Biomimetic Synthesis of Metal Oxide Nanoparticles by using
Peptides and Proteins
John Mwale and Woo-Seok Choe *
Department of Nano Science and Technology, SKKU Advanced Institute of Nano-Technology (SAINT), Sungkyunkwan
University, Suwon, Gyeonggi-do, Korea, *
Abstract
A biomimetic approach is employed for the in vitro synthesis of 2. Theory
Titanium (IV) Oxide nanoparticles, TiO2, at ambient processing
conditions by using a disulphide bond-constrained affinity peptide
STB1 (-CHHKKPSKSC-), and a recombinant protein, LacI-STB1
cognizant of TiO2. Both STB1 peptide and LacI-STB1 protein were
able to catalyze the room temperature hydrolysis and condensation
of a water–soluble TiO2 precursor, Titanium Bis (AmmoniumLacta
to) Dihydroxide, TiBALDH, to form TiO2 nanoparticles which
were subsequently quantified and characterized by Scanning
Electron Microscope (SEM), and Energy Dispersive Spectrometer
(EDS).
1. Introduction
Ever since the pioneering research of Morse and co-workers
illustrating the use of Silicatein (a protein derived from marine
sponge) for the in vitro synthesis of Titania (TiO2) [1], there has
been an increasing interest in methods that mimic
bio-mineralization not only due to their advantage in producing
inorganic materials at ambient processing conditions, but also
because of the ability to control the morphology and polymorph of
the resulting nanocrystals in a predictable manner through the use
of biomolecules that have been specifically selected to recognize a
chosen surface through a biopanning process. In this respect, the
peptide/protein-directed formation of TiO2 nanoparticles is an
attractive alternative to conventional synthetic routes such as
Sol-Gel method [2], Micelle and Inverse Micelle [3], Chemical
Vapor Deposition [4], Hydrothermal [5], etc. were shape control is
achieved by employing various surfactants identified through a
trial-and-error process [6]. TiO2 has a wide array of applications
ranging from pigments [7], sunscreens [8], and paints [9], to
photovoltaics, photocatalysis, photo-electrochromics, and sensors
[10].
Previously in our group we identified STB1, a peptide cognitive
of TiO2 through a phage surface display technique [11], and then
fused this peptide to the C-terminus of a DNA-binding protein Lac
repressor (LacI) to form a designer protein LacI-STB1 which was
then subjected to binding tests. Results showed that it has a high
affinity for TiO2 nanoparticles [12]. In the present study; we used
this engineered designer protein to induce the formation of TiO 2
nanoparticles.
The exact mechanisms by which biomolecules control the
inorganic material formation are still largely unknown [13].
However, Cole and Valentine [14], proposed a mechanism in
which they suggested that the self-assembled structures of
biomolecules can act as a template on which two metal atoms can
be complexed. This complexation then brings these atoms in close
proximity thereby inducing condensation. Another mechanism was
proposed by Jiang et al. [15] to explain the formation of TiO2 by
protamine protein. According to them, the positively charged
protein molecules can adsorb and concentrate the negatively
charged titanium precursor via electrostatic and hydrogen bonding
interactions. With an increase of precursor concentration around
the protein molecules the titanium atoms are brought into close
proximity. Hydrogen bonding between Ti–OH of the titanium
precursor and =NH of the protein induces nucleophilic substitution
of a Ti–O oxygen atom on another adjacent titanium atom, and the
polycon-densation reaction occurs subsequently. Complexation of
titanium with protein molecules offers some additional contribution
to the acceleration of the polycondensation.
As the condensation proceeds, each protein molecule can be
regarded as a globular nucleating center to induce Titania
formation. The Titania particles build up around the protein
molecules, and then aggregate with other adjacent Titania particles
through the condensation reaction between the surface hydroxyl
groups, ultimately resulting in the formation of Titania
nanoparticles.
3. Experiments
3.1 Purification of LacI-STB1 Protein
After fermentation of E. coli BMH8117 (containing plasmid
pWB1000) in dYT media, cells are harvested by centrifugation and
washed with CSB buffer at pH 7.2. A mechanical extraction
method is used for cell lysis, followed by removal of cell debris
through centrifugation. The supernatant is subsequently subjected
to 20% and 35% (NH)2SO4 fractionation to precipitate out
contaminating proteins. Dialysis and Cation exchange
chromatography are the next steps in obtaining pure LacI-STB1
protein. The purified protein is then characterized using SDS-Page
and Western Blotting methods to determine purity.
3.2 TiO2 Precipitation Assay for STB1 and LacI-STB1
For the biomineralization experiments, STB1 peptide
 or LacI-STB1 protein in PBS buffer is mixed with the hydrolysis and condensation of a TiO2 precursor, and assist in
TiO2 precursor (TiBALDH), and left to react for 90 the nucleation and growth of TiO2 nanocrystals. This research
minute at room temperature. provides an interesting insight into the bio-enabled synthesis of
3.3 TiO2 Quantitation Assay inorganic materials for energy applications.
A calorimetric method, the 5-Chlorosalicylic acid assay References
is used to find the amount of TiO2 synthesized. A [1] L.Sumerel, W.Yang, D.Kisailus, J.C. Weaver, J. H. Choi, D. E. Morse,
Chem Mater., 15, 4804, 2003.
complex is formed between the TiO2 nanoparticles and [2] A.C. Pierre and G. M. Pajonk Chem. Re. 102, 4243, 2002.
the acid which can then be detected by using a [3] S. S. Hong, M.S. Lee, S.S. Park, and G.D. Lee, Catal. Today ,
87, 99, 2003.
Spectrophotometer. [4] S. Seifried, M. Winterer , H. Hahn, Chem. Vap. Deposition ,
6, 239,2000
[5] M. Andersson, L. Oesterlund, S. Ljungstroem, A.J. Palmqvist,
4. Results and Discussion Phys. Chem. B 2002, 106, 10674.
Titanium dioxide nanoparticles are synthesized from [6] Y. Li, C. Chiu, and Y. Huang, Pure Appl. Chem.,83, 2011
[7] G. Pfaff and P.Reynders, Chem. Re.,, 99, 1963, 1999.
precursor molecules at benign processing conditions with the [8] A.Salvador, M.C. Pascual-Marti, J.R. Adell, A. Requeni, J.G. March,
use of STB1 peptide and LacI-STB1 protein. Fig.1 shows the J. Pharm. Biomed. Anal., 22, 301,2000
[9] J.H. Braun, A. Baidins, R.E. Marganski, Prog. Org. Coat.,
images of LacI-STB1 induced TiO2 nanoparticles at different 20, 105,1992.
concentrations after 90minutes. The amount of Titania formed [10] A. Millis and S.J Le Hunte, Photochem. Photobiol., A, 108,1, 1997.
[11] C. Haibin, X. Su, K.G. Neoh, W. S. Choe, Anal. Chem., 78,
as shown in Table.1 clearly shows that the TiO2 formation 4872-4879,2006
process is dependent on the concentration of the biomolecules [12] C. Haibin, X. Su, K. G. Neoh, W. S. Choe, (2009), Adv. Mater., 19,
1186-1192
in solution.
[13] S. Mann. Biomineralization: Principles and Concepts in
The SEM images Fig.2 (b), (c), and (d), confirm the Bioinorganic Materials Chemistry, Oxford University Press, Oxford,
bio-enabled formation of the nanoparticles. EDS spectrum 2001
[14] K.E. Cole and A.E. Valentine, Biomacromolecules, 8, 1641 1647, 2007
Fig.2 (b) reveal TiO2 peaks showing that TiO2 was
successfully synthesized. The SEM micrographs also reveal
the morphology of the Titania nanoparticles. When STB1
peptide is used as the biocatalyst, the resultant nanoparticles
are much smaller in size, and all of them exhibit spherical Fig.1 Images (a) Image of TiO2 induced by 3.6mg/mL LacI-STB1
shapes. For the case of the protein, LacI-STB1, the (b) Image of TiO2 induced by 6mg/mL of LacI-STB1
nanoparticle are larger, and shapes are different depending on
the concentration of protein used.
Unlike nanoparticles produced by STB1 peptide, the
nanoparticles synthesized by LacI-STB1 protein are not all
spherical in shape. At a protein concentration of 3.6mg/mL,
the Titania nanoparticles have various shapes; some are 6. (b)
spherical while others are square, and rod-like. When the
protein concentration is increased to 3.6mg/mL,
large,strip-like structures of Titania are formed. The different
morphologies can be attributed to to the speed at which
hydrolysis, condensation of precursor molecules occur at
different biocatalyst concentration. Low concentrations of
(c) (d)
peptide or protein favor faster nucleation and growth of the Fig. 2. SEM images of TiO2 (a) TiO2 synthesized by
nanocrystals, resulting in relatively smaller particles. In 10mg/mL of STB1 peptide (b) EDS image of TiO2
contrast, as the concentration is increased, the nucleation and synthesized by STB1(c) TiO2 synthesized by 3.6mg/mL
of LacI-STB1 Protein (d) TiO2 synthesized by 6mg/mL
growth of crystals occur at a much slower rate, thereby
of LacI-STB1 protein
allowing sufficient time for the crystals to grow, resulting into
Protein/peptide Concentration Amount of TiO2
much bigger particles.
(mg/mL) synthesized
STB1 10 204
5. Summary
LacI-STB1 3.6 156
In this work, it has been shown that a biomimetic method can be
LaI-STB1 6.0 230
used to synthesize TiO2 nanoparticles at room temperature with the
aid of STB1 peptide and LacI-STB1 recombinant protein. The Table 1. Amount of TiO2 induced by peptide and Proteins
biomolecules self-assemble into structures that act as templates for
Morphology control of TiO2 nanoparticles in biomimetic synthesis using affinity peptide

저자 Jae Wook CHO, Ming ZHANG, John MWALE, Woo Seok CHOE
(Authors)

출처 한국생물공학회 학술대회 , 2010.10, 206-206 (1 pages)

(Source)

발행처 한국생물공학회
(Publisher) The Korean Society For Biotechnology And Bioengineering

URL http://www.dbpia.co.kr/Article/NODE02281680

APA Style Jae Wook CHO, Ming ZHANG, John MWALE, Woo Seok CHOE (2010). Morphology control of
TiO2 nanoparticles in biomimetic synthesis using affinity peptide. 한국생물공학회 학술대회,
206-206.

이용정보 14.138.203.***
(Accessed) 2017/06/02 22:07 (KST)

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Copyright of all literary works provided by DBpia belongs to the copyright holder(s)and Nurimedia does not guarantee
contents of the literary work or assume responsibility for the same. In addition, the literary works provided by DBpia may
only be used by the users affiliated to the institutions which executed a subscription agreement with DBpia or the
individual purchasers of the literary work(s)for non-commercial purposes. Therefore, any person who illegally uses the
literary works provided by DBpia by means of reproduction or transmission shall assume civil and criminal responsibility
according to applicable laws and regulations.
KSBB, Abstracts of Current Biotechnology and Bioengineering(XXVII) : October, 2010
PFM 21 Inhibitory Effects of Pine gnarl Water Extracts on
Atopic Disease
Chung-Jo LEE, Eu-Jin SONG, Ji-Hee KWAK, Koth-Bong-Woo-Ri
KIM, Ji-Yeon JUNG, Min-Ji KIM, Moon-Kyung CHOI, Dong-Hyun
1
KIM, Chan SUNWOO, Tae-Wan KIM and Dong-Hyun AHN
Dept. of Food Science and Technology, Pukyong National University,
Busan 608-737. 1Dept. of Food Science and Technology, Andong
National University, Gyeongbuk 760-749.
Many investigators have tried to look for atopic dermatitis medicine from
natural resources lately because the occurrence of atopic dermatitis has
been continuously increasing. Therefore, an aim of this study is to
examine inhibitory effects of pine gnarl water extracts (PGWE) on
atopic dermatitis. To conduct this experiment, PGWE was added to
splenocytes obtained from OVA-immunized mice for an in-vitro test. In
addition, an anti-atopic dermatitis test was carried out by spreading
PGWE on the dorsal skin of 2,4dinitrochlorobenzene (DNCB)-induced
balb/c mice. It was verified that the IFN-γ, IL-4 and total IgE secretion
was suppressed when PGWE was added to splenocytes obtained from
OVA-immunized mice. For anti-atopic dermatitis test, PGWE alleviated
symptoms of the erythema, crust and dryness in DNCB-induced mice.
Furthermore, the IFN-γ, IL-4 and total IgE secretion of the group
treated with PGWE in splenocytes and sera considerably diminished
compared to the positive control. Through these results, we can conclude
that PGWE can inhibit allergy and atopic dermatitis by modulating
IFN-γ, IL-4 cytokine and IgE secretion.
Keywords : Atopic dermatitis , Allergy , Pine gnarl
PFM 23 Protective Effects of Hispidin on Hydrogen Peroxide
Induced Oxidative Stress in MIN6N Pancreatic beta-Cells
Jung Hyun LEE, Young-Rae KIM and Eock-Kee HONG
Department of Bioengineering and Technology, Kangwon National
University, Chuncheon 200-701, Korea.
Reactive oxygen species (ROS) have been shown to cause DNA damage,
denaturation of protein, loss activity on anti-oxidative enzyme and lipid
peroxidation. Thus, ROS are associated with tissue damage and the prime
contributing factors for inflammation, diabetes, aging, cancer, hypertension
and arteriosclerosis. Some mushrooms, including Phellinus linteus, commonly
produce a bundle of yellow antioxidant pigments that are composed of hispidin
derivatives and polyphenols. Hispidin, metabolite of Phellinus linteus, is a
potent antioxidant and inhibitor of protein kinase C (PKC). In this study, it was
investigated whether hispidin would protect MIN6N pancreatic β-cells from
oxidative stress by hydrogen peroxide. Treatment of MIN6N β-cells with
0.5mM hydrogen peroxide for 4 hours could cause a significant cell viability
loss and an increase in apoptotic cells. Pre-treatment of MIN6N β-cells with
hispidin for 24 hours reduced cell viability loss and decrease in apoptotic cells.
In addition, 70μM hispidin significantly scavenged the intracellular ROS and
inhibited apoptosis induced by hydrogen peroxide. In addition, the generation
of thiobarbituric acid reactive substance (TBARS) was inhibited in the
presence of hispidin in a dose-dependent manner. Also, 70μM hispidin
significantly increased insulin secretion. These results suggested that hispidin
might reduce oxidative stress damage of MIN6N β-cells by hydrogen peroxide.
Hispidin may be effective in preventing MIN6N β-cells from the toxic action of
reactive oxygen species in diabetes.
Keywords : Hispidin , pancreatic beta-cells , ROS
- 206 -
IP: 14.138.203.*** | Accessed 2017/06/02 22:07(KST)
PFM 22 Immunopotentiating Activity of Sargassum Thunbergii
Polysaccharide
Eu-Jin SONG, Chung-Jo LEE, Koth-Bong-Woo-Ri KIM, Ji-Yeon JUNG,
Min-Ji KIM, Moon Kyung CHOI, Dong-Hyun KIM, Chan SUNWOO
and Dong-Hyun AHN
Dept. of Food Science and technology, Pukyong National University,
Busan, 608-737.
Seaweeds have been attracted recently as multifunctional foods for
maintaining human health. So, this study was carried out to verify immune
enhancing activities of polysaccharide from Sargassum thunbergii(STP),
edible brown seaweeds. Mice (n=5) were orally administered with STP
every day for 4 weeks. Peyer’s patch, spleen and peritoneal macrophages
were harvested from STP supplemented mice. It was confirmed that the
oral administration of 10 and 100 mg/kg.BW STP enhanced proliferation
abilities of peyer’s patch cells, splenocytes, and peritoneal macrophages.
The IFN-γ, IL-6 as well as IgA secretion of peyer’s patch cells were
significantly increased by STP. In addition, the splenocytes from STP
administered mice produced substantially more IFN-γ and peritoneal
macrophages secreted more IL-6 and IL-1β compared to PBS administered
mice. In conclusion, it is considered that the oral administration of STP
could strengthen immune reactions through stimulating an intestinal
immunity.
Keywords : Sargassum thunbergii , Immunopotentiating activity ,
Peyer's patch , Macrophage , Spleen
PFM 24 M o rpho lo g y co ntro l o f TiO 2 nano particles in
biomimetic synthesis using affinity peptide
Jae Wook CHO, Ming ZHANG, John MWALE and Woo Seok CHOE
Department of Chemical Engineering, Sungkyunkwan University,
Suwon 440-746, Republic of Korea.
Peptide aptamers that recognize inorganic materials are attracting increasing attention in the field of
nanobiotechnology. This is because these peptides can be used not only to allocate various molecules on
patterned substrates but also to form nanocrystals of inorganic materials under ambient conditions.1 TiO2
nanoparticles are widely used for many applications including photo-catalysis, photo-voltaics, energy
storage and so on. The peptide-directed room-temperature formation of TiO2 nanoparticles can be an
2
attractive alternative to higher-temperature synthetic methods. However, the underpinning mechanism for
peptide driven biomimetic TiO2 synthesis remains still unclear and it has a long way to go before producing
crystalline TiO2 with controlled nanostructure conducive to practical device applications. In the present
study, we used STB1 (-CHKKPSKSC-, a constraint heptapeptide cognitive of TiO2) and its derivatives (i.e.
3
point mutants of STB1 and STB1 fused with the tail peptide sequence of R5 reported elsewhre ) to explore
a strategy for controlling the morphology (i.e. size and shape) of TiO2 precipitate in various conditions (e.g.
peptide concentration, buffer composition, pH of buffer solution). Furthermore, the effect of peptide local
structure (i.e. linear vs constraint form) on the morphology of biomineralized TiO2 precipitate was
investigated. All the nanoparticles synthesized were characterized by energy dispersive spectroscopy
(EDS), X-ray diffraction (XRD), scanning electron microscope (SEM) and transmission electron
microscope (TEM). It was found that kinetics of TiO2 biomineralization is a key to determine the size of
TiO2 nanoparticles: faster kinetics give rise to smaller nanoparticles. However, the kinetics of TiO2 particle
formation exhibited little effect on the shape of particles, hence resulting in spherical nanoparticles with
interconnected one another. Interestingly, the local structure of peptide molecules was found to affect the
shape of TiO2 precipitate, giving rise to spherical nanoparticles with the use of constrained peptide
sequences and planar interconnected precipitates with linear form.
Keywords : biomimetic synthesis , constraint peptide , affinity peptide
KSBB, Abstracts of Current Biotechnology and Bioengineering(XXVIII) : April, 2011
PEB 21 Factors affecting the growth of microalgae, Tetraselmis
suecica for the oil accumulation
Sugeun GO, Seok-Joo LEE, Hyejin KIM and Sung-Koo KIM
Dept. of Biotechnology, Pukyong National University, Busan 608-737.
M ircoalgae require proper conditions for the cell grow th. Since
microalgae are photosynthetic, proper light source, carbon dioxide
concentration, water, and inorganic salts are required for optimal
growth conditions. In this study, various light intensities and nitrate
concentrations were tested with conditions of 20℃, 24:0 L:D cycle and
2.5 L/min aeration rate in 20 L batch culture. It took 5 days to reach the
stationary phase for the culture of T. suecica with the light intensities of
108.9 and 133.1 μmol/m2/s with biomass of 0.89 and 0.88 g dcw/L,
respectively. And, biomass productions of 1.07 and 1.00 g dcw/L were
obtained with the nitrate concentrations of 18.55 and 24.74 mg/L,
respectively. Nitrate was consumed by T. suecica of all tested groups
within 5 days after the inoculation. Two-stage culture process through
nitrate depletion led to increase oil contents from 7.6 % to 17.3 % (w/w)
and contents of C16∼18 fatty acids from 540.2 mg/g oil to 720.5 mg/g
oil at 4th day after nitrate depletion. The biomass production and oil
content of T. suecica was enhanced by the optimization of growth
conditions and 2-stage culture process.
Keywords : microalgae , oil , two stage culture , biodiesel
PEB 23 Biomimetic synthesis of TiO 2 -WO 3 hybrid metal
oxide nano particles induced by a designer protein
John MWALE, Noo-ri CHOE and Woo-Seok CHOE
Department of Chemical Engineering, Sungkyunkwan University,
SKKU Advanced Institute of Nanotechnology,Suwon 440-746, Korea.
Ever since the pioneering research of Morse and co-workers illustrating the
use of Silicatein (a protein derived from marine sponge) in the in vitro
synthesis of Titania1, there has been an increasing interest in methods that
mimic bio-mineralization due to their advantage in producing inorganics at
ambient conditions.TiO 2 is commonly used as photo-catalyst due to its
non-toxicity, chemical stability, cheapness, and its high efficiency. However,
owing to its relatively large band gap (~3.2 eV) its use is limited to the UV
light region. By coupling the two metal oxide semiconductors TiO 2 and WO 3,
the reduction of band gap energy can be achieved, which will allow the hybrid
to be used in the visible light region.Previously in our group we identified
STB1 (-CHKKPSKSC-), a constrained hepta-peptide cognitive of TiO 2
through a phage surface display technique,2 and then fused this peptide to the
C-terminus of a DNA binding protein LacI to form a designer protein
LacI-STB1 which was then subjected to binding tests. Results showed that it
3
has a high affinity for TiO 2 nanoparticles. In the present study; we used this
engineered designer protein to induce the formation of TiO 2-WO 3 hybrid
metal oxide nano-particles. The synthesized hybrid nano-particles were
subsequently quantified and characterized by Scanning Electron Microscope
(SEM) and EDS. Compared to using peptides, this research presents a
cheaper method for the bio-mimetic synthesis of hybrid metal oxides.
Keywords : Bio-mimetic synthesis , Designer protein , Hybrid metal oxide
nano particles , Photo catalysis
IP: 14.138.203.*** | Accessed 2017/06/02 00:40(KST)
PEB 22 Biobutanol Production from Non-edible Biomass
using Clostridia
Julia LEE, D ukki K IM , M oon-H o EO M , Sang-H yun LEE and
Jung-Hee CHO
R&D center, GS Caltex Corporation, 104-4 Munji-dong, Yusung-gu,
Daejeon 305-380.
The production of acetone-butanol-ethanol (ABE) by clostridium, which
is a Gram-positive, obligate anaerobic microorganism, is recently
studied for bio-based chemicals and fuels. However, raw material cost
is the most critical for the economics. Medium cost usallaly accounts
for 65 % of total direct expense. In this study, non-edible biomass such
as palm residues was utilized for the production of butanol as a main
carbon source. The final ABE titer of palm biomass hydrolysate
decreased campared to glucose medium. On the other hand, total ABE
yield of palm biomass hydrolysate was enhanced compared to glucose
m edium . This w ork w as supported by the Energy Efficiency &
Resources of the Korea Institute of Energy Technology Evaluation and
Planning (KETEP) grant funded by the Korea government Ministry of
Knowledge Economy.
Keywords : Non-edible biomass , Biobutanol , Fermentation
PEB 24 Ionic Liquid-mediated Extraction of Lipids from
Algal Biomass
Young-Hoo KIM, Yong-Keun CHOI, Jung Su PARK, Seong Min LEE,
Yung-Hun YANG, Hyung Joo KIM and Sang Hyun LEE
Department of Microbial Engineering, Konkuk University, Seoul 143-701.
The lipids of algal biomass were extracted by using mixtures of ionic
liquids (ILs) and methanol, and the fatty acid profiles of extracted lipids
were also characterized in this work. ILs which cannot dissolve lipids
were chosen selectively to extract lipids from algal biomass. Mixtures
of ILs and methanol successfully dissolved biomass leaving lipids
insoluble. Total contents of lipids extracted from commercial and
cultivated Chlorella vulgaris were 10.6 and 11.1% by conventional Bligh
and Dyer’s method, respectively, while the mixture of [Bmim][CF3SO 3]
and methanol could extract 12.5 and 19.0% of lipids. Multi-parameter
regression by linear solvation energy relationship showed that dipolarity/
polarizability and hydrogen bond acidity of ILs play an important role
in the extraction of lipids from biomass. Fatty acid profiles of extracted
lipids showed wide range distribution from C14 to C24. Especially,
C16:0, C16:1, C18:2, and C18:3 fatty acids were dominant in the lipids
extracted by using ILs. It means that the extracted lipids from C.
vulgaris can be used as a source of biodiesel production. These results
show high efficiency of ILs-mediated extraction of lipids from algal
biomass.
Keywords : Chlorella vulgaris , algal biomass , ionic liquid , biodiesel
Patterned biomineralization of TiO2 using DNALacI scaffold

저자 Jae Wook CHO, John MWALE, Byung Hoon KIM, Sung Ha PARK, Woo Seok CHOE
(Authors)

출처 한국생물공학회 학술대회 , 2010.10, 201-201 (1 pages)

(Source)

발행처 한국생물공학회
(Publisher) The Korean Society For Biotechnology And Bioengineering

URL http://www.dbpia.co.kr/Article/NODE02281659

APA Style Jae Wook CHO, John MWALE, Byung Hoon KIM, Sung Ha PARK, Woo Seok CHOE (2010).
Patterned biomineralization of TiO2 using DNALacI scaffold. 한국생물공학회 학술대회, 201-
201.

이용정보 14.138.203.***
(Accessed) 2017/06/02 22:07 (KST)

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가 비영리적으로만 이용할 수 있습니다. 그러므로 이에 위반하여 DBpia에서 제공되는 저작물을 복제, 전송 등의 방법으로 무단
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Copyright Information
Copyright of all literary works provided by DBpia belongs to the copyright holder(s)and Nurimedia does not guarantee
contents of the literary work or assume responsibility for the same. In addition, the literary works provided by DBpia may
only be used by the users affiliated to the institutions which executed a subscription agreement with DBpia or the
individual purchasers of the literary work(s)for non-commercial purposes. Therefore, any person who illegally uses the
literary works provided by DBpia by means of reproduction or transmission shall assume civil and criminal responsibility
according to applicable laws and regulations.
 한국생물공학회, 생물공학의 동향(XXVII)：2010. 10

PFM 01 Encapsulation of Fish Oil using Iota Carrageenan PFM 02 Enzymatic Synthesis and Applications of Hydroquinone
and Sodium Casein Glucoside using Leuconostoc mesenteroides Dextransucrase
Hye-Sook NA and Ki-Young LEE 1 Eun-Seong SEO, Go-Eun KIM 1, Jin KANG and Sun-Hwa JUNG 1
Department of Advanced Chemicals Graduate School, Chonnam D epartm ent of Biological Sciences and Technology, Chonnam
1
National University, Gwangju, 500-757, KOREA. University, Gwangju, 500-757, Korea. Interdisciplinary Program of
1
Faculty of Applied Chemical Engineering, and Institute for Catalysis Graduate School for Bioenergy and Biomaterials, Chonnam National
Chonnam Nat. Univ., KOREA. University, Gwangju 500-757, Korea.

The study about masking odors and reducing oxidation was performed. We synthesized a hydroquinone glucoside (HG) as a potential skin
Encapsulating properties of iota carrageenan (IC) and sodium casein whitening agent using Leuconostoc mesenteroides dextransucrase with
(SC) for the coating of the fish oil were evaluated. Retention of fish oil hydroquinone (HQ) as an acceptor and sucrose as a donor. The product
(oil red o staining) after freeze-drying was observed by measuring was purified using butanol partitioning and silica-gel column
absorbance. Highest amount of fish oil (70 ± 0.3% of initially added chromatography. The structure of the purified HG was determined by
amount) was retained in a matrix consisting of IC and SC, in the ratio of NMR. HG was identified as 4-hydroxyphenyl-α-D-glucopyranoside.
0.1:1:1 (w/w). Particle size varied from 2.0 to 3.2 μm. The fishy odor of The optimum condition of HG synthesis, determined using a response
raw fish oil was considerably reduced by encapsulation of IC and SC. A surface methodology, was 450 mM HQ, 215 mM sucrose, and 0.55
mixture consisting of IC-SC is an efficient encapsulant for fish oil U/mL dextransucrase; the final HG produced was 544 mg/L HG
encapsulation. potently inhibited oxidation. The IC50 of diphenylpicryl-hydrazyl
scavenging activity was 3.85 mM indicating a higher anti-oxidant
Keywords : iota carrageenan , sodium casein , fish oil , encapsulation activity compared to β-arbutin (IC50 = 6.04 mM). HG-mediated
inhibition of lipid peroxidation was 3.51% that of HQ (100%) and much
higher than that of β-arbutin (0.81% of HQ). In addition the IC50 value
of nitrite scavenging activity was 14.76 mM showing a superior
scavenging activity to that of β-arbutin (IC50 = 27.09 mM)

Keywords : Dextransucrase , Hydroquinone glucoside , Tyrosinase

PFM 03 Patterned biomineralization of TiO 2 using DNA- PFM 04 Antioxidative activity of Suaeda japonica extracts
LacI scaffold harvested at different seasons
Jae Wook CHO, John MWALE, Byung Hoon KIM 1 , Sung Ha PA RK 1 Jong-Il CHOI, Jae-Hun KIM, Beom-Suk SONG and Ju-Woon LEE
and Woo Seok CHOE Korea Atomic Energy Research Institute.
School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
1
Department of Physics, Sungkyunkwan University, Suwon 440-746, Republic of Korea. This study was conducted to investigate the physiological activities of
extracts from Suaeda japonica harvested in different season for its
Nanomaterials exhibit a wide spectrum of properties stemming from their nanoscale dimension and morphological features. possibility as a functional material in food or cosmetic composition.
Biomolecules have attracted increasing attention in the field of nanobiotechnology due to their exquisite specific recognition ability The total mineral content of S. japonica harvested in summer was about
1)
toward various inorganic materials including metals and semiconductors. Many biomolecules were shown to mediate the formation 89.8 g/kg, and it comprised a little more content than one in winter (86.7
of inorganic nanocrystal structures from their precursors under ambient condition through a process called biomineralization. g/kg). The Na content of S. japonica did not show a remarkable contrast
However, most of biomineralized inorganic nanostructure reported elsewhere showed deductive morphology (i.e. morphology is on harvest season whereas the K and Ca contents of summer were
determined after trial and error based biomineralization) rather than inductive morphology (i.e. morphology is predicted prior to decreased to half or increased to double in winter. In addition, the
biomineralization). This raises a need for an alternative approach capable of giving rise to a patterned structure precisely mimicking antioxidative activity of each extract from S. japonica changed
a pre-designed biomolecular scaffold which is conducive to designing practical devices with a desired performance in a predictable depending on harvest season. For S. japonica harvested in summer, the
manner. To explore a novel strategy for achieving inductive nanostructure, this study aims to synthesize TiO2 nanoparticles and its ethyl acetate extract showed the highest DPPH radical scavenging
patterned nanostructure capable of exhibiting new physical properties at nanometer scale which would otherwise be difficult to realize activity, but in winter the butanol extract fraction had the highest value.
by the conventional metal oxide nanofabrication. A core strategy for formation of TiO2 nanoparticles and subsequent simple patterned However, FRAP values were the highest in butanol extracts from S.
nanostructure is to mimic exquisite biomineralization mechanism evolved by living entities in nature. For this, we created a designer japonica harvested in summer and winter. Total phenolic contents in the
protein LacI-STB1 harboring TiO2 cognitive peptide at the C-terminus of LacI while leaving DNA binding domain (DBD) of LacI extracts were in proportion to the antioxidative activities.
intact. By using LacI-STB1 as a linker to position TiO2 molecules on the plasmid DNA (pDNA) harbouring lacO at a specific
position, it was demonstrated that TiO2 nanoparticles could be positioned at a specific position on pDNA/LacI-STB1 scaffold (i.e. a Keywords : Suaeda japonica , antioxidant , harvest season
simple inductive structure). Ongoing study will further test a possibility of biomineralizing TiO2 at programmed positions with the use
of DNA/Lac-STB1 scaffold as an inductive structure where LacI-STB1 acts as a mineralizing catalyst as well as an anchor to position
such mineralized TiO2 particles. In parallel, various 1-, 2- and 3-D structures of DNA by harnessing DNA origami technology were
synthesized and LacI-STB1 binding to these diverse DNA scaffolds was confirmed, highlighting a potential of realizing complicated
inductive nanostructured metal oxide with the use of pre-designed biomolecular scaffolds.

Keywords : Biomimetic synthesis , patterned nanoparticle , LacI protein , Lac repressor , DNA origami

- 201 -

IP: 14.138.203.*** | Accessed 2017/06/02 22:07(KST)

Kinetics and Morphology Control of TiO₂ by Biomineralization using Affinity Peptide

저자 Noori CHOI, John MWALE, Woo-Seok CHOE

(Authors)

출처 한국생물공학회 학술대회 , 2011.10, 235-235 (1 pages)

(Source)

발행처 한국생물공학회
(Publisher) The Korean Society For Biotechnology And Bioengineering

URL http://www.dbpia.co.kr/Article/NODE02256744

APA Style Noori CHOI, John MWALE, Woo-Seok CHOE (2011). Kinetics and Morphology Control of
TiO₂ by Biomineralization using Affinity Peptide. 한국생물공학회 학술대회, 235-235.

이용정보 14.138.203.***
(Accessed) 2017/06/02 22:06 (KST)

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Copyright Information
Copyright of all literary works provided by DBpia belongs to the copyright holder(s)and Nurimedia does not guarantee
contents of the literary work or assume responsibility for the same. In addition, the literary works provided by DBpia may
only be used by the users affiliated to the institutions which executed a subscription agreement with DBpia or the
individual purchasers of the literary work(s)for non-commercial purposes. Therefore, any person who illegally uses the
literary works provided by DBpia by means of reproduction or transmission shall assume civil and criminal responsibility
according to applicable laws and regulations.

PFM 09 K inetics and M o rpho lo g y Co ntro l o f TiO 2 by
Biomineralization using Affinity Peptide
Noori CHOI, John MWALE and Woo-Seok CHOE 1
SKKU Advanced Institute of Nanotechnology(SAINT), Sungkyunkwan
University, Suwon 440-746, Republic of Korea.
1
School of Chemical Engineering, Sungkyunkwan University, Suwon
440-746, Republic of Korea.
Biomineralization is a process which forms a mineral from their precursors
using biomolecules such as peptides or proteins under ambient condition.
Our group previously selected a constrained TiO 2 binding peptide STB1
(CHKKPSKSC) by phage display technique and demonstrated that
STB1 was highly efficient in inducing TiO 2 mineralization at ambient
conditions. In the present study, it is confirmed that kinetics of TiO 2
biomineralization is critical in determining the size, shape and yield of
TiO 2. Fast kinetics favors formation of smaller TiO 2 while slow kinetics
facilitates formation of larger TiO 2. Furthermore, it is found that various
factors, such as peptide concentration, pH, conductivity of mineralizing
m ilieu and presence of salt species, all affect kinetics of TiO 2
biomineralization. Ongoing study will focus on how to leverage the
kinetics-dependent biomineralization phenomenon in view of precisely
controlling the size and morphology of TiO 2. This approach is expected
in near future to provide a new strategy for synthesis of various metal
oxides in controlled, economically viable and environmentally benign
manner.
Keywords : Biomineralization , TiO2(Titanium dioxide) , Kinetics
PFM 11 A ntipro liferativ e A ctiv ities o f Flav o no ids fro m
Vitex rotundifolia
Jung Im LEE, Joo Wan HONG, Myoung Eun JUNG, You Ah KIM 1 and
Youngwan SEO
Division of Marine Environment & Bioscience, Korea Maritime University,
Busan 606-791, Korea.
1
Natural Compounds Bank Establishment Dept., Daegu Gyeongbuk
Institute for Oriental Medicine Industry, Gyeongbuk 712-260, Korea.
The aim of this investigation w as to com pare the antiproliferative
activity of flavonoids (1-3) isolated from the saltmarsh plants Vitex
rotundifolia. The antiproliferative activities against HT1080, AGS, MCF-7
and HT-29 human cancer cells were compared with the control by using
MTT assay. Additionally, expression of major apoptotic genes such as
p21, p53, Bcl-2 and Bax in AGS cells was evaluated by RNA extraction
and RT-PCR m ethod. In the com parative analysis, com pound 2 had
significant antiproliferative activity against AGS with the IC 50 of 6.9 μ
M. It also enhanced expression of tumor suppressor gene p53 and p21 at
a concentration of 25 µM.
Keywords : Vitex rotundifolia , Antiproliferative Activity , Flavonoid
IP: 14.138.203.*** | Accessed 2017/06/02 22:06(KST)
한국생물공학회, 생물공학의 동향(XXIX)：2011. 10
PFM 10 Effects on Antioxidant Activity and Inhibition of
Lactobacillus fermentum by Extracts Isolated from Euonymus
alatus (Thunb.) sieb Leaves and Twigs
Jin Kyu KIM and Ji-young NAM
Korea Atomic Energy Research Institute, Advanced Radiation Technology
Institute, Jeongeup 580-185, Korea.
The Euonymus alatus (Thunb.) sieb. (EA) is a Korean traditional herbal
medicine, commonly used to treat tumors (as a crude drug) in Korea and
China for centuries. The previous studies showed that EA had significant
antidiabetic, antidotic and antiarthritogenic effect, etc. This study was
designed to determine the inhibition effect, antioxidative properties of
crude water-extract(WE), ethanol-extract(EE) and methanol-extract(ME)
from parts of plants from different habitats of EA using isolated
microorganisms from raw rice wine (RRW ). M eanwhile, the Korean
traditional RRW, called Makgeoli, is the most traditional alcoholic
beverage in Korea. However, it deteriorates into bad quality due to a
rapid fermentation. Its short shelf-life is a problem with exporting the
goods to the world market. Treatments of the Lactobacillus fermentum
with various EA extracts changed the bacterial population in the MRS
medium. The results suggested that natural plant extracts such as EA
products could be used as natural preservatives. The results can provide
meaningful data for the RRW to find its way into the competitive world
market.
Keywords : Euonymus alatus (Thunb.) sieb. , Korean traditional raw rice
wine , antioxidative effect
PFM 12 Green Tea Component Theanine Modulates LPS-
Mediated Inflammatory Response in BV2 Microglia
Euteum PARK and Hong Sung CHUN
Dept. of Biotechnology, Chosun University, 375 Seosuk-dong, Gwangju,
Rep. of Korea.
Microglia play a role in the innate immunity in the brain, and over-activated
microglia produce neurotoxic mediators and cytokines, which are inducing
neuronal injury and pathogenesis of the neuroinflammatory diseases.
Recent studies have demonstrated that green tea component L-theanine
has neuroprotective effects and it can easily cross the blood-brain
barrier to act in the brain. How ever, the effect of L-theanine against
the neuroinflammation has not been studied. Therefore, the present
study examined the anti-inflammatory effects of L-theanine against
lipopolysaccharide (LPS)-induced microglial activation. L-theanine
dose-dependently suppressed LPS-induced nitric oxide (NO) release in
BV2 mouse microglial cells. L-theanine also significantly inhibited the
production of TNF-α and IL-1β in LPS-stimulated BV2 cells. In addition,
L-theanine attenuated LPS-induced degradation of IκB-α. Furthermore,
the expression of toll-like receptor 4 (TLR4) was potently inhibited by
L-theanine in BV2 cells. Taken together, L-theanine may act as an anti-
inflammatory agent effectively inhibiting TLR4-mediated microglial
activation.
Keywords : Green tea , Microglia , TLR-4
Recombinant Protein-enabled Biosynthesis of TiOUU₂, WO₃ and TiO₂-WO₃ Hybrid
Metal Oxide Nanoparticles

저자 John MWALE, Noori CHOI, Woo-Seok CHOE

(Authors)

출처 한국생물공학회 학술대회 , 2011.10, 267-267 (1 pages)

(Source)

발행처 한국생물공학회
(Publisher) The Korean Society For Biotechnology And Bioengineering

URL http://www.dbpia.co.kr/Article/NODE02256856

APA Style John MWALE, Noori CHOI, Woo-Seok CHOE (2011). Recombinant Protein-enabled
Biosynthesis of TiOUU₂, WO₃ and TiO₂-WO₃ Hybrid Metal Oxide Nanoparticles. 한국생물
공학회 학술대회, 267-267.

이용정보 14.138.203.***
(Accessed) 2017/06/02 22:07 (KST)

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않습니다. 그리고 DBpia에서 제공되는 저작물은 DBpia와 구독계약을 체결한 기관소속 이용자 혹은 해당 저작물의 개별 구매자
가 비영리적으로만 이용할 수 있습니다. 그러므로 이에 위반하여 DBpia에서 제공되는 저작물을 복제, 전송 등의 방법으로 무단
이용하는 경우 관련 법령에 따라 민, 형사상의 책임을 질 수 있습니다.

Copyright Information
Copyright of all literary works provided by DBpia belongs to the copyright holder(s)and Nurimedia does not guarantee
contents of the literary work or assume responsibility for the same. In addition, the literary works provided by DBpia may
only be used by the users affiliated to the institutions which executed a subscription agreement with DBpia or the
individual purchasers of the literary work(s)for non-commercial purposes. Therefore, any person who illegally uses the
literary works provided by DBpia by means of reproduction or transmission shall assume civil and criminal responsibility
according to applicable laws and regulations.

PEB 49 Biohydrogen production enhancement in [NiFe]-
hy dro g enase ex pressing reco m binant Escherichia coli by
introducing light driven proton pump
Younghwa JO, Jaoon Y.H. KIM, Byung Hoon JO 1 and Hyung Joon CHA
Department of Chemical Engineering, Pohang University of Science
and Technology, Pohang 790-784, Korea.
1
School of Interdisciplinary Bioengineering and Bioscience, Pohang
University of Science and Technology, Pohang 790-784, Korea.
Biohydrogen production can be one of solutions for energy crisis.
Previously, we successfully demonstrated the production of biohydrogen
by constructing [NiFe]-hydrogenase-expressing recombinant E. coli.
Although [NiFe]-hydrogenase showed relatively high oxygen-tolerance
compare to other hydrogenases, still biohydrogen production efficiency
was low. In this research, we introduced proteorhodopsin, light acceptor
for prokaryote working as proton pump, into [NiFe]-hydrogenase-
expressing recombinant E. coli for utilizing light energy for producing
biohydrogen. Proteorhodopsin need a chemical named retinal for working
as proton pump by using light energy but E. coli does not have a
pathway to synthesize it. In order to give E. coli an ability to synthesize
retinal, we expressed five foreign genes expressing enzymes for retinal
synthesis pathway. By introducing light accepting system into E. coli
expressing [NiFe]-hydrogenase, we could get enhanced biohydrogen
productivity.
Keywords : Biohydrogen , [NiFe]-hydrogenase , proteorhodopsin
PEB 51 Improvement of ethanol production from Laminaria
japonica with co-fermentation
Sung-Mok LEE, Geun Hyub KIM and Jae-Hwa LEE
Department of Bioscience and Biotechnology, College of Medical and
Life Science,Silla University, Busan 617-736, Korea.
Ethanol production using Laminaria japonica needs mixed culture of
m icroorganism s for efficient of ferm entation, because of various
carbohydrate. Nuruk contains a mixture of various microorganisms,
including yeast, bacteria, and fungi. Therefore we isolated capable of
ethanol production and hydrolysis microorganisms from Nuruk. The cell
growth and ethanol production of Nuruk strains in terms of fermentation
with different carbon sources (alginate, laminaran, mannitol and L.
japonica) are determined. A maximum cell growth was reached at an
O.D of 4.67 at 600 nm after 72 h for the yeast strain isolated from Nuruk
in the mannitol medium. A high ethanol production was detected on the
mannitol medium, with a highest ethanol production approximately of
2.594 g/L. The overall maximal ethanol production reached was 1.243 g/L
from 20 g/L of L. japonica. A significant increase in ethanol production
was detected with co-fermentation yeast and bacteria isolated from
Nuruk. Maximum ethanol production in the co-fermentation reached
was 2.0 g/L, equivalent approximately 160% of the pure culture.
Keywords : Laminaria japonica , Nuruk , Alginate lyase
IP: 14.138.203.*** | Accessed 2017/06/02 22:07(KST)
한국생물공학회, 생물공학의 동향(XXIX)：2011. 10
PEB 50 Recombinant Protein-enabled Biosynthesis of TiO 2,
WO 3 and TiO 2 -WO 3 Hybrid Metal Oxide Nanoparticles
John MWALE, Noori CHOI and Woo-Seok CHOE
Department of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746.
The use of Genetically Engineered Polypeptides that bind to Inorganics (GEPIs) for in vitro
synthesis of metal oxides such as TiO 2 is emerging as an environmentally friendly alternative
to the harsh reaction conditions normally used in conventional techniques. TiO 2 is commonly
used as a photo catalyst because of its low cost, non-toxicity, chemical stability, and its high
efficiency. However, its large bang gap (~3.2eV) limits its use to the UV light region. By
coupling the two metal oxide semiconductors TiO 2 and WO 3, a reduction in band gap energy
can be achieved, allowing the hybrid to be used in the visible light region. GEPIs are small
peptide sequences selected via combinatorial biology based technologies of phage or cell
surface display. Previously in our group we identified STB1 (CHKKPSKSC), a constrained
heptapeptide cognitive of TiO 2 through a phage surface display technique. By fusing this
peptide to the C-terminus of a DNA binding protein, LacI, we formed a recombinant protein
lacI-STB1. Binding test results showed that it has a high affinity for TiO 2 nanoparticles, as well
as TiO 2, and WO 3 mineralizing ability. A second recombinant protein was obtained by
substituting the lysine residues in STB1 peptide with arginine to form RSTB1 (CHRRPSRSC)
which was then attached to the C-terminus of LacI, resulting in LacI-RSTB1 protein. In the
present study we used LacI-STB1 and LacI-RSTB1 recombinant proteins to induce the
room-temperature formation of TiO 2, WO 3, and hybrid TiO 2-WO 3 nanoparticles. The
synthesized nanoparticles were subsequently quantified and characterized by SEM and EDS.
Compared to using expensive peptides, the use of recombinant proteins, which can be easily
and cheaply obtained via expression in E. coli, presents a less costly route to the biosynthesis
of metal oxides and hybrids for photo catalytic applications.
Keywords : Genetically Engineered Polypeptides for Inorganics , Hybrid metal oxides ,
Photo catalysis
PEB 52 Preparation of Polyester using Waste Liquid Originated
from the Wastepaper Pretreatment by Ethylene Glycol
Dong hun LEE, Dong Il SEO, Chang-Jun KIM and Sung Bae KIM
D epartm ent of Chem ical & Biological Engineering, G yeongsang
National University, Jinju 660-701 Korea.
Polyester from ethylene glycol waste (recycled EG) which was re-used
four times in the pretreatment of wastepaper by ethylene glycol was
prepared. The hydroxyl number of the recycled EG was 441 mg KOH/g
and its composition was cellulose 0.68%, hemicellulose 6.5%, lignin
6.1%, and the rest EG. Maleic acid was used as carboxylic acid and the
effect of carboxyl group/hydroxyl group ratio (C/H ratio), reaction
temperature and time on the crosslinkage of polyester was investigated.
The effect of reaction temperature and time except C/H ratio on the
crosslinkage of polyester was marginal. Citric acid, lithium hydroxide
or dicumyl peroxide was used as additive to enhance the crosslinkage of
polyester, and among them, 10% citric acid was found to be most
effective. The crosslinkage was 86% when the polyester was prepared
at an optimum condition such as 130℃ and 15 minutes of reaction
condition, 1.5 of C/H ratio, and 10% citric acid, and its insoluble
percent in boiling water for 6 hours was 47%. The weight loss in 5
months was approximately 40%, indicating that the prepared polyester
is readily biodegradable.
Keywords : polyester , ethylene glycol , pretreatment , wastepaper ,
crosslinkage