cyclic voltammogram properties which surpassed those of the same material without Ba or Sr at 1 C after 100 cycles.

Cadmium-Substituted Concanavalin A and Its Trimeric Complexation

  • Park, Yeo Reum;Kim, Da Som;Lee, Dong-Heon;Kang, Hyun Goo;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2106-2112
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    • 2018
  • Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). In this study, ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that $Mn^{2+}$ was replaced by $Cd^{2+}$ (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with $Cd^{2+}$ in this study. The trimeric ConA in this study coordinated $Cd^{2+}$ with other residues, including Asp80 and Asp82, for complex generation. The monomer does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary $Cd^{2+}$ and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.

Microbial Community Structure of Korean Cabbage Kimchi and Ingredients with Denaturing Gradient Gel Electrophoresis

  • Hong, Sung Wook;Choi, Yun-Jeong;Lee, Hae-Won;Yang, Ji-Hee;Lee, Mi-Ai
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1057-1062
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    • 2016
  • Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341FGC-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.

A Study on the Furniture Design of the Early Modernism in the Original Formative-World (초기 모더니즘 가구디자인의 근원적 조형세계)

  • Choi, Byung-Hoon;Lee, Young-Choon
    • Journal of the Korea Furniture Society
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    • v.19 no.5
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    • pp.328-340
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    • 2008
  • This study examined the characteristics of furniture design in the early Modernism that occurred in the early 20th Century through the relationship with modern arts. The furniture design of the early Modernism was established in the early 20th Century based on the simplicity and honesty of the Art and Crafts Movement and Anti-Historicism in Art Nouveau. During this period, the necessity of mass production became critical due to the radical industrialization and new social demands. In such periodic stream, the furniture design of the early modernism pursued simplicity and geometric beauty based on functionalism. The efforts to discover the fundamental structure of furniture were intensified. It was not limited to furniture design but was also exercised in the huge periodic transformation that progressed in every field of art including architecture. The Modernism art has also been connected to Cubism and spread into an abstract direction. While questioning the potential essence and progressing the study on genuine characteristics, the Modernism art tried to return things into their substantial looks and reorganize them conceptually, rather than reproducing external objects. The furniture that was secondary to a part of architecture and interior accessory transcended its decorative purpose. It pursued the structure to follow its fundamental purpose as furniture. Such tendency corresponded to the direction that Post-Modernism followed. Likewise, both art and design had the revivable and abstract characteristics based on the identical objective in ideology. For this reason, the formative and geometric characteristics of the early modernism furniture design have a mutual relationship with modern arts. Particularly, such tendency intensified and progressed through the Bauhaus in Germany. Based on such facts, this study proved that the early Modernism furniture design in the 20th Century tended to appear in common arts including the art and design according to the social demand of the huge periodic stream. Furthermore, the ideology that was adopted in art and design, as well as its formative characteristics, was examined through the mutual relationship of modern arts and design.

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Catalytic Properties of Monomeric Species of Brain Pyridoxine-5'-phosphate Oxidase

  • Kwon, Oh-Shin;Choi, Soo-Young
    • BMB Reports
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    • v.34 no.1
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    • pp.21-27
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    • 2001
  • The structural stability of brain pyrydoxine-5'-phosphate (PNP) oxidase and the catalytic properties of the monomeric species were investigated. The unfolding of brain pyridoxine-5'-phosphate (PNP) oxidase by guanidine hydrochloride (GuHCl) was monitored by means of fluorescence and circular dichroism spectroscopy Reversible dissociation of the dimeric enzyme into subunits was attained by the addition of 2 M GuHCl. The perturbation of the secondary structure under the denaturation condition resulted in the release of the cofactor FMN. Separation of the processes of refolding and reassociation of the monomeric species was achieved by the immobilization method. Dimeric PNP oxidase was immobilized by the covalent attachment to Affi-gel 15 without any significant lass of its catalytic activity. Matrix-bound monomeric species were obtained from the reversible refolding processes. The matrix bound-monomer was found to be catalytically active, possessing only a slightly decreased specific activity when compared to the refolded dimeric enzyme. In addition, limited chymotrypsin digestion of the oxidase yields two fragments of 12 and 161 kDa with a concomitant increase of catalytic activity The catalytically active fragment was isolated by ion exchange chromatography and analyzed for association of two subunits using the FPLC gel filtration analysis. The retention time indicated that the catalytic fragment of 16 kDa behaves as a compact monomer. Taken together, these results are consistent with the hypothesis that the native quaternary structure of PNP oxidase is not a prerequisite for catalytic function, but it could play a role in the regulation.

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Protective Effect of Biological Osmolytes against Heat- and Chaotropic Agent-Induced Denaturation of Bacillus licheniformis γ-Glutamyl Transpeptidase

  • Lo, Huei-Fen;Chi, Meng-Chun;Lin, Min-Guan;Lan, Yuan-Gin;Wang, Tzu-Fan;Lin, Long-Liu
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1457-1466
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    • 2018
  • In the present study, the stabilizing effect of four different biological osmolytes on Bacillus licheniformis ${\gamma}$-glutamyl transpeptidase (BlGGT) was investigated. BlGGT appeared to be stable under temperatures below $40^{\circ}C$, but the enzyme retained less than 10% of its activity at $60^{\circ}C$. The tested osmolytes exhibited different degrees of effectiveness against temperature inactivation of BlGGT, and sucrose was found to be the most effective among these. The use of circular dichroism spectroscopy for studying the secondary structure of BlGGT revealed that the temperature-induced conformational change of the protein molecule could be prevented by the osmolytes. Consistently, the molecular structure of the enzyme was essentially conserved by the osmolytes at elevated temperatures as monitored by fluorescence spectroscopy. Sucrose was further observed to counteract guanidine hydrochloride (GdnHCl)-and urea-induced denaturation of BlGGT. Taken together, we observed evidently that some well-known biological osmolytes, especially sucrose, make a dominant contribution to the structural stabilization of BlGTT.

Backbone 1H, 15N, and 13C Resonance Assignments and Secondary-Structure of the Conserved Hypothetical Protein HP0892 of Helicobacter pylori

  • Han, Kyung-Doo;Park, Sung-Jean;Jang, Sun-Bok;Lee, Bong-Jin
    • Molecules and Cells
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    • v.25 no.1
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    • pp.138-141
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    • 2008
  • HP0892 (SwissProt/TrEMBL ID O25552) is a 90-residue conserved hypothetical protein from Helicobacter pylori strain 26695, with a calculated pI of 9.38 and a molecular mass of 10.41 kDa. It belongs to the Plasmid stabilization system protein family (PF05016) in the Pfam database. Proteins with sequence similarity to HP0892 exist in Vibrio choierae, Enterococcus faecalis, Campylobacter jejuni, Streptococcus pneumoniae, Haemophilus influenzae, Escherichia coli O157. Here we report the sequence-specific backbone resonance assignments of HP0892 using multidimentional heteronuclear NMR spectroscopy. About 97.0% (422/435) of the HN, N, CO, $C{\beta}$, $C{\alpha}$ resonances of 90 residues of HP0892 were assigned. On the basis of the resonance assignments, three helical regions and four strand regions were identified using the CSI program. This study is a prerequisite for calculating the solution structure of HP0892, and will be useful for studying its interaction with other molecules.

Spermatogenesis and its fine structure of the seminiferous epithelium in the Jindo dog (진도견(珍島犬) 정세관상피(精細管上皮)의 정자발생(精子發生)과 미세구조(微細構造))

  • Kim, Yong-hwan;Park, Young-seok
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.23-36
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    • 1993
  • To investigate the cycle and relative frequences and the fine structure of seminiferous epithelia in mature Jindo dogs, histologic study was performed. The results obtained were summarized as follows; 1. Type A spermatogonia appeared approximately 1.6 times as many at stage II as compared to stage I while type In spermatogonia appeared small amount in stage III, IV and V. type B spermatogonia were found during the stage VI to VIII, though not detectable during stage I to V. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage VIII. The number of primary spermatocytes of the leptotene phase markedly increased during stage I to II, and the primary spermatocytes of the pachytene phase were shown the least in number at stage IV. The secondary spermatocytes could be seen only at stage IV. 2. The relative frequencies of each stage from stages I to VIII of the cycle of seminiferous epithelia were 31.6, 11.9, 10.0, 3.2, 8.2, 10.1, 11.7 and 13.2% respectively. 3. On electron microscopic observations, acrosomal vesicle of spermatids appeared larger though the bulk of germ cells were the morphologically same as those of the other animal species. Thread line structures light microscopically observed in the cytoplasm of Sertoli cell were the longitudinal orientation of mitochondria.

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