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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 9, Issue 5 - Dec 1994
Volume 9, Issue 4 - Oct 1994
Volume 9, Issue 3 - Sep 1994
Volume 9, Issue 2 - Jun 1994
Volume 9, Issue 1 - Mar 1994
Selecting the target year
A Study on the Pullulan Production Using Whey
KSBB Journal, volume 9, issue 2, 1994, Pages 91~97
In this study, Pullulan Production from whey medium by Aureohsidium pullulans was investigated. When the Pullulan production from whey medium was carried outs final cell concentration was similar to sucrose basal medium but Pullulan concentration was less than 5g/
. For pullulan production from whey medium, adaptation culture technique was tried on lactose and galactose base medium When the adaptation culture technique was not applied, the maximum concentration of pullulan was 3.4g/
for lactose, 2.5g/
for galactose. Adapted strains produced 10~12g/
from lactose, 10~11g/
from galactose. The pullulan production from lactose basal medium was 13.5g/
for lactose adapted strains and 18.6g/
for galactose adapted strains. When the adapted strains were inoculated on whey medium, maximum pullulan production was obtained at initial pH 3.0.
A Study on the Chromatographic Separation of Proteins Using Fibrous Beds(I) -Adsorbent Fiber Manufactures and Data Handling-
KSBB Journal, volume 9, issue 2, 1994, Pages 98~103
A bed configuration wherein sheets of modified fibrous polyethylene are potted within a Millipore Filter Cartridge matrix has been developed. Polyethylene fibers form sturdy beds but the native hydrophobicity and inertness of polyethylene have precluded their use in protein chromatography The polyethylene fibers used in this system were modified by plasma oxidation and further derivatization. The resulting fibers are hydrophilic, bind protein reversibly and serve as an anion-exchange stationary phase. Separation of Bovine Serum Albumin on this bed, as well as results of basic studies on capacity and reversibility of binding within a fibrous bed and experimental data handling system are shown.
Continuous Ethanol Production by Tower Fermentor
KSBB Journal, volume 9, issue 2, 1994, Pages 104~107
A cone-type tower fermentor packed with Sacchromyces uvarum was employed to examine the continuous ethanol fermentation process. The maximum yeast concentration in the cone-type tower fermentor was 37.5-39.5g/
, the maximum ethanol productivity at the dilution rate of
. hr and the average ethanol yield was 0.48g EtOH/g glucose, which was 94% of the maximum theoretical yield. It was concluded that a cone-type tower fermentor might offer better perspectives for continuous ethanol fermentation.
Kinetic Analysis of Transglycosylation Reaction of Stevioside Using Raw Starch as a Glycosyl Donor
KSBB Journal, volume 9, issue 2, 1994, Pages 108~114
Kinetic equations for transglycosylation of stevioside in the attrition coupled reaction system using raw starch as a glycosyl donor were derived considering that the reaction was carried out through two steps; production of cyclodextrin(CD) from raw starch in the attrition coupled reaction system and then transglycosylation of glycosyl residues to stevioside from produced CD. Kinetic constants of derived equation were evaluated. The simulation result showed that the derived kinetic equations could predict the experimental data reasonably well and that can be utilized for optimization and scale-up of transglycosylation reactor and process developments.
Separation of Lentil Lectin Using Free-Flow Electrophoresis
KSBB Journal, volume 9, issue 2, 1994, Pages 115~121
A Purification device with 30-channel free-flow electrophoresis was assembled to treat samples of 240m1 volume for purification of lentil lectin (LcH) from lentil seeds with no impurities in a silverstained PAGIEF gel. HEPES(50mM)-Ttis(50mM), Cycloserine(50mM)-urea(3M), Histidine(50mM)-urea(3M) were used as ampholytea among which Histidine(50mM)-urea(3M) (pI 7.65) was found best in resolution. LcH is known to be present in the form of LcH-A, LcH-B and the complex of the two. The complex, however, disappeared when urea was added in the ampholytes, which suggested that the complete purification of two isolectins is possible using the present multistep purificaton device.
Isomalto-oligosaccharide Production from Maltose by Intact Cells of Aureobasidium pullulaans
KSBB Journal, volume 9, issue 2, 1994, Pages 122~126
A new method for the production of isomalto-oligosaccharides from maltose was investigated using intact cells of Aureobasidium pullulaans which had been known to produce fructo-oligosaccharides. The cells showed transglucosylation activity producing isomalto-oligosaccharides at high concentrations of maltose, while they showed a hydrolytic activity at low concentrations of substrate when cultivated at
. The optimum reaction conditions for the isomalto-oligosaccharide production were as follows: substrate concentration, 500g/l maltose; pH, 4.5; temperature,
; cell dosage, 10 unit per gram substrate. Under optimized conditions, the maximum yield of isomalto-oligosaccharides achieved was around 48% (w/w). At the early period of reaction, panose was selectively produced from maltose, and thereafter isomaltotriose was synthesized by utilizing panose as a substrate when maltose consumption was discontinued.
Production of Fructooligosaccharides by an Amyloglucosidase
KSBB Journal, volume 9, issue 2, 1994, Pages 127~132
A new method of fructooligosaccharides production was investigated by an amyloglucosidase using sucrose as a substrate. Optimum reaction conditions were as follows: sucrose concentration, 700g/
; pH 5.5; temperature,
; enzyme dosage, 48 units per gram sucrose. At the optimized reaction conditions, 41.5% of fructooligosaccharides were produced after 25 hours. A hydrolyzing activity was stronger than transfructosylting activity at low sucrose concentrations, resulting in low production rate of fructooligosaccharides. The optimum pH and temperature in both transfructosylating(pH 5.5,
) and hydrolyzing activity(pH
)were significantly different from each other. The amyloglucosidase also utilized fructooligosaccharides as a substrate and glucose seemed to be an inhibitor.
O1igosaccharide Formation and Production of Transfructosylase and Transglucosylase by Aureobasidium pullulans
KSBB Journal, volume 9, issue 2, 1994, Pages 133~139
Oligosaccharide formation and the production of transfructosylase and transglucosylase by Aureobasidium pullulans were studied in sucrose or maltose media, respectively. The initial uptake rates of substrate in sucrose-rich media were faster than that in maltose-rich media, also most parts of oligosaccharides formed and other monosaccharides released were utilized progressively as substrate during the cultivation periods. However, when the initial amount of sucrose was raised to
, high concentration of monosaccharides were liberated, consequently high-level fructose was accumulated unused during fermentation. The biggest molecule of oligosaccharide synthesized was hexasaccharide in all cultivation media examined, of which the organism could not utilize isomalto-oligosaccharide of DP6 synthesized in a maltose-rich medium. The maximum amount of oligosaccharides produced was
of sucrose and
of maltose were used as initial substrate. From the early stage of growth both fructooligosaccharides and isomalto-oligosaccharides were synthesized and progressively utilized as substrates during the fermentation. Based on the experimental results, it was suggested that maltose could induce both transfructosylase and transg1ucosylase, whereas sucrose was unable to slimulate transglucosylase formation.
Mannitol Production by Aureobasidium pullulans
KSBB Journal, volume 9, issue 2, 1994, Pages 140~146
Aureobasidium pullulans produced high concentration of polyols extracellularly in the media of sucrose, glucose and mannose as sole carbon source. Mannitol was the main polyol produced during the late exponential and stationary phases of growth together with small quantities of glycerol. Sucrose and glucose were rather rapidly metabolized to mannitol among carbon sources examined where the initial glucose concentration showed no difference in the amount of mannitol. In contrast 20%(w/v) of sucrose was the most appropriate concentration tested. However, the yield of mannitol based on substrate used(
) was independent on the initial concentration, and the mean value of mannitol yield in 10% glucose and sucrose media was 0.144 and 0.188, respectively. Mannitol production was reduced in response to an elevated water stress imposed by salts within the range from 0.25 to IM of NaCl or KCl as stress solutes. However, glycerol contents and its ratio to mannitol were increased at the conditions of high salinity. Based on the results, extracellular mannitol produced by A. pullulans probably resulted partly from osmoregulation(in case of glycerol) and mainly from, as known to occur in most of fungi, enzymatic reduction of the corresponding hexoses through phosphate pathway.
Production of Bluish Purple Pigment from Streptomyces californicus KS-89
KSBB Journal, volume 9, issue 2, 1994, Pages 147~156
A study was carried out for production of a pigment : bluish purple, using a mutant Streptomyces californicus KS-89-7. The mutant was induced from Streptomyces californicus KS-89 with N-methyl-N-nitro-N-nitrosoquanidin(MNNG). It was immobilized on an inert substance made of colloidal sillica and 3.5% sodium alginate with 1 to 10 ratio. The diameter of inert bead was 2mm, and number of immobilized mutant spore was approximately
/ml. It was packed in a column reactor and fermentation was conducted with a substrate made of soluble starch 1%, glycerol 1.0%, sodium glutamate 0.1%, sodium nitrate 0.05%, L-prolin 0.025% and with some trace elements. The aeration for production of the pigment was 2.5m1/min with semi-continuous fermentation. The pigment production reached at peak on 8 days of fermentation, and the mutant produced the pigment 1.8 times more than its parent strain with the maximum pigment production of
. The pigment production continued for 24 hours of fermentation, and at the end of the fermentation the mutant produced the pigment
High Yield Saponin Production by Mass Cultures of Ginseng Transformed Tissue I. Induction, Culture of Transformed Tissue and Selection of High-Saponin-Producing Clones in Ginseng
KSBB Journal, volume 9, issue 2, 1994, Pages 157~164
Hairy root clones of Panax ginseng were established by selection of some hairy roots formed on the leaf, stem and root segments transformed with Agrobacterium rhizogenes strain
. The transformed roots grew well in MS medium under the dark condition. To confirm the transformation with Ri-T-DNA, dot blot hybridization and opine analysis were Performed. Among four hairy roots induced from different part of ginseng, the HB3 hairy roots were examined for selection of high-saponin-producing clones. Four clones isolated from HB3 hairy root cultures displayed various phenotypes characterized by growth and total saponin content. Maximum growth was obtained for cultures of HB3-10 clone and the content of total saponin was 0.55 wt%. However, higher amount of total saponin was obtained with HB3-2 clone cultures(0.74 wt%) in spite of lower growth. Dot blot hybridization confirmed the introduction of Ri-T-DNA in the plant genome. In the opine test, agropine and mannopine were detected from all hairy root clones.
The Structural Characterization of Recombinant Bovine Somatotropin Expressed in Escherichia coli
KSBB Journal, volume 9, issue 2, 1994, Pages 165~173
In this paper we have described the structural characterization of recombinant bovine somatotropin produced in Escherichia coli. Recombinant bovine somatotropin consists of 191 amino acid residues with a calculated molecular weight of 21,802 Da. For fragmentation of recombinant bovine somatotropin, we have used trypsin, Staphylococcus aureus V8 pretease, CNBr, and mild acid hydrolysis method. Digestion and cleavage with these proteases and chemicals yielded peptides of various size for amino acid sequence determination. The N-terminal sequence analysis was carried out up to thirty residues. Because the design of the recombinant bovine somatotropin gene for expression was such that the coding sequence begins with an initiation codon, AUG, before Ala, the first amino acid of bovine somatotropin, we could expect the initial amino acid as N-formyl Met. But the first amino acid of this protein, expressed in E. coli cells as inclusion bodies, was Ala. And the amino acid composition of RP-HPLC purified recombinant bovine somatotropin was determined and no essencial difference was observed. The amino acid sequence of the recombinant bovine somatotropin was identical to that predicted from its recombinant gene. There was no processing or replacement of amino acid residues in recombinant bovine somatotropin expressed in E. coli. The hydropathy plot of recombinant bovine somatotropin revealed a hydrophobic region at the NH2-terminus and hydrophilic region at the COOH-terminus. The E. coli expression system is thought to be valuable for the expression of recombinant bovine somatotropin because protein was processed to remove the N-terminal Met residue by methionyl-aminopeptidase autonomously.
Isolation and Characterization of Chitin from Crab Shell
KSBB Journal, volume 9, issue 2, 1994, Pages 174~179
Chitin was isolated from crab shell wastes and characterized for its chemical and physical properties. White powdered chitin was obtained through demineralizaticn, deproteinization and decoloration process. The contents of inorganics was less than 0.5%, whereas protein and lipid were almost removed. The results of IR spectroscopic analysis for the isolated chitin showed similar characteristics with that of Sigma product. Degree of deacetylation of purified chitin was significantly higher than Sigma product and viscosity average molecular weights was
. SEM analysis showed that the obtained chitin had the fibril shaped morphology.
Effects of Gas Recycle on Plant Cell Growth and Secondary Metabolites Production in Airlift Fermentor
KSBB Journal, volume 9, issue 2, 1994, Pages 180~185
The productivity of alkaloid in the airlift fermentor operation was less than that of suspension coltures of Eschscholtzia californica cells in the shake flask. To overcome the productivity reduction, a gas recycle airlift fermentor was developed because the gas-stripping in normal airlift fermentor was believed to play a significant role for productivity reduction. The alkaloid content in the gas recycle system with Eschscholtzia californica suspension cells was 2.7 times higher than that of normal airlift fermentor. The productivity of alkaloids and
concentration were affected by the volume of gas reservoir in the gas recycle airlift fermentor.
Analysis of Taxol and Related Compounds in Ullung Island Yew (Taxus cuspidata var. latifolia)
KSBB Journal, volume 9, issue 2, 1994, Pages 186~190
The content of taxol and related compounds in various tissues of native yews( T. cuspidata var. latifolia) grown in 5 locations of Ullung Island were analyzed. A considerable range of vanation in the content was observed in the needle and bark collected from different trees located at the same area as well as at five different areas. Taxol content was much higher in the needle(0.017% on the dry weight basis) than those in the bark, whereas the content of 10-deacetyl baccatin III(10-DAB III) was slightly higher in the bark(0.073%), regardless of the location of the trees collected. Particuayly, the needle collected from the Hyunpo area, which located in North-west part of Ullung Island, contained the highest level of taxol(0.024%) exceeding the reported level in dried barks of Pacific yew and also somewhat higher level of its Precusor, 10-DAB III(0.049%). These results suggested that the needle of the yew at the Ullung Island could be suitable materials as a renewable source for the mass production of taxol.
Optimum Conditions for the Protoplast Formation of Lactobacillus plantarum and Leuconostoc mesenteroides
KSBB Journal, volume 9, issue 2, 1994, Pages 191~199
Protoplasts of both strains were produced by lysozyme digestion at
for 180min. Both strains were treated with
/ml of lysozyme in 30mM Tris-HCl buffer(pH 7.5) containing 10% sucrose at the late logarithmic growth phase. It was found that the efficiency of protoplast formation was high at
and pH 7.5 by measuring the decrease in absorbance. Optimum concentrations of sucrose
for protoplast formation were determined to be 15%, 20mM and 6mM, respectively. Hydrolysis of cell wall and protoplast formation efficiency for L. plantarum showed better results than those for Leu. mesenteroides. The resistances to antibiotics erythromycin and chloramphenicols were chosen as the selection marker for the fusant between L. plantarum and Leu. mesenteroides. Production phase of protoplast in Leu. mesenteroides was also compared with L. plantarum in this paper.
Sustained Cell Growth and Improved Cyclosporin A Production Capablity of Immobilized Tolypocladium Inflatum Cells
KSBB Journal, volume 9, issue 2, 1994, Pages 200~210
In batch bioreactor fermentations for cyclosporin A (CyA) production, good potential for bioprocess improvement was demonstrated in the immobilized cell system, providing appreciably better utilization of the catalytic activity of the biomass than the freely suspended cells, especially during the exponential phase. When concentrated nutrient medium was added pulsely during the exponential phase of cell growth(at hour 139 of fermentation), reactivation and regermination in both immobilized and suspended cell cultures were observed to contribute to the longevity of CyA production, maintaining maximum CyA titre until 250 hours of fermentation. Contrarily, simple batch fermentations without any supplement of medium in both systems showed repid decrease in CyA concentrations during the late stationary phase. Notably, the CyA yield coefficient
for the immobilized cell system was maintained quite high even after the pulse addition of the concentrated full medium, reaching almost 80% of the level attained during the exponential phase. This is in sharp contrast when compared with the corresponding value of 58% in the case of parallel-suspended cells. This pattern of CyA production resulted in considerably enhanced CyA production in the immobilized cell system, reaching almost 2 time higher maximum CyA production in comparison with the free cell system.
Comparative Bioreactor Studies in Terms of Oxygen Transfer between Suspended and Immobilized Fungal Systems for Cyclosporin A Fermentation
KSBB Journal, volume 9, issue 2, 1994, Pages 211~223
In fermentations with a 4-liter stirred tank bioreactor, a better than two-fold enhancement of the gas-liquid mass transfer coefficient
in the celite-immobilized fungal cultures of Tolypocladium in flatum over the parallel conventional free-cell was observed at identical biomass concentrations, despite the higher specific oxygen uptake rate of the immobilized fungi during exponential growth. As a result oxygen sufficient conditions, i. e., dissolve oxygen(D.O.) concentrations exceeding 75% air saturation, could be maintained throughout exponential growth period of the immobilized culture, in contrast to the suspended fungal culture, whose D.O. levels fell below 50% air saturation. A linear monotonic dependence of
upon impeller agitaion rate was found for both immobilized and conventional cultivation modes over a range of 250 to 550rpm, the slope being a function of biomass concentration for the free but not for the immobilized cell system In contrasts oxygen transfer rate was a much weaker function of aeration rate up to about 2.5 vvm for both culture configurations. Above this level, aeration rate had no further effect on the mass transfer. In addition, the immobilized cultures sustained good morphological and physiological states, leading to almost two times higher cyclosporln A (CyA) productivity overt the parallel free cell system. These experiments suggest that the celite-immobilized fungal system in a stirred tank reactor has considerable promise for scaling up cyclosporin A production in terms of high-density cultivation.
Production of Antifungal Lipopeptide Iturin by Bacillus subtilis
KSBB Journal, volume 9, issue 2, 1994, Pages 224~229
Iturin, an antifungal lipopeptide, fermentation by Bacillus subtilis was investigated focusing on the effeats of nutrients aeration and specific cell growth rate on iturin production. Cell growth and product formation were not affected by different kinds of carbon sources such as sucrose, glucose and fructose. Soytone concentration above 20g/
did not influence iturin production. Diauxic growth pattern appeared when only soytone was used as a sole nitrogen source probably due to the shortage of amino acids and/or peptides in soytone which could be favorably assimilated by the cells. The composition of three major components in iturin was not changed significantly by the variation of dissolved oxygen concentration of the culture broth but changed substantially by the change of specific growth rate of the cells.
Mathematical Analysis of a High Density Animal Cell Culture with a Spin-Filter
KSBB Journal, volume 9, issue 2, 1994, Pages 230~237
Spin-filters are used as cell separation devices for achieving high cell density and high productivity in animal cell culture. We have proposed a model for the cell growth in a spin-filter perfusion culture and examined the effects on cell growth by several parameters including ammonia inhibition, specific growth rate, specific feeding rate, and cell retention. Results from computer simulation and sensitivity analysis indicate that the cell retention affects the cell growth mostly while there is a significant inhibition on cell growth by the ammonia accumulated during the culture. The specific feeding rate has minimal effects on cell growth, which is consistant with the fact that the cell growth with a step feeding is quite similar to that with a continuous feeding.