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A New Esterase, Belonging to Hormone-Sensitive Lipase Family, Cloned from Rheinheimera sp. Isolated from Industrial Effluent

  • Virk, Antar Puneet;Sharma, Prince;Capalash, Neena
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.667-674
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    • 2011
  • The gene for esterase (rEst1) was isolated from a new species of genus Rheinheimera by functional screening of E. coli cells transformed with the pSMART/HaeIII genomic library. E. coli cells harboring the esterase gene insert could grow and produce clear halo zones on tributyrin agar. The rEst1 ORF consisted of 1,029 bp, corresponding to 342 amino acid residues with a molecular mass of 37 kDa. The signal P program 3.0 revealed the presence of a signal peptide of 25 amino acids. Esterase activity, however, was associated with a homotrimeric form of molecular mass 95 kDa and not with the monomeric form. The deduced amino acid sequence showed only 54% sequence identity with the closest lipase from Cellvibrio japonicus strain Ueda 107. Conserved domain search and multiple sequence alignment revealed the presence of an esterase/ lipase conserved domain consisting of a GXSXG motif, HGGG motif (oxyanion hole) and HGF motif, typical of the class IV hormone sensitive lipase family. On the basis of the sequence comparison with known esterases/ lipases, REst1 represents a new esterase belonging to the class IV family. The purified enzyme worked optimally at $50^{\circ}C$ and pH 8, utilized pNP esters of short chain lengths, and showed best catalytic activity with p-nitrophenyl butyrate ($C_4$), indicating that it was an esterase. The enzyme was completely inhibited by PMSF and DEPC and showed moderate organotolerance.

Isolation and Characterization of Some Promoter Sequences from Leuconostoc mesenteroides SY2 Isolated from Kimchi

  • Park, Ji Yeong;Jeong, Seon-Ju;Kim, Jeong A;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1586-1592
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    • 2017
  • Some promoters were isolated and characterized from the genome of Leuconostoc mesenteroides SY2, an isolate from kimchi, a Korean traditional fermented vegetable. Chromosomal DNA of L. mesenteroides SY2 was digested with Sau3AI and ligated with BamHI-cut pBV5030, a promoter screening vector containing a promoterless cat-86. Among E. coli transformants (TFs) resistant against Cm (chloramphenicol), 17 were able to grow in the presence of $1,000{\mu}g/ml$ Cm and their inserts were sequenced. Transcription start sites were examined for three putative promoters (P04C, P25C, and P33C) by primer extension. Four putative promoters were inserted upstream of a promoterless ${\alpha}$-amylase reporter gene in $pJY15{\alpha}$. ${\alpha}$-Amylase activities of E. coli TFs containing $pJY15{\alpha}$ (control, no promoter), $pJY03{\alpha}$ ($pJY15{\alpha}$ with P03C), $pJY04{\alpha}$ (with P04C), $pJY25{\alpha}$ (with P25C), and $pJY33{\alpha}$ (with P33C) were 66.9, 78.7, 122.1, 70.8, and 99.3 U, respectively. Cells harboring $pJY04{\alpha}$ showed 1.8 times higher activity than the control. Some promoters characterized in this study might be useful for construction of food-grade expression vectors for Leuconostoc sp. and related lactic acid bacteria.

Multiple Signaling Molecules are Involved in Expression of CCL2 and IL-$1{\beta}$ in Response to FSL-1, a Toll-Like Receptor 6 Agonist, in Macrophages

  • Won, Keunsoo;Kim, Sun-Mi;Lee, Sae-A;Rhim, Byung-Yong;Eo, Seong-Kug;Kim, Koanhoi
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.6
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    • pp.447-453
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    • 2012
  • TLR6 forms a heterodimer with TLR2 and TLR4. While proinflammatory roles of TLR2 and TLR4 are well documented, the role of TLR6 in inflammation is poorly understood. In order to understand mechanisms of action of TLR6 in inflammatory responses, we investigated the effects of FSL-1, the TLR6 ligand, on expression of chemokine CCL2 and cytokine IL-$1{\beta}$ and determined cellular factors involved in FSL-1-mediated expression of CCL2 and IL-$1{\beta}$ in mononuclear cells. Exposure of human monocytic leukemia THP-1 cells to FSL-1 resulted not only in enhanced secretion of CCL2 and IL-$1{\beta}$, but also profound induction of their gene transcripts. Expression of CCL2 was abrogated by treatment with OxPAPC, a TLR-2/4 inhibitor, while treatment with OxPAPC resulted in partially inhibited expression of IL-$1{\beta}$. Treatment with FSL-1 resulted in enhanced phosphorylation of Akt and mitogen-activated protein kinases and activation of protein kinase C. Treatment with pharmacological inhibitors, including SB202190, SP6001250, U0126, Akt inhibitor IV, LY294002, GF109203X, and RO318220 resulted in significantly attenuated FSL-1-mediated upregulation of CCL2 and IL-$1{\beta}$. Our results indicate that activation of TLR6 will trigger inflammatory responses by upregulating expression of CCL2 and IL-$1{\beta}$ via TLR-2/4, protein kinase C, PI3K-Akt, and mitogen-activated protein kinases.

Variation of Phytoplankton and Nutrients in the Namdaechon Estuary, Korea (남대천 하구역의 식물플랑크톤과 영양염 변동)

  • KWON Kee-Young;KIM Ju-Kyoung;HONG Gwan-Eui;SEONG Ki-Baek;LEE Chul-Ho;MOON Chang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.1
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    • pp.55-64
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    • 2005
  • Spatio-temporal variations of phytoplankton abundance and nutrient concentrations were investigated in the Namdaechon estuary, Yangyang, from April to December, 2003. A total of 51 phytoplankton species were identified with 32 diatom species in the study area. Phytoplankton abundance ranged from 14 cells/mL to 3,798 cells/mL. Small sized benthic and pennate-type diatoms like Cymbella spp., Fragilaria spp., Navicula spp., Synedra spp. were dominant at throughout the whole study area. Various planktonic species like Chlamydomonas sp., Peridinium spp., Euglena spp., Cryptomonas spp. etc. were abundant especially at the estuary of the Namdaechon, from May to August. Phytoplankton bloom (>3,000 \;cells/mL) occurred from the late May to mid June. After September when the sandbank of estuary was broken by the typhoon 'Maemi', planktonic species disappeared. These planktonic species were followed by oceanic diatoms, Skeletonema costatum and Chaetoceros spp.. Concentrations of dissolved inorganic nitrogen (DIN) and silicate were relatively high at the upstream in summer. In May, before phytoplankton bloom, high concentrations of phosphate $(>3.5\;{\mu}mol/L)$ were observed at all the study area. These results suggested that spatio-temporal variation of phytoplankton in the Namdaechon estuary was related to formation of sandbank at the mouth of the estuary, fluid speed affected by sandbank and nutrients supplied in spring. High correlation $(r^2=0.928)$ between chlorophyll a and biological oxygen demand (BOD) implies that BOD was related to phytoplankton abundance in the Namdaechon estuary.

An Approach to Constructing an Efficient Entropy Source on Multicore Processor (멀티코어 환경에서 효율적인 엔트로피 원의 설계 기법)

  • Kim, SeongGyeom;Lee, SeungJoon;Kang, HyungChul;Hong, Deukjo;Sung, Jaechul;Hong, Seokhie
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.28 no.1
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    • pp.61-71
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    • 2018
  • In the Internet of Things, in which plenty of devices have connection to each other, cryptographically secure Random Number Generators (RNGs) are essential. Particularly, entropy source, which is the only one non-deterministic part in generating random numbers, has to equip with an unpredictable noise source(or more) for the required security strength. This might cause an requirement of additional hardware extracting noise source. Although additional hardware resources has better performance, it is needed to make the best use of existing resources in order to avoid extra costs, such as area, power consumption. In this paper, we suggest an entropy source which uses a multi-threaded program without any additional hardware. As a result, it reduces the difficulty when implementing on lightweight, low-power devices. Additionally, according to NIST's entropy estimation test suite, the suggested entropy source is tested to be secure enough for source of entropy input.

Taxonomic Revision of the Lichen Genera Pertusaria, Varicellaria, and Variolaria (Pertusariales, Ascomycota) in South Korea

  • Park, Jung Shin;Park, Sook-Young;Park, Chan-Ho;Kondratyuk, Sergii Y.;Oh, Soon-Ok;Hur, Jae-Seoun
    • Mycobiology
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    • v.45 no.4
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    • pp.270-285
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    • 2017
  • The crustose lichen genus Pertusaria comprises over ca. 800 species worldwide. In total, 20 Pertusaria species were localized to the Mt. Sorak and Jeju-do in Korea. To date, information regarding the distribution of Pertusaria species in the South Korean peninsula is scarce. In this study, we collected Pertusaria species across South Korea and identified them based on morphological, chemical, and molecular characteristics. Of the 387 samples collected, we identified 24 taxa and 1 variety, of which 17 were previously recorded, and 6 taxa were newly found in South Korea (P. leioplaca, P. leucosora var. violascens, P. texana, P. thiospoda, P. thwaitesii, and P. xanthodes), 2 known species were transferred to Varicellaria (Varicellaria lactea and V. velata), one species was transferred to Variolaria as a new record (Variolaria multipunctoides) and one was a new species (P. jogyeensis J. S. Park & J.-S. Hur, sp. nov.). Characteristics of the newly discovered species, P. jogyeensis, are as follows: smooth to bumpy thallus, scattered to crowded poriform apothecia, blackish ostioles, definitely sunken, thin yellowish green rims around ostioles, 8-spored ascus, and the presence of perlatolic acid and thiophaninic acid (chlorinated xanthone). Phylogenetic studies on P. jogyeensis based on the mitochondrial small subunit sequence revealed proximity to P. flavicans and P. texana, and supported its classification as a new species within the genus Pertusaria. Additionally, we describe the chemical composition and morphology of all listed species in detail and provide an artificial key for identification.

Arthothelium punctatum (Arthoniaceae, Arthoniales), A New Lichen Species from South Korea

  • Park, Jung Shin;Park, Sook-Young;Park, Chan-Ho;Jang, Seol-Hwa;Hur, Jae-Seoun
    • Mycobiology
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    • v.45 no.4
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    • pp.255-262
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    • 2017
  • A total of 121 species of lichens belonging to the genus Arthothelium have been described to date, most of which have been found in tropical regions. Here, we describe the discovery of a novel Arthothelium species for the first time in South Korea. Until now, Arthothelium ruanum was the only Arthothelium species reported in South Korea. Among the 113 specimens collected in this study, we identified A. ruanum and a putative new species, Arthothelium punctatum (J. S. Park & J.-S. Hur, sp. nov.). The diagnostic characters of A. punctatum are as follows: apothecia punctate, shortly elongate to branched, small, 0.1-0.2 mm wide, hypothecium hyaline to pale brown and obovate to broadly ellipsoid, muriform ascospores, $29.5-44.6{\times}12.2-18.2{\mu}m$. The new species was found in Mt. Seokbyeong at an altitude of 790 m on smooth bark. Upon phylogenic analysis, the putative new species, A. punctatum, was separated from other Arthothelium species although the specimens analyzed were clustered with Arthoniaceae in phylogenetic trees based on both the mitochondrial small subunit (mtSSU) sequence and combined mtSSU and nuclear ribosomal large subunit sequences. Our data clearly indicate that this species is a new species belonging to the family Arthoniaceae. To elucidate the taxonomic characteristics of the new species, we provide morphological descriptions and a distribution map.

Assessment of Pathogenic Variation against Pitch Canker Pathogen, Fusarium circinatum in Pinus thunbergii and Responses of Natural Selection Pinus × rigitaeda to Branch Inoculation in a Seed Orchard

  • Woo, Kwan-Soo;Yoon, Jun-Hyuck;Han, Sang-Urk;Kim, Chang-Soo
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.299-305
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    • 2010
  • A half-sib family of two 4-year-old seedlings of Pinus $\times$ rigitaeda was inoculated with each of 20 Pinus thunbergii isolates of Fusarium circinatum (syn. Fusarium subglutinans f. sp. pini) from two pitch canker damaged sites in Jeju Island, South Korea. Initial symptoms of needle damages were visible on most of the seedlings at 18 days after inoculation. The 20 tested isolates were not significantly different in virulence, based on lesion lengths at the site of inoculation (P = 0.217). The most virulent isolate FT-7 showed the longest lesion length. Some seedlings began to die 46 days after inoculation. All seedlings were dead by 68 days after inoculation except two seedlings inoculated with each of isolates FS-2 and FS-13, respectively. Using the FT-7, 38-year-old 11 P. $\times$ rigitaeda trees, which were survived from a seed orchard severely damaged by pitch canker, were inoculated on branches in the seed orchard in Jeju Island to assess differences in susceptibility to pitch canker. The 11 trees differed significantly (P < 0.001) in susceptibility to F. circinatum based on average lesion lengths measured 56 days after inoculation. It is possible that induced resistance contributed to their capacity to limit lesion development. The susceptibility of natural selection P. $\times$ rigitaeda trees are more likely affected by interaction with F. circinatum rather than environmental conditions.

Cyclic Dipeptides from Bacillus vallismortis BS07 Require Key Components of Plant Immunity to Induce Disease Resistance in Arabidopsis against Pseudomonas Infection

  • Noh, Seong Woo;Seo, Rira;Park, Jung-Kwon;Manir, Md. Maniruzzaman;Park, Kyungseok;Sang, Mee Kyung;Moon, Surk-Sik;Jung, Ho Won
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.402-409
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    • 2017
  • Cyclic dipeptides (CDPs) are one of the simplest compounds produced by living organisms. Plant-growth promoting rhizobacteria (PGPRs) also produce CDPs that can induce disease resistance. Bacillus vallismortis strain BS07 producing various CDPs has been evaluated as a potential biocontrol agent against multiple plant pathogens in chili pepper. However, plant signal pathway triggered by CDPs has not been fully elucidated yet. Here we introduce four CDPs, cyclo(Gly-L-Pro) previously identified from Aspergillus sp., and cyclo(L-Ala-L-Ile), cyclo(L-Ala-L-Leu), and cyclo(L-Leu-L-Pro) identified from B. vallismortis BS07, which induce disease resistance in Arabidopsis against Pseudomonas syringae infection. The CDPs do not directly inhibit fungal and oomycete growth in vitro. These CDPs require PHYTOALEXIN DEFICIENT4, SALICYLIC ACID INDUCTION DEFICIENT2, and NONEXPRESSOR OF PATHOGENESIS-RELATED PROTEINS1 important for salicylic acid-dependent defense to induce resistance. On the other hand, regulators involved in jasmonate-dependent event, such as ETHYLENE RECEPTOR1, JASMONATE RESPONSE1, and JASMONATE INSENSITIVE1, are necessary to the CDP-induced resistance. Furthermore, treatment of these CDPs primes Arabidopsis plants to rapidly express PATHOGENESIS-RELATED PROTEIN4 at early infection phase. Taken together, we propose that these CDPs from PGPR strains accelerate activation of jasmonate-related signaling pathway during infection.

Thrips in Medicinal Crops in Korea:Identification and Their Damages (국내 주요 약용작물에 발생하는 총채벌레의 종류와 피해)

  • Kang, Taek Jun;Ahn, Seung Joon;An, Tae Jin;Cho, Myoung Rae;Jeon, Heung Yong;Jung, Jae A
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.487-492
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    • 2012
  • Damages of medicinal crops by thrips and identification of thrips species were investigated by periodic surveys on major medicinal crops, Platycodon grandiflorum, Schizandra chinensis, Codonopsis lanceolata, Lycium chinense, Bupleurum falcatum, Ledebouriella seseloides, Angelica gigas, Glycyrrhiza uralensis, Liriope platyphylla, Atractyloides sp., once or twice a month in Suwon, Eumseong, Jecheon, Pyeongchang, Chyungyang from May to October in 2012. Thrips and their damages were found at all of the medicinal crops investigated, mostly occurring at early growing stages of each medicinal crop and causing apical meristem region withered, thereby inhibiting the growth of plants. For species identification of thrips collected from each medicinal crops, DNA from each individual was extracted and ITS2 and COI regions were amplified by PCR. As a result, Frankliniella intonsa, Thrips tabaci, and Megalurothrips distalis have been identified as dominant species on medicinal crops in open field, whereas Frankliniella occidentalis was restricted to Lycium chinense in which protected cultivation. This study suggests that the occurrence of thrips species on medicinal crops seems to be affected by host specificity and environmental factors such as cultivation types.