J_Environmetal Microbiology and Engineering

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J-36 Simultaneous Quantification of Cyanobacteria and Microcystis spp. using Quantitative Realtime PCR Kyoung-Hee OH, Seung-Hee SHIN, Mi-Na YU, Eun-Hye PARK and Young-Cheol CHO* Dept. of Environmental Engineering, Chungbuk National University, Cheongju 361-763, Korea. *Corresponding author: choy@cbnu.ac.kr Microcystins are produced mainly by Microcystis sp. at large reservoirs in Korea. Therefore, quantification of Microcystis is required to monitor the potential for algal-toxin production. It is difficult to count Microcystis, however, because these mostly form scum. In this study, new method was developed to quantify cyanobacteria and Microcystis simultaneously using quantitative realtime PCR. The primers and probe, cyanobacteriaspecific and Microcystis-specific primers and Microcystis-specific TaqMan probe, were designed from the conserved regions of 16S ribosomal RNA gene sequences for cyanobacteria and Microcystis, respectively. It was proved as an adequate and reproducible method to quantify cyanobacteria and Microcystis in the environmental samples. It is expected that the use of this method enhances the accuracy of Microcystis counts and reduces the time involved in the analysis of samples by microscopy. Also, this method enables us to monitor the formation of cyanobacterial mass occurrences in situ and to reveal the factors promoting the growth of Microcystis. [Supported by grants from National Science Foundation of Korea] Keywords: Microcystis, Real-time PCR, Simultaneous quantification J-37 GFP Tagging of p-Cresol Methylhydroxylase in Pseudomonas alkylphenolia and Its Dominant Expression in Colonies CHO AH RA and Kyoung LEE* Dept. of Microbiology, Changwon National University, Kyongnam 641-773, Korea. *Corresponding author: 0506joara@hanmail.net In this study, the chromosome-encoded pcuRCAXB genes that are required for p-cresol degradation have been identified by using the newly constructed Green fluorescence protein (GFP)-based promoter probe transposon in the long-chain alkylphenol degrader, Pseudomonas alkylphenolia. The deduced amino acid sequences of the genes showed the highest identities at the levels of 65-93% compared to those in the databases. The transposon was identified to be inserted in the pcuA gene with the promoterless gfp gene being under the control of the pcu catabolic gene promoter. The expression of GFP was positively induced by p-cresol and was about 10 times higher by cells grown on agar than those in liquid culture. This result indicates that P. alkylphenolia additionally possesses a protocatechuate ortho cleavage route for p-cresol degradation that is dominantly expressed in colonies. Keywords: Pseudomonas, transposon 2011 International Symposium & Annual Meeting Isolation and Characteristics of Entomopathogenic Fungi Beauveria bassiana as Insectide Agent Against Greenhouse Ahitefly(Trialeurodes vaporariorum) Chang-Su KIM 2 , Keun-Hyung LEE 3 , Young-Ho NAM 3 , Gyeong-Muk KANG 3 , Jin-Won KIM 4 and Gi-Seok KWON* 1 1 Departmentl of bioresource Sciences Andong Nation University, Andong 760-749, Korea. 2 안동대학교. 3 School of bioresource Sciences, Andong Nation University, Andong 760-749, Korea. 4 J-38 Yecheon Agricuture Technology Center, Yecheon 757-802, Korea. *Corresponding author: ckdtn0629@hanmail.net The object of this study is to use Entomopathogenic fungi B.bassiana as agent Greenhouse whitefly(Trialeaarodes vaporariorum). Each 4 strains were prepared as follows : B.bassiana was parceled by Rural Development Administration(J157&J216), Gyougbuk Agricutural Reserch&Extension Service(S186) Yecheon Agricultuer Technology Center(B.K). Chitinase, protease, lipase are important and involved in the host invasion. In vitro, plate assay was used to observe the each enzyme. And chitinase was observed by three strains. Also, Protease, lipase was observed by all strains. And activity measurement of enzyme is in progress. In vivo, one control plot and each four treatment plots was treated to 50 Greenhouse whiteflys in a capsicum. After a few days, B.bassiana spore(60g/1L) that was mixtuerd the yolk of an egg spraied due volta 500ml in each treatment plots and spraied a Neoseiulus californicus in the control plot. Later, indicated Greenhouse whitefly that enticed yellow flypaper trap and compared control plot with treatment plots. Consequently, B.bassiana was a predominant and confirmed as insecticide agent against Greenhouse whitefly.[ This study supported by Technology of Development program for Agriculture and Foresty, Gyeongbuk 2011] Keywords: Beauveria bassiana, Entomopathogenic, Greenhouse whitefly J-39 Effect of Nitrate on the Electricity Generation of Microbial Fuel cell Jae Kyung JANG*, Jung Eun CHOI, Young Sun RYOU, Sung Hyoun LEE, Jong Goo KIM, Youn Koo KANG, Young Hwa KIM and Hyung Mo LEE National Academy of Agricutural Science. *Corresponding author: jkjang1052@korea.kr The microbial fuel cells (MFCs) can convert to useful electricity from artificial wastewater including nitrogen source such as nitrate (NO3 - ) and also to treat this wastewater. Electrons and protons produced by metabolic process of microorganisms in the anode compartment transfer to cathode through the out circuit and membrane, respectively. However, when the nitrate is included in the wastewater (fuel), it can act the electron donor. It will decrease the current value. These studies were conducted to investigate the influence of nitrate concentration on the current generation. Sodium acetate was used as electron donor and the chemical oxygen demand (CODcr) of the wastewater was around 250 mg L -1 . The microbial fuel cells produced a current about 9 mA stably when 249.2±4.6 mg/L artificial wastewater as COD including only acetate without nitrate was fed. When nitrate from 31.4±0.4 to 123.3±0.1 mg/L was fed into MFCs with sodium acetate, the current generation decreased from 9.97±0.06 to 0.20±0.01 mA according to increase the nitrate concentration. However nitrate concentration was removed up to 96% as added nitrate of 123.3±0.1 mg/L. These results showed that nitrate could behave the electron acceptor in the anode compartment. Keywords: Microbial fuel cell, nitrate, electricity generation The Korean Society for Microbiology and Biotechnology 405
Bioelectrochemical Fixation of CO2 Bo Young JEON 1 , Ji-Won CHOI 1 , Il Lae JUNG 2 and Doo Hyun PARK* 1 1 Department of Chemical and Biological Engineering, Seokyeong University, Seoul 136-704, Korea. 2 J-40 Department of Radiation Biology, Environmental Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon 305-353, Korea. *Corresponding author: baakdoo@skuniv.ac.kr For the isolation of CO2-fixing bacteria using electrochemical reducing power (ERP), a plate-type electrochemical bioreactor (PEB) was employed. Soluble neutral red (NR) was used as an electron mediator for carbonate-basal agar medium (CBAM) prepared in the PEB. For test of bacterial CO2 fixation using the ERP, a single-compartmented electrochemical bioreactor (SCEB) was employed. NR immobilized in the graphite felt cathode (NR-cathode) was used as an electron mediator for carbonate-basal broth medium (CBBM) prepared in the SCEB. Two bacterial genera capable of electrochemically fixing CO2 were isolated using the PEB and cultivated using the SCEB. The isolated bacterial species were 99% identified with Achromobacter sp. and Alcaligenes sp. Approximate 150 ml of CO2 per day was consumed by 300 ml of the mixed culture of Achromobacter sp. and Alcaligenes sp. grown electrochemically in the SCEB. Acknowledgement This work was supported by the New &Renewable Energy of the Korea Institute of Energy Technology Evaluation and Planning (KETEP) grant funded by the Korea governmental Ministry of Knowledge Economy (2010T1001100334) Keywords: CO2-fixing bacteria, neutral red, electrochemical reducing power, Alcaligenes sp., Achromobacter sp. Inactivation of Chloramphenicol and Florfenicol by a Novel Chloramphenicol Hydrolase from Alluvial Soil Metagenome Weixin TAO 1 , Myung Hwan LEE 2 , Jing WU 2 , Nam Hee KIM 2 , Jin-Cheol KIM 3 , Eul Chul HWANG 2 and Seon-Woo LEE* 1,2 1 Department of Medical Bioscience, Dong-A University, Busan 604-714, South Korea. 2 Department of Applied Biology, Dong-A University, Busan 604-714, South Korea. 3 J-41 Chemical Biotechnology Research Center, KRICT, Daejeon 305-343, South Korea. *Corresponding author: Seonlee@dau.ac.kr In a previous study, EstDL136 isolated from alluvial soil metagenome was redefined as chloramphenicol (Cml) acetate esterase, on account of its capability of Cml reactivation that against Cml acetyltransferase (CAT). Further investigation of Cml catalytic activity in the absence of CAT revealed that Cml was metabolized by EstDL136. The metabolite was identified as p-nitrophenylserinol by LC-MS and 1 H-NMR, suggesting a promiscuous amidase activity of the enzyme. Purified EstDL136 catalyzed Cml hydrolysis when they were co-incubated. Residues were detected by HPLC, which showed that Cml was gradually hydrolyzed and the hydrolysate, p-nitrophenylserinol was concurrently generated. EstDL136 also hydrolyzed florfenicol (Ffl), the synthetic florinated analog of Cml that was resistant to CAT. Therefore, Escherichia coli was tolerant to Cml or Ffl when estDL136 was expressed, and duration of the lag phase was highly correlated with the concentrations of supplemented antibiotics (Cml, Ffl), indicating that estDL136 mediated the amphenicols resistance. This study primarily reported a novel bacterial Cml hydrolase that also hydrolyzes Ffl, conferring Cml and Ffl resistance to the E. coli. Keywords: Chloramphenicol, Florfenicol, Hydrolase 406 www.kormb.or.kr J-42 Efficacy of Combined Application of Entomopathogenic Fungal Conidia and Botanical Insecticides for Aphid Control Tuan anh PHAM and Keun KIM* Dept. of Bioscience and Biotechnology, Univ. of Suwon, Gyeonggi-do 445-743, Korea. *Corresponding author: ptanh140380@yahoo.com Some botanical insecticides prepared from neem, derris, garlic and chinaberry were evaluated for the influence of the botanical insecticides on susceptibility of conidia of Beauveria bassiana by the germination ratio of conidia. The results demonstrated that botanical insecticides significantly reduced the germination ratio of conidia and it could not be mixed with the conidia in oil-based formulation for long-term storage. After that, the virulence of oil-based formulation of conidia in combination with botanical insecticides against aphid Myzus persicae was examined with detached leaf test. The botanical insecticide was mixed with the oil-formulated conidia just before spraying and the result showed that the combined application of two insecticides significantly increased the mortality of the aphid. Keywords: Beauveria bassiana conidia, botanical insecticides, aphid control Seasonal Escherichia coli Genotypes Determined by Horizontal Fluorophore-Enhanced rep-PCR (HFERP) in the Yeongsan River Basin of South Korea Jeonghwan JANG 1 , Tatsuya UNNO 1 , Seung Won LEE 1 , Kyung Hwa CHO 1 , Michael J. SADOWSKY 3,4 , GwangPyo KO 5 , Joon Ha KIM 1 and Hor-Gil HUR* 1,2 1 School of Environmental Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea. 2 International Environmental Research Center, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea. 3 Department of Soil, Water and Climate, University of Minnesota, St. Paul, Minnesota, USA. 4 BioTechnology Institute, University of Minnesota, St. Paul, Minnesota, USA. 5 J-43 Department of Environmental Public Health, Seoul National University, Republic of Korea. *Corresponding author: hghur@gist.ac.kr Seasonal and spatial variation in the genotypic diversity of 3,480 Escherichia coli isolates obtained from the Yeongsan River basin in South Korea was investigated by using the horizontal fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting technique. The MDS analysis, done based on HFERP DNA fingerprints, showed that E. coli isolates obtained in October through December clustered tightly, while those obtained in other sampling periods were more genetically diverse. SOMs analysis, done using the 10 most frequently-isolated E. coli genotypes, showed the occurrence of season-specific E. coli genotypes and the main SOMs clusters were most influenced by temperature, strain diversity, and biochemical oxygen demand. Diversity among E. coli genotypes tended to decrease as water temperature decreased, and the numbers of E. coli genotypes observed in urban area were greater, more diverse, and less dependent on water temperature than those obtained from agricultural areas. This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (MEST) (NRF-R1A5A004-2010-0029224), and also by the Korea Environmental Technology &Industry Institute (KEITI) as the Next Generation Eco Innovation Technology Development Project. Keywords: Escherichia coli, Yeongsan River, Horizontal Fluorophore-Enhanced rep-PCR
Page 1 and 2: Isolation and Characterization of L
Page 3 and 4: Characterization of Phosphate-Solub
Page 5 and 6: Thermodynamic and Kinetic Analyses
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J_Environmetal Microbiology and Engineering

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