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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
Selecting the target year
Characteristics of Growth and Metal Removal in Recombinant Saccharomyces cerevisiae harboring a Metallothionein Gene
KSBB Journal, volume 10, issue 5, 1995, Pages 475~481
The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of
were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg
/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.
Comparison of Axial and Radial Flow Chromatography on Protein Separation Speed and Resolution
KSBB Journal, volume 10, issue 5, 1995, Pages 482~490
The relationship between pressure drop and liquid flow rate, for an axial and a radial flow chromatographic column packed with compressible porous media was theoretically analyzed using modified Kozeny-Carman equation. The results were compared with experimental observations obtained using compressible DEAE-agarose as a model medium. At 2-9 psi range studied, the theoretical derivation accounting for 'gel compression' effect predicted simple Langmuirian type response of volumetric flow rate to changes in pressure drop. On the other hand, the experimental response was more or less sigmoidal. At the same pressure drop, radial column showed 2-3 times higher flow rates than those of axial column both theoretically and experimentally. Using r-HBsAg crude extract, protein resolution effects between the two types of columns at different flow rates were compared side-by-side. It turned out that, though general chromatographic behavior was very similar, axial column was somewhat superior in terms of r-HBsAg recovery yield and specificity. However, the number of theoretical plates analysis indicated the protein resolution effects were comparable.
Formation of Cyclodextrin Adsorbent Using Fatty Acid as a Ligand and Fractionation of
KSBB Journal, volume 10, issue 5, 1995, Pages 491~498
In order to fraclionate
-cyclodextrins(CDs) from CD reaction mixture, various CD adsorbents were manufactured using fatty acids as the ligand molecules and anion exchange resins as matrix. Among several anion exchange resins, DEAE Cellulose was found to be the most suitable matrix for binding fatty acid. The binding stability between DEAE Cellulose and capric acid was tested under the various operation conditions, such as temperature, ethanol concentration, and ionic strength. Specific CD adsorbents manufactured with different chain-length fatty acids, saturated and unsaturated, were compared in terms of the recovery yield and selectivity of
-CDs. Stearic acid (C18, saturated) was identified as the most effective ligand for fractionation of
-CD, and linoleic acid ((C18, unsaturated ) for
-CD. The spacer length between the matrix and ligand was required for effective adsorption of CDs, and the double bond in fatty acid molecules was also acted as an important factor determining recovery yield and selectivity. The elusion patterns of
-CD from column packed with stearic acid and linoleic acid CD adsorbents were also investigated at the various elusion conditions for fractionation of
Molecular Cloning of Mutant cDNA of PU.1 Gene
KSBB Journal, volume 10, issue 5, 1995, Pages 499~509
PU.1, a tissue-specific transcription activator, binds to a purine-rich sequence(5'-GAGGAA-3') called PU box. The PU.1 cDNA consists of an open reading frame of 816 nucleotides coding for 272 amino acids. The amino terminal end is highly acidic, while the carboxyl terminal end is highly basic. Transcriptional activation domain is located at the amino terminal end, while DNA binding domain is located at the carboxyl terminal end. Activation of PU.1 transcription factor is supposed to be accomplished by the phosphorylation of serine residue(s). There exist 22 serines in the PU.1. Five(the 41, 45, 132
133, and 148th) of the serines(plausible phosphorylation site by casein kinase II), are the primary targets of interest in elucidating the molecular mechanism(s) of the action of the PU.1 gene. In this study, PU.1 cDNA coding for the five serine residues(41th AGC, 45th AGC, 132
TCA, and 148th TCT), was mutated to alanine codon(41th GCC, 45th GCC, 132
GCA, and 1481h GCT), respectively, by Splicing-Overlapping-Extension(SOE) using Polymerase Chain Reaction(PCR). And each mutated cDNA fragments was ligated into pBluescript KS＋ digested with HindIII and Xba I, to generate mutant clones named pKKS41A, pRKS45A, pMKS132
133A, and pMKS148A. The clones will be informative to study the "Structure and Function" of the immu-nologically important gene, PU.1.
Development of Natural Seasoning from Alaska Pollack Skin Gelatin Using Continuous Three-Step Membrane Reactor
KSBB Journal, volume 10, issue 5, 1995, Pages 510~517
The hydrolysates of three kinds [FSEH(first step enzymatic hydrolysate), SSEH(second step enzymatic hydrolysate), and TSEH(third step enzymatic hydyolysate)] were prepared by continuous hydrolysis of Alaska pollack(Theragra chalcogramma) skin gelatin with three-step membrane enzyme reactor. The molecular weight distributions of FSEH, SSEH, and THSE are 9,500∼4,800Da, 6,600∼3,400Da, and 2,300∼900Da, respectively. The contents of amino acid having sweet taste (glycine, proline, serine, alanine, hydroxyproline, glutamic acid, and aspartic acid) were about 70% of total amino acid being in the three kind hydrolysates. We also tried preparing of natural seasonings (complex seasoning and enzymeatic hydrolysale sauce) using the hydrolysates. From the results of sensory evaluations, complex seasoning containing TSEH was nearly equal to shellfish complex seasoning on the market. The mixture sauce which was made by mixing of 80% enzymatic hydrolysis sauce and 20% fermented soy sauce, was at least similar to the tradition soybean sauce in product quality, too.
Production Conditions of Two Polysaccharides from Marine Bacterium Zoogloea sp.
KSBB Journal, volume 10, issue 5, 1995, Pages 518~524
Marine bacterium, as a microbial source producing polysaccharides, was newly isolated from the eastern and western sea of Korea and was identified as Zoogloea sp. (KCCM 10036). It produced two different types of polysaccharides, especially: WSP (water-soluble polysaccharide) and CBP (cell-bound polysaccharide). The former was isolated from the supernatant of centrifuged broth by acetone precipitation, and the latter was isolated from the pellet by acetone and CPC (cetylpyrldinium chloride) precipitation. The productivity of polysaccharides were increased with the addition of promoting agents such as biotin, ampicillin and surfactant. After batch fermenting, the productivity of WSP and CBP were reached to maximum values of
in the culture medium containing 1% of glucose as a carbon source.
Inhibitory Effects of Tumor Metastasis by Chitosan Derivative, of Sulfated N-acetyl Chitosan
KSBB Journal, volume 10, issue 5, 1995, Pages 525~532
Chitosan derivative, of a sulfated N-acetyl chitosan was synthesized, and the inhibitory effects of this compound on the experimental and spontaneous lung metastallc B16/BL6 melanoma bearing mice were investigated. Position of substitution with sulfate in water-soluble sulfated derivatives of chitosan were analysed by 13C-nmr. The structure of N-acetyl chitosan 3,6 0-disulfate were confirmed. The tumor growth inhibition of B16/BL6 melanoma cells has been shown at the highest level of 77.6% when sulfated N-acetyl chitosan were administered at the dose of 100mg/kg. In the lung metastasls, the sulfated N-atetyl chitosan was administered to C57BL/6B mice bearing B16/BL6 melanoma cells by I.V. injection and the number of metastasis foci of melanoma were decreased by the dose dependent manner ranging from 20 to 100mg/kg. In the spontaneous metastasis, I.V. administrations of sulfated N-acetyl chitosan after tumor inoculation resulted in marked reduction of metastatic colonies. A sulfated N-acetyl chitosan was able to partially inhibit the tumor cell adhesion by migration to laminin. These results suggested that chitosan derivative, a sulfated N-acetyl chitoasn was able to inhibit to the experimental and spontaneous metastasis models as well as cell adhesion ability.
Recovery of Poly(3-hydroxybutyrate) from Recombinant Escherichia coli by Autolysis with Bacteriophage Lambda
KSBB Journal, volume 10, issue 5, 1995, Pages 533~539
An autolytic system based on a thermally inducible phage lambda, λHL1, has been applied for the recovery of poly(3-hydroxybutyrate) [PHB] from a recombinant Escherichia coli XL1-Blue, harbouring a plasmid (pSYL105) containing the Alcaligenes eutrophus PHB biosynthesis genes. The lytic capability ofλHL1 was evaluated in flask culture for both lysogens, XL1-Blue (λHL1) and XL1-Blue (λHL1, pSYL105). When the optical density of culture at 600nm(OD600) reached 0.2, cell lysis was induced by increasing the temperature from
. Most cells of XL1-Blue (
HL1) were lysed by the autolytic system in an hour after the thermal induction, while the lytic efficiency was slightly lower for XLl-Blue (λHL1, pSYL105). The existence of pSYL105 in cells seemed to inhibit, to some extent, the lytic capability of λHL1 even at low PHB content. The lylic efficiency remarkably decreased as the induction was delayed to allow PHB accumulation. When a chemical induction using 2% (v/v) chloroform was introduced after an hours of thermal induction, we could obtain a good lytic efficiency.
Reaction Characteristics and Kinetic Analysis of Enzymatic Hydrolysis of Corn Gluten Meal Using Alkaline Protease
KSBB Journal, volume 10, issue 5, 1995, Pages 540~546
Dry corn gluten meal of 70% protein content was enzymatically hydrolyzed by alkaline protease in a pH-state reactor. Such process variables as temperature, pH, and enzyme-to-substrate ratio were varied, and at each condition degree of hydrolysis was monitored and calculated. The ultimate degree of hydrolysis, which ranged between 25 and 28% based on gluten protein mass, was not significantly affected by the process variables. However,
and pH 9-10 appeared optimum. Kinetic analysis indicated enzyme deactivation was negligible during the hydrolysis, and the experimental data were near perfectly fitted to the model kinetic equation which was modified after neglecting enzyme deactivation term. The enzyme reaction was 1
scaled up and basically the same hydrolysis performance was resulted. Amino acid analysis showed the hydrolyzate was relatively rich in glutamine/glutamic acid, leucine, and alanine at 19.6, 16.1, and 12.3 mole %, respectively.
Functional and Rheological Properties of
-Polyglutamic Acid Produced by Alkalophilic Alcaligenes sp.
KSBB Journal, volume 10, issue 5, 1995, Pages 547~552
Some functional and rheological properties of the
-PGA) produced from alkalophilic Alcaligenes sp. were investigated. Viscosity synergism with thickening agents, capacities for gelling, entrapig of heavy metals, and flocculability of
-PGA were not observed, but the relatively good compatability with various polyvalent metallic ins, excellent absorption ability and spinability were observed. The
-PGA solution showed non-Newtonian flow behavior and exhibited pseudoplastic property with a yield stress at above 1% concentration. The values of flow index for 1% solution were in the range of 0.41∼0.75 showing shear rate dependency and the value of yield stress was 2.28 Pa. The value of consistency index was 0.868 Pa
Sn and was exponentially dependent on concentration.
Antibacterial Activity of actobacillus sp. KJ-5 Isolated from Pig Feces
KSBB Journal, volume 10, issue 5, 1995, Pages 553~560
A lactic aci bacteria producing antibacterial substance was isolated from pig feces. This strain was identified as a genus Lactobacillus, through its morphological, cultural and physiological characteristics. Lactobacillus sp. KJ-5 isolated showed the strong inhibitory effect on the growth of Salmonella paratyphi. The production of antibacterial substance was growth associated form during the batch culture of Lactobacillus sp. KJ-5 and the maximum production was obtained at the culture temperature of
as well as optimum temperature of cell growth. The antibacterial activity of the filtrate of culture broth was decreased by adjusting the pH 6.2 and was not affected by catalase treatment. The antibacterial substance was partially purified by methanol and acetone extraction, whtch exhibited three spots in the thin-layer chromatography and one of them showed an antibacterial activity, This substance also showed the maximum absorption of UV at 270nm and an antibacterial activity was completely inactivated by the treatment of proteolytic enzymes.
Oxygen Toxicity of Superoxide Dismutase-Deficient Saccharomyces cerevisiae by Paraquat
KSBB Journal, volume 10, issue 5, 1995, Pages 561~567
Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.
Effect of pH on Organic Acid Production by Anaerobiospirillum succiniciproducens
KSBB Journal, volume 10, issue 5, 1995, Pages 568~574
To investigate the effect of pH on organic acid production by Anaerobiospirillum succiniciproducens, an anaerobic fermentation was carried out by maintaining the pH of the fermentation broth at 5.8, 6.0, 6.4, 6.8, and 7.2. At various pHs, the concentrations of cell were
which were two to three times higher than those of the other worker's results, and the maximum was obtained at pH 5.8. Substrate consumption was increased by increasing the pH in the range of pH 6.0 to 6.8, while the sugar consumption rate at both pH 5.8 and 7.2 was very slow. The total amount of 2M
added for adjustment of pH change due to organic acid production was maximum at pH 6.8. Changes of conductivity of the fermentation broth was very simillar to those of 2M
added at various pHs. Therefore, it is suggested that determination of the amount of organic acid in a broth can be possible by measuring the conductivity. The maximum production yield of lactate based on glucose was 64% for pH 7.2 and 32% for pH 6.8, respectively.
Controlled Lysis of Lipase-Producing Recombinant E. coli by Phage Induction
KSBB Journal, volume 10, issue 5, 1995, Pages 575~581
A plasmid pTTY2, containing the lipase-producing gene, was used to transform an E. coli phage lysogen, P90c/
434, into the lipase-producing lysogen, P90c/
434/pTTY2. After the overproduction of lipase by the isopropylthio-
-D-galactoside induction, the prophage
434 in the chromosome of the host cell was induced by the milomycin C addition or ultraviolet irradiation to lyse the host cell. The optimum operating conditions, such as the isopropylthio-
-D-galactoside induction period and the phage induction timing, were sought for the efficient cell lysis in the same fermenter. Effective cell lysis occurred at the earlier exponential growth phase with the isopropylthio-
-D-galactoside induction period of 1 hour. The amount of the lipase production was qualitatively measured by the halo size in Luria-Bertani agar medium containing tributyrin and Rhodamine B plate.
Purification and Characterizationn of Biosurfactant from Marine Pseudomonas sp. CHCS-2
KSBB Journal, volume 10, issue 5, 1995, Pages 582~588
A marine microorganism producing biosurfactant was isolated from the oil polluted coast of Chung-Mu in Korea, and was identified as Pseudomonas sp.. It produced the biosurfactanl and its optimum culture conditions for pH and salt concentration were 8.0 and 3.0%, respectively. The productivity of biosurfactant from this strain was affected by the nitrogen source used. For the oil resolvability of the biosurfactant, the residual oil in the culture broth with 2% Kuwait crude oil at each time of 48, 96, and 132hr was investigated by gas chromatography. As result of this experiment, it was verified that the biosurfactant acted on C10-C14, of Kuwait crude oil and so the oil was decomposed. The biosurfactant isolated from the supernatant was purified by adsorption to Amberliter XAD-7 and followed by gel chromatography (Sephadex G-100) and HPLC. The purified biosurfactant showed a high value of emulsifying activity at
and the emulsifying stability was maintained at the temperature range of
. The purified biosurfactant reduced the interfacial tension of Kuwait crude oil remarkably and showed improved dispersing ability compared to those of commercial surfactants such as Tween 80, Tween 60 and SDS.
Isolation and Purification of Protein-bound Polysaccharides from Mycelia of Flammulina velutipes Grown on Sawdust Medium
KSBB Journal, volume 10, issue 5, 1995, Pages 589~597
Protein-bound polysaccahrides(PBP) were isolated, purified, and characterized from the sawdust media after harvesting the fruit body of Flammulina velutipes. The yield of the crude PBP(Fr.CA) extracted from the sawdust media, was 0.367% relative to the original sawdust media. The total sugar and protein contents of Fr.CA were 19.8% and 23.8% respectively. Using the membrane filtration, the fraction of which the molecular weight is over 300 kDa(Fr.A) was isolated from the Fr.CA and the yield was 44.6% relative to the Fr.CA. This result indicates that high molecular PBP is the dominant components of the Fr.CA. The Fr.A was separated into three fractions (Fr.A-1, Fr.A-2 and Fr.A-3) whose yields are 5.8%, 8.5% and 13.2% respectively. These fractions were further purified using gel filtration, obtaining a single peak in each fraction that considered as pure PBP Among them, the yield of Fr.A-1-
was 20.9% relative to the Fr.A-1, and the molecular weight was 800 kDa. Monosaccharide components such as glucose, galactose, mannose, xylose and fucose could be detected all fractions by a HPLC analysis. Especially in the Fr.A-1-
fraction, the content of glucose and galactose appeared to be high.
Ethanol Production from Tapioca Hydrolysate by Batch and Continuous Cell Retention Cultures
KSBB Journal, volume 10, issue 5, 1995, Pages 598~603
Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/
was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/
making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5
107 cells/ml(about 2.1g/
). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/
, about 15g/
, and 14.7g/
-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/
and the ethanol concentration oscillated around 68g/
. The volumetric ethanol productivity was about 13.6g/
-h and the residual reducing sugar concentration about 12g/
containing glucose of about 4.5g/
. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.