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Diversity of Root-Associated Paenibacillus spp. in Winter Crops from the Southern Part of Korea

  • CHEONG HOON;PARK SOO-YOUNG;RYU CHOONG-MIN;KIM JIHYUN F.;PARK SEUNG-HWAN;PARK CHANG SEUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1286-1298
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    • 2005
  • The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.

Effect on the Production of Toxin Protein of Bacillus thuringiesis by Culture Conditions (배양 조건이 Bacillus thuringiesis 독소단백질의 생산에 미치는 영향)

  • Kim, Tae Wan;Kim, Tae Hwan;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.358-363
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    • 2014
  • We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

Comparative Proteome Analysis of Zerumbone-treated Helicobacter pylori (Zerumbone 처리에 따른 Helicobacter pylori의 단백질 비교분석)

  • Kim, Sa-Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.275-283
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    • 2018
  • Helicobacter pylori is a causative organism of various gastrointestinal diseases, including chronic gastritis, gastric ulcer, or gastric adenocarcinoma. Pathogenic factors, such as cytotoxin-associated protein A (CagA) and vacuolating cytotoxic protein A (VacA), play a role. This study analyzed qualitatively and quantitatively the effects of zerumbone on the changes in the protein expression levels of various H. pylori proteins, including CagA and VacA. Approximately 200 significant proteins were screened for the H. pylori 60190 (VacA positive / CagA positive; Eastern type) strain, and proteomic analysis was performed on 13 protein molecules that were clinically significant. After two-dimensional electrophoresis (2-DE), $ImageMaster^{TM}$ 2-DE Platinum software was used for quantitative measurements of protein spots. Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF-MS) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) were used for protein identification. After intensive analysis of the proteins that showed significant changes, a reverse transcription-polymerase chain reaction was performed as required to verify the results. In this study, the significance of zerumbone as a therapeutic agent for H. pylori infection was examined by screening a new pharmacological activity mechanism of zerumbone.

Antimicrobial Activities of Propolis against Oral Candidiasis by Candida Albicans -Effect of Microbial Inhibition Using Propolis- (구강 캔디다증 알비칸스에 대한 프로 폴리스의 항균 활동 -프로폴리스를 이용한 미생물 억제 효과-)

  • Kwun, Hyeon-Sook;Nam, Seoul-Hee;Park, Min-Kyoung;Cho, Mi SooK;Cheon, Sae-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.9
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    • pp.5644-5651
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    • 2014
  • Propolis is an extremely safe natural antimicrobial substance that has been reported to have powerful antibacterial efficacy. The aim of this study was to evaluate the inhibitory effects of propolis against Candida albicans (C. albicans). Propolis was collected from the honey bee Apis mellifera. The strain of C. albicans was cultivated overnight in liquid media incubated at $37^{\circ}C$. The antimicrobial activity was investigated using phosphate buffered saline (PBS), 3% sodium hypochlorite (NaOCl), 0.1% chorhexidine (CHX), and propolis extracts ($5{\mu}l/ml$, $10{\mu}l/ml$). C. albicans were sensitive to 3% NaOCl, 0.1% CHX, and propolis ($5{\mu}l/ml$, $10{\mu}l/ml$) with zones of inhibition of 15, 14.5, 16, and 17 mm, respectively. The CFU of PBS, 3% NaOCl, 0.1% CHX, $5{\mu}l/ml$ and $10{\mu}l/ml$ of propolis led a 1, 7, 7, 5 and 7-log reduction. Among the groups tested, C. albicans was most sensitive to $10{\mu}l/ml$ of propolis, which showed the largest inhibition zones. Therefore, propolis can be a new antimicrobial therapy for oral mucosa disease in traditional medicine.

The Site-Directed A184S Mutation in the HTH Domain of the Global Regulator IrrE Enhances Deinococcus radiodurans R1 Tolerance to UV Radiation and MMC Shock

  • Zhang, Chen;Zhou, Zhengfu;Zhang, Wei;Chen, Zhen;Song, Yuan;Lu, Wei;Lin, Min;Chen, Ming
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2125-2134
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    • 2015
  • IrrE is a highly conserved global regulator in the Deinococcus genus and contributes to survival from high doses of UV radiation, ionizing radiation, and desiccation. Drad-IrrE and Dgob-IrrE from Deinococcus radiodurans and Deinococcus gobiensis I-0 each share 66% sequence identity. However, Dgob-IrrE showed a stronger protection phenotype against UV radiation than Drad-IrrE in the D. radiodurans irrE-deletion mutant (ΔirrE), which may be due to amino acid residues differences around the DNA-binding HTH domain. Site-directed mutagenesis was used to generate a Drad-IrrE A184S single mutant, which has been characterized and compared with the ΔirrE mutant complemented strain with Drad-irrE, designated ΔirrE-E. The effects of the A184S mutation following UV radiation and mitomycin C (MMC) shock were determined. The A184S mutant displayed significantly increased resistance to UV radiation and MMC shock. The corresponding A184 site in Dgob-IrrE was inversely mutated, generating the S131A mutant, which exhibited a loss of resistance against UV radiation, MMC shock, and desiccation. qPCR analysis revealed that critical genes in the DNA repair system, such as recA, pprA, uvrA, and ddrB, were remarkably induced after UV radiation and MMC shock in the ΔirrE-IE and A184S mutants. These data suggested that A184S improves the ability against UV radiation and MMC shock, providing new insights into the modification of IrrE. We speculated that the serine residue may determine the efficiency of DNA binding, leading to the increased expression of IrrE-dependent genes important for protection against DNA damage.

Cloning, Expression, and Characterization of a New Xylanase from Alkalophilic Paenibacillus sp. 12-11

  • Zhao, Yanyu;Meng, Kun;Luo, Huiying;Yang, Peilong;Shi, Pengjun;Huang, Huoqing;Bai, Yingguo;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.861-868
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    • 2011
  • A xylanase gene, xyn7c, was cloned from Paenibacillus sp. 12-11, an alkalophilic strain isolated from the alkaline wastewater sludge of a paper mill, and expressed in Escherichia coli. The full-length gene consists of 1,296 bp and encodes a mature protein of 400 residues (excluding the putative signal peptide) that belongs to the glycoside hydrolase family 10. The optimal pH of the purified recombinant XYN7C was found to be 8.0, and the enzyme had good pH adaptability at 6.5-8.5 and stability over a broad pH range of 5.0-11.0. XYN7C exhibited maximum activity at $55^{\circ}C$ and was thermostable at $50^{\circ}C$ and below. Using wheat arabinoxylan as the substrate, XYN7C had a high specific activity of 1,886 U/mg, and the apparent $K_m$ and $V_{max}$ values were 1.18 mg/ml and 1,961 ${\mu}mol$/mg/min, respectively. XYN7C also had substrate specificity towards various xylans, and was highly resistant to neutral proteases. The main hydrolysis products of xylans were xylose and xylobiose. These properties make XYN7C a promising candidate to be used in biobleaching, baking, and cotton scouring processes.

Cloning, Heterologous Expression, and Characterization of Novel Protease-Resistant ${\alpha}$-Galactosidase from New Sphingomonas Strain

  • Zhou, Junpei;Dong, Yanyan;Li, Junjun;Zhang, Rui;Tang, Xianghua;Mu, Yuelin;Xu, Bo;Wu, Qian;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1532-1539
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    • 2012
  • The ${\alpha}$-galactosidase-coding gene agaAJB13 was cloned from Sphingomonas sp. JB13 showing 16S rDNA (1,343 bp) identities of ${\leq}97.2%$ with other identified Sphingomonas strains. agaAJB13 (2,217 bp; 64.9% GC content) encodes a 738-residue polypeptide (AgaAJB13) with a calculated mass of 82.3 kDa. AgaAJB13 showed the highest identity of 61.4% with the putative glycosyl hydrolase family 36 ${\alpha}$-galactosidase from Granulicella mallensis MP5ACTX8 (EFI56085). AgaAJB13 also showed <37% identities with reported protease-resistant or Sphingomonas ${\alpha}$-galactosidases. A sequence analysis revealed different catalytic motifs between reported Sphingomonas ${\alpha}$-galactosidases (KXD and RXXXD) and AgaAJB13 (KWD and SDXXDXXXR). Recombinant AgaAJB13 (rAgaAJB13) was expressed in Escherichia coli BL21 (DE3). The purified rAgaAJB13 was characterized using p-nitrophenyl-${\alpha}$-D-galactopyranoside as the substrate and showed an apparent optimum at pH 5.0 and $60^{\circ}C$ and strong resistance to trypsin and proteinase K digestion. Compared with reported proteaseresistant ${\alpha}$-galactosidases showing thermolability at $50^{\circ}C$ or $60^{\circ}C$ and specific activities of <71 U/mg with or without protease treatments, rAgaAJB13 exhibited a better thermal stability (half-life of >60 min at $60^{\circ}C$) and higher specific activities (225.0-256.5 U/mg). These sequence and enzymatic properties suggest AgaAJB13 is the first identified and characterized Sphingomonas ${\alpha}$-galactosidase, and shows novel protease resistance with a potential value for basic research and industrial applications.

Reinforcement of Collapsed Railway Subgrade and Line Capacity Increase Using Short Reinforcement with Rigid Wall (짧은 보강재와 일체형 강성벽체를 활용한 철도 붕괴노반 보강 및 선로용량 증대 기술)

  • Kim, Dae-Sang
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.11
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    • pp.604-609
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    • 2016
  • This study evaluated the long-term performance of RSR (Reinforced Subgrade for Railways) technology which increases the railway line capacity without the need for additional land. Its characteristics include the use of a short reinforcement with rigid wall, which make it possible to apply it in confined spaces. The 7m high and 40m long testbed employed to evaluate the long-term performance was designed and constructed near Jupo station on the Chang-hang line. This line, located close to a local bus route, had collapsed at the subgrade following heavy rainfall. The performance of the new type of subgrade was verified with long term measurements over a 2 year period including the surface and ground settlement, horizontal displacement of the wall, tensile strain of the reinforcement, and settlement of the rail top on the side track. Based on the results of the measurements made until now, we concluded that it had sufficient safety and serviceability for use as a railway subgrade. It is expected that RSR technology could be frequently used at sites which lack the necessary construction materials for an embankment and are located close to functional railway lines and boundaries, in order to settle civil complaints.

A Study on Traditional Korean Medical Countermeasures of Influenza A by Case Histories (의안(醫案)을 통한 신종 인플루엔자의 한의학적 대처방안)

  • Ahn, Sang-Young;Han, Chang-Hyun;Kwon, Oh-Min;Park, Sang-Young;Ahn, Sang-Woo
    • Journal of Korean Medical classics
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    • v.22 no.4
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    • pp.221-239
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    • 2009
  • Background: The recent outbreak of the novel strain of influenza A (H1N1) virus has raised a global concern of the future risk of pandemic. Traditional Korean Medicine(TKM) has been combatting against contagious diseases and developed its own particular and efficient way in treating those diseases. Objectives: Provide a distinctive and effective TKM method in understanding the principles of treatment, prevention, and contraindications against influenza A through case histories. Method: We revised case histories of eminent doctors of Ming and Qing dynasties according to their clinical manifestations similar to those of influenza A. We also verified prescriptions of the "Dong-uibogam(東醫寶鑑)" through examining clinical practices of China today. Results: 1. The subtypes of Warm disease; Wind Warmth and Pestilence has the most similar clinical manifestation in comparison to Influenza A. Specifying these terms is important in recognizing and classifying various diseases under the name of Warm disease. 2. TKM considered not only external factors but also individual factors like general condition, diet, emotion, constitutional types, etc in the treatment of febrile diseases. 3. TKM developed a new way in the treatment of contagious diseases. considering the characteristics of each pathogen. TKM described the Warm pathogen being light as a feather, which enters through the nose, principally affecting the Lung. Therefore, they emphasized treatment with Pungent Sweet Cooling Moistening[辛甘凉潤] method. 4. As the conservation of fluid and humor is the primordial concern in the treatment of Warm disease, they restricted release of the exterior with pungent-warm and purgation method. The purgation methods was used exclusively to decrease fever and preserve fluid and humor. 5. The only differentiating characteristics of Influenza A with seasonal influenza are vomiting and diarrhea. Case research revealed the possibility of these manifestations to be a mechanism of restoration. 6. TKM provides alimento prevention method like Mint Pear Porridge, mung bean, and etc also combination of herbal medicine. Also emphasized in the conservation of essence for the prevention of contagious diseases. Conclusions: TKM developed its unique way in understanding the pathogenesis, diagnosis, treatment, and prevention of contagious diseases and formed its independent scheme of Warm Disease. This knowledge in febrile contagious disease is relevant today in providing diverse treatment and prevention for influenza A.

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Improvement of Mechanical Properties of Mg alloys through Control of Grain Size and Texture (결정립크기와 집합조직제어를 통한 마그네슘 합금의 기계적 성질 개선)

  • Kim, W.J.;Lee, J.B.;Kim, W.Y.;Jeong, H.G.;Park, J.D.
    • Proceedings of the Korean Society for Technology of Plasticity Conference
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    • 2006.05a
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    • pp.57-58
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    • 2006
  • The effects of lowering ECAP temperature during ECAP process and Post-ECAP annealing on microstructure, texture and mechanical properties of the AZ31 alloys have been investigated in the present study. The as-extruded materials were ECAP processed to 2 passes at 553K prior to subsequent pressing up to 6 passes at 523K or 493K. When this method of lowering ECAP temperature during ECAP was used, the rods could be successfully deformed up to 6 passes without any surface cracking. Grain refinement during ECAP process at 553K might have helped the material to endure further straining at lower deformation temperatures probably by increasing the strain accommodation effect by grain boundary sliding, causing stress relaxation. Texture modification during ECAP has a great influence on the strength of Mg alloys because HCP metals have limited number of slip systems. As slip is most prone to take place on basal planes in Mg at room temperature, the rotation of high fraction of basal planes to the directions favorable for slip as in ECAP decreases the yield stress appreciably. The strength of AZ31 Mg alloys increases with decrease of grain size if the texture is constant though ECAP deformation history is different. A standard positive strength dependence on the grain size for Mg alloys with the similar texture (Fig. 1) supports that the softening of ECAPed Mg alloys (a negative slope) typically observed despite the significant grain refinement is due to the texture modification where the rotation of basal planes occurs towards the orientation for easier slip. It could be predicted that if the original fiber texture is restored after ECAP treatment yielding marked grain refinement, yield stress as high as 500 MPa will be obtained at the grain size of ${\sim}1{\mu}m$. Differential speed rolling (DSR) with a high speed ratio between the upper and lower rolls was applied to alter the microstructure and texture of the AZ31 sheets. Significant grain refinement took place during the rolling owing to introduction of large shear deformation. Grain size as small as $1.4{\mu}m$ could be obtained at 423K after DSR. There was a good correlation between the (0002) pole intensity and tensile elongation. This result indicates that tensile ductility improvement in the asymmetrically rolled AZ31 Mg alloys is closely related to the weakening of basal texture during DSR. Further basal texture weakening occurred during annealing after DSR. According to Hall-Petch relation shown in Fig. 1, the strength of the asymmetrically rolled AZ31 is lower than that of the symmetrically rolled one when compared at the same grain size. This result was attributed to weakening of fiber texture during DSR. The DSRed AZ31, however, shows higher strength than the ECAPed AZ31 where texture has been completely replaced by a new texture associated with high Schmid factors.

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