• Title/Summary/Keyword: biological activity

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Temporal and Spatial Downregulation of Arabidopsis MET1 Activity Results in Global DNA Hypomethylation and Developmental Defects

  • Kim, Minhee;Ohr, Hyonhwa;Lee, Jee Woong;Hyun, Youbong;Fischer, Robert L.;Choi, Yeonhee
    • Molecules and Cells
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    • v.26 no.6
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    • pp.611-615
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    • 2008
  • DNA methylation is an epigenetic mechanism for gene silencing. In Arabidopsis, MET1 is the primary DNA methyltransferase that maintains CG DNA methylation. Plants having an overall reduction of MET1 activity, caused by a met1 mutation or a constitutively expressed MET1 antisense gene, display genome hypomethylation, inappropriate gene and transposon transcription, and developmental abnormalities. However, the effect of a transient reduction in MET1 activity caused by inhibiting MET1 expression in a restricted set of cells is not known. For this reason, we generated transgenic plants with a MET1 antisense gene fused to the DEMETER (DME) promoter (DME:MET1 a/s). Here we show that DME is expressed in leaf primordia, lateral root primoridia, in the region distal to the primary root apical meristem, which are regions that include proliferating cells. Endogenous MET1 expression was normal in organs where the DME:MET1 a/s was not expressed. Although DME promoter is active only in a small set of cells, these plants displayed global developmental abnormalities. Moreover, centromeric repeats were hypomethylated. The developmental defects were accumulated by the generations. Thus, not maintaining CG methylation in a small population of proliferating cells flanking the meristems causes global developmental and epigenetic abnormalities that cannot be rescued by restoring MET1 activity. These results suggest that during plant development there is little or no short-term molecular memory for reestablishing certain patterns of CG methylation that are maintained by MET1. Thus, continuous MET1 activity in dividing cells is essential for proper patterns of CG DNA methylation and development.

Investigation of Siderophore production and Antifungal activity against Phytophthora capsici as related to Iron (III) nutrition by Lysobacter antibioticus HS124

  • Ko, Hyun-Sun;Tindwa, Hamisi;Jin, Rong De;Lee, Yong-Seong;Hong, Seong-Hyun;Hyun, Hae-Nam;Nam, Yi;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.4
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    • pp.650-656
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    • 2011
  • Lysobacter antibioticus HS124 isolated from pepper rhizosphere soil produced catechol type siderophore. Purified siderophore by Diaion HP-20 and silica gel column chromatography showed several hydroxyl functional groups adjacent to benzene rings by analysis of $^1H$ NMR spectroscopy. The strain HS124 showed different activities to suppress Phytophthora capsici with different concentrations of exogenous Fe (III) in minimal medium where antifungal activity with $100{\mu}M$ Fe (III) was approximately 1.5 times higher than in absence of Fe (III). Bacterial population in this Fe (III)-amended medium was also highest with $8.9{\times}10^8\;CFU\;ml^{-1}$ which also corresponded to the strongest siderophore activity. When grown in rich medium (minimal medium with N, $P_2O_5K_2O$ and glucose), HS124 exhibited approximately 2 times stronger antifungal activity compared to minimal medium. In pot trials, treatments of bacterial culture grown in rich medium with (C1) or without (C2) $100{\mu}M$ Fe (III) exhibited a high protection of pepper plants from disease, compared to medium only with (M1) or without (M2) $100{\mu}M$ Fe (III). Especially, treatment C1 showed the best disease control effect of about 70 %. Thus, the strain HS124 should be recommended as a potential biocontrol agent against P. capsici in pepper.

Biological Activities of Phellinus linteus Mycelium Culture with Cassiae Semen Extract on β-Glucuronidase Inhibitory Activity (β-Glucuronidase 저해 활성이 우수한 결명자를 첨가한 상황 균사체 배양액의 생리활성)

  • Oh, Eun-Hee;Park, Jung-Mi;Kim, Sang-Hee;Song, In-Gyu;Han, Nam-Soo;Yoon, Hyang-Sik
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.620-628
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    • 2012
  • We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were $30^{\circ}C$, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and ${\beta}$-glucan contents also increased, but final pH was not influenced. In addition, the ${\beta}$-glucuronidase inhibitory activity, electron donating activity, and ${\alpha}$-glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

Comparative Analysis of Src Activity in Plasma Membrane Subdomains via Genetically Encoded FRET Biosensors (유전적으로 암호화된 FRET 바이오센서를 통한 세포막 하위 도메인의 Src 활성 비교 분석)

  • Gyuho Choi;Yoon-Kwan Jang;Jung-Soo Suh;Heonsu Kim;Sanghyun Ahn;Tae-Jin Kim
    • Journal of Life Science
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    • v.33 no.2
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    • pp.191-198
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    • 2023
  • As a member of the focal adhesion complex of the plasma membrane, Src is a nonreceptor tyrosine kinase that controls cell adhesion and motility. However, how Src activity is regulated in the plasma membrane microdomain in response to components of the extracellular matrix (ECM) remains unclear. This study compared and investigated the activity of Src in response to three representative ECM proteins: collagen type 1, fibronectin, and laminin. Genetically encoded FRET-based Src biosensors for plasma membrane subdomains were used. FRET-based biosensors allow the real-time analysis of protein activity in living cells based on their high spatiotemporal resolution. The results showed that Src activity was maintained at a high level under all ECM conditions of the lipid raft, and there was no significant difference between the ECM conditions. In contrast, Src activity was maintained at a low level in the non-lipid raft membrane. In addition, the Src activity of lipid rafts remained significantly higher than that of non-lipid raft regions under the same ECM conditions. In conclusion, this study demonstrates that Src activity can be controlled differently by lipid rafts and non-lipid raft microdomains.

Enhanced biological effects of Phe140Asn, a novel human granulocyte colony-stimulating factor mutant, on HL60 cells

  • Chung, Hee-Kyoung;Kim, Sung-Woo;Byun, Sung-June;Ko, Eun-Mi;Chung, Hak-Jae;Woo, Jae-Seok;Yoo, Jae-Gyu;Lee, Hwi-Cheul;Yang, Byoung-Chul;Kwon, Moo-Sik;Park, Soo-Bong;Park, Jin-Ki;Kim, Kyung-Woon
    • BMB Reports
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    • v.44 no.10
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    • pp.686-691
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    • 2011
  • Granulocyte colony-stimulating factor (G-CSF) is a cytokine secreted by stromal cells and plays a role in the differentiation of bone marrow stem cells and proliferation of neutrophils. Therefore, G-CSF is widely used to reduce the risk of serious infection in immunocompromised patients; however, its use in such patients is limited because of its non-persistent biological activity. We created an N-linked glycosylated form of this cytokine, hG-CSF (Phe140Asn), to assess its biological activity in the promyelocyte cell line HL60. Enhanced biological effects were identified by analyzing the JAK2/STAT3/survivin pathway in HL60 cells. In addition, mutant hG-CSF (Phe140Asn) was observed to have enhanced chemoattractant effects and improved differentiation efficiency in HL60 cells. These results suggest that the addition of N-linked glycosylation was successful in improving the biological activity of hG-CSF. Furthermore, the mutated product appears to be a feasible therapy for patients with neutropenia.

In vivo Antifungal Activity Against Various Plant Pathogenic Fungi of Curcuminoids Isolated from the Rhizomes of Curcuma longa

  • Cho, Jun-Young;Choi, Gyung-Ja;Lee, Seon-Woo;Lim, He-Kyoung;Jang, Kyung-Soo;Lim, Chi-Hwan;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.94-96
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    • 2006
  • In a search for plant extracts with potent in vivo antifungal activity against various plant pathogenic fungi, the methanol extract of the Curcuma longa rhizomes effectively controlled the development of rice blast catised by Magnaporthe grisea and tomato late blight caused by Phytophthora infestans. Three curcuminoids such as curcumin, demethoxycurcumin, and bisdemethoxycurcumin were purified from the methanol extract of C. longa rhizomes as antifungal principles. Among the three curcuminoids, demethoxycurcumin was the most active to both rice blast and tomato late blight, followed in order by curcumin and bisdemethoxycurcumin. However, they all exhibited no or little in vivo antifungal activity against other fungal pathogens causing rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), wheat leaf rust (Puccinia recondita), or barley powdery mildew (Blumeria graminis f. sp. hordel).

Development of Carbon-Based Solid Acid Catalysts Using a Lipid-Extracted Alga, Dunaliella tertiolecta, for Esterification

  • Ryu, Young-Jin;Kim, Z-Hun;Lee, Seul Gi;Yang, Ji-Hyun;Shin, Hee-Yong;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.732-738
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    • 2018
  • Novel carbon-based solid acid catalysts were synthesized through a sustainable route from lipid-extracted microalgal residue of Dunaliella tertiolecta, for biodiesel production. Two carbon-based solid acid catalysts were prepared by surface modification of bio-char with sulfuric acid ($H_2SO_4$) and sulfuryl chloride ($SO_2Cl_2$), respectively. The treated catalysts were characterized and their catalytic activities were evaluated by esterification of oleic acid. The esterification catalytic activity of the $SO_2Cl_2$-treated bio-char was higher ($11.5mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.\;^{-1}$) than that of commercial catalyst silica-supported Nafion SAC-13 ($2.3mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.^{-1}$) and $H_2SO_4$-treated bio-char ($5.7mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.^{-1}$). Reusability of the catalysts was examined. The catalytic activity of the $SO_2Cl_2$-modified catalyst was sustained from the second run after the initial activity dropped after the first run and kept the same activity until the fifth run. It was higher than that of first-used Nafion. These experimental results demonstrate that catalysts from lipid-extracted algae have great potential for the economic and environment-friendly production of biodiesel.

InhA-Like Protease Secreted by Bacillus sp. S17110 Inhabited in Turban Shell

  • Jung, Sang-Chul;Paik, Hyoung-Rok;Kim, Mi-Sun;Baik, Keun-Sik;Lee, Woo-Yiel;Seong, Chi-Nam;Choi, Sang-Ki
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.402-408
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    • 2007
  • A strain producing a potent protease was isolated from turban shell. The strain was identified as Bacillus sp. S17110 based on phylogenetic analysis. The enzyme was purified from culture supernatant of Bacillus sp. S17110 to homogeneity by ammonium sulfate precipitation, SP-Sepharose, and DEAE-Sepharose anion exchange chromatography. Protease activity of the purified protein against casein was found to be stable at pH 7 to pH 10 and around $50^{\circ}C$. Approximately 70% of proteolytic activity of the enzyme was detected either in the presence of 100 mM SDS or Tween 20. The enzyme activity was enhanced in the presence of $Ca^{2+},\;Zn^{2+},\;Mg^{2+}$, but was inhibited by EDTA, indicating that it requires metal for its activity. The purified enzyme was found to be a monomeric protein with a molecular mass of 75 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. The purified enzyme was analyzed through peptide fingerprint mass spectra generated from matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and a BLAST search, and identified as immune inhibitor A (inhA) deduced from nucleotide sequence of B. cereus G9241. Since InhA was identified as protease that cleave antibacterial proteins found in insect, inhA-like protease purified from Bacillus sp. S17110 might be pathogenic to sea invertebrates.

Novel Anticandidal Activity of a Recombinant Lampetra japonica RGD3 Protein

  • Wu, Caiping;Lu, Li;Zheng, Yuanyuan;Liu, Xin;Xiao, Rong;Wang, Jihong;Li, Qingwei
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.905-913
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    • 2014
  • Lj-RGD3, an RGD (Arg-Gly-Asp) toxin protein from the salivary gland of Lampetra japonica, exhibits antifungal activity against Candida albicans. Lj-RGD3 has three RGD motifs and shows homology to histidine-rich glycoprotein. We synthesised two mutant derivatives of Lj-RGD3: Lj-26, which lacks all three RGD motifs and contains no His residues; and Lj-112, which lacks only the three RGD motifs. We investigated the effects of the wild-type and mutated toxins on a gram-positive bacterium (Escherichia coli), a gram-negative bacterium (Staphylococcus aureus), and a fungus (C. albicans). rLj-RGD3 and its mutants exhibited antifungal but not antibacterial activity, as measured by a radial diffusion assay. The C. albicans inhibition zone induced by rLj-112 was larger than that induced by the other proteins, and its inhibitory effect on C. albicans was dose-dependent. In viable-count assays, the rLj-112 MIC was $7.7{\mu}M$, whereas the MIC of the positive control (ketoconazole) was $15{\mu}M$. Time-kill kinetics demonstrated that rLj-112 effectively killed C. albicans at $1{\times}$ and $2{\times}$ MIC within 12 and 6 h, respectively. Electron microscopy analysis showed that rLj-RGD3 and rLj-112 induced C. albicans lysis. Our results demonstrate a novel anticandidal activity for rLj-RGD3 and its mutant derivatives.

Isolation and Characterization of Calcite Forming Bacteria from Various Environments in Korea (다양한 환경에서의 탄산칼슘 생성 균주 분리 및 특성 연구)

  • Kim, YongGyeong;Kang, Chang-Ho;Oh, Soo Ji;So, Jae-Seong
    • KSBB Journal
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    • v.29 no.5
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    • pp.323-327
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    • 2014
  • Microbially induced calcite precipitation is a naturally occurring biological process in which microbes produce calcite on the surface of the microorganisms by urease activity. In order to collect calcite forming bacteria (CFB) in Korea, we isolated 343 putative CFB strains from various environments over three year period (2011~2013) and selected 100 CFB strains. Average of calcite productivity was 10.56 mg/mL. And average of ammonium concentration by urease activity was $8.00{\mu}M$. Two useful CFB strains of the others were analyzed by 16S rRNA and identified as Sporosarcina sp. and Viridibacillus arenosi. The CFB strains presented in this study are indigenous microorganisms in Korea and they are expected to be applicable to a variety of environments in the country.