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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
Isolation and Characterization of a Phenol-Degrading Bacteria
KSBB Journal, volume 13, issue 2, 1998, Pages 119~124
Twelve bacterial strains capable of growing on phenol minimal medium were isolated from iron foundry activated sludge by enrichment culture, and amount them, one isolate which was the best in cell growth and phenol degradation was selected and identified as Acinetobacter junii POH. The optimal temperature, initial pH and phenol concentration in the above medium were 3
, 7.5 and 1000 ppm, respectively. Cell growth of Acinetobacter junii POH dramatically increased 20 hrs cultivation-time and reached a almost stationary phsae 40 hrs cultivation-time then phenol was degraded about 98%. Cell growth was inhibited y phenol at concentrations over 1500 ppm. The isolate was resistant to several antibiotics as well as various heavy metal ions. The growth-limiting log P value of Acinetobacter junii POH on organic solvents was 2.9 in the LB medium. Therefore, it is suggested that Acinetobacter junii POH could be effectively used for the biological treatment of wastewater containing the presence of heavy metal ions and organic solvents.
Characteristics of Protein Chromatography by Affinity Membrane Mudule
KSBB Journal, volume 13, issue 2, 1998, Pages 125~132
Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70
determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20
for flow rates above 6 mL/min.
Comparative Analysis of Dissolution and Refolding Processes for Inclusion Body Protein Renaturation
KSBB Journal, volume 13, issue 2, 1998, Pages 133~140
and rhGH as the model proteins, the refolding performances of the published processes were evaluated and compared. Key engineering parameters such as the type of denaturant and this concentration, protein concentration in the refolding buffer, and pH and ionic strength of the buffer were experimentally investigated. Furthermore, the role of a co-solvent of surfactant type in aggregation reduction was also studied. Of the denaturants tested (8M urea, 6M guanidine HCI, 0.5% SDS), SDS at alkaline pH (9.5) and ambient temperature gave the highest recovery yield. The SDS process was effective in the refolding of observed where dissolution proceeded better under lower strength (10 mM) but aggregation was suppressed under higher strength (>50 mM.) When PEG-4000 and/or Tween were added as co-solvent or refolding-enhancing additive, 1.6-2 times higher yield was realized. The‘masking’of the hyrophobic patches located on the surface of the protein with the surfactant molecules was believed to be responsible for the considerable reduction in aggregation during refolding.
Removal of Methane Using a Three Phase Fluidized Bed Bioreactor
KSBB Journal, volume 13, issue 2, 1998, Pages 141~146
To remove the low concentration of methane biologically, a three phase fluidized bed bioreactor immobilized with Methylosinus trichosporium OB3b was used. Optimum pH, temperature and bed height for the operation were pH7.0, 30
and 150cm, respectively. For the inlet methane concentration of 100-400ppm and flow rate of 2-4L/min, the removal efficiencies of the bioreactor using the activated carbon as a carrier were the range of 54-71%, whereas those using the biosand were the range of 45-56%. It was found that activated carbon was more efficient than the biosand for the removal of methane. When aeration tank was equipped with the bioreactor, the removal efficiency increased to 6-13% and maximum removal rate obtained in the experiment was 1184mg.CH
Production of Chitosna Oligosaccharides Using Chitin-Immobilized Enzyme
KSBB Journal, volume 13, issue 2, 1998, Pages 147~154
Enzymatic hydrolysis using an immobilized enzyme was carried out to produce chitosan oligosaccharides (COSs) from chitosan effectively. Chitosanase was immobilized on eight different carriers by physical adsorption. The enzyme immobilized on chitin had higher activity than those immobilized on the other carriers in spite of its lower adsorption. The activity of chitin-immobilized enzyme was more than 90% of the original activity. Optimal temperature of the immobilized enzyme increased by about
and its thermostability was excellent in relatively wide range of temperature. But its effects of pH did not improve compared to the free enzyme. The immobilized enzyme produced 153 mg/g chitosan of the reducing sugar for 3hrs of hydrolytic incubation time. The total content of higher oligomers, tetramer to hexamer, among amount of total COSs obtained for 2hrs was more than 90%. In kinetic parameters for both enzymes, immobilized enzyme showed lower affinity for substrate and reaction rate than free enzyme, however, no reduction of the rate for high substrate concentrations. Consequently, chitin-immobilized could effectively hydrolyse chitosan and produce the higher COSs without activity decrease in comparison with the free enzyme.
Induction and Culture of Hairy Roots of Crotalaria sessiliflora L.
KSBB Journal, volume 13, issue 2, 1998, Pages 155~161
The hairy roots of Crotalaria sessiliflora were induced from the tissue segments infected with Agrobacterium rhizogenes ATCC 15834. The induced hairy roots were subjected to paper electrophoresis fro the detection of opine-positive clones which were considered to have been transformed. Mannopine and agropine were presented in hairy root clones while mannopine was presented in two hairy root clones. Eight hairy root clones were selected and cultured in MS, B5 and WP media. Each of hairy root clones was showed a difference in branch pattern and growth rate. The best culture medium and culture conditions of hairy roots were in
MS(3% sucrose, pH 5.7) liquid medium at 25
, 70 rpm under dark, the growth rate in
MS liquid medium was increased with 210-fold more than that of inoculated hairy roots and with 2-fold more than that in MS liquid medium. Also, the adequate condition for hairy root growth was such that concentration of KH
was 1.25mM and the ratio of NH
was 1 to 3 in MS medium. The presence of pyrrolizidine alkaloids, monocrotaline, in the hairy roots was detected by TLC.
Adsorption Characteristic of Endo I and Exo II Purified from Cellulase by Trichoderma viride on Celluloses with Different Crystallinity
KSBB Journal, volume 13, issue 2, 1998, Pages 162~167
The adsorption behaviors of two major cellulase components, endo I and exo II, from Trichoderma viride were investigated using
-celluloses with different correlation crystallinity index(Cc) as substrates. The adsorption of cellulase enzyme components was significantly affected by the reaction condition and the physicochemical properties of the cellulose. The
-cellulose was hydrolyzed in the presence of cellulase for various periods. The correlation crystallinity index of
-cellulose increased with increasing the hydrolysis time. The adsorption was apparently found to obey the first-order kinetics, and the adsorption activation energy(Ea) was calculated from the adsorption rate constant(ka). The value of adsorption rate constant for endo I was larger than that of exo II. This means that endo I are adsorbed more rapidly than exo II. With the increase in correlation crystallinity index, the values of the adsorption rate constants for endo I and exo II decreased, respectively. The activation energy for the adsorption of exo II on the cellulose also was larger than that of endo I. Also adsorption activation energy of endo I and exo II increased with an increase in the crystallinity of sample cellulose.
Combined Effect of Agrimonia pilosa Ledebour Extract and NaCl for Control of Escherichia coli O157:H7
KSBB Journal, volume 13, issue 2, 1998, Pages 168~173
Gamma irradiated and non-irradiated Agrimonia pilosa Ledebour were extracted by 70% ethanol. The combined effects of the Agrimonia pilosa Ledebour extract and NaCl on survival of Escherichia coli O157:H7 in tryptic soy broth were investigated. E. coli O157:H7 decreased ca 1 log cycle by the addition of 2% sample extract, and the anthbacterial activity was increased as the concentration of sample extract was increased. The irradiation effect of the sample on antibacterial activity was not observed. On the treatment of NaCl alone, E. coli O157:H7 was inactivated (ca 3~4 log cycle reduction within 48 hr) in more than 7% NaCl. The higher inactivation(ca 5 log cycle reduction within 48 hr) occurred in the presence of 2% sample extract and 5% NaCl than in the addition of each alone. The extracted antibacterial substance was stable in the pH range of 4.0 to 7.0, heat treatment at 121
for 15 min, and freezing at -18
and thawing at 37
. There fore, the sample extract, would substantially increase the food-safety in terms of E. coli O157:H7.
Studies on the Production of (10-Deacetyl) Baccatin III in Cell Cultures of Taxus baccata Pendula
KSBB Journal, volume 13, issue 2, 1998, Pages 174~180
Enhanced production of (10-deacetyl) baccatin III and related taxanes was observed in suspension cultures of Taxus baccata Pendula. six % of initial glucose and sucrose concentration increased 10-deacetyl baccatin III production 3.5 and 2.5 times, respectively. Methyl jasmonate, as an elicitor, increased taxane production. Time course changes of taxane production after methyl jasmonate addition showed that baccatin II and 10-deacetyl baccatin III were detected first and paclitaxel, 10-deacetyl taxol and cephalomanine were produced in sequence. Feeding experiments with
of benzoic acid increased 10-deacetyl baccatin III production 10 times. Baccatin III production was also increased 8 times by feeding of
of lysine as a precursor.
Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System
KSBB Journal, volume 13, issue 2, 1998, Pages 181~186
In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density
cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of
cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.
Mass Production and Accumulation Characteristics of Polyhydroxyalkanoates by Fed-batch culture of Alcaligenes eutrophus under Phosphate Limitation
KSBB Journal, volume 13, issue 2, 1998, Pages 187~194
For mass production of polyhydroxyalkanoates (PHA), high cell density cultures of Alcaligenes eutrophus by fed-batch culture under phosphate-limitation condition has been investigated. PHA accumulation by the regulation by the regulation of initial phosphate concentration could be automatically induced, and high density cell culture above 200 g/L also could be successfully produced. The production of Poly-
-hydroxybutyrate (PHB) and dry cell weight increased with increasing the initial phosphate concentration. When the initial concentrations of phosphate were in the ranges of 1.5~4.5 g-PO
/L, PHB and dry cell weight obtained were 83~266 g/L and 61~216 g/L, respectively, and PHB productivity was in the ranges of 1.35~3.10 g/L.h. When a mixture of glucose and propionic acid is used as carbon sources, poly(3-hydroxybutyrate-co-poly-3-hydroxyvalerate), P(3HB-co-3HV), could be also successfully produced under phosphate limitation condition. When the mole ratio of propionic acid to glucose in the feeding solution is 0.22, a final dry cell weight of 150 g/L and a P(3-HB-co-3HV) of 90 g/L were produced. Morphological changes and size distribution of PHB granules synthesized in A. eutrophus under phosphate-limitation condition are determined by TEM during the course of fed-batch. Mean granule diameters of PHB produced are in the range of 0.36~0.39
m, and mean cell size was elongated from 0.54~0.59
m to 0.83~0.89
m. Phosphate concentration in media did not affect size distribution of PHB granule and cell.
Pruification and Characterization of Cholesterol Oxidase Produced by Rhodococcus sp. 3T6-5Mj isolated from Changran-jeot
KSBB Journal, volume 13, issue 2, 1998, Pages 195~202
The cholesterol oxidase was purified from the culture broth of Rhodococcus sp. 3T6-5Mj strain by procedures involving filtration, acetone precipitation, DEAE-Sephadex A-50, and cholesterol affinity column chromatography with a recovery of 15% to specific activity of 25.6 units/mg. The molecular weight of the enzyme was estimated to be 52,000 daltons by SDS-PAGE. Optimum pH and temperature for the enzyme activity were approximately pH 7.0 and
respectively. The Michaelis constant (Km) for cholesterol was found to be
M. The enzyme showed a high substrate specificity for
-hydroxysterols and the relative oxidation rates were 100% for cholesterol, 89% for campesterol, 55% for stigmasterol, etc. Amino acid analysis showed that the enzyme protein was composed of 440 amino acid residues without cystein and tryptophan.
Studies of Cyclosporin A Biosynthesis under the Conditions of Limited Dissolved Oxygen or Carbon Source in Fed-batch Culture
KSBB Journal, volume 13, issue 2, 1998, Pages 203~208
We investigated the effects of dissolved oxygen (D.O.) and fructose (C-source) on cell growth and biosynthesis of cyclosporin A (CyA) produced as a secondary metabolite by a wild-type filamentous fungus, Tolypocladium inflatum. This was performed by controlling the level of D.O. and the residual C-source, as required, through adjustment of medium flow rate, medium concentration and agitation rate in fed-batch cultures. CyA production was furned out to be maximal, when D.O. level was controlled around 10% saturated D.O. and concentration of the C-source was maintained sufficiently low (below 2 g/L) not to cause carbon catabolite repression. Under this culture condition, we obtained the highest values of CyA concentration (507.14 mg/L), Qp (2.11 mg CyA/L/hr),
(0.49 g DCW/g fructose),
<(22.56 mg CyA/g fructose), and YTEX>$_p/x$ (48.31 mg CyA/g DCW), but relatively lower values of cell concentration (11.98 g DCW/L) and cell productivity (0.043 g DCW/L/hr), in comparison with other parallel fed-batch fermentation conditions. These results implied that, in the carbon-limited culture with 10% saturated D.O. level, the producer microorganism utilized the C-source more efficiently for secondary metabolism.
De-emulsification of Petroleum Emulsion Using Nocardia amarae
KSBB Journal, volume 13, issue 2, 1998, Pages 209~213
The characteristics of de-emulsification of pertroleum emulsion by Nocardia amarae were investigated. Insoluble medium containing n-hexadecane was more effective than soluble medium in de-emulsification of emulsion containing diesel and bunker C as the organic phase. Emulsion made by the addition of xanthan or bioemulsifier was de-emulsified by N. amarae, and longer culture age was effective. In low viscosity range, organic phase with longer carbon chain was more effective. The contact, angle between bacterial film and water droplet in air increased from 16 degree for 4 day culture age to 26 degree for 15 day. The contact angle between bacterial film and water droplet in kerosene, n-heyxane or n-hexadecane also increased to greater than 100 degree after 3 day culture age. The hydrophobicity of bactgerial film increased according to the culture age.
Overproduction of Sodium Gluconate Using the Recombinant Aspergillus niger
KSBB Journal, volume 13, issue 2, 1998, Pages 214~219
Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400
of hyg.B while the minimal media inhibited mycelial growth completely at 200
of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.
Degradation of Phenolic Resin, Resole by Microbial Consortia
KSBB Journal, volume 13, issue 2, 1998, Pages 220~222
Three microbial consortia were screened for their ability to degrade phenolic resin, resole as a sole carbon source. These microbial consortia were derived from soil samples collected from a phenolic resin manufacturing plant site. Among the consortia, the test consortium, designated as MS2, displayed approximately 70% degradation of the substrate, 100 mg of resole per liter, within the fist twelve days of incubation but the degradation was inhibited. During the incubation period, pH was decreased from 7.0 to 2.7, and the resole degradation became inhibited under the conditions. UV-scans of spent culture showed that the wavelength of maximum absorption was 261 nm for resole.