I_Food Microbiology and Biotechnology

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12 Φεβ 2013 (πριν από 4 χρόνια και 4 μήνες)

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I
_ Food Microbiology and Biotechnology
Antioxidant Action of Reaction Mixtures of
Mono-O-α -D-Galactopyranosyl-4-O-β -D-Mannopentaose
and Amino Acids
Gwi Gun PARK*
2
and Chang Yeop LEE
2
1
Dept. of Food &Bioengineering, Kyungwon University.
2
Dept. of Food
&Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
Mono-O-α -D-galactopyranosyl-4-O-β -D-mannopentaose separated
and identified from enzymatic hydrolysate to gums by the purified β
-mannanase from Xylogone sphaerospora. Although coloration was low
at reaction mixture of mono-O-α -D-galactopyranosyl-4-O-β
-D-mannopentaose from Xylogone sphaerospora and tyrosine, methionine,
and cysteine, but TLC pattern of reaction mixture of mono-O-α
-D-galactopyranosyl-4-O-β -D-mannopentaose and tyrosine appeared new
reaction mixture spot having the different Rf value. The browning reaction
solution, obtained by heating an aqueous solution (0.1M HCl-99.8%
ethanol, 1:1) of amino acids (tyrosine, methionine, and cysteine) at 100

for 8hrs, were tested with coloration, TLC, reducing power, antioxidant
activity and DPPH(2, 2-Diphenyl-1-picrylhydrazyl) test.
Keywords: galactosyl mannooligosaccharide, browning reaction
I-1
Quality and Microbiological Characteristics of Commercial
Long-Term Fermented Baechu Kimchis
Jhon Deok Young JHON
1
, Yun LEE
2
and Deok Young JHON*
1
1
Dept of Food and Nutrition, Chonnam National University, Gwangju, 500-757,
Republic of Korea.
2
Dept of Food and Nutrition, Graduate School, Chonnam
National University, Gwangju, 500-757, Republic of Korea.
*Corresponding author: dyjhon@gmail.com
Quality and microbiological characteristics of five commercial
long-term fermented Baechu Kimchis were investigated. Levels of
moisture, ash, crude protein and crude fat were identified. Contents of
reducing sugar, total vitamin C, and organic acids were also determined.
In color measurement, L, a and b values of the Kimchis were 51.1

65.3,
13.4

28.3 and 26.9

30.6, respectively. Total viable bacteria, lactic acid
bacteria and yeasts in the Kimchis were 1.0×10
5

5.0×10
6
CFU/g, 3.5×10
4

1.2×10
6
CFU/g and 9.0×10
4

1.7×10
6
CFU/g, respectively. Hardness
values (18,048

23,253 g/cm
2
) were not significantly different among the
commercial products (p<0.05). Based on overall acceptability scores of
the Kimchis, predominant lactobacilli and yeasts were isolated and
identified from two commercial products, 1K and 2S, using an API system.
L. brevis (4.6×10
5
CFU/g) and C. maltaromaticum (1.4×10
5
CFU/g) were
major lactobacilli in 1K and L. brevis (2.4×10
5
CFU/g) and
L
.
mesenteroides ssp. mesenteroides/dextranicum (1.1×10
5
CFU/g) were
predominant microorganisms in the Kimchi, 2S. Major yeast strains were
identified as C. guilliermondii (5.5×10
3
CFU/g) and K. lodderi (1.6×10
3
CFU/g) in 1K. In 2S, all the identified strains were S. unisporus (5.8×10
5
CFU/g).
Keywords: long-term fermented baechu kimchis, commercial kimchi,
identification of lactic acid bacteria and yeasts
I-2
Construction and Expression of E. coli-Lactic Acid Bacteria
Shuttle Vectors, pGYC3α and pGYC4α
Hae Choon CHANG* and Eun Ju YANG
Dept. of Food and Nutrition, Chosun University, Gwangju, Korea.
*Corresponding author: hcchang@chosun.ac.kr
The plasmid pYC2 in Lactobacillus sakei isolated from kimchi, was
used to construct E. coli-LAB shuttle vectors. The shuttle vector, pGYC3
α, was constructed using pYC2 replicon, E. coli ColE1 replication origin,
erythromycin resistance gene, and α -amylase gene of
B
acillus
licheniformis as a food-grade selection marker. In addition, pGYC4α and
pGYC5α were constructed containing P32 promoter, or P32 and usp45
signal peptide of Lactococcus lactis for expression of a foreign protein,
α -amylase. The three shuttle vectors were transformed successfully into
E. coli and several lactic acid bacteria, which were isolated from kimchi.
α -Amylase activities were detected in E. coli, Lb. sakei,
L
euconostoc
mesenteroides harboring pGYC3α, and in E. coli, L. lactis,
L
euconostoc
citreum, Leu. mesenteroides harboring pGYC4α. But the transformants
harboring pGYC5α vector did not show any α -amylase activities. The
segregational stability result indicated that the constructed shuttle vectors
were significantly stable, although they have a rolling-circle replicon from
pYC2 plasmid.
Keywords: Lactobacillus sakei, shuttle vector, α -amylase
I-3
Fermentation of Banana Peel (Musa acuminata, AA Group
cv. Lacatan)
Noime JACINTO, Jae-Ho CHO, Min-Jeong PARK, Dae-Il HWANG,
Sung-Il CHO, Seung-Yong SHIN and Tae-Wan KIM*
Department of Food Science and Biotechnology, Andong National University,
Andong 760-749, Korea.
*Corresponding author: jacinto_noime@yahoo.com
Banana is one of the staple foods in the Philippines. It is also used
as an ingredient in most of the food industry thereby creating large amount
of industrial waste in the form of a peel. The goal of the present work
was to evaluate the properties of banana peel from Lacatan (
M
usa
acuminata, AA Group) using various fermentation microorganisms such
as Bacillus strains (B1, B2, BL1, BL2, BL3, B. licheniformis) and LAB
strains (A2, A3, A4, A5, L. acidophilus, Coopers, Wills) for 0, 24 and
48 hour basis. Extracts were evaluated for its protein, total sugar, reducing
sugar, total Phenolic compounds. Result showed 755, 802, 758, 790, 757,
804 mg/g protein content on the 3
rd
day of fermentation using Bacillus
strains which is almost three times higher compared with the protein
content using LAB strains However, total Phenolic compounds of peel
fermented with LAB strains showed a significantly higher amount
compared with Bacillus strains, the highest being L. acidophilus (144 mg/g,
3
rd
day of fermentation) which is almost four times higher compared with
the highest content in Bacillus strains (BL1, 41 mg/g). Likewise, reducing
and total sugar were higher in LAB strains fermentation. A4 showed the
highest value on the 3
rd
day of fermentation, 203 mg/g and 116 mg/g
(A5) respectively.
Keywords: lacatan, Bacillus, lactic acid bacteria
I-4
345
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Evaluation of Proantocyanidin Contents and Antioxidative
Activities of Various Pine (Pinus densiflora) Needle Extracts
Sang-Hyeon LEE*
1
, Nam-Young KIM
1
, Ki Hwan YU
1
, Min-Kyung JANG
1
,
Hye Ji JANG
1
, Yong Soo PARK
2
, Min Hee JEON
2
, Young Kyoung KIM
2
,
Mihyang KIM
2
, Sung Gu KIM
3
and Byung Hong YOO
3
1
Dept. of Pharmaceutical Engineering, Silla University, Busan 617-736,
K
orea.
2
Dept. of Food and Nutrition, Silla University, Busan 617-736, Korea.
3
Bioport
Korea Co., Busan, 619-912 Korea.
*Corresponding author: slee@silla.ac.kr
We evaluated proanthocyanidin contents in total polyphenolic compounds
of pine needle extracts prepared by ethanol, hexane, ethanol-sub-supercritical
(ESS) or finely ground powder-sub-supercritical (FGPSS). Analyses of total
polyphenolic compounds and proanthocyanidin in each extracts indicated that
ethanol extract contained the highest concentrations, but FGPSS extract
contained the lowest concentrations. On the other hand, evaluation of
proanthocyanidin contents in total polyphenolic compounds in each extracts
showed that hexane extract possessed the highest content, but FGPSS extract
possessed the lowest content. These results indicate that extracts containing
high concentrations of both total polyphenolic compounds and
proanthocyanidin could be obtained by using ethanol or hexane extraction.
Analyses of DPPH and FIC of extracts showed that ethanol and hexane extracts
possessed the highest activities. FGPSS extract showed the best FRAP where
as ESS extract showed the lowest DPPH and FRAP activities. In this study,
we prepared extracts from pine needles by four different methods and evaluated
the antioxidative compounds in extracts. Acknowledgement: This work was
support by Human Resource Training Project for Regional Innovation, Ministry
of Knowledge Economy, Korea.
Keywords: pine needle, antioxidative activity, proanthocyanidin
I-5
Antimicrobial Activity of Sargassum fulvellum Ethanol Extract
So Young YOON
1
, So Young LEE
1
, Koth Bong Woo Ri KIM
1
,
Eu Jin SONG
1
, Seo Jin KIM
1
, So Jeong LEE
1
, Chung Jo LEE
1
,
Na Bi PARK
1
, Ji Yeon JUNG
1
, Ki Wan NAM
2
and Dong Hyun AHN*
1
1
Dept. of Food Science &Technology, Pukyong National University, Busan
608-737, Korea.
2
Dept. of Marine Biology, Pukyong national university, Busan
608-737, Korea.
*Corresponding author: dhahn@pknu.ac.kr
Antimicrobial activity of Sargassum fulvellum(SF) was investigated
b
y
paper disc assay and MIC test, and inspected stability of processing
characteristics through extreme heat and pHs. Additionally silica gel
column chromatography and HPLC were performed in order to identify
major antimicrobial component of SF. The result of measured
antimicrobial effect, SF ethanol extract showed great antimicrobial activity
agaisnt gram positive bacteria by paper disc assay at level of 4 mg/ml.
The MIC value of SF ethanol extract range from 0.05 to 0.0063% against
test microbes. At the heat and pH stability test, the result suggest that
SF ethanol extract is greatly considered the suitable substance in change
of heat and pH. SF ethanol extract was fractionated to hexane, chloroform,
ethylacetate, butanol and water. As a result of MIC test, the chloroform
fraction showed the strongest level of antimicrobial activity among the
different five fractions. The chloroform fraction was separated using silica
gel column chromatography, and 16 of sub-fractions were obtained.
Among them, CH4 which showed the highest activity purified by HPLC
and we obtained three main peak. In oder to describe the mechanism
of SF, we experiment on time kill curve, loss of 260nm absorbing material
and microscopy.
Keywords: antimicrobial activity, identify, mechanism
I-6
Anti-inflammatory Effects of Fermented Beans Through
NF-κ B Activation
Hea Hyeon LEE
1
, Cheol PARK
2
, Min Jeong KIM
1
, Min Jeong SEO
1
,
Yung Hyun CHOI
3,4
, Cheol PARK
2,4
and Yong Kee JEONG*
1
1
Department of Medical Bioscience, Dong-A University, Busan 604-714, South
Korea.
2
Department of Biochemistry, Dongeui University College of Oriental
Medicine.
3
Department of Biochemistry, Dongeui University College of Oriental
Medicine, Busan 614-714, South Korea.
4
Department of Biomaterial Control
(BK21 Program), Dongeui University Graduate School, Busan 614-714, South
Korea.
*Corresponding author: rmflsql1@nate, com
Cyclooxygenase (COX)-2 is generally known as an inducible enzyme, and
it produces arachidonic acid to prostaglandin E2 (PGE2), which have
b
een
demonstrated to play critical roles in inflammation. In present study, we
investigated the effects of the extracts of fermented beans including soybean
(FS), black agabean (FBA) and yellow agabean (FYA), on the expression o
f

COXs and production of PGE2 in human leukemic U937 cell model. Treatment
of phorbol 12-myristate 13-acetate (PMA) significantly induced
p
ro-
inflammatory mediators such as COX-2 and PGE2 production, whereas the
levels of COX-1 remained unchanged. However, pre-treatment with FS, FBA
and FYA significantly decreased the PMA-induced COX-2 protein as well
as mRNA, which was associated with inhibition of PGE2 production. Moreover,
FS, FBA and FYA markedly prevented the increase of nuclear translocation
of nuclear factor kappa B (NF-kB) p65 by PMA. Our data indicate that the
extracts of fermented beans exhibits anti-inflammatory properties
b
y
suppressing the transcription of pro-inflammatory cytokine genes through the
NF-kB and EGR-1 signaling pathway.
Keywords: FS, FBA, FYA
I-7
Isolation and Characterization of Promoters from Leuconostoc
mesenteroides SY2
Ji Yeong PARK
2
, Ae Ran LEE
1
, Kang Wook LEE
2
, Ji Yeon LEE
2
,
Hwang A LEE
2
, Chang Un BAEK
2
, Hyeon Deok JO
2
, Joo Yeon KIM
2
and
Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
*Corresponding author: jeonghkm@gsnu.ac.kr
Leuconostoc mesenteroides has been regarded as the most dominant and
important organism during the early and middle stages of kimchi fermentation.
Development of expression vectors for Leuconostoc species is important for
the basic and applied researches on kimchi LAB. The chromosomal DNA o
f

Leuconostoc mesenteroides SY2 was digested with Sau3AI and then cloned
into the unique BamHI site of pBV5030, which contains a promoterless cat-86
encoding a chloramphenicol(Cm) resistance gene. Sixteen fragments were
identified based on their ability to confer Cm resistance in E. coli. Plasmids
contaning the clones were designated as pJY series plasmid. The length of
each fragment ranged from 0.3 to 1.3 kb. The exact sizes of the fragment
exhibiting promoter activity were 499, 644, 756, 979 and 1330 bp for 10C,
18C, 22C, 25C and 34C, respectively. The maximum Cm resistance observed
for most of these clones reached up to 1500 μ g/ml. Expression vectors with
powerful promoters could be useful for the industrial production of valuable
proteins in LAB such as lactobacilli and leuconostoc species. J. Y. Park, K.
W. LeeJ. Y. Lee, H. A. Lee, and C. U. Baek were supported by the 2nd
stage Brain Korea 21 program from the Ministry of Education and Human
Resources Development, Republic of Korea
Keywords: Leuconostoc mesenteroides, kimchi LAB, promoter
I-8
346
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Microbiological Analysis of Commercial Sprouts
Sunggi HEU*, Jin-hyuk SON, So Yeon PARK, Eun Kwang LIM,
Eunjung ROH, Sam Nyu JEE, Kyu Suk JUNG and Doo-Ho KIM
Microbial Safety Division, National Academy of Agricultural Science, RDA,
Suwon 441-707.
*Corresponding author: heu@rda.go.kr
Recently sprouts became very popular for “Bibimbab” and salad in
Korea. Sprouts grown for food are baby plants that are harvested just
after germination. A large number of microorganisms are able to
proliferate on sprouts. Five the most common crop seed sprouts, cabbage,
radish, alfalfa, broccoli, and rape, were analyzed for
b
acterial
contaminants. The total aerobic bacteria count was approximately 6~9
log CFU/g. Total aerobic bacteria count was relatively higher than that
of leafy greens. Since most commercial sprouts packages contained leafy
greens for decoration, the bacterial contaminants were analyzed for leafy
greens with sprouts. The presence of seven different pathogenic
b
acteria
was analyzed. Results using selective media showed the possible presence
of most pathogenic bacteria. However, most pathogenic bacteria were not
detected by PCR and Biochemical test except S. aureus from decorative
leafy greens in sprouts packages. S. aureus contamination was found in
more than 16% of leafy green samples and showed contaminations of
more than 3 Log CFU/g. Since these results showed that the sprouts
production processing is relatively safe but the some decorative leafy
greens were contaminated with S. aureus, it may be better to pack sprouts
without any decoration in it.
Keywords: Staphylococcus aureus, sprout
I-9
Characterization of Alcohol Fermentation Saccharomyces sp.
Isolated from Wild Grape (Vitis coignetiea)
Sung Yeon CHOI
1
, Jung Bok LEE
2
, Chun Pyo JEON
1
, Chung Sig CHOI
3
,
Oh Seuk LEE
4
, Kee Sun SHIN
5
and Gi Seok KWON*
1
1
School of Bioresource Sciences, Andong National University, Andong 760-749,
Korea.
2
Dept. Food &Nutrition Science, Kundong University, Andong 760-833,
Korea.
3
Bio Industry Institute, HansBio Co., Ltd. Andong 760-380, Korea.
4
D
ept.
of Food Fermentation Technology, Youngdong University, Youngdong 370-701,
Korea.
5
Biological Resource Center, Korea Research Institute of Bioscience and
Biotechnology (KRIBB), Daejeon 305-600, Korea.
*Corresponding author: gskwon@andong.ac.kr
In this study, For the development of wild grape wine, we investigated
its optimum fermentation conditions as well as quality changes during
fermentation. The isolated strain HSC 10 was identified as Saccharomyces
cerevisiae. This strains showing a high tolerance against osmotic pressure
in the presence of 50% glucose, high tolerance to alcohol(10%) and
potassium metabisulfite tolerance(500

/

). The wild grape wine
fermented with S. cerevisiae HSC 10 at 30 °Brix juice and 20

showed
characteristics in terms of alcohol production and titratable acidity. The
glucose contents of the wild grape wine drastically decreased and with
fermentation time. Alcohol contents of S. cerevisiae HSC 10 was 13.2%
after 9 days fermentation. Antioxidant activity and SOD-likely activity
of the wild grape wine maintained 85

86% and 29

30% during
fermentation for 15days, respectively. But, Total polyphenol contents
increased during fermentation for 15days. The results suggested that wild
grape wine was brewed by fermentation of 15days at 20

by S. cerevisiae
HSC 10 has the potential to become a novel functional wild grape wine
with good acceptability.
Keywords: alcohol fermentation, wild grape, Saccharomyces cerevisiae
I-10
Enhanced Production of Ultra High Molecular Weight
Poly-γ -Glutamic Acid (UHMWPGA) from Bacillus subtilis
Chungkookjang
Yoon-Ho CHOI
1
, Byoung-Chan LEE
1
, Hyoung-Bum KIM
1
, Sung-Jin LEE
1
and Moon-Hee SUNG*
1,2
1
BioLeaders Corporation, Daejeon 305-500, Korea.
2
Dept. Bio &Nanochemistry,
Kookmin University, Seoul 136-702, Korea.
*Corresponding author: smoonhee@bioleaders.co.kr
Poly-γ -glutamic acid (γ -PGA) is an extracellular amino acid polymer
as a major adhesive component of fermented soybeans. The molecular
weight of γ -PGA range from 10 kDa to 10,000 kDa, dependent on γ
-PGA producer. Primarily, we established that the Bacillus subtilis
chungkookjang BLS isolated from Chungkookjang, a traditional fermented
soybean food, produces ultra high molecular weight γ -PGA
(UHMWPGA) in the industrial scale. Then we established the enhanced
production of UHMWPGA (5,000kDa above) using the fermentation
medium and tricarboxylic acid cycle intermediates on the production of
g-PGA(13~18g/L, high molecular weight analysis basis). The maximum
recovery yield of 10~12g/L was obtained with high molecular weight,
e.g. the 5,000kDa polymer using the 5KL jar fermentor system. We
developed the industrial production process of UHMWPGA. [This work
was supported by the 'Seoul R&BD Program(10580)’]
Keywords: ultra high molecular weight (UHMW), poly-γ -glutamic acid,
industrial production
I-11
Microbial Spoilage of Minimally Processed Vegetables
Sunggi HEU*, So Yeon PARK, Jin Hyuk SON, Eunjung ROH,
Sam Nyu JEE, Se-Ri KIM and Doo-Ho KIM
Microbial Safety Division, National Academy of Agricultural Science, RDA,
Suwon 441-707.
*Corresponding author: heu@rda.go.kr
Almost 30% of vegetables were discarded by the spoilage from farms
to tables. After harvest, vegetables are often spoiled by a wide variety
of microorganisms including many bacterial and fungal species. The
diversity of microbes related to the spoilage of vegetables had
b
een
investigated. The total aerobic bacterial numbers in lettuce, perilla, and
cichori were 2.7

10
6,
4.6

10
5,
1.2

10
6,
respectively. The most
b
acterial
species were Pseudomonas SP., Pectobacterium SP., and
B
urkholderia
SP. and other 17 more species were involved in. After 1 week of incubation
of those vegetables at 25

, the microbial diversity had been changed.
The total aerobic bacterial numbers increased to 4.6

10
8,
4.9

10
7,
and
7.6

10
7
for greens that were about 10
2
times more bacterial numbers
than that before spoilage. However, the diversity of microbes isolated
had been simplified. Fewer bacterial species had been isolated and
characterized. The most bacterial population (~60%) was taken up
b
y
Pseudomonas sp. including P. fluorescence, P. viridiflava, and P.
marginalis and followed by Arthrobacter sp. and Bacillus sp. The spoilage
activity of bacterial isolates had been tested using axenic lettuce plants.
Among tested isolates, P. fluorescens and Pantoea agglomerans cased
severe spoilage on lettuce.
Keywords: spoilage of vegetables, Pseudomonas sp.
I-12
347
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Effects of the Addition of Carbohydrates on the Isoflavone
and Oligosaccharide Contents in Soymilk During
Fermentation with Weisella sp. 4.
Ji-Yeon CHUN
2
, Farhana SHARMIN
2
, Woo Ju JEONG
1
, Kang Wook LEE
1
and Jeong Hwan KIM*
1
1
Gyeongsang National University.
2
Sunchon National University.
*Corresponding author: cjyfall@sunchon.ac.kr
Soymilk was fermented with Weissella sp.4 producing β -glucosidase
in order to prepare functional soy-yoghurt containing high level of
isoflavone aglycones. Three carbohydrates including glucose, sucrose and
starch (2%, w/v) were added to soymilk and changes in viable cell number,
acid production, the content of isoflavones and oligosaccharides (raffinose,
stachyose) were investigated during fermentation at 37

for 12 h. Viable
cell numbers of Weissella sp.4 in soymilk rapidly increased from 10
-6
CFU/g to 10
-9
CFU/g. During fermentation, pH of 6.3 rapidly dropped
into the range of 4.3~4.5 and titratable acidity increased from 0.15% upto
0.55%. There was no significant difference in those levels among three
fermented soymilks containing glucose, sucrose and starch. About 98%
of isoflavone glucosides were hydrolyzed in all soymilks during
fermentation. Hydrolysis rates of isoflavone glucosides were slightly
improved by the addition of glucose and sucrose. The levels of raffinose
and stachyose slowly decreased in all soymilks during fermentation.
Soymilk with glucose showed the lowest raffinose content at the end of
fermentation. WJ Jeong were supported by the BK 21 program from the
Ministry of Education &Human Resources Development, Republic of
Korea.
Keywords: isoflavone, fermentaion, soymilk
I-13
Antibacterial test of Bacillus polyfermenticus KJS-2 against
Some Bacteria Isolated from Rotifer and Artemia Culture
Aquarium
Kang Min KIM
2
, Min Hee PARK
2
and Jae Seon KANG*
1
1
Department of Pharmacy, Kyungsung University, Busan, 608-736,
K
orea.
2
Department of Smart Foods and Drugs, Inje University, Gimhae, Gyeongnam,
621-749, Korea.
*Corresponding author: kkij79@hanmail.net
The Bacillus polyfermenticus KJS-2, an endospore-forming rod, was
first isolated by Dr. Jae Seon Kang in 2006. Shewanella abalonesis,
Arthrobacter nicotianae, Vibrio parahaemolyticus and Psychrobacter
nivimaris were isolated by rotifer and artemia culture aquarium. The
identification of isolated bacteria was performed with 16S rDNA
sequencing from Korean Culture Center of Microorganisms. Antibacterial
activities of two isolated pathogenic bacteria were tested by top-agar
method using Bacillus polyfermenticus KJS-2. Bacillus polyfermenticus
KJS-2 showed a highly antimicrobial activity against two pathogenic
bacteria. The strong inhibition activities of Bacillus polyfermenticus KJS-2
were observed for two pathogenic bacteria.
Keywords: Bacillus polyfermenticus KJS-2, rotifer, artemia
I-14
Development of Non-added Sugar Wine with Domestic
Campbell's Early
Oh Seuk LEE*, Wongook JUNG and Chul YOOK
Dept. of Wine &Fermented Food, Youngdong University, Chungbuk 370-701,
Korea.
*Corresponding author: los25@daum.net
Domestic Campbell's Early contains insufficient sugar content to
b
rew
wine. In generally we are brew wine with a method called "Chaptalization"
that adding sugar at early stage of fermentation. The wine(made
b
y
"Chaptalization") is insufficiently international competitive because strong
sour taste and weak flavor. This investigation tried to solve the problem
with new method that dried grapes by a warm current air(40

),
investigated changes of physicochemical properties during fermentation.
As a result, the wine A(produced by a new method) contains 16% alcohol,
the wine B(produced by “Chaptalization”) contains 12% alcohol. The wine
A contains 1.12% total acidity, the wine B contains 0.78% total acidity
as tartarate. The wine A showed 13.8 whiteness, the wine B showed 56.9
whiteness, the wine A showed 38.9 redness, the wine B showed 55.8
redness, the wine A showed 23.7 yellowness, the wine B showed 11.9
yellowness respectively. So we thought that the wine A is closer deep
red color. The wine A is much more contains polyphenolic pompound
with 1.30 mg/mL than the wine B contains 0.88 mg/mL polyphenolic
compound. We considered as manufacturing a new characteristic
Non-added wine that it contains higher polyphenol content and alcohol
content and sugar content in Korea.
Keywords: wine, none-added sugars, campbell's Early
I-15
Comparison of Inhibitory Effect of Three Different Classes
of Bacteriocins on Lactic Acid Bacteria from Kimchi
Min Ho CHOI*
1
, Chul Ho KIM
1
and Yun Hee PARK
2
1
Molecular Bioprocess Research Center, Jeonbuk Branch Institute, KRIBB, 1404
Sinjeong-dong, Jeongeup 580-185, Korea.
2
Department of Molecular Science and
Technology, Ajou University, San 5 Woncheon-dong, Suwon.
*Corresponding author: choimh@kribb.re.kr
Kimchi is a traditional Korean food, consisting of vegetables fermented
by lactic acid bacteria. It is difficult to control the fermentation of kimchi,
because it is a spontaneous and incomplete process involving several
species of lactic acid bacteria. Therefore, the most important problem
facing the kimchi industry is to develop an effective method of
fermentation control thereby facilitating preservation. In this study, we
investigated the inhibitory effect of three different types of
b
acteriocins,
nisin, pediocin PA-1, and enterococcin EFS2, against lactic acid
b
acteria
from kimchi. Three bacteriocins showed different inhibitory patterns with
respect to antimicrobial spectrum and effect of environmental conditions;
pH, NaCl, and temperature. Especially, nisin showed the highest activities
and widest inhibitory spectrum against Lactobacillus spp., responsible for
over-acidification of kimchi. Moreover, the inhibitory effect of nisin was
enhanced by NaCl and at low pH while it decreased in pediocin PA-1
and enterococcin EFS2. Therefore, nisin could be effectively used to
control lactic fermentation of kimchi usually containing 3% NaCl and
having high acidity.
Keywords: bacteriocin, kimchi, inhibitory effect
I-16
348
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r

Characterization of Bacteriocin Produced by Lactococcus
lactis ET-45 Isolated from Kimchi
Seong Yoep JEONG, Nack-Shick CHOI, Chan-Sun PARK,
Keug-Hyun AHN, Hee-Jong YANG, Byung-Dae YOON and Min-Soo KIM*
Jeonbuk Branch Institute Bioindustry Research Center, KRIBB, Jeon buk 580-185,
Korea.
*Corresponding author: khs8706@paran.com
Bacteriocin producing lactic acid bacteria were isolated from kimchi
using a MRS selective plate with Bacillus cereus as an indicator strain.
16S rDNA sequence and sugar utilization test identified that ET-45 was
a Lactococcus lactis strain. The bacteriocin exhibited inhibitory activity
against Bacillus cereus, Leuconostoc mesenterides,
L
euconostoc
carnosum, Leuconostoc lactis, Leuconostoc mesenteroides
s
ubsp.
cremoris, Lactobacillus plantarum, Lactobacillus acidophilus,
Lactobacillus viridesceus, Pediococcus dextrinicus, and Enterococcus
f
acecium. Optimal production of the bacteriocin was obtained by growing
the cells on MRS medium at pH 7.5 and 30

for 18~24 hrs. Sucrose
and protease peptone are essential for bacteriocin production as carbon
source and nitrogen source, respectively. Antibacterial activity of the
bacteriocin was completely disappeared by proteinase K, which indicates
it’s proteinous nature. The bacteriocin was inactivated by protease such
as trypsin, α -chymotrysin, Subtilisin A. The bacteriocin was fully stable
at 121

for 60 min. Solvents such as chloroform, ethanol, acetone,
acetonitrile, hexane, isopropanal did not effect on the activity.
Keywords: bacteriocin, Lactococcus lactis, kimchi
I-17
Physicochemical Properties of Wine Produced by
Thermovinification with Domestic Campbell's Early
Oh Seuk LEE*, Kangsu JEON, Wongook JUNG and Chul YOOK
Dept. of Wine &Fermented Food, Youngdong University, Chungbuk 370-701,
Korea.
*Corresponding author: los25@daum.net
Domestic wine(produced by "chaptalization" that adding sugar at early
stage of fermentation) had a weak point that insufficient color especially
deep red color with the consequence to be insufficiently international
competitive. This investigation tried to solve the problem with
"thermovinification" that first extraction heat treatment in a double
boiler(80

) for half hour, investigated changes of physicochemical
properties during fermentation. As a result, the wine A(produced
b
y
"thermovinification") contains 8.4% alcohol, the wine B(produced
b
y
"chaptalization") contains 6.5% alcohol. The wine A contains 0.90% total
acidity(as tartarate), the wine B contains 0.73% total acidity. The wine
A is more contains polyphenolic compound with 1.64mg/mL than the
wine B contains polyphenolic compound with 0.18mg/mL. The wine A
is more closer deep red color as a chromaticity investigation that the wine
A showed 9.52 whiteness, the wine B showed 23.66, the wine A showed
43.08 redness, the wine B showed 54.31 redness, the wine A showed
1.75 yellowness, the wine B showed 22.87 yellowness respectively. This
investigation thought that we can brew a characteristic wine that enriched
color produced by "thermovinification".
Keywords: thermovinification, domestic grape, polyphenolic compound
I-18
Physicochemical Characteristics during Fermentation of
Kochujang Prepared with Wheat Flour
Song Yi HAN, Min Hwa KIM, Jin Bo JEONG and Yong Suk KIM*
Department of Food Science &Technology, Chonbuk National University,
Jeonbuk 561-756, Korea.
*Corresponding author: kimys08@chonbuk.ac.kr
Changes in physicochemical characteristics of Kochujang prepared with
3 starch source, glutinous rice (GK), domestic wheat flour (DWK), and
imported wheat flour (IWK), were investigated during fermentation of
10 weeks at 25

. Moisture contents of all treatments were continually
increased during fermentation. pH of GK, DWK, and IWK were gradually
deceased from 4.88, 4.86 and 4.97 at the initial stage to 4.78, 4.75, and
4.83 after 10 weeks, respectively. In contrast, total titratable acidities were
increased to 0.86, 0.46 and 0.64% at the initial stage to 1.15, 0.77, and
0.90% after 10 weeks. Salt contents of GK, DWK, and IWK were
maintained at level of 10, 7, and 7%, respectively, during fermentation.
After 10 weeks. Amino-type nitrogen contents of IWK showed the higher
123.91 mg% than those of GK (114.80 mg%) and DWK(120.87 mg%).
the color of all treatments was not changed during fermentation, but that
of GK was lower than those of DWK and IWK. Major free sugars of
all treatments were glucose, maltose and fructose, and citric, malic,
succinic and acetic acid was detected organic acids.
I-19
Accumulation of Ergothioneine in Gastrodia elata Rhizomes
Eung-Jun PARK*
1
, Wi Young LEE
1
, Jin Kwon AHN
1
, Seung Taek KIM
2
1
Divi. of Forest Biotechnology, Korea Forest Research Institute, Suwon 441-847,
Korea.
2
Dept. of Horticulture, Kyunghee University, Yongin 446-701, Korea.
*Corresponding author: pahkej@forest.go.kr
Gastrodia elata must establish mycorrhizal association with symbiotic
fungi such as Armillari gallica for its vegetative growth. Erogothioneine
(ERG), which is an excellent antioxidant, was accumulated in A. gallica
at the different levels in fruiting bodies (1.95 mg/g DW), rhizomorphs
(0.32 mg/g DW), and mycelia (0.15 mg/g DW), respectively. Rhizomes
(tubers) of G. elata also accumulated ERG of which concentration was
significantly different depending on their size and environmental condition.
The smaller rhizomes generated by in vitro culture accumulated
approximately 10-fold higher amount of ERG (~ 10.17 mg/g DW)
compared to that in the larger rhizomes (1.06 mg/g DW) produced in
cultivation areas. Histochemical analysis revealed that the fungal infection
area was restricted in the cortical cells, indicating that the ERG contents
were correlated with the surface area of G. elata rhizomes. Furthermore
the antioxidant capacity was significantly correlated with the ERG
concentrations in the G. elata rhizomes, as measured by DPPH, ABTS,
and ORAC
hydro
. In the present study, we found that the rhizomes of G.
elata contained ERG from 0.1% to 1% of their dry weight, suggesting
that ERG is another major component with free radical-scavenging
activities in G. elata. .
Keywords: Gastrodia elata, Armillari gallica, erogothioneine
I-20
349
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Quality characteristics of DJI Doenjang made with Bacillus
subtilis Starter and Solar Salt
Hae Choon CHANG* and Mi CHANG
Dept. of Food and Nutrition, Chosun university, Gwangju, 501-759, Korea.
*Corresponding author: hcchang@chosun.ac.kr
Bacterial-koji and Doenjang were prepared using B. subtilis DJI and
refined salt or solar salt with concentration of 12%(w/v). According to
the increase of aging period(up to 2 months), moisture contents(%) of
refined salt-DJI Doenjang and solar salt-DJI Doenjang were decreased
from 51.55, 51.88 to 45.38, 47.90, respectively. Acidity of these DJI
Doenjang was gradually increased. Hunter‘s L(lightness) color values of
the Doenjangs were decreased. Browning reaction rate of refined salt-DJI
Doenjang was faster than solar salt-DJI Doenjang during 2 months aging
period. The compositions of total and free amino acids and their ratios
of the Doenjangs were changed during the aging period. The fibrinolytic
activities of DJI Doenjangs, traditional Doenjangs and commercial
Doenjangs were 827.71~885.14 units/ml, 47.30~479.73 units/ml and
250.00~418.92 units/ml, respectively. Flavor compounds of DJI Doenjang
and traditional Doenjang were examined by SDE and GC/MS. Typically
favorable flavor compounds in traditional korean Doenjang; 2, 3,
5-trimethylpyrazine, benzaldehyde and 2, 5-dimethylpyrazine were
detected from DJI Doenjang with reduced off-flavor compounds. We
believe that this study is the first report about easy and rapid Doenjang
manufacture method by using bacterial-koji system.
Keywords: bacterial-koji, doenjang, Bacillus subtilis DJI
I-21
Isolation of Bacteria from Onion Fermentation for the
Development of Feed Additive
Woo-Kyung CHANG
1
, Xiao-Tian QUAN
1
, Jun-Hyeong LEE
1
,
Sang-Buem CHO
1
, Dong-Woon KIM
2
and Soo-Ki KIM
1
*
1
Department of Animal Science and Technology, Konkuk University, Seoul
143-701, Korea.
2
National institute of Animal Science, RDA
*Corresponding author: sookikim@konkuk.ac.kr
In this study, to develop fermented onion as feed additives, various
strains of bacteria were isolated from fermented onion. LB, MRS, PDA
and YM broth with onion juice were naturally fermented or with the
addition of soil supernatant. Eight colonies were isolated from naturally
fermented onion and thirty two colonies were from soil inoculation. From
natural fermentation, Lactobacillus sp. and Bacillus sp. were identified
by 16S rDNA sequencing. Enterobacter sp., Pseudomonas sp.,
Lactobacillus sp., Burkholderia sp., and Delftia sp. were predominately
identified from 32 colonies that isolated from the fermentation with soil
inoculation. In the analysis of digestive enzyme activity, most high
protease was found in strain SK2086. SK2006(Enterobacter sp.),
SK2009(Burkholderia sp.) and SK2011(Delftia sp.) showed high amylase
activities and SK2003(Weissella sp.) represented high CMCase activity.
Lipase activity was detected in SK1996(Burkhoderia sp.), SK1997,
SK1998, and SK2009(Burkholderia sp.). Antibacterial activities against
animal pathogenic bacteria Burkholderia sp. and Heamophillus sp. were
found in Lactobacillus sp. SK2090 and SK2091. With further
investigations, the isolated strains will be effectively used for onion
fermentation.
Keywords: onion, fermentation, feed additives
I-22
Development of a RAPD-PCR Method for Identification of
Bacillus Species Isolated from Chunggukjang
Ji Yeon LEE
2
, Ae Ran LEE
1
, Kang Wook LEE
2
, Ji Yeong PARK
2
,
Hwang A LEE
2
, Chang Un BAEK
2
, Hyeon Deok JO
2
, Joo Yeon KIM
2
,
Kun Young PARK
3
and Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
3
Department of Food Science
and Nutrition, Pusan National University, Busan 609-735.
*Corresponding author: jeonghkm@gsnu.ac.kr
Bacillus cereus is a food-borne spore-forming pathogen that is ubiquitous
in the natural environments. To establish a rapid and accurate method for
the specific detection of B. cereus from natural environments including
fermented foods, we performed RAPD-PCR for B. cereus standard strains in
30 μ L reaction volume. All B. cereus strains tested produced common
b
ands
of 0.5 and 0.95 kb in size. The 0.5 kb band was gel extracted and sequenced.
It turned out to be a part of a ytcP gene encoding a hypothetical
p
rotein
similar to a ABC-type transporter. We previously showed that the same 0.5
kb fragment was observed in all B. subtilis strains. The sequence of 0.95 kb
band observed in B. cereus strains did not match to any known sequences
in the database. The RAPD-PCR procedure was used to identify B. cereus
isolates from chunggukjang and three isolates showed the unique pattern of
B. cereus. recA gene sequencing confirmed that they were indeed B. cereus.
The results showed the usefulness of the RAPD-PCR method for rapid
identification of B. cereus from fermented foods. JY Lee, HA Lee, JY Park
and CU Baek were supported by the 2nd stage Brain Korea 21 program from
the Ministry of Education and Human Resources Development, Republic of
Korea.
Keywords: RAPD-PCR, Bacillus cereus
I-23
The Biotransformation of the Flavonoids: Curcumin,
Phloretin, Apigenin, Chrysin Using Various Microorganisms
Junehyung KIM*
1
, Jong-Ki LEE
1
, Jiwon ROH
1
, Woo-Il KIM
1
,
Kwonyoung CHOI
2
, TaeJin KIM
2
, Byung-Gee KIM
2
1
Department of Chemical Engineering, Dong-A University, Busan, Korea.
2
School
of Chemical and Biological Engineering, Seoul National University, Seoul,
K
prea.
*Corresponding author: june0302@dau.ac.kr
Flavonoids are polyphenolic compounds that are ubiquitous in nature
and are categorized according to chemical structure, into flavonols,
flavones, flavanones, isoflavones, catechins, anthocyanidins and chalcones.
Over 4,000 flavonoids have been identified, many of which occur in fruits,
vegetables and beverages (tea, coffee, beer, wine and fruit drinks). The
flavonoids have aroused considerable interest recently because of their
potential beneficial effects on human health-they have been reported to
have antiviral, anti-allergic, antiplatelet, anti-inflammatory, antitumor and
antioxidant activities. Microorganisms are well known for their abilities
to catalyze many types of useful biotransformation reactions with
flavonoids including hydroxylation and methylation.
Keywords: flavonoid, biotransformation, curcumin
I-24
350
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r

Antimutagenic Effect of Gentiooligosaccharides from
Aspergillus niger β -Glucanase
Gwi Gun PARK* and Meung Seok LEE
Dept. of Food &Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
This study was performed to elucidate antimutagenic effect using
gentiooligosaccharides from curdlan by Aspergillus niger β -glucanase.
The composition of medium for Aspergillus niger was 2% peptone, 0.5%
yeast extract and incubated for 144hr at 35

and 180rpm.
Gentiooligosaccharides were separated quantitatively by activated carbon
column chromatography. Aspergillus niger β -glucanase was partial
purified by DEAE Sephadex column chromatography. The β -glucanase
exhibited maximum activity at pH 6 and 60

. Among the three kinds
of metals(Cu2+, Mg2+, Fe2+), only Fe ion broke DNA at the concentration
of above 5mM. In the DNA breaking test, the gentiooligosaccharides
recovered broken DNA in the presence of Fe2+.
Keywords: DNA breaking test
I-25
Cloning and Characterization of a Fibrinolytic Enzyme of
Bacillus amyloliquefaciens CH86-1, Isolated from
Chunggukjang
Ae Ran LEE
1
, Kang Wook LEE
2
, Ji Yeon LEE
2
, Ji Yeong PARK
2
,
Hwang A LEE
2
, Chang Un BAEK
2
, Hyeon Deok JO
2
, Joo Yeon KIM
2
and
Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
*Corresponding author: jeonghkm@gsnu.ac.kr
Bacillus amyloliquefaciens CH86-1 with strong fibrinolytic activities was
isolated from Chunggukjang and it produced several proteases. Among four
different growth media, LB broth was the best in terms of cell growth and
fibrinolytic activities. From culture supernatant, a protein with fibrinolytic
activity was partially purified by CM-Sephadex and Phenyl-sephadex column
chromatographies. Tandem mass analysis results showsd that it was a homolog
of AprE from B. subtilis and the protein was accordingly named AprE86-1.
The optimum pH and temperature for partially purified AprE86-1 were 7.0
and 40℃, respectively. A fibrinolytic enzyme gene, aprE86-1, encoding a 27
kDa enzyme was cloned from the genomic DNA of CH86-1, and the DNA
sequence determined. aprE86-1 was overexpressed in heterologous B. subtilis
strains deficient in extracellular proteases using a E. coli-Bacillus shuttle vector,
pHY300PLK. A 27 kDa AprE86-1 band was observed and AprE86-1 seemed
to exhibit higher activities towards fibrin. HA Lee, JY Park, JY Lee, and
CU Baek were supported by the 2nd stage Brain Korea 21 program from
the Ministry of Education and Human Resources Development, Republic of
Korea
Keywords: Bacillus amyloliquefaciens, chunggukjang
I-26
Organophosphorus Hydrolase (opd gene) of Lactobacillus
brevis WCP902 from Kimchi is able to Degrade
Organophosphorus Pesticide
Kye Man CHO
3
, Renukaradhya K MATH
1
, Shah Md. Asraful ISLAM
1
,
Jong Min KIM
1
, Myoung Geun YUN
1
, Ji Joong CHO
1
, Eun Jin KIM
1
,
Weon Taek SEO
3
, Hoon KIM
4
and Han Dae YUN*
1,2
1
Division of Applied Life Science (BK21 Program), Gyeongsang National
University, Chinju 660-701, Korea.
2
Research Institute of Agriculture and Life
Science, Gyeongsang National University, Chinju 660-701, Korea.
3
D
epartment
of Food Science, Jinju National University, Chinju 660-758, Korea.
4
D
epartment
of Agricultural Chemistry, Sunchon National University, Suncheon 540-742,
Korea.
*Corresponding author: hdyun@nongae.gsnu.ac.kr
We isolated Lactobacillus brevis WCP902 that is capable of
biodegrading chlorpyrifos from kimchi. The opdB gene cloned from this
strain revealed 825 bp, encoding 274 aa, and an enzyme molecular weight
of about 27 kDa. OpdB contains the same Gly-X-Ser-X-Gly motif found
in most bacterial and eukaryotic esterase, lipase, and serine hydrolases,
yet it is a novel member of the GDSVG family of esterolytic enzymes.
Optimum organophosphorus hydrolase (OpdB) activity was appeared at
pH 6.0 and 35

and during degradation of CP, CM, DZ, MPT and PT.
Its conserved serine residue, Ser82, is significantly involved with enzyme
activity that is supposed to be used to remove some pesticide contaminants
from raw materials of kimchi [Supported by a grant No.
R01-2008-000-20220-0 from the Basic Research Program of KOSEF].
Keywords: pesticide-degrading bacterium, Lactobacillus brevis, opdB
gene
I-27
Production of Glyceollins in Different Varieties of Soybean
Challenged by Various Fungal Species
Ae Ran LEE
1
, Mee Ryung LEE
1
, Kang Wook LEE
2
, Ji Yeon LEE
2
,
Ji Yeong PARK
2
, Hwang A LEE
2
, Chang Un BAEK
2
, Hyeon Deok JO
2
,
Joo Yeon KIM
2
and Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
*Corresponding author: jeonghkm@gsnu.ac.kr
Glyceollins have anticancer and antimicrobial activities. In this study,
we investigated the glyceollin induction in different variety of soybeans
by 5 fungla species; Aspergillus niger, Aspergillus oryzae,
A
spergillus
sojae, Aspergillus awamori and Rhizopus oligosporus. Differens varieties
of soybeans (Taeg-gwang, A-ga, cheng-ja, Dae-yaung, Nam-pung, and
Sun-chang) were inoculated with each fungal spores after soaked in water
and autoclaving. Glyceollin (I, II. And III) amount was measured
b
y
HPLC. Overall, R. oligosporus infection caused efficient induction of
glyceollins in all the soybean varieties tested. When three different
methods of fungal inoculation into soybean were compred, sample of
half-cut soybean gave much higher glyceollin contents than those of
soybean chopped, and soybean homogenized. The results indicated that
maintaining biosynthetic capability of soybean was important in induction
of glyceollin synthesis. Glyceollin rich soybean might be useful for the
production of various soy foods such as soy milk and soy yogurt. HA
Lee, JY Park, JY Lee, and CU Baek were supported by the 2nd stage
Brain Korea 21 program from the Ministry of Education and Human
Resources Development, Republic of Korea
Keywords: glyceollins, soybean
I-28
351
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Antioxidant Activities of Fermented Aga Beans Ethanolic
Extracts
Min Jeong KIM
1
, Min Jeong SEON
1
, Hye Hyeon LEE
1
, Woo Hong JOO
2
and Yong Kee JEONG*
1
1
Department of Medical Bioscience, Dong-A University, Busan 604-714,
K
orea.
2
Department of Biology, Changwon National University, Changwon, 641-773,
Korea.
*Corresponding author: min-Jung722@hanmail.net
Fermented normal soybean (FS), yellow Aga bean (FYA) and
b
lack
Aga bean (FBA) exhibited good fibrinolytic activities of 0.88, 1.17 and
1.25 unit/ml, respectively. FYA extract showed an highest DPPH radical
scavenging activity at 15~82%, until the concentrations of 0.01~1 mg/ml,
while FS and FBA at 11~76%, respectively. FBA exhibited highest another
antioxidative activities such as SOD-like activity. Both FYA and FBA
were showed significantly higher antioxidative activities than that of FS.
The antioxidative activities of three extracts were increased in a
dose-dependent manner, until the concentrations of 0.01~1 mg/ml with
increasing of concentration.
Keywords: FS, FYA, FBA
I-29
Storage Improvement of Salted Mackerel by Korean Herbal
Extracts Treatment
Min-Jeong PARK, Ju-Young HWANG, Noime JACINTO, Dae-il HWANG,
Se-Hui JO, Hyun-Ae KIM and Tae-Wan KIM*
Department of Food Science and Biotechnology, Andong National University,
Andong 760-749, Korea.
*Corresponding author: booooj@nate.com
This study was carried out to analyze the quality change of salted
mackeral treated with extracts of Sanguisorba officinalis L. (SOE),
Syzgium aromaticum (SAE), Rosmarinus officinalis L. (ROE) and Alpinia
officinarum L. (AOE) during storage for development of high quality
mackerel. Total plate counts of the salted mackerel non-treated and treated
herbal extracts ranges from 5.6 to 9.8×104 CFU/g at 0 day of storage.
Especially, it was lower at 7 days of storage in AOE-treated salted
mackerel among herbal extracts compared with control (salted mackerel
not treated with herbal extracts). The content of trimethylamine (TMA),
the substance mainly responsible for the fish odor, was analyzed by using
HPLC. TMA contents of salted mackerel until 3 days were not detected
with a value of <0.02 but was significantly increased as the term went
on. However, TMA values in all herbal extracts treated salted mackerel
was 1.5 mg/Kg and 10 times lower than that of control at 7 days of
storage. The overall acid values (AV) in all herbal extracts treated salted
mackerel during storage were significantly decreased compared with
control. The AV of AOE-treated salted mackerel was 5.852 mg/g and
it was lower than that of control (10.032 mg/g) at 7 days of storage.
Keywords: salted mackerel, herb, trimethylamine
I-30
Functional Mechanism of α -Galactosidase from Mortierella
vinacea to Galactosyl Mannooligosaccharides Having the
High/Low D.P Value
Gwi Gun PARK*
2
and Eun Jeong SEO
2
1
Dept. of Food &Bioengineering, Kyungwon University.
2
Dept. of Food
&Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
α -Galactosidase from Mortierella vinacea liberated galactose from
melibiose, raffinose, stachyose by TLC. Locust bean gum was hydrolyzed
D.P. 5(Gal3Man4), 7(Gal2, 3Man5) by β -mannnanase from Bacillus sp.,
D.P. 4(Gal2Man3), 7(Gal2Man6) by β -mannnanase from Trichoderma.
harzianum and D.P. 4(Gal2Man3), 6(Gal2Man5) by β -mannnanase from
Xylogone sphaerospora. These hydrolysates were separated by activated
carbon coulmn chromatography. The enzyme acted on D.P. 4, 5 of
galactosyl mannooligosaccharides from Bacillus sp., Trichoderma
harzianum and Xylogone sphaerospora sphaerospora. But the enzyme
didn't on D.P. 6, 7 of galactosyl mannooligosaccharides from Bacillus
sp., .Trichoderma harzianum and Xylogone sphaerospora
Keywords: Mortierella vinacea, α -galactosidase
I-31
Microbial and Componential Characteristics of Korean
Traditional Soybean Fermented Food; Chungkookjang
Ji-Young BYUN
1
, Sung-Bin PARK
2
, Jae-Su MOON
2
, Mi-Sun KWAK
2
and
Moon-Hee SUNG*
1,2
1
BioLeaders Corporation, Daejeon, Korea.
2
Dept. Bio &Nanochemistry,
K
ookmin
University, Seoul, Korea.
*Corresponding author: smoonhee@bioleaders.co.kr
Chungkookjang is a Korean traditional fermentation soybean containing
microorganisms, enzymes, and diverse bioactive compounds, which are
absent in unfermented soybean. Chungkookjang is also called Natto,
Tempeh, and Douchi in other Asian regions and can be prepared in a
short period without using sodium chloride, which is different from the
preparation of other Korean fermented soybean pastes. We have studied
the microbial and componential characteristics of chungkookjang. We
suggest that it is mainly depend on microorganisms, used for fermentation.
Raw chungkookjang has only one strains of microorganism,
B
aciilus
subtilis or Bacillus licheniformis for fermentation. But we confirmed that
common chungkookjang has many strains. Therefore, common
chungkkokjang smells more than raw chungkookjang. And we identified
that chungkookjang was only fermented by Bacillus species, because we
did not find Lactobacillus species in chungkookjang. And we investigated
the content of γ -PGA and the molecular weight of γ -PGA, and so on.
In this studies, we confirmed that the quality of chungkookjang is
depended on microorganisms and the quality of chungkookjang could
increase by using sole microorganism when fermentation. [This work was
supported by the 'Seoul R&BD Program(10580)’]
Keywords: poly-γ -glutamic acid, Korean traditional fermented food,
chungkookjang
I-32
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r

Antioxidative Ability of Lactic Acid Bacteria
Chul-Ho LEE*, Young-Tae AHN, Jung-Hee LEE and Chul-Sung HUH
R &D Center, Korea Yakult Co., Ltd., 418-12, Korea.
*Corresponding author: cenalch@re.yakult.co.kr
Nine strains of lactic acid bacteria were investigated for antioxidative
activity. These included Lactobacillus plantarum HY7711, AK8-3, K8-1a,
and K12-2; Lactobacillus sp. KY2-8, KM6-2, KM7-8 and KM8-5. Intact
cells of all strains demonstrated antioxidative activity, showing anpha,
α -diphenyl-β -picrylhydrazyl (DPPH) free radical scavenging activity
b
y
20-37%. The reducing activity of intact cells and intracellular cell free
extracts of all strains were studied. Most of the strains tested demonstrated
excellent reducing activity. Lactobacillus plantarum HY7711 had the
highest reducing activity. The Oxygen Radical Absorbance Capacity
(ORAC) assay was also performed on all strains. The ORAC value was
expressed as micromole Trolox equivalent per 10
9
CFU.
L
actobacillus
plantarum HY7711 also showed the highest ORAC value. The results
of this study indicate that lactic acid bacteria have antioxidative activity.
Among the 9 strains tested, Lactobacillus plantarum HY7711 is the most
capable of scavenging reactive oxygen species and possessing reducing
activity. In addition, lactic acid bacteria may be potential candidates for
production of natural antioxidant supplements.
Keywords: lactic acid bateria, antioxidative ability, ORAC
I-33
Production of 2, 6-Dimethoxy-p-Benzoquinone from Wheat
Germ Using Saccharomyces cerevisiae Isolated from Wine
Eun-Hee PARK, Mi-Hee JANG, Jong-Gil YOO and Myoung-Dong KIM*
School of Bioscience and Biotechnology, Kangwon National University,
Chuncheon 200-701, Korea.
*Corresponding author: mdkim@kangwon.ac.kr
β -Glucosidase often plays a crucial role for production of
aglycone-type functional material from glycone-type, less-effective
substrate. Saccharomyces cerevisiae strains isolated from wine were
examined for their β -glucosidase activities for production of
aglycone-type of 2, 6-dimethoxy-p-benzoquinone (2, 6-DMBQ), an
anticancer compound, from wheat germ. Among the 44 tested strains,
S. cerevisiae KCTC17592 yielded highest extracellular β -glucosidase
activity of 0.02 U/mg protein to produce 2, 6-DMBQ with a content of
0.02 mg/g substrate, which corresponded to over two-fold increase in 2,
6-DMBQ content compared with that obtained from usual laboratory S.
cerevisiae strain BY4742.
Keywords: aglycone, 2, 6-dimethoxy-p-benzoquinone, β -glucosidase
I-34
Isolation and Characterization of Bacillus am
y
loli
q
ue
f
aciens
with Antifungal and Antibacterial Activities from Meju and
Cheonggukjang
Hwang A LEE
2
, Ji Yeong PARK
2
, Kang Wook LEE
2
, Ji Yeon LEE
2
,
Ae Ran LEE
1
, Chang Un BAEK
2
, Hyeon Deok JO
2
, Joo Yeon KIM
2
and
Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
*Corresponding author: jeonghkm@gsnu.ac.kr
Bacilli with proteolytic activities were screened from Meju, and ganjang,
Korean traditional fermented soyfoods. Total 448 strains with protease activities
were isolated and among them, 4 isolates were selected for antimicrobial
activities.The isolates were named MJ1-4, MJ3-6, MJ5-48, and MJ7-66,
respectively. Previously isolated B. amyloliquefaciens CH86-1 and B.
amyloliquefaciens CH51 strains were also examined for the antimicrobial
activities. Among them MJ1-4 showed the most strong inhibition against a
variety of fungi and CH51 against a variety of bacteria. Supernatant of MJ1-4
and CH51 cultures were concentrated by 80% ammonium sulfate
p
recipitation
and the concentrated supernatant (80 μ g) had antifungal activities but not
antibacterial activities. And MJ1-4 had higher antifungal activities than CH51.
When concentrated culture supernatant was treated with proteases, MJ1-4 and
CH51 lost activity partially by pepsin treatment but were not affected by other
enzymes. The results indicated that antimicrobial activities of these bacilli might
be derived from various compounds including bacteriocin(s). HA Lee, JY Park,
JY Lee, and CU Baek were supported by the 2nd stage Brain Korea 21
p
rogram
from the Ministry of Education and Human Resources Development, Republic
of Korea
Keywords: antimicrobial activities, B. amyloliquefaciens
I-35
The Effect of Poly-γ -Glutamic Acid (γ -PGA) on Intestinal
Absorption of Calcium and Comparison with
Caseinphosphopeptide
Jae-Chul CHOI
1
, Doo-Yeol CHOI
2
, Seung Ho LEE
3
, Sang-Yun LEE
3
,
Il-Han LEE
1
and Moon-Hee SUNG*
1,2
1
BioLeaders Corporation, Daejeon, Korea.
2
Dept. Bio &Nanochemistry,
K
ookmin
University, Seoul, Korea.
3
Vital Lab R&D Center, Vital Lab Co., Ltd., Seoul,
Korea.
*Corresponding author: smoonhee@bioleaders.co.kr
The bioavailability of calcium is currently one of the most important
topics in nutrition research and is correlated with gastrointestinal solubility.
Thus, to increase the solubility of calcium, we applied the poly-γ
-glutamic acid, an edible and biodegradable polymer from Chungkookjang
with repeated-chain structure of glutamate. And we investigated the effect
of γ -PGA on the soluble calcium uptake in the small intestine using
rat as the animal model system. The amount of soluble calcium uptake
in the rat was significantly high by adding γ -PGA(2,000 kDa)
Accordingly, our results strongly support the notion that γ -PGA has an
effect on calcium solubility, due to the formation of chelate of calcium
and γ -PGA, therefore the release of calcium sustained in the small
intestine. And we tried the increasing ability of calcium solubility was
compared with CPP(Caseinphosphopeptide) in vitro. These studied may
suggest more effective applications of calcium supply for the dietary
supplements. [This work was supported by the 'Seoul R&BD
Program(10580)’]
Keywords: poly-γ -glutamic acid, intestinal absorption of calcium,
caseinphosphopeptide
I-36
353
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Screening and Characterization of Saccharomyces cerevisiae
Isolated from Different Sources for the Development of
Direct-fed Microbials
Myeong Su SHIN*, Ae Ran JI and Kyeong Su KIM
Organic Bio Tech Co., Ltd. Jincheon, Chungbuk 365-861, Korea.
*Corresponding author: msshin21@hanmail.net
In order to develop probiotic yeast for livestocks, Saccharomyces
cerevisiae strains were isolated from commercial products and food
samples (brewer’s grain) and identified using API 20C AUX kit. The
strains were characterized by probiotic selection criteria such as tolerance
to acid and bile, and heat resistance. Most of them were resistant to 0.5%
bile salts and remained viable after 2 h at pH 2.0. Among the strains
tested, S-4 strain showed survival percentage

80% when exposed to
acidic and bile environment, respectively. None of the strains tested
survived or recovered after heat treatment in 50mM phosphate buffer at
60

for 1 h. The highest heat resistance was observed in S-4 strain
showing 3% of survival rate after heat treatment at 50

for 1 h. The
two strains of S. cerevisiae isolated from brewer’s grain showed higher
resistance to ethanol at 5%, 10%, and 15%(v/v).
Keywords: Saccharomyces cerevisiae, direct-fed microbials, livestocks
I-37
Optimizing Cultivation Process for Bioactive Compounds
Production of Cordyceps militaris Grown on Germinated
Soybean Using Metaolomics Approach
Jung Nam CHOI, Jiyoung KIM, Mi Yeon LEE, Ah Jin KIM,
Dong Ki PARK and Choong Hwan LEE*
Dept. of Bioscience and biotechnology, Konkuk University, Seoul 143-701,
K
orea.
*Corresponding author: chlee123@konkuk.ac.kr
Cordyceps militaris grown on germinated soybeans (CMGGS) might
be a promising efficacious source of novel bioactive compounds.
LC-ESI-MS based on metabolomics approach was used to investigated
the variation of secondary metabolites from week dependently cultivated
CMGGS. The PCA scores plots showed clear metabolites differences
between cultivation times and these metabolites were identified as
soyasaponins, daidzein, genistein, glycitein and their glycosides
b
y
comparing MS/MS spectral data with in-house MS/MS library. PCA scores
plots showed that major fermentation process and metabolite changes were
completed within 3 weeks after the inoculation with Cordyceps militaris.
During the cultivation, the antioxidant activity and total flavonoid contents
were showed a similar change with movement of PCA scores plot and
most highly indicated at 1 weeks cultivated. Also, discriminatory
compounds among the antioxidant activity were soyasaponins and
isoflavone aglycones according to PLS biplots. The information of this
study is valuable for the optimizing cultivation process for
b
ioactive
compound production of CMGGS using LC-ESI-MS based on
metabolomic study. In addition, several novel isoflavone methyl-
glucosides were detected and those structures were identified by NMR
spectroscopy.
Keywords: Cordyceps militaris, metabolomics, PCA anlaysis
I-38
Optimization of Fermentation Condition for Preparation of
Korean Traditional Rice Wine (Farmer Rice Wine)
Dae Hyoung LEE*
1
, Heui-Yun KANG
1
, Soonjae KIM
1
and Jong Soo LEE
2
1
1Gyeonggi-Do Agricultural Research and Extension Services,
H
wasung,
449-702, Korea.
2
Dept. of Genetic Engineering, Paichai University,
D
aejeon,
Korea.
*Corresponding author: leedh2@gg.go.kr
The objective of this study was to develop a Korean traditional rice
wine (farmer rice wine). Effects of fermentation temperature(10~30

),
addition of nuruks(1.0~3.0%) and yeast(0.5%~1.6%) on the quality of
Korean traditional rice wine were investigated. The maximum amount
if ethanol (18.4±0.5%) was produced when 2% nuruk and 0.8% yeast
were added to cooked rice and fermented at 20

for 10 days. The ethanol
contents ranged from 12.7 to 18.4%, while the residual sugar levels were
in the range of 1.62±0.4 to 13.45±0.5 mg/ml. In sensory evaluation,
traditional rice wine made from the addition of 2% nuruk and 0.8% yeast
showed the best overall acceptability. Temperature for the optimum
fermentation should be maintained in the range of 15

~25

. Optimal
fermentation conditions was addition of 1% nuruk and 0.5% yeast at 25

fermentation and 2% nuruk and 0.8% yeast at 20

fermentation.
Keywords: farmer rice wine, fermentation, nuruk, yeast
I-39
Extraction of Proanthocyanidin from Pine (Pinus densiflora)
Needles by Solvents and Subsequent Sub-Supercritical CO
2
Extractions
Sang-Hyeon LEE*
1
, Nam-Young KIM
1
, Min-Kyung JANG
1
, Ki Hwan YU
1
,
Hye Ji JANG
1
, Yong Soo PARK
2
, Mihyang KIM
2
, Sung Gu KIM
3
and
Byung Hong YOO
3
1
Dept. of Pharmaceutical Engineering, Silla University, Busan 617-736,
K
orea.
2
Dept. of Food and Nutrition, Silla University, Busan 617-736, Korea.
3
Bioport
Korea Co., Busan, 619-912 Korea.
*Corresponding author: slee@silla.ac.kr
In this investigation, we investigate to establish the most effective method
for proanthocyanidin extraction from pine needle. Pine needle extracts
p
repared
by using ethanol and hexane revealed high proantocyanidin contents out of
total polyphenolic compounds. To purify proanthocyanidin further from ethanol
and hexane extracts, these extracts were subsequently extracted
b
y
sub-supercritical CO
2
. Analyses of total polyphenolic compounds and
proanthocyanidin in each extracts indicated that ethanol-sub-supercritical (ESS)
extract contained higher proanthocyanidin content than finely ground
powder-sub-supercritical (FGPSS) extract which was directly extracted from
finely ground pine needles by sub-supercritical CO
2
. Furthermore,
sub-supercritical (SS) extract also revealed higher proanthocyanidin content
than FGPSS. Antioxidative activities of both ESS and FGPSS extracts showed
higher antioxidative activities than those of SS extract. These results suggest
that combinational extraction methods using solvent and sub-supercritical CO
2
are more effective to achieve extracts that contained high concentration of
proanthicyanidin. Acknowledgement: This work was support by Human
Resource Training Project for Regional Innovation, Ministry of Knowledge
Economy, Korea.
Keywords: proanthocyanidin, antioxidative activity, pine needle
I-40
354
www.kormb.or.k
r

Heterologous Production of Pediocin PA-1 in Lactobacillus
reuteri
Gi-Seong MOON* and Ji-Eun EOM
Div. of Food and Biotechnology, Chungju National University,
J
eungpyeong,
Chungbuk 368-701, Korea.
*Corresponding author: gsmoon@cjnu.ac.kr
Pediocin PA-1 is a representative class

a bacteriocin showing a strong
antilisterial activity and have been heterologously produced in several
bacterial strains. In this study, pediocin PA-1 structural and immunity
genes (pedAB) fused with the promoter and deduced signal sequence of
an α -amylase gene from a bifidobacterial strain were inserted in pLR5cat,
an Escherichia coli-lactobacilli shuttle vector. The recombinant DNA
pLR5cat_PSAB was transferred to Lactobacillus reuteri KCTC 3679 and
the transformant showed bacteriocin activity. The recombinant DNA was
minimized by removing a region of pLR5cat which is not essential for
the plasmid replication in lactobacilli. The recombinant L. reuteri KCTC
3679 transformed by the minimized pLR5cat(S)_PSAB also showed
bacteriocin activity. Molecular mass of secreted pediocin PA-1 from the
recombinant bacterium was same as that of native pediocin PA-1 from
Pediococcus acidilactici K10 on a sodium dodecylsulfate-polyacrylamide
gel electrophoresis (SDS-PAGE) gel. The recombinant L. reuteri KCTC
3679 effectively reduced the viable cell count of Listeria monocytogenes
by 3 log scale in a co-culture compared with a control where
L
is.
monocytogenes was inoculated alone.
Keywords: pediocin PA-1, Lactobacillus reuteri, heterologous production
I-41
Effect of Intestinal Bacteria by Enzymatic Hydrolysates of
Curdlan
Gwi Gun PARK* and Kyung Soo JEONG
Dept. of Food &Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
This report was performed to study on find out two group of
oligosaccharides (gentiobiose and laminaribiose) of which different Rf
value by controlled enzyme concentration and substrate concentration.
Aspergillus niger β -glucanase was produced with the optimum production
medium composed of yeast extract 5%, peptone 2%. At the optimal culture
condition, the production of crude enzyme reached the highest levels of
after 144hr cultivation at 35

, 120rpm. Curdlan was hydrolyzed into
gentiobiose by the crude β -glucansse. Gentiobiose were produced
quantitatively by activated carbon column chromatography. To investigate
of the effects of gentiobiose on the in vitro growth of Bifidobacterium
spp., various species were cultivated individually on the modified-MRS
medium contaning carbon source, such as gentiobiose, respectively.
Keywords: gentiobiose, laminaribiose
I-42
In vitro Antioxidant Activity and α -Glucosidase Inhibitory
Potential of By-product Produced by Grape Fermentation
Myung-Hee KIM, Kyoung-Soo HA, Dong-Soo KIM, Sung-Hoon JO,
Eun-Ji CHOI, Hae-Dong JANG and Young-In KWON*
Dept. of Food and Nutrition, Hannam University, Daejeon 305-811,
K
OREA.
*Corresponding author: youngk@hnu.kr
In the current study, we evaluated the inhibitory activity of
b
y-product
produced during fermentation of grape against α -amylase and α
-glucosidase. We also investigated phenolic-linked antioxidant activity,
oxygen radical absorbance capacity (ORAC) and sugar content with
fermentation time. Antioxidant activity showed similarly between grape
juice and fermented-grape juice and ethanol extracts of fermented-grape
skin had 3 times higher antioxidant activity than water extracts. α
-Glucosidase inhibitory activity increased moderately with fermentation
after 24 h. The water extracts of fermented-grape skin had higher α
-glucosidase and α -amylase inhibitory activities than that of ethanol
extracts. Sugar content of grape was significantly decreased
b
y
fermentation. These results indicate that fermentation of grape resulted
in mobilization of total phenolics which could be effectively designed
as complimentary therapies for postprandial hyperglycemia and oxidative
stress-induced diabetic complications. Due to by-product of fermented
grape had high antioxidant activity and carbohydrate hydrolyzing enzymes
inhibitory activity this approach also would have the potential to contribute
as a useful dietary strategy for management of postprandial hyperglycemia
and diabetic complications.
Keywords: antioxidant, fermented grape, α -glucosidase inhibitory
activity
I-43
Effects of Cooked and Uncooked Rice, and Gruel Rice on
the Preparation of Korean Traditional Rice Wine
Dae Hyoung LEE*
1
, Heui-Yun KANG
1
, Myeong whoon SEO
1
and
Jae Ho KIM
2
1
Gyeonggi-Do Agricultural Research and Extension Services, Hwasung, 449-702.
2
Korea Traditional Food Research Division, Korea Food Research Institute.
*Corresponding author: leedh2@gg.go.kr
This study was carried out to investigate fermentation characteristics
of cooked and uncooked rice, and gruel rice in the brewing of Korean
traditional rice wine. Changes of physicochemical properties and sensory
evaluation of traditional rice wine were investigated during fermentation
by using cooked and uncooked rice, and gruel rice. After 3 days
fermentation sugar content of the cooked rice wine and gruel rice wine
were about 11 brix, while uncooked rice wine was 6 brix. Total acid
contents of gruel and uncooked rice wine were also 0.25% and 0.20%,
respectively and cooked rice wine was 0.17%. Alcohol contents of
uncooked rice wine and gruel rice wine were significantly increased from
10.6% and 9.9% after 3 days fermentation to 17.1% and 17.2% after
7 days fermentation. Sensory evaluation revealed gruel rice wine was the
best in total acceptability with sweety and refreshing tastes.
Keywords: cook rice, uncooded rice, gruel rice, fermentation
I-44
355
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Antibacterial Activities of Glyceollins Extracts
Ae Ran LEE
1
, Mee Ryung LEE
1
, Kang Wook LEE
2
, Ji Yeon LEE
2
,
Ji Yeong PARK
2
, Hwang A LEE
2
, Chang Un BAEK
2
, Hyeon Deok JO
2
,
Joo Yeon KIM
2
and Jeong Hwan KIM*
1
1
Institute of Agriculture and Life Science, Gyeongsang national university,
J
inju
660-701.
2
Division of Applied Life Science (BK21 program), Graduate School
Gyeongsang National University, Jinju 660-701.
*Corresponding author: jeonghkm@gsnu.ac.kr
The production of glyceollins, a known phytoalexins, is induced upon
fungal infection soybean. In this study, we investigated the antibacterial
activities of glyceollins using three types of soybean sample; control
(soybean soaked in the water), soybean germinated, soybean germinated
and fungal-infected. Paper disc methed was used to examing anti microbial
activities of soybean samples. The results showed that soybean sample
with fungal infection had microbial activities and inhibited B.
s
ubtilis,
B. cereus, B. licheniformis, B. circulans, Salmonella typhimurium, and
E. coli 0157 : H7. B. cereus was inhibited the most. JY Lee, HA Lee,
JY Park and CU Baek were supported by the 2nd stage Brain Korea
21 program from the Ministry of Education and Human Resources
Development, Republic of Korea.
Keywords: glyceollins, phytoalexins, antibacterial activities
I-45
Molecular Cloning of the Genes for GDP-Mannose 4,
6-Dehydratase and GDP-L-Fucose Synthetase from
Bacteroides thetaiotaomicron
Mi-Hee JANG, Jong-Gil YOO and Myoung-Dong KIM*
School of Bioscience and Biotechnology, Kangwon National University,
Chuncheon 200-701, Korea.
*Corresponding author: mdkim@kangwon.ac.kr
Genes encoding GDP-
L
-fucose synthetase and GDP-mannose 4,
6-dehydratase which are crucial enzymes for biosynthesis of GDP-
L
-fucose
were cloned from Bacteroides thetaiotaomicron and overexpressed in
recombinant Escherichia coli using IPTG(isopropyl-1-thio-β -D-
galactopyranoside)-inducible expression vector. Deduced amino acid
sequences of GDP-
L
-fucose synthetase from B. thetaiotaomicron showed
46% and 28% homology with those of E. coli and Helicobacter
p
ylori,
respectively. In the case of B. thetaiotaomicron GDP-mannose 4,
6-dehydratase, it showed highest similarity with that of E. coli. Maximum
levels of soluble expression for those two proteins in recombinant E. coli
were achieved at 30

and 0.1 mM concentration of IPTG. Functional
expression of two key enzymes was validated by measuring GDP-
L
-fucose
content of recombinant E. coli strain.
Keywords: GDP-L-fucose, GDP-L-fucose synthetase, GDP-mannose 4,
6-dehydratase
I-46
Evaluation of Probiotic Characteristics and Safety of Lactic
Acid Bacteria Isolated from Korean Infant Feces and Kimchi
Jin LEE, Hyun Sun YUN, Gi Hoon JEUN, Min Young YOO,
Seul LEE, Jung Hwa KANG and Sae Hun KIM*
Division of Food Bioscience and Technology, Korea University, Seoul 136-701,
Korea.
*Corresponding author: saehkim@korea.ac.kr
Lactic acid bacteria (LAB) inhabit intestinal tracts of humans and
animals having beneficial physiologic effects. LAB usually regarded as
safe, but some reported that it causes rare infections or haves antibiotic
resistances. The aim of this study is evaluation of probiotic characteristics
and safety. 200 strains LAB isolated from Korean infant feces and kimchi
was investigated. The strains were tested their common probiotic abilities
such as acid tolerance (pH 2.5 / 0.3% oxgall), and adhesion to human
intestinal epithelial cells (HT-29 cell). Among the strains, some are mainly
focused on functional aspects. Antimicrobial activities and cholesterol
assimilation were examined, and C. elegans killing assay were done.
Hemolytic type, gelatin hydration, and degradation of urea were also
assessed to certifying their safety in the host. Of tested strains, five LAB
could assimilate the cholesterol and three strains could prolong the life
time in C. elegans model having acid/bile tolerance and adhesion ability.
And, most LAB could inhibit pathogenic bacteria, and their safety were
proven. The strains were identified as Lb. fermentum, Lb. gasseri, and
Lb. plantarum strains by 16S rDNA sequencing. These selected strains
could have potentials to use of food application or feed additives.
Keywords: probiotics, lactic acid bacteria, safety
I-47
Isolation of Bacteria for Sea Mustard Fermentation and its
Culture Characteristics
Ji-Hun CHOI
1
, Xiao-Tian QUAN
1
, Yun-Jung LEE
1
, Sang-Buem CHO
1
,
Dong-Woon KIM
2
and Soo-Ki KIM
1
*
1
Department of Animal Science and Technology, Konkuk University, Seoul
143-701, Korea.
2
National institute of Animal Science, RDA
*Corresponding author: sookikim@konkuk.ac.kr
Various bacteria were isolated from naturally fermented sea mustard
and its culture characteristics were investigated in enrichment media
containing sea mustard powder. In the first experiment, 20% of sea mustard
powder was added in YM, LB, MRS and PD broths. For the natural
fermentation, each broth was divided into two groups: one was sterilized
by autoclaving and the other was not. Each broth was incubated for 50
days at room temperature without any inoculation. By the characteristics
of morphology and antibiotic resistance, various colonies were isolated
from the cultures. Herbaspirillum sp, and Chryseobacterium sp. were
isolated on YM culture, B.subtilis, Enterococcus faecium, and
Enterococcus faecium were isolated on LB, MRS, and PD cultures,
respectively. In the second experiment, B. subtilis SK877 and
Lactobacillus brevis SK1304 were inoculated on LB and MRS media
containing 1~15% sea mustard powder, respectively. B. subtilis SK877
showed a good growth in both media whereas Lactobacillus brevis SK1304
did not. As a result, B. subtilis SK877 was more effective for the
fermentation of sea mustard. (This Study was supported by Technology
Development Program for Agriculture and Forestry, Ministry for
Agriculture, Forestry and Fisheries, Republic of Korea)
Keywords: sea mustard powder, fermentation, culture characteristics
I-48
356
www.kormb.or.k
r

Nanoscale Observation of Antibacterial Effects of Green Tea
Polyphenol (-)-Epigallocatechin-3-Gallate at Sub-Minimum
Inhibitory Concentrations (sub-MICs) on Escherichia coli
O157:H7
Yidan CUI, Jeesun LIM, Seong-Won NAM and Sungsu PARK*
Department of Chemistry and Nano Sciences, Ewha Womans University, Seoul
120-750, Korea.
*Corresponding author: janechoi85@hanmail.net
Epigallocatechin-3-gallate (EGCG) is known to have antibacterial
properties, but the exact mechanism underlying how EGCG exerts the
properties has not been elucidated yet. Herein, we studied the effects of
EGCG at sub-MICs (10 to 60mg/mL ) on the morphology of Escherichia
coli O157:H7 using atomic force microscopy (AFM). Our results showed
that EGCG disturbed bacterial membrane and induced the formation of
vesicles. The extent of vesicle formation is clearly dependent on the
concentration of EGCG and its duration time in the culture. The lysis
of cell membrane was not observed even at the highest concentration-60
mg/mL and 18 hr-incubation, confirming that EGCG at sub-MICs is
sub-lethal. Our results directly show visual and quantitative structural
changes of bacterial membranes induced by EGCG.
I-49
An Antistaphylococcal Bacteriocin Enterocin MK3 from
Enterococcus faecium MK3
Gi-Seong MOON*, Cheon-Ho SONG and Kyung-Sook KIM
Div. of Food and Biotechnology, Chungju National University,
J
eungpyeong,
Chungbuk 368-701, Korea.
*Corresponding author: gsmoon@cjnu.ac.kr
Staphylococcus aureus is very important pathogen due to its antibiotic
resistance such as MRSA (methicillin-resistant S. aureus) and VRSA
(vancomycin-resistant S. aureus). Development of next generation
antibiotics for treatment of the bacteria are urgently needed. In this respect,
bacteriocins inhibiting S. aureus could also be alternative
antistaphylococcal agents. Enterocin MK3 from Enterococcus faecium
MK3 showed bacteriocin activities against some Gram-positive
b
acteria
including S. aureus and Listeria monocytogenes. Inhibition pattern of the
bacteriocin appeared to be bactericidal. The molecular mass of enterocin
MK3 was estimated to be approximately 6.5 kDa on a SDS-PAGE (sodium
dodecyl sulfate-polyacrylamide gel electrophoresis) gel and the
b
acteriocin
band was subjected to MALDI-TOF (Matrix-Assisted Laser Desorption/
Ionization-Time Of Flight) analysis although we could not get any
information from it. At the moment, the genes for the enterocin MK3
production are under investigation.
Keywords: enterocin MK3, Enterococcus faecium, Staphylococcus aureus
I-50
Lactobacillus curvatus HY7601 Reduces Cholesterol Level and
Body Weight in Mice
Do-Young PARK*, Young-Tae AHN, Hoyong LEE, Jung-Hee LEE and
Chul-Sung HUH
R &D Center, Korea Yakult Co., Ltd., 418-12, Korea.
*Corresponding author: ehdud@re.yakult.co.kr
Several studies have reported that ingestion of specific strains of
lactobacilli could reduce serum cholesterol levels in humans. However,
the cellular mechanism producing this reduction remains unclear. Thus,
we suggest that L. curvatus HY7601 mediates this effect by affecting
cellular sterol homeostasis. As a critical regulator of sterol homeostasis,
under cholesterol-depleted conditions, sterol-regulatory element
b
inding
protein-2 (SREBP-2) is subjected to proteolysis and bind to sterol response
element (SRE) of genes involved in cholesterol synthesis and uptake,
which up-regulates expression of those genes. When HepG2 cells were
transiently transfected with SRE-containing reporter plasmids, the reporter
expression increased in response to L. curvatus HY7601. In HepG2 cells
treated with L. curvatus HY7601 the mature form of SREBP-2 increased
and SRE-regulated genes such as HMG-CoA reductase and surface LDL
receptor expression also increased. In vivo, in mice fed a L. curvatus
HY7601 with high-fat diets, serum cholesterol levels was remarkably low
compared to those of mice fed only high-fat diets. In addition, body weight
of mice fed L. curvatus HY7601 was also considerably reduced. We
conclude that L. curvatus HY7601 has dramatic effect on both cholesterol
level and obesity.
Keywords: L. curvatus HY7601, cholesterol, anti-obesity
I-51
Application of Poly-γ -Glutamic Acid Improves Dermatitis
in NC/Nga Mice, a Model of Human Atopic Dermatitis
Tae young LEE
1
, Jai-Chul CHOI
2
, Moon-Hee SUNG
2,3
and
Haryoung POO*
1
1
Infectious Disease Research Center, Korea Research Institute of Bioscience an
d

Biotechnology, Daejeon, Korea.
2
Bioleaders Cooperation, Daejeon, Korea.
3
D
ept.
Bio &Nanochemistry, Kookmin University, Seoul, Korea.
*Corresponding author: 9403045@hanmail.net
Atopic dermatitis (AD) is a chronic inflammatory skin disease
characterized by pruritic and eczematous lesions, along with increased
total IgE level in plasma, inflammatory cell infiltration, and the expression
of T helper 2 (Th2) cytokines. NC/Nga mice kept in conventional
conditions are known to develop skin lesion resembling human AD. Poly-
γ -glutamic acid (γ -PGA) is an edible biomaterial naturally synthesized
by Bacillus subtilis (chungookjang). Previously, we reported that γ -PGA
induced TLR4-dependent activation of innate immunity, such as the
production of IL-12 to shift the immune response from type Th2 to Th1.
To examine the effect of γ -PGA on NC/Nga, the γ -PGA was applied
once a day for 7 weeks, starting at 14 weeks old on to the shaved skin.
γ -PGA remarkably changed the immune response from Th2 and Th1
as determined from cytokine level. The serum IgE and IgG1 level was
decreased and the expression of IgG2a was upregulated. Also, the splenic
level of IL-10 was downregulated, whereas that of IFN-γ and IL-12
was increased. Also, histological analysis of the skin revealed that
application of γ -PGA significantly inhibited the thickening of epidermis
compared to control mice. These results suggest that γ -PGA is effective
for immunotherapy agents for the treatment of AD.
Keywords: atopic dermatitis (AD), poly-γ -glutamic acid
I-52
357
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Enhancement of Muscle and Body Weight by Orally
Administrated Lactobacillus casei Expressing Surface
Displayed Myostatin
Seung Pyo HONG
1
, Jong-Tak KIM
3
, Haryoung POO
4
, Chul-Joong KIM
1,5
and Moon-Hee SUNG*
1,2
1
BioLeaders Corporation, Daejeon, Korea.
2
Dept. of Bio &Nanochemistry,
Kookmin University, Seoul, Korea.
3
Greentherapy Corporation, Gyeonggi,
K
orea.
4
Korea Research Institute of Bioscience and Biotechnology, Daejeon,
K
orea.
5
College of Veterinary Medicine, Chungnam National University, Daejeon,
K
orea.
*Corresponding author: jongbok@bioleaders.co.kr
Myostatin, a member of the TGF-superfamily, is a potent negative regulator
of skeletal muscle growth. The repression of myostatin has been showed a
marked increase in muscle mass and body weight. The objective of this study
is to examine the effect of myostatin inhibition on skeletal muscle mass in
mice and chicken. In this study, we create the Lactobacilli casei, expressing
the mature domain of chicken myostatin (CMyom) on its surface. Surface
expression of CMyom protein on L. casei was confirmed by immunoblot and
FACScan analysis. Oral inoculation of B6SJL mice with L. casei expressing
CMyom induced antibody responses specific for CMyom and increased
b
ody
weight compared to the control group. In chicken, oral inoculation of L. casei
expressing the CMyom resulted in increased level of serum IgG antibodies
against Cmyom as well as an increase in body weight compared to those of
the control group at 5 weeks post feeding. The results of this study indicate
that myostatin expressing L. casei can induce an immune response resulting
in myostatin blockage, and this can be a potential means to improve skeletal
muscle growth and body weight of chickens. [This work was supported
b
y
DAEDEUK INNOPOLIS from the Ministry of Science and Technology of
Korea]
Keywords: Lactobacillus casei, myostatin
I-53
Isolation and Characterization of ACE Inhibitor Producing
Bacteria from Traditional Fermented Foods
Chun pyo JEON
1
, Jung Bok LEE
2
, Seong Yeon CHOI
1
, Chung Sig CHOI
3
,
Oh Seuk LEE
4
and Gi Seok KWON*
1
1
School of Bioresource Sciences, Andong National University, Andong 760-749,
Korea.
2
Dept. Food &Nutrition Science, Kundong University, Andong 760-833,
Korea.
3
Bio Industry Institute, HansBio Co., Ltd. Andong 760-380, Korea.
4
D
ept.
of Food Fermentation Technology, Youngdong University, Youngdong 370-701,
Korea.
*Corresponding author: gskwon@andong.ac.kr
In this study, lactic acid bacteria with high ACE inhibitor producing
ability were isolated from kimchi and salted seafood. The strain HLJ59,
isolated from salted shrimp showed the highest ACE inhibitor producing
ability of 98.83% in MRS broth. Optimum growth temperature of HLJ59
was 4

and acid treatment at pH 3.0 for 1.5 hr, in all the selected lactic
acid bacteria, the intial viable cells of about 9.9×10
8
CFU/

decreased
to about 3.11×10
4
CFU/

HLJ59. The NaCl concertration of 3.0% in
MRS broth little better than inhibit the growth of HLJ59, but almost
decreased in the concentration of 5%, 7% and 10%. The bile extract
concentration of 0.3%, 0.5% and 1.0% in MRS broth did not inhibit the
growth of HLJ59. These studies suggest that yoghurt fermented with
appropriately selected lactic acid bacteria may have a antihypertension
effect.
Keywords: ACE inhibitor, lactic acid bacteria, fermented food
I-54
Enhancement of Antioxidant Activity of Codonopsis lanceolata
By Lactic acid Fermentation
Seung Seop Kim, Ji Hye Ha, Hyang Suk Jeong, Myoung Hoon Jeong,
Sung Ho Oh, Ling Jin, Jae Gun Han and Hyeon Yong Lee
Dept. of Biomaterials & Engineering, College of Bioscience & Biotechnology,
Kangwon National University, Chuncheon 200-701, Korea
*Corresponding author: geniusgene@kangwon.ac.kr
Codonopsis lanceolata has mainly been used as a food that act much
population due to its flavors. Therefore in this studies, Codonopsis
lanceolata was fermented by Leuconostoc mesenteroides with MRS media
to improve its efficacy. There was dramatic increase of phenolic contents
from fermentation of Codonopsis lanceolata and SOD scavenging activity
was also increased. The total phenol contents of fermented Codonopsis
lanceolata were as 27.9 μ g/

, compared to 5.4 μ g/

fresh Codonopsis
lanceolata. Fermented Codonopsis lanceolata effectively inhibition
SOD-like activity as 21.56%, which was much higher than 16.5% of fresh
Codonopsis lanceolata. From the result, fermentation of Codonopsis
lanceolata can be enhance its biological activity by increasing the amounts
of active components as well new substances from Codonopsis lanceolata.
Keywords: Codonopsis lanceolata, antioxidant, total phenol, SOD
I-55
Bacilus subtilis Spore Display System
Junehyung KIM*
2
and Byung-Gee KIM
3
1
Department of Chemical Engineering, Dong-A University, Busan, Korea.
2
동아
대학교.
3
School of Chemical and Biological Engineering, Seoul National
University, Seoul, Korea.
*Corresponding author: june0302@dau.ac.kr
Bacterial surface display finds its important biotechnological application
in the fields of screening tools of evolved enzyme, bioremediation, whole
cell bioconversion and tool for live vaccine production [1]. For the
functional bacterial surface display of active enzyme of multimeric form,
which is generally impossible due to molecular assembly of the monomer
subunit subsequent to the secretion of displayed target protein outside
the cell, a new surface display system based on Bacillus subtilis spore
was developed. Using cotE and cotG motif, β -galactosidase, which is
active only in tetrameric form, was functionally displayed on the surface
of Bacillus subtilis spore. The surface localization of β -galactosidase was
verified by enzymatic assay of purified spore, protease accessibility test
and FACS analysis of spore expressing β -galactosidase. While
B
acillus
subtilis spore wall integrity was slightly affected by the incorporation o
f

CotE-LacZ fusion protein, it was not affected by the incorporation of
CotG-lacZ fusion. Other enzymes such as streptavidin, lipase (lipB) of
Bacillus subtilis and GFP
UV
were also successfully displayed using cotE
and cotG motifs, and its possible application in various
b
iotechnological
fields will be discussed [2, 3].
Keywords: surface display, Bacillus subtilis, spore
I-56
358
www.kormb.or.k
r

Improvement of Crude Rice Wine with Purple Sweet Potato
which Follows in Addition Time
Heui-Yun KANG*, Daehyoung LEE, Yongseon LEE, Myeongwhoon SEO,
Soonjae KIM and Kyeong-yeol PARK
Division of Horticultrue Resarch, Gyeonggido Agricultural Resarch &Extexnsion
services, Hwasung, 449-702, Korea.
*Corresponding author: yeastlove@gg.go.kr
We improved crude rice wine with purple sweet potato that the alcohol
was produced 16.7±0.2% (increased 17% alcohol yield) under condition
steamed rice 70%, steamed purple sweet potato 30%, improved nuruk
(Hankook Enzyme) for rare rice 0.2%, yeast (Saccharomyces cerevisiae)
0.08%, at 25

during 10 days. Specially, antioxidant activity and color
for crude rice wine with purple sweet potato were the highest and stabilized
in pH 3~5. As red natural pigment, the cyanidin 3-glucoside(C3-G) is
the main anthocyanin pigments of purple sweet potato. It will be getting
the added value with purple sweet potato that is higher controlling than
wild grape cake as natural color material. The extractions of the pigment
it will be able to accomplish easily even from fermentation. The crude
rice wine with purple sweet potato will be desirable to do from the
agricultural association or brewery.
Keywords: anthocyanin, crude rice wine (makgeolli), purple sweet potato
I-57
Effect of Pulsed Electric Field on Ethanol Fermentation and
Chemical Component Variation in a Wine-making Culture
Doo Hyun PARK*, He-Ryeon MIN, Bo Young JEON, In Jin SONG and
Jun Cheol KIM
Dept. of Biological Engineering, Seokyeong Univ. Seoul 136-704, Korea.
*Corresponding author: baakdoo@skuniv.ac.kr
The aim of this research was to estimate the effect of pulsed electric
field (PEF) on yeast metabolism and chemical component variation of
a wine-making culture. Electric polarity of working electrode and counter
electrode was periodically switched at the intervals of 30s. Electric current
generated by anodic and cathodic reaction of working electrode was
reached to +30 mA and-12 mA, respectively, and induced the PEF in
the wine-making culture. The yeast growth, ethanol production, and malate
consumption in the initial cultivation time were more activated in the
PEF than the conventional condition (CC) and proportional to the potential
intensity of PEF. Polyphenol, total phenolic contents (TPC), and total
flavonols were gradually decreased in all cultivation conditions during
cultivation for 2 weeks but antioxidation activity was not. TF was
significantly lower in 3 and 4 volt of PEF than CC and 2 volt of PEF;
however, the polyphenol, TPC, anthocyanin, and antioxidation activity
were a little influenced by the PEF. The sensuously estimated aroma,
flavor, and taste of the test wines produced in the CC and 2 volt of
PEF was significantly different from those produced in 3 and 4 volt of
PEF.
Keywords: wine-making culture, pulsed electric field, Saccharomyces
bayanus
I-58
Isolation of β -Glucosidase Gene from Bacillus licheniformis
KJ-3 and its Expression in E. coli
Hyeon Sook PARK, Byung Wook YANG and Young Tae HAHM*
Dept. of Biotechnology, Chung-Ang University, Anseong 456-756, Korea.
*Corresponding author: ythahm@cau.ac.kr
β -glucosidase (EC 3.2.1.21) catalyzes the hydrolysis on β (1

4)
linkage of disaccharides, alkyl-and aryl-glycosides. In this study, the β
-glucosidase gene of Bacillus licheniformis KJ-3 was isolated and
expressed heterologously in E. coli. The expressed proteins were also
characterized. According to the higher β -glucosidase activity,
B
.
licheniformis KJ-3 was selected out of the isolated Bacillus spp. obtained
from traditionally fermented soybean paste, Cheonggukang. The genomic
DNA of B. licheniformis KJ-3 was extracted and amplified using
oligonucleotide primers synthesized base on β -glucosidase gene
sequences of Bacillus spp. in NCBI. 1.4 kb of the PCR-amplified DAN
fragment was similar to bglH gene of B. licheniformis ATCC 14580 with
99% homology, which belong to glycoside hydrolase family 1. The
isolated bgl gene was subcloned in pET-25b(+) expression vector, which
contains T7 promoter. For heterologous expression, pET-25b(+)
containing bgl gene was transformed into E. coli BL21(DE3) and induced
with IPTG. Heterologously expressed proteins are being analyzed in
progress.
Keywords: β -glucosidase, gene expression, Bacillus licheniformis
I-59
Development of a White Wine with Campbell Early Grapes
Oh Seuk LEE*, Soyoung PARK, Wongook JUNG and Chul YOOK
Dept. of Wine &Fermented Food, Youngdong University, Chungbuk, 370-701,
Korea.
*Corresponding author: los25@hanmail.net
The freezing-press method applied from Ice wine's method that a
phenomenon of concentrated sugar by freeze concentration. In this
investigation showed changes of physicochemical properties during
fermentation between wine A(produced by freezing-pressure method) and
wine B(produced by chaptalization method that adding sugar at early stage
of fermentation). As a result the wine A has 24brix sugar contents, the
wine B has 26brix sugar contents. The wine A contains 13% alcohols,
the wine B contains 12% alcohols. The wine A contains 1.1% total acidity
as tartarate, the wine B contains 1.15% total acidity as tartarate. The wine
A showed 81.82 whiteness, 13.5 redness, 20.66 yellowness. The wine
B showed 33.92 whiteness, 65.64 redness, 30.80 yellowness. The wine
A has 0.46mg/mL polyphenolic compound, the wine B has 1.15mg/mL
polyphenolic compound. The wine B has much more polyphenolic
compound. The wine A produced closer white wine has high sugar
contents(without adding sugar) and polyphenolic compound and high
alcohol content, much flavor and little strong sour taste, showed light
red color(because fermented without skin), with freezing-pressure method.
Keywords: white wine, campbell early, freeze concentration
I-60
359
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Screening of Antiviral Activity from Natural Plants against
Feline Calicivirus : a Surrogate Norovirus
Kyoung-lan KIM and Myung-Suk LEE*
Department of Microbiology, Pukyong National University, Busan 608-737,
Korea.
*Corresponding author: leems@pknu.ac.kr
Noroviruse (NV), which is caused gastroenteritis illness in people, is
one of important food-borne pathogens of NV over world-wide. In an
effort to discover an antiviral against NV, we investigated antiviral effect
of various methanol extracts from natural plants including spices, herb
teas and medical herbs against feline calicivius (FCV) as a surrogate for
NV. The antiviral activity of extract was determined by plaque reduction
assay and virucidal effects performed the tissue culture infectious dose
50 (TCID
50
) against FCV. Among the eight extracts, green tea (Camellia
sinensis L.) exhibited strong antiviral activity against FCV with 50%
effective concentration(EC
50
) of 133

/mL and cytotoxicity concentration
(CC
50
) of 2470

/mL, its selectivity index(SI) was 18.57. And 1hr
treatment of the green tea extract in the presence of 3.13 mg/mL, FCV
was completely inactivated. The antiviral effect of Camellia sinensis L.
extract against FCV was determined to be does-and time-dependent. But
the others were weak antiviral activity against FCV. The results obtained
in this study suggest that the Camellia sinensis L. extract may be effective
in the prevention of food-borne disease cause by NV.
Keywords: norovirus, feline calicivirus, antiviral activity
I-61
Purification and Some Properties of β -Glucanase from
Aspergillus niger
Gwi Gun PARK*
2
and Su Yeon KIM
2
1
Dept. of Food &Bioengineering, Kyungwon University.
2
Dept. of Food
&Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
This report was performed to study on purification and structural
analysis of gentiobiose from the enzymatic hydrolysate of gums
b
y
Aspergillus niger. Aspergillus niger find out the effect of metabolism
activity of microorganisms of gentiobiose. Aspergillus niger β -glucanase
was produced with the optimum production medium composed of yeast
extract 5%, peptone 2%. At the optimal culture condition, the production
of crude enzyme reached the highest levels of after 144hr cultivation at
35

, 120rpm. Aspergillus niger β -glucanase was purified
b
y
DEAE-sephadex ion exchange column chromatography. The final
preparation thus obtained showed a single band on SDS-PAGE. The
purified β -glucanase exhibited maximum activity at pH5 and 60

.
Enzyme activity of the purified β -glucanase were affect condiderably
by treatment with Hg2+ and Mn2+.
Keywords: β -glucanase, Aspergillus niger
I-62
Changes in Bacterial Count and Enzyme Activity during
Fermentation of Kochujang Prepared with Wheat Flour
Song Yi HAN, Min Hwa KIM, Jin Bo JEONG and Yong Suk KIM*
Department of Food Science &Technology, Chonbuk National University,
Jeonbuk 561-756, Korea.
*Corresponding author: kimys08@chonbuk.ac.kr
Changes in bacterial count and enzyme activity of Kochujang prepared
with various wheat flours were investigated for 10 week at 25

. Glutinous
rice (GK), domestic wheat flour (DWK), and imported wheat flour (IWK)
were used as different starch sources. Aerobic bacteria counts of GK,
DWK, and IWK maintained at the level of 10
7
, 10
5
, 10
5
CFU/g during
fermentation, respectively. Acidic and neutral protease activities of GK
increased until 8 week of fermentation, decreased after 10 weeks. Those
of DWK and IWK were continually increased to 166.74~215.26

/g,
173.41~210.07

/g. α -Amylase activity was gradually decreased to
1.14~0.76 unit/g, 2.39~1.42 unit/g, 2.29~1.36 unit/g during fermentation.
After 10 weeks, that of DWK showed the higher 2.39 unit/g than those
of GK (1.14 unit/g) and IWK (2.29 unit/g). β -Amylase activity of all
treatments was rapidly decreased during fermentation.
I-63
Rapid and Simultaneous Detection of Food Microbial
Pathogen using DNA Chip
Jin Yong LEE, Ee Taek HWANG and Man Bock GU*
College of Life Sciences and Biotechnology, Korea University, Seoul, Republic
of Korea.
*Corresponding author: mbgu@korea.ac.kr
One of the most major public health problem is caused by food-borne
pathogen illness. Recently, method for the rapid and simultaneous
detection of food-borne pathogens is needed by the food industry, food
safety agencies and bioterrorism prevention organizations. And patterns
of food consumption have been remarkably changed and demands for
the safer foods are greatly increased. It has been reported that seventy
percents of food-borne diseases are caused by pathogenic microorganisms.
To prevent people from food-borne diseases, the pathogens should
b
e
detected from foods before the contaminated foods are distributed to the
consumers. In this study, we will develop a DNA chip to detect food-borne
pathogens rapidly and simultaneously. We selected Bacillus cereus 1013,
Staphylococcus aureus 1621, Samonella enteritica 2514 as food-borne
pathogens. For three microbial pathogens, we construct random DNA
library. Using random priming method, DNA chip will be also established
and optimized to enhance the sensitivity in detecting those
microorganisms. We are expecting that development of a rapid and
simultaneous detection method for hazardous microorganisms will enhance
the public health in Korea by reducing the occurrence of food-borne
diseases.
Keywords: DNA chip, foodborne pathogen, random library
I-64
360
www.kormb.or.k
r

Enhanced Productivity of Barley α -Amylase Isozyme 2 in
E. coli via Replacing the 42
nd
Alanine with Proline
Seung-Ho CHOI
1
, Myoung-Uoon JANG
1
, Jung-Mi PARK
1
, So-Ra LEE
1
,
Hong-Gyun LEE
1
, Birte SVENSSON
2
and Tae-Jip KIM*
1
1
Department of Food Science and Technology, Chungbuk National University,
Cheongju, 361-763, Korea.
2
Enzyme and Protein chemistry, Technical university
of Denmark, Copenhagen, Denmark.
*Corresponding author: tjkim@cbnu.ac.kr
Although barley α -amylase isozyme 1 (AMY1) and 2 (AMY2) share
up to 80% sequence identity and almost identical three-dimensional
structure, their enzymatic properties differ remarkably. To date, a number
of expression vectors and hosts have been examined to produce it in E.coli.
As a result, high amount of recombinant AMYs with C-terminal
six-histidines were successfully expressed under the control of inducible
T7 RNA polymerase promoter. However, AMY2 has been poorly
expressed under the inducible expression system in E. coli compared to
AMY1. In this study, the A42P mutant of AMY2 was successfully
constructed by site-directed mutagenesis. The 42
nd
Alanine residue is
located within the β -α -loop 2 of the catalytic (β/α )
8
-barrel domain.
Expression levels of AMY2 mutant A42P and wild-type AMY2 were
compared under the control of IPTG-inducible T7 RNA polymerase
promoter at 15

. As a result, about 10-times higher amount of
recombinant AMY2 A42P was obtained and simply purified via Ni-NTA
chromatography. Finally, Calcium-dependency and hydrolysis patterns on
various starch-derivatives of A42P mutant and wild-type AMY2 were
comparatively examined with each other.
Keywords: barley α -amylase, site-directed mutagenesis, expression level
I-65
Effect of γ -Aminobutyric Acid (GABA) Fermented from
Mono Sodium Glutamate (MSG) on Melatonin Secretion from
Mice
Seung Seop Kim, Ji Hye Ha, Hyang Suk Jeong, Myoung Hoon Jeong,
Sung Ho Oh, Ling Jin, Jae Gun Han and Hyeon Yong Lee
Dept. of Biomaterials &Engineering, College of Bioscience &Biotechnology,
Kangwon National University, Chuncheon 200-701, Korea.
*Corresponding author: geniusgene@kangwon.ac.kr
This study was performed to investigate the effects of GABA on
melatonin secretion, which is a hormone secreted from pineal grand of
brain. For the first time, GABA was produced by fermentation of MSG
whose concentration is 10%(w/v). GABA sample with the different
concentration (60

/

, 120

/

). The melatonin secretion was increased
as the mean of dosage of GABA in the mice. The highest melatonin
secretion (2.523 pg/

) observed in feeding 120

/

of GABA,
compared to the control(2.523 pg/

). It can tell that GABA can play
a role in enhancing the secretion of melatonin, which can be used as
a functional food for anti insomnia.
Keywords: γ -aminobutyric acid(GABA), melatonin, insomnia
I-66
Studies on Extraction and Antioxidant Properties of Hizikia
f
usi
f
ormis
Min Jeong SEO
1
, Min Jeong KIM
1
, Hye Hyeon LEE
1
, Woo Hong JOO
2
and Yong Kee JEONG*
1
1
Department of Medical Bioscience, Dong-A University, Busan 604-714,
K
orea.
2
Department of Biology, Changwon National University, Changwon, 641-773,
Korea.
*Corresponding author: ykj9912@dau.ac.kr
The antioxidative activities such as 1, 1-diphenyl-2-picrylhydrazyl
(DPPH) radical scavenging effect and SOD-like activity of two n-hexane
fractions (H1, H2), three ethyl acetate fractions (F1, F2, F3) and three
n-buthanol fractions (S1, S2, S3) obtained by open-column
chromatography of 80% ethanol extract of Hizikia fusiformis were
investigated. DPPH radical scavenging effect of each fractions were
increased in a dose-dependent manner, until the concentrations of 0.01~1
mg/ml with increasing of concentration, and H1 fraction showed highest
DPPH radical scavenging effect among prepared fractions. SOD-like
activities were also increased in a dose-dependent manner, until the
concentrations of 0.01~1 mg/ml with increasing of concentration.
However, the highest SOD-like activity showed at S1 fraction unlike
DPPH radical scavenging effect. These data suggest that H1 and S1
fractions have antioxidative compounds with different antioxidative
properties.
Keywords: SOD-like activity, hizikia fusiformis, DPPH radical
scavenging
I-67
Mixed Culture of Bacillus sp. Strains and Lactic Acid Bacteria
on the Fermentative Characteristics of Cheonggukjang
Mi Kyoung SHIN
1
, Yong Geun HA
1
, Jin Mi YOON
1
, Byoung Soo KIM
2
,
Young-Bae KIM
3
, Won Kap YUN
4
and Tae-Wan KIM*
1
1
Dept. of food science &Biotechnology Andong National University,
K
orea.
2
Gyeongbuk Institute for Bio Industry, Andong 760-380, Korea.
3
Nikkyuhbio,
Andong, 760-863, Korea.
4
Dept. of Marin Bio-Industry, Uljin, Korea.
*Corresponding author: silver0941@nate.com
Cheonggukjang is highly digestible, fibrinolytic activities, antimutation
and contributing important nutrients including calcium and vitamin A,
B, as well as functional properties, such as laxative effect and anticancer
properties. cheonggukjang was fermented by the action of lactic acid
bacteria and its biologically active substances can have functional effect
on the body. The aim of this work was to investigate the microbiological
and hysicochemical properties of the cheonggukjang prepared by mixed
culture of Bacillus sp. strains (MB2, MBL2, and KSW-3) and lactic acid
bacteria (MA2, MA4, MSE-1 and MKGS). Cheonggukjang was fermented
with Bacillus sp. strains (MB2, MBL2, and KSW-3) and lactic acid
bacteria (MA2, MA4, MSE-1 and MKGS) at 37

for 48 hours. After
addition of 0.5% skim milk soybean was fermented by single or mixed
culture. Cell colony count, slime material and protease activity of the
fermented cheonggukjang were measured and compared. The highest
protease activity was obtained by fermentation with the Bacillus cultures
but not change by adding skim milk. The cell counting in mixed culture
with ML2 and MKGS showed the growing up together. Slime material
measurement was added to skim milk in mixed cultures was higher.
Keywords: soybean, cheonggukjang
I-68
361
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Anti-viral Effect of Weissella cibaria KM13 Against Avian
Influenza Virus (AIV) H9N2
Jong Bok ROH
1
, Seung-Pyo HONG
1
, Yun-Hee BAEK
3
, Young Ki CHOI
3
,
Haryoung POO
4
, Chul-Joong KIM
4,1
and Moon-Hee SUNG*
1,2
1
BioLeaders Corporation, Daejeon, Korea.
2
Dept. of Bio &Nanochemistry,
Kookmin University, Seoul, Korea.
3
College of Medicine and Medical Research
Institute, Chungbuk National University.
4
Korean Research Institute of Bioscience
and Biotechnology, Daejeon, Republic Korea.
*Corresponding author: jongbok@bioleaders.co.kr
Avian Influenza is flu infection in birds. The virus that causes this infection
in birds can mutate easily and infect humans, This mutation will can start
worldwide epidemic. The study of anti-Avian influenza virus effect
b
ecame
issues because of serious damage of the diseases. Kimchi is a traditional Korean
food, made of vegetable fermented LAB. The study was to characterize
Weissella cibaria KM13 to confirm if the bacterium have anti-viral effects
to AIV. Cell-free supernatants from 21 strains from Kimchi LAB grown in
MRS broth were tested to estimate their anti-viral effects against AIV. Among
21 tested strains, Weissella cibaria KM13 displayed the high anti-viral effects
AIV through Hemagglutination assay (HA). Cell-free supernatant Weissella
cibaria KM13 strain showed 100-fold higher anti-AIV effects than MRS
b
roth
media against AIV H9N2 strain and identified Weissella cibaria KM13
b
y
16sRNA gene sequencing analysis. It was expected that This strain to
p
resent
a anti-viral effects for many subtypes of Avian influenza virus. Futhermore,
Weissella cibaria KM13 may be potential candidates for anti-AIV
development.[This work was supported byTop Brand Project grant from Korea
Research Council of Fundamental Science &Technology and KRIBB Initiative
program (KGM3110912)]
Keywords: avian influenza virus, latic acid bacteria
I-69
Fermentation of Slander Glasswort by Halophile Lactic Acid
Bacteria and Halomonas Subglasiescola
Doo Hyun PARK*, Bo Young JEON, Hana SEO, Seuwon KANG and
Woo Jin LEE
Dept. of Biological Engineering, Seokyeong Univ. Seoul 136-704, Korea.
*Corresponding author: baakdoo@skuniv.ac.kr
Salicornia herbacea L. growing around seashore and saltern is a
hslophyte containing about 0.3% NaCl. We gathered the slander glasswort
from seashore located in Chonranam-do in May 20
th
. The slander glasswort
was a proper plant as the material source for fermentation like Chinese
cabbage. However, the grown slander glasswort is too strong and durable
to eat without process. Protein, fat, sugars and mineral contents are not
significantly different from other edible vegetable. Juice extracted from
the slander glasswort is containing 216.9 mM antioxidant, 652.5 mg/L
of polyphenol and 366.704 mg/L of phenolic acid but is not flavonols.
These may be changed by fermentation of Halomonas subglasiescols and
Shewanella cibaria. The fermentation of extraction and whole plants may
induce the nutritional factors and functional compounds to be improved.
Keywords: slander glasswort, fermentation, halophyte
I-70
Poly(G-Glutamic acid)-Chitosan Nanoparticles Induces
Antigen Specific Humoral and Cellular Immunity
Kyong Soon LEE and Haryoung POO*
korea research institure of bioscience and biotechnology.
*Corresponding author: ghoshow@kribb.re.kr
Nanoparticles are considered to be efficient tools for inducing potent
immune responses by an Ag carrier. In this study, we examined the effect
of Ag-carring biodegradable poly(g-glutamic acid)(g-PGA) chitosan
nanopartles on the induction of immune responses in mice. g-PGA chitosan
nanoparticles strongly induced up-regulation of costimulatory molecules,
and the enhancement of T cell stimulatory capacity in DCs. The
immunization of mice with OVA-carrying nanoparticles induced
Ag-specific CTL activity and Ag-specific production of IFN-g in
splenocytes as well as potent production of Ag-specific IgG Abs in serum.
These results suggest that Ag-carrying g-PGA chitosan nanoparticles are
capable of inducing strong cellular and humoral immune responses and
might be potentially useful as effective vaccine adjuvants for the therapy
of infectious diseases.
I-71
Isolation of Highly Efficient β -Glucosidase Producting Lactic
Acid Bacteria from Kimchi
Jong Gil YOO, Mi-Hee JANG and Myoung-Dong KIM*
School of Bioscience and Biotechnology, Kangwon National University,
Chuncheon 200-701, Korea.
*Corresponding author: mdkim@kangwon.ac.kr
Lactic acid bacteria isolated from a variety of fermented
vegetable(Kimchi) were examined for their capabilities of β
-glucosidase(E.C. 3.2.1.21) production. Based on enzyme activity, most
lactic acid bacteria grown in cellobiose as carbon source showed
significantly higher intracellular β -glucosidase level compared with
extracellular enzyme level. Among the tested 136 strains, genus of
Lactobacillus and Leuconostoc were superior to other group of lactic acid
bacteria for β -glucosidase production. Maximum intracellular β
-glucosidase activity of 5.0±0.2(U/mg) was obtained from
L
eu.
mesenteroides strain isolated from fermented dandelion.
Keywords: biotransformation, B-glucosidase, lactic acid bacteria
I-72
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r

In-vivo monitoring of Cyanide Produced by
P
seudomomonas
aeruginosa Persistently Residing in the Intestine of
Caenorhabditis elegans
Seongwon NAM
1
, Jeesun LIM
1
, You-Hee CHO
2
, Juyoung YOON
1
and
Sungsu PARK*
1
1
Department of Chemistry and Nano Sciences, Ewha Womans University, Seoul
120-750, Korea.
2
Department of Life Science, Sogang University, Seoul 121-742,
Korea.
*Corresponding author: snam@ewha.ac.kr
Pseudomonas aeruginosa (PA) is an opportunistic pathogen that causes
persistent respiratory infections in individuals with cystic fibrosis (CF)
[1]. Many of PA strains produce cyanide (CN
-
) inhibiting cellular
respiration, which presumably aggravates the health condition in CF
patients. Here we report a fluorescence chemosensor enabling imaging
of cyanide produced by PA that persistently resides in the intestine of
the nematode Caenorhabditis elegans. The sensor can detect less than10
mM of cyanide in aqueous solution and showed fluorescent enhancement
only with CN-[2]. Compared to PAO1 strain, PA14 strain produced 4.4
fold more cyanide and killed nematodes faster, while rpoS mutant
produced the highest concentration and was less virulent. These results
suggest that in addition to cyanide production in PA there are many other
virulence factors responsible for the killing of C. elegans. The
chemosensor-based imaging method is highly useful for studying the
infection mechanism of PA as well as rapid screening of antibacterial
compounds against PA. [1] Doring, G. et al. Eur. Respir. J. 16, 749-767
(2000). [2] S. K. Kwon et al. Tetrahedron Lett. 49, 4102 (2008).
I-73
Preparation and Characteristics of Soy Yogurt Fermented
by Bacteria Isolated from Kimchi
Ro-Ui KIM, Jong-Hang SEONG, Young-Guen LEE, Han-Soo KIM and
Dong-Seob KIM*
Department of Food Science &Technology, Pusan National University 50
Cheonghak-ri, Samnangjin-eup, Miryang, Gyeongnam, 602-706, Korea.
*Corresponding author: kds@pusan.ac.kr
Soy curd forming bacteria were isolated from kimchi, a Korean
traditional fermented vegetable food. Among those isolated bacteria, two
strains, 3-4-1 and 4-39-5, made firm curd such as Sundubu. The purpose
of this study was to find the optimum culture conditions of the
b
acteria
for the preparation of firm soy curd. Isolated bacteria were cultured on
MRS broth to find optimum growth condition and the absorbance of
culture broth were measured in 600

. The optimum growth temperature
of 3-4-1 and 4-39-5 were 35

and 30

, respectively. The optimum
growth time of 3-4-1 was 8hr and that of 4-39-5 was 10hr. The hardness
of the soy curd was measured after the bacteria were cultured at the
optimum growth temperature and time. Optimum condition for soy curd
formation was 25

, 24hr for 3-4-1 and 30

, 24hr for 4-39-5. The PH
of soy curd was decreased according to culture time, but the pH showed
fixed value after 18hr. The acidity of soy yogurt was increased according
to culture time, but the acidity also showed fixed value after 36hr. The
bacteria formed firm soy curd with whole soy milk, which has higher
nutritional value than soy milk.
Keywords: soy yogurt, kimchi
I-74
Growth-inhibitory Effects of the Solar Salt-Doenjang on
Cancer Cells
Hae Choon CHANG* and Sun Mi LEE
Dept. of Food and Nutrition, Chosun University, Gwangju, 501-759, Korea.
*Corresponding author: hcchang@chosun.ac.kr
DJI Doenjang was prepared by using Bacillus subtilis DJI starter and
the refined salt or solar salt with 12%(w/v) concentration. The prepared
Doenjangs were fermented and aged for 2 month and 16 month
respectively. MTT assay was used to measure the growth-inhibitory effect
of DJI Doenjang extracts(water, methanol) on BJ(human foreskin normal
cell), HT-29(human colon cancer cell) and AGS(human gastric
adenocarcinoma cell). The anticancer effect increased in both refined
salt-Doenjang and solar salt-Doenjang as fermentation period increased.
In particular, in the case of water extracts, solar salt-Doenjang showed
much better anticancer effect than refined salt-Doenjang. In addition,
apoptosis were observed when the water extracts of the Doenjangs were
treated into AGS. In particular, it was observed that more apoptosis
occured in solar salt-Doenjang treats than refined salt-Doenjang treats.
Keywords: solar salt-doenjang, Bacillus subtilis DJI
I-75
Lactobacillus casei CJNU 0588 Producing Growth Stimulator
for Bifidobacteria
Gi-Seong MOON* and Ji-Eun EOM
Division of Food and Biotechnology, Chungju National University,
J
eungpyeong,
Chungbuk 368-701, Korea.
*Corresponding author: gsmoon@cjnu.ac.kr
Three hundreds of bacterial isolates from natural cheese were screened
for production of bifidogenic growth stimulator by whey fermentation.
From the results, two isolates were selected for the purpose. The culture
supernatants from whey fermentation, effectively enhanced the growth
of Bifidobacterium longum FI10564 by 1.6-1.7 fold compared with a
negative control, non-fermented whey medium. The two isolates were
identified Lactobacillus casei (99% identity) by 16S rRNA gene
sequencing and named Lactobacillus casei CJNU 0421 and CJNU 0588,
respectively. The culture supernatant of Lb. casei CJNU 0588 from whey
fermentation was added to appropriate media where Escherichia coli DH5
α, Enterococcus faecalis KFRI 675 as intestinal bacteria and
Staphylococcus aureus ATCC 14458, Listeria monocytogenes ATCC
19111 as pathogenic bacteria were separately inoculated, to confirm
whether the supernatant selectively stimulate the growth of
b
ifidobacteria.
From the results, none of strain grew fast compared with controls without
the supernatant, indicating metabolite from Lb. casei CJNU 0588
selectively stimulates the growth of bifidobacteria by whey fermentation.
This work was supported by a grant (20080401034067) from BioGreen
21 Program, Rural Development Administration, Republic of Korea.
Keywords: Lactobacillus casei, bifidogenic growth stimulator, whey
fermentation
I-76
363
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Analysis of ces Gene Encoding Cereulide of Temperate Phages
Isolated from Emetic B. cereus
Young-Duck LEE
2
, Tae-Hwa RYU
1
, Hyo-Ihl CHANG
2
and
Jong-Hyun PARK*
1
1
Kyungwon University.
2
Korea University.
*Corresponding author: gene75@empal.com
Bacillus cereus is recognized world widely as an important food-borne
pathogen because of highly environmental contamination rate and is
known to cause clinical infections, food poisoning of toxin-induced
diarrhea and vomiting syndrome. Especially, the emetic toxin called
cereulide is a small, heat and acid stable cyclic dodecadepsipeptide. In
this study, two temperate phages were isolated from emetic B. cereus
and performed in DNA restriction patterns, structural proteins and
morphology analysis. ces gene encoding cereulide was also analyzed
b
y
PCR and sequenced. In results of PCR and sequence, ces gene from
temperate phages by PCR was detected and cereulide synthase B
b
y
BLAST analysis was identified. Sequence alignments of ces gene of
temperate phages and emetic B. cereus showed 99% sequence homology.
Therefore, emetic toxin transfer of B.cereus in nature might be related
with bacteriophage. Other investigators reported emetic toxin gene was
plasmid-borne gene, however, our results did detection of ces gene from
temperate phages. Additional experiments about origin of emetic toxin
needed to be done.
Keywords: temperate phages, emetic B. cereus, cereulide
I-77
Prevalence of tet Genes in Foodborne Tetracycline Resistant
Staphylococcus aureus Isolated from 2003 to 2006 in Korea
Chae Hong RHEE
1
, Il Kwon BAE
1
, Hyon Ji KOO
1
, Koo Youn KIM
1
,
Hyo-Sun KWAK
2
, In-Gyun HWANG
2
, Yong Ho PARK
3
, Kyungwon LEE
4
and Gun-Jo WOO*
1
1
Division of Food Bioscience &Technology, College of Life Sciences
&Biotechnology, Korea University, Seoul, Korea.
2
Division of
M
icrobiology,
Korea Food and Drug Administration, Seoul, Korea.
3
Department o
f

Microbiology, College of Veterinary Medicine, Seoul National University, Seoul,
Korea.
4
Department of Laboratory Medicine and Research Institute of Bacterial
Resistance, Yonsei University College of Medicine, Seoul, Korea.
*Corresponding author: chaehongak@korea.ac.kr (visionkorea@korea.ac.kr)
Introduction: Tetracycline is one of the most commonly used antibiotics
worldwidely, and tetracycline resistance has been the single most common
resistance marker in foodborne microorganisms. In this study, foodborne S.
aureus isolates resisitant to tetracycline were screened to determine the
p
resence
of the resistant tet genes. Materials and Methods: During the period of 2003
to 2006, S. aureus isolates were collected from raw meat (poultry, pork and
beef) and fish samples in Korea. Isolates were tested by the disk diffusion
test and minimal inhibitory concentration (MIC) following the procedures as
described by Clinical and Laboratory Standards Institute. Whole DNA was
extracted by boiling method. The resistance determinant tetM and tetK genes
were screened by PCR assay. Results: Sixty-nine S. aureus isolates (37.1%)
out of 186 showed tetracycline resistance. Among them, tetK and tetM gene
were identified from 3 isolates and 1 isolate, respectively. The isolates having
tet genes had MIC range 16-64 μ g/mL. Conclusions: Tetracycline resistant
S. aureus was prevalent in food marketed in Korea. The results obtained in
this study show that periodical and systematic surveillance is necessary for
inspecting and preventing the spread of foodborne tetracycline resistant isolates.
Keywords: tetracycline resistant Staphylococcus aureus, foodborne
microorganism, tet gene
I-78
Enzymatic Properties of Purified α -Galactosidase from
Mortierella vinacea
Gwi Gun PARK*
2
and Eun Jeong SEO
2
1
Dept. of Food &Bioengineering, Kyungwon University.
2
Dept. of Food
&Bioengineering, Kyungwon University.
*Corresponding author: ggpark@kyungwon.ac.kr
Mortierella vinacea was cultured in 100 mL of medium composed o
f

1.5% Lactose, 0.5% glucose, 0.6% (NH4)2SO4, 1.0% Corn steep liquor,
0.4% KH2PO4, 0.2% MgSO4

7H2O, 0.2% NaCl, 0.12% (NH2)2CO,
0.3% CaCO3 at 30

. α -galactosidase activity from Mortierella sp. was
purified by ion exchange chromatography on CM-sephadex C-50 and gel
filtration chromatography on Sephadex G-100. The purified α
-galactosidase showed a single protein band on SDS. The molecular mass
of the purified α -galactosidase was estimated to be 56 kDa by SDS.
The enzyme was purified approximately 14.57-fold with a yield of 72.54%
compared with the culture filtraion. The purified galactosidase was showed
maximal activity at pH 4.0 and 55

, and was stable in the pH and
temperature ranges of 3.0 to 5.5 and 20

to 60

, respectively. The
enzyme activity was inhibited by Ag2+, Zn2+ and. Hg2+. The enzyme
activity was not affected considerably by treatment with other metal
compounds.
Keywords: purification, α -galactosidase
I-79
Analysis of Bacterial and Fungal Communities in Japanese
and Chinese Fermented Soybean Paste using DGGE
Tae Woon KIM, Jun Hwa LEE, Min Hee PARK and Hae Yeong KIM*
Graduate School of Biotechnology, Kyung Hee Univ. Yongin 446-701,
K
orea.
*Corresponding author: korkimchiman@khu.ac.kr
The microbial diversity in Japanese and Chinese fermented soybean
paste was investigated using a nested PCR-denaturing gradient gel
electrophoresis (DGGE) approach. Five samples of Japanese fermented
soybean paste and three samples of Chinese fermented soybean paste were
used for screening of bacteria and fungi. The DNA extracted was used
as a template for PCR to amplify 16S rRNA and 18S rRNA genes. In
the first step, the nearly complete 16S rRNA and 18S rRNA genes were
amplified using universal primers. Subsequently, these products were used
as a template in a nested PCR to obtain fragments suitable for
DGGE.Tetragenococcus sp. and Staphylococcus sp. strains were found
to dominate the bacteria microbiota in Japanese samples, whereas Bacillus
sp. was detected as predominant species in Chinese samples. In analysis
of fungi, Aspergillus sp. and Zygosaccharomyces sp. were detected in
most of the Chinese and Japanese samples.
Keywords: fermented soybean paste, DGGE
I-80
364
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r

Application of T-RFLP to Monitor the Survival of Probiotic
Lactobacillus plantarum in Mouse Gut
Jong-Hoon LEE*, Myeongjae LEE and Sangmin SHIM
Dept. of Food Science and Biotechnology, Kyonggi University, Suwon 443-760,
Korea.
*Corresponding author: jhl@kyonggi.ac.kr
Chlorogenic-acid-hydrolyzing 22 bacteria were isolated from 7 types
of kimchi, Korean fermented vegetable. They were genotypically identified
as L. plantarum, L. pentosus, and L. brevis. To examine their potential
as probiotics, acid and bile tolerances, antibacterial activity against
foodborne pathogens, and hemolysis type were assayed. L. plantarum
strain KK3 was selected as the prospective probiotic candidate and its
survival in animal gut was monitored by T-RFLP (terminal restriction
fragment length polymorphism) with the freeze-dried cell fed mice for
7 days. The existence of KK3 in the mouse feces was monitored until
31 days and semi-quantitative change of KK3 in the feces was dramatically
detected at T-RFLP profiles.
Keywords: T-RFLP, kimchi, probiotics
I-81
Development of Direct Screening Method for Dextran-forming
Activity of Dextransucrases and its Application to Mutant
Screening
So-Ra LEE, Ah-Rum YI, Myoung-Uoon JANG, Seung-Ho CHOI,
Hong-Gyun LEE, Jung-Mi PARK and Tae-Jip KIM*
Department of Food Science and Technology, Chungbuk National University,
Cheongju, 361-763, Korea.
*Corresponding author: tjkim@cbnu.ac.kr
Dextransucrase (DSase) is a glucosyltranferases that catalyzes synthesis
of a high molecular weight polymer, dextran, from sucrose. DSase can
be screened on phenylethyl alcohol agar medium containing sucrose,
b
ut
this case may be limited in the production of extracellular DSaes. The
huge molecular mass of common DSases causes extremely low level of
heterologous expression systems, which can be one of barriers for its
industrial application. N-terminal truncation of LcDS (
L
euconostoc
citreum DSase) has been tried to generate the minimized mutant enzymes
with considerable activity. For the simple detection of the truncated LcDS,
high-throughput screening techniques should be developed first. To screen
the LcDS mutant with dextran-forming activity, E. coli transformants were
tooth-picked onto LB agar medium containing sucrose. After incubation
for 6~8 hours, soft the agar solution with D-cycloserine was overlaid
on the plate. During incubation at 30

for 12 hours, positive clones were
successfully selected from their capability dextran-forming around the
colony on a plate. In this study, the high-throughput screening method
via direct detection of polymer formation on agar plate developed here
has been successfully applied to the screening of various active truncated
mutants.
Keywords: dextran, dextransucrase, screening method
I-82
A Sensitive Method to Detect Escherichia coli Based on
Immunomagnetic Separation and Aptamers amplification of
Real-Time PCR
Hye-Jin LEE and Min-Kyu OH*
Department of Chemical and Biological Engineering, Korea University, Seoul
136-701, Korea.
*Corresponding author: hyejin703@korea.ac.kr
Aptamers, single stranded nucleic acids, provide a unique opportunity
not only as recognition molecules like antibodies, but also as amplifiable
molecules using polymerase chain reaction (PCR). We report a highly
sensitive detection of E. coli by taking advantage of the aptamer
amplification as well as the specific binding of aptamers. This unique
approach consists of three steps. First, the target E. coli was captured
by antibody-conjugated magnetic beads, the RNA aptamers were
b
ound
onto the surface of captured E. coli in a sandwich way, and the
heat-released aptamers were amplified by using real time
reverse-transcriptase PCR (RT-PCR). This approach has enabled the
detection of ten E. coli in 1 ml sample, thanks to the aptamer amplification
together with a large number of aptamers that bind to the surface of each
E. coli. When compared to the amplification of nucleic acids extracted
from the target microorganisms, this approach can prevent the loss of
target nucleic acids during the sample preparation by obviating the
necessity of cell lysis. This approach also showed a wide dynamic range
from 10
1
to 10
7
E. coli per ml, which can be explained by the exponential
amplification of aptamers.
Keywords: Aptamers, microorganism detection, real-time RT-PCR,
immunomagnetic separation.
I-83
Genome-wide Analysis of Various Xylan-Degrading Enzymes;
Primary, Secondary, and Tertiary Structures
Hong-Gyun LEE, Myoung-Uoon JANG, Jung-Mi PARK, Seung-Ho CHOI,
So-Ra LEE and Tae-Jip KIM*
Department of Food Science and Technology, Chungbuk National University,
Cheongju, 361-763, Korea.
*Corresponding author: tjkim@cbnu.ac.kr
Xylans are major hemicellulose components of plant cell wall where
they are closely associated with the main structural polysaccharide of
cellulose, and with the aromatic polymer such as lignin. The main
xylanolytic enzymes are endo-β -1, 4-xylanase (EC 3.2.1.8), which
hydrolyses the insoluble xylan backbone into shorter soluble
xylooligosaccharides, and β -xylosidase (EC 3.2.1.37), which hydrolyses
the soluble xylooligosaccharides from the non-reducing end to liberate
xyloses. Recently, a number of microbial genome data have been available,
which can provide us valuable knowledge for high-throughput approaches
in biochemistry and molecular biology. Important sequence motifs of
various cellulolytic enzymes, including xylanases and xylosidases, were
comparatively analyzed and found from the microbial genome databases.
Therefore, the relationships between structure and function can
b
e
investigated by structure-based alignment of conserved or variant motif
sequences. In this study, a number of xylan-degrading enzymes genes
were fully compared and several novel meaningful sequence or
structure-motifs were newly mined.
Keywords: xylan-degrading enzymes, xylanases, β -xylosidases
I-84
365
The Korean Society for Microbiology and Biotechnology
2009 International Symposium &
A
nnual Meeting
Modulation of Acarbose Hydrolysis in Various
Cyclomaltodextrinases by Changing Valine Residue within
Extra Sugar Binding Space
Myoung-Uoon JANG, Seung-Ho CHOI, So-Ra LEE, Hong-Gyun LEE,
Jung-Mi PARK and Tae-Jip KIM*
Department of Food Science and Technology, Chungbuk National University,
Cheongju, 361-763, Korea.
*Corresponding author: tjkim@cbnu.ac.kr
Cyclomaltodextrinase (CDase; EC 3.2.1.54) and maltogenic amylase
(MAase; EC 3.2.1.133) belong to Glycoside Hydrolase (GH) family 13,
which is capable of hydrolyzing various substrates such as cyclodextrins,
starch, pullulan and maltooligosaccharides. CDase and MAase can
hydrolyze acarbose, a potent α -glucosidase inhibitor, to glucose and
acarviosine-glucose (PTS; pseudotrisaccharides) and transfer PTS to
various sugar acceptors. It was previously reported that ThMA hydrolyzes
acarbose to glucose and PTS, but ThMA V329S mutant produced glucose,
maltose, and acarviosine instead of PTS. In order to generalize the roles
of Valine residue found in common CDase family enzymes, the same
residue of Bacillus halodurans C-125 or Lactobacillus plantarum WCFS
1 was replaced with Serine residue by site-directerd mutagenesis.
Characterization of BHCDV327S and LPCDV335S verified that the
corresponding valine residue play important role in acarbose hydrolysis
pattern of each CDases. In this work, it has been suggested that substrate
specificities, hydrolyzing activity and transferring activity of CDases can
be modulated by changing amino acid residues, including valine, near
extra sugar binding space (ESBS).
Keywords: cyclomaltodextrinases, site-directerd mutagenesis, acarbose
hydrolysis
I-85
Comparison of Bacterial Community in Fermented Soybean
Paste Made with Traditional and Koji Method using
Denaturing Gradient Gel Electrophoresis
Jun Hwa LEE
1
, Tae-Woon KIM
1
, Min-Hee PARK
1
, Hae-Choon CHANG
2
and Hae-Yeong KIM*
1
1
Graduate School of Biotechnology, School of Biotechnology, Kyung
H
ee
University, Yongin 446-701, Korea.
2
Department of Food and Nutrition, Chosun
University, Gwangju 501-759, Korea.
*Corresponding author: s31041@nate.com
Doenjang is a traditional Korean fermented soybean paste.
D
oenjang
made with koji method is used for industrial product, because this method
is easy to control than traditional method. Eight samples of doenjang
pastes were used for screening of bacteria. DNA extraction from each
soybean pastes was performed using the DNA extraction kit. The extracted
DNA was then used as a template for PCR to amplify 16S rRNA genes.
In the first step, the nearly complete 16S rRNA genes were amplified
using universal primers. Subsequently, these products were used as a
template in a nested PCR to obtain fragments suitable for DGGE.
Tetragenococcus sp., Staphylococcus sp., Enterococcus sp., and Bacillus
sp. were detected in doenjang made with koji method, whereas
Leuconostoc sp., Lactobacillus sp., Tetragenococcus sp., and Bacillus sp.
were detected in doenjang made with traditional method.
Keywords: DGGE, doenjang, koji
I-86
Evaluation of Anti-colitic Effect of Lactic Acid Bacteria in
Mice by cDNA Microarray Analysis
Hoyong LEE*, Young-Tae AHN, Jung-Hee LEE and Chul-Sung HUH
R &D Center, Korea Yakult Co., Ltd., 418-12, Korea.
*Corresponding author: hoyongl@hanmail.net
To evaluate the anti-colitic effect of lactic acid bacteria by cDNA
microarray analysis, a lactic acid bacteria mixture (LM) consisting of
Lactobacillus brevis HY7401, Lactobacillus sp. HY7801 and
Bifidobacterium longum HY8004 was orally administered to dextran
sulfate (DSS)-induced colitic mice and the expression profile of numerous
genes was assessed. DSS treatment caused colitic outcomes such as
inflammation and colon shortening. DSS also up-regulated the expression
of inflammation-related genes: pro-inflammatory and chemotactic
cytokines, including IL-1β, TNF-α, IL-6, CCL2, CCL4, CCL7, CCL24,
CXCL1, CXCL2, CXCL5, CXCL9 and CXCL10, and their receptors
CCR3 and CCR7, and other colitis-related genes such as COX-2, PAP,
MMP family, S100a8, S100a9 and DEFA1. LM treatment inhibited the
mRNA expression of inflammation-related and tissue remodeling genes
induced by DSS as well as the colitic symptoms. LM inhibition of the
DSS-induced expression of the representative inflammatory markers, IL-1
β, TNF-α and COX-2, was supported by quantitative real-time
polymerase chain reaction analysis. These findings suggest that LM
ameliorates DSS-induced colitis by regulating inflammatory-related
cytokines as well as tissue remodeling genes.
Keywords: colitis, microarray, dextran sulfate sodium, lactic acid
b
acteria,
cytokine.
I-87
J
_ Environmetal Microbiology and Engineering
Isolation of Environmental Bacteria Containing an Alkane
Hydroxylase Gene with New PCR Primers for the Detection
of Bacterial alkB Gene
Min-A KWON, Joon Young OH, Hyun Suk KIM, Eun Young YU,
Miae LEE, Bong Keun SONG, Hyuk LEE and Jae Kwang SONG*
Chemical Biotechnology Research Center, Korea Research Institute of Chemical
Technology, Daejeon, Republic of Korea.
*Corresponding author: ajee@krict.re.kr
Alkanes are chemically quite inert, but they are first catalyzed
b
y
microbial oxygenases in the bacterial aerobic degradation of alkanes. Such
alkane hydroxylases are promising as versatile biocatalysts, which carry
out a wide range of useful oxidation reactions for the production of fine
hemicals and pharmaceuticals. Highly homologous regions of alkane
hydroxylase gene (alkB) were used to design PCR primer sets. The 25
alkB sequences were classified into three groups and four regions were
selected. A number of bacterial strains, which had been previously isolated
from Taean were examined using four different PCR to screen the positive
alkB gene . The PCR products of the expected length were obtained from
58 strains and sequenced. All the sequences revealed the DNA sequence
identity ranged from 67% to 99%. The 16S rRNA sequences of
alkB-positive strains were determined. We are now attempting to obtain
the entire sequence of the alkB genes using the the results of PCR screening
for the alkB gene.
Keywords: PCR screening, alkane-degrading bacteria, alkane hydroxylase
J-1