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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 6, Issue 4 - Dec 1991
Volume 6, Issue 3 - Nov 1991
Volume 6, Issue 2 - Aug 1991
Volume 6, Issue 1 - Apr 1991
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High Frequency Electroporation-transformation of Coryneform Bacteria Grown in the Medium with Penicillin-G
KSBB Journal, volume 6, issue 3, 1991, Pages 223~230
Using the shuttle vector pECCGl between Escherichia coli and Corynebacterium glutamicum and C. glutamicum strain JS231 grown in the medium supplemented with penicillin-G, which inhibits the formation of cross-links in the peptidoglycan of bacterial cell wall, various parameters involved in electroporation system including resistance, electric field strength, capacitance, DNA concentration, and cell density were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Using cells grown with 0.3U/ml of penicillin-G and harvested at A600 of 0.7-0.8, transformation efficiencies of 107-l08 transformants/
of DNA with Corynebcctertum glutamicum strain JS231 and wild type ATCC13032 were achieved under conditions of 12.5kV/cm of electric field strength, 400 ohms of resistance,
F of capacitance, 3
108 cells per transformation(1.2
1010 cells/ml) and 100ng of plasmid DNA per transformation.
Characteristics of a Bioreactor Using Perfluorocarbons and a New Impeller
KSBB Journal, volume 6, issue 3, 1991, Pages 231~240
Recently, developments of large scale and high density cell culture methods have been the objects of many researches, because the demand of various pharmaceutical products produced by animal cell culture has been rapidly increasing. The cell culture equipment should have the requirements such as sufficient oxygen transfer and mixing, low shear stress and surface tension, and small foaming. In order to develop a proper bioreactor meeting these requirements simultaneously, a perfluorocarbon having high solubility of oxygen was sprayed into the medium as an oxygen carrier instead of air. Also, a new impeller was developed and combined together with the perfluorocarbon spraying system so as to design a new bioreartor for cell cultivation. The new impeller had better characteristics of mixing and oxygen transfer than the paddle and cell-lift impellers based on the same, shear rate. But, it was observed that the volumetric oxygen transfer coefficient of the new bioreactor decreased with increasing cell density during E. coli fermentation.
Active Transport of Anions through Synthesized Polymer Membrane with Pyridine as Fixed Carrier
KSBB Journal, volume 6, issue 3, 1991, Pages 241~247
The Poly (4-vinyipyridine-co-styrene) membrane containing Pyridine as fixed carrier was synthesized and characterized. And the active transport mechanism of Cl- and
with changing concentration of
was transported not only by a symport mechanism with
transfer but also by an antiport mechanism with
was transported only by a symport mechanism with
transfer. Observing the initial flux of anions, salt formation constant between ions and membrane (K), and diffusion coefficient in membrane (D) were calculated as follows: for
Improvement of Process for Sorbitol Production with Cetyltrimethylammoniumbromide Permeabilized Cells of Zymomonas mobilis through Glutaraldehyde Crosslinking
KSBB Journal, volume 6, issue 3, 1991, Pages 249~254
Permeabilization of Zymomonas mobilis with CTAB(Cetyltrimethylammoniumbromide) was investigated in order to obtain stable process for sorbitol production in the immobilized system. The optimum conditions for sorbitol formation were obtained in the case of using cells treated with 0.2% CTAB at
for 10 min. Permeabilized cells were treated with glutaraldehyde to cross-link the internal enzyme for the improvement of the enzyme stability. In this way, no significant loss of enzyme activity was apparent during 30-day operation in a continuous process. The productivity of the continuous process at dilution rate 0.2h-1 was 6.51g/1/h for sorbitol. The CTAB permeabilized cells could be used to produce sorbitol in the long term continuous process.
The Optimization of the Composition of Nitrogen Source in the Medium of Alcohol Fermentation of S. cerevisiae
KSBB Journal, volume 6, issue 3, 1991, Pages 255~261
The effect of concentration of yeast extract and NH4Cl in the mediun of alcohol fermentation of S. cerevisiae ATCC 24858 on the fermentation characteristics, specific growth rate, sugar conversion, alcohol productivity was experimentally investigated. Regardless of initial sugar concentrations, the values of the above three characteristics increased with augument of concentration of yeast extract. However, the increasing tendency ceased above a certain concentration. The concentration of NH4Cl had little effect on the change of the three characteristics. The functional relationships between the concentration of yeast extract and the characteristics were different according to the initial sugar concentrations, but those between the ratio of yeast extract concentration to initial sugar concentration and the characteristics could be expressed as same forms respectively regardless of initial sugar concentrations. Also the values of the three characteristics approached to the maximum values around 0.085 of the ratio, but did not increase any more above 0.1 of the ratio. We have come to conclusion that the optimum ratio of the yeast extract concentration to the initial sugar concentration was about 0.085 and the ratio should not be decided as greater than 0.1 in the medium of alcohol fermentation of S. cerevisiae ATCC 24858.
Adaptive Control of Cell Recycled Continuous Bioreactor for Ethanol Production
KSBB Journal, volume 6, issue 3, 1991, Pages 263~270
The optimal cell concentration and dilution rate for maximum ethanol productivity were obtained using dynamic simulation in cell recycled continuous bioreactor. The good control performance was observed using rule-based STR (self-tuning regulator) compared to conventional STR. Rule-base contained the scheme to implement the STR in an efficient on-off way and the scheme for the controlled variable to reach the optimal value in a short time. Since a mathematical model was used to analyze and estimate the changes of the state variables and the parameters, it was possible to understand the physical meaning of the system.
Biosynthesis of Unnatural Phospholipids by Phospholipase D: I. Synthesis in A Emulsion System
KSBB Journal, volume 6, issue 3, 1991, Pages 271~279
Phosphatidylglycerol(PG) and two unnatural phospholipids, phosphatidylethyleneglycol (PEG) and phosphatidylpropyleneglycol(PPG), were synthesized from ovolecithin using cabbage phospholipase D(PLD) in a emulsion system. Optimum pH and temperature for the enzymatic synthesis of PG, PEG and PPG in the emulsion system was 5.0-5.6 and 37
, respectively. The maximum activity for transphosphatidylation was obtained with 30-80 mM Ca++. Addition of 25% glycerol was required to convert completely ovolecithin to PG, whereas 16% glycerol was sufficient to attain the highest rate of conversion for both PEG and PPG syntheses, the highest conversion rate was obtained with addition of either 10% ethyleneglycol or propyleneglycol. However, the concentration of alcoholic acceptor should be increased up to 20% to improve selectivity up to 100% for PEG or PPG synthesis. Identification of PEG and PPG was made by analyzing the polyvalent alcohols released after their hydrolysis by HCl or PLD.
Biosynthesis of Unnatural Phospholipids by Phospholipase D: II. Effect of Organic Solvents on Transphosphatidylation
KSBB Journal, volume 6, issue 3, 1991, Pages 281~288
This research was carried to investigate the effects of several organic solvents on the enzymatic transphosphatidylation in emulsion and two-phase solvent systems. The solvents having a similar dielectric constant with diethylether were effective for the enzyme activity. Diethylether and butylacetate were the most effective solvents, when added 12-15%(v/v) and 10-40%(v/v), respectively, for the synthesis of phosphatidylglycerol, phosphatidylethyleneglycol and phosphatidylpropyleneglycol. In the emulsion system, the size of ovolecithin liposome was increased and the clearness of the phospholipid bilayer was reduced as increasing the diethylether concentration. In the twophase solvent system, the rapidest reaction was obtained when water-organic solvent ratio was close to 1. The ratio of aqueous phase. however, should be lowered to 37% to gain the sole product of transphosphatidy1ation, without phosphatidohydrolysis.
Transfer of Insecticidal Toxin Gene in Plants: 2. Subcloning of B. thuringiensis Insecticidal Protein Gene and Rapid Plantlet Regeneration from Nicotiana tabacum Protoplast and Callus
KSBB Journal, volume 6, issue 3, 1991, Pages 289~297
The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7
105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.
Light Mediated Yeast Cell Growth and Metabolism
Dowd Jr, Christopher J. ; Tanner, Robert D. ;
KSBB Journal, volume 6, issue 3, 1991, Pages 299~307
In this paper the effect of light on non-aerated Baker's Yeast(Saccharomyces cereuisiae) production and the protein excretion to the extracellular fluid is studied. Previous results in our laboratory indicate that at pH=5 and T-32
yeast may be affected by light, but those differences seem to be within statistical variation of the data. In this paper, cell and extracellular protein concentrations along with redox potential are monitored for batch fermentations in the presence and absence of light at pH levels of 3 and 5 and at 31
, in order to explore whether possible light effects can be more readily discerned at lower pH values. Yeast particle size distributions are also determined over the course of fermentation using a particle counter in order to add one more measuring tool to our usual cell and total protein measurements. An apparently noticeable difference in the redox potential is observed between the light and the dark runs for early times for the pH=3 runs. The particle size distributions show differences in the particle diameters between light and dark runs at pH=3, but those differences fall within one standard deviation of the mean particle diameters.
Continuous Hydrolysis of Cod Skin Gelatin in an Ultrafiltration Reactor
Kim, Se-Kwon ; Byun, Hee-Guk ; Cheryan,Munir ;
KSBB Journal, volume 6, issue 3, 1991, Pages 309~319
A continuous stirred tank membrane reactor(CSTMR ) was developed and optimized for the production of cod skin gelatin hydrolyzates using endo-protease Alcalase. A experimental design methodology was used to optimize the four performance variables: enzyme concentration, substrate concentration, permeate flux and reactor volume. All four variables studied had an effect on substrate conversion, with enzyme and substrate concentrations being predominant. Conversion increased with the increase in enzyme concentration, with the decrease in substrate concentration, at high volumes and low flux. A strong interaction was observed between enzyme and substrate concentrations and smaller interactions between enzyme and flux and substrate and flux. The optimum operating conditions for the CSTMR process for an initial substrate concentration for 10% were
, pH 8, flux 7.3ml/min, residence time 82 min, and Alcalase to substrate ratio 0.02(w/w). A gradual decay in reactor activity during 8 hrs was 2.1% conversion/hr. Enzyme leakage through the 10, 000 MWCO membrane was 16% at
and 12% at
, 6hrs. However, there was no apparent correlation between enayme leakage and substrate conversion. The Km value for the CSTMR was 20 times higher than the batch reactor. The productivity(expressed as mg product/mg enzyme) of the CSTMR was more than six fold higher than the batch at
. The hydrolyzate was non-bitter.
Effects of Medium Components on Microbial Production of L-Phenyralsnine
KSBB Journal, volume 6, issue 3, 1991, Pages 321~325
In thisstudy, effects of medium components on microbial production of L-phenylalanine by Corynebacterium glutamicum were investigated. The effect of carbon source on the production of L-phenylalanine was significant. Molasses enhanced the production of L-phenylalanine compared to sucrose, glucose, fructose, or their mixture. It was noticed that trace salts were required for the cell growth and product formation in the minimal medium, but excess amounts of trace salts had no effect on the production of L-phenylalanine. It was also found that optimum amounts of biotin and thiamine were required for the cell growth and the production of L -phenylalanine.