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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal DOI :
Korean Society for Biotechnology and Bioengineering
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Volume & Issues
Volume 21, Issue 6 - Dec 2006
Volume 21, Issue 5 - Oct 2006
Volume 21, Issue 4 - Aug 2006
Volume 21, Issue 3 - Jun 2006
Volume 21, Issue 2 - Apr 2006
Volume 21, Issue 1 - Feb 2006
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Informatics Network Representation Between Cells Using Probabilistic Graphical Models
Ra, Sang-Dong ; Shin, Hyun-Jae ; Cha, Wol-Suk ;
KSBB Journal, volume 21, issue 4, 2006, Pages 231~235
This study is a numerical representative modeling analysis for the application of the process that unravels networks between cells in genetics to web of informatics. Using the probabilistic graphical model, the insight from the data describing biological networks is used for making a probabilistic function. Rather than a complex network of cells, we reconstruct a simple lower-stage model and show a genetic representation level from the genetic based network logic. We made probabilistic graphical models from genetic data and extends them to genetic representation data in the method of network modeling in informatics
The Dependency of the Expression Level of Recombinant Protein by the Drop of Alkali Consumption Rate after Induction
Hur, Won ;
KSBB Journal, volume 21, issue 4, 2006, Pages 236~240
IPTG induction caused a sudden drop of alkali consumption rate during cultivation of a recombinant E. coli with
-galactosidase structural gene under T7 promoter on a plasmid. A series of batch cultivations showed the positive correlation of the decrease of alkali consumption and the level of expression. However, repeated IPTG induction did not cause any variation of alkali consumption rate. Supplementation of medium even at stationary phase enhanced the level of
-galactosidase expression. These results suggests that the drop of alkali consumption rate by IPTG induction represents the rate of expression.
On-Line Measurement of Biochemical Oxygen Demand of livestock Wastewater by Multi-Biosensor System
Kim, Jin-Kyeung ; Kim, Tai-Jin ;
KSBB Journal, volume 21, issue 4, 2006, Pages 241~247
The present study was intended to examine a basic scheme to determine the biochemical oxygen demand(BOD) of livestock wastewater by means of six individual dissolved oxygen(DO) sensors and its multi-measurable meter. Maximal point of the first order time derivative of the DO difference between DO distribution of sterilized livestock wastewater and that of non-sterilized livestock wastewater, was considered as the oxygen uptake rate(OUR) of microorganisms in livestock wastewater, as determined to be 0.00074 mg
. The present study showed that there was a fair linear relationship(97.72%) between maximal OUR and BOD values of livestock wastewater, the latter being determined by classical Winkler azide method. It was thus concluded that the present multi-biosensor system might be applicable to an on-line system for measurement of BOD of livestock wastewater.
Improvement of Proliferation Capacity of Non-adapted CHO Cells Subcultured Using Serum Free Media in Long-term Culture
Lee, Seung-Sun ; Lee, Jin-Sung ; Byun, Soon-Hyu ; Park, Hong-Woo ; Choe, Tae-Boo ;
KSBB Journal, volume 21, issue 4, 2006, Pages 248~254
Animal cell culture industry has a large market and an exponential growth rate among biological industry field. Chines hamster ovary(CHO) cells are the most widely used cell lines for recombinant protein production. They can avoid infection from polio, herpes, hepatitis B, HIV, measles, adenovirus and etc. Moreover it is easy to transfection recombinant genes and possible to suspension culture. Serum free media is one of the most important factor of protein production. Because serum has problems. Serum is not defined the contents until now, it has a number of proteins, lipids, carbohydrates and unknown molecules that cause of risk involve in infection and high cost of product purification. CHO cell line cultured using serum free media were the basis of a very successful method to produce(glyco-)protein in mammalian cells, which are then used as pharmaceutical products. Also, the low protein content of the developed medium facilitates downstream processing and product purification. But non-adapted CHO cells have a limit of proliferation cultured using serum free media and it takes very long time to adapt non-adapted cells to serum free media. There are a number of causes of a limit of proliferation using serum free media. Absence of growth factors and growth stimulating molecules is a major factor of the reasons. It makes growth signals and moves cell cycle. And increase of cellular stress is another reason. It induces increase of intraceullar ROS concentration. The purpose of this study is about improvement of proliferation capacity of non-adapted CHO cells cultured using serum free media without adaptation process.
Cytotoxicity of Hyaluronic Acid Membrane Cross-linked with Lactide
Kim, Won-Jung ; Kwon, Ji-Young ; Cheong, Seong-Ihl ; Kim, In-Seop ;
KSBB Journal, volume 21, issue 4, 2006, Pages 255~259
The biodegradable hyaluronic acid(HA) membranes cross-linked with lactide using the crosslinking agent, 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide(EDC) were prepared as a potential biocompatible material for tissue engineering. HA membranes having different mechanical properties were synthesised by varying degree of the mole ratio of lactide to HA, EDC concentration, and crosslinking temperature. HA membranes were degradable in water solution and the degradation became slower with the increasing mole ratio of lactide to HA. HA membranes were sterilized using ethylene oxide gas and extracted with cell culture medium for 24 h at
and 200 rpm. Cytotoxicity of the extract was tested using NIH/3T3 mouse embryo fibroblast as a model cell. Growth inhibition was not observed in the extracts of HA membranes with the mole ratios of lactide to HA, 5 or 10, and 10% EDC concentration, however 11% of growth inhibition was observed in the extract with the mole ratio of 13. Growth inhibition was not observed in the extracts of HA membranes prepared with 5% EDC or 10% EDC and the mole ratio of lactide to HA, 10, however 12% of growth inhibition was observed in the extract with 20% EDC. Cytotoxicity was not observed in the extracts of HA membranes prepared at varying crosslinking temperatures,
with the mole ratio of lactide to HA, 10 and 10% EDC.
Effect of Various Additives on the Production of Recombinant HBsAg during Methanol Induction in Pichia pastoris
Lee, Kyoung-Hoon ; Lim, Sang-Min ; Kim, Dong-Il ;
KSBB Journal, volume 21, issue 4, 2006, Pages 260~266
Methanol induction conditions with various additives for the enhanced production of recombinant hepatitis B surface antigen(HBsAg) were investigated in Pichia pastoris, which can utilize methanol as a carbon source and produce recombinant proteins under the control of strong, tightly-regulated alcohol oxidase(AOX) promoter. The presence of non-methanol carbon sources such as glycerol and glucose fully repressed the expression of AOX promoter. Various additives were tested to improve the production of recombinant protein and it was found that sorbitol could be a good carbon source during methanol induction period. An optimized concentration of amino acid mixture enhanced the production of HBsAg significantly. Pluronic F-68, a non-ionic surfactant, also improved the production of HBsAg without inhibiting cell growth. Addition of oleic acid at 0.01%(v/v) during the induction period showed positive effect on the production of HBsAg. Finally, 1.2%(v/v) of trace salts enhanced the production of HBsAg 1.9 times compared to that of control culture.
Isolation and Characterization of a Strain for Economical Ethanol Production
Han, Hyo-Jung ; Kim, Seong-Jun ;
KSBB Journal, volume 21, issue 4, 2006, Pages 267~272
Five strains producing ethanol were isolated from soil near traditional alcohol production factory in Gwangju, Korea. One of the isolated strains maintained relatively stable ethanol production in shaking culture. The isolated strain KJ was proved to be Saccharomyces italicus, based on several biochemical and morphological tests containing assimilation of carbon compounds. In investment of the most suitable carbon for ethanol production, ethanol concentration of 5.46 g/L and yield of 0.53 g-ethanol/g-glucose were obtained in condition of glucose 10 g/L in YM medium. Experimental optimal conditions for ethanol fermentation by S. italicus KJ were as follows; temperature
, initial pH 5.0, initial concentration 10% of glucose, anaerobic condition in the liquid cultivation. When enzymatically saccharified food wastes(SFW) were used as the production medium, ethanol production yield was 0.57 g-ethanol/g-reducing sugar. Therefore, SFW will contribute to lower the production cost of ethanol for industrial application.
Microcontact Printing of Bacteria Using Hybrid Agarose Gel Stamp
Shim, Hyun-Woo ; Lee, Ji-Hye ; Lee, Chang-Soo ;
KSBB Journal, volume 21, issue 4, 2006, Pages 273~278
The noble method of hybrid agarose gel microstamp fabricated by replica molding against PDMS master to make bacteria patterns on agar surface was presented. After the fabricated hybrid agarose gel microstamp was inked with E. coli, we could obtain 2 dimensional bacterial arrays with
circular spots. And the various shaped patterns based on experimental design were easily generated. The analysis of mean fluorescent signal was showed that bacterial pattern have high contrast between spots and background and homogeneity of pattern. Our proposed method solved the problem of wetting and handling with small soft agarose gel microstamp when bacteria were used for ink. The agarose gel stamp provides appropriate environment to inked bacteria, which is essential technology for cell patterning with high retaining viability during the patterning process. This method is reproducible, convenient, rapid, and could be applied to screening system, study of cell-surface interaction, and microbial ecology.
Comparison of Enzymatic Activity and Cleavage Characteristics of Trypsin Immobilized by Covalent Conjugation and Affinity Interaction
Jang, Dae-Ho ; Seong, Gi-Hun ; Lee, Eun-Kyu ;
KSBB Journal, volume 21, issue 4, 2006, Pages 279~285
We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized
mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was
, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.
Fermentation Kinetics for Production of Carotenoids by
-ionone Resistant Mutant of Xanthophyllomyces dendrorhous
Park, Ki-Moon ; Kim, Young-Jun ; Song, Min-Woo ; Kang, Seog-Jin ; Lee, Jae-Heung ;
KSBB Journal, volume 21, issue 4, 2006, Pages 286~291
-ionone resistant mutants were isolated from the wild-type red yeast Xanthophyllomyces dendrorhous KCTC 7704. Although the growth of X. dendrorhous KCTC 7704 was strongly inhibited at 0.025 mM
-ionone, one of the
-ionone resistant mutants isolated at 0.1 mM
-ionone by NTG mutagenesis showed rather 70% of relative survival at 0.15 mM
-ionone. Fermentation kinetics study with the mutant was carried out at
for 4 days in 300-mL baffled flasks. The mutant yielded up to 2.3-fold higher carotenoids content(viz.
of total carotenoids per mg of dry cells) compared with the wild-type strain. The production of metabolites such as organic acids could be neglected. Studies on the kinetics with various carbon substrates revealed both an increase in final dry cell mass and a higher total carotenoids content in cell mass with glucose when compared to fructose or sucrose. As a further part of study, the effect of pH on the fermentation kinetics was investigated in glucose-limited chemostat at a dilution rate of
. When compared to steady-state kinetic parameters obtained at pH 4.0, a significant reduction in cell concentration at pH 3.0 and a lower carotenoids content at pH 5.2 were evident.
Effect of Phellinus Extracts on Sprouting in Porcine Pulmonary Artery Endothelial Cells
Oh, In-Suk ; Kim, Hwan-Gyu ;
KSBB Journal, volume 21, issue 4, 2006, Pages 292~297
One of the steps in angiogenesis is the degradation of the underlying basement membrane via proteases. Endothelial cells release proteinases to degrade the extracellular matrix for their sprouting in vivo. In this study, we examined the effect of water extracts of Phellinus linteusis(Phellinus extracts) and combination of Phellinus extracts and fibroblast growth factor(FGF-2) on cultured porcine pulmonary artery endothelial cells(PPAECs). Phellinus extracts induced sprouting of PPAECs, which was inhibited by MMPs and plasmin inhibitors, and induced the secretion of matrix metalloproteinase-3(MMP-3) and plasmin. At high concentration of Phellinus extracts(
), the active MMP-2 secretion was induced. It is therefore, suggested that Phellinus extracts induces the sprouting of cultured endothelial cells by means of increased active MMP-2 and plasmin secretion. Also, combination with Phellinus extracts and FGF-2 produced an enhanced effect on sprouting and secretion of active MMP-2, and MMP-3 and plasmin from PPAECs.
Production of Recombinant Trehalose Synthase from Thermus caldophilus GK24
Choi, Jae-Youl ; Cha, Wol-Suk ; Shin, Hyun-Jae ;
KSBB Journal, volume 21, issue 4, 2006, Pages 298~301
A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.
Rapid Preparation of Truncated Transaminases using a PCR-based Cell-free Protein Synthesis System
Kwon, Yong-Chan ; Park, Kyung-Moon ; Kim, Dong-Myung ;
KSBB Journal, volume 21, issue 4, 2006, Pages 302~305
In this work, we attempted the application of cell-free protein synthesis technology for the rapid generation of truncated enzymes. Truncated DNAs of a transaminase were PCR-amplified and directly expressed in cell-free protein synthesis reactions. Variants of the transaminase were rapidly prepared and analyzed for their enzymatic activity. Described method that combines the PCR and cell-free protein synthesis technologies will offer a versatile platform for the rapid generation of optimally modified protein species.
Short-term Hypothermic Preservation of CHO Cells Using Serum-Free Media
Byoun, Soon-Hwi ; Park, Hong-Woo ; Choe, Tae-Boo ;
KSBB Journal, volume 21, issue 4, 2006, Pages 306~311
Cell preservation is indispensable in animal cell culture process and should be established according to the cell characteristics. In this study, we experimented hypothermic preservation of CHO cells that is widely used in pharmaceutical industry to produce therapeutic proteins and established a stable method of preservation. The highest viability of CHO cells was obtained when the cells were preserved using rolling tube, which means the cells should be suspended to avoid the cell lumping during the preservation. Also, we obtained superior preservation result under the anaerobic condition. To evaluate the serum-free media as a preservation solution, we investigated cell growth after hypothermic preservation using serum-free media. High cell viability and normal cell growth was observed during 10 days using serum-free media. Moreover, we found that more effective preservation when
-tocopherol and retinoic acid is added to media as an additive. In the case of 1 liter large scale hypothermic preservation using established protocol, cell viability and growth rate was obtained as good as small scale one. This study is considered to be helpful for hypothermic preservation of CHO cells and large scale hypothermic preservation may be available through the further studies.
A Study on the Antimicrobial Effect of Ginkgo biloba Leaves Extracts according to Concentrations of Ethanol for staphylococcus aureus
Lee, In-Hwa ; Shim, Youn ; Choi, Seung-Hyun ; Park, Ju-Young ; Han, Sung-Woo ; Song, Jn-Young ; Yoon, Suk-Jin ;
KSBB Journal, volume 21, issue 4, 2006, Pages 312~316
A optimal condition for the Ginkgo biloba extraction in ethanol and water binary solvent system has been proposed based on concentration of bilobalide and ginkgolide known as having a antimicrobial components in the range 5% to 70% ethanol in water at
. Concentration of bilobalide as a single component of Ginkgo biloba leaves extract is the highest at the 60% ethanol and ginkgolide A and B is highest at 50% ethanol. Antimicrobial effect of Ginkgo biloba leaves extracts on the S. aureus was also examined by disc diffusion test and optical density test. In case of the disc diffusion test, the clean zone diameter was increased from 0.95 cm to 1.70 cm as ethanol concentration increased from 5 to 70%. However, over the 40% of ethanol concentration the antimicrobial effect was almost flat. Based on these results, we propose that the 40% of ethanol and 60% water solvent is most desirable for Ginkgo biloba extract considering vapor pressure problem in concentrating process after extraction. We introduced SEM and TEM to figure out the morphological change on the surface and inside body of S. aureus when Ginkgo biloba leaves extract was treated. After mixed with Ginkgo biloba leaves extract blast like blebs appeared on the surface of S. aureus cells and cell wall was not observed. From the these results, it seems that the Ginkgo biloba leaves extract including bilobalide and ginkgolide A, B prevent cell wall synthesis.
A Study on the Utilization of Effluent Treated by Double Process Using Fixed-media and Sand Filter Coated by Nano Silver for Wastewater Reclamation and Reusing System
Seon, Yong-Ho ;
KSBB Journal, volume 21, issue 4, 2006, Pages 317~323
This study focused on the availability of wastewater reclamation and reusing system as one of the alternatives against the global water shortage in near future, which system is composed of two treatment steps; first, wastewater is injected into upflow
biofilm process(anaerobic/anoxic/oxic) reactor filled with polyethylene fixed-media, and the effluent of 1st steps continuously passed through downflow nano silver sand filter. The pH of the effluent ranged from 7.39 to 8.06(average 7.84), the
(average 12.1 mg/L), and
(average 4.9 mg/L), that met all the wastewater reclamation and reusing system criteria. Besides, the SS concentrations of the effluent which was
(average 4.95 mg/L) met the criteria(5 mg/L), showing 94.8% of average removal efficiency. The 99.1% of the average removal efficiency of the E-coliform did not met the criteria(Not detected), which indicates the needs for the following chlorine disinfection treatment with the residual chlorine concentration of above 0.2 mg/L. There are no bacteria on the sand surface coated by nano silver. The removal efficiency of T-N and T-P that could be included into the criteria in the future was 50.3% and 27.2% respectively.