J_Environmetal Microbiology and Engineering

Isolation and Characterization of Lactobacillus Pentosus Kimchi 

from the Fermented Korean Food, Cutlassfish Kimchi 

Ji-Hyeong KIM 1 , Mi-Sun KWAK 1 , Hyeon-Jun SON 1 , Se-Jeung JANG 2 , 

Jong-Bok RHO 3 , Young-Ki CHOI 4 and Moon-Hee SUNG* 1 

1 

Dept. of Advanced Fermentation Fusion Science &Technology, Kookmin 

University, Korea. 2 Dept. Bio &Nanochemistry, Kookmin University, Seoul, 

Korea. 3 BioLeaders Corporation, Daejeon, Republic of Korea. 4 I-101 

College of 

Medicine &Medical Research Institute, Chungbuk National University, Korea. 

*Corresponding author: safe2112@hanmail.net 

Kimchi, a Korean traditional fermented food, is prepared with various 

vegetables such as cabbage, radishes, spices, and other seasonings. The 

fermentation of kimchi is carried out by microorganisms, mainly GRAS 

lactic acid bacteria (GRAS-LAB). Recently, several health functions of 

kimchi have been reported, including antimicrobial effects and anti-viral 

effects. In order to isolate kimchi LAB, which has antimicrobial and 

anti-viral effects, we screened the kimchi LAB from cutlassfish kimchi. 

We isolated kimchi LAB that could grow in an MRS plate medium 

containing 2% CaCO3. Finally, one strain was selected for the protease 

activity on a 1% milk plate medium. The isolated strain was identified 

according to its phenotypic characteristics and the data from the 

phylogenetic sequence analysis of the 16S rRNA gene. We examined 

antimicrobial activity, antifungal activity, and anti-viral activity with 

cell-free supernatants and a culture broth of kimchi LAB grown in MRS 

broth. [This work was supported by the Seoul R&BD Program (10580) 

and Top Brand Project grant from Korea Research Council of Fundamental 

Science &Technology (KGM3110912).] 

Keywords: fermentation, Kimchi, lactobacillus 

J _ Environmetal Microbiology and Engineering 

J-1 

Growth Promoting Effect of Alginate on Nannochloropsis oculata 

Hyun-Jin PARK 1 , Young-Hwa KIM 2 and Jae-Hwa LEE* 1 

1 Department of Bioscience and Biotechnology, College of Medical and Life 

Science, Silla University, Busan 617-736, Korea. 2 Department of Pharmaceutical 

Engineering, College of Medical and Life Science, Silla University, Busan 

617-736, Korea. 

*Corresponding author: phj0386@naver.com 

The growth of Nannochloropsis oculata (N. oculata), unicellular 

microalgae, by alginate was investigated. Alginate was depolymerized with 

sulfuric acid (H2SO4) and heat (121℃), simultaneously. Addition of 0.75% 

alginate oligomer depolymerized with 0.2 N H2SO4 showed the maximum 

yield and the growth rate of N. oculata. Chlorophyll content and reducing 

sugar was increased by alginate oligomer in a dose-dependent manner. 

Alginate oligomer promoted the growth of N. oculata, whereas the original 

alginate polysaccharides had no significant effect. Laminaria japonica (L. 

japonica) extract containing high level of alginate was also increased 

growth rate and chlorophyll content. CO2 supply addition to L. japonica 

extract showed no change the growth rate, although addition to alginate 

oligomer showed prominently increased. N. oculata could use more 

saccharides in presence of CO2 according to reducing sugar determination. 

From these results, it is useful to establish optimal condition for high 

cell density cultivation of N. oculata. 

Keywords: Alginate, Nannochloropsis oculata, Laminaria japonica 

396 www.kormb.or.kr 

Molecular Monitoring of Microbes in Surface water and 

Distribution System of Pipeline using PCR-DGGE 

Hee Suk LEE* 1 , Sun-Bok GEUN 2 and Min Jeoung KIM 1 

J-2 

1 2 

K-water. KOTTI. 

*Corresponding author: lovealga@kwater.or.kr 

The objective of this study is to monitor the microorganisms in various 

surface water and distribution system of pipeline in the use of Denaturation 

Gradient Gel Electrophoresis (DGGE) system. Five major surface water 

(GR, DH, BS, IS, CJ) and two samples of distribution system (DB, US) 

in the pipeline were used. Nucleic acids of these samples were amplified 

using PCR and confirmed by DGGE using 8% polyacryamide gel. The 

microbial communities of samples which were taken in November were 

more diversified than in August. The microbial community in the 

distribution system was also observed and compared with the data from 

surface water. The diversity of microbial community in the distribution 

system is less than that of surface wate. The PCR-DGGE methods can 

be used as a useful tool to compare the microbial community and expect 

the variation of microbial community. 

Keywords: Monitoring, PCR-DGGE, Surface water 

J-3 

Growth Kinetics of Algae under the Influence of Antibiotics 

using Toxy-PAM 

Man-Gi CHO*, Pina KNAUFF, Katharina KRÄHLING, Thomas AMBROSI, 

Vera KEIL, Khalida MANSOURI, Daniel ASCHENBRENNER, 

Binod PRASAD and Thiyam GENERAL 

Department of Bio-Chemical Engineering, Dongseo University Graduate 

School,Busan 617-716, Republic of Korea. 

*Corresponding author: mgcho@gdsu.dongseo.ac.kr 

With increasing importance of micro algae in the pharmacy, medicine, 

environmental engineering and further fields, it is necessary to investigate 

growth conditions and potential growth inhibitors. Genetical modifications 

of micro algae strains can be an alternative to meet the increasing demand 

of valuable compounds from micro algae. Thus the following work with 

respect to suspectibility of micro algae Tetraselmis suecica, Marine 

Chlorella sp. and Haematococcus pluvialis against antibiotic was carried 

out to investigated the growth kinetics under influence of antibiotics. 

Different concentration were applied and the strains were incubated for 

7 days. The cell number, the optical density (OD) and the fluorescence 

activity (F0, Fm, Y) using Toxy PAM were measured.The cell number 

and the optical density are continuously increasing with the time. The 

fluorescence activity appears independent of the antibiotic concentration. 

Keywords: Microalgae, Antibiotics and Fluorescence, Growth Kinetic

Peptide Self-assembled Simultaneous Formation of 

Organic-inorganic Hybrid Spheres and Their Potential 

Applications 

Jungok KIM 1 and Hor-Gil HUR* 1,2 

1 

School of Environmental Science and Engineering, Gwangju Institute of Science 

and Technology, Gwangju, Korea. 2 J-4 

International Environmental Research Center, 

Gwangju Institute of Science and Tehnology, Gwangju, Korea. 

*Corresponding author: kjungok@gist.ac.kr 

The hybrid structures composed of organics and inorganic nanoparticles 

has been attractive due to their improved properties and stabilities 

compared to homogeneous materials. Despite some advances, there are 

still limited works on the simultaneous assembly of biological molecules 

(e.g. peptide) and inorganic nanoparticles into well-designed 

supramolecular structures under aqueous conditions. In this study, hybrid 

spheres containing peptides and gold nanoparticles has been 

simultaneously synthesized in water using AG4 peptides which acted as 

a reducing agent to guide the nucleation and growth of gold nanoparticles 

and a precursor to form sphere-like structure by self-assembly where the 

size of hybrid spheres is precisely controlled by adjusting the operating 

temperature. The self-assembled peptide spheres remain stable even after 

selective removal of the gold nanoparticles by iodide etching. The surface 

of gold nanoparticles has been functionalized using thiol group linked 

to biomolecules. The ability to synthesize nanoparticle and self-assembled 

peptide structures with controlled size and composition in an 

environmental benign way will allow fabricating new class of 

multi-functional organic-inorganic hybrid superstructures for various 

biomedical and electronic applications. 

Keywords: peptide, self-assembly, gold nanoparticle 

Enhancement of Biomass and Fatty acids of Dunaliella salina 

According to the Culture Medium Compositions Containing the 

Animal Hormone, Steroids 

Mi-Kyung KIM* 1 , Yun-Sam KIM 2 , Dong-Mok LEE 3 and Inho CHOI 3 

1 

EcoPhycoTech, Marine Science Research Center, Yeungnam University, 

Gyeongsan 712-749, Korea. 2 Dept. of Biology, Yeungnam University, Gyeongsan 

712-749, Korea. 3 J-5 

School of Biotechnology, Yeungnam University, Gyeongsan 

712-749, Korea. 

*Corresponding author: mkkim@yu.ac.kr 

The main issues are focused on how to enhance efficiently the biomass 

for algal industrialization as a future bioresource: supplemental nutrients, 

pharmaceutical products, bioremediation, biofuel and reduction of 

greenhouse gas (CO2) emissions. The extracts of the wasted pig-testis 

(PTSE) contain steroid hormones - nondrolone (55%), testosterone (28%), 

androstenedione (1%), estradiol (5%), estrone (4%), progesterone (7%) 

or the ones of pig-placenta (PPSE) containing nondrolone (13%), 

testosterone (0.2%), androstenedione (1.7%), estradiol (2%), estrone 

(16.5%) and progesterone (66.6%), respectively. These extracts were 

added to the D media according to the different ratios of 5 ng/ml, 50 

ng/ml, 500 ng/ml for PTSE, 10 ng/ml, 100 ng/ml, 1 ug/ml for PPSE, 

respectively. The additions of PTSE of 5 ng/ml and PPSE of 10 ng/ml 

increased the cells densities of 250% and 200% , while the PTSE of 

500 ng/ml and PPSE of 1 ug/ml increased the fatty acids of 5500% and 

8260% for Dunaliella salina, respectively. The amount of DHA(C22:6) 

of D. salina was two-fold more accumulated in PTSE of 500 ng/ml (22.3%) 

and PPSE of 1 ug/ml (24.4%) than control D medium. The mechanism 

of gene expression by animal steroids should be clarified in D. salina 

cultured in the different hormone compositions. 

Keywords: Steroid, Fatty acids, Dunaliella salina 

2011 International Symposium & Annual Meeting 

Effect of Biosurfactant-Producing Bacterium on Mineralization 

of Wastewater Contaminated with Petroleum by Petroleum- 

Degrading Bacterial Community 

Bo Young JEON 1 , Jun-Young YI 1 , Il Lae JUNG 2 and Doo Hyun PARK* 1 

1 

Department of Chemical and Biological Engineering, Seokyeong University, 

Seoul 136-704, Korea. 2 J-6 

Department of Radiation Biology, Environmental 

Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon 

305-353, Korea. 

*Corresponding author: baakdoo@skuniv.ac.kr 

Serratia sp. was isolated from riverbed soil in Jungrang-cheon and 

cultivated with a mineral-based soybean oil medium. Serratia sp. grew 

in the mineral-based soybean oil medium but did not grow in a 

mineral-based kerosene-diesel medium. The petroleum-degrading bacteria 

used in this study--Acinetobacter sp., Pseudomonas sp., Paracoccus sp., 

and Cupriavidus sp.--were isolated from a specially designed enrichment 

culture. The mineralization efficiency of wastewater contaminated with 

kerosene and diesel (WKD) by the PDBC was increased significantly by 

addition of the crude biosurfactant. The mineralization efficiency of the 

WKD was also enhanced by co-culture of Serratia sp. and PDBC. The 

temperature gradient gel electrophoresis (TGGE) pattern for the 16S rDNA 

variable region extracted from the co-cultured Serratia sp. and PDBC 

in the WKD at 2 nd day of incubation time was identical to that at the 

8 th day of incubation. In conclusion, the co-culture of Serratia sp. and 

PDBC is an applicable technique for the mineralization of wastewater 

contaminated with petroleum, which may substitute for a biosurfactant. 

Acknowledgement This work was supported by the New &Renewable 

Energy of the KETEP grant funded by the Korea governmental Ministry 

of Knowledge Economy (2010T1001100334) 

Keywords: Biosurfactant, Serratia sp. petroleum-degrading bacteria, mixed 

culture, TGGE 

J-7 

Effect of Electrochemical Pulse on Microbial Community 

Variation and Growth of Crops 

Doo Hyun PARK, Jun-Young YI and Ji-Won CHOI* 

Department of Biological Engineering, Seokyeong University, Seoul 136-704, 

Korea. 


Electrochemical pulse generated from DC 2, 4, 6, 8, 10 V of electricity 

was induced to farm soil, in which lettuces were cultivated. The pulse 

intervals were controlled by exchange of electrode polarity at intervals 

of 10s. Distance between electrodes was adjusted to 10 cm, by which 

the unit of electric intensity charged to the farm soil was 200, 400, 600, 

800, and 1,000 mV per cm, respectively. Bacterial community diversity 

was analyzed and compared by temperature gradient gel electrophoresis 

and species identity of the bacterial community was determined based 

on the sequence homology of 16S-rDNA variable region that was extracted 

from TGGE gel. Lettuce growth was not activated in the electrochemical 

pulsed condition at 200 and 400 mV/cm of electrochemical potential but 

activated at 600, 800, and 1,000 mV of electrochemical potential in 

proportion to the electric intensity. Acknowledgement This research was 

supported by Seokyeong University in 2010 

Keywords: electrochemical pulse, bacterial community, soil bacteria 

The Korean Society for Microbiology and Biotechnology 

397

Characterization of Phosphate-Solubilizing Bacteria Isolated from 

Soil 

Ah Ra CHO 1 , Jae Gon KANG 2 , Hoon Seok KANG 2 and Dae Ook KANG* 1 

1 

Department of Biochemistry and Health Science, Changwon National University, 

Changwon 641-773, Korea. 2 J-8 

Korea Biochemical Inc., Igok-ri, Munsan-eup, Jinju 

660-841, Korea. 

*Corresponding author: biochem1@changwon.ac.kr 

Bacteria capable of forming halos on Pikovskaya's agar medium were 

isolated from soil as an effort to develop microbial agent for the amendment 

of inorganic phosphate-accumulated soil. Five bacterial isolates were 

finally selected and identified using biochemical and molecular taxonomy. 

Four isolates were identified as Acinetobacter calcoaceticus and the one 

Pantoea sp. Phosphate-solubization activity gradually increased for 7-day 

growth, which was verified by reciprocal decrease in elctronic conductance. 

The optimal temperature and initial pH were 30 o C and 5.0 - 6.0, 

respectively. The addition of 3% of NaCl had no effect on cell growth. 

We selected optimal C-, N-source and inorganic salts. The Pantoea sp. 

showed 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. We 

examined 2D protein expression profies of three isolates between normal 

and salt stressed (4% NaCl) culture condition. 

Keywords: ACC deaminase, electronic conductance, optimal temperature, 

pH, phosphate-solubilizing bacteria, 

Laboratory Evaluation of Entomopathogenic Fungi for Control 

of Green Peach Aphid Myzus persicae 

Van Hanh VU* 1 , Thi Thuy Duong LE 2 , Dinh Thi QUYEN 2 and Keun KIM 3 

1 

Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi 

10600, Vietnam/ Department of Bioscience and Biotechnology, The University of 

Suwon, Hwaseong-si 445-743, Korea. 2 Institute of Biotechnology, Vietnam 

Academy of Science and Technology, Hanoi 10600, Vietnam. 3 J-9 

Department of 

Bioscience and Biotechnology, The University of Suwon, Hwaseong-si 445-743, 

Korea. 

*Corresponding author: vvhanh@ibt.ac.vn 

Twenty seven entomopathogenic fungal strains were isolated from 

various infected aphids. One fungus exhibited the highest virulence 

towards green peach aphids, Myzus persicae. The isolate conformed most 

closely to Lecanicillium sp. based on the sequence of internal transcribed 

spacer region of its 28S rDNA, and thus was designated as Lecanicillium 

sp. Le85. The laboratory bioassay for the aerial conidia of Le85 was 

performed for the control of M. persicae. After six days of application 

with 1×10 8 aerial conidia/ml, at 20~25 o C and relative humidity of 75~85%, 

the mortality of aphid by Le85 was about 97% with LT50 of 3.2 days. 

Meanwhile, at 26~31 o C and relative humidity of 60~70%, the evaluations 

of Le85 on potted cabbages showed an aphid mortality ranging from 82 

to 95%. Significant conidiation was observed on the aphid cadavers 

infected with Le85. Undoubtedly, the results suggest that Le85 can be 

used as a biocontrol agent for aphid M. persicae on cabbages in 

greenhouses. 

Keywords: Myzus persicae, entomopathogenic fungi, Lecanicillium sp., 


Glucose Utilization and Electricity Generation Using Genetically 

Modified Shewanella Strains 

DongGeon CHOI 1 , Sae Bom LEE 1 , In Geol CHOI 2 and In Seop CHANG* 1 

1 


and Technology(GIST), 261 Cheomdan gwagi-ro, Buk-gu, Gwangju 500-712, 

Korea. 2 J-10 

College of Life Science and Biotechnology, Korea University, Anam-dong, 

Sungbuk-gu, Seoul 136-701, Korea. 

*Corresponding author: ischang@gist.ac.kr 

Genomic sequence analyses of Shewanella strains have been studied 

for carbon metabolism as well as their physiological characteristics. Even 

though Shewanella strains seem to have glycolysis pathways and enzymes 

as central carbon metabolism, several key enzymes are missing. 

Shewanella strains also have only one set of phosphotransferase system 

to uptake simple sugar like glucose through the membrane. So, it is unclear 

that Shewanella strains can utilize six-carbon carbohydrates (e.g. glucose) 

as carbon and energy source. In this study, Shewanella oneidensis MR-1 

was metabolically engineered to have the ability of glucose uptake and 

utilization as carbon and energy source. Glucokinase (glk) and 

glucose-facilitated diffusion protein (glf) of Zymomonas mobilis were 

cloned into a broad host range plasmid and introduced to S. oneidensis 

MR-1 to help glucose uptake and utilization. This recombinant Shewanella 

strain was used for single cell based MFC system in order to confirm 

the glucose utilization and concomitant with current production. The 

results indicated that cloned genes (glf and glk) were functional for glucose 

utilization in S. oneidensis MR-1 wild type. We also found that this 

modification was effective to two Shewanella strains, S. putrefaciens 

CN-32 and S. loihica PV-4. 

Keywords: Shewanella, Glucose, Electricity 

Investigation of Candidate Strains for Application of Microbial 

Electrosynthesis 

Saebom LEE 1 , Dong Geon CHOI 1 , In Geol CHOI 2 and In Seop CHANG* 1 

1 


and Technology (GIST), 261 Cheomdan-gwagiro, Buk-gu, Gwangju 500-712, 

Korea. 2 J-11 

College of Life Sciences and Biotechnology, Korea University, 

Anam-dong, Sungbuk-ku, Seoul 136-701, Korea. 


Reducing power which is obtained from microbial metabolism is used 

to produce several valuable chemicals such as ethanol, butanol, and acetate. 

Microbial electrosynthesis is a system that produces chemicals using 

electron. In microbial electrosynthesis, electrons supplied from electrode 

are accumulated to inner cell and are used to produce the reducing power. 

The accumulated reducing power can be used to produce chemicals. 

Geobacter and Shewanella strains, which are well known as metal reducing 

bacteria, can deliver electrons to outer metal ions through the arrangement 

of cytochrome C protein in the cell membrane. Recently, as reverse 

electron flow of Shewanella strain was revealed, it was known to move 

outer electrons to inside through cytochrome C protein. Based on this 

point, we selected candidate strains which have cytochrome C protein 

in the cell membrane and can produce valuable chemicals though microbial 

reaction. We finally selected Carboxydothermus hydrogenoformans strain 

Z-2901 and Dethiobacter alkaliphilus AHT 1 which have cytochrome C 

protein in extracellular and are able to produce valuable chemicals. Two 

candidate strains will be used for microbial electrosynthesis. 

Keywords: Microbial electrosynthesis; Reverse electron flow

Production of Bio-gas From Moon Jellyfish (Aurelia aurita) 

Ji-Youn KIM 2 and Jae-Hwa LEE* 1 

J-12 

1 

College of Medical and Life Science, Silla University, Busan 617-736, Korea. 

2 

Medical and Life Science, Silla University, Busan 617-736, Korea. 

*Corresponding author: kjy6504@hanmail.net 

The recent study of a jellyfish has caused a hindrance to sea fishery. 

Presently, jellyfish is thrown away through a separator system in the sea. 

The objective of this work was to produce bio-gas from Aurelia aurita 

using anaerobic digestion. The bio-gas includes the hydrogen or the 

methane gas. It relates that jellyfish is effectually changed into the 

renewable energy. In previous study, optimal culture conditions of jellyfish 

have been established, and based on these, and a research including the 

jellyfish amount of addition, the method of treatment, the pH control. 

It optimized with the jellyfish amount of addition with the amount of 

3 g. And the effect of substrate heat treatment, sonication treatment and 

freeze treatment conditions on hydrogen and methane gases production 

in batch experiments. The optimal treatment condition was found to be 

Freeze treated at -70℃ for 20 min. Laboratory-scale experiments were 

carried out in completely mixed bioreactors, 5 L capacity with 4 L worked. 

Optimal amount of cultivation for organics in the step of pretreatment 

was 120 mL Aurelia aurita and used 40 mL wastewater sludge. 

Keywords: Bio-gas, Jellyfish (Aurelia aurita), Anaerobic digestion 

Ag Coated Hydroxyapatite [+Polyacrylonitrile(nanofiber), 

+Polyurethane(film)] Inhibit Growth of Microorganisms 

Kwang-Hwan JHEE* 1 , Jung-Hyun KIM 1 , Byung-Gil MIN 2 and 

Ju-Yeon KIM 2 

1 

Dept. of Applied Chemistry, Kumoh National Institute of Technology, Gumi 

730-701, Korea. 2 J-13 

Dept. of Nano-Bio Textile Engineering, Kumoh National 

Institute of Technology, Gumi 730-701, Korea. 

*Corresponding author: khjhee@kumoh.ac.kr 

Hydroxyapatite (HAp) is a calcium phosphate ceramic that is used as 

a biomaterial. It is chemically similar to the mineral component of bones 

and hard tissues in mammals and its ability to form a strong bond to 

the human hard tissue. Silver (Ag) and Ag-containing compounds have 

been exhibited strong antimicrobial activity. Ag-based antimicrobials draw 

much attention because of the low toxicity of the active Ag ion to human 

cells. We studied the antibacterial effects of Ag containing nanofibers 

(HAp/Polyacrylonitrile, PAN) and films (HAp/Polyurethane, PU). PAN, 

PU and HAp were used for the preparation of electrospinning nanofibers 

and films. Ag containing nanofibers and films were prepared by 

electrospinned with Ag [2.5-5% in DMF(fiber) or H2O/MEK(film)] 

together or soaked into Ag containing DMF(fiber) or H2O/MEK(film) 

solution (1,000 ppm). The antimicrobial effects were investigated by paper 

disc or shaking flask method against E. coli, K. pneumoniae (Gramnegative) 

and S. aureus (Gram-positive). Our data exhibited high 

antibacterial activity against gram-negative and gram-positive both. 

Therefore Hap coated Ag containing fibers and films might be a good 

candidate for the medicinal textile like bandage and baby clothes using 

their antimicrobial activity. 

Keywords: Hydroxyapatite, Antimicrobial activity, Silver 


Oriental Medicines Showing Antifungal Activity Against 

Raffaelea quercus-mongolicae 

Yongsub YI* 1,2 and Jaejoung KIM 2 

J-14 

1 

Department of Herbal Cosmetics Science, Hoseo University, Asan, Korea. 

2 

Department of Biochemistry, Hoseo University, Asan, Korea. 

*Corresponding author: yongsub@hoseo.edu 

Oriental medicines showing antifungal activity against tree wilt disease 

pathogen, Raffaelea quercus-mongolicae, were studied. 16 oriental 

medicines were extracted and assayed antifungal activity against the 

pathogen by disk diffusion assay. Coptidis Rhizoma showed high 

antifungal activity against Raffaelea quercus-mongolicae. Extract of 

Coptidis Rhizoma was fractioned by LC chromatography. Each fractions 

was assayed antifungal activity by disk diffusion assay. More research 

was studied by HPLC and MASS analysis. 

Keywords: antifungal activity, Raffaelea quercus-mongolicae 

Eco-Monitoring of Algal Blooming in Sooyoung Bay 

Hui-Chih WANG 2 , Cora HANESCH 1 , Annalena PEDREIRO 1 , 

Nadine ROYLA 1 , Jessica SCHULZE 1 , Binod PRASAD 1 , Byung-Gook LEE 3 

and Man-Gi CHO* 1 

1 

Department of Bio-Chemical Engineering, Dongseo University Graduate School, 

Busan 617-716, Republic of Korea. 2 Department of Bioprocess Engineering, 

Friedrich-Alexander-University of Erlangen Nürnberg, Busan Campus 618-230, 

Republic of Korea. 3 J-15 

Institue of Ambient Intelligence, Dongseo University 

Graduate School, Busan 617-716, Republic of Korea. 

*Corresponding author: mgcho@gdsu.dongseo.ac.kr 

The purpose of this work is to diagnose the algal blooming based on 

Global Positioning System (GPS) and the fluorescence measurement using 

ToxY-Pam in the Sooyoung Bay. Especially there the samples were taken 

because of the observed algal blooming. The used device measured the 

minimum fluorescence (F0) and the maximum quantum yield of 

photosystem II (Fv/Fm) from the taken samples.The samples were taken 

in December and March. The Chlorophyll Fluorescence values of the 

samples collected in December were lower as compared to March. 

Therefore the collected data can be used for forecasting the onset of algal 

blooming. 

Keywords: Algal Blooming, Fluorescence, Eco-Monitoring 


399

Thermodynamic and Kinetic Analyses of the Binding of a Bacterial 

Expansin (BsEXLX1) to Plant Cell Wall Components 

Injung KIM 2 , Hyeok-Jin KO 2 , In-Geol CHOI 2 , Douglas S. CLARK 3 , 

Tae-Wan KIM 3 and Kyoung Heon KIM* 1 

1 

School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, 

Korea. 2 Biotechnology, Korea University, Seoul 136-713, Korea. 3 J-16 

Energy 

Biosciences Institute, Department of Chemical &Biomolecular Engineering, 

University of California, Berkeley, CA, 94720, USA. 

*Corresponding author: khekim@korea.ac.kr 

BsEXLX1 is a protein belonging to expansin family, found in a soil 

bacterium, Bacillus subtilis, and was previously shown to possess the 

synergistic activity with cellulase on the cellulose hydrolysis. In this study, 

binding characteristics of BsEXLX1 to plant cell wall components were 

thermodynamically and kinetically investigated. At 4°C, the values of 

binding constants, such as Ka and [No], obtained from Langmuir isotherm 

were 6.73´10 6 M -1 and 54 nmol/g cellulose, respectively. Also, the binding 

of BsEXLX1 to cellulose was found to be an endothermic but spontaneous 

reaction (∆Gº=-36.2 kJmol -1 and ∆Hº=57 kJmol -1 ). Generally, the 

BsEXLX1 exhibited an enhanced binding efficiency under acidic 

conditions. Also, the amount of bound protein increased in the presence 

of NaCl. Against cellopentaose and cellohexaose, the binding activity of 

BsEXLX1 was negligible in the isothermal titration calorimetry. Against 

the pretreated lignocellulosic biomass with different compositions of 

cellulose, hemicelluloses and lignin, the preferential binding of BsEXLX1 

to lignin was observed. This finding could be applicable in cellulose-based 

pulp and textile industries or biofuel production for using the bacterial 

expansin as a non-blocking agent to enhance the accessibility of cellulose 

to enzymes. 

Keywords: Bacterial expansin, Cellulose, Lignocellulose 

Screening of an Esterase/lipase-Producing Bacteria from 

Gastropods 

Jung-Hee WOO* 1 , Sun Mee HONG 1 , Jun-Tae KIM 1 , Tae-Hyung KWON 1 , 

Hyun-Soo RHO 2 , Jong-Shik KIM 1 , Nyun-Ho PARK 1 and 

Choong-Gon KIM 1 

1 

1Gyeongbuk Institute for Marine Bio-Industry (GIMB), Uljin 767-813, 

Gyeongbuk, Korea. 2 J-17 

2East Sea Research Institute, Korea Ocean Research 

&Development Institute (KORDI), Uljin 767-813, Gyeongbuk, Korea. 

*Corresponding author: jhwoo@gimb.or.kr 

Esterases and lipases are enzymes that catalyze the hydrolysis of 

carboxyl esters and are found in various organisms including animals, 

plants, fungi and bacteria. These enzymes have increased the 

biotechnological interest for a number of industrially significant 

biotransformations, either as hydrolytic or as synthetic catalysts. 

Previously we reported that bacteria isolated 127 strains from six 

gastropods F. oregonensis, N. cumingi, N. constricta, C. fulmen, V. 

emphaticus, JG in June to December 2010 were studied to find out the 

bacterial assemblages associated with local gastropod collected in Uljin, 

the East Sea, Korea. Esterase/lipase-producing bacteria were isolated from 

gastropods. To screen for lipolytic activity, various agar medium 

supplemented with 1% tributyrin (TBN) and tricaprylin (TCN) were used. 

We found that 56 isolates showed lipolytic activity on TBN agar plate 

and 22 isolates showed lipolytic activity on TCN agar plate 

(esterase-producing strains= 34, lipase-producing strains= 22). This study 

presents a first example which discovered esterases/lipases from 

gastropods successfully. [This work was supported by the Marine and 

Extreme Genome Research Center Program, Ministry of Land, Transport 

and Maritime Affairs, Republic of Korea.] 


J-18 

Survival of Escherichia coli O157:H7 and Listeria in Manure 

Compost in Korea 

Sook Young JANG, Sunggi HEU, Eunjung ROH, Dong Hwan LEE, 

Yang Kyun KIM, Mukesh KUMAR, Shweta MALHOTRA, Jong Chul YUN 

and Kyu Seok JUNG* 

Microbial Safety Division, National Academy of Agricultural Science, RDA, 

Suwon, 441-707, Korea. 

*Corresponding author: win258@korea.kr 

On-farm contaminations through contaminated manure were considered 

likely sources of the pathogen for several outbreak. Of particular concern 

are bacteria causing human food-borne illness such as Salmonella species 

and E. coli O157:H7. The objective of this study was to quantify the 

level of environmental impact of temperature on E. coli O157:H7 and 

Listeria survival in manure compost. Commercial manure compost 

(manure 60%, sawdust 40%) was inoculated with E. coli O157:H7 and 

Listeria. Composts were incubated at four different temperature (10, 25, 

35, and 55℃) and consistent moisture content (55%). Samples had been 

collected during 49 days depending on the given conditions. E. coli 

O157:H7 and Listeria persisted for up to 1 day in compost at 55℃ and 

49 days at three different temperature (10, 25 and 35℃). It is noted that 

as the temperature increased, the number of E. coli O157:H7 and Listeria 

gradually decreased over time. The results indicate that E. coli O157:H7 

and Listeria persisted longer under low temperature condition. This study 

will provide useful and practical guidelines to applicators of soil in 

deciding appropriate handling and time frames for land application for 

sustainable agriculture. 

Keywords: E. coli O157:H7, Listeria, survival 

Microbiological Safety of Gamma-Irradiated Porcine Skin for 

the Development of Dressing Material 

Jong-Il CHOI 1 , Eu-Ri JO 1 , Pil-Mun JUNG 1 , Jae-Hun KIM 1 , 

Beom-Suk SONG 1 , Jae-Kyung KIM 1 , Jong-Heum PARK 1 , Tae-Woon KIM 2 

and Ju-Woon LEE* 1 

J-19 

1 

Team for Radiation Food Science and Biotechnology,Advanced Radiation 

Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185. 

2 

Rion-Biotechnology Co.,wanju-gun 565-902, Korea. 

*Corresponding author: sjwlee@kaeri.re.kr 

In this study, sensitivities of bacteria and virus in porcine skin by gamma 

irradiation were evaluated to develope the optimum sterilization condition 

for the development of dressing material. Escherichiacoli (E.coli) and 

Bacillus substilis (B. substilis) were used as model pathogens and 

inoculated 106-107log CFU/g. As model virus from porcine, porcine 

parvovirus (PPV), bovine viral diarrhoea virus (BVDV) and poliovirus 

were inoculated 105-106TCID50/g into porcine skin. The D10 value of 

E.coli was determined as 0.25±0.1 kGy. B. substilis showed lower 

radiation sensitivity as D10 was 3.88±0.3 kGy. The D10 values of PPV, 

BVDV and poliovirus were determined as 1.73±0.2, 3.81±0.2 and 6.88±0.3 

kGy, respectively. These results suggest that gamma irradiation can reduce 

the risk of contamination and offer the basic information to inactivate 

the pathogens as microbiologically safe dressing material of porcine skin. 

Keywords: radiation sensitivity, bacteria and virus, porcine skin

Optimal Culture Conditions of Microalgae Haematococcus 

pluvialis 

Hyun-Jin PARK 1 , Young-Hwa KIM 2 and Jae-Hwa LEE* 1 

1 

Department of Bioscience and Biotechnology, College of Medical and Life 

Science, Silla University, Busan 617-736, Korea. 2 J-20 

Department of Pharmaceutical 

Engineering, College of Medical and Life Science, Silla University, Busan 

617-736, Korea. 

*Corresponding author: phj0386@naver.com 

The green microalgae Haematococcus pluvialis (H. pluvialis) produces 

large amounts of astaxanthin. The morphological change of H. pluvialis 

from green vegetative cells to red resting cyst cells is accompanied by 

an enhancement in astaxanthin production. To find out high mass culture 

conditions of H. pluvialis, some important growth factors, such as 

temperature, initial pH, culture media, CO2, and air effect were tested. 

The optimal growth condition was found to be as follows: temperature 

35℃, initial pH 6.5, 4X MBBM medium. Maximum growth rate of H. 

pluvialis was obtained at a 5% CO2(0.81 g/L), air(1.06 g/L) environment. 

From these conditions, it could be possible to cultivate H. pluvialis for 

large-scale high density. 

Keywords: optimal culture, Haematococcus pluvialis, marine microalga 

Heterologous Expression of Metagenome-Derived Wax Ester 

Synthase in Escherichia coli and Arabidopsis thaliana. 

Nam-hee KIM 1 , Ji Hye PARK 1 , Eun Sook CHUNG 2 , Jin Cheol KIM 3 , 

Myung Hwan LEE 1 , Weixin TAO 2 , Eul Chul HWANG 1 , Jai Heon LEE 2 and 

Seon Woo LEE* 1,2 

1 

Department of Applied Biology, Dong- A University, Busan 604-714, Republic 

of Korea. 2 Department of Medical Bioscience, Dong- A University, Busan 

604-714, Republic of Korea. 3 J-21 

Chemical Biotechnology Research Center, KRICT, 

Deajeon 305-600, Republic of Korea. 

*Corresponding author: seonlee@dau.ac.kr 

We have previously selected a metagenome clone carrying an ORF 

encoding a wax ester synthase (WES) like protein. Deduced amino acid 

sequences of the ORF exhibited 30% and 22% identities to WES from 

Acinetobacter baylyi and Arabidopsis thaliana, respectively. The 

WES-like protein was functional as a wax ester synthase, when wes gene 

was expressed in Escherichia coli fed by fatty alcohol, 10 mM of 

hexadecanol, as a precursor for wax ester synthesis. The unique wax esters 

produced in E. coli carrying wes gene were identified as wax esters of 

various chain lengths by GC-MS analysis. The wes gene was also 

introduced into a model plant, Arabidopsis thaliana, to produce wax esters 

in a plant system. Transgenic plants carrying the wes gene were obtained 

by Agrobacterium-mediated transformation. Expression of wes gene in 

the transgenic plants was confirmed by RT-PCR. Production of wax esters 

and other phenotypic changes in the transgenic plants are under 

investigation. 


J-22 

Community Structure Analysis of Cyanobacteria using 16S rRNA 

and Phycocyanin Gene in Korean Reservoirs 

Kyoung-Hee OH, Seung-Hee SHIN, Eun-Hye PARK, Mi-Na YU and 

Young-Cheol CHO* 

Dept. of Environmental Engineering, Chungbuk National University, Cheongju 

361-763, Korea. 

*Corresponding author: choy@cbnu.ac.kr 

The community structure of cyanobacteria was examined with 16S 

rRNA and phycocyanin intergenic spacer (PC-IGS) genes as target. 

Although DGGE analysis with 16S rRNA gene reflected the changes of 

cyanobacterial community structure with fluctuations of environmental 

factors, the resolution was restricted to the genus level because of its 

homogeneity. On the other hand, analysis of PC-IGS gene sequences 

showed that this gene can classify cyanobacteria in strain level, indicating 

it is considered as an adequate target gene to analyze the community 

structure of cyanobacteria in the environmental samples. Moreover, the 

PC-IGS gene sequences of Microcystis strains in the investigated reservoirs 

were divided into two clades and each clade exclusively included toxicor 

non-toxic Microcystis, proposing PC-IGS gene can be used to monitor 

toxigenic Microcystis at the cyanobacterial bloom events. [Supported by 

grants from National Science Foundation of Korea] 

Keywords: Cyanobacteria, PC-IGS, Community structure 

J-23 

Ethanol Fermentation using Main Sugar in Laminaria japonica 

Geun Hyub KIM, Sung-Mok LEE and Jae-Hwa LEE* 

Department of Bioscience and Biotechnology, College of Medical and Life 

Science, Silla University, Busan 617-736, Korea. 

*Corresponding author: jhalee@silla.ac.kr 

The excessive use of fossil fuels has recently been causing many 

problems around the world such as global warming. Therefore, alternative 

energy source has been of interest, much research as been performed in 

order to develop alternative energy. This study examined the production 

of bio-ethanol from the brown-algae main sugar compound. The result 

of ethanol fermentation used four-bacteria, ethanol production of 

Zymomonas mobilis KCTC 1534 with different carbon sources (alginate, 

laminaran, mannitol and Laminaria japonica) confirmed 2.31 g/L, 1.32 

g/L and 2.36 g/L respectively. The maximum cell growth with laminaran 

reached 1.42 (O.D. 600 nm), but other substrate was not significant 

increases in the fermentation period. The maximum cell growth of 

Zoogloea ramigera KCCM 40423 was reached approximately 2.61 and 

2.37 (O.D. 600 nm) with mannitol and laminaran, respectively. And then 

ethanol production was 0.97 and 0.236 g/L. Therefore, Laminaran like 

to be limitations to what ethanol production, but can consume for cell 

growth increase. Streptomyces venezuelae KCCM 40310, Paracoccous 

denitrificans KCCM 40267 was not detected ethanol production, but cell 

growth was significant increases. 

Keywords: ethanol fermantation, Laminaria japonica, polysaccharides 


401

Identification of Novel Sugar Pathway Genes and their 

Organization from an Insect Gut Metagenome 

Chang-Muk LEE* 1 , Sang-Hong YOON 1 , Soo-Jin KIM 2 , So-Hyeon SEO 1 , 

Young-Seok LEE 1 , Jung-Keun PARK 1 , Han-Chul KANG 1 and 

Bon-Sung KOO 1 

J-24 

1 

Functional Biomaterial Division, National Academy of Agricultural Science, 

2 

Rural Development Administration, Suwon 441-707, Korea. National 

Agrobiodiversity Center, Academy of Agricultural Science, Rural Development 

Administration, Suwon 441-707, Korea. 

*Corresponding author: changmuk@rda.go.kr 

Insect gut symbiotic microbiota play essential roles in the recycling 

various organic materials, and may thus be exploited as a simple system 

for processing daily food wastes. To detect the contributing microbial 

genes, we have constructed 92,544 metagenomic fosmid clones using the 

larval intestinal microbiome from black soldier fly, Hermetia illucens. 

The hydrolytic enzymes encoded by uncultured microorganisms were 

subjected to substrate hydrolysis analysis. Initial screening of the libraries 

revealed a clone that uses carboxymethyl cellulose as a sole carbon source. 

Functional screening and metagenome walking with shot-gun libraries 

revealed two novel gene clusters conferring multiple sugar pathways. The 

cluster was 36kb in size and transcribed as polycistronic units. They 

include glycosidase-related sugar pathway genes that are arranged as sugar 

transferase genes, glycan processing genes, transporter genes, and putative 

transcription factors. The presence of insertion sequences between clusters 

suggests that the metagenome-derived clone has acquired the specific 

organization by lateral gene transfer [Supported by grants from the Rural 

Development Administration project number PJ0067032011]. 

Keywords: Metagenome, Hermetia illucens gut, Glycosidase 

Development of Transgenic Daphnia magna Expressing Enhanced 

green Fluorescent Protein 

Thai Hoang LE 1 , Sung-Kyu LEE 2 , Yang-Hoon KIM 3 and Jiho MIN* 1 

1 

Department of Bioprocess Engineering, Chonbuk National University, 664-14 

Duckjin-dong, Jeonju 561-756, South Korea. 2 Toxicology Research Center, Korea 

Research Institute of Chemical Technology, Daejon, 305-600, Korea. 3 J-25 

School of 

Life Science, Chungbuk National University 410 Sungbong-Ro, Heungduk-Gu, 

Cheongju 361-763, South Korea. 

*Corresponding author: jihomin@jbnu.ac.kr 

To establish a novel system for a rapid detection of environmental stress, 

a transgenic Daphnia magna able to express a green fluorescent protein 

has been created. A 32bp DNA fragment containing the 18s-ribosome 

constitute promoter of daphnia was generated by in-vitro annealing the 

commercial primers at 80 o C. This target DNA was inserted ahead to the 

gfp gene in the EGFP vector (Enhanced Green Flourescent Protein). The 

successful recombinant plasmid confirmed by the DNA sequencing was 

introduced into the earlier developing-state daphnia eggs (round shape) 

using microinjection technique. After 48h incubation in the 20 o C chamber, 

the injected eggs able to develop to juveniles were observed at the 488nm 

wavelength in the confocal fluorescent microscope to confirm the presence 

of green fluorescent protein. Our results show that the green fluorescent 

protein was substantially found in the upper part of the transgenic 

organisms (e.g., head, back) while no fluorescent signal was detected in 

the control organism. It demonstrated that a transgenic Daphnia magna 

expressing green fluorescent protein was firstly generated and would be 

promising to build a novel testing system for easily monitoring 

environmental stress. 

Keywords: Daphnia magna, transgenic, fluorescent 


Isolation and Characteristics of Antagonic Microorganisms as 

Nematicide Against Root-knot Nematode of Fruit Vegetables 

(Oriental Melon and Cucumber) 

Beam Soo KIM 1 , Keun-Hyung LEE 1 , Young-Won KANG 1 , Chang-Su KIM 1 , 

Se-Ho MYEONG 1 , Joung-Ja KWON 2 , Chung-Sig CHOI 3 and 

Gi-Seok KWON* 1 

1 

School of Bioresource Sciences, Andong National University, Andong 760-749, 

Korea. 2 Cheongsong Agricultural Technology Center Cheong Song Gun, 

CheongSong 763-802, Korea. 3 J-26 

Bio Industry Institute, HansBio Co.,Ltd. Andong 

760-380, Korea. 

*Corresponding author: suyaa18@hanmail.net 

This study was conducted to screen the antagonic microorganisms on 

root-knot nematode and to utilize in its control. The effect of nematocidal 

activity of antagonic microorganisms were tested on the nematicide of 

root-knot nematode, Showing nematocidal activities of 88 strains isolated 

from various fruit Vegetables field and soil. In vitro study, all tested 

treatments had nematicidal effect on nematode juveniles 24 and 48 hours 

from exposures in the 24-well plate. Among them, BS-17, BS-51(2), 

BS-51(3), BS-52(2), and BS-84(2) were isolated. The highest percentage 

of nematode mortality was achieved by application of BS-17(96.38%), 

BS-51(2)(94.11%), BS-51(3)(95.38%), BS-52(2)(94.59%), BS-84(2) 

(98.48%). Under greenhouse conditions, isolated antagonic microorganisms 

treatments were effective in reducing population numbers of root-knot 

nematode and root gall formation in compared to non-treatments. 

Keywords: Root-knot nematode, Meloidogyne. Antagonic microorganisms, 

Nematocidal activity, Ne 

Effect of Plant Growth-Promoting Rhizobacteria on Growth of 

Chinese cabbage Plant under Salt Stress 

Hee Sun KOOK 1 , Gun-Woon LEE 1 , Song-Joong YUN 2 , Jong-Chan CHAE 1 

and Kui-Jae LEE* 1 

J-27 

1 

Division of Biotechnology, Chonbuk national University, Iksan 570-752, korea. 

2 

Division of Agriculture Life sciences, Chonbuk National University, Chonju 

561-765, Korea. 

*Corresponding author: k722954@naver.com 

To gain understanding of responses to salinity stress by rhizobacteria 

at physiological level, we isolated 15 strains of plant growth-promoting 

bacteria from soil samples taken in the reclaimed tidelands. These 15 

strains have been identified as Pseudomonas, Herbaspirillum, Enterobacter, 

Serratia, and Stenotrophomonas by 16S rRNA gene sequence analysis. 

With the exception of Serratia sp., all strains can product indole acetic 

acid (IAA) and IBA (butyric acid). Among the selected 15 strains, eight 

strains only showed 1-aminocyclopropane-1-carboxylate (ACC) 

deaminase activity and rhizobacterial ACC deaminases were detected by 

polyclonal anti-ACCD antibody. To evaluate whether rhizobacteria 

inhabiting saline environments confer the resistance against salt stress to 

Chinese cabbage plant, cabbage seeds were sown in soil inoculated with 

respective bacterial strains. And then, the early seedling growth was 

examined under 0 or 100 mM NaCl solution treatment for 2 weeks after 

germination. Results of our study exhibited that inoculation with selected 

strains increased the shoot length ranging from 107 to 141%, compared 

to that of the uninoculated control. These results indicate that the 

rhizobacterial stains had the potential to promote the growth of cabbage 

plant under saline condition. 

Keywords: plant growyh-promoting rhizobacteria (PGPR), Chinese cabbage, 

NaCl, Salt stress

J-28 

Characterization of Microbial Community of the Ammonia 

Oxidizing Enrichment Culture from Wastewater Treatment Plants 

Hyoun Duk JUNG and Eun Young LEE* 

Department of Environmental Engineerin and Environmental Microbiology, 

Suwon University, Gyeonggi 445-743, Korea. 

*Corresponding author: ley@suwon.ac.kr 

Ammonia oxidizing enrichment culture carried from S wastewater 

treatment plant was performed in the AOB medium. The microbial 

community in wastewater treatment plants were characterized using 

potential carbon availability method. For the Ecoplate (Biolog, USA) 

analysis, those samples were inoculated and cultured at 20℃. Color 

changes of individual well in the Ecoplate according to cell growth were 

measured at 595nm and average well color development (AWCD) was 

calculated. The order of AWCD values of S1(enrichment culture), 

S2(aeration tank), S3(effluent)and S4(influent) were 0.36,1.18,1.12 and 

0.98, respectively.The Shannon indies were 2.31(S1), 3.35 (S2),3.37(S3) 

and 3.38(S4),respectively. In addition, Bacterial community was 

characterized using 16S rDNA PCR and denaturing gradient gel 

electrophoresis (DGGE) fingerprinting method. the similarities of DGGE 

fingerprints were 7% between S1 and S2, 11% between S1 and S3, 7% 

between S1 and S4, 11% between S2 and S3, 8% between S2 and S4, 

13% between S3 and S4 of the ammonia oxidizing enrichment culture 

from wastewater treatment plants. 

Keywords: DGGE, Ecoplate, wastewater 

Reduction of Tellurite and Formation of Tellurium Nanostructures 

by Shewanella oneidensis MR-1 

Dong-Hun KIM 1 and Hor-Gil HUR* 1,2 

J-29 

1 2 

School of Environmental Science and Engineering. International Environmental 

Analysis and Educational Center, Gwangju Institute of Science and Technology, 

Gwangju 500-712, Republic of Korea. 

*Corresponding author: donghun@gist.ac.kr 

The metalloid oxyanion tellurite(TeO3 2- ) is highly toxic to most 

organisms and specific mechanisms explaining its toxicity are not well 

known to date. In this study we determine the reduction of tellurite and 

formation of tellurium by S. oneidensis MR-1. Strain MR-1 can reduce 

tellurite [TeO3 2- ] to elemental tellurium [Te(0)] under anaerobic conditions. 

This reduction result in the formation of unique crystalline Te 

nanoarchitectures as end products. The localization of needle-like shaped 

Te nanostructures are controlled by culture conditions. EDX and SAED 

indicate that nanostructures are composed entirely of Te and are crystalline. 

Furthermore, c-type cytochrome deletion mutants and random mutants 

from strain MR-1 shown lower reduction rate of tellurite than wild type 

and different morphologies on tellurite contained solid medium. Additional 

works needs for describe more details about molecular mechanisms of 

the tellurite reduction and formation of nanostructures. This research offers 

the potential to combine remediation of contaminants with the development 

of environmental friendly manufacturing pathways for novel biogenic 

nanominerals and recovery of rare metals from industrial waste water. 

Keywords: Keywords: Shewanella, Tellurium, dissimilatory metal-reducing 

bacteria (DMRB), nano structure 


J-30 

Improved Performance of a Microbial Electrolysis cell using a 

Membrane Electrode Assembly Cathode 

Yuhong JIA, Jae Hun RYU, Cho Hui KIM, Woo Kyung LEE, 

Thi Van Trinh TRAN, Hyo Lee LEE and Dae Hee AHN* 

Department of Environmental Engineering and Biotechnology, Myongji 

University, Yongin 449-728, Korea. 

*Corresponding author: dhahn@mju.ac.kr 

Microbial electrolysis cell is a device which can produce hydrogen gas 

from biomass through microbial catalyzed process and thus reduce the 

organic matter. For the real application in wastewater treatment, the 

scale-up of microbial electrolysis cell is an important issue but few tests 

were conducted with relatively large size. In this study, a 3.7 L microbial 

electrolysis cell equipped with a membrane electrode assembly cathode 

was designed and tested. The internal resistance was examined, hydrogen 

generation and organic removal performance was investigated under 

different conditions. An internal resistance of 7 Ω was examined from 

the reactor with a relatively large working volume. The MEC achieved 

a coulombic and cathodic efficiency of 55 % and 74 % respectively at 

an applied voltage of 1.2 V, results a maximum overall hydrogen efficiency 

of 41 %. This corresponding to a volumetric hydrogen production rate 

of 0.12 L-H2/LA-day (A = Anodic liquid volume) at a hydrogen yield 

of 1.64 mol H2/mol acetate and a current density of 2.8 A/m 2 . The 

hydrogen performance at voltages below 0.8 V was limited, which might 

be due to the high potential losses in large system. The results obtained 

in this study could help to further develop pilot-MEC for practical 

applications. 

Keywords: Microbial electrolysis cell, Hydrogen production, Membrane 

electrode assembly 

Anti-bacteria and Anti-fungal Effects of Herbal Oil made from 

Halvateria acuminata. 

Soo Ji KIM* 1 , L.M Udaya CABRAL 2 , Jin Young HONG 1 , Chang Wook 

JO 1 , Mi Hwa JUNG 1 , Young Hee KIM 1 and Jung Eun CHOI 1 

1 

Conservation Science Division, National Research Institute of Hultural Heritage, 

82 Munji-ro, Yuseong-gu, Daejeon, 305-380, Korea. 2 J-31 

Conservation and 

Preservation Division, National Library and Documentation Services Board, 

No.14, Independence Avenue, Colombo-7, Sri Lanka. 

*Corresponding author: feeelsj@naver.com 

An Ola leaf manuscript, which consists of words carved on an Ola 

leaf and filled up with mixture of herbal oil made from Halvateria 

acuminata and charcoal, is one of the native writing media in Sri Lanka. 

According to historical records, Ola leaf manuscripts had been used from 

1 st century A.D to 18 th century A.D. From recent findings that Ola leaf 

manuscripts have been preserved well for the past 400years and are in 

good state of preservation, it is supposed that this oil preserves Ola leaves 

against environment factors and biological factors such as fungi and 

insects. To evaluate the anti-biological susceptibility of the herbal oil, 

the molds isolated from wooden printing blocks in Janggyeong Panjeon 

of Haeinsa temple in Korea and bacteria isolated from Ola leaf are cultured. 

After spreading of microorganisms suspension on agar plate, a disk paper 

containing certain volume of herbal oil is put on the agar plate. From 

the experiment, it is found that the herbal oil exhibits a clear zone, which 

is optically clear and inhibits growth of microorganisms, against some 

molds and bacteria. The result indicates that herbal oil from the plant 

Halvateria acuminata has the anti-bacteria and anti-fungal properties. 

Keywords: herbal oil, anti-bacteria, anti-fungal 


403

J-32 

Enhancement of Biohydrogen Production in [NiFe]-hydrogenase 

Expressing Recombinant Escherichia coli by Introducing Light 

Acceptor 

Younghwa JO, Jaoon Y.H. KIM, Byung Hoon JO and Hyung Joon CHA* 

Pohang University of Science and Technology, Pohang 790-784, Korea. 

*Corresponding author: hjcha@postech.ac.kr 

Biohydrogen production can be one of solutions for energy crisis. 

Previously, we successfully demonstrated the production of biohydrogen 

by constructing [NiFe]-hydrogenase-expressing recombinant E. coli. 

Although [NiFe]-hydrogenase showed relatively high oxygen-tolerance 

compare to other hydrogenases, still biohydrogen production efficiency 

was low. In this research, we introduced proteorhodopsin, light acceptor 

for prokaryote working as proton pump, into [NiFe]-hydrogenaseexpressing 

recombinant E. coli for utilizing light energy for producing 

biohydrogen. Proteorhodopsin need a chemical named retinal for working 

as proton pump by using light energy but E. coli does not have a pathway 

to synthesize it. In order to give E. coli an ability to synthesize retinal, 

we expressed five foreign genes expressing enzymes for retinal synthesis 

pathway. By introducing light accepting system into E. coli expressing 

[NiFe]-hydrogenase, we could get enhanced biohydrogen productivity. 

Keywords: Biohydrogen, [NiFe]-hydrogenase, proteorhodopsin 

Novel Probiotic Strains for American Foulbrood and Nosemosis 

of Honeybee 

Yoon Jeong JANG 1 , Byoung Joo SEO 1 , Irfan RATHER 1 , Uk Han CHOI 1 , 

Jeonghuei LIM 1 , Kook Hee KANG 2 , Byoung Su YOON 3 and 

Yong-Ha PARK* 1 

J-33 

1 

Department of Applied Microbiology and Biotechnology, Yeungnam University. 

2 3 

Korea Institute of Science and Technology Information, Daejeon. Department 

of Life Science, Kyonggi University, Suwon, Kyonggi-Do. 

*Corresponding author: joos02@ynu.ac.kr 

American Foulbrood of honeybees (Apis mellifera), caused by the 

Gram-positive bacterium Paenibacillus larvae is one of the most serious 

diseases affecting the larval and pupal stages of honeybees. Presently, 

Nosema ceranae and Nosema apis seems to be the main microsporidian 

infection in honeybees. The aim of this study was to evaluate lactic acid 

bacteria strains isolated from Kimchi and to study its inhibitory effects 

against Paenibacillus larvae subsp. larvae KCTC 3744 and Nosema 

trichoplusiae ATCC 30702. In all, 3,500 strains were screened and three 

highly active bacteria were selected and were identified as L. plantarum 

YML001, L. plantarum YML004, Leu. citreum KM20. The optical density 

of P. larvae KCTC 3744 at OD600nm showed drastic decrease in cell 

growth as compared to control. Three groups of honey bees (N=25) were 

given Nosema Spores at a concentration of 10,000 spores per ml with 

addition of 1% of supernatant of three strains. The bees were incubated 

at 37°C and samples were taken seven days post infection, and the amount 

of spores in the mid gut was counted. We found that the three strains 

inhibited the growth of P. larvae KCTC 3744 and N. trichoplusiae ATCC 

30702 and provided new perspective for probiotics of honeybees. 

Keywords: American Foulbrood, Nosemosis, Probiotics 


Analysis of Bacterial Community in Electrochemical Bioreactors 

Cultivated with Carbon Dioxide as Sole Carbon Source 

Doo Hyun PARK 2 , Jinsu KIM 2 , Bo Young JEON 2 and Il Lae JUNG* 1 

1 

Department of Radiation Biology, Environmental Radiation Research Group, 

Korea Atomic Energy Research Institute, Daejeon 305-353, Korea. 2 J-34 

Department 

of Biological Engineering, Seokyeong University, Seoul 136-704, Korea. 


Bacterial community was isolated from different sources that were forest 

soil, aerobic wastewater treatment reactant, anaerobic wastewater digestive 

reactant, farm field soils, mixture of isolated bacterial strains. Bacterial 

communities grown in the different bioreactors were analyzed by 

temperature gradient gel electrophoresis technique. Different bacterial 

communities were enriched and selected during cultivation with the 

electrochemical reducing power as a sole energy source. TGGE pattern 

is not an indicator to show all of the bacterial communities grown in 

the electrochemical bioreactors; however, the bacterial species capable of 

growing with carbon dioxide or surviving in the condition saturated with 

carbon dioxide may be enriched selectively. According to the bacterial 

species identified based on the sequence homology, various bacterial 

species may fix metabolically carbon dioxide. Acknowledgement This 

work was supported by the New &Renewable Energy of the Korea Institute 

of Energy Technology Evaluation and Planning (KETEP) grant funded 

by the Korea governmental Ministry of Knowledge Economy 

(2010T1001100334) 

Keywords: electrochemical reducing power, carbon dioxide fixation, 

Bacterial community diversity 

Screening of Protease-Producing Bacteria from Gastropods 

Jung-Hee WOO* 1 , Sun Mee HONG 1 , Jun-Tae KIM 1 , Jin-Wook LEE 1 , 

Hyun-Soo RHO 2 , Jong-Shik KIM 1 , Nyun-Ho PARK 1 and Choong-Gon KIM 1 

1 

1Gyeongbuk Institute for Marine Bio-Industry (GIMB), Uljin 767-813, 

Gyeongbuk, Korea. 2 J-35 

2East Sea Research Institute, Korea Ocean Research 

&Development Institute (KORDI), Uljin 767-813, Gyeongbuk, Korea. 

*Corresponding author: jhwoo@gimb.or.kr 

Microbial proteases are the most important group of industrial enzymes 

such as food, detergent, pharmaceutical, and waste management industries. 

Previously we reported that bacteria isolated 127 strains from six 

gastropods F. oregonensis, N. cumingi, N. constricta, C. fulmen, V. 

emphaticus, JG in June to December 2010 were studied to find out the 

bacterial assemblages associated with local gastropod collected in Uljin, 

the East Sea, Korea. Protease-producing bacteria were isolated from 

gastropods. To screen for proteolytic activity, various agar medium 

supplemented with 0.5% skim milk were used. We found that 38 isolate 

strains showed proteolytic activity on skim milk agar plate (10 o C = 16 

strains, 25 o C = 22 strains). This study presents a first example which 

discovered microbial proteases from gastropods successfully. [This work 

was supported by the Marine and Extreme Genome Research Center 

Program, Ministry of Land, Transport and Maritime Affairs, Republic of 

Korea] 

Keywords: Protease, Proteolytic activity, Gastropods

J-36 

Simultaneous Quantification of Cyanobacteria and Microcystis 

spp. using Quantitative Realtime PCR 

Kyoung-Hee OH, Seung-Hee SHIN, Mi-Na YU, Eun-Hye PARK and 



361-763, Korea. 


Microcystins are produced mainly by Microcystis sp. at large reservoirs 

in Korea. Therefore, quantification of Microcystis is required to monitor 

the potential for algal-toxin production. It is difficult to count Microcystis, 

however, because these mostly form scum. In this study, new method 

was developed to quantify cyanobacteria and Microcystis simultaneously 

using quantitative realtime PCR. The primers and probe, cyanobacteriaspecific 

and Microcystis-specific primers and Microcystis-specific TaqMan 

probe, were designed from the conserved regions of 16S ribosomal RNA 

gene sequences for cyanobacteria and Microcystis, respectively. It was 

proved as an adequate and reproducible method to quantify cyanobacteria 

and Microcystis in the environmental samples. It is expected that the use 

of this method enhances the accuracy of Microcystis counts and reduces 

the time involved in the analysis of samples by microscopy. Also, this 

method enables us to monitor the formation of cyanobacterial mass 

occurrences in situ and to reveal the factors promoting the growth of 

Microcystis. [Supported by grants from National Science Foundation of 

Korea] 

Keywords: Microcystis, Real-time PCR, Simultaneous quantification 

J-37 

GFP Tagging of p-Cresol Methylhydroxylase in Pseudomonas 

alkylphenolia and Its Dominant Expression in Colonies 

CHO AH RA and Kyoung LEE* 

Dept. of Microbiology, Changwon National University, Kyongnam 641-773, 

Korea. 

*Corresponding author: 0506joara@hanmail.net 

In this study, the chromosome-encoded pcuRCAXB genes that are 

required for p-cresol degradation have been identified by using the newly 

constructed Green fluorescence protein (GFP)-based promoter probe 

transposon in the long-chain alkylphenol degrader, Pseudomonas 

alkylphenolia. The deduced amino acid sequences of the genes showed 

the highest identities at the levels of 65-93% compared to those in the 

databases. The transposon was identified to be inserted in the pcuA gene 

with the promoterless gfp gene being under the control of the pcu catabolic 

gene promoter. The expression of GFP was positively induced by p-cresol 

and was about 10 times higher by cells grown on agar than those in 

liquid culture. This result indicates that P. alkylphenolia additionally 

possesses a protocatechuate ortho cleavage route for p-cresol degradation 

that is dominantly expressed in colonies. 

Keywords: Pseudomonas, transposon 


Isolation and Characteristics of Entomopathogenic Fungi 

Beauveria bassiana as Insectide Agent Against Greenhouse 

Ahitefly(Trialeurodes vaporariorum) 

Chang-Su KIM 2 , Keun-Hyung LEE 3 , Young-Ho NAM 3 , 

Gyeong-Muk KANG 3 , Jin-Won KIM 4 and Gi-Seok KWON* 1 

1 

Departmentl of bioresource Sciences Andong Nation University, Andong 

760-749, Korea. 2 안동대학교. 3 School of bioresource Sciences, Andong Nation 

University, Andong 760-749, Korea. 4 J-38 

Yecheon Agricuture Technology Center, 

Yecheon 757-802, Korea. 

*Corresponding author: ckdtn0629@hanmail.net 

The object of this study is to use Entomopathogenic fungi B.bassiana 

as agent Greenhouse whitefly(Trialeaarodes vaporariorum). Each 4 strains 

were prepared as follows : B.bassiana was parceled by Rural Development 

Administration(J157&J216), Gyougbuk Agricutural Reserch&Extension 

Service(S186) Yecheon Agricultuer Technology Center(B.K). Chitinase, 

protease, lipase are important and involved in the host invasion. In vitro, 

plate assay was used to observe the each enzyme. And chitinase was 

observed by three strains. Also, Protease, lipase was observed by all strains. 

And activity measurement of enzyme is in progress. In vivo, one control 

plot and each four treatment plots was treated to 50 Greenhouse whiteflys 

in a capsicum. After a few days, B.bassiana spore(60g/1L) that was 

mixtuerd the yolk of an egg spraied due volta 500ml in each treatment 

plots and spraied a Neoseiulus californicus in the control plot. Later, 

indicated Greenhouse whitefly that enticed yellow flypaper trap and 

compared control plot with treatment plots. Consequently, B.bassiana was 

a predominant and confirmed as insecticide agent against Greenhouse 

whitefly.[ This study supported by Technology of Development program 

for Agriculture and Foresty, Gyeongbuk 2011] 

Keywords: Beauveria bassiana, Entomopathogenic, Greenhouse whitefly 

J-39 

Effect of Nitrate on the Electricity Generation of Microbial Fuel 

cell 

Jae Kyung JANG*, Jung Eun CHOI, Young Sun RYOU, Sung Hyoun LEE, 

Jong Goo KIM, Youn Koo KANG, Young Hwa KIM and Hyung Mo LEE 

National Academy of Agricutural Science. 

*Corresponding author: jkjang1052@korea.kr 

The microbial fuel cells (MFCs) can convert to useful electricity from 

artificial wastewater including nitrogen source such as nitrate (NO3 - ) and 

also to treat this wastewater. Electrons and protons produced by metabolic 

process of microorganisms in the anode compartment transfer to cathode 

through the out circuit and membrane, respectively. However, when the 

nitrate is included in the wastewater (fuel), it can act the electron donor. 

It will decrease the current value. These studies were conducted to 

investigate the influence of nitrate concentration on the current generation. 

Sodium acetate was used as electron donor and the chemical oxygen 

demand (CODcr) of the wastewater was around 250 mg L -1 . The microbial 

fuel cells produced a current about 9 mA stably when 249.2±4.6 mg/L 

artificial wastewater as COD including only acetate without nitrate was 

fed. When nitrate from 31.4±0.4 to 123.3±0.1 mg/L was fed into MFCs 

with sodium acetate, the current generation decreased from 9.97±0.06 to 

0.20±0.01 mA according to increase the nitrate concentration. However 

nitrate concentration was removed up to 96% as added nitrate of 123.3±0.1 

mg/L. These results showed that nitrate could behave the electron acceptor 

in the anode compartment. 

Keywords: Microbial fuel cell, nitrate, electricity generation 


405

Bioelectrochemical Fixation of CO2 

Bo Young JEON 1 , Ji-Won CHOI 1 , Il Lae JUNG 2 and Doo Hyun PARK* 1 

1 

Department of Chemical and Biological Engineering, Seokyeong University, 

Seoul 136-704, Korea. 2 J-40 

Department of Radiation Biology, Environmental 

Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon 

305-353, Korea. 


For the isolation of CO2-fixing bacteria using electrochemical reducing 

power (ERP), a plate-type electrochemical bioreactor (PEB) was employed. 

Soluble neutral red (NR) was used as an electron mediator for 

carbonate-basal agar medium (CBAM) prepared in the PEB. For test of 

bacterial CO2 fixation using the ERP, a single-compartmented 

electrochemical bioreactor (SCEB) was employed. NR immobilized in the 

graphite felt cathode (NR-cathode) was used as an electron mediator for 

carbonate-basal broth medium (CBBM) prepared in the SCEB. Two 

bacterial genera capable of electrochemically fixing CO2 were isolated 

using the PEB and cultivated using the SCEB. The isolated bacterial species 

were 99% identified with Achromobacter sp. and Alcaligenes sp. 

Approximate 150 ml of CO2 per day was consumed by 300 ml of the 

mixed culture of Achromobacter sp. and Alcaligenes sp. grown 

electrochemically in the SCEB. Acknowledgement This work was 

supported by the New &Renewable Energy of the Korea Institute of Energy 

Technology Evaluation and Planning (KETEP) grant funded by the Korea 

governmental Ministry of Knowledge Economy (2010T1001100334) 

Keywords: CO2-fixing bacteria, neutral red, electrochemical reducing 

power, Alcaligenes sp., Achromobacter sp. 

Inactivation of Chloramphenicol and Florfenicol by a Novel 

Chloramphenicol Hydrolase from Alluvial Soil Metagenome 

Weixin TAO 1 , Myung Hwan LEE 2 , Jing WU 2 , Nam Hee KIM 2 , 

Jin-Cheol KIM 3 , Eul Chul HWANG 2 and Seon-Woo LEE* 1,2 

1 

Department of Medical Bioscience, Dong-A University, Busan 604-714, South 

Korea. 2 Department of Applied Biology, Dong-A University, Busan 604-714, 

South Korea. 3 J-41 

Chemical Biotechnology Research Center, KRICT, Daejeon 

305-343, South Korea. 

*Corresponding author: Seonlee@dau.ac.kr 

In a previous study, EstDL136 isolated from alluvial soil metagenome 

was redefined as chloramphenicol (Cml) acetate esterase, on account of 

its capability of Cml reactivation that against Cml acetyltransferase (CAT). 

Further investigation of Cml catalytic activity in the absence of CAT 

revealed that Cml was metabolized by EstDL136. The metabolite was 

identified as p-nitrophenylserinol by LC-MS and 1 H-NMR, suggesting a 

promiscuous amidase activity of the enzyme. Purified EstDL136 catalyzed 

Cml hydrolysis when they were co-incubated. Residues were detected by 

HPLC, which showed that Cml was gradually hydrolyzed and the 

hydrolysate, p-nitrophenylserinol was concurrently generated. EstDL136 

also hydrolyzed florfenicol (Ffl), the synthetic florinated analog of Cml 

that was resistant to CAT. Therefore, Escherichia coli was tolerant to 

Cml or Ffl when estDL136 was expressed, and duration of the lag phase 

was highly correlated with the concentrations of supplemented antibiotics 

(Cml, Ffl), indicating that estDL136 mediated the amphenicols resistance. 

This study primarily reported a novel bacterial Cml hydrolase that also 

hydrolyzes Ffl, conferring Cml and Ffl resistance to the E. coli. 

Keywords: Chloramphenicol, Florfenicol, Hydrolase 


J-42 

Efficacy of Combined Application of Entomopathogenic Fungal 

Conidia and Botanical Insecticides for Aphid Control 

Tuan anh PHAM and Keun KIM* 

Dept. of Bioscience and Biotechnology, Univ. of Suwon, Gyeonggi-do 445-743, 

Korea. 

*Corresponding author: ptanh140380@yahoo.com 

Some botanical insecticides prepared from neem, derris, garlic and 

chinaberry were evaluated for the influence of the botanical insecticides 

on susceptibility of conidia of Beauveria bassiana by the germination 

ratio of conidia. The results demonstrated that botanical insecticides 

significantly reduced the germination ratio of conidia and it could not 

be mixed with the conidia in oil-based formulation for long-term storage. 

After that, the virulence of oil-based formulation of conidia in combination 

with botanical insecticides against aphid Myzus persicae was examined 

with detached leaf test. The botanical insecticide was mixed with the 

oil-formulated conidia just before spraying and the result showed that 

the combined application of two insecticides significantly increased the 

mortality of the aphid. 

Keywords: Beauveria bassiana conidia, botanical insecticides, aphid control 

Seasonal Escherichia coli Genotypes Determined by Horizontal 

Fluorophore-Enhanced rep-PCR (HFERP) in the Yeongsan River 

Basin of South Korea 

Jeonghwan JANG 1 , Tatsuya UNNO 1 , Seung Won LEE 1 , Kyung Hwa CHO 1 , 

Michael J. SADOWSKY 3,4 , GwangPyo KO 5 , Joon Ha KIM 1 and 

Hor-Gil HUR* 1,2 

1 


and Technology, Gwangju, Republic of Korea. 2 International Environmental 

Research Center, Gwangju Institute of Science and Technology, Gwangju, Republic 

of Korea. 3 Department of Soil, Water and Climate, University of Minnesota, St. 

Paul, Minnesota, USA. 4 BioTechnology Institute, University of Minnesota, St. Paul, 

Minnesota, USA. 5 J-43 

Department of Environmental Public Health, Seoul National 

University, Republic of Korea. 

*Corresponding author: hghur@gist.ac.kr 

Seasonal and spatial variation in the genotypic diversity of 3,480 Escherichia 

coli isolates obtained from the Yeongsan River basin in South Korea was investigated 

by using the horizontal fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting 

technique. The MDS analysis, done based on HFERP DNA fingerprints, showed 

that E. coli isolates obtained in October through December clustered tightly, while 

those obtained in other sampling periods were more genetically diverse. SOMs 

analysis, done using the 10 most frequently-isolated E. coli genotypes, showed the 

occurrence of season-specific E. coli genotypes and the main SOMs clusters were 

most influenced by temperature, strain diversity, and biochemical oxygen demand. 

Diversity among E. coli genotypes tended to decrease as water temperature decreased, 

and the numbers of E. coli genotypes observed in urban area were greater, more 

diverse, and less dependent on water temperature than those obtained from agricultural 

areas. This work was supported by the National Research Foundation of Korea Grant 

funded by the Korean Government (MEST) (NRF-R1A5A004-2010-0029224), and 

also by the Korea Environmental Technology &Industry Institute (KEITI) as the 

Next Generation Eco Innovation Technology Development Project. 

Keywords: Escherichia coli, Yeongsan River, Horizontal 

Fluorophore-Enhanced rep-PCR

Antifungal Property of Microorganisms against Korea Oak Wilt 

Pathogen, Raffaelea quercus-mongolicae 

Yongsub YI* 1,2 and Jaejoung KIM 2 

J-44 

1 

Department of Herbal Cosmetics Science, Hoseo University, Asan, Korea. 

2 

Department of Biochemistry, Hoseo University, Asan, Korea. 

*Corresponding author: yongsub@hoseo.edu 

This study was performed to investigate the antifungal materials to 

inhibit growth of Raffaelea quercus-mongolicae. Microbes were tested to 

screen antifungal activity. We found microbes showing antifungal activity. 

Two hundreds strains were isolated and incubated on the same fresh media. 

Five strains, SG 1-9, 1-12, SG 2-8, 2-10, and 2-17, were collected by 

determination of antifungal activity. Out of the five strains, organic 

compounds and proteins of SG2-17 strain broth was extracted for 

determination of antifungal activity. The organic compound showed 

antifungal activity, and the proteins extract didn’t show it. The 16S rDNA 

sequences of the above five strains were determined by the sequencing 

analysis. The 16S rDNA sequences were analyzed by the homology of 

BLAST program at NCBI. The homology search showed that SG 1-9 

and 1-12 had 98% homology with Streptomyces cinnamoneus and 98% 

homology with Burkholderia cepacia, SG 2-8, 2-10, and 2-17 had 99% 

homology with Streptomyces fradiae, 97% homology with Staphylococcus 

epidermidis, and 99% homology with Staphylococcus epidermidis. 

Keywords: antifungal activity, Raffaelea quercus-mongolicae 

J-45 

Optimization of the Conditions to make Solid Medium for 

Cyanobacteria 

Seung-Hee SHIN, Mi-Na YU, Eun-Hye PARK, Kyoung-Hee OH and 



361-763, Korea. 


The conditions to make solid medium for cyanobacteria, especially 

focused on the kind and concentration of the solidifying agents, was 

optimized to cultivate these organisms on solid medium, which is 

accounted as the easiest way to establish the axenic culture. Two different 

culture media, modified CB and BG-11, supplemented with bactoagar, 

edible-agar, or agarose, were used. The bactoagar and edible-agar were 

washed with water to remove the potential inhibitors. The concentrations 

of the agents were 0.4% or 1.0%. In order to check the efficiency of 

the culture conditions, the number and diversity of cyanobacteria on the 

solid media were examined. The cyanobacterial numbers on the media 

with rinsed agars were not significantly different from those on the 

unprocessed ones, indicating the inhibitors in these solidifying agents 

cannot be removed with the simple washing. Although the numbers of 

colonies on the BG-11 media were generally higher than those on the 

modified CB media, lots of heterotrophic bacterial colonies were found. 

Based on the number and diversity of cyanobacteria on the media, we 

concluded that the modified CB medium with 0.4% of agarose is the 

adequate condition for solid culture of these organisms. [Supported by 

grants from Eco-Technopia 21 Project] 

Keywords: Cyanobacteria, Solid culture, Agarose 


J-46 

Qualitative Detection of Model Bacteria Expressing Iturin and 

Mop Cyclase and Microbial Community Analyses in Soils During 

the Cultivation of Plants 

Sung Eun KIM, Won-Sik CHOI, Jae Sun MOON and Sung Uk KIM* 

Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, 

Korea. 

*Corresponding author: kimsu@kribb.re.kr 

To detect the microorganisms released into the environment at the 

molecular level, a recombinant strain P. fluorescens MX1 and B. subtilis, 

which have mop cyclase and iturin biosynthetic genes in the chromosome, 

respectively, were employed as model bacteria. Using specific fusion 

primers and the TaqMan probes, quantitative detections of model bacteria 

by real-time PCR were performed using genomic DNAs extracted from 

the soils of cucumber or tomato plants cultivated in pots and the 

greenhouse at intervals of three weeks for a six month period. The genomic 

DNAs from P. fluorescens MX1 and B. subtilis, except for probe DNAs, 

did not show any signals on the Southern blotting, indicating that the 

horizontal gene transfer from model bacteria to soil bacteria did not occur 

during the period monitored in pots and the greenhouse. In addition, the 

T-RFLP patterns of genomic DNAs obtained from the soils of both pots 

and the greenhouse suggest that the changes of microbial communities 

seem to be associated with the seasonal change in the soils. Thus, our 

data suggest that the bacteria used as model microorganisms do not give 

significant impacts on the other bacteria in pots and the greenhouse, 

although a long-term observation may be needed 

Keywords: Real-time PCR, , Horizontal gene transfer, Microbial community 

Effect of Fermentation Conditions on Biohydrogen Production 

from Lipid-rich Food Waste 

Soo Young MOON 1 , Hyo Sun KIM 1 , Can ZHANG 1 , Alma ABUG 1 , 

Sung Chan CHOI 2 and Young Sook OH* 1 

1 

Dept. of Environmental Engineering and Biotechnology, Myongji University, 

Yongin 449-728, Republic of Korea. 2 J-47 

Dept. of Environmental Sciences and 

Biotechnology, Hallym University, Chunchon 200-702, Republic of Korea. 

*Corresponding author: ysoh@mju.ac.kr 

Although carbohydrate, protein, and lipid are basic components of 

organic material, hydrogen yield of carbohydrate fermentation has been 

reported to be significantly higher than that of protein and lipid. This 

study used lard as a model organic matter for lipid, and investigated its 

hydrogen production potential in batch anaerobic fermentation experiments 

under various combinations of stirring and CO2 scavenging condition to 

improve hydrogen production. To optimize hydrogen production, CO2 

scavenging was found to be more effective in lard fermentation compared 

to stirring. In the presence of both CO2 scavenging and stirring, the 

hydrogen yield of lard increased from 4.5 to 185.8 ml-H2/g-VS, and the 

hydrogen content of the total biogas was in the range of 87 - 93% (V/V). 

Acetic, propionic, butyric, and valeric acids were produced during the 

fermentation and valeric acid was found to be the major fatty acid (33.1 

- 67.1%) under all conditions. These results show that hydrogen production 

from lipid materials can be increased by minimizing the partial pressure 

of CO2 and hydrogen and activity of hydrogen-consuming bacteria. 

Keywords: hydrogen production, bioenergy, anaerobic fermentation 


407

Inhibition of Quorum Sensing-Controlled Virulence Factors and 

Biofilm Formation in Pseudomonas aeruginosa by 

2,5-Dimethyl-4-Hydroxy-3(2H)-Furanone (DMHF) 

Can ZHANG 1 , Yu-Hong JIA 1 , Soo-Young MOON 1 , Sung-Chan CHOI 2 and 

Young-Sook OH* 1 

1 

Department of Environmental engineering and Biotechnology, Myongji 

University, Yongin 449-728, Korea. 2 J-48 

Department of Environmental Sciences and 

Biotechnology, hallym University, Chunchon 200-702, Repubic of Korea. 

*Corresponding author: ysoh@mju.ac.kr 

2,5-Dimethyl-4-hydroxy-3(2H)-furanone (DMHF), an innoxious food 

additive, has been proved to have antimicrobial activity recently. However, 

few studies focus on its inhibitory effect on quorum sensing (QS) system 

in pathogenic bacteria. In this study, the production of N-(3-oxododecanoyl)-L-homoserine 

lactone (OdDHL) was completely inhibited by 

1.0 μM DMHF, while N-butyryl-L-homoserine lactone (BHL) production 

decreased on a dose dependent manner by DMHF. 1.0 μM DMHF also 

inhibited QS-controlled virulence factors such as rhamnolipid (55% 

decrease), lasA protease (40% decrease) and pyocyanin (52% decrease), 

and biofilm formation (83% decrease) of Pseudomonas aeruginosa PAO1 

without growth inhibition. However, the production of vi rulence factors 

was restored by the addition of exogenous OdDHL as well as BHL, which 

suggests the inhibition of DMHF is due to the interruption of QS with 

las system as the main target. But the specific inhibition mechanism of 

DMHF on QS system in Pseudomonas aeruginosa remains unknown. This 

study also gives evidence that OdDHL plays more significant role in the 

production of QS-controlled virulence factors than BHL. 

Keywords: Quorum sensing (QS), Biofilm formation, 2, 

5-Dimethyl-4-Hydroxy-3(2H)-Furanone (DMHF) 

The Effect of Treating Rice Straw with Lactobacillus plantarum 

BT-77 and Bacillus subtilis IN-55 on Silage Fermentation 

Hee Jong YANG 2,1 , Jae-Hwa HONG 1,3 , Seong-Yeop JEONG 1 , 

Chan-sun PARK 1 , Hong-gi KIM 3 , Byung-Dae YOON 1 , Yeon-Woo RYU 2 

and Min-Soo KIM* 1 

1 

Bioindustrial Process Center, Korea Research Institute of Bioscience and 

Biotechnology (KRIBB), Jeungeup 580-185, Korea. 2 Department of Molecular 

Science and Technology, Ajou University, san5, Wonchen-dong, Yeongtong-gu, 

Suwon, 443-749, Korea. 3 J-49 

Department of Agricultural Biology, Chungnam 

University, Daejeon 305-764, Korea. 

*Corresponding author: godfilts@kribb.re.kr 

This study aimed to the effects of Lactobacillus plantarum BT-77 and 

Bacillus subtilis IN-55 on manufacturing of rice straw silage. We screened 

the strains having antibacterial and cellulase activity from traditional food 

and soil. Rice straw was treated with control (untreated; TGY broth media), 

BT-77 (4 × 105cfu/g; TGY broth media), IN-55 (4 × 105cfu/g; TGY 

broth media), BT-77 and IN-55 (4 × 105cfu/g; TGY broth media). Under 

optimal conditions (pH 5.5 and 30℃ for 7 days), treatments with BT-77 

increased the concentrations of lactic acid and improved the aerobic 

stability of rice straw. Moreover, BT-77 showed inhibitory activity against 

most of the indicator strains. Additionally, we confirmed not only high 

levels of a cellulase enzyme activity from BT-77 and IN-55, but also 

cell-wall degradation through an electron microscope. In this study, other 

treatments could improve silage quality, but mixed treatment was more 

effective as an additive for rice straw silage. 

Keywords: Rice straw silige, Inoculant, cellulase 


J-50 

Characterization of Manganese Peroxidase Isozyme H4 of 

Phanerochaete chrysosporium Expressed from the yeast Pichia 

pastoris 

Yu-Lim SON, Saravana Kumar KUMAR, Ae-Young MO, Hyun-Young KIM 

and Seung-Moon PARK* 

Division of Biotechnology, College of Environmental and Bioresource Sciences, 

Chonbuk National University, Jeonju, 561-756. 

*Corresponding author: smpark@chonbuk.ac.kr 

Lignin biodegradation is a key step for carbon recycling in terrestrial 

ecosystems, where white-rot basidiomycetes fungi degrade this recalcitrant 

wood polymer enabling cellulose utilization by microbial populations. 

Three peroxidases involved in lignin degradation are known as lignin 

peroxidase (LiP), manganese peroxidase (MnP), versatile peroxidase (VP). 

The cDNA encoding manganese peroxydase isozyme H4 (MnPH4), 

isolated from Phanerochaete chrysosporium, was cloned into the pPICZα 

vectors and expressed in Pichia pastoris under the control of the methanol 

inducible alcohol oxidase I (AOXI) promoter. Enzyme assay indicated that 

the recombinant MnPH4 is efficiently secreted into the medium upon 

hemoglobin supplementation, at a maximum concentration of 500 U l -1 . 

The purified recombinant manganese peroxidase (rMnP) exhibited a 60 

kDa molecular mass considerably larger, because of their glycosylation, 

and showed optimum pH of 4.5 and temperature at 35 o C. 

Keywords: Manganese Peroxidase, Phanerochaete chrysosporium, lignin 

peroxidase 

J-51 

Optimization of the Method to Nucleic Acids from Cyanobacteria 

in the Environmental Samples 

Hye-Ryoung KIM, Eun-Hye PARK, Kyoung-Hee OH and 



361-763, Korea. 


In order to develop the efficient method to extract nucleic acids from 

cyanobacteria in the environmental samples, the concentration and 

disruption methods for cyanobacteria were optimized. Three different 

kinds of filters, polycarbonate, cellulose nitrate, and GF/C, were used to 

concentrate cyanobacteria in the environmental samples. Cyanobacterial 

cells were treated with lysis buffer only or buffer combined with either 

of bead-beating or freezing-thawing. The extraction efficiency was 

examined through quantification of nucleic acids, and the purity was 

checked through PCR or RT-PCR. In case of DNA, the concentration 

with cellulose nitrate membrane and the disruption through bead-beating 

showed the highest extraction efficiency. On the other hand, the use of 

polycarbonate filter and freeze-thawing was adequate methods to extract 

RNA from cyanobacteria. It is needed to figure out why the extraction 

conditions for DNA and RNA were different. The methods developed 

in this study can be helpful to extract nucleic acids efficiently from the 

cyanobacteria in the natural aquatic environments. [Supported by grants 

from National Science Foundation of Korea] 

Keywords: Extraction, Cyanobacteria, Nucleic acid

Characterization of Four Soil Metagenome-Derived Lipolytic 

Enzymes Responsible for Hydrolysis of Autoinducer 

3-Hydroxypalmitic Acid Methyl Ester of Ralstonia solanacearum. 

Myung Hwan LEE 1 , Weixin TAO 2 , Nam Hee KIM 1 , Eul Chul HWANG 1 , 

Jae Wook LEE 3 and Seon-Woo LEE* 1,2 

1 

Department of Applied Biology, Dong-A University, Busan 604-714, Republic 

of Korea. 2 Department of Medical Bioscience, Dong-A University, Busan 604-714, 

Republic of Korea. 3 J-52 

Department of Chemistry, Dong-A University, Busan 

604-714, Republic of Korea. 

*Corresponding author: seonlee@dau.ac.kr 

An autoinducer 3-hydroxypalmitic acid methyl ester (3-OH PAME) an 

unique quorum sensing signal molecule that regulates expression of 

virulence factors such as in phytopathogen Ralstonia solanacearum. 

Previously, we selected 4 metagenomic clones hydrolyzing 3-OH PAME 

among 142 142 lipolytic clones by using gas chromatography. All four 

clones were obtained from industrial waste contaminated soil metagenome 

and designated LP96, ULP104, ULP86 and UDL33. Lipolytic gene from 

LP96B, ULP104 and UDL33BB was over-expressed after cloning in 

pET-30b(+) in Escherichia coli BL21(DE3). The over-expressed, purified 

LP96, LP104 and Est33 exhibited hydrolysis activity toward 3-OH PAME. 

For determining of role of 3-OH PAME hydrolase in quorum sensing 

system of R. solanacearum GMI1000, pRKelp104 as pRK415 carrying 

lp104 gene was transformed into R. solanacearum GMI1000. Growth rate 

and exopolysaccharides (EPS) production of R. solanacearum GMI1000 

carrying pRKelp104 was slower and reducer than R. solanacearum 

GMI1000. These results showed that novel beta-hydroxypalmitic acid 

methyl ester hydrolases were discovered from industrial waste contaminated 

soil metagenome. The enzymes will interfere virulence factors 

expression in R. solanacearum by hydrolysis of autoinducer 3-OH PAME. 

Keywords: Metagenome, 3-OH PAME, beta-hydroxypalmitic acid methyl 

ester hydrolase 

Acute and Chronic Toxicity Assessment for Hazardous Effect 

of Caffeine to Daphnia magna 

Nam Hui HONG 2 , Thai-Hoang LE 2 , Sung-Kyu LEE 3 and Jiho MIN* 1 

1 

Graduate School of Semiconductor and Chemical Engineering, Chonbuk 

National University, 567, Baekje-Daero, Duckjin-gu, Jeonju, Jeonbuk, 561-756, 

KOREA. 2 Department of Bioprocess Engineering, Chonbuk National University, 

567, Baekje-Daero, Duckjin-gu, Jeonju, Jeonbuk, 561-756, KOREA. 3 J-53 

Toxicology 

Research Center, Korea Research Institute of Chemical Technology, Daejon, 

305-600, KOREA. 

*Corresponding author: hongnh@jbnu.co.kr 

Caffeine has been detected in water resulting in the chronic exposure 

of aquatic organisms. Because of organism exposed to hazardousness, 

aquatic ecosystems confusion is coming, and even it affect human. For 

this study, the impact was examined through the acute(48h) and chronic 

toxicity(21d) assay using Daphnia magna as animal model. In the acute 

toxicity test, 10 Daphnia neonates were exposed to several concentration 

of caffeine from 125㎍/l to 4000㎍/l. The lethal concentration with 50% 

mortability (LC50) was determined as about 2225㎍/l. In the chronic 

toxicity test, several small concentrations of caffeine such as 1%, 2%, 

10%, 20% LC50 were used to exposed organism for 21days. We found 

hazardous effect of caffeine on aquatic ecosystem. Although toxicity of 

caffeine was known to be week, we found that survival of Daphnia 

neonates was significantly decreased caused by toxicity of cafferine. Based 

on this result, it was found that aquatic ecosystems exposed by 

pharmaceutics can not be ignored. 

Keywords: Caffeine, Daphnia magna, Acute toxicity, Chronic toxicity. 


Natural Adaptation of Eubacterium limosum KIST612 for Biofuel 

Production 

Shinyoung PARK 1 , Daehee KIM 1 , Sunju CHOI 1 , In Geol CHOI 2 and 

In Seop CHANG* 1 

1 


and Technology (GIST), 261 Cheomdan-gwagiro, Buk-gu, Gwangju 500-712, 

Korea. 2 J-54 

College of Life Sciences and Biotechnology, Korea University, 

Anam-Dong, Seongbuk-Gu, Seoul,136-713, Korea. 


The production of chemicals by microbial reaction has become an 

increasingly alternative to current petroleum-based chemical synthesis. 

Eubacterium limosum KIST612 has been the organism of choice for the 

biochemicals production form hydrogen or carbon monoxide. E. limosum 

KIST612 produces (1) organic acids - acetic and butyric acids, (2) solventethanol 

and (3) gas- carbon dioxide (CO2). Among them, acetate and 

butyrate are major products. We hypothesized that constant transfer in 

certain medium condition this microbe may try to adapt to the given 

environmental condition. We hypothesized that if an excess of acetate 

or butyrate exist with CO as a main electron source, surplus electron 

may be utilized for production of more reduced products.Therefore, the 

hypothesis of this experiment is that excessive acetate and butyrate 

addition into the medium could change metabolic flux. We have tested 

the effect of acetate or butyrate addition for longer than 1 year with CO 

as a main energy source. We found significant increase of solvents, ethanol 

and butanol production after 1 year adaptation. It was also clearly observed 

that acetate could be converted to more reduced products in the presence 

of excess CO. 

Keywords: Acetogen, Eubacterium, Adaptation 

J-55 

Mortality of Moth Larvae by Various Entomopathogenic Fungi 

Young-Hoon LIM, Eun-Soo GO and Keun KIM* 

Dept. of Bioscience and Biotechnology, The University of Suwon, Gyeonggi-do 

445-743, Korea. 

*Corresponding author: limyounghoon@korea.com 

To determine the virulence of the various fungi, 10 8 conidia/㎖ 

suspension of each fungal strain was sprayed onto 2rd instar of larvae 

on a dampened filter paper in a petri-dish and the petri-dish was incubated 

at 25℃ for 15 days. The mortality by the different fungal strains varied 

from 20% to 100%. Among various fungi tested, 4 strains showed relativity 

high mortality against larvae. The mortality of the larvae increased and 

the 50% lethal time(LT50) decreased significantly as the applied conidial 

concentration was raised. LT50 values of strain No. 198, 216, 218, and 

500 against larvae were 12.7, 9.5, 8.4, and 9.3 days, respectively. Mean 

lethal concentration(LC50) values of strain No. 198, 216, 218 and 500 

against the larvae were 3.62 × 10 7 , 9.94 × 10 6 , 6.97 × 10 4 , and 3.01 

× 10 5 conidia/㎖, respectively. 

Keywords: Entomopathogenic Fungi, Mortality, Moth Larvae 


409

J-56 

Development of Baeyer-Villiger Monooxygenases Screening 

Protocols Using High-throughput screening (HTS) system 

Gowoun LEE, Yunjon HAN, Jae Jun SONG and Jong Hyun CHOI* 

Enzyme-based Fusion Technology Research Team, Microbe-based Fusion 

Technology Research Center, Korea Research Institute of Bioscience and 

Biotechnology (KRIBB), 1404 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea. 

*Corresponding author: jhchoi@kribb.re.kr 

Baeyer-Villiger Monooxygenases (BVMOs) are flavoenzymes that 

catalyze the insertion of an oxygen atom in a C-C bond using NADPH 

and allow the conversion of (cyclic)-ketones into lactones or esters. For 

the development of BVMO screening protocol, we used fluorescence based 

substrate and robot based HTS system. Optimized BVMO screening 

protocol for high throughput screening system led to 6,000 clone active 

verification per a day was available. As result, we screened the new type 

of BVMO positive clone which is named as “GomsoMF” from "Gomso" 

metagenomic library. This “GomsoMF” was novel BVMO like enzyme 

can convert cyclohexanone to ε-caprolactone. Also this enzyme was 

convert cyclopentanone and cycloheptanone into δ-valerolactone and 

7-heptanolactone respectively. This new BVMO will be used for the 

synthesis of useful biochemicals or the biotechnological application. (This 

work was supported by a high throughput screening (HTS)-based 

Integrated Technology Development grant from the MEST, and a basic 

research grant from KRIBB.) 

Keywords: Baeyer-Villiger Monooxygenases (BVMOs), high throughput 

screening 

J-57 

The Effect of Gamma Irradiation on Poliovirus, a Model for 

Norovirus inoculated in Oyster (Crassostrea Gigas) and Effect 

of Culture Condition 

Jong-Il CHOI, Eu-Ri JO, Pil-Mun JUNG, Jae-Hun KIM, Beom-Suk SONG, 

Jae-Kyung KIM, Jong-Heum PARK and Ju-Woon LEE* 

Team for Radiation Food Science and Biotechnology,Advanced Radiation 

Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185. 

*Corresponding author: sjwlee@kaeri.re.kr 

Poliovirus is a recognized surrogate for norovirus, pathogen in water 

and food, due to the structural and genetic similarity. Although radiation 

sensitivity of poliovirus in water or media had been reported, there has 

been no research in food model such as shellfish. In this study, oyster 

(Crassostrea gigas) was incubated in artificial seawater contaminated with 

poliovirus, and thus radiation sensitivity of poliovirus was determined in 

inoculated oyster. The effects of ionizing radiation on the sensitivity of 

poliovirus were also evaluated under different conditions such as pH (4-7) 

and salt concentration (1-15%) in culture broth, and temperature during 

irradiation. The D10 value of poliovirus in PBS buffer, virus culture broth 

and oyster was determined to 0.46, 2.84 and 2.94 kGy, respectively. The 

initial plaque forming unit (PFU) of poliovirus in culture broth was slightly 

decreased as the decrease of pH and the increase of salt concentration, 

but radiation sensitivity was not affected by pH and salt contents. However, 

radiation resistance of poliovirus was increased at frozen state. These 

results provide the basic information for the inactivation of pathogenic 

virus in foods by using irradiation. 

Keywords: Radiation sensitivity, Poliovirus, Oyster 


Application of Bacillus subtilis 168 to Improve Durability of 

Cement 

Sung-jin PARK 1 , Jong-Myong PARK 1 , Wha-Jung KIM 2 and 

Sa-Youl GHIM* 1 

J-58 

1 

Dep. of Microbiology, Kyungpook National University, Daegu 702-701, Korea. 

2 

Dept. of Architecture Engineering, Kyungpook National University, Daegu 

702-701, Korea. 

*Corresponding author: ghimsa@knu.ac.kr 

Calcium carbonate precipitation, a widespread phenomenon among 

bacteria, has been investigated due to its wide range of scientific and 

technological application in concrete remediation. In this study, Bacillus 

subtilis 168 which could induce calcium carbonate precipitation, was used 

to coat on the cement surface. CaCO3 crystal aggregates and organic matrix 

molecules of these strains were deposited on the surface of the cement 

paste. To verify the effect of coating on the surface of the cement paste, 

water permeability of the paste was measured by water immersion test. 

In the results, B. subtilis 168 addition showed lower water permeability 

than those treated with B4 medium only, suggesting that the precipitated 

calcium induced by B. subtilis 168 could be used as a sealing agent to 

block water penetration. Also, the calcium precipitation on the surface 

of cement mortar could increase the compressive strength of the mortar. 

The compressive strength test showed that the cement mortar treated with 

B. subtilis 168 showed an improvement of compressive strength. 

Keywords: Calcium carbonate precipitation, Bacillus subtilis 168, cement 

J-59 

Screening Enzyme from Single cell Based Polymerase Fosmid 

Cloning 

Yunjon HAN, Kyong-Cheol KO, Jong Hyun CHOI and Jae Jun SONG* 

Enzyme-based Fusion Technology Research Team, Microbe-based Fusion 

Technology Research Center, Korea Research Institute of Bioscience and 

Biotechnology (KRIBB), 1404 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea. 

*Corresponding author: jjsong@kribb.re.kr 

"Metagenomics," which is the genomic analysis of the microbial 

community attempts to overcome culturability of various microbes. In 

general, the diversity of metagenomic libraries was affected by abundance 

of bacteria in environmental sample. Therefore the research of minor 

microbial texa in environmental samples remains the biggest challenge. 

A new method was developed for enrichment minor bacteria from 

environmental samples. This method is based on the Fluorescence in situ 

hybridization(FISH), Fluorescence associated cell sorter(FACS) and 

Multiple displacement amplification(MDA). We demonstrated enrichment 

minor bacteria from artificial microbial community and single cell based 

MDA followed by fosmid library construction for enzyme screening. The 

Jeongupia naejangsanesis in which the cellulolytic activity is known was 

used in the minor microorganism of the artificial microbial community. 

As a result, we obtained MDA products from single bacterial cell. And 

we could generate fosmid library having an average insert size of 32 

kb. Among this fosmid library we screened two new cellulose genes. (This 

work was supported by a high throughput screening (HTS)-based 

Integrated Technology Development grant from the MEST, and a basic 

research grant from KRIBB.) 

Keywords: Single cell, Multiple displacement amplification, Screening 

enzyme

Virulence of Entomopathogenic Fungus Lecanicilium sp. Le85 

towards Cotton Aphid 

Van Hanh VU* 1 , Dinh Thi QUYEN 2 , Xuan Dat VU 2 , Huu Quan NGUYEN 2 

and Keun KIM 3 

1 

Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi 

10600, Vietnam/ Department of Bioscience and Biotechnology, The University of 

Suwon, Hwaseong-si 445-743, Korea. 2 Institute of Biotechnology, Vietnam 

Academy of Science and Technology, Hanoi 10600, Vietnam. 3 J-60 

Department of 

Bioscience and Biotechnology, The University of Suwon, Hwaseong-si 445-743, 

Korea. 

*Corresponding author: vvhanh@ibt.ac.vn 

Twenty-two entomopathogenic fungal strains were used for laboratory 

bioassay against aphid Aphis gossypii living on cucumber leaves. Among 

them, four strains of Lecanicillium spp. isolated from infected aphid’s 

cadavers had high virulence towards A. gossypii causing 84 to 100% 

mortality after 5 to 7 days of conidial treatment. Especially, one fungus 

Lecanicillium sp. Le85 showed the highest virulence towards development 

and reproduction of A. gossypii. At 20~24 o C and relative humidity of 

75~85%, after 5 days of application with 1x10 8 conidia of Le85 /ml, A. 

gossypii did not develop and exhibited a mortality of 100% with LT50 

of 2.6 days. Moreover, at 25~31 o C and relative humidity of 60~70%, 

the mortality of A. gossypii by Le85 was about 95 ~100% after 5 days 

with LT50 of 2.8 days. These results suggested the potential role of Le85 

for use as microbial control agent against A. gossypii in greenhouses. 

Keywords: Cotton aphids Aphis gossypii, entomopathogenic fungi, 

Lecanicillium sp. 

J-61 

Partial Identification of Microorganisms from Artificially 

Constructed Tunnel and Dokdo Soil to Develop Antifungal 

Construction Material 

Jong-Myong PARK, Sung-Jin PARK and Sa-Youl GHIM* 

School of Life Sciences, Kyungpook National University, Daegu 702-701, Korea. 

*Corresponding author: eveningwater@hanmail.net 

To develop antifungal concrete, we sampled microorganisms from the 

tunnel to determine the most widely distributed fungal strain. In addition, 

we are finding the multi-functional (antifungal and calcite forming) 

bacterium from the Dokdo soil. Isolation and pure culture of bacterium 

and fungi were done using potato dextrose agar and several bacteriological 

media.As the result of partial identification, fungal strain showed the 

closest homologies with Penicillium sp., Tritirachium sp., Elsinoaceae sp., 

Cladosporium spp., Alternaria spp., Pestalotiopsis cocculi, Pseudozyme 

prolifica or Aureobasidium pullulans, respectively. Each bacterial strains 

showed the closest similarity with Rhodococcus erythropolis (99.794%), 

Bacillus aryabhattii (99.791%) or Bulkholderia glathei (96.369%), 

respectively. The most widely distributed fungal strain was Cladosporium 

spp. As the result of the calcite forming assay, 52 Dokdo derived bacterial 

strains and five tunnel derived strains were identified as calcite forming 

bacteria (CFB). Qualitative antimycotic assay showed that 23 Dokdo and 

two tunnel derived CFB strains have antimycotic activity against 

Aspergillus niger KCTC6906 and Cladosporium spp. using the strain's 

crude liquid culture. 

Keywords: Antifungal, Construction, Material. 


Novel Esterases Isolated from Marine Sediment Metagenome 

So-Hyeon SEO 1 , Sang-Hong YOON 1 , Young-Seok LEE 1 , 

Sung-Gyun KANG 2 , Jung-Hyun LEE 2 and Chang-Muk LEE* 1 

1 

Functional Biomaterial Division, National Academy of Agricultural Science, 

Rural Development Administration, Suwon 441-707, Korea. 2 J-62 

Korea Ocean 

Research and Development Institute, Ansan 426-744, Korea. 

*Corresponding author: changmuk@rda.go.kr 

Metagenomes from uncultured microbial communities are rich sources 

for identifying novel biocatalysts. With deep sea sediments at an island, 

Dokdo, we have constructed the metagenomic libraries comprising total 

60,672 fosmid clones. A fosmid clone with lipolytic activity was selected 

by functional screening of the metagenomic libraries using tributyrin agar 

plates. To identify genes, a shot-gun library was constructed from the 

selected clone by random shearing. Further functional screening of the 

shot-gun library revealed one novel clones. The sequence analysis of the 

clone revealed the presence of an open reading frame encoding acidic 

polypeptide of 59.4kDa (553 amino acids, pI of 4.45) with the highest 

homology to a carboxyl esterase from δ-proteobacterium Haliagium 

ochaceum DSM 14365(44% identity, 61% similarity). The sequence 

analysis showed that the protein harbors the typical catalytic triad formed 

by Ser229, Asp347 and His461. Multiple sequence alignments with known 

carboxyl esterases showed the clone belongs to the carboxylesterase BioH 

member, which is involved in biotin biosynthesis in Aromatoleum 

aromaticum [This experiment is supported by grants from the Rural 

Development Administration project number PJ0067032011 and IPET 

project number 110037-03-1-HD110]. 

Keywords: carboxylesterase, metagenome, dokdo 

K_ Host-Microbe Interactions 

K-1 

Aphids Feeding Manipulates Host Plant Immunity against Plant 

Pathogens 

Choong-Min RYU* and Soohyun LEE 

Laboraytory of Bio-industry and Biochemistry Research Center, KRIBB 111 

Gwahangno, Yuseong-gu, Daejeon 305-806, S. Korea. 

*Corresponding author: cmryu@kribb.re.kr 

Plants modulate defense signaling networks to response against different 

biotic stresses. In this work, we evaluated whether insect infestation can 

change of plant defense mechanism charging to subsequent pathogen 

attack. Aphids and/or a chemical trigger benzothiadiazol (BTH) was 

pretreated seven days before challenging two pathogenic bacteria, 

Xanthomonas axonopodis pv. vesicatoria (Xav) as a compatible pathogen 

and X. axonopodis pv. glycines 8ra (Xag 8ra) as an incompatible pathogen. 

Disease severity was noticeably lower in aphid and BTH + aphid 

treatments than control. BTH or aphids alone treatment did not differ 

induction of hypersensitive response (HR) against Xag 8ra. However, the 

combination treatment showed synergistically to reduced HR appearance. 

Pretreatment of BTH resulted in significant reduction of aphids population. 

Expression of the two defense-related genes, Capsicum annum 

Pathogenesis-related genes 1 (CaPR1) and CaPR4 revealed that aphid 

infestation primes resistance genes resulting in induction of systemic 

defense response against compatible and incompatible pathogens. Taken 

together, our results suggest that insect feeding itself elicits propound plant 

resistance and help plant coping subsequent attack of pathogens. 

Keywords: Aphid, immunity, Xanthomonas 


411

J_Environmetal Microbiology and Engineering

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