J_Environmetal Microbiology and Engineering
J_Environmetal Microbiology and Engineering
J_Environmetal Microbiology and Engineering
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Isolation <strong>and</strong> Characterization of Lactobacillus Pentosus Kimchi<br />
from the Fermented Korean Food, Cutlassfish Kimchi<br />
Ji-Hyeong KIM 1 , Mi-Sun KWAK 1 , Hyeon-Jun SON 1 , Se-Jeung JANG 2 ,<br />
Jong-Bok RHO 3 , Young-Ki CHOI 4 <strong>and</strong> Moon-Hee SUNG* 1<br />
1<br />
Dept. of Advanced Fermentation Fusion Science &Technology, Kookmin<br />
University, Korea. 2 Dept. Bio &Nanochemistry, Kookmin University, Seoul,<br />
Korea. 3 BioLeaders Corporation, Daejeon, Republic of Korea. 4 I-101<br />
College of<br />
Medicine &Medical Research Institute, Chungbuk National University, Korea.<br />
*Corresponding author: safe2112@hanmail.net<br />
Kimchi, a Korean traditional fermented food, is prepared with various<br />
vegetables such as cabbage, radishes, spices, <strong>and</strong> other seasonings. The<br />
fermentation of kimchi is carried out by microorganisms, mainly GRAS<br />
lactic acid bacteria (GRAS-LAB). Recently, several health functions of<br />
kimchi have been reported, including antimicrobial effects <strong>and</strong> anti-viral<br />
effects. In order to isolate kimchi LAB, which has antimicrobial <strong>and</strong><br />
anti-viral effects, we screened the kimchi LAB from cutlassfish kimchi.<br />
We isolated kimchi LAB that could grow in an MRS plate medium<br />
containing 2% CaCO3. Finally, one strain was selected for the protease<br />
activity on a 1% milk plate medium. The isolated strain was identified<br />
according to its phenotypic characteristics <strong>and</strong> the data from the<br />
phylogenetic sequence analysis of the 16S rRNA gene. We examined<br />
antimicrobial activity, antifungal activity, <strong>and</strong> anti-viral activity with<br />
cell-free supernatants <strong>and</strong> a culture broth of kimchi LAB grown in MRS<br />
broth. [This work was supported by the Seoul R&BD Program (10580)<br />
<strong>and</strong> Top Br<strong>and</strong> Project grant from Korea Research Council of Fundamental<br />
Science &Technology (KGM3110912).]<br />
Keywords: fermentation, Kimchi, lactobacillus<br />
J _ <strong>Environmetal</strong> <strong>Microbiology</strong> <strong>and</strong> <strong>Engineering</strong><br />
J-1<br />
Growth Promoting Effect of Alginate on Nannochloropsis oculata<br />
Hyun-Jin PARK 1 , Young-Hwa KIM 2 <strong>and</strong> Jae-Hwa LEE* 1<br />
1 Department of Bioscience <strong>and</strong> Biotechnology, College of Medical <strong>and</strong> Life<br />
Science, Silla University, Busan 617-736, Korea. 2 Department of Pharmaceutical<br />
<strong>Engineering</strong>, College of Medical <strong>and</strong> Life Science, Silla University, Busan<br />
617-736, Korea.<br />
*Corresponding author: phj0386@naver.com<br />
The growth of Nannochloropsis oculata (N. oculata), unicellular<br />
microalgae, by alginate was investigated. Alginate was depolymerized with<br />
sulfuric acid (H2SO4) <strong>and</strong> heat (121℃), simultaneously. Addition of 0.75%<br />
alginate oligomer depolymerized with 0.2 N H2SO4 showed the maximum<br />
yield <strong>and</strong> the growth rate of N. oculata. Chlorophyll content <strong>and</strong> reducing<br />
sugar was increased by alginate oligomer in a dose-dependent manner.<br />
Alginate oligomer promoted the growth of N. oculata, whereas the original<br />
alginate polysaccharides had no significant effect. Laminaria japonica (L.<br />
japonica) extract containing high level of alginate was also increased<br />
growth rate <strong>and</strong> chlorophyll content. CO2 supply addition to L. japonica<br />
extract showed no change the growth rate, although addition to alginate<br />
oligomer showed prominently increased. N. oculata could use more<br />
saccharides in presence of CO2 according to reducing sugar determination.<br />
From these results, it is useful to establish optimal condition for high<br />
cell density cultivation of N. oculata.<br />
Keywords: Alginate, Nannochloropsis oculata, Laminaria japonica<br />
396 www.kormb.or.kr<br />
Molecular Monitoring of Microbes in Surface water <strong>and</strong><br />
Distribution System of Pipeline using PCR-DGGE<br />
Hee Suk LEE* 1 , Sun-Bok GEUN 2 <strong>and</strong> Min Jeoung KIM 1<br />
J-2<br />
1 2<br />
K-water. KOTTI.<br />
*Corresponding author: lovealga@kwater.or.kr<br />
The objective of this study is to monitor the microorganisms in various<br />
surface water <strong>and</strong> distribution system of pipeline in the use of Denaturation<br />
Gradient Gel Electrophoresis (DGGE) system. Five major surface water<br />
(GR, DH, BS, IS, CJ) <strong>and</strong> two samples of distribution system (DB, US)<br />
in the pipeline were used. Nucleic acids of these samples were amplified<br />
using PCR <strong>and</strong> confirmed by DGGE using 8% polyacryamide gel. The<br />
microbial communities of samples which were taken in November were<br />
more diversified than in August. The microbial community in the<br />
distribution system was also observed <strong>and</strong> compared with the data from<br />
surface water. The diversity of microbial community in the distribution<br />
system is less than that of surface wate. The PCR-DGGE methods can<br />
be used as a useful tool to compare the microbial community <strong>and</strong> expect<br />
the variation of microbial community.<br />
Keywords: Monitoring, PCR-DGGE, Surface water<br />
J-3<br />
Growth Kinetics of Algae under the Influence of Antibiotics<br />
using Toxy-PAM<br />
Man-Gi CHO*, Pina KNAUFF, Katharina KRÄHLING, Thomas AMBROSI,<br />
Vera KEIL, Khalida MANSOURI, Daniel ASCHENBRENNER,<br />
Binod PRASAD <strong>and</strong> Thiyam GENERAL<br />
Department of Bio-Chemical <strong>Engineering</strong>, Dongseo University Graduate<br />
School,Busan 617-716, Republic of Korea.<br />
*Corresponding author: mgcho@gdsu.dongseo.ac.kr<br />
With increasing importance of micro algae in the pharmacy, medicine,<br />
environmental engineering <strong>and</strong> further fields, it is necessary to investigate<br />
growth conditions <strong>and</strong> potential growth inhibitors. Genetical modifications<br />
of micro algae strains can be an alternative to meet the increasing dem<strong>and</strong><br />
of valuable compounds from micro algae. Thus the following work with<br />
respect to suspectibility of micro algae Tetraselmis suecica, Marine<br />
Chlorella sp. <strong>and</strong> Haematococcus pluvialis against antibiotic was carried<br />
out to investigated the growth kinetics under influence of antibiotics.<br />
Different concentration were applied <strong>and</strong> the strains were incubated for<br />
7 days. The cell number, the optical density (OD) <strong>and</strong> the fluorescence<br />
activity (F0, Fm, Y) using Toxy PAM were measured.The cell number<br />
<strong>and</strong> the optical density are continuously increasing with the time. The<br />
fluorescence activity appears independent of the antibiotic concentration.<br />
Keywords: Microalgae, Antibiotics <strong>and</strong> Fluorescence, Growth Kinetic
Peptide Self-assembled Simultaneous Formation of<br />
Organic-inorganic Hybrid Spheres <strong>and</strong> Their Potential<br />
Applications<br />
Jungok KIM 1 <strong>and</strong> Hor-Gil HUR* 1,2<br />
1<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>, Gwangju Institute of Science<br />
<strong>and</strong> Technology, Gwangju, Korea. 2 J-4<br />
International Environmental Research Center,<br />
Gwangju Institute of Science <strong>and</strong> Tehnology, Gwangju, Korea.<br />
*Corresponding author: kjungok@gist.ac.kr<br />
The hybrid structures composed of organics <strong>and</strong> inorganic nanoparticles<br />
has been attractive due to their improved properties <strong>and</strong> stabilities<br />
compared to homogeneous materials. Despite some advances, there are<br />
still limited works on the simultaneous assembly of biological molecules<br />
(e.g. peptide) <strong>and</strong> inorganic nanoparticles into well-designed<br />
supramolecular structures under aqueous conditions. In this study, hybrid<br />
spheres containing peptides <strong>and</strong> gold nanoparticles has been<br />
simultaneously synthesized in water using AG4 peptides which acted as<br />
a reducing agent to guide the nucleation <strong>and</strong> growth of gold nanoparticles<br />
<strong>and</strong> a precursor to form sphere-like structure by self-assembly where the<br />
size of hybrid spheres is precisely controlled by adjusting the operating<br />
temperature. The self-assembled peptide spheres remain stable even after<br />
selective removal of the gold nanoparticles by iodide etching. The surface<br />
of gold nanoparticles has been functionalized using thiol group linked<br />
to biomolecules. The ability to synthesize nanoparticle <strong>and</strong> self-assembled<br />
peptide structures with controlled size <strong>and</strong> composition in an<br />
environmental benign way will allow fabricating new class of<br />
multi-functional organic-inorganic hybrid superstructures for various<br />
biomedical <strong>and</strong> electronic applications.<br />
Keywords: peptide, self-assembly, gold nanoparticle<br />
Enhancement of Biomass <strong>and</strong> Fatty acids of Dunaliella salina<br />
According to the Culture Medium Compositions Containing the<br />
Animal Hormone, Steroids<br />
Mi-Kyung KIM* 1 , Yun-Sam KIM 2 , Dong-Mok LEE 3 <strong>and</strong> Inho CHOI 3<br />
1<br />
EcoPhycoTech, Marine Science Research Center, Yeungnam University,<br />
Gyeongsan 712-749, Korea. 2 Dept. of Biology, Yeungnam University, Gyeongsan<br />
712-749, Korea. 3 J-5<br />
School of Biotechnology, Yeungnam University, Gyeongsan<br />
712-749, Korea.<br />
*Corresponding author: mkkim@yu.ac.kr<br />
The main issues are focused on how to enhance efficiently the biomass<br />
for algal industrialization as a future bioresource: supplemental nutrients,<br />
pharmaceutical products, bioremediation, biofuel <strong>and</strong> reduction of<br />
greenhouse gas (CO2) emissions. The extracts of the wasted pig-testis<br />
(PTSE) contain steroid hormones - nondrolone (55%), testosterone (28%),<br />
<strong>and</strong>rostenedione (1%), estradiol (5%), estrone (4%), progesterone (7%)<br />
or the ones of pig-placenta (PPSE) containing nondrolone (13%),<br />
testosterone (0.2%), <strong>and</strong>rostenedione (1.7%), estradiol (2%), estrone<br />
(16.5%) <strong>and</strong> progesterone (66.6%), respectively. These extracts were<br />
added to the D media according to the different ratios of 5 ng/ml, 50<br />
ng/ml, 500 ng/ml for PTSE, 10 ng/ml, 100 ng/ml, 1 ug/ml for PPSE,<br />
respectively. The additions of PTSE of 5 ng/ml <strong>and</strong> PPSE of 10 ng/ml<br />
increased the cells densities of 250% <strong>and</strong> 200% , while the PTSE of<br />
500 ng/ml <strong>and</strong> PPSE of 1 ug/ml increased the fatty acids of 5500% <strong>and</strong><br />
8260% for Dunaliella salina, respectively. The amount of DHA(C22:6)<br />
of D. salina was two-fold more accumulated in PTSE of 500 ng/ml (22.3%)<br />
<strong>and</strong> PPSE of 1 ug/ml (24.4%) than control D medium. The mechanism<br />
of gene expression by animal steroids should be clarified in D. salina<br />
cultured in the different hormone compositions.<br />
Keywords: Steroid, Fatty acids, Dunaliella salina<br />
2011 International Symposium & Annual Meeting<br />
Effect of Biosurfactant-Producing Bacterium on Mineralization<br />
of Wastewater Contaminated with Petroleum by Petroleum-<br />
Degrading Bacterial Community<br />
Bo Young JEON 1 , Jun-Young YI 1 , Il Lae JUNG 2 <strong>and</strong> Doo Hyun PARK* 1<br />
1<br />
Department of Chemical <strong>and</strong> Biological <strong>Engineering</strong>, Seokyeong University,<br />
Seoul 136-704, Korea. 2 J-6<br />
Department of Radiation Biology, Environmental<br />
Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon<br />
305-353, Korea.<br />
*Corresponding author: baakdoo@skuniv.ac.kr<br />
Serratia sp. was isolated from riverbed soil in Jungrang-cheon <strong>and</strong><br />
cultivated with a mineral-based soybean oil medium. Serratia sp. grew<br />
in the mineral-based soybean oil medium but did not grow in a<br />
mineral-based kerosene-diesel medium. The petroleum-degrading bacteria<br />
used in this study--Acinetobacter sp., Pseudomonas sp., Paracoccus sp.,<br />
<strong>and</strong> Cupriavidus sp.--were isolated from a specially designed enrichment<br />
culture. The mineralization efficiency of wastewater contaminated with<br />
kerosene <strong>and</strong> diesel (WKD) by the PDBC was increased significantly by<br />
addition of the crude biosurfactant. The mineralization efficiency of the<br />
WKD was also enhanced by co-culture of Serratia sp. <strong>and</strong> PDBC. The<br />
temperature gradient gel electrophoresis (TGGE) pattern for the 16S rDNA<br />
variable region extracted from the co-cultured Serratia sp. <strong>and</strong> PDBC<br />
in the WKD at 2 nd day of incubation time was identical to that at the<br />
8 th day of incubation. In conclusion, the co-culture of Serratia sp. <strong>and</strong><br />
PDBC is an applicable technique for the mineralization of wastewater<br />
contaminated with petroleum, which may substitute for a biosurfactant.<br />
Acknowledgement This work was supported by the New &Renewable<br />
Energy of the KETEP grant funded by the Korea governmental Ministry<br />
of Knowledge Economy (2010T1001100334)<br />
Keywords: Biosurfactant, Serratia sp. petroleum-degrading bacteria, mixed<br />
culture, TGGE<br />
J-7<br />
Effect of Electrochemical Pulse on Microbial Community<br />
Variation <strong>and</strong> Growth of Crops<br />
Doo Hyun PARK, Jun-Young YI <strong>and</strong> Ji-Won CHOI*<br />
Department of Biological <strong>Engineering</strong>, Seokyeong University, Seoul 136-704,<br />
Korea.<br />
*Corresponding author: baakdoo@skuniv.ac.kr<br />
Electrochemical pulse generated from DC 2, 4, 6, 8, 10 V of electricity<br />
was induced to farm soil, in which lettuces were cultivated. The pulse<br />
intervals were controlled by exchange of electrode polarity at intervals<br />
of 10s. Distance between electrodes was adjusted to 10 cm, by which<br />
the unit of electric intensity charged to the farm soil was 200, 400, 600,<br />
800, <strong>and</strong> 1,000 mV per cm, respectively. Bacterial community diversity<br />
was analyzed <strong>and</strong> compared by temperature gradient gel electrophoresis<br />
<strong>and</strong> species identity of the bacterial community was determined based<br />
on the sequence homology of 16S-rDNA variable region that was extracted<br />
from TGGE gel. Lettuce growth was not activated in the electrochemical<br />
pulsed condition at 200 <strong>and</strong> 400 mV/cm of electrochemical potential but<br />
activated at 600, 800, <strong>and</strong> 1,000 mV of electrochemical potential in<br />
proportion to the electric intensity. Acknowledgement This research was<br />
supported by Seokyeong University in 2010<br />
Keywords: electrochemical pulse, bacterial community, soil bacteria<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
397
Characterization of Phosphate-Solubilizing Bacteria Isolated from<br />
Soil<br />
Ah Ra CHO 1 , Jae Gon KANG 2 , Hoon Seok KANG 2 <strong>and</strong> Dae Ook KANG* 1<br />
1<br />
Department of Biochemistry <strong>and</strong> Health Science, Changwon National University,<br />
Changwon 641-773, Korea. 2 J-8<br />
Korea Biochemical Inc., Igok-ri, Munsan-eup, Jinju<br />
660-841, Korea.<br />
*Corresponding author: biochem1@changwon.ac.kr<br />
Bacteria capable of forming halos on Pikovskaya's agar medium were<br />
isolated from soil as an effort to develop microbial agent for the amendment<br />
of inorganic phosphate-accumulated soil. Five bacterial isolates were<br />
finally selected <strong>and</strong> identified using biochemical <strong>and</strong> molecular taxonomy.<br />
Four isolates were identified as Acinetobacter calcoaceticus <strong>and</strong> the one<br />
Pantoea sp. Phosphate-solubization activity gradually increased for 7-day<br />
growth, which was verified by reciprocal decrease in elctronic conductance.<br />
The optimal temperature <strong>and</strong> initial pH were 30 o C <strong>and</strong> 5.0 - 6.0,<br />
respectively. The addition of 3% of NaCl had no effect on cell growth.<br />
We selected optimal C-, N-source <strong>and</strong> inorganic salts. The Pantoea sp.<br />
showed 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. We<br />
examined 2D protein expression profies of three isolates between normal<br />
<strong>and</strong> salt stressed (4% NaCl) culture condition.<br />
Keywords: ACC deaminase, electronic conductance, optimal temperature,<br />
pH, phosphate-solubilizing bacteria,<br />
Laboratory Evaluation of Entomopathogenic Fungi for Control<br />
of Green Peach Aphid Myzus persicae<br />
Van Hanh VU* 1 , Thi Thuy Duong LE 2 , Dinh Thi QUYEN 2 <strong>and</strong> Keun KIM 3<br />
1<br />
Institute of Biotechnology, Vietnam Academy of Science <strong>and</strong> Technology, Hanoi<br />
10600, Vietnam/ Department of Bioscience <strong>and</strong> Biotechnology, The University of<br />
Suwon, Hwaseong-si 445-743, Korea. 2 Institute of Biotechnology, Vietnam<br />
Academy of Science <strong>and</strong> Technology, Hanoi 10600, Vietnam. 3 J-9<br />
Department of<br />
Bioscience <strong>and</strong> Biotechnology, The University of Suwon, Hwaseong-si 445-743,<br />
Korea.<br />
*Corresponding author: vvhanh@ibt.ac.vn<br />
Twenty seven entomopathogenic fungal strains were isolated from<br />
various infected aphids. One fungus exhibited the highest virulence<br />
towards green peach aphids, Myzus persicae. The isolate conformed most<br />
closely to Lecanicillium sp. based on the sequence of internal transcribed<br />
spacer region of its 28S rDNA, <strong>and</strong> thus was designated as Lecanicillium<br />
sp. Le85. The laboratory bioassay for the aerial conidia of Le85 was<br />
performed for the control of M. persicae. After six days of application<br />
with 1×10 8 aerial conidia/ml, at 20~25 o C <strong>and</strong> relative humidity of 75~85%,<br />
the mortality of aphid by Le85 was about 97% with LT50 of 3.2 days.<br />
Meanwhile, at 26~31 o C <strong>and</strong> relative humidity of 60~70%, the evaluations<br />
of Le85 on potted cabbages showed an aphid mortality ranging from 82<br />
to 95%. Significant conidiation was observed on the aphid cadavers<br />
infected with Le85. Undoubtedly, the results suggest that Le85 can be<br />
used as a biocontrol agent for aphid M. persicae on cabbages in<br />
greenhouses.<br />
Keywords: Myzus persicae, entomopathogenic fungi, Lecanicillium sp.,<br />
398 www.kormb.or.kr<br />
Glucose Utilization <strong>and</strong> Electricity Generation Using Genetically<br />
Modified Shewanella Strains<br />
DongGeon CHOI 1 , Sae Bom LEE 1 , In Geol CHOI 2 <strong>and</strong> In Seop CHANG* 1<br />
1<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>, Gwangju Institute of Science<br />
<strong>and</strong> Technology(GIST), 261 Cheomdan gwagi-ro, Buk-gu, Gwangju 500-712,<br />
Korea. 2 J-10<br />
College of Life Science <strong>and</strong> Biotechnology, Korea University, Anam-dong,<br />
Sungbuk-gu, Seoul 136-701, Korea.<br />
*Corresponding author: ischang@gist.ac.kr<br />
Genomic sequence analyses of Shewanella strains have been studied<br />
for carbon metabolism as well as their physiological characteristics. Even<br />
though Shewanella strains seem to have glycolysis pathways <strong>and</strong> enzymes<br />
as central carbon metabolism, several key enzymes are missing.<br />
Shewanella strains also have only one set of phosphotransferase system<br />
to uptake simple sugar like glucose through the membrane. So, it is unclear<br />
that Shewanella strains can utilize six-carbon carbohydrates (e.g. glucose)<br />
as carbon <strong>and</strong> energy source. In this study, Shewanella oneidensis MR-1<br />
was metabolically engineered to have the ability of glucose uptake <strong>and</strong><br />
utilization as carbon <strong>and</strong> energy source. Glucokinase (glk) <strong>and</strong><br />
glucose-facilitated diffusion protein (glf) of Zymomonas mobilis were<br />
cloned into a broad host range plasmid <strong>and</strong> introduced to S. oneidensis<br />
MR-1 to help glucose uptake <strong>and</strong> utilization. This recombinant Shewanella<br />
strain was used for single cell based MFC system in order to confirm<br />
the glucose utilization <strong>and</strong> concomitant with current production. The<br />
results indicated that cloned genes (glf <strong>and</strong> glk) were functional for glucose<br />
utilization in S. oneidensis MR-1 wild type. We also found that this<br />
modification was effective to two Shewanella strains, S. putrefaciens<br />
CN-32 <strong>and</strong> S. loihica PV-4.<br />
Keywords: Shewanella, Glucose, Electricity<br />
Investigation of C<strong>and</strong>idate Strains for Application of Microbial<br />
Electrosynthesis<br />
Saebom LEE 1 , Dong Geon CHOI 1 , In Geol CHOI 2 <strong>and</strong> In Seop CHANG* 1<br />
1<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>, Gwangju Institute of Science<br />
<strong>and</strong> Technology (GIST), 261 Cheomdan-gwagiro, Buk-gu, Gwangju 500-712,<br />
Korea. 2 J-11<br />
College of Life Sciences <strong>and</strong> Biotechnology, Korea University,<br />
Anam-dong, Sungbuk-ku, Seoul 136-701, Korea.<br />
*Corresponding author: ischang@gist.ac.kr<br />
Reducing power which is obtained from microbial metabolism is used<br />
to produce several valuable chemicals such as ethanol, butanol, <strong>and</strong> acetate.<br />
Microbial electrosynthesis is a system that produces chemicals using<br />
electron. In microbial electrosynthesis, electrons supplied from electrode<br />
are accumulated to inner cell <strong>and</strong> are used to produce the reducing power.<br />
The accumulated reducing power can be used to produce chemicals.<br />
Geobacter <strong>and</strong> Shewanella strains, which are well known as metal reducing<br />
bacteria, can deliver electrons to outer metal ions through the arrangement<br />
of cytochrome C protein in the cell membrane. Recently, as reverse<br />
electron flow of Shewanella strain was revealed, it was known to move<br />
outer electrons to inside through cytochrome C protein. Based on this<br />
point, we selected c<strong>and</strong>idate strains which have cytochrome C protein<br />
in the cell membrane <strong>and</strong> can produce valuable chemicals though microbial<br />
reaction. We finally selected Carboxydothermus hydrogenoformans strain<br />
Z-2901 <strong>and</strong> Dethiobacter alkaliphilus AHT 1 which have cytochrome C<br />
protein in extracellular <strong>and</strong> are able to produce valuable chemicals. Two<br />
c<strong>and</strong>idate strains will be used for microbial electrosynthesis.<br />
Keywords: Microbial electrosynthesis; Reverse electron flow
Production of Bio-gas From Moon Jellyfish (Aurelia aurita)<br />
Ji-Youn KIM 2 <strong>and</strong> Jae-Hwa LEE* 1<br />
J-12<br />
1<br />
College of Medical <strong>and</strong> Life Science, Silla University, Busan 617-736, Korea.<br />
2<br />
Medical <strong>and</strong> Life Science, Silla University, Busan 617-736, Korea.<br />
*Corresponding author: kjy6504@hanmail.net<br />
The recent study of a jellyfish has caused a hindrance to sea fishery.<br />
Presently, jellyfish is thrown away through a separator system in the sea.<br />
The objective of this work was to produce bio-gas from Aurelia aurita<br />
using anaerobic digestion. The bio-gas includes the hydrogen or the<br />
methane gas. It relates that jellyfish is effectually changed into the<br />
renewable energy. In previous study, optimal culture conditions of jellyfish<br />
have been established, <strong>and</strong> based on these, <strong>and</strong> a research including the<br />
jellyfish amount of addition, the method of treatment, the pH control.<br />
It optimized with the jellyfish amount of addition with the amount of<br />
3 g. And the effect of substrate heat treatment, sonication treatment <strong>and</strong><br />
freeze treatment conditions on hydrogen <strong>and</strong> methane gases production<br />
in batch experiments. The optimal treatment condition was found to be<br />
Freeze treated at -70℃ for 20 min. Laboratory-scale experiments were<br />
carried out in completely mixed bioreactors, 5 L capacity with 4 L worked.<br />
Optimal amount of cultivation for organics in the step of pretreatment<br />
was 120 mL Aurelia aurita <strong>and</strong> used 40 mL wastewater sludge.<br />
Keywords: Bio-gas, Jellyfish (Aurelia aurita), Anaerobic digestion<br />
Ag Coated Hydroxyapatite [+Polyacrylonitrile(nanofiber),<br />
+Polyurethane(film)] Inhibit Growth of Microorganisms<br />
Kwang-Hwan JHEE* 1 , Jung-Hyun KIM 1 , Byung-Gil MIN 2 <strong>and</strong><br />
Ju-Yeon KIM 2<br />
1<br />
Dept. of Applied Chemistry, Kumoh National Institute of Technology, Gumi<br />
730-701, Korea. 2 J-13<br />
Dept. of Nano-Bio Textile <strong>Engineering</strong>, Kumoh National<br />
Institute of Technology, Gumi 730-701, Korea.<br />
*Corresponding author: khjhee@kumoh.ac.kr<br />
Hydroxyapatite (HAp) is a calcium phosphate ceramic that is used as<br />
a biomaterial. It is chemically similar to the mineral component of bones<br />
<strong>and</strong> hard tissues in mammals <strong>and</strong> its ability to form a strong bond to<br />
the human hard tissue. Silver (Ag) <strong>and</strong> Ag-containing compounds have<br />
been exhibited strong antimicrobial activity. Ag-based antimicrobials draw<br />
much attention because of the low toxicity of the active Ag ion to human<br />
cells. We studied the antibacterial effects of Ag containing nanofibers<br />
(HAp/Polyacrylonitrile, PAN) <strong>and</strong> films (HAp/Polyurethane, PU). PAN,<br />
PU <strong>and</strong> HAp were used for the preparation of electrospinning nanofibers<br />
<strong>and</strong> films. Ag containing nanofibers <strong>and</strong> films were prepared by<br />
electrospinned with Ag [2.5-5% in DMF(fiber) or H2O/MEK(film)]<br />
together or soaked into Ag containing DMF(fiber) or H2O/MEK(film)<br />
solution (1,000 ppm). The antimicrobial effects were investigated by paper<br />
disc or shaking flask method against E. coli, K. pneumoniae (Gramnegative)<br />
<strong>and</strong> S. aureus (Gram-positive). Our data exhibited high<br />
antibacterial activity against gram-negative <strong>and</strong> gram-positive both.<br />
Therefore Hap coated Ag containing fibers <strong>and</strong> films might be a good<br />
c<strong>and</strong>idate for the medicinal textile like b<strong>and</strong>age <strong>and</strong> baby clothes using<br />
their antimicrobial activity.<br />
Keywords: Hydroxyapatite, Antimicrobial activity, Silver<br />
2011 International Symposium & Annual Meeting<br />
Oriental Medicines Showing Antifungal Activity Against<br />
Raffaelea quercus-mongolicae<br />
Yongsub YI* 1,2 <strong>and</strong> Jaejoung KIM 2<br />
J-14<br />
1<br />
Department of Herbal Cosmetics Science, Hoseo University, Asan, Korea.<br />
2<br />
Department of Biochemistry, Hoseo University, Asan, Korea.<br />
*Corresponding author: yongsub@hoseo.edu<br />
Oriental medicines showing antifungal activity against tree wilt disease<br />
pathogen, Raffaelea quercus-mongolicae, were studied. 16 oriental<br />
medicines were extracted <strong>and</strong> assayed antifungal activity against the<br />
pathogen by disk diffusion assay. Coptidis Rhizoma showed high<br />
antifungal activity against Raffaelea quercus-mongolicae. Extract of<br />
Coptidis Rhizoma was fractioned by LC chromatography. Each fractions<br />
was assayed antifungal activity by disk diffusion assay. More research<br />
was studied by HPLC <strong>and</strong> MASS analysis.<br />
Keywords: antifungal activity, Raffaelea quercus-mongolicae<br />
Eco-Monitoring of Algal Blooming in Sooyoung Bay<br />
Hui-Chih WANG 2 , Cora HANESCH 1 , Annalena PEDREIRO 1 ,<br />
Nadine ROYLA 1 , Jessica SCHULZE 1 , Binod PRASAD 1 , Byung-Gook LEE 3<br />
<strong>and</strong> Man-Gi CHO* 1<br />
1<br />
Department of Bio-Chemical <strong>Engineering</strong>, Dongseo University Graduate School,<br />
Busan 617-716, Republic of Korea. 2 Department of Bioprocess <strong>Engineering</strong>,<br />
Friedrich-Alex<strong>and</strong>er-University of Erlangen Nürnberg, Busan Campus 618-230,<br />
Republic of Korea. 3 J-15<br />
Institue of Ambient Intelligence, Dongseo University<br />
Graduate School, Busan 617-716, Republic of Korea.<br />
*Corresponding author: mgcho@gdsu.dongseo.ac.kr<br />
The purpose of this work is to diagnose the algal blooming based on<br />
Global Positioning System (GPS) <strong>and</strong> the fluorescence measurement using<br />
ToxY-Pam in the Sooyoung Bay. Especially there the samples were taken<br />
because of the observed algal blooming. The used device measured the<br />
minimum fluorescence (F0) <strong>and</strong> the maximum quantum yield of<br />
photosystem II (Fv/Fm) from the taken samples.The samples were taken<br />
in December <strong>and</strong> March. The Chlorophyll Fluorescence values of the<br />
samples collected in December were lower as compared to March.<br />
Therefore the collected data can be used for forecasting the onset of algal<br />
blooming.<br />
Keywords: Algal Blooming, Fluorescence, Eco-Monitoring<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
399
Thermodynamic <strong>and</strong> Kinetic Analyses of the Binding of a Bacterial<br />
Expansin (BsEXLX1) to Plant Cell Wall Components<br />
Injung KIM 2 , Hyeok-Jin KO 2 , In-Geol CHOI 2 , Douglas S. CLARK 3 ,<br />
Tae-Wan KIM 3 <strong>and</strong> Kyoung Heon KIM* 1<br />
1<br />
School of Life Sciences <strong>and</strong> Biotechnology, Korea University, Seoul 136-713,<br />
Korea. 2 Biotechnology, Korea University, Seoul 136-713, Korea. 3 J-16<br />
Energy<br />
Biosciences Institute, Department of Chemical &Biomolecular <strong>Engineering</strong>,<br />
University of California, Berkeley, CA, 94720, USA.<br />
*Corresponding author: khekim@korea.ac.kr<br />
BsEXLX1 is a protein belonging to expansin family, found in a soil<br />
bacterium, Bacillus subtilis, <strong>and</strong> was previously shown to possess the<br />
synergistic activity with cellulase on the cellulose hydrolysis. In this study,<br />
binding characteristics of BsEXLX1 to plant cell wall components were<br />
thermodynamically <strong>and</strong> kinetically investigated. At 4°C, the values of<br />
binding constants, such as Ka <strong>and</strong> [No], obtained from Langmuir isotherm<br />
were 6.73´10 6 M -1 <strong>and</strong> 54 nmol/g cellulose, respectively. Also, the binding<br />
of BsEXLX1 to cellulose was found to be an endothermic but spontaneous<br />
reaction (∆Gº=-36.2 kJmol -1 <strong>and</strong> ∆Hº=57 kJmol -1 ). Generally, the<br />
BsEXLX1 exhibited an enhanced binding efficiency under acidic<br />
conditions. Also, the amount of bound protein increased in the presence<br />
of NaCl. Against cellopentaose <strong>and</strong> cellohexaose, the binding activity of<br />
BsEXLX1 was negligible in the isothermal titration calorimetry. Against<br />
the pretreated lignocellulosic biomass with different compositions of<br />
cellulose, hemicelluloses <strong>and</strong> lignin, the preferential binding of BsEXLX1<br />
to lignin was observed. This finding could be applicable in cellulose-based<br />
pulp <strong>and</strong> textile industries or biofuel production for using the bacterial<br />
expansin as a non-blocking agent to enhance the accessibility of cellulose<br />
to enzymes.<br />
Keywords: Bacterial expansin, Cellulose, Lignocellulose<br />
Screening of an Esterase/lipase-Producing Bacteria from<br />
Gastropods<br />
Jung-Hee WOO* 1 , Sun Mee HONG 1 , Jun-Tae KIM 1 , Tae-Hyung KWON 1 ,<br />
Hyun-Soo RHO 2 , Jong-Shik KIM 1 , Nyun-Ho PARK 1 <strong>and</strong><br />
Choong-Gon KIM 1<br />
1<br />
1Gyeongbuk Institute for Marine Bio-Industry (GIMB), Uljin 767-813,<br />
Gyeongbuk, Korea. 2 J-17<br />
2East Sea Research Institute, Korea Ocean Research<br />
&Development Institute (KORDI), Uljin 767-813, Gyeongbuk, Korea.<br />
*Corresponding author: jhwoo@gimb.or.kr<br />
Esterases <strong>and</strong> lipases are enzymes that catalyze the hydrolysis of<br />
carboxyl esters <strong>and</strong> are found in various organisms including animals,<br />
plants, fungi <strong>and</strong> bacteria. These enzymes have increased the<br />
biotechnological interest for a number of industrially significant<br />
biotransformations, either as hydrolytic or as synthetic catalysts.<br />
Previously we reported that bacteria isolated 127 strains from six<br />
gastropods F. oregonensis, N. cumingi, N. constricta, C. fulmen, V.<br />
emphaticus, JG in June to December 2010 were studied to find out the<br />
bacterial assemblages associated with local gastropod collected in Uljin,<br />
the East Sea, Korea. Esterase/lipase-producing bacteria were isolated from<br />
gastropods. To screen for lipolytic activity, various agar medium<br />
supplemented with 1% tributyrin (TBN) <strong>and</strong> tricaprylin (TCN) were used.<br />
We found that 56 isolates showed lipolytic activity on TBN agar plate<br />
<strong>and</strong> 22 isolates showed lipolytic activity on TCN agar plate<br />
(esterase-producing strains= 34, lipase-producing strains= 22). This study<br />
presents a first example which discovered esterases/lipases from<br />
gastropods successfully. [This work was supported by the Marine <strong>and</strong><br />
Extreme Genome Research Center Program, Ministry of L<strong>and</strong>, Transport<br />
<strong>and</strong> Maritime Affairs, Republic of Korea.]<br />
400 www.kormb.or.kr<br />
J-18<br />
Survival of Escherichia coli O157:H7 <strong>and</strong> Listeria in Manure<br />
Compost in Korea<br />
Sook Young JANG, Sunggi HEU, Eunjung ROH, Dong Hwan LEE,<br />
Yang Kyun KIM, Mukesh KUMAR, Shweta MALHOTRA, Jong Chul YUN<br />
<strong>and</strong> Kyu Seok JUNG*<br />
Microbial Safety Division, National Academy of Agricultural Science, RDA,<br />
Suwon, 441-707, Korea.<br />
*Corresponding author: win258@korea.kr<br />
On-farm contaminations through contaminated manure were considered<br />
likely sources of the pathogen for several outbreak. Of particular concern<br />
are bacteria causing human food-borne illness such as Salmonella species<br />
<strong>and</strong> E. coli O157:H7. The objective of this study was to quantify the<br />
level of environmental impact of temperature on E. coli O157:H7 <strong>and</strong><br />
Listeria survival in manure compost. Commercial manure compost<br />
(manure 60%, sawdust 40%) was inoculated with E. coli O157:H7 <strong>and</strong><br />
Listeria. Composts were incubated at four different temperature (10, 25,<br />
35, <strong>and</strong> 55℃) <strong>and</strong> consistent moisture content (55%). Samples had been<br />
collected during 49 days depending on the given conditions. E. coli<br />
O157:H7 <strong>and</strong> Listeria persisted for up to 1 day in compost at 55℃ <strong>and</strong><br />
49 days at three different temperature (10, 25 <strong>and</strong> 35℃). It is noted that<br />
as the temperature increased, the number of E. coli O157:H7 <strong>and</strong> Listeria<br />
gradually decreased over time. The results indicate that E. coli O157:H7<br />
<strong>and</strong> Listeria persisted longer under low temperature condition. This study<br />
will provide useful <strong>and</strong> practical guidelines to applicators of soil in<br />
deciding appropriate h<strong>and</strong>ling <strong>and</strong> time frames for l<strong>and</strong> application for<br />
sustainable agriculture.<br />
Keywords: E. coli O157:H7, Listeria, survival<br />
Microbiological Safety of Gamma-Irradiated Porcine Skin for<br />
the Development of Dressing Material<br />
Jong-Il CHOI 1 , Eu-Ri JO 1 , Pil-Mun JUNG 1 , Jae-Hun KIM 1 ,<br />
Beom-Suk SONG 1 , Jae-Kyung KIM 1 , Jong-Heum PARK 1 , Tae-Woon KIM 2<br />
<strong>and</strong> Ju-Woon LEE* 1<br />
J-19<br />
1<br />
Team for Radiation Food Science <strong>and</strong> Biotechnology,Advanced Radiation<br />
Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185.<br />
2<br />
Rion-Biotechnology Co.,wanju-gun 565-902, Korea.<br />
*Corresponding author: sjwlee@kaeri.re.kr<br />
In this study, sensitivities of bacteria <strong>and</strong> virus in porcine skin by gamma<br />
irradiation were evaluated to develope the optimum sterilization condition<br />
for the development of dressing material. Escherichiacoli (E.coli) <strong>and</strong><br />
Bacillus substilis (B. substilis) were used as model pathogens <strong>and</strong><br />
inoculated 106-107log CFU/g. As model virus from porcine, porcine<br />
parvovirus (PPV), bovine viral diarrhoea virus (BVDV) <strong>and</strong> poliovirus<br />
were inoculated 105-106TCID50/g into porcine skin. The D10 value of<br />
E.coli was determined as 0.25±0.1 kGy. B. substilis showed lower<br />
radiation sensitivity as D10 was 3.88±0.3 kGy. The D10 values of PPV,<br />
BVDV <strong>and</strong> poliovirus were determined as 1.73±0.2, 3.81±0.2 <strong>and</strong> 6.88±0.3<br />
kGy, respectively. These results suggest that gamma irradiation can reduce<br />
the risk of contamination <strong>and</strong> offer the basic information to inactivate<br />
the pathogens as microbiologically safe dressing material of porcine skin.<br />
Keywords: radiation sensitivity, bacteria <strong>and</strong> virus, porcine skin
Optimal Culture Conditions of Microalgae Haematococcus<br />
pluvialis<br />
Hyun-Jin PARK 1 , Young-Hwa KIM 2 <strong>and</strong> Jae-Hwa LEE* 1<br />
1<br />
Department of Bioscience <strong>and</strong> Biotechnology, College of Medical <strong>and</strong> Life<br />
Science, Silla University, Busan 617-736, Korea. 2 J-20<br />
Department of Pharmaceutical<br />
<strong>Engineering</strong>, College of Medical <strong>and</strong> Life Science, Silla University, Busan<br />
617-736, Korea.<br />
*Corresponding author: phj0386@naver.com<br />
The green microalgae Haematococcus pluvialis (H. pluvialis) produces<br />
large amounts of astaxanthin. The morphological change of H. pluvialis<br />
from green vegetative cells to red resting cyst cells is accompanied by<br />
an enhancement in astaxanthin production. To find out high mass culture<br />
conditions of H. pluvialis, some important growth factors, such as<br />
temperature, initial pH, culture media, CO2, <strong>and</strong> air effect were tested.<br />
The optimal growth condition was found to be as follows: temperature<br />
35℃, initial pH 6.5, 4X MBBM medium. Maximum growth rate of H.<br />
pluvialis was obtained at a 5% CO2(0.81 g/L), air(1.06 g/L) environment.<br />
From these conditions, it could be possible to cultivate H. pluvialis for<br />
large-scale high density.<br />
Keywords: optimal culture, Haematococcus pluvialis, marine microalga<br />
Heterologous Expression of Metagenome-Derived Wax Ester<br />
Synthase in Escherichia coli <strong>and</strong> Arabidopsis thaliana.<br />
Nam-hee KIM 1 , Ji Hye PARK 1 , Eun Sook CHUNG 2 , Jin Cheol KIM 3 ,<br />
Myung Hwan LEE 1 , Weixin TAO 2 , Eul Chul HWANG 1 , Jai Heon LEE 2 <strong>and</strong><br />
Seon Woo LEE* 1,2<br />
1<br />
Department of Applied Biology, Dong- A University, Busan 604-714, Republic<br />
of Korea. 2 Department of Medical Bioscience, Dong- A University, Busan<br />
604-714, Republic of Korea. 3 J-21<br />
Chemical Biotechnology Research Center, KRICT,<br />
Deajeon 305-600, Republic of Korea.<br />
*Corresponding author: seonlee@dau.ac.kr<br />
We have previously selected a metagenome clone carrying an ORF<br />
encoding a wax ester synthase (WES) like protein. Deduced amino acid<br />
sequences of the ORF exhibited 30% <strong>and</strong> 22% identities to WES from<br />
Acinetobacter baylyi <strong>and</strong> Arabidopsis thaliana, respectively. The<br />
WES-like protein was functional as a wax ester synthase, when wes gene<br />
was expressed in Escherichia coli fed by fatty alcohol, 10 mM of<br />
hexadecanol, as a precursor for wax ester synthesis. The unique wax esters<br />
produced in E. coli carrying wes gene were identified as wax esters of<br />
various chain lengths by GC-MS analysis. The wes gene was also<br />
introduced into a model plant, Arabidopsis thaliana, to produce wax esters<br />
in a plant system. Transgenic plants carrying the wes gene were obtained<br />
by Agrobacterium-mediated transformation. Expression of wes gene in<br />
the transgenic plants was confirmed by RT-PCR. Production of wax esters<br />
<strong>and</strong> other phenotypic changes in the transgenic plants are under<br />
investigation.<br />
2011 International Symposium & Annual Meeting<br />
J-22<br />
Community Structure Analysis of Cyanobacteria using 16S rRNA<br />
<strong>and</strong> Phycocyanin Gene in Korean Reservoirs<br />
Kyoung-Hee OH, Seung-Hee SHIN, Eun-Hye PARK, Mi-Na YU <strong>and</strong><br />
Young-Cheol CHO*<br />
Dept. of Environmental <strong>Engineering</strong>, Chungbuk National University, Cheongju<br />
361-763, Korea.<br />
*Corresponding author: choy@cbnu.ac.kr<br />
The community structure of cyanobacteria was examined with 16S<br />
rRNA <strong>and</strong> phycocyanin intergenic spacer (PC-IGS) genes as target.<br />
Although DGGE analysis with 16S rRNA gene reflected the changes of<br />
cyanobacterial community structure with fluctuations of environmental<br />
factors, the resolution was restricted to the genus level because of its<br />
homogeneity. On the other h<strong>and</strong>, analysis of PC-IGS gene sequences<br />
showed that this gene can classify cyanobacteria in strain level, indicating<br />
it is considered as an adequate target gene to analyze the community<br />
structure of cyanobacteria in the environmental samples. Moreover, the<br />
PC-IGS gene sequences of Microcystis strains in the investigated reservoirs<br />
were divided into two clades <strong>and</strong> each clade exclusively included toxicor<br />
non-toxic Microcystis, proposing PC-IGS gene can be used to monitor<br />
toxigenic Microcystis at the cyanobacterial bloom events. [Supported by<br />
grants from National Science Foundation of Korea]<br />
Keywords: Cyanobacteria, PC-IGS, Community structure<br />
J-23<br />
Ethanol Fermentation using Main Sugar in Laminaria japonica<br />
Geun Hyub KIM, Sung-Mok LEE <strong>and</strong> Jae-Hwa LEE*<br />
Department of Bioscience <strong>and</strong> Biotechnology, College of Medical <strong>and</strong> Life<br />
Science, Silla University, Busan 617-736, Korea.<br />
*Corresponding author: jhalee@silla.ac.kr<br />
The excessive use of fossil fuels has recently been causing many<br />
problems around the world such as global warming. Therefore, alternative<br />
energy source has been of interest, much research as been performed in<br />
order to develop alternative energy. This study examined the production<br />
of bio-ethanol from the brown-algae main sugar compound. The result<br />
of ethanol fermentation used four-bacteria, ethanol production of<br />
Zymomonas mobilis KCTC 1534 with different carbon sources (alginate,<br />
laminaran, mannitol <strong>and</strong> Laminaria japonica) confirmed 2.31 g/L, 1.32<br />
g/L <strong>and</strong> 2.36 g/L respectively. The maximum cell growth with laminaran<br />
reached 1.42 (O.D. 600 nm), but other substrate was not significant<br />
increases in the fermentation period. The maximum cell growth of<br />
Zoogloea ramigera KCCM 40423 was reached approximately 2.61 <strong>and</strong><br />
2.37 (O.D. 600 nm) with mannitol <strong>and</strong> laminaran, respectively. And then<br />
ethanol production was 0.97 <strong>and</strong> 0.236 g/L. Therefore, Laminaran like<br />
to be limitations to what ethanol production, but can consume for cell<br />
growth increase. Streptomyces venezuelae KCCM 40310, Paracoccous<br />
denitrificans KCCM 40267 was not detected ethanol production, but cell<br />
growth was significant increases.<br />
Keywords: ethanol fermantation, Laminaria japonica, polysaccharides<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
401
Identification of Novel Sugar Pathway Genes <strong>and</strong> their<br />
Organization from an Insect Gut Metagenome<br />
Chang-Muk LEE* 1 , Sang-Hong YOON 1 , Soo-Jin KIM 2 , So-Hyeon SEO 1 ,<br />
Young-Seok LEE 1 , Jung-Keun PARK 1 , Han-Chul KANG 1 <strong>and</strong><br />
Bon-Sung KOO 1<br />
J-24<br />
1<br />
Functional Biomaterial Division, National Academy of Agricultural Science,<br />
2<br />
Rural Development Administration, Suwon 441-707, Korea. National<br />
Agrobiodiversity Center, Academy of Agricultural Science, Rural Development<br />
Administration, Suwon 441-707, Korea.<br />
*Corresponding author: changmuk@rda.go.kr<br />
Insect gut symbiotic microbiota play essential roles in the recycling<br />
various organic materials, <strong>and</strong> may thus be exploited as a simple system<br />
for processing daily food wastes. To detect the contributing microbial<br />
genes, we have constructed 92,544 metagenomic fosmid clones using the<br />
larval intestinal microbiome from black soldier fly, Hermetia illucens.<br />
The hydrolytic enzymes encoded by uncultured microorganisms were<br />
subjected to substrate hydrolysis analysis. Initial screening of the libraries<br />
revealed a clone that uses carboxymethyl cellulose as a sole carbon source.<br />
Functional screening <strong>and</strong> metagenome walking with shot-gun libraries<br />
revealed two novel gene clusters conferring multiple sugar pathways. The<br />
cluster was 36kb in size <strong>and</strong> transcribed as polycistronic units. They<br />
include glycosidase-related sugar pathway genes that are arranged as sugar<br />
transferase genes, glycan processing genes, transporter genes, <strong>and</strong> putative<br />
transcription factors. The presence of insertion sequences between clusters<br />
suggests that the metagenome-derived clone has acquired the specific<br />
organization by lateral gene transfer [Supported by grants from the Rural<br />
Development Administration project number PJ0067032011].<br />
Keywords: Metagenome, Hermetia illucens gut, Glycosidase<br />
Development of Transgenic Daphnia magna Expressing Enhanced<br />
green Fluorescent Protein<br />
Thai Hoang LE 1 , Sung-Kyu LEE 2 , Yang-Hoon KIM 3 <strong>and</strong> Jiho MIN* 1<br />
1<br />
Department of Bioprocess <strong>Engineering</strong>, Chonbuk National University, 664-14<br />
Duckjin-dong, Jeonju 561-756, South Korea. 2 Toxicology Research Center, Korea<br />
Research Institute of Chemical Technology, Daejon, 305-600, Korea. 3 J-25<br />
School of<br />
Life Science, Chungbuk National University 410 Sungbong-Ro, Heungduk-Gu,<br />
Cheongju 361-763, South Korea.<br />
*Corresponding author: jihomin@jbnu.ac.kr<br />
To establish a novel system for a rapid detection of environmental stress,<br />
a transgenic Daphnia magna able to express a green fluorescent protein<br />
has been created. A 32bp DNA fragment containing the 18s-ribosome<br />
constitute promoter of daphnia was generated by in-vitro annealing the<br />
commercial primers at 80 o C. This target DNA was inserted ahead to the<br />
gfp gene in the EGFP vector (Enhanced Green Flourescent Protein). The<br />
successful recombinant plasmid confirmed by the DNA sequencing was<br />
introduced into the earlier developing-state daphnia eggs (round shape)<br />
using microinjection technique. After 48h incubation in the 20 o C chamber,<br />
the injected eggs able to develop to juveniles were observed at the 488nm<br />
wavelength in the confocal fluorescent microscope to confirm the presence<br />
of green fluorescent protein. Our results show that the green fluorescent<br />
protein was substantially found in the upper part of the transgenic<br />
organisms (e.g., head, back) while no fluorescent signal was detected in<br />
the control organism. It demonstrated that a transgenic Daphnia magna<br />
expressing green fluorescent protein was firstly generated <strong>and</strong> would be<br />
promising to build a novel testing system for easily monitoring<br />
environmental stress.<br />
Keywords: Daphnia magna, transgenic, fluorescent<br />
402 www.kormb.or.kr<br />
Isolation <strong>and</strong> Characteristics of Antagonic Microorganisms as<br />
Nematicide Against Root-knot Nematode of Fruit Vegetables<br />
(Oriental Melon <strong>and</strong> Cucumber)<br />
Beam Soo KIM 1 , Keun-Hyung LEE 1 , Young-Won KANG 1 , Chang-Su KIM 1 ,<br />
Se-Ho MYEONG 1 , Joung-Ja KWON 2 , Chung-Sig CHOI 3 <strong>and</strong><br />
Gi-Seok KWON* 1<br />
1<br />
School of Bioresource Sciences, Andong National University, Andong 760-749,<br />
Korea. 2 Cheongsong Agricultural Technology Center Cheong Song Gun,<br />
CheongSong 763-802, Korea. 3 J-26<br />
Bio Industry Institute, HansBio Co.,Ltd. Andong<br />
760-380, Korea.<br />
*Corresponding author: suyaa18@hanmail.net<br />
This study was conducted to screen the antagonic microorganisms on<br />
root-knot nematode <strong>and</strong> to utilize in its control. The effect of nematocidal<br />
activity of antagonic microorganisms were tested on the nematicide of<br />
root-knot nematode, Showing nematocidal activities of 88 strains isolated<br />
from various fruit Vegetables field <strong>and</strong> soil. In vitro study, all tested<br />
treatments had nematicidal effect on nematode juveniles 24 <strong>and</strong> 48 hours<br />
from exposures in the 24-well plate. Among them, BS-17, BS-51(2),<br />
BS-51(3), BS-52(2), <strong>and</strong> BS-84(2) were isolated. The highest percentage<br />
of nematode mortality was achieved by application of BS-17(96.38%),<br />
BS-51(2)(94.11%), BS-51(3)(95.38%), BS-52(2)(94.59%), BS-84(2)<br />
(98.48%). Under greenhouse conditions, isolated antagonic microorganisms<br />
treatments were effective in reducing population numbers of root-knot<br />
nematode <strong>and</strong> root gall formation in compared to non-treatments.<br />
Keywords: Root-knot nematode, Meloidogyne. Antagonic microorganisms,<br />
Nematocidal activity, Ne<br />
Effect of Plant Growth-Promoting Rhizobacteria on Growth of<br />
Chinese cabbage Plant under Salt Stress<br />
Hee Sun KOOK 1 , Gun-Woon LEE 1 , Song-Joong YUN 2 , Jong-Chan CHAE 1<br />
<strong>and</strong> Kui-Jae LEE* 1<br />
J-27<br />
1<br />
Division of Biotechnology, Chonbuk national University, Iksan 570-752, korea.<br />
2<br />
Division of Agriculture Life sciences, Chonbuk National University, Chonju<br />
561-765, Korea.<br />
*Corresponding author: k722954@naver.com<br />
To gain underst<strong>and</strong>ing of responses to salinity stress by rhizobacteria<br />
at physiological level, we isolated 15 strains of plant growth-promoting<br />
bacteria from soil samples taken in the reclaimed tidel<strong>and</strong>s. These 15<br />
strains have been identified as Pseudomonas, Herbaspirillum, Enterobacter,<br />
Serratia, <strong>and</strong> Stenotrophomonas by 16S rRNA gene sequence analysis.<br />
With the exception of Serratia sp., all strains can product indole acetic<br />
acid (IAA) <strong>and</strong> IBA (butyric acid). Among the selected 15 strains, eight<br />
strains only showed 1-aminocyclopropane-1-carboxylate (ACC)<br />
deaminase activity <strong>and</strong> rhizobacterial ACC deaminases were detected by<br />
polyclonal anti-ACCD antibody. To evaluate whether rhizobacteria<br />
inhabiting saline environments confer the resistance against salt stress to<br />
Chinese cabbage plant, cabbage seeds were sown in soil inoculated with<br />
respective bacterial strains. And then, the early seedling growth was<br />
examined under 0 or 100 mM NaCl solution treatment for 2 weeks after<br />
germination. Results of our study exhibited that inoculation with selected<br />
strains increased the shoot length ranging from 107 to 141%, compared<br />
to that of the uninoculated control. These results indicate that the<br />
rhizobacterial stains had the potential to promote the growth of cabbage<br />
plant under saline condition.<br />
Keywords: plant growyh-promoting rhizobacteria (PGPR), Chinese cabbage,<br />
NaCl, Salt stress
J-28<br />
Characterization of Microbial Community of the Ammonia<br />
Oxidizing Enrichment Culture from Wastewater Treatment Plants<br />
Hyoun Duk JUNG <strong>and</strong> Eun Young LEE*<br />
Department of Environmental Engineerin <strong>and</strong> Environmental <strong>Microbiology</strong>,<br />
Suwon University, Gyeonggi 445-743, Korea.<br />
*Corresponding author: ley@suwon.ac.kr<br />
Ammonia oxidizing enrichment culture carried from S wastewater<br />
treatment plant was performed in the AOB medium. The microbial<br />
community in wastewater treatment plants were characterized using<br />
potential carbon availability method. For the Ecoplate (Biolog, USA)<br />
analysis, those samples were inoculated <strong>and</strong> cultured at 20℃. Color<br />
changes of individual well in the Ecoplate according to cell growth were<br />
measured at 595nm <strong>and</strong> average well color development (AWCD) was<br />
calculated. The order of AWCD values of S1(enrichment culture),<br />
S2(aeration tank), S3(effluent)<strong>and</strong> S4(influent) were 0.36,1.18,1.12 <strong>and</strong><br />
0.98, respectively.The Shannon indies were 2.31(S1), 3.35 (S2),3.37(S3)<br />
<strong>and</strong> 3.38(S4),respectively. In addition, Bacterial community was<br />
characterized using 16S rDNA PCR <strong>and</strong> denaturing gradient gel<br />
electrophoresis (DGGE) fingerprinting method. the similarities of DGGE<br />
fingerprints were 7% between S1 <strong>and</strong> S2, 11% between S1 <strong>and</strong> S3, 7%<br />
between S1 <strong>and</strong> S4, 11% between S2 <strong>and</strong> S3, 8% between S2 <strong>and</strong> S4,<br />
13% between S3 <strong>and</strong> S4 of the ammonia oxidizing enrichment culture<br />
from wastewater treatment plants.<br />
Keywords: DGGE, Ecoplate, wastewater<br />
Reduction of Tellurite <strong>and</strong> Formation of Tellurium Nanostructures<br />
by Shewanella oneidensis MR-1<br />
Dong-Hun KIM 1 <strong>and</strong> Hor-Gil HUR* 1,2<br />
J-29<br />
1 2<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>. International Environmental<br />
Analysis <strong>and</strong> Educational Center, Gwangju Institute of Science <strong>and</strong> Technology,<br />
Gwangju 500-712, Republic of Korea.<br />
*Corresponding author: donghun@gist.ac.kr<br />
The metalloid oxyanion tellurite(TeO3 2- ) is highly toxic to most<br />
organisms <strong>and</strong> specific mechanisms explaining its toxicity are not well<br />
known to date. In this study we determine the reduction of tellurite <strong>and</strong><br />
formation of tellurium by S. oneidensis MR-1. Strain MR-1 can reduce<br />
tellurite [TeO3 2- ] to elemental tellurium [Te(0)] under anaerobic conditions.<br />
This reduction result in the formation of unique crystalline Te<br />
nanoarchitectures as end products. The localization of needle-like shaped<br />
Te nanostructures are controlled by culture conditions. EDX <strong>and</strong> SAED<br />
indicate that nanostructures are composed entirely of Te <strong>and</strong> are crystalline.<br />
Furthermore, c-type cytochrome deletion mutants <strong>and</strong> r<strong>and</strong>om mutants<br />
from strain MR-1 shown lower reduction rate of tellurite than wild type<br />
<strong>and</strong> different morphologies on tellurite contained solid medium. Additional<br />
works needs for describe more details about molecular mechanisms of<br />
the tellurite reduction <strong>and</strong> formation of nanostructures. This research offers<br />
the potential to combine remediation of contaminants with the development<br />
of environmental friendly manufacturing pathways for novel biogenic<br />
nanominerals <strong>and</strong> recovery of rare metals from industrial waste water.<br />
Keywords: Keywords: Shewanella, Tellurium, dissimilatory metal-reducing<br />
bacteria (DMRB), nano structure<br />
2011 International Symposium & Annual Meeting<br />
J-30<br />
Improved Performance of a Microbial Electrolysis cell using a<br />
Membrane Electrode Assembly Cathode<br />
Yuhong JIA, Jae Hun RYU, Cho Hui KIM, Woo Kyung LEE,<br />
Thi Van Trinh TRAN, Hyo Lee LEE <strong>and</strong> Dae Hee AHN*<br />
Department of Environmental <strong>Engineering</strong> <strong>and</strong> Biotechnology, Myongji<br />
University, Yongin 449-728, Korea.<br />
*Corresponding author: dhahn@mju.ac.kr<br />
Microbial electrolysis cell is a device which can produce hydrogen gas<br />
from biomass through microbial catalyzed process <strong>and</strong> thus reduce the<br />
organic matter. For the real application in wastewater treatment, the<br />
scale-up of microbial electrolysis cell is an important issue but few tests<br />
were conducted with relatively large size. In this study, a 3.7 L microbial<br />
electrolysis cell equipped with a membrane electrode assembly cathode<br />
was designed <strong>and</strong> tested. The internal resistance was examined, hydrogen<br />
generation <strong>and</strong> organic removal performance was investigated under<br />
different conditions. An internal resistance of 7 Ω was examined from<br />
the reactor with a relatively large working volume. The MEC achieved<br />
a coulombic <strong>and</strong> cathodic efficiency of 55 % <strong>and</strong> 74 % respectively at<br />
an applied voltage of 1.2 V, results a maximum overall hydrogen efficiency<br />
of 41 %. This corresponding to a volumetric hydrogen production rate<br />
of 0.12 L-H2/LA-day (A = Anodic liquid volume) at a hydrogen yield<br />
of 1.64 mol H2/mol acetate <strong>and</strong> a current density of 2.8 A/m 2 . The<br />
hydrogen performance at voltages below 0.8 V was limited, which might<br />
be due to the high potential losses in large system. The results obtained<br />
in this study could help to further develop pilot-MEC for practical<br />
applications.<br />
Keywords: Microbial electrolysis cell, Hydrogen production, Membrane<br />
electrode assembly<br />
Anti-bacteria <strong>and</strong> Anti-fungal Effects of Herbal Oil made from<br />
Halvateria acuminata.<br />
Soo Ji KIM* 1 , L.M Udaya CABRAL 2 , Jin Young HONG 1 , Chang Wook<br />
JO 1 , Mi Hwa JUNG 1 , Young Hee KIM 1 <strong>and</strong> Jung Eun CHOI 1<br />
1<br />
Conservation Science Division, National Research Institute of Hultural Heritage,<br />
82 Munji-ro, Yuseong-gu, Daejeon, 305-380, Korea. 2 J-31<br />
Conservation <strong>and</strong><br />
Preservation Division, National Library <strong>and</strong> Documentation Services Board,<br />
No.14, Independence Avenue, Colombo-7, Sri Lanka.<br />
*Corresponding author: feeelsj@naver.com<br />
An Ola leaf manuscript, which consists of words carved on an Ola<br />
leaf <strong>and</strong> filled up with mixture of herbal oil made from Halvateria<br />
acuminata <strong>and</strong> charcoal, is one of the native writing media in Sri Lanka.<br />
According to historical records, Ola leaf manuscripts had been used from<br />
1 st century A.D to 18 th century A.D. From recent findings that Ola leaf<br />
manuscripts have been preserved well for the past 400years <strong>and</strong> are in<br />
good state of preservation, it is supposed that this oil preserves Ola leaves<br />
against environment factors <strong>and</strong> biological factors such as fungi <strong>and</strong><br />
insects. To evaluate the anti-biological susceptibility of the herbal oil,<br />
the molds isolated from wooden printing blocks in Janggyeong Panjeon<br />
of Haeinsa temple in Korea <strong>and</strong> bacteria isolated from Ola leaf are cultured.<br />
After spreading of microorganisms suspension on agar plate, a disk paper<br />
containing certain volume of herbal oil is put on the agar plate. From<br />
the experiment, it is found that the herbal oil exhibits a clear zone, which<br />
is optically clear <strong>and</strong> inhibits growth of microorganisms, against some<br />
molds <strong>and</strong> bacteria. The result indicates that herbal oil from the plant<br />
Halvateria acuminata has the anti-bacteria <strong>and</strong> anti-fungal properties.<br />
Keywords: herbal oil, anti-bacteria, anti-fungal<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
403
J-32<br />
Enhancement of Biohydrogen Production in [NiFe]-hydrogenase<br />
Expressing Recombinant Escherichia coli by Introducing Light<br />
Acceptor<br />
Younghwa JO, Jaoon Y.H. KIM, Byung Hoon JO <strong>and</strong> Hyung Joon CHA*<br />
Pohang University of Science <strong>and</strong> Technology, Pohang 790-784, Korea.<br />
*Corresponding author: hjcha@postech.ac.kr<br />
Biohydrogen production can be one of solutions for energy crisis.<br />
Previously, we successfully demonstrated the production of biohydrogen<br />
by constructing [NiFe]-hydrogenase-expressing recombinant E. coli.<br />
Although [NiFe]-hydrogenase showed relatively high oxygen-tolerance<br />
compare to other hydrogenases, still biohydrogen production efficiency<br />
was low. In this research, we introduced proteorhodopsin, light acceptor<br />
for prokaryote working as proton pump, into [NiFe]-hydrogenaseexpressing<br />
recombinant E. coli for utilizing light energy for producing<br />
biohydrogen. Proteorhodopsin need a chemical named retinal for working<br />
as proton pump by using light energy but E. coli does not have a pathway<br />
to synthesize it. In order to give E. coli an ability to synthesize retinal,<br />
we expressed five foreign genes expressing enzymes for retinal synthesis<br />
pathway. By introducing light accepting system into E. coli expressing<br />
[NiFe]-hydrogenase, we could get enhanced biohydrogen productivity.<br />
Keywords: Biohydrogen, [NiFe]-hydrogenase, proteorhodopsin<br />
Novel Probiotic Strains for American Foulbrood <strong>and</strong> Nosemosis<br />
of Honeybee<br />
Yoon Jeong JANG 1 , Byoung Joo SEO 1 , Irfan RATHER 1 , Uk Han CHOI 1 ,<br />
Jeonghuei LIM 1 , Kook Hee KANG 2 , Byoung Su YOON 3 <strong>and</strong><br />
Yong-Ha PARK* 1<br />
J-33<br />
1<br />
Department of Applied <strong>Microbiology</strong> <strong>and</strong> Biotechnology, Yeungnam University.<br />
2 3<br />
Korea Institute of Science <strong>and</strong> Technology Information, Daejeon. Department<br />
of Life Science, Kyonggi University, Suwon, Kyonggi-Do.<br />
*Corresponding author: joos02@ynu.ac.kr<br />
American Foulbrood of honeybees (Apis mellifera), caused by the<br />
Gram-positive bacterium Paenibacillus larvae is one of the most serious<br />
diseases affecting the larval <strong>and</strong> pupal stages of honeybees. Presently,<br />
Nosema ceranae <strong>and</strong> Nosema apis seems to be the main microsporidian<br />
infection in honeybees. The aim of this study was to evaluate lactic acid<br />
bacteria strains isolated from Kimchi <strong>and</strong> to study its inhibitory effects<br />
against Paenibacillus larvae subsp. larvae KCTC 3744 <strong>and</strong> Nosema<br />
trichoplusiae ATCC 30702. In all, 3,500 strains were screened <strong>and</strong> three<br />
highly active bacteria were selected <strong>and</strong> were identified as L. plantarum<br />
YML001, L. plantarum YML004, Leu. citreum KM20. The optical density<br />
of P. larvae KCTC 3744 at OD600nm showed drastic decrease in cell<br />
growth as compared to control. Three groups of honey bees (N=25) were<br />
given Nosema Spores at a concentration of 10,000 spores per ml with<br />
addition of 1% of supernatant of three strains. The bees were incubated<br />
at 37°C <strong>and</strong> samples were taken seven days post infection, <strong>and</strong> the amount<br />
of spores in the mid gut was counted. We found that the three strains<br />
inhibited the growth of P. larvae KCTC 3744 <strong>and</strong> N. trichoplusiae ATCC<br />
30702 <strong>and</strong> provided new perspective for probiotics of honeybees.<br />
Keywords: American Foulbrood, Nosemosis, Probiotics<br />
404 www.kormb.or.kr<br />
Analysis of Bacterial Community in Electrochemical Bioreactors<br />
Cultivated with Carbon Dioxide as Sole Carbon Source<br />
Doo Hyun PARK 2 , Jinsu KIM 2 , Bo Young JEON 2 <strong>and</strong> Il Lae JUNG* 1<br />
1<br />
Department of Radiation Biology, Environmental Radiation Research Group,<br />
Korea Atomic Energy Research Institute, Daejeon 305-353, Korea. 2 J-34<br />
Department<br />
of Biological <strong>Engineering</strong>, Seokyeong University, Seoul 136-704, Korea.<br />
*Corresponding author: baakdoo@skuniv.ac.kr<br />
Bacterial community was isolated from different sources that were forest<br />
soil, aerobic wastewater treatment reactant, anaerobic wastewater digestive<br />
reactant, farm field soils, mixture of isolated bacterial strains. Bacterial<br />
communities grown in the different bioreactors were analyzed by<br />
temperature gradient gel electrophoresis technique. Different bacterial<br />
communities were enriched <strong>and</strong> selected during cultivation with the<br />
electrochemical reducing power as a sole energy source. TGGE pattern<br />
is not an indicator to show all of the bacterial communities grown in<br />
the electrochemical bioreactors; however, the bacterial species capable of<br />
growing with carbon dioxide or surviving in the condition saturated with<br />
carbon dioxide may be enriched selectively. According to the bacterial<br />
species identified based on the sequence homology, various bacterial<br />
species may fix metabolically carbon dioxide. Acknowledgement This<br />
work was supported by the New &Renewable Energy of the Korea Institute<br />
of Energy Technology Evaluation <strong>and</strong> Planning (KETEP) grant funded<br />
by the Korea governmental Ministry of Knowledge Economy<br />
(2010T1001100334)<br />
Keywords: electrochemical reducing power, carbon dioxide fixation,<br />
Bacterial community diversity<br />
Screening of Protease-Producing Bacteria from Gastropods<br />
Jung-Hee WOO* 1 , Sun Mee HONG 1 , Jun-Tae KIM 1 , Jin-Wook LEE 1 ,<br />
Hyun-Soo RHO 2 , Jong-Shik KIM 1 , Nyun-Ho PARK 1 <strong>and</strong> Choong-Gon KIM 1<br />
1<br />
1Gyeongbuk Institute for Marine Bio-Industry (GIMB), Uljin 767-813,<br />
Gyeongbuk, Korea. 2 J-35<br />
2East Sea Research Institute, Korea Ocean Research<br />
&Development Institute (KORDI), Uljin 767-813, Gyeongbuk, Korea.<br />
*Corresponding author: jhwoo@gimb.or.kr<br />
Microbial proteases are the most important group of industrial enzymes<br />
such as food, detergent, pharmaceutical, <strong>and</strong> waste management industries.<br />
Previously we reported that bacteria isolated 127 strains from six<br />
gastropods F. oregonensis, N. cumingi, N. constricta, C. fulmen, V.<br />
emphaticus, JG in June to December 2010 were studied to find out the<br />
bacterial assemblages associated with local gastropod collected in Uljin,<br />
the East Sea, Korea. Protease-producing bacteria were isolated from<br />
gastropods. To screen for proteolytic activity, various agar medium<br />
supplemented with 0.5% skim milk were used. We found that 38 isolate<br />
strains showed proteolytic activity on skim milk agar plate (10 o C = 16<br />
strains, 25 o C = 22 strains). This study presents a first example which<br />
discovered microbial proteases from gastropods successfully. [This work<br />
was supported by the Marine <strong>and</strong> Extreme Genome Research Center<br />
Program, Ministry of L<strong>and</strong>, Transport <strong>and</strong> Maritime Affairs, Republic of<br />
Korea]<br />
Keywords: Protease, Proteolytic activity, Gastropods
J-36<br />
Simultaneous Quantification of Cyanobacteria <strong>and</strong> Microcystis<br />
spp. using Quantitative Realtime PCR<br />
Kyoung-Hee OH, Seung-Hee SHIN, Mi-Na YU, Eun-Hye PARK <strong>and</strong><br />
Young-Cheol CHO*<br />
Dept. of Environmental <strong>Engineering</strong>, Chungbuk National University, Cheongju<br />
361-763, Korea.<br />
*Corresponding author: choy@cbnu.ac.kr<br />
Microcystins are produced mainly by Microcystis sp. at large reservoirs<br />
in Korea. Therefore, quantification of Microcystis is required to monitor<br />
the potential for algal-toxin production. It is difficult to count Microcystis,<br />
however, because these mostly form scum. In this study, new method<br />
was developed to quantify cyanobacteria <strong>and</strong> Microcystis simultaneously<br />
using quantitative realtime PCR. The primers <strong>and</strong> probe, cyanobacteriaspecific<br />
<strong>and</strong> Microcystis-specific primers <strong>and</strong> Microcystis-specific TaqMan<br />
probe, were designed from the conserved regions of 16S ribosomal RNA<br />
gene sequences for cyanobacteria <strong>and</strong> Microcystis, respectively. It was<br />
proved as an adequate <strong>and</strong> reproducible method to quantify cyanobacteria<br />
<strong>and</strong> Microcystis in the environmental samples. It is expected that the use<br />
of this method enhances the accuracy of Microcystis counts <strong>and</strong> reduces<br />
the time involved in the analysis of samples by microscopy. Also, this<br />
method enables us to monitor the formation of cyanobacterial mass<br />
occurrences in situ <strong>and</strong> to reveal the factors promoting the growth of<br />
Microcystis. [Supported by grants from National Science Foundation of<br />
Korea]<br />
Keywords: Microcystis, Real-time PCR, Simultaneous quantification<br />
J-37<br />
GFP Tagging of p-Cresol Methylhydroxylase in Pseudomonas<br />
alkylphenolia <strong>and</strong> Its Dominant Expression in Colonies<br />
CHO AH RA <strong>and</strong> Kyoung LEE*<br />
Dept. of <strong>Microbiology</strong>, Changwon National University, Kyongnam 641-773,<br />
Korea.<br />
*Corresponding author: 0506joara@hanmail.net<br />
In this study, the chromosome-encoded pcuRCAXB genes that are<br />
required for p-cresol degradation have been identified by using the newly<br />
constructed Green fluorescence protein (GFP)-based promoter probe<br />
transposon in the long-chain alkylphenol degrader, Pseudomonas<br />
alkylphenolia. The deduced amino acid sequences of the genes showed<br />
the highest identities at the levels of 65-93% compared to those in the<br />
databases. The transposon was identified to be inserted in the pcuA gene<br />
with the promoterless gfp gene being under the control of the pcu catabolic<br />
gene promoter. The expression of GFP was positively induced by p-cresol<br />
<strong>and</strong> was about 10 times higher by cells grown on agar than those in<br />
liquid culture. This result indicates that P. alkylphenolia additionally<br />
possesses a protocatechuate ortho cleavage route for p-cresol degradation<br />
that is dominantly expressed in colonies.<br />
Keywords: Pseudomonas, transposon<br />
2011 International Symposium & Annual Meeting<br />
Isolation <strong>and</strong> Characteristics of Entomopathogenic Fungi<br />
Beauveria bassiana as Insectide Agent Against Greenhouse<br />
Ahitefly(Trialeurodes vaporariorum)<br />
Chang-Su KIM 2 , Keun-Hyung LEE 3 , Young-Ho NAM 3 ,<br />
Gyeong-Muk KANG 3 , Jin-Won KIM 4 <strong>and</strong> Gi-Seok KWON* 1<br />
1<br />
Departmentl of bioresource Sciences Andong Nation University, Andong<br />
760-749, Korea. 2 안동대학교. 3 School of bioresource Sciences, Andong Nation<br />
University, Andong 760-749, Korea. 4 J-38<br />
Yecheon Agricuture Technology Center,<br />
Yecheon 757-802, Korea.<br />
*Corresponding author: ckdtn0629@hanmail.net<br />
The object of this study is to use Entomopathogenic fungi B.bassiana<br />
as agent Greenhouse whitefly(Trialeaarodes vaporariorum). Each 4 strains<br />
were prepared as follows : B.bassiana was parceled by Rural Development<br />
Administration(J157&J216), Gyougbuk Agricutural Reserch&Extension<br />
Service(S186) Yecheon Agricultuer Technology Center(B.K). Chitinase,<br />
protease, lipase are important <strong>and</strong> involved in the host invasion. In vitro,<br />
plate assay was used to observe the each enzyme. And chitinase was<br />
observed by three strains. Also, Protease, lipase was observed by all strains.<br />
And activity measurement of enzyme is in progress. In vivo, one control<br />
plot <strong>and</strong> each four treatment plots was treated to 50 Greenhouse whiteflys<br />
in a capsicum. After a few days, B.bassiana spore(60g/1L) that was<br />
mixtuerd the yolk of an egg spraied due volta 500ml in each treatment<br />
plots <strong>and</strong> spraied a Neoseiulus californicus in the control plot. Later,<br />
indicated Greenhouse whitefly that enticed yellow flypaper trap <strong>and</strong><br />
compared control plot with treatment plots. Consequently, B.bassiana was<br />
a predominant <strong>and</strong> confirmed as insecticide agent against Greenhouse<br />
whitefly.[ This study supported by Technology of Development program<br />
for Agriculture <strong>and</strong> Foresty, Gyeongbuk 2011]<br />
Keywords: Beauveria bassiana, Entomopathogenic, Greenhouse whitefly<br />
J-39<br />
Effect of Nitrate on the Electricity Generation of Microbial Fuel<br />
cell<br />
Jae Kyung JANG*, Jung Eun CHOI, Young Sun RYOU, Sung Hyoun LEE,<br />
Jong Goo KIM, Youn Koo KANG, Young Hwa KIM <strong>and</strong> Hyung Mo LEE<br />
National Academy of Agricutural Science.<br />
*Corresponding author: jkjang1052@korea.kr<br />
The microbial fuel cells (MFCs) can convert to useful electricity from<br />
artificial wastewater including nitrogen source such as nitrate (NO3 - ) <strong>and</strong><br />
also to treat this wastewater. Electrons <strong>and</strong> protons produced by metabolic<br />
process of microorganisms in the anode compartment transfer to cathode<br />
through the out circuit <strong>and</strong> membrane, respectively. However, when the<br />
nitrate is included in the wastewater (fuel), it can act the electron donor.<br />
It will decrease the current value. These studies were conducted to<br />
investigate the influence of nitrate concentration on the current generation.<br />
Sodium acetate was used as electron donor <strong>and</strong> the chemical oxygen<br />
dem<strong>and</strong> (CODcr) of the wastewater was around 250 mg L -1 . The microbial<br />
fuel cells produced a current about 9 mA stably when 249.2±4.6 mg/L<br />
artificial wastewater as COD including only acetate without nitrate was<br />
fed. When nitrate from 31.4±0.4 to 123.3±0.1 mg/L was fed into MFCs<br />
with sodium acetate, the current generation decreased from 9.97±0.06 to<br />
0.20±0.01 mA according to increase the nitrate concentration. However<br />
nitrate concentration was removed up to 96% as added nitrate of 123.3±0.1<br />
mg/L. These results showed that nitrate could behave the electron acceptor<br />
in the anode compartment.<br />
Keywords: Microbial fuel cell, nitrate, electricity generation<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
405
Bioelectrochemical Fixation of CO2<br />
Bo Young JEON 1 , Ji-Won CHOI 1 , Il Lae JUNG 2 <strong>and</strong> Doo Hyun PARK* 1<br />
1<br />
Department of Chemical <strong>and</strong> Biological <strong>Engineering</strong>, Seokyeong University,<br />
Seoul 136-704, Korea. 2 J-40<br />
Department of Radiation Biology, Environmental<br />
Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon<br />
305-353, Korea.<br />
*Corresponding author: baakdoo@skuniv.ac.kr<br />
For the isolation of CO2-fixing bacteria using electrochemical reducing<br />
power (ERP), a plate-type electrochemical bioreactor (PEB) was employed.<br />
Soluble neutral red (NR) was used as an electron mediator for<br />
carbonate-basal agar medium (CBAM) prepared in the PEB. For test of<br />
bacterial CO2 fixation using the ERP, a single-compartmented<br />
electrochemical bioreactor (SCEB) was employed. NR immobilized in the<br />
graphite felt cathode (NR-cathode) was used as an electron mediator for<br />
carbonate-basal broth medium (CBBM) prepared in the SCEB. Two<br />
bacterial genera capable of electrochemically fixing CO2 were isolated<br />
using the PEB <strong>and</strong> cultivated using the SCEB. The isolated bacterial species<br />
were 99% identified with Achromobacter sp. <strong>and</strong> Alcaligenes sp.<br />
Approximate 150 ml of CO2 per day was consumed by 300 ml of the<br />
mixed culture of Achromobacter sp. <strong>and</strong> Alcaligenes sp. grown<br />
electrochemically in the SCEB. Acknowledgement This work was<br />
supported by the New &Renewable Energy of the Korea Institute of Energy<br />
Technology Evaluation <strong>and</strong> Planning (KETEP) grant funded by the Korea<br />
governmental Ministry of Knowledge Economy (2010T1001100334)<br />
Keywords: CO2-fixing bacteria, neutral red, electrochemical reducing<br />
power, Alcaligenes sp., Achromobacter sp.<br />
Inactivation of Chloramphenicol <strong>and</strong> Florfenicol by a Novel<br />
Chloramphenicol Hydrolase from Alluvial Soil Metagenome<br />
Weixin TAO 1 , Myung Hwan LEE 2 , Jing WU 2 , Nam Hee KIM 2 ,<br />
Jin-Cheol KIM 3 , Eul Chul HWANG 2 <strong>and</strong> Seon-Woo LEE* 1,2<br />
1<br />
Department of Medical Bioscience, Dong-A University, Busan 604-714, South<br />
Korea. 2 Department of Applied Biology, Dong-A University, Busan 604-714,<br />
South Korea. 3 J-41<br />
Chemical Biotechnology Research Center, KRICT, Daejeon<br />
305-343, South Korea.<br />
*Corresponding author: Seonlee@dau.ac.kr<br />
In a previous study, EstDL136 isolated from alluvial soil metagenome<br />
was redefined as chloramphenicol (Cml) acetate esterase, on account of<br />
its capability of Cml reactivation that against Cml acetyltransferase (CAT).<br />
Further investigation of Cml catalytic activity in the absence of CAT<br />
revealed that Cml was metabolized by EstDL136. The metabolite was<br />
identified as p-nitrophenylserinol by LC-MS <strong>and</strong> 1 H-NMR, suggesting a<br />
promiscuous amidase activity of the enzyme. Purified EstDL136 catalyzed<br />
Cml hydrolysis when they were co-incubated. Residues were detected by<br />
HPLC, which showed that Cml was gradually hydrolyzed <strong>and</strong> the<br />
hydrolysate, p-nitrophenylserinol was concurrently generated. EstDL136<br />
also hydrolyzed florfenicol (Ffl), the synthetic florinated analog of Cml<br />
that was resistant to CAT. Therefore, Escherichia coli was tolerant to<br />
Cml or Ffl when estDL136 was expressed, <strong>and</strong> duration of the lag phase<br />
was highly correlated with the concentrations of supplemented antibiotics<br />
(Cml, Ffl), indicating that estDL136 mediated the amphenicols resistance.<br />
This study primarily reported a novel bacterial Cml hydrolase that also<br />
hydrolyzes Ffl, conferring Cml <strong>and</strong> Ffl resistance to the E. coli.<br />
Keywords: Chloramphenicol, Florfenicol, Hydrolase<br />
406 www.kormb.or.kr<br />
J-42<br />
Efficacy of Combined Application of Entomopathogenic Fungal<br />
Conidia <strong>and</strong> Botanical Insecticides for Aphid Control<br />
Tuan anh PHAM <strong>and</strong> Keun KIM*<br />
Dept. of Bioscience <strong>and</strong> Biotechnology, Univ. of Suwon, Gyeonggi-do 445-743,<br />
Korea.<br />
*Corresponding author: ptanh140380@yahoo.com<br />
Some botanical insecticides prepared from neem, derris, garlic <strong>and</strong><br />
chinaberry were evaluated for the influence of the botanical insecticides<br />
on susceptibility of conidia of Beauveria bassiana by the germination<br />
ratio of conidia. The results demonstrated that botanical insecticides<br />
significantly reduced the germination ratio of conidia <strong>and</strong> it could not<br />
be mixed with the conidia in oil-based formulation for long-term storage.<br />
After that, the virulence of oil-based formulation of conidia in combination<br />
with botanical insecticides against aphid Myzus persicae was examined<br />
with detached leaf test. The botanical insecticide was mixed with the<br />
oil-formulated conidia just before spraying <strong>and</strong> the result showed that<br />
the combined application of two insecticides significantly increased the<br />
mortality of the aphid.<br />
Keywords: Beauveria bassiana conidia, botanical insecticides, aphid control<br />
Seasonal Escherichia coli Genotypes Determined by Horizontal<br />
Fluorophore-Enhanced rep-PCR (HFERP) in the Yeongsan River<br />
Basin of South Korea<br />
Jeonghwan JANG 1 , Tatsuya UNNO 1 , Seung Won LEE 1 , Kyung Hwa CHO 1 ,<br />
Michael J. SADOWSKY 3,4 , GwangPyo KO 5 , Joon Ha KIM 1 <strong>and</strong><br />
Hor-Gil HUR* 1,2<br />
1<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>, Gwangju Institute of Science<br />
<strong>and</strong> Technology, Gwangju, Republic of Korea. 2 International Environmental<br />
Research Center, Gwangju Institute of Science <strong>and</strong> Technology, Gwangju, Republic<br />
of Korea. 3 Department of Soil, Water <strong>and</strong> Climate, University of Minnesota, St.<br />
Paul, Minnesota, USA. 4 BioTechnology Institute, University of Minnesota, St. Paul,<br />
Minnesota, USA. 5 J-43<br />
Department of Environmental Public Health, Seoul National<br />
University, Republic of Korea.<br />
*Corresponding author: hghur@gist.ac.kr<br />
Seasonal <strong>and</strong> spatial variation in the genotypic diversity of 3,480 Escherichia<br />
coli isolates obtained from the Yeongsan River basin in South Korea was investigated<br />
by using the horizontal fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting<br />
technique. The MDS analysis, done based on HFERP DNA fingerprints, showed<br />
that E. coli isolates obtained in October through December clustered tightly, while<br />
those obtained in other sampling periods were more genetically diverse. SOMs<br />
analysis, done using the 10 most frequently-isolated E. coli genotypes, showed the<br />
occurrence of season-specific E. coli genotypes <strong>and</strong> the main SOMs clusters were<br />
most influenced by temperature, strain diversity, <strong>and</strong> biochemical oxygen dem<strong>and</strong>.<br />
Diversity among E. coli genotypes tended to decrease as water temperature decreased,<br />
<strong>and</strong> the numbers of E. coli genotypes observed in urban area were greater, more<br />
diverse, <strong>and</strong> less dependent on water temperature than those obtained from agricultural<br />
areas. This work was supported by the National Research Foundation of Korea Grant<br />
funded by the Korean Government (MEST) (NRF-R1A5A004-2010-0029224), <strong>and</strong><br />
also by the Korea Environmental Technology &Industry Institute (KEITI) as the<br />
Next Generation Eco Innovation Technology Development Project.<br />
Keywords: Escherichia coli, Yeongsan River, Horizontal<br />
Fluorophore-Enhanced rep-PCR
Antifungal Property of Microorganisms against Korea Oak Wilt<br />
Pathogen, Raffaelea quercus-mongolicae<br />
Yongsub YI* 1,2 <strong>and</strong> Jaejoung KIM 2<br />
J-44<br />
1<br />
Department of Herbal Cosmetics Science, Hoseo University, Asan, Korea.<br />
2<br />
Department of Biochemistry, Hoseo University, Asan, Korea.<br />
*Corresponding author: yongsub@hoseo.edu<br />
This study was performed to investigate the antifungal materials to<br />
inhibit growth of Raffaelea quercus-mongolicae. Microbes were tested to<br />
screen antifungal activity. We found microbes showing antifungal activity.<br />
Two hundreds strains were isolated <strong>and</strong> incubated on the same fresh media.<br />
Five strains, SG 1-9, 1-12, SG 2-8, 2-10, <strong>and</strong> 2-17, were collected by<br />
determination of antifungal activity. Out of the five strains, organic<br />
compounds <strong>and</strong> proteins of SG2-17 strain broth was extracted for<br />
determination of antifungal activity. The organic compound showed<br />
antifungal activity, <strong>and</strong> the proteins extract didn’t show it. The 16S rDNA<br />
sequences of the above five strains were determined by the sequencing<br />
analysis. The 16S rDNA sequences were analyzed by the homology of<br />
BLAST program at NCBI. The homology search showed that SG 1-9<br />
<strong>and</strong> 1-12 had 98% homology with Streptomyces cinnamoneus <strong>and</strong> 98%<br />
homology with Burkholderia cepacia, SG 2-8, 2-10, <strong>and</strong> 2-17 had 99%<br />
homology with Streptomyces fradiae, 97% homology with Staphylococcus<br />
epidermidis, <strong>and</strong> 99% homology with Staphylococcus epidermidis.<br />
Keywords: antifungal activity, Raffaelea quercus-mongolicae<br />
J-45<br />
Optimization of the Conditions to make Solid Medium for<br />
Cyanobacteria<br />
Seung-Hee SHIN, Mi-Na YU, Eun-Hye PARK, Kyoung-Hee OH <strong>and</strong><br />
Young-Cheol CHO*<br />
Dept. of Environmental <strong>Engineering</strong>, Chungbuk National University, Cheongju<br />
361-763, Korea.<br />
*Corresponding author: choy@cbnu.ac.kr<br />
The conditions to make solid medium for cyanobacteria, especially<br />
focused on the kind <strong>and</strong> concentration of the solidifying agents, was<br />
optimized to cultivate these organisms on solid medium, which is<br />
accounted as the easiest way to establish the axenic culture. Two different<br />
culture media, modified CB <strong>and</strong> BG-11, supplemented with bactoagar,<br />
edible-agar, or agarose, were used. The bactoagar <strong>and</strong> edible-agar were<br />
washed with water to remove the potential inhibitors. The concentrations<br />
of the agents were 0.4% or 1.0%. In order to check the efficiency of<br />
the culture conditions, the number <strong>and</strong> diversity of cyanobacteria on the<br />
solid media were examined. The cyanobacterial numbers on the media<br />
with rinsed agars were not significantly different from those on the<br />
unprocessed ones, indicating the inhibitors in these solidifying agents<br />
cannot be removed with the simple washing. Although the numbers of<br />
colonies on the BG-11 media were generally higher than those on the<br />
modified CB media, lots of heterotrophic bacterial colonies were found.<br />
Based on the number <strong>and</strong> diversity of cyanobacteria on the media, we<br />
concluded that the modified CB medium with 0.4% of agarose is the<br />
adequate condition for solid culture of these organisms. [Supported by<br />
grants from Eco-Technopia 21 Project]<br />
Keywords: Cyanobacteria, Solid culture, Agarose<br />
2011 International Symposium & Annual Meeting<br />
J-46<br />
Qualitative Detection of Model Bacteria Expressing Iturin <strong>and</strong><br />
Mop Cyclase <strong>and</strong> Microbial Community Analyses in Soils During<br />
the Cultivation of Plants<br />
Sung Eun KIM, Won-Sik CHOI, Jae Sun MOON <strong>and</strong> Sung Uk KIM*<br />
Korea Research Institute of Bioscience <strong>and</strong> Biotechnology, Daejeon 305-806,<br />
Korea.<br />
*Corresponding author: kimsu@kribb.re.kr<br />
To detect the microorganisms released into the environment at the<br />
molecular level, a recombinant strain P. fluorescens MX1 <strong>and</strong> B. subtilis,<br />
which have mop cyclase <strong>and</strong> iturin biosynthetic genes in the chromosome,<br />
respectively, were employed as model bacteria. Using specific fusion<br />
primers <strong>and</strong> the TaqMan probes, quantitative detections of model bacteria<br />
by real-time PCR were performed using genomic DNAs extracted from<br />
the soils of cucumber or tomato plants cultivated in pots <strong>and</strong> the<br />
greenhouse at intervals of three weeks for a six month period. The genomic<br />
DNAs from P. fluorescens MX1 <strong>and</strong> B. subtilis, except for probe DNAs,<br />
did not show any signals on the Southern blotting, indicating that the<br />
horizontal gene transfer from model bacteria to soil bacteria did not occur<br />
during the period monitored in pots <strong>and</strong> the greenhouse. In addition, the<br />
T-RFLP patterns of genomic DNAs obtained from the soils of both pots<br />
<strong>and</strong> the greenhouse suggest that the changes of microbial communities<br />
seem to be associated with the seasonal change in the soils. Thus, our<br />
data suggest that the bacteria used as model microorganisms do not give<br />
significant impacts on the other bacteria in pots <strong>and</strong> the greenhouse,<br />
although a long-term observation may be needed<br />
Keywords: Real-time PCR, , Horizontal gene transfer, Microbial community<br />
Effect of Fermentation Conditions on Biohydrogen Production<br />
from Lipid-rich Food Waste<br />
Soo Young MOON 1 , Hyo Sun KIM 1 , Can ZHANG 1 , Alma ABUG 1 ,<br />
Sung Chan CHOI 2 <strong>and</strong> Young Sook OH* 1<br />
1<br />
Dept. of Environmental <strong>Engineering</strong> <strong>and</strong> Biotechnology, Myongji University,<br />
Yongin 449-728, Republic of Korea. 2 J-47<br />
Dept. of Environmental Sciences <strong>and</strong><br />
Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea.<br />
*Corresponding author: ysoh@mju.ac.kr<br />
Although carbohydrate, protein, <strong>and</strong> lipid are basic components of<br />
organic material, hydrogen yield of carbohydrate fermentation has been<br />
reported to be significantly higher than that of protein <strong>and</strong> lipid. This<br />
study used lard as a model organic matter for lipid, <strong>and</strong> investigated its<br />
hydrogen production potential in batch anaerobic fermentation experiments<br />
under various combinations of stirring <strong>and</strong> CO2 scavenging condition to<br />
improve hydrogen production. To optimize hydrogen production, CO2<br />
scavenging was found to be more effective in lard fermentation compared<br />
to stirring. In the presence of both CO2 scavenging <strong>and</strong> stirring, the<br />
hydrogen yield of lard increased from 4.5 to 185.8 ml-H2/g-VS, <strong>and</strong> the<br />
hydrogen content of the total biogas was in the range of 87 - 93% (V/V).<br />
Acetic, propionic, butyric, <strong>and</strong> valeric acids were produced during the<br />
fermentation <strong>and</strong> valeric acid was found to be the major fatty acid (33.1<br />
- 67.1%) under all conditions. These results show that hydrogen production<br />
from lipid materials can be increased by minimizing the partial pressure<br />
of CO2 <strong>and</strong> hydrogen <strong>and</strong> activity of hydrogen-consuming bacteria.<br />
Keywords: hydrogen production, bioenergy, anaerobic fermentation<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
407
Inhibition of Quorum Sensing-Controlled Virulence Factors <strong>and</strong><br />
Biofilm Formation in Pseudomonas aeruginosa by<br />
2,5-Dimethyl-4-Hydroxy-3(2H)-Furanone (DMHF)<br />
Can ZHANG 1 , Yu-Hong JIA 1 , Soo-Young MOON 1 , Sung-Chan CHOI 2 <strong>and</strong><br />
Young-Sook OH* 1<br />
1<br />
Department of Environmental engineering <strong>and</strong> Biotechnology, Myongji<br />
University, Yongin 449-728, Korea. 2 J-48<br />
Department of Environmental Sciences <strong>and</strong><br />
Biotechnology, hallym University, Chunchon 200-702, Repubic of Korea.<br />
*Corresponding author: ysoh@mju.ac.kr<br />
2,5-Dimethyl-4-hydroxy-3(2H)-furanone (DMHF), an innoxious food<br />
additive, has been proved to have antimicrobial activity recently. However,<br />
few studies focus on its inhibitory effect on quorum sensing (QS) system<br />
in pathogenic bacteria. In this study, the production of N-(3-oxododecanoyl)-L-homoserine<br />
lactone (OdDHL) was completely inhibited by<br />
1.0 μM DMHF, while N-butyryl-L-homoserine lactone (BHL) production<br />
decreased on a dose dependent manner by DMHF. 1.0 μM DMHF also<br />
inhibited QS-controlled virulence factors such as rhamnolipid (55%<br />
decrease), lasA protease (40% decrease) <strong>and</strong> pyocyanin (52% decrease),<br />
<strong>and</strong> biofilm formation (83% decrease) of Pseudomonas aeruginosa PAO1<br />
without growth inhibition. However, the production of vi rulence factors<br />
was restored by the addition of exogenous OdDHL as well as BHL, which<br />
suggests the inhibition of DMHF is due to the interruption of QS with<br />
las system as the main target. But the specific inhibition mechanism of<br />
DMHF on QS system in Pseudomonas aeruginosa remains unknown. This<br />
study also gives evidence that OdDHL plays more significant role in the<br />
production of QS-controlled virulence factors than BHL.<br />
Keywords: Quorum sensing (QS), Biofilm formation, 2,<br />
5-Dimethyl-4-Hydroxy-3(2H)-Furanone (DMHF)<br />
The Effect of Treating Rice Straw with Lactobacillus plantarum<br />
BT-77 <strong>and</strong> Bacillus subtilis IN-55 on Silage Fermentation<br />
Hee Jong YANG 2,1 , Jae-Hwa HONG 1,3 , Seong-Yeop JEONG 1 ,<br />
Chan-sun PARK 1 , Hong-gi KIM 3 , Byung-Dae YOON 1 , Yeon-Woo RYU 2<br />
<strong>and</strong> Min-Soo KIM* 1<br />
1<br />
Bioindustrial Process Center, Korea Research Institute of Bioscience <strong>and</strong><br />
Biotechnology (KRIBB), Jeungeup 580-185, Korea. 2 Department of Molecular<br />
Science <strong>and</strong> Technology, Ajou University, san5, Wonchen-dong, Yeongtong-gu,<br />
Suwon, 443-749, Korea. 3 J-49<br />
Department of Agricultural Biology, Chungnam<br />
University, Daejeon 305-764, Korea.<br />
*Corresponding author: godfilts@kribb.re.kr<br />
This study aimed to the effects of Lactobacillus plantarum BT-77 <strong>and</strong><br />
Bacillus subtilis IN-55 on manufacturing of rice straw silage. We screened<br />
the strains having antibacterial <strong>and</strong> cellulase activity from traditional food<br />
<strong>and</strong> soil. Rice straw was treated with control (untreated; TGY broth media),<br />
BT-77 (4 × 105cfu/g; TGY broth media), IN-55 (4 × 105cfu/g; TGY<br />
broth media), BT-77 <strong>and</strong> IN-55 (4 × 105cfu/g; TGY broth media). Under<br />
optimal conditions (pH 5.5 <strong>and</strong> 30℃ for 7 days), treatments with BT-77<br />
increased the concentrations of lactic acid <strong>and</strong> improved the aerobic<br />
stability of rice straw. Moreover, BT-77 showed inhibitory activity against<br />
most of the indicator strains. Additionally, we confirmed not only high<br />
levels of a cellulase enzyme activity from BT-77 <strong>and</strong> IN-55, but also<br />
cell-wall degradation through an electron microscope. In this study, other<br />
treatments could improve silage quality, but mixed treatment was more<br />
effective as an additive for rice straw silage.<br />
Keywords: Rice straw silige, Inoculant, cellulase<br />
408 www.kormb.or.kr<br />
J-50<br />
Characterization of Manganese Peroxidase Isozyme H4 of<br />
Phanerochaete chrysosporium Expressed from the yeast Pichia<br />
pastoris<br />
Yu-Lim SON, Saravana Kumar KUMAR, Ae-Young MO, Hyun-Young KIM<br />
<strong>and</strong> Seung-Moon PARK*<br />
Division of Biotechnology, College of Environmental <strong>and</strong> Bioresource Sciences,<br />
Chonbuk National University, Jeonju, 561-756.<br />
*Corresponding author: smpark@chonbuk.ac.kr<br />
Lignin biodegradation is a key step for carbon recycling in terrestrial<br />
ecosystems, where white-rot basidiomycetes fungi degrade this recalcitrant<br />
wood polymer enabling cellulose utilization by microbial populations.<br />
Three peroxidases involved in lignin degradation are known as lignin<br />
peroxidase (LiP), manganese peroxidase (MnP), versatile peroxidase (VP).<br />
The cDNA encoding manganese peroxydase isozyme H4 (MnPH4),<br />
isolated from Phanerochaete chrysosporium, was cloned into the pPICZα<br />
vectors <strong>and</strong> expressed in Pichia pastoris under the control of the methanol<br />
inducible alcohol oxidase I (AOXI) promoter. Enzyme assay indicated that<br />
the recombinant MnPH4 is efficiently secreted into the medium upon<br />
hemoglobin supplementation, at a maximum concentration of 500 U l -1 .<br />
The purified recombinant manganese peroxidase (rMnP) exhibited a 60<br />
kDa molecular mass considerably larger, because of their glycosylation,<br />
<strong>and</strong> showed optimum pH of 4.5 <strong>and</strong> temperature at 35 o C.<br />
Keywords: Manganese Peroxidase, Phanerochaete chrysosporium, lignin<br />
peroxidase<br />
J-51<br />
Optimization of the Method to Nucleic Acids from Cyanobacteria<br />
in the Environmental Samples<br />
Hye-Ryoung KIM, Eun-Hye PARK, Kyoung-Hee OH <strong>and</strong><br />
Young-Cheol CHO*<br />
Dept. of Environmental <strong>Engineering</strong>, Chungbuk National University, Cheongju<br />
361-763, Korea.<br />
*Corresponding author: choy@cbnu.ac.kr<br />
In order to develop the efficient method to extract nucleic acids from<br />
cyanobacteria in the environmental samples, the concentration <strong>and</strong><br />
disruption methods for cyanobacteria were optimized. Three different<br />
kinds of filters, polycarbonate, cellulose nitrate, <strong>and</strong> GF/C, were used to<br />
concentrate cyanobacteria in the environmental samples. Cyanobacterial<br />
cells were treated with lysis buffer only or buffer combined with either<br />
of bead-beating or freezing-thawing. The extraction efficiency was<br />
examined through quantification of nucleic acids, <strong>and</strong> the purity was<br />
checked through PCR or RT-PCR. In case of DNA, the concentration<br />
with cellulose nitrate membrane <strong>and</strong> the disruption through bead-beating<br />
showed the highest extraction efficiency. On the other h<strong>and</strong>, the use of<br />
polycarbonate filter <strong>and</strong> freeze-thawing was adequate methods to extract<br />
RNA from cyanobacteria. It is needed to figure out why the extraction<br />
conditions for DNA <strong>and</strong> RNA were different. The methods developed<br />
in this study can be helpful to extract nucleic acids efficiently from the<br />
cyanobacteria in the natural aquatic environments. [Supported by grants<br />
from National Science Foundation of Korea]<br />
Keywords: Extraction, Cyanobacteria, Nucleic acid
Characterization of Four Soil Metagenome-Derived Lipolytic<br />
Enzymes Responsible for Hydrolysis of Autoinducer<br />
3-Hydroxypalmitic Acid Methyl Ester of Ralstonia solanacearum.<br />
Myung Hwan LEE 1 , Weixin TAO 2 , Nam Hee KIM 1 , Eul Chul HWANG 1 ,<br />
Jae Wook LEE 3 <strong>and</strong> Seon-Woo LEE* 1,2<br />
1<br />
Department of Applied Biology, Dong-A University, Busan 604-714, Republic<br />
of Korea. 2 Department of Medical Bioscience, Dong-A University, Busan 604-714,<br />
Republic of Korea. 3 J-52<br />
Department of Chemistry, Dong-A University, Busan<br />
604-714, Republic of Korea.<br />
*Corresponding author: seonlee@dau.ac.kr<br />
An autoinducer 3-hydroxypalmitic acid methyl ester (3-OH PAME) an<br />
unique quorum sensing signal molecule that regulates expression of<br />
virulence factors such as in phytopathogen Ralstonia solanacearum.<br />
Previously, we selected 4 metagenomic clones hydrolyzing 3-OH PAME<br />
among 142 142 lipolytic clones by using gas chromatography. All four<br />
clones were obtained from industrial waste contaminated soil metagenome<br />
<strong>and</strong> designated LP96, ULP104, ULP86 <strong>and</strong> UDL33. Lipolytic gene from<br />
LP96B, ULP104 <strong>and</strong> UDL33BB was over-expressed after cloning in<br />
pET-30b(+) in Escherichia coli BL21(DE3). The over-expressed, purified<br />
LP96, LP104 <strong>and</strong> Est33 exhibited hydrolysis activity toward 3-OH PAME.<br />
For determining of role of 3-OH PAME hydrolase in quorum sensing<br />
system of R. solanacearum GMI1000, pRKelp104 as pRK415 carrying<br />
lp104 gene was transformed into R. solanacearum GMI1000. Growth rate<br />
<strong>and</strong> exopolysaccharides (EPS) production of R. solanacearum GMI1000<br />
carrying pRKelp104 was slower <strong>and</strong> reducer than R. solanacearum<br />
GMI1000. These results showed that novel beta-hydroxypalmitic acid<br />
methyl ester hydrolases were discovered from industrial waste contaminated<br />
soil metagenome. The enzymes will interfere virulence factors<br />
expression in R. solanacearum by hydrolysis of autoinducer 3-OH PAME.<br />
Keywords: Metagenome, 3-OH PAME, beta-hydroxypalmitic acid methyl<br />
ester hydrolase<br />
Acute <strong>and</strong> Chronic Toxicity Assessment for Hazardous Effect<br />
of Caffeine to Daphnia magna<br />
Nam Hui HONG 2 , Thai-Hoang LE 2 , Sung-Kyu LEE 3 <strong>and</strong> Jiho MIN* 1<br />
1<br />
Graduate School of Semiconductor <strong>and</strong> Chemical <strong>Engineering</strong>, Chonbuk<br />
National University, 567, Baekje-Daero, Duckjin-gu, Jeonju, Jeonbuk, 561-756,<br />
KOREA. 2 Department of Bioprocess <strong>Engineering</strong>, Chonbuk National University,<br />
567, Baekje-Daero, Duckjin-gu, Jeonju, Jeonbuk, 561-756, KOREA. 3 J-53<br />
Toxicology<br />
Research Center, Korea Research Institute of Chemical Technology, Daejon,<br />
305-600, KOREA.<br />
*Corresponding author: hongnh@jbnu.co.kr<br />
Caffeine has been detected in water resulting in the chronic exposure<br />
of aquatic organisms. Because of organism exposed to hazardousness,<br />
aquatic ecosystems confusion is coming, <strong>and</strong> even it affect human. For<br />
this study, the impact was examined through the acute(48h) <strong>and</strong> chronic<br />
toxicity(21d) assay using Daphnia magna as animal model. In the acute<br />
toxicity test, 10 Daphnia neonates were exposed to several concentration<br />
of caffeine from 125㎍/l to 4000㎍/l. The lethal concentration with 50%<br />
mortability (LC50) was determined as about 2225㎍/l. In the chronic<br />
toxicity test, several small concentrations of caffeine such as 1%, 2%,<br />
10%, 20% LC50 were used to exposed organism for 21days. We found<br />
hazardous effect of caffeine on aquatic ecosystem. Although toxicity of<br />
caffeine was known to be week, we found that survival of Daphnia<br />
neonates was significantly decreased caused by toxicity of cafferine. Based<br />
on this result, it was found that aquatic ecosystems exposed by<br />
pharmaceutics can not be ignored.<br />
Keywords: Caffeine, Daphnia magna, Acute toxicity, Chronic toxicity.<br />
2011 International Symposium & Annual Meeting<br />
Natural Adaptation of Eubacterium limosum KIST612 for Biofuel<br />
Production<br />
Shinyoung PARK 1 , Daehee KIM 1 , Sunju CHOI 1 , In Geol CHOI 2 <strong>and</strong><br />
In Seop CHANG* 1<br />
1<br />
School of Environmental Science <strong>and</strong> <strong>Engineering</strong>, Gwangju Institute of Science<br />
<strong>and</strong> Technology (GIST), 261 Cheomdan-gwagiro, Buk-gu, Gwangju 500-712,<br />
Korea. 2 J-54<br />
College of Life Sciences <strong>and</strong> Biotechnology, Korea University,<br />
Anam-Dong, Seongbuk-Gu, Seoul,136-713, Korea.<br />
*Corresponding author: ischang@gist.ac.kr<br />
The production of chemicals by microbial reaction has become an<br />
increasingly alternative to current petroleum-based chemical synthesis.<br />
Eubacterium limosum KIST612 has been the organism of choice for the<br />
biochemicals production form hydrogen or carbon monoxide. E. limosum<br />
KIST612 produces (1) organic acids - acetic <strong>and</strong> butyric acids, (2) solventethanol<br />
<strong>and</strong> (3) gas- carbon dioxide (CO2). Among them, acetate <strong>and</strong><br />
butyrate are major products. We hypothesized that constant transfer in<br />
certain medium condition this microbe may try to adapt to the given<br />
environmental condition. We hypothesized that if an excess of acetate<br />
or butyrate exist with CO as a main electron source, surplus electron<br />
may be utilized for production of more reduced products.Therefore, the<br />
hypothesis of this experiment is that excessive acetate <strong>and</strong> butyrate<br />
addition into the medium could change metabolic flux. We have tested<br />
the effect of acetate or butyrate addition for longer than 1 year with CO<br />
as a main energy source. We found significant increase of solvents, ethanol<br />
<strong>and</strong> butanol production after 1 year adaptation. It was also clearly observed<br />
that acetate could be converted to more reduced products in the presence<br />
of excess CO.<br />
Keywords: Acetogen, Eubacterium, Adaptation<br />
J-55<br />
Mortality of Moth Larvae by Various Entomopathogenic Fungi<br />
Young-Hoon LIM, Eun-Soo GO <strong>and</strong> Keun KIM*<br />
Dept. of Bioscience <strong>and</strong> Biotechnology, The University of Suwon, Gyeonggi-do<br />
445-743, Korea.<br />
*Corresponding author: limyounghoon@korea.com<br />
To determine the virulence of the various fungi, 10 8 conidia/㎖<br />
suspension of each fungal strain was sprayed onto 2rd instar of larvae<br />
on a dampened filter paper in a petri-dish <strong>and</strong> the petri-dish was incubated<br />
at 25℃ for 15 days. The mortality by the different fungal strains varied<br />
from 20% to 100%. Among various fungi tested, 4 strains showed relativity<br />
high mortality against larvae. The mortality of the larvae increased <strong>and</strong><br />
the 50% lethal time(LT50) decreased significantly as the applied conidial<br />
concentration was raised. LT50 values of strain No. 198, 216, 218, <strong>and</strong><br />
500 against larvae were 12.7, 9.5, 8.4, <strong>and</strong> 9.3 days, respectively. Mean<br />
lethal concentration(LC50) values of strain No. 198, 216, 218 <strong>and</strong> 500<br />
against the larvae were 3.62 × 10 7 , 9.94 × 10 6 , 6.97 × 10 4 , <strong>and</strong> 3.01<br />
× 10 5 conidia/㎖, respectively.<br />
Keywords: Entomopathogenic Fungi, Mortality, Moth Larvae<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
409
J-56<br />
Development of Baeyer-Villiger Monooxygenases Screening<br />
Protocols Using High-throughput screening (HTS) system<br />
Gowoun LEE, Yunjon HAN, Jae Jun SONG <strong>and</strong> Jong Hyun CHOI*<br />
Enzyme-based Fusion Technology Research Team, Microbe-based Fusion<br />
Technology Research Center, Korea Research Institute of Bioscience <strong>and</strong><br />
Biotechnology (KRIBB), 1404 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea.<br />
*Corresponding author: jhchoi@kribb.re.kr<br />
Baeyer-Villiger Monooxygenases (BVMOs) are flavoenzymes that<br />
catalyze the insertion of an oxygen atom in a C-C bond using NADPH<br />
<strong>and</strong> allow the conversion of (cyclic)-ketones into lactones or esters. For<br />
the development of BVMO screening protocol, we used fluorescence based<br />
substrate <strong>and</strong> robot based HTS system. Optimized BVMO screening<br />
protocol for high throughput screening system led to 6,000 clone active<br />
verification per a day was available. As result, we screened the new type<br />
of BVMO positive clone which is named as “GomsoMF” from "Gomso"<br />
metagenomic library. This “GomsoMF” was novel BVMO like enzyme<br />
can convert cyclohexanone to ε-caprolactone. Also this enzyme was<br />
convert cyclopentanone <strong>and</strong> cycloheptanone into δ-valerolactone <strong>and</strong><br />
7-heptanolactone respectively. This new BVMO will be used for the<br />
synthesis of useful biochemicals or the biotechnological application. (This<br />
work was supported by a high throughput screening (HTS)-based<br />
Integrated Technology Development grant from the MEST, <strong>and</strong> a basic<br />
research grant from KRIBB.)<br />
Keywords: Baeyer-Villiger Monooxygenases (BVMOs), high throughput<br />
screening<br />
J-57<br />
The Effect of Gamma Irradiation on Poliovirus, a Model for<br />
Norovirus inoculated in Oyster (Crassostrea Gigas) <strong>and</strong> Effect<br />
of Culture Condition<br />
Jong-Il CHOI, Eu-Ri JO, Pil-Mun JUNG, Jae-Hun KIM, Beom-Suk SONG,<br />
Jae-Kyung KIM, Jong-Heum PARK <strong>and</strong> Ju-Woon LEE*<br />
Team for Radiation Food Science <strong>and</strong> Biotechnology,Advanced Radiation<br />
Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185.<br />
*Corresponding author: sjwlee@kaeri.re.kr<br />
Poliovirus is a recognized surrogate for norovirus, pathogen in water<br />
<strong>and</strong> food, due to the structural <strong>and</strong> genetic similarity. Although radiation<br />
sensitivity of poliovirus in water or media had been reported, there has<br />
been no research in food model such as shellfish. In this study, oyster<br />
(Crassostrea gigas) was incubated in artificial seawater contaminated with<br />
poliovirus, <strong>and</strong> thus radiation sensitivity of poliovirus was determined in<br />
inoculated oyster. The effects of ionizing radiation on the sensitivity of<br />
poliovirus were also evaluated under different conditions such as pH (4-7)<br />
<strong>and</strong> salt concentration (1-15%) in culture broth, <strong>and</strong> temperature during<br />
irradiation. The D10 value of poliovirus in PBS buffer, virus culture broth<br />
<strong>and</strong> oyster was determined to 0.46, 2.84 <strong>and</strong> 2.94 kGy, respectively. The<br />
initial plaque forming unit (PFU) of poliovirus in culture broth was slightly<br />
decreased as the decrease of pH <strong>and</strong> the increase of salt concentration,<br />
but radiation sensitivity was not affected by pH <strong>and</strong> salt contents. However,<br />
radiation resistance of poliovirus was increased at frozen state. These<br />
results provide the basic information for the inactivation of pathogenic<br />
virus in foods by using irradiation.<br />
Keywords: Radiation sensitivity, Poliovirus, Oyster<br />
410 www.kormb.or.kr<br />
Application of Bacillus subtilis 168 to Improve Durability of<br />
Cement<br />
Sung-jin PARK 1 , Jong-Myong PARK 1 , Wha-Jung KIM 2 <strong>and</strong><br />
Sa-Youl GHIM* 1<br />
J-58<br />
1<br />
Dep. of <strong>Microbiology</strong>, Kyungpook National University, Daegu 702-701, Korea.<br />
2<br />
Dept. of Architecture <strong>Engineering</strong>, Kyungpook National University, Daegu<br />
702-701, Korea.<br />
*Corresponding author: ghimsa@knu.ac.kr<br />
Calcium carbonate precipitation, a widespread phenomenon among<br />
bacteria, has been investigated due to its wide range of scientific <strong>and</strong><br />
technological application in concrete remediation. In this study, Bacillus<br />
subtilis 168 which could induce calcium carbonate precipitation, was used<br />
to coat on the cement surface. CaCO3 crystal aggregates <strong>and</strong> organic matrix<br />
molecules of these strains were deposited on the surface of the cement<br />
paste. To verify the effect of coating on the surface of the cement paste,<br />
water permeability of the paste was measured by water immersion test.<br />
In the results, B. subtilis 168 addition showed lower water permeability<br />
than those treated with B4 medium only, suggesting that the precipitated<br />
calcium induced by B. subtilis 168 could be used as a sealing agent to<br />
block water penetration. Also, the calcium precipitation on the surface<br />
of cement mortar could increase the compressive strength of the mortar.<br />
The compressive strength test showed that the cement mortar treated with<br />
B. subtilis 168 showed an improvement of compressive strength.<br />
Keywords: Calcium carbonate precipitation, Bacillus subtilis 168, cement<br />
J-59<br />
Screening Enzyme from Single cell Based Polymerase Fosmid<br />
Cloning<br />
Yunjon HAN, Kyong-Cheol KO, Jong Hyun CHOI <strong>and</strong> Jae Jun SONG*<br />
Enzyme-based Fusion Technology Research Team, Microbe-based Fusion<br />
Technology Research Center, Korea Research Institute of Bioscience <strong>and</strong><br />
Biotechnology (KRIBB), 1404 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea.<br />
*Corresponding author: jjsong@kribb.re.kr<br />
"Metagenomics," which is the genomic analysis of the microbial<br />
community attempts to overcome culturability of various microbes. In<br />
general, the diversity of metagenomic libraries was affected by abundance<br />
of bacteria in environmental sample. Therefore the research of minor<br />
microbial texa in environmental samples remains the biggest challenge.<br />
A new method was developed for enrichment minor bacteria from<br />
environmental samples. This method is based on the Fluorescence in situ<br />
hybridization(FISH), Fluorescence associated cell sorter(FACS) <strong>and</strong><br />
Multiple displacement amplification(MDA). We demonstrated enrichment<br />
minor bacteria from artificial microbial community <strong>and</strong> single cell based<br />
MDA followed by fosmid library construction for enzyme screening. The<br />
Jeongupia naejangsanesis in which the cellulolytic activity is known was<br />
used in the minor microorganism of the artificial microbial community.<br />
As a result, we obtained MDA products from single bacterial cell. And<br />
we could generate fosmid library having an average insert size of 32<br />
kb. Among this fosmid library we screened two new cellulose genes. (This<br />
work was supported by a high throughput screening (HTS)-based<br />
Integrated Technology Development grant from the MEST, <strong>and</strong> a basic<br />
research grant from KRIBB.)<br />
Keywords: Single cell, Multiple displacement amplification, Screening<br />
enzyme
Virulence of Entomopathogenic Fungus Lecanicilium sp. Le85<br />
towards Cotton Aphid<br />
Van Hanh VU* 1 , Dinh Thi QUYEN 2 , Xuan Dat VU 2 , Huu Quan NGUYEN 2<br />
<strong>and</strong> Keun KIM 3<br />
1<br />
Institute of Biotechnology, Vietnam Academy of Science <strong>and</strong> Technology, Hanoi<br />
10600, Vietnam/ Department of Bioscience <strong>and</strong> Biotechnology, The University of<br />
Suwon, Hwaseong-si 445-743, Korea. 2 Institute of Biotechnology, Vietnam<br />
Academy of Science <strong>and</strong> Technology, Hanoi 10600, Vietnam. 3 J-60<br />
Department of<br />
Bioscience <strong>and</strong> Biotechnology, The University of Suwon, Hwaseong-si 445-743,<br />
Korea.<br />
*Corresponding author: vvhanh@ibt.ac.vn<br />
Twenty-two entomopathogenic fungal strains were used for laboratory<br />
bioassay against aphid Aphis gossypii living on cucumber leaves. Among<br />
them, four strains of Lecanicillium spp. isolated from infected aphid’s<br />
cadavers had high virulence towards A. gossypii causing 84 to 100%<br />
mortality after 5 to 7 days of conidial treatment. Especially, one fungus<br />
Lecanicillium sp. Le85 showed the highest virulence towards development<br />
<strong>and</strong> reproduction of A. gossypii. At 20~24 o C <strong>and</strong> relative humidity of<br />
75~85%, after 5 days of application with 1x10 8 conidia of Le85 /ml, A.<br />
gossypii did not develop <strong>and</strong> exhibited a mortality of 100% with LT50<br />
of 2.6 days. Moreover, at 25~31 o C <strong>and</strong> relative humidity of 60~70%,<br />
the mortality of A. gossypii by Le85 was about 95 ~100% after 5 days<br />
with LT50 of 2.8 days. These results suggested the potential role of Le85<br />
for use as microbial control agent against A. gossypii in greenhouses.<br />
Keywords: Cotton aphids Aphis gossypii, entomopathogenic fungi,<br />
Lecanicillium sp.<br />
J-61<br />
Partial Identification of Microorganisms from Artificially<br />
Constructed Tunnel <strong>and</strong> Dokdo Soil to Develop Antifungal<br />
Construction Material<br />
Jong-Myong PARK, Sung-Jin PARK <strong>and</strong> Sa-Youl GHIM*<br />
School of Life Sciences, Kyungpook National University, Daegu 702-701, Korea.<br />
*Corresponding author: eveningwater@hanmail.net<br />
To develop antifungal concrete, we sampled microorganisms from the<br />
tunnel to determine the most widely distributed fungal strain. In addition,<br />
we are finding the multi-functional (antifungal <strong>and</strong> calcite forming)<br />
bacterium from the Dokdo soil. Isolation <strong>and</strong> pure culture of bacterium<br />
<strong>and</strong> fungi were done using potato dextrose agar <strong>and</strong> several bacteriological<br />
media.As the result of partial identification, fungal strain showed the<br />
closest homologies with Penicillium sp., Tritirachium sp., Elsinoaceae sp.,<br />
Cladosporium spp., Alternaria spp., Pestalotiopsis cocculi, Pseudozyme<br />
prolifica or Aureobasidium pullulans, respectively. Each bacterial strains<br />
showed the closest similarity with Rhodococcus erythropolis (99.794%),<br />
Bacillus aryabhattii (99.791%) or Bulkholderia glathei (96.369%),<br />
respectively. The most widely distributed fungal strain was Cladosporium<br />
spp. As the result of the calcite forming assay, 52 Dokdo derived bacterial<br />
strains <strong>and</strong> five tunnel derived strains were identified as calcite forming<br />
bacteria (CFB). Qualitative antimycotic assay showed that 23 Dokdo <strong>and</strong><br />
two tunnel derived CFB strains have antimycotic activity against<br />
Aspergillus niger KCTC6906 <strong>and</strong> Cladosporium spp. using the strain's<br />
crude liquid culture.<br />
Keywords: Antifungal, Construction, Material.<br />
2011 International Symposium & Annual Meeting<br />
Novel Esterases Isolated from Marine Sediment Metagenome<br />
So-Hyeon SEO 1 , Sang-Hong YOON 1 , Young-Seok LEE 1 ,<br />
Sung-Gyun KANG 2 , Jung-Hyun LEE 2 <strong>and</strong> Chang-Muk LEE* 1<br />
1<br />
Functional Biomaterial Division, National Academy of Agricultural Science,<br />
Rural Development Administration, Suwon 441-707, Korea. 2 J-62<br />
Korea Ocean<br />
Research <strong>and</strong> Development Institute, Ansan 426-744, Korea.<br />
*Corresponding author: changmuk@rda.go.kr<br />
Metagenomes from uncultured microbial communities are rich sources<br />
for identifying novel biocatalysts. With deep sea sediments at an isl<strong>and</strong>,<br />
Dokdo, we have constructed the metagenomic libraries comprising total<br />
60,672 fosmid clones. A fosmid clone with lipolytic activity was selected<br />
by functional screening of the metagenomic libraries using tributyrin agar<br />
plates. To identify genes, a shot-gun library was constructed from the<br />
selected clone by r<strong>and</strong>om shearing. Further functional screening of the<br />
shot-gun library revealed one novel clones. The sequence analysis of the<br />
clone revealed the presence of an open reading frame encoding acidic<br />
polypeptide of 59.4kDa (553 amino acids, pI of 4.45) with the highest<br />
homology to a carboxyl esterase from δ-proteobacterium Haliagium<br />
ochaceum DSM 14365(44% identity, 61% similarity). The sequence<br />
analysis showed that the protein harbors the typical catalytic triad formed<br />
by Ser229, Asp347 <strong>and</strong> His461. Multiple sequence alignments with known<br />
carboxyl esterases showed the clone belongs to the carboxylesterase BioH<br />
member, which is involved in biotin biosynthesis in Aromatoleum<br />
aromaticum [This experiment is supported by grants from the Rural<br />
Development Administration project number PJ0067032011 <strong>and</strong> IPET<br />
project number 110037-03-1-HD110].<br />
Keywords: carboxylesterase, metagenome, dokdo<br />
K_ Host-Microbe Interactions<br />
K-1<br />
Aphids Feeding Manipulates Host Plant Immunity against Plant<br />
Pathogens<br />
Choong-Min RYU* <strong>and</strong> Soohyun LEE<br />
Laboraytory of Bio-industry <strong>and</strong> Biochemistry Research Center, KRIBB 111<br />
Gwahangno, Yuseong-gu, Daejeon 305-806, S. Korea.<br />
*Corresponding author: cmryu@kribb.re.kr<br />
Plants modulate defense signaling networks to response against different<br />
biotic stresses. In this work, we evaluated whether insect infestation can<br />
change of plant defense mechanism charging to subsequent pathogen<br />
attack. Aphids <strong>and</strong>/or a chemical trigger benzothiadiazol (BTH) was<br />
pretreated seven days before challenging two pathogenic bacteria,<br />
Xanthomonas axonopodis pv. vesicatoria (Xav) as a compatible pathogen<br />
<strong>and</strong> X. axonopodis pv. glycines 8ra (Xag 8ra) as an incompatible pathogen.<br />
Disease severity was noticeably lower in aphid <strong>and</strong> BTH + aphid<br />
treatments than control. BTH or aphids alone treatment did not differ<br />
induction of hypersensitive response (HR) against Xag 8ra. However, the<br />
combination treatment showed synergistically to reduced HR appearance.<br />
Pretreatment of BTH resulted in significant reduction of aphids population.<br />
Expression of the two defense-related genes, Capsicum annum<br />
Pathogenesis-related genes 1 (CaPR1) <strong>and</strong> CaPR4 revealed that aphid<br />
infestation primes resistance genes resulting in induction of systemic<br />
defense response against compatible <strong>and</strong> incompatible pathogens. Taken<br />
together, our results suggest that insect feeding itself elicits propound plant<br />
resistance <strong>and</strong> help plant coping subsequent attack of pathogens.<br />
Keywords: Aphid, immunity, Xanthomonas<br />
The Korean Society for <strong>Microbiology</strong> <strong>and</strong> Biotechnology<br />
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