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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 13, Issue 6 - Dec 1998
Volume 13, Issue 5 - Oct 1998
Volume 13, Issue 4 - Aug 1998
Volume 13, Issue 3 - Jun 1998
Volume 13, Issue 2 - Apr 1998
Volume 13, Issue 1 - Feb 1998
Selecting the target year
Purification and Properties of
KSBB Journal, volume 13, issue 1, 1998, Pages 1~5
-fructofuranosidase of garlic (Allium sativum L. ; Seosan) was purified by ammonium sulfate fractionation and gel filtration chromatography with a recovery of 8.3%. The molecular mass of the purified enzyme was estimated to be 79kDa by SDS-PAGE, which revealed two subunits of 41kDa and 38kDa. Maximum enzyme activity was observed at pH 5.0 and 40
and the enzyme was stable over the pH range of 4.0-6.0 and below 50
. The Km value for sucrose was 15.5mM and the enzyme activity was significantly inhibited by bivalent metal ions(Hg
) and EDTA.
Production of Xylitol by Catabolite Derepressed Mutant of Candida sp.
KSBB Journal, volume 13, issue 1, 1998, Pages 6~12
In order to produce xylitol from hemicellulose hydrolysate which is widely used as a substrate, the development of strain such as catabolite derepressed mutant is required. After treatment of Candida sp. with EMS, GM-17 and PM-34 as catabolite derepressed mutant were isolated from Candida guilliermondii and Candida parapsilosis, respectively. Mutant GM-17 and PM-34 simultaneously assimilated xylose and glucose during the fermentation. The specific xylose reductase and xylitol dehydrogenase activities of mutant strains were also higher than those of wild strains in glucose medium and mixed medium of glucose and xylose. The xylitol productivity and yield of mutant GM-17 and PM-34 were improved as compared to the wild types in the mixed medium. The xylitol productivity and yield of mutant GM-17 were 0.09 g/L·hr and 0.56 g-xylitol/g-xylose, and those of mutant PM-34 were 0.21 g/L·hr and 0.52 g-xylitol/g-xylose in the mixed medium, respectively.
Animal Cell Culture and the Production of Monoclonal Antibody(MAb) Using Biopolymer Membrane
KSBB Journal, volume 13, issue 1, 1998, Pages 13~19
Biopolymer membrane was prepared using two oppositely charged natural biopolymers. The biopolymer membrane was used for the encapsulation of two hybridoma cell lines(ATCC CRL-1606, ATCC HB-8852) to produce monoclonal antibodies. In order to reduce the down stream steps, the pre size of the membrane was controlled to retain the monoclonal antibodies in the capsules based on the diffusion experiments with standard proteins. T-flask culture showed cell densities of 8
107 cells/mL and 3
107 cells/mL, and MAb concentrations of 506
g/mL and 109
g/mL for encapsulated ATCC CRL-1606 and HB-8852, respectively. Two liter perfusion cultures with encapsulated ATCC HB-8852 were performed to enhance the MAb production. The MAb production of the encapsulated hybridoma increased considerably comparing to the culture using silicon tubing for oxygen transfer.
Optimum Conditions of Cellulose-Hydrolysis Reaction with Mixed Enzymes of Cellulase and
KSBB Journal, volume 13, issue 1, 1998, Pages 20~25
Optimum conditions of the cellulose-hydrolysis reaction with mixed enzymes(cellulase extracted from Penicellium funiculosum mixed with
-glucosidase extracted from Almod) were investigated to increase the production of glucose from cellulose. Experimental result showed that optimum conditions fro pH, activity ratio of
-glucosidase to cellulase, concentration of mixed enzymes, concentration of cellulose as a substrate, and temperature range were 4.2, 0.4, 0.8, U/mL, 40 g/L, and 37
, respectively. In these conditions, quantities of glucose productions by using mixed enzymes were larger than those by using cellulase at optimum conditions.
Production Enhancement of Menthol in Suspension Cultures of Peppermint Using Cyclodextrin
KSBB Journal, volume 13, issue 1, 1998, Pages 26~30
The suspension cultures of Mentha piperita produce menthol which has very low solubility in water due to its hydrophobicity. This can be considered as a factor for its low production in the suspension suspension cultures. Cyclodextrin has the hydrophobic cavity inside the molecule in which menthol can be captured and allow to form a stable complex. The suspension culture of Mentha piperita showed 70% higher production enhancement in the medium containing 1.5%(w/v)
-cyclodextrin than the control.
-cyclodextrin had no adverse effect on the cell growth and showed the best result among
-cyclodextrins tested in terms of menthol production. We demonstrated that
-cyclodextrin can be used to enhance the production of menthol in the suspension cultures by capturing hydrophobic menthol into the cavity of cyclodextrin molecules.
Culture Condition of Entomopathogenic Nematodes Using Galleria mellonella Larva
KSBB Journal, volume 13, issue 1, 1998, Pages 31~37
A simple method for the in vivo production of third-stage infective juveniles(IJs) of Steinernema glaseri was developed. Using Galleria mellonella larvae, only IJs can be rapidly generated inadequate quantities for field application. The nematode inoculation concentration and incubation temperature were critically important. The most effective temperature for infectivity of Steinernema glaseri IJs to Galleria mellonella larvae was 33
. However, the total number of menatodes harvested at 25
about 66,000 IJs per larva was significantly greater than those at other temperatures. The optimal inoculation number of nematodes was 60 to 80 nematodes per host larva. The higher nematode inoculation concentration of 100 IJs per larva caused a rapid decrease in the total number of IJs harvested. As the inoculation medium pH increased, the number of IJs harvested increased and reached about 110,000 IJs per larva at pH 9.0. The pathogenicity of IJs decreased y increasing the salt concentration in the medium.
Simultaneous Saccharification and Pervaporative Fermentation of Cellulosic Biomass
KSBB Journal, volume 13, issue 1, 1998, Pages 38~43
Application of pervaporative extraction of ethanol to simultaneous saccharification and fermentation(SSF) of cellulose was investigated. From batch experiments, optimum cellulose substrate and enzyme loadings were found to be 10% and 15 IFPU/g cellulose, respectively. The cellulose conversion was lowered in fed-batch system due to the ethanol accumulation. The activity of the yeast Saccharomyces uvarum used in this study was significantly reduced at ethanol concentrations above around 40 g/L. From pervaporation experiments using PDMS membrane, ethanol was efficiently separated at 38
and 10 mmHg of a down stream pressure. The pervaporation unit with 240 cm
of surface area was combined into the SSF reactor. The continuous removal of ethanol by pervaporation during SSF resulted in an improved cellulose conversion. Within the scope of this experiment, ethanol yields in the pervaporative SSF and simple SSF were 68.3% and 56.6%, respectively. The permeate flux for SSF broth pervaporation was about one-half that for the pervaporation of aqueous ethanol solution. Accordingly, the development of a membrane with higher ethanol selectivity and flux will increase the feasibility of this technology.
Kinetics on the Specificity of Enzymatic Hydrolysis of Chitin
KSBB Journal, volume 13, issue 1, 1998, Pages 44~51
Hydrolysis and adsorption reversibility experiments were run for initial enzyme activity of 4.48, 9.65, 11.19 and 17.14U/mL at a temperature 30
. The chitin particle size corresponded to a mean particle diameter of 0.127mm, and the initial concentration of chitin was 10mg/mL. After approximately 2hrs, the enzyme activity remained constant in a speudo-steady state. The amounts in the bulk [E] and the amounts of enzyme adsorbed on the chitin surface [E] are plotted on Lineweaver-Burk plot to yield a linear relationship with a correlation coefficient of 0.99, a slope of 2.79cm
and an intercept of 0.08
/U. From this parameters, the values of [E
were calculated to be 12.5U/cm
and 34.88U/mL. respectively, Adsorption isotherm of the enzyme on the particles showed a well developed plateau of 1.35
. To determine the specificity of chitinase for crystalline chitin, the free energy of adsorption was measured, and its was determined as about -14.62~-18.8kJ/mol.
Electrochemical Determination of Immobilization Technique for Glucose Sensor Fabrication
KSBB Journal, volume 13, issue 1, 1998, Pages 52~57
The present work proposes a simple electrochemical method applicable to any immobilization processes of oxidase using a Clark type oxygen electrode as a base transducer. The present work suggests an optimal immobilization technique among three different methods of glucose oxidase(GOD) onto one side of
mthick blend membranes, composed o 80% of cellulose triacetate and 20% of polycaprolactone, on the basis of the maximum Michaelis-Menten parameter(Vm) determined by either steady state or transient analyses. The electrode system was made of disk type gold cathode(4mm diameter) and Ag/AgCl anode. One side of the blend membrane was in contact with the cathode surface while the other side was immobilized with GOD either in covalent-bond or cross-linked forms, the latter being covered by
m thick dialysis membrane of cellulose acetate. The resultant current density was on-line monitored by a potentiostat while glucose level was varied from 1 to 20 mM. The present study shows that direct cross-linking of GOD with glutaraldehyde was mostly preferred for fabrication of glucose sensor, on the basis of resultant kinetic parameters from either steady state or transient analyses.
Optimization of Culture Conditions for Extracellular Lipid Production from Rhodotorula glutinis K-501
KSBB Journal, volume 13, issue 1, 1998, Pages 58~64
An extracellular lipid-producing strain, Rhodotorula glutinis K-501, was isolated from soil. The extracellular lipid produced by the cell was found to have good and stable emulsifying activity. Factors affecting the extracellular lipid production were studied to optimize culture conditions for maximum production. Sucrose and ammonium sulfate were selected as best carbon and nitrogen sources, respectively because they gave the highest concentration of product. The optimum C/N ratio was 50. Optimum concentrations of
were 3.5, 1.0, 0.75, and 0.1g/L, respectively. The optimum temperature and initial pH were determined as
and 7.0, respectively. When the batch culture was conducted with a sucrose concentration of 60g/L under the optimized conditions, extracellular lipid was produced to a concentration of 8.1g/L with a high production yield of 51.9% based on dry cell weight.
Physiochemical Properties of Extracellular Lipid Produced by Rhodotorula glutinis K-501 as a Biosurfactant
KSBB Journal, volume 13, issue 1, 1998, Pages 65~70
The physiochemical properties of extracellular lipid produced by an oleaginous yeast, Rhodotorula glutinis K-501 were examined. From the analytical experiments, it was suggested that the extracellular lipid produced is glycolipidic compound. Critical micelle concentration and minimum surface tension of the extracellular lipid in aqueous solution were 89mg/L and 31dyne/cm, respectively. Surface tension was also constant throughout wide range of pH. The emulsifying abilities and dispersing power of the extracellular lipid were much greater than those of commercial surfactants such as Tween 80 and Triton X-100 by factors of 2-3 and 1.3, respectively.
Tissue Biosensor for Determination of
Channel Blocker in Chinese Drug and Seaweed (Porphyra yezoensis Ueda)
KSBB Journal, volume 13, issue 1, 1998, Pages 71~76
Tissue biosensor for mearsuring sodium channel blockers, such tetrodotoxin(TTX), saxitoxin (STX) and paralytic shellfish poisoning(PSP) consisted of frog bladder membrane, and
electrode. The proposed biosensor was applied to determine Chinese drug and dry or wet Porphyra yezonesis
channel blockers below the detection limit of the standard mouse bio-assay while the observed detection limit didn't cause human poisoning. The proposed biosensor system may be used for future
channel blockers monitoring within the marine environment.
Separation of Glutathione by Ion Exchange Chromatography
KSBB Journal, volume 13, issue 1, 1998, Pages 77~82
-glutamyl-L-cysteinylglycine, GSH) produced by microbial enzymes was separated by a liquid chromatography. In order to select a resin which would bind GSH efficiently, a batch adsorption experiment was carried out with GSH solution and various resins at pH 8.0 GSH bound to Q-sepharose and QAE-sephadex among anion exchange resins, but the latter was found not to be suitable because of the reduction of resin volume at high salt concentration. Preliminary experiments using a standard solution were carried out to separate GSH. GSH and
-glutamylcysteine were separated from the other constituents by applying step gradient of salt(NaCl) concentration. GSH was successfully separated from
-glutamylcysteine by applying Tris buffer containing 35mM NaCl. Chromatographic separation behaviors for the enzymatic product was similar to that for the standard solution. Separation yields of GSH from the standard solution and enzymatic product solution were 72.6% and 84.4%, respectively.
Bead-to-Bead Cell Transfer by Induction of Detachment of Anchorage Dependent HeLa Cells Grown on Macroporous Microcarriers
KSBB Journal, volume 13, issue 1, 1998, Pages 83~89
Using a cellulose macroporous microcarrier, HeLa cells were cultivated in 100mL spinner flask(Bellco Co., USA) and confluent cell laden microcarriers were subcultured by bead-to-bead cell transfer method. In macroporous mcirocarrier-HeLa system viable suspended cells played an important role in bead-to-bead cell transfer and that could be increased by use of RPMI-1640, a calcium-ion-reduced-media and high speed agitation. Successful bead-to-bead cell transfers were performed continuously three time in spinner flask. We applied this technique to produce recombinant Vaccinia virus which express
-galactosidase. Recombinant protein yield of bead-to-bead transferred culture was comparable to conventional microcarrier cultures that were inoculated by cells detached from T-flask. Although trypsinization is a useful method for subculturing microcarriers in some cases, that process adds quality control problem and handling steps to large scale cell production. There fore, bead-to-bead cell transfer technique offers another convenient and efficient scale-up method for continuous microcarrier cultures.
Process Development of Concentration of n-3 PUFAs from Fish Oil by Means of Lipase
KSBB Journal, volume 13, issue 1, 1998, Pages 90~95
Experiments on the process development for the concentration of polyunsaturated fatty acid (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil by using acyl chain specificity of Candida cylindracea lipase were performed. Five kinds of oils were hydrolyzed with Candida cylindracea lipase. Among then, Candida cylindracea lipase just had low activity on the PUFAs-rich fish oil. After the hydrolysis of fish oil, free fatty acid was removed and fatty acid components of glyceride mixtures were analyzed. When the hydrolysis was about 70%, the DHA content in the glyceride mixture was about three times more than that in the original fish oil. The EPA and stearidonic acid contents in the glyceride mixtures, however, were similar to that of the original fish oil. In this work, these results showed that the concentration process of PUFAs by using the acyl chain specificity of Candida cylindreacea lipase was effective in producing glycerides that contained a high concentration of PUFAs in good yield.
Molecular weight Control of Polyhydroxybutyrate (PHB) in Recombinant Escherichia coli
KSBB Journal, volume 13, issue 1, 1998, Pages 96~100
Two promoters (trc and P
) were inserted in PHA operon derived from Alcaligenes eutrophus to obtain high chain molecules of polyhydroxybutyrate (PHB) in recombinant Escherichia coli. Newly designed PHA operon was used to control the gene expressions of hydroxybutyric CoA and PHA polymerization, separately. Plasmids containing new synthetic operon was transformed into E. coli DH5
and analyzed for PHB production. Without induction of the PHA biosynthetic operon, PHA synthase which has low activity might supply low concentration of initiator during the polymerization reaction, resulting very high molecular weight of polymer. An increase of PHB average molecular weight was observed with decreased IPTG (isopropyl
-Dithiogalactosidase) concentration. When no IPTG was added to the culture of E. coli DH5
SJS1 which contained two promoters in PHA operon, high chain polymer having an average molecular weight of
was achieved. Analysis of the enzyme activities of PHA biosynthetic enzymes suggests that PHA synthase, the enzyme responsible for polymerizing 3-hydroxybutyric CoA, controls the molecular weight of PHB produced in vivo.
Biological Fixation of Carbon Dioxide by Synechocystis PCC 6803
KSBB Journal, volume 13, issue 1, 1998, Pages 101~107
Carbon dioxide is estimated to be responsible for 60% of the global warming effect, and this percentage is tending upward. Studies on removal and fixation of
in the flue gas are recognized as one of the important roles of the future biotechnology. Photobiological systems have considerably higher photosynthetic efficiency than conventional biomass system. The experiment for the photosynthetic fixation of
and the biomass production was performed with various initial cell concentration in a tubular photobioreactor and a bubble column
contactor with a gas sparger of
-enriched air(0.03~20%). Synechocystis PCC 6803 could grow at 10~20%
content under pH control. The highest specific growth rate, 0.0258
, was obtained at 5%
-air mixture. The maximum cell production rate, 0.2784 g/L.day, was obtained when the initial cell concentration was 0.45 g/L at 5%
-air mixture. The maximum cell concentration was 2.03 g/L in the tubular photobioreactor when the light intensity was
. s. This system showed 0.482 g
/L . day of the
Medium Fortification based on the Analysis of Amino Acids and Wastes in Hybridoma Culture
KSBB Journal, volume 13, issue 1, 1998, Pages 108~113
The cell growth and amino acid metabolism of a hybridoma cell line in T-flasks, spinner flasks, and a 2L bioreactor were compared. Similar growth and metabolic behaviour were observed for spinner flask and bioreactor cultivations, while those in T-flasks differed significantly. Through a detailed analysis of nutrients and wastes, 7 amino acids were found to be consumed to a much higher extent than the rest of the amino acids. Supplementing the based medium with selected amino acids, glucose, and vitamines increased the cell density by 70%. The addition of vitamines was found to increase the metabolic rates of glucose and lactate.
Effect of Solubilization Conditions on Molecular Weight Distribution of Enzymatically-Hydrolyzed Silk Peptides
KSBB Journal, volume 13, issue 1, 1998, Pages 114~118
The effects of fibron solubilization conditions on molecular weight distribution of enzymatically-hydrolyzed silk peptides were investigated. The weight-averaged molecular weights of silk proteins prepared by solubilization with calcium chloride, ethylenediamine and sulfuric acid were 41600, 3308, and 1268 dalton, respectively. Silk peptides in the average molecular weight range of 600-1200 dalton were obtained by protease treatment from solubilized silk fibroin. After the acid hydrolysis of silk protein using hydrochloric acid for 24 hr, silk protein was hydrolyzed to peptides whose average molecular weight and free amino acid content were 145 dalton and 80%, respectively. It was possible to control molecular weight distribution of silk peptides by the combination of solubilization and hydrolysis methods. Among the various treatment methods, acid solubilization followed by protease treatment had an advantage of molecular weight control for the peptide production.