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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 18, Issue 6 - Dec 2003
Volume 18, Issue 5 - Oct 2003
Volume 18, Issue 4 - Aug 2003
Volume 18, Issue 3 - Jun 2003
Volume 18, Issue 2 - Apr 2003
Volume 18, Issue 1 - Feb 2003
Selecting the target year
Two-Stage Fed-Batch Culture of Candida magnoliae for the Production of Erythritol using an Industrial Medium
KSBB Journal, volume 18, issue 4, 2003, Pages 249~254
Experiments were carried out to select an industrial nitrogen source and optimize erythritol production by Candida magnoliae in fed-batch culture. Among the industrial nitrogen sources tested, light steep water (LSW) was found to be the best nitrogen source for producing erythritol, based on erythritol yield and raw material price. The maximum erythritol concentration obtained a 131.6 g/L, with a 52.6％ yield and 0.52 g/L-hr productivity from a 250 g/L glucose and 43.3 mL/L LSW in batch culture. Two-stage fed-batch culture was chosen to improve the volumetric productivity and the yield of erythritol. High cell density culture in cell growth stage was achieved by batch type culture containing 100 g/L glucose and 500 mL/L LSW. The cell concentration was 71.0 g/L after 23 hours of culture. Erythritol productivity was decreased by increasing glucose concentration in the production stage. But 37.3％ of the maximum erythritol yield was obtained with 185.5 g/L of erythritol and 1.66 g/L-hr of productivity when 820 g of glucose powder was directly added to a concentration of 450 g/L glucose in production stage.
Replacement of the in vivo Bioassay for Erythropoietin with the in vitro Bioassay
KSBB Journal, volume 18, issue 4, 2003, Pages 255~260
In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(％) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 ％ ＝0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-
, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-
) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-
with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.
Empirical Equation for Resolution if Ibuprofen Enantiomers by Chiral High-Performance Liquid Chromatography
KSBB Journal, volume 18, issue 4, 2003, Pages 261~265
Ibuprofen was analyzed by chiral high performance liquid chromatography. Retention behaviours of the standard mixtures of ibuprofen were investigated to obtain their acceptable resolution. A chromatographic column (3.9
300 mm) was packed by Kromasil CHI-TBB packings (10
) and n-hexane was used as a mobile phase with 0.1％ acetic acid and tert-butyl methyl ether. Isocratic elution of ibuprofen at 1.0
/min was performed by changing the mobile phase compositions. The experimental variables affecting the resolution were the compositions of mobile phase and chemical buffer (n-hexane and tert-butyl methyl ether). The resolution between the enantiomers were correlated into the several types of empirical equations including linear form, and their agreements between experimental data and calculated values were examined by the regression coefficient.
Antioxidant Activities of Stachys sieboldii MIQ. Stalks
KSBB Journal, volume 18, issue 4, 2003, Pages 266~271
This study was worked out to investigate the compounds of antioxidant constituents extracted from Stachys Sieboldii MiQ. and their effects on antioxidant activity by DPPH method, ferric thiocyanate method, and nitrite scavenging ability. Solvents such as methanol, hexane, chloroform, ethyl acetate, butanol, and water were used for this purpose. Total concentrations of polyphenols and flavonoids were measured in the methanol fraction. The ethyl acetate fraction showed the strongest activity by DPPH method, ferric thiocyanate method, and nitrite scavenging ability. The ethyl acetate extract was fractionated on a silica gel column using elution solvent (chloroform: methanol: water ＝ 70 : 30 : 5 lower phase) at a flow rate of 1.0 mL/min. UV-VIS spectral data of each fraction showed adsorption maxima in the range of 284～330 nm. Among fractions, the fraction 1 that has λ
max/ (nm) of 284 nm showed the strongest activity by DPPH method. The UV-VIS spectral data of phenolic compounds were known to lie in the range of 210～290 nm and 300～550 nm. Therefore, the results of our study suggested that Stachys sieboldii MIQ. contains phenolic compounds showing natural antioxidant activity.
Treatment of Food Processing Wastewater bearing Furfural by Candida utilis
KSBB Journal, volume 18, issue 4, 2003, Pages 272~276
A yeast treatment process was applied to treat food processing organic wastewater containing inhibitory material to anaerobic bacteria. The wastewater contained high concentration of the furfural as a by-product from the food processing. Aerobic yeast (Candida utilis) was selected to remove organics in wastewater. The batch test showed that the wastewater had an inhibition to anaerobic bacteria. The optimum level of temperature for yeast treatment was ranged from 25 to 45
. The pH range from 4 to 8 was favorable to yeast growth. The continuous flow reactor was operated at various SRTs. The results were satisfactory with the reduction of COD up to 90％ at SRT of more than 1 day. Through the kinetic study of the yeast, the remained COD concentration was mainly caused by the formation of soluble microbial product (SMP).
The Growth Characteristics of Spirulina platensis in Cylindrical Photobioreactor
KSBB Journal, volume 18, issue 4, 2003, Pages 277~281
The study of growth characteristics for Spirulina platensis were carried out in 400 mL cylindrical photobioreactor and the effects of carbon dioxide concentration and flow rate during the growth of Spirulina platensis were investigated. The results showed that relatively low carbon dioxide concentration and high flow rate forced the growth of Spirulina platensis in experiment conditions. The pH analysis showed that different carbon dioxide concentration might form particular aqueous carbonate system in culture medium and affect the growth of Spirulina platensis. In addition, the possibility of limiting light radiation by cell density was investigated by the analysis of specific growth rate. The result intimated that the cause of decrease of specific growth rate at exponential phase was due to the limitation of light radiation by Spirulina platensis cell density in cylindrical photobioreactor.
Screening of Hyaluronidase Inhibitory and Free Radical Scavenging Activity in vitro of Traditional Herbal Medicine Extracts
KSBB Journal, volume 18, issue 4, 2003, Pages 282~288
For the screening of anti-inflammation and antioxidative activities, ethanolic extract of 40 species of traditional herbal medicines were examined their hyaluronidase inhibitory effect and radical scavenging activity in vitro. From the result of the hyaluronidase inhibitory activity using a Morgan-Elson assay, Astragali Radix, Eucommia Cortex, Schizandrae Fructus, Scutellaria Radix, Acanthopanacis Cortex, Chaenomelis Fructus, Amomum xanthioides Wallich and Moutan Radicis Cortex showed more than 50% hyaluronidase inhibitory effects at the concentration of 10 mg/mL. In the various solvent fractions (n-hexane, chloroform, ethyl acetate, water) prepared from ethanolic extracts, the ethyl acetate fraction of all extracts tested showed strong activity. Antioxidative activity was evaluated by assaying electron-donating ability to DPPH free radical and scavenging of hydroxyl radical (ㆍOH) generated through Fenton reaction, respectively. Rubus coreanus Miq, Moutan Radicis Cortex, Paeoniae Radix Alba, Plantaginis Semen and Sorbus commixta Hedl. showed high activity more than 90%, yet similar activity to
-tocopherol and BHA at the concentration of 1 mg/mL in electron donating activity. The scavenging effects of ethanolic extracts on hydroxyl radical were investigated using a 2-deoxyribose oxidation method and tested all extracts showed significant radical scavenging activity. The experiment was also performed to examine whether herbal medicines having significant lipid peroxidation inhibitory activity, Schizandrae-Fructus is the strongest inhibitory activity in both linoleic acid and liposome peroxidation.
Development of Doxorubicin overproducing Streptomyces Strain using Protoplast Regeneration
KSBB Journal, volume 18, issue 4, 2003, Pages 289~293
Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. In order to increase doxurubicin productivity, a semi-industrial doxorubicin-producing Streptomyces strain named BR-Dox was cultured in a R2YE liquid medium containing CaCO
, and then converted to a cell wall-free protoplast using lysozyme treatment method, followed by PEG-mediated cell wall regeneration. Among several protoplast-regenerated Streptomyces BR-Dox strains, two independent isolates named BR-Dox4 and BR-Dox6 were visually selected using thin layer chromatography (TLC) based on the pigment overproducing phenotype. Comparing with Streptomyces BR-Dox parental strain, two protoplast-regenerated strains, BR-Dox4 and BR-Dox6 exhibited 25.2％ and 12.2％ higher doxorubicin productivity analyzed by high pressure liquid chromatography (HPLC), respectively. This result suggests that a protoplast-regeneration of an antibiotics-producing Streptomyces strain should be a promising strain development approach for antibiotics overproduction in Streptomyces species.
Improved Viability and Proteome Analysis of Lactobacillus fermentum KLB12 upon Heat Stress
KSBB Journal, volume 18, issue 4, 2003, Pages 294~300
In the previous study, we have isolated several vaginal lactobacilli from Korean woman and selected one of them (KLB12) for further study, which was indentified as Lactobacillus fermentum by sequence analysis of 16S rRNA gene. Formulated L. fermentum KLB12 can be used for ecological treatment of bacterial vaginosis. For pharmaceutical formulation, the spray-drying process is required where stress such as high temperature is routinely applied. In this study, we found that heat stress at 60
for 20∼30min reduced the viable cell population of KLB12 by 10
9/. However, adaptation of KLB12 cells at 52
made them more thermotolerant upon exposure to 60
. The level of thermal protection in RSM (reconstituted skim milk) by adaptation in acid (pH 5), cold (4
), ethanol (3％), NaCI (0.3M) was also examined. Although not as efficient as the homologous stress, adaptations in both cold and NaCI gave considerable cross protection against heat stress. When chloramphenicol was added during heat adaptation, adaptation effect was abolished. This suggests that de novo protein synthesis is necessary during the adaptation process. Important changes in proteome during heat adaptation was examined with two-dimensional gel electrophoresis.
Pancreatin Production by Removal of Lipid from Hog Pancreas using Supercritical Carbon Dioxide with Entrainer
KSBB Journal, volume 18, issue 4, 2003, Pages 301~305
The study of pancreatin extraction was investigated by supercritical fluid process. Using supercritical carbon dioxide extraction with entrainer the purification of pancreatin was possible to remove lipids from Hog pancreas. To observe the optimum conditions different experimental variables were changed as pressure, temperature, flow rate of solvent and 0.25 mm of sample size were evaluated for effective removal of lipids. Ethanol and n-hexane were used as an entrainer with 5 mL/min. Increasing pressure at constant temperature the efficiency of the lipid removal in Hog pancreas was improved and the protein was concentrated without denaturalization, compared that of the control Hog pancreas. The most efficient conditions of lipid elimination were 17 MPa of pressure and 35
of temperature and 0.25 mm of sample size.
Solid-phase Refolding of Immobilized Enterokinase for Fusion Protein Cleavage
KSBB Journal, volume 18, issue 4, 2003, Pages 306~311
Solid-phase refolding of immobilized proteins can be an effective way to reuse an immobilized enzyme column. Oriented immobilization methods are known to provide higher activity of the immobilized enzymes. In this study, using recombinant EK (enterokinase) as a model enzyme and a fusion protein, that consisted of recombinant human growth hormone and six His tag that was linked by the peptide of EK-specific recognition sequence, as a model substrate, we evaluated two oriented immobilization methods, i. e., reductive alkylation of N-terminus
-amine and affinity interaction between poly-histidine tag and Ni-NTA (nickel-nitrilotriacetic acid). The immobilization yield, activity and cleavage of the immobilized enzymes, and the yield of solid-phase refolding were compared. The Ni affinity immobilization and the covalent immobilization yields were about 100％ and 65％, respectively. But the specific activities were the same, about 50％ of that of the soluble enzyme. The cleavage rate by the covalently immobilized EK was higher than the soluble enzyme and the side reaction of cryptic cleavage was significantly decreased. Covalently immobilized EK showed almost 100％ refolding yield but the affinity immobilized EK showed only 70％ yield, which suggested the covalent conjugation provided more rigid ‘reference structure’ for the solid-phase refolding. The monomeric hGH could be easily obtained by capturing the cleaved poly Histidine tag by the Ni affinity column.
Effect of Glycine Supplement on Extracellular Secretion of Levansucrase form Pseudomonas aurantiaca S-4380 in Recombinant Escherichia coli
KSBB Journal, volume 18, issue 4, 2003, Pages 312~317
The addition of glycine up to 0.5％ (w/v) to Luria broth (LB) media on the secretion of levansucrase in a recombinant strain Escherichia coli JM109/pUPLK1 was observed to enhance the release of periplasmic proteins from the cell to the broth, without significantly affecting the cell growth rate and protein productivity. However, the glycine concentration at 1 ％ (w/v), the cell density attainable at the stationary phase fell to about 50％ and the extracellular activity of levansucrase corresponded to about 80％ of the total (extracellular plus intracellular) activity and increased by 2.6-fold, comparing to the cells grown in the absence of glycine. The increased pH at stationary phase accelerated the degradation of levansucrase. Maximal extracellular activity was attained when 1 ％ glycine was supplemented at the onset of strain growth.
Anaerobic Acid and Methane Fermentation using Paper Wastes
KSBB Journal, volume 18, issue 4, 2003, Pages 318~323
This study was carried out to investigate the possibility of reusing newspaper or paperbox waste by methane fermentation. When 15 g of newspaper and box wastes were digested separately for 24 days by batch fermentation, the amount of total organic acids produced were 2461 and 4978 mg/L, respectively. The tCOD removal rates were found to be 60.9 and 62.4%, respectively. In addition, the removal efficiencies of total solid were 34.8 and 33.4%, and those of volatile solid were 40.0 and 39.2%, respectively. During this period, the amounts of biogas produced were 6.95 and 6.43 L. In a semicontinuous reaction, tCOD removal efficiencies for newspaper and box wastes were 64.7 and 65.0%, respectively, after 14 days of digestion. After 25 days, which were needed to stabilize the methane fermentation, the amounts of biogas produced daily were 0.31 and 0.30 L/g.dry wt, respectively. Methane contents were 57.3 and 56.2%, respectively, and the pHs in the anaerobic acidogenic and methanogenic fermenters were 5.0 and 7.5, respectively.
Isolation and Characterization of Antitumor Agents from Xanthium strumarium L.
KSBB Journal, volume 18, issue 4, 2003, Pages 324~328
In a mutagenicity test using the Salmonella typhimurium TA98 and TA100, the Xanthium strumarium L. extracts had not a mutagenicity. The extracts were assayed that antioxidative effect using a colony formation assay. The extracts showed protective effects against the cytotoxicity of H
and increased the immunity induced by TNF and IL-1
. The modulating effect of Xanthium strumarium L. extract on the induction of carcinogenesis by N-methyl-N'-nitro-N-nitrosoguanidin (MNNG), was investigated in Wistar rats. The GSH content was found to be reduced by MNNG treatment, but increased on adding extract. In addition the Xanthium strumarium L. extract increased p53 expression versus MNNG alone.
Regulatory Mechanism in Tissue-specific Expression of Insulin-like Growth Factor-I Gene
KSBB Journal, volume 18, issue 4, 2003, Pages 329~334
The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBP
proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42
/, and p35
form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42
/ and p38
/ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.
Effect and Safety of Replacement Therapy with Insumon in Type 2 Diabetes Mellitus
KSBB Journal, volume 18, issue 4, 2003, Pages 335~339
This research was concentrated to Investigate the effects of Insumon as a newly developed dietary supplement in patients with type 2 diabetes. The average glycated hemoglobin level was dropped from 7.0
1.1％ to 6.7
0.9％ after 8 weeks with statistic significance (p＝0.0202). For patients of HbA1c over 7％, the average HbA1c significantly decreased from 8.1
0.8％ to 7.5
0.8％ (p＝0.0171). The average fasting blood glucose level was decreased significantly from 9.5
1.2 mmol/L (171.6
22.3 mg/dL) to 8.5
1.5 mmol/L (152.3
26.3 mg/dL) (p=0.0262) after 4 weeks and to 8.8
1.5 mmol/L (158.3
24.1 mg/dL) (p=0.0445) after 8 weeks in the group of patients of FBG of over 7.8 mmol/L (140 mg/dL). The average HDL cholesterol level was increased significantly after 8 weeks from 1.2
0.2 mmol/L (46.5
8.6 mg/dL) to 1.3
0.3 mmol/L (51.6
10.9 mg/dL) (p=0.0007). Thus, these results suggested that patients with type 2 diabetes might benefit by Insumon as a herbal dietary supplement without any serious side effects.
Effect of Light on Growth of Chlorella sp. HA-1 in Large-scale Photobioreactors for Biological
KSBB Journal, volume 18, issue 4, 2003, Pages 340~345
Feasibility of large-scale photobioreactors for biological CO
fixation was investigated using Chlorella sp. HA-1. Generally, as the volume of photobioreactor increased, the CO
fixation rate decreased because of a lower illumination efficiency in large-scale than in small-scale photobioreactors. Though controlling the arrangement and the number of light source, the maximum CO
fixation rates that could be achieved were 530 and 357 gCO
/㎡day for 40 L and 188 L photobioreactor, respectively, which was higher than the CO
fixation rate of lab-scale photobioreactor.