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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 10, Issue 5 - Nov 1995
Volume 10, Issue 4 - Oct 1995
Volume 10, Issue 3 - Jul 1995
Volume 10, Issue 2 - May 1995
Volume 10, Issue 1 - Mar 1995
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Recovery and Utilization of Proteins and Lipids from Washing Wastewater in Marine Manufacture by Isoelectric Point Precipitation Method 1. The Coagulation Treatment for Washing Wastewatfr of Minced Mackerel Meat
KSBB Journal, volume 10, issue 1, 1995, Pages 1~8
A lot of water soluble proteins and lipids are released from minced mackerel meat and lost into the washing waste during the leaching process of Kamaboko or surimi manufacture. The removed proteins and lipids are not only an edible things but also a big burden for treating the wastewater. In order to recover the proteins from the effluent and to use as food stuff, the "pH-shifting" treatment, a modified isoelectric point precipitation method, was tried. This method is based on a myogen-aggregation phenomenon, which occurs when a solution of sarcoplasmic proteins is acidified or alkalified beyond the critical pH zone of 2∼3 or 12∼13 respectively and then neutralized. The maximum amount of precipitation was obtained by shifting the pH of the wastewater from original pH to isoelectric point (pH 4) or alkali pH 12 and then changing to neutral pH. The precipitates were easily collected by filteration or centrifuging at 10,000rpm. The oils which were only floating in the washing wastewater are easily recovered by seperating with oil separator after pouring. The recovered proteins were slightly denaturated during this pH shifting precipitation process, while the composition of amino acids was good balance as a food.
Continuous Production of Fructooligosaccharides from Sucrose by a Dual Immobilized Enzyme System of Fructosyltransferase and Glucose Isomerase
KSBB Journal, volume 10, issue 1, 1995, Pages 9~14
Continuous production of fructooligosaccharides from sucrose by a dual immobilized enzyme system of fructosyltransferase and glucose isomerase was studied in a column reactor. The optimal temperature and pH of the immobilized fructosyltransferase were
and 5.5, respectively. The activity of glucose isomerase was favorable as temperature and pH were increased within the ranges examined. However, both the immobilized enzymes were thermally unstable over
, suggesting that long-term operation of the dual immobilized enzyme column should be carried out below
. The optimum packing ratio of fructosyltransferase to glucose isomerase was found to be around 5/3. Under the optimized reaction conditions, the dual enzyme column was successfully operated for 40 days without any loss of initial enzyme activities, yielding 66% of fructooligosaccharides. Furthermore, the relative sweetness of fructooligosaccharides produced by a dual emzyme system was enhanced by 6% compared with that of fructosyltransferase alone.
In Vitro Propagation of Persimmon(Diospyros kaki) by Embryo Culture
KSBB Journal, volume 10, issue 1, 1995, Pages 15~22
The embryos(6-8mm) isolated from seeds of Diospyros kaki were cultured on Murashlge-Skoog(MS), Woody Plant Medium(WPM), Campbell Durzen(CD), Lictvay's Medium(LM), Kao-Michaluk(KM), Nitsch, White, Heller, Wolter-Skoog(WS) media. The results showed that MS and WPM media were most suitable to the development of embryos into plantlets with length of
and 5-6 leaves. However, when LM and KM media were used, the addition of 1 to
was required for the germination of the embryos. Superoxide dismutase (SOD) activities, one of the changing factors in leaves according to physiological status displayed to be exceptionally significant in the leaves of plantlets germinated from seeds in potting sand soil contrary to those of cultured embryos specially around germination period.
Transformation of Rice Embryogenic Cells by Electroporation Mediated Plasmid Uptake into Protoplasts 1. Plant Regeneration from Electroporated Protoplasts of Rice
KSBB Journal, volume 10, issue 1, 1995, Pages 23~29
Calli were induced from leaf base region of germinated rice(Oryza sativa L. cv. Nakdong) with high frequency of up to 65% on LS medium supplemented with
, 4-D in the dark at
. Embryogenic calli of pale yellow, globular type were selected and used for the initiation of cell suspension cultures in AA2 liquid medium with
GA3. Protoplasts were isolated from the embryogenic cell suspensions after 4 months of culture and then were electroporated with 400V/cm for 1 msec. Electroporated protoplasts divided with plating efficiency of 1.1% on PCM liquid medium supplemented with
kinetin and 10mM proline. The protoplasts-derived microcalli were cultured on
membrane fitter placed onto LS2.5 solid medium containing fine suspension cells as a feeder cells, for 2 weeks in the dark at
. After an additional 2 weeks of culture under fluorescent light of
, yellow calli of 2mm diameter were transferred to regeneration medium. Shoots were produced from the green spot of protoplasts-derived calli and plants were regenerated form protoplast-derived green calli with frequencies of 11∼33%.
Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. KM
KSBB Journal, volume 10, issue 1, 1995, Pages 30~37
Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32
, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.
Expression of Heterologous Promoters in Aspersillus oryzae
Hahm, Young-Tae ; Kim, Hee-Chung ; Batt, Carl A. ;
KSBB Journal, volume 10, issue 1, 1995, Pages 38~45
The expression of Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpdA) and trpC promoters in A. oryzae were compared using E. coli lacZ gents fusions. The specific activities of the expressed E. coli
-galactosidase in A. oryzae transformants containing the A. nidulans gpdA promoter were around 2,000 units per ug of protein. The specific activities of transformants containing the A. nidulans trpC promoter were very low, ranging from 10.5 to 52.3 units per ug of protein. These results showed that the expression of the A. nidulans gpdA promoter in A. oryzae was approximately 70 times greater than the A. nidulans trpC promoter. In western blot analysis, immunoreactive bands of a imlilar molecular weight as the E. coli
-galactosidase were observed in A. oryzae carrying the gpdA-lacZ fusion and to a lesser intensity in those carrying the tvpC-lacZ fusion. Southern analysis showed that the higher expression of the gpdA-lacZ fusion as compared to the trpC-lacZ fusion was not due a greater number of integrated plasmids.
Optimal Nutritional Requirements of Carrot Hairy Roots
KSBB Journal, volume 10, issue 1, 1995, Pages 46~54
The physiological characteristics of carrot hairy roots and suspension cells were examined based on their nutritional requirements. Inorganic nutrient (phosphorous and ammonium) requirements of carrot hairy roots were similar to those of suspension cells. Optimal sucrose concentration for the growth of hairy roods (7%) was different from that of suspension cells (3%). Since suspension cells were move easily affected by the environmental condition, e.g., osmotic stress, than hairy roofs which made the suitable growth condition for cells, it can be understood that optimal sucrose concentration for the growth of hairy roots was higher than that for the growth of the suspension cells. To investigate the roles of sucrose on the growth of hairy roots, the effects of sucrose on the fresh weight and dry weight was analysed by the addition of mannitol as an osmolicum. Sucrose acts also as an energy source for hairy roots rather than as an osmotic regulator, since the increase of dry weight was higher than that of fresh weight at the given sucrose concentration
Improvement of Alcohol Productivity by Means of Repeated Batch Fermentation
KSBB Journal, volume 10, issue 1, 1995, Pages 55~62
The functional relationship between the initial cell concentration and the ethanol productivity was investigated in the repeated batch fermentation of Sacharomyces cerevisiae ATCC 24858. The repeated batch fermentations were performed in the range of 60 to
of initial sugar concentration and 17.5g/
of initial cell concentration. The time of one batch fermentation was 1 or 2 hours and the batch fermentation was repeated ten times in every repeated formentaction. The functional relationship showed that the productivity increased non-linearly according to the increase of initial cell concentration regardless of initial sugar concentration. When the initial concentration of sugar was
and that of biomass was
, the fermentation was completed within one hour and its ethanol productivity was
.hr, the latter including the times of cell separation, pouring the new substrate into a flask and sampling. When the initial sugar concentration was
and the initial cell concentration
, the fermentation was also finished within one hour and its productivity was
hr, The maximum ethanol productivity for eight different repealed fermentations in this work was
Separation of Soybean Protein by Free-flow Electrophoresis
KSBB Journal, volume 10, issue 1, 1995, Pages 63~70
The effect of operating conditions on separation of soybean proteins in a home-made free-flow electrophoresis apparatus was investigated. Measurement of the pH, conductivity, and UV-absorbance(280 nm) were carried out at each run and the purity of the sample was tested with SDS-PAGE analysis. The soybean extract pretreated with Tris and boric acid was mixed with the amino acids composed of glutamic acid, histidine, arginine, glycine(1 mM each) with glycyl-glycine(2mM) and KCl(1mM). When the cellulose acetate was used as a compartment between the electrode and the buffer solution in the cell, pH distribution in the separation cell varied from 3.0 at the anodic side to 8.0 at the cathodic side and had two inflection point. The applied voltage was from 300V to 1000V and the separation was better at a higher voltage but the voltage was limited by the capability of the cooling system due to Joule heat. The proteins focused near the middle of the channel. From the change of pH and conductivity it was found that the ions in the channel moved out to the electrodes through the membrane. In the case when the concentration of the buffer solution was increased 5 times, proteins were focused at 300V. We could not increase up to the ten times of the concentration since the temperature difference between inlet and outlet was more than
and denaturation of proteins was expected. When ion-exchange membranes were used U-type pH distribution was set up due to the ionic polarization near the membrane. The commercial ampholytes, instead of the mixed amino acids showed not much improvements in purity of the separated sample.
Properties of Microbial Surfactant S-acid
KSBB Journal, volume 10, issue 1, 1995, Pages 71~77
Characteristics of S-acid and its derivatives, as biosurfactants, were investigated. Sodium salt of S-acid(S-lNa) effectively decreased the surface tension to near 30 dyne/cm and showed superior dispersing ability than commercial surfactants such as SDS and Tween 80. The emulsion made by S-lNa could be maintained even with increased concentration of calcium ion. Also the penetratlng ability of S-lNa was observed as effective as that of LAS. All derivatives of S-acid were degraded by microorganisms much faster than conventional surfactants. Among the derivatives, S-3Na prevented rust formation as effectively as commercial anti-rust agents did.
Citric Acid Production Using Encapsulated Aspergillus niger
KSBB Journal, volume 10, issue 1, 1995, Pages 78~88
The encapsulatpd A. niger grew up inside the capsule and mycelia penetrated through the pore of the capsule membrane. The mycelia on the capsule wall became loose when the carbon source and oxygen were deficient in the medium. On the contrary, the production rate increased and mycelia made a lump tightly when the carbon source and oxygen were sufficient. Namely, number of proper capsule of unit volume in the medium was existed. The phenomenon which was swelled of capsule membrane in cultivation could prevented by adding CaCl2 into the medium. According to the time adding CaCl2 into the medium, the production rate of citric acid was influenced. In case of adding CaCl2 into the medium at 7th day cultivation, the production yield of citric acid was increased about 40 percent higher than that of adding CaCl2 initially. The production yield of citric acid using encapsulated A. niger of flask culture was influenced with oxygen supply. The production yield of citric acid (
s) of the flask culture was increased 3.88 time by using T-flask instead of parafilm sealed flask. Therefore, the productivity and consumption rate concerning production which was taken carbon source were increased when oxygen supply was sufficient. The production of citric acid using encapsulated A. niger was increased average 30 percent higher than that of bead in between 6th and 13th day cultivation.
Molecular Approaches for Cloning of Important Higher Plant Genes
KSBB Journal, volume 10, issue 1, 1995, Pages 89~96
An Avabidofsis thaliana gene encoding phosphoribosyl anthranilate transferase is shown to be the gene that is defective in blue fluorescent trp 1 mutant plants. This gene, named PAT1, coding region is homologous to those for the phosphoribosyl anthranilate transferase from many microorganisms. This is due to a defect in tryptophan biosynthesis that leads to an accumulation of anthranilate, a fluorescent intermediate in the tryptophan pathway. PAT1 is a single-copy gene that complements all of the visible phenotypes of the different trp1 mutants. Experiments to determine the regulation of the PAT1 gene are in progress. The wild-type PAT1 promoter and several promoter deletions of PAT1 gene have been transformed into Arabidopsis tryptophan mutants. These constructs might identify promoter elements that control this patterns. We have isolated the homozygous lines in T3 seeds and analysed the protein levels using PAT antibody and PAT protein level increased two fold in pHSl07. Finally, the potential of using PAT1 as a selectable marker or visible reporter of gene expression is being explored.
Process Simulation for the Production of Porcine Growth Hormone Using CAD Program
KSBB Journal, volume 10, issue 1, 1995, Pages 97~104
A computer simulation of biochemical process was carried out using Macintosh-based BioDesignerTM developed at Bioprocess and Engineering Center(BPEC) of MIT. Based on the assumptions and flask culture experiments, a porcine growth hormone (PGH, Porcine Somatotropin) production process was simulated by a two-stage continuous culture. The economical and sensitivity analyses were evaluated for the scale-up production of PGH. A high return on investment (ROI, 104%/year) suggested that the process be profitable. However, sensitivity analysis indicated that ROI was dependent on the yield of PGH, selling price, dose and the achievement of projected market penetration.
Development of Biomolecular Device Using Biomolecular Film Part 1: Optical Biosensor to Detect the Ethanol Using Langmuir-Blodgett Film of Eilzyme Molecules
KSBB Journal, volume 10, issue 1, 1995, Pages 105~112
The fiber-optic biosensor using enzyme-immobilized Langmuir-Blodgett film is developed fort the measurement of ethanol. The enzyme, alcohol dehydrogenase, is immobilized at the molecular level on the arachidic acid monolayer using Langmuir-Blodgett film technique. Based on the ordered multisubstrate mechanism, the immobilized enzyme kinetics is investigated. The optical sensing system is proposed, and sensor signal is proportional to ethanol concentration and is related wish the number of enzyme layers. As the number of deposited LB film layer increases up to 20 1ayers, the high ethanol concentration of 45mM can be measured without the saturation of signal. Surface pressure-area isotherm is measured for the three-different charged-lipids. Arachidic acid is the most suitable for the adsorption of alcohol dehydrogenase based on electrostatic force.