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이영득,이지현,심원보,Don Anushka Sandaruwan Elvitigala,디조이사마하나마,이수진,허수진,이제희,강도형,오철홍 한국해양과학기술원 2014 Ocean science journal Vol.49 No.4
Glucanases are involved in degradation of glucans. Here, we report a new endo-β-1,3-glucanase Mzl86 identified in Mesoflavibacter zeaxanthinifaciens S86. The deduced amino-acid sequence of Mzl86 showed highest similarity (45.1%) with Leeuwenhoekiella blandensi and thus placed in glycosyl hydrolase family 16. Purified recombinant protein (rMz186) showed an optimum enzyme activity against laminarin at 50°C and pH 8. The enzyme was stable at 50°C for 1 hour (maintaining 80% of its maximum activity) and was strongly activated (187%) in the presence of 2.5 mM manganese. Substrate-specific activities of rMzl86 against laminarin, barley β-glucan and lichenan were 261, 128 and 115 unit/mg, respectively. rMzl86 degraded laminarioligosaccharides (lager than biose) and laminarin while producing mainly biose and glucose. Molecular and biochemical properties reveal that rMzl86 shares typical features of β-1,3-glucanase (EC 3.2.1.39) and thus is a potential candidate for use in agriculture, drug, chemical and bioethanol industries.
Dananjaya S.H.S.,GODAHEWAGELSHAN IMARSHANA,이영득,조종기,이제희,디조이사마하나마 한국임상수의학회 2014 한국임상수의학회지 Vol.31 No.6
Recently nano particles have proven for wide array of bioactive properties. In the present study, antibacterialproperties of chitosan silver nano composites (CAgNCs) were investigated against fish pathogenic Edwardsiella tarda. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs against E. tarda were 25 μg/mL and 125 μg/mL, respectively. The field emission scanning electron microscope (FE-SEM) imageof CAgNCs treated E. tarda showed the strongly damaged bacteria cells than non-treated bacteria. Furthermore,treatment of CAgNCs induced the level of intracellular reactive oxygen species (ROS) in E. tarda cells in concentrationand time dependent manner suggesting that it may generate oxidative stress leading to bacterial cell death. In addition,MTT assay results showed that the lowest cell viability at 100 μg/mL of CAgNCs treated E. tarda. Overall resultsof this study suggest that CAgNCs is a potential antibacterial agent to control pathogenic bacteria.
Udeni Menike,Krishan Ariyasiri,최진영,이영덕,W.D.N. Wickramaarachchi,H.K.A. Premachandra,디조이사마하나마,이지희 한국패류학회 2013 The Korean Journal of Malacology Vol.29 No.2
Cathepsins are lysosomal /cysteine proteases belong to papain family (C1 family) that is involved in intracellular protein degradation, antigen processing, hormone maturation, and immune responses. In this study, member of cathepsin family was identified from Manila clam (Mc-Cathepsin D) and investigated the immune response against brown ring disease (BRD) causing Vibrio tapetis challenge. The identified Mc-Cathepsin D gene encodes characteristic features typical for the cathepsin family including eukaryotic and viral aspartyl protease signature domain and two highly conserved active sites (84VVFDTGSSNLWV95 and 270IADTGTSLLAG281). Moreover, MC-Cathepsin D shows higher identity values (-50-70%) and conserved amino acids with known cathepsin D members. Transcriptional results (by quantitative real-time RT-PCR) showed that Mc-Cathepsin D was expressed at higher levels in gills and hemocytes than mantle, adductor muscle, foot, and siphon. After the V. tapetis challenge under laboratory conditions, Mc-Cathepsin D mRNA was up-regulated in gills and hemocytes. Present study indicates that Mc-Cathepsin D is constitutively expressed in different tissues and potentially inducible when infecting BRD by V. tapetis. It is further suggesting that Mc-Cathepsin D may be involved in multiple role including immune response reactions against BRD.
서승범,신상엽,S. H. S. Dananjaya,A. B. K. H. De Silva,니카피티야차밀라니,조종기,박건후,오철홍,강도형,디조이사마하나마 韓國臨床獸醫學會 2017 한국임상수의학회지 Vol.34 No.4
The aim of this study was to investigate the antibacterial properties of chitosan silver nanocomposites (CAgNCs) using pathogenic Vibrio ichthyoenteri as a bacterial model. Results of agar disc diffusion and turbidimetric assays showed that CAgNCs could inhibit the growth of V. ichthyoenteri in concentration dependent manner. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs were 75 and 125 μg/mL, respectively. Furthermore, CAgNCs treatment induced the reactive oxygen species (ROS) level in V. ichthyoenteri cells in concentration and time dependent manner, suggesting that it generates oxidative stress, leading to bacterial cell death. The field emission scanning electron microscope (FE-SEM) images of CAgNCs treated V. ichthyoenteri exhibited strong cell membrane damage than un-treated control bacteria. MTT assay results showed the highest cell viability (22%) at 75 μg/mL of CAgNCs treated bacteria samples. The results from this study suggest that CAgNCs is a potential antibacterial agent to control fish pathogenic bacteria.
Chang-Ning Li,Manoj-Kumar Srivastava,Qian Nong,Li-Tao Yang,Yang-Rui Li,디조이사마하나마 한국유전학회 2013 Genes & Genomics Vol.35 No.1
Abscisic acid (ABA) plays important roles in adaptive responses to various environmental stresses. The rate-limiting step in ABA biosynthesis is the oxidative cleavage of cis-epoxycarotenoids, which is catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED). In this experiment,a full-length cDNA encoding NCED gene was cloned by RT-PCR and RACE from sugarcane (Saccharum officinarum L.). The full-length of SoNCED is 2,521 bp with 1,827 bp open reading frame, encoding a peptide of 608amino acids. The calculated molecular weight of protein was 65.9 kDa with isoelectric point of 6.04. Conserved domains prediction indicated a chloroplast-targeting peptide located at N-terminus of SoNCED. Phylogenetic tree, constructed by Neighbor-Joining method indicated that SoNCED shared high identity with the NCEDs reported from other plant species. Sequence alignment revealed that the basic secondary structure including a-helices, b-strands, b-propeller and His residues coordinating catalytic sites of SoNCED were highly conserved as in the NCEDs from other plants. Tissue specific expression analysis using quantitative real-time PCR showed a significant increase in SoNCEDmRNAlevel and its correlation with O2– production rate and ABA accumulation in leaves and roots of sugarcane variety GT21 when exposed to water stress. Further, the stimulation of SoNCED mRNA level, O2– production rate and ABA content after exogenous application of ABA (100 lMol l-1) proved its involvement in pathways providing tolerance to drought stress.