• Title/Summary/Keyword: molecular shape

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Cloning and Characterization of Monofunctional Catalase from Photosynthetic Bacterium Rhodospirillum rubrum S1

  • Lee, Dong-Heon;Oh, Duck-Chul;Oh, You-Sung;Malinverni, Juliana C.;Kukor, Jerome J.;Kahng, Hyung-Yeel
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1460-1468
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    • 2007
  • In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

Synthesis and Characterization of Zeolite 4A on Activated Carbon Supports (활성탄 지지체상에서 제올라이트 4A 합성 및 특성)

  • Park, Jeong-Hwan;Suh, Jeong-Kwon;Jeong, Soon-Yong;Lee, Jung-Min;Doh, Myung-Ki
    • Applied Chemistry for Engineering
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    • v.8 no.2
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    • pp.204-210
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    • 1997
  • Zeolite 4A-impregnated complex molecular sieve was prepared by hydrothermal reaction after aluminosilicate gel was penetrated into the pore of activated carbon granule. The crystals of zeolite 4A mainly were formed in the macropore of activated carbon, and their average diameter is $0.8{\mu}m$. The pore volume of activated carbon granule is $0.67m{\ell}/g$, and the pore volume of the sample including 21.6wt% of zeolite 4A crystal is $0.41m{\ell}/g$ decreasing the pore volume by 40% due to the crystallization of zeolite 4A crystals on the internal surface of activated carbon. The calcium ion exchange capacity of zeolite 4A-impregnated sample is 320mg $CaCO_3/g$ zeolite, and this value is almost the same as that of zeolite 4A powder. The crystal of zeolite 4A was not separated from the support of activated carbon granule in the course of ultrasonic dispersion. The adsorption isotherm of water on zeolite 4A-impregnated sample shows the intermediate shape between types, I and III. In addition, zeolite 4A-impregnated sample shows the hydrophilic and hydrophobic properties simultaneously.

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Isolation and Characterization of Colletotrichum Isolates Causing Anthracnose of Japanese Plum Fruit (자두 탄저병균의 분리 및 동정)

  • Lee, Yong-Se;Ha, Da-Hee;Lee, Tae-Yi;Park, Min-Jung;Chung, Jong-Bae;Jeong, Byeong-Ryong
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.299-305
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    • 2017
  • BACKGROUND: Although the filamentous fungal pathogen Colletotrichum species causing anthracnose disease on various fruits including peach, apple, persimmon and grape, there is no report on Japanese plum in Korea. METHODS AND RESULTS: In 2016, diseased fruits showing typical anthracnose symptoms of Japanese plum were collected in market and ochards. Diseased tissue was cut off and disinfected subsequently with 70% ethanol for 1 min, and in 1% sodium hypochloride solution for 1 min, followed by three washes with sterile distilled water. The disinfected tissues were placed onto potato dextrose agar (PDA), and incubated at $25^{\circ}C$ in the dark for 5 to 7 days. For single-spore isolation, conidia were scraped off the plate using a loop, and suspended with 10 mL sterile distilled water. One hundred microliter of the conidial suspension was spread on PDA plates and incubated at $25^{\circ}C$. Finally, one germinated conidium was transferred onto PDA plates. Morphological and cultural characteries of colonies and spores of isolated Colletotrichum were observed after 7 to 10 days incubation on PDA. Molecular identification of isolates were analyzed by comparing rDNA-ITS gene sequences with NCBI GeneBank. CONCLUSION: Of eleven isolates of Colletotrichum isolated from anthracnose diseased Japanese plum fruits, six were identified as C. acutatum, and five as C. gloeosporioides based on diagnostic characteristics such as colony growth rate, shape and size of conidia, and rDNA-ITS sequences. This is the first report of Colletotrichum causing the anthracnose on Japanese plum in Korea.

Embryology of Gymnospermium microrrhynchum (Berberidaceae) (한계령풀의 생식기관 발생형태)

  • Ghimire, Balkrishna;Shin, Dong-Yong;Heo, Kweon
    • Korean Journal of Plant Taxonomy
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    • v.40 no.4
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    • pp.226-233
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    • 2010
  • An intensive study of the embryology of Gymnospermium microrrhynchum was conducted to provide information regarding a discussion of the phylogenetic relationships of the genus, which is yet unstudied. Our results indicated that Gymnospermium is similar to other genera of Berberidaceae in terms of its embryological features. Nevertheless, newly reported and unique features are the well-developed endothelium and the undifferentiated seed coat type. Until the study of Gymnospermium, it may have been considered to be closer to Caulophyllum and Leontice in the tribe Leonticeae. These three genera share many morphological features as well as molecular similarities, by which they are kept in the same tribe, Leonticeae. However, very little detailed embryological data regarding these genera have been published thus far. Gymnospermium was characterized according to the basic type of anther wall formation as well as its glandular tapetum, successive cytokinesis in the microspore mother cell, two-celled mature pollen grains, anatropous and crassinucellate ovules with a nucellar cap, well-developed endothelium, its Polygonum type of embryo sac formation, its nuclear type of endosperm formation, and its undifferentiated seed coat type. In comparison with Nandina, there are many differences, such as the dehiscence of the anther, the cytokinesis in the microspore mother cells, the shape of the megaspore dyad, and the seed characteristics. Although we had no available detailed embryological information regarding Caulophyllum and Leontice, which are genera that are more closely related to Gymnospermium, we could deduce from the phylogenetic relationship that Gymnospermium, Caulophyllum, and Leontice are more closely related to each other than other genera of Berberidaceae on the basis of the seed characteristics.

The Effect of Surfactant on Controlled Release of Amino acids Through Poly(2-Hydroxyethyl Methacrylate) Membrane (Poly(2-Hydroxyethyl Methacrylate)막을 통한 아미노산의 방출 조절에 대한 계면활성제의 효과)

  • Kim Ui-Rak;Jeong Bong-Jin;Lee Myung-Jae;Min Kyung-Sub
    • Journal of the Korean Chemical Society
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    • v.37 no.1
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    • pp.22-35
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    • 1993
  • The transport phenomena of the free amino acids through poly(hydroxyethyl methacrylate)[P(HEMA)] have been investigated with and without various kinds of surfactants solution and in the mixed surfactants solution. Glutamine has the highest diffusivity among 4 amino acids at 1CMC of cetyldimethylethylammonium bromide(CTABr) surfactant. Glutamic acid is not affected by the concentration of CTABr. Methionine and Lysine shows slight decreased diffusivity at 0.5 CMC, but increase its diffusivity at 1CMC and 2CMC due to the structure change of membrane and the viscosity change of surfactant solution. Glutamic acid has the highest diffusivity among four amino acids at sodium dodecyl sulfate(SDS) and Triton X-100 surfactant. In mixed surfactant solution, each amino acids shows high diffusivity through 45% water content membrane at the 0.5 mole fraction of SDS in the SDS/TX-100 surfactant mixtures. It has been found that not only the property of membrane but also the effects of solute-solvent interactions and solvent effect are very important as the permeation of amino acids occurs through P(HEMA) membrane. The diffusivities of free amino acids through membrane depend upon their molecular shape, size and charge.

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Endometrial profilin 1: A key player in embryo-endometrial crosstalk

  • Lee, Chang-Jin;Hong, Seon-Hwa;Yoon, Min-Ji;Lee, Kyung-Ah;Ko, Jung-Jae;Koo, Hwa Seon;Kim, Jee Hyun;Choi, Dong Hee;Kwon, Hwang;Kang, Youn-Jung
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.2
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    • pp.114-121
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    • 2020
  • Objective: Despite extensive research on implantation failure, little is known about the molecular mechanisms underlying the crosstalk between the embryo and the maternal endometrium, which is critical for successful pregnancy. Profilin 1 (PFN1), which is expressed both in the embryo and in the endometrial epithelium, acts as a potent regulator of actin polymerization and the cytoskeletal network. In this study, we identified the specific role of endometrial PFN1 during embryo implantation. Methods: Morphological alterations depending on the status of PFN1 expression were assessed in PFN1-depleted or control cells grown on Matrigel-coated cover glass. Day-5 mouse embryos were cocultured with Ishikawa cells. Comparisons of the rates of F-actin formation and embryo attachment were performed by measuring the stability of the attached embryo onto PFN1-depleted or control cells. Results: Depletion of PFN1 in endometrial epithelial cells induced a significant reduction in cell-cell adhesion displaying less formation of colonies and a more circular cell shape. Mouse embryos co-cultured with PFN1-depleted cells failed to form actin cytoskeletal networks, whereas more F-actin formation in the direction of surrounding PFN1-intact endometrial epithelial cells was detected. Furthermore, significantly lower embryo attachment stability was observed in PFN1-depleted cells than in control cells. This may have been due to reduced endometrial receptivity caused by impaired actin cytoskeletal networks associated with PFN1 deficiency. Conclusion: These observations definitively demonstrate an important role of PFN1 in mediating cell-cell adhesion during the initial stage of embryo implantation and suggest a potential therapeutic target or novel biomarker for patients suffering from implantation failure.

Characteristics of BCNU-loaded PLGA Wafers (BCNU를 함유한 생분해성 PLGA 웨이퍼의 특성분석)

  • 안태군;강희정;이진수;성하수;정제교
    • Polymer(Korea)
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    • v.26 no.5
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    • pp.691-700
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    • 2002
  • Interstitial therapy using biodegradable polymeric device loaded with anticancer agent can deliver the drug to the tumor site at high concentration, resulting in an increase of therapeutic efficacy. 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, carmustine) is most commonly used as chemotherapeutic agent for brain tumors. The design of implantable device is regarded as an important factor lot the efficient delivery of antitumor agent to targeting site. In order to control the release profile of drug, the release pattern of BCNU with the changes of various dimension and additives was investigated. The PLGA wafers containing 3.85, 10, 20 and 30% of BCNU were prepared in various shape (diameter of 3, 5 and 10 mm, thickness of 0.5, 1 and 2 mm) by direct compression method. In vitro drug release profile of BCNU-loaded PLGA wafers could be controlled by changing the dimension of wafers such as initial drug content, weight, diameter, thickness, volume and surface area of wafers, as well as PLGA molecular weight and additives. Drug release from BCNU-loaded PLGA wafers was facilitated with an increase of BCNU-loading amount or presence of poly(N-vinylpyrrolidone)(PVP) or sodium chloride (NaCl). The effects of various geometric factors and additives on the BCNU release pattern were confirmed by the investigation of mass loss and morphology of BCNU-loaded PLGA wafers.

Effect of Thermal History on the Physical Properties of Nylon66 (열 이력이 나일론66의 물성에 미치는 영향)

  • Lee, Bom Yi;Jo, Chan Woo;Shim, Chang Up;Lim, Su Jung;Kim, Youn Cheol
    • Applied Chemistry for Engineering
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    • v.25 no.1
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    • pp.90-95
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    • 2014
  • Nylon66 extrudates as a function of the extrusion number were prepared by a twin screw extruder. Chemical structures, thermal properties, melt index, crystal structures, mechanical properties such as the tensile strength, elongation at break and impact strength, and rheological property were measured by FT-IR, $^1H$-NMR, melt indexer, DSC, TGA, XRD, universal tensile tester, Izod impact tester, and rheometer. FT-IR and $^1H$-NMR characterizations indicated that the number of extrusions did not affect the chemical structure. The decrease in the molecular weight was checked by the melt index of extrudates. There were no effects of the thermal history on the melting and degradation temperature. The tensile and impact strength and modulus were found to be similar, regardless of the number of extrusions, but the elongation decreased significantly. The complex viscosity of extrudates at low frequencies decreased with the extrusion number. No structural changes after extrusion were confirmed from the fact that there was no change in the slope and shape of G'-G" plot.

Isolation of the Protease-producing Yeast Pichia anomala CO-1 and Characterization of Its Extracellular Neutral Protease (세포 외 중성 단백질분해효소를 생산하는 Pichia anomala CO-1의 분리 동정 및 효소 특성)

  • Kim, Ji Yeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1126-1135
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    • 2019
  • From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.

Taxonomic studies of the tribe Potentilleae (Rosaceae) in Korea (한국산 양지꽃족(장미과)의 분류학적 연구)

  • HEO, Kyeong-In;LEE, Sangryong;KIM, Yongsung;PARK, Jongsun;LEE, Sangtae
    • Korean Journal of Plant Taxonomy
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    • v.49 no.1
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    • pp.28-69
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    • 2019
  • We conducted taxonomical investigations based on morphological characteristics, fruit morphology, and literature research on the tribe Potentilleae (Rosaceae) in Korea covering seven genera and 24 species. The style position on the ovary and the shape of style were useful characteristics for the classification of subtribal and generic levels in the tribe Potentilleae. The subtribe Fragariinae is characterized by subbasal or lateral style on the ovary and anthers with one theca. The subtribe Potentillinae has a subterminal style on the ovary, except for Argentina, which presents a subterminal and lateral style and anthers with two thecae. These results support the recent taxonomic recognition that i) the tribe Potentilleae consists of two subtribes, and ii) genera such as Dasiphora, Comarum, and Sibbaldianthe sometimes included in Potentilla s.l. are treated as independent genus. In the subtribe Potentillinae, Argentina, which has been treated as Potentilla, is supported as a distinct genus according to the characteristics of the subterminal and lateral style position and the ventral stipular auricles. In Fragaria, F. nipponica subsp. chejuensis, which has generally small leaves and a limited distribution only on Hallasan Mt., is supported by treatment as an endemic species. Duchesnea chrysantha is distinguished from D. indica by the characteristics of light green or yellowish green leaves, thin and somewhat membranous leaflets, and broad ovate or obovate leaflets. Each complex of P. dickinsii and P. chinensis remains unresolved with regard to controversy over the taxonomic circumscription due to their external morphological variations. Additional taxonomical research and molecular population studies are required for a more in-depth understanding of the tribe Potentilleae in Korea.