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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Microbiology and Biotechnology
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 12, Issue 6 - Dec 2002
Volume 12, Issue 5 - Oct 2002
Volume 12, Issue 4 - Aug 2002
Volume 12, Issue 3 - Jun 2002
Volume 12, Issue 2 - Apr 2002
Volume 12, Issue 1 - Feb 2002
Selecting the target year
Development of Medium for Griseofulvin Production: Part I. Screening of Medium Constituents Using the Plackett-Burman Experimental Design
Venkata, Dasu-V. ; Panda, T. ; Chidambaram, M. ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 355~359
The Plackett-Burman experimental design was employed to evaluate the relative importance of medium constituents of each medium for enhanced griseofulvin production by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004. It was found that the medium constituents, sucrose,
, significantly influenced the griseofulvin production by Penicillium griseofulvum MTCC 1898. In the case of Penicillium griseofulvum MTCC 2004, lactose, glucose, and
significantly influenced the griseofulvin production.
Development of Medium for Griseofulvin Production: Part II. Optimization of Medium Constituents Using Central Composite Design
Dasu, Venkata ; Panda, V.T. ; Chidambaram, M. ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 360~366
Central composite experimental design was employed to determine the optimal concentration of medium constituents for griseofulvin production by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004. The optimal concentration of sucrose,
were found to be 48.08 g/1, 1.228 g/1 , 2.7 g/1, and 0.011 g/1, respectively, for Penicillium griseofulvum MTCC 1898, and for Penicillium griseofulvum MTCC 2004, 23.52 g/1, 43.67 g/1, and 0.0434 g/1 of glucose, lactose, and
, respectively. The yield of griseofulvin under optimal composition of medium constituents increased by 1.26 and 1.38 times than prior to optimization, for Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvutn MTCC 2004, respectively.
The Hepatoprotective Effects of Polysaccharides Isolated from Submerged Fermentation of Ganoderma lucidum
Zhang, Xue-Hong ; Hu, Hong-Bo ; Tang, Yong-Lian ; Huang, Rui-Shan ; Luo, Jiu-Fu ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 367~370
A neutral polysaccharide, GP, was isolated from a fermentation broth of Ganoderma lucidum Acid hydrolysis and a paper chromatography analysis indicated that the polysacchride was composed of glucose, xylose, and mannose. The molecular weight was estimated to be
. The oral administration of GP to mice showed that it can inhibit liver damage induced by GalN and
Glycolipid Biosurfactants Produced by Pseudomonas aeruginosa D2D2 from Diesel-Contaminated Soil
MOON, HYE-JOON ; YOUNG-KUONG LIM ; HEE-SIK KIM ; DAE-YOUNG KWON ; WOOK-JIN CHUNG ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 371~376
A biosurfactant-producing bacterial strain was selected from diesel-contaminated soil by measuring the oil-film collapsing activity and identified as Pseudomonas aeruginosa D2D2. When glucose and olive oil were used as carbon sources, 11.46 g/1 of biosurfactant was obtained. Based on TLC analysis, the biosurfactant produced from P. aeruginosa D2D2 was identified as a glycolipid, consisting of two types of biosurfactants (Type I and Type II). The purified glycolipid reduced the surface tension of the culture from 72 dyne/cm to 27 dyne/cm. The hydrophilic and hydrophobic moiety of the biosurfactant were rhamnose and
-hydroxydecanoic acid, as determined by FAB-MS and NMR analyses, respectively.
Estimation of Nitrite Concentration in the Biological Nitritation Process Using Enzymatic Inhibition Kinetics
GIL, KYUNG-IK ; EUI-SO CHOI ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 377~381
Recently, interests to remove nitrogen in the nitritation process have increased because of its economical advantages, since it could be a short-cut process to save both oxygen for nitrification and carbon for denitrification compared to a typical nitrification. However, the kinetics related with the nitritation process has not yet been fully understood. Furthermore, many useful models which have been successfully used for wastewater treatment processes cannot be used to estimate effluent nitrite concentration for evaluating performance of the nitritation process, since the process rate equations and population of microorganisms for nitrogen removal in these models have been set up only for the condition of full nitrification. Therefore, the present study was conducted to estimate an effluent nitrite concentration in the nitritation process with a concept of enzymatic inhibition kinetics based on long-term laboratory experiments. Using a nonlinear least squares regression method, kinetic parameters were accurately determined. By setting up a process rate equation along with a mass balance equation of the nitrite-oxidizing step, an effluent nitrite concentration in the nitritation process was then successfully estimated.
FAME Analysis to Monitor Impact of Organic Matter on Soil Bacterial Populations
Kim, Jong-Shik ; Joo, Jin-Bee ; Weon, Hang-Yeon ; Kang, Chang-Seong ; Lee, Si-Kyung ; Yahng, Chahng-Sool ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 382~388
In order to assess the effects of organic fertilizer on soil microbial community structure and diversity in the greenhouse fields, fatty acid methyl ester (FAME) was analyzed by the MIDI (Microbial ID, Inc., Newark, DE, U.S.A.) system and enumerations were performed. In relation to bacterial division of each sample, low GC Gram-positive bacteria were predominant among bacteria cultured on aerobic bacteria media. On the other hand, alpha subdivision was predominant on proteobacteria of control and OM (organic matter) 1 treated plot, and Flavobacterium spp. existed in OM2 plot on crystal violet media of all samples. Shannon-weaver Index (H) of OM1 plot varied most by 1.9 and 5.0 among bacteria cultured on aerobic bacteria media and crystal violet media, respectively. Our results revealed that addition of the organic wastes to soil led to a highly diverse microbial community, but the excessive amounts of organic and mineral fertilizer applied in the greenhouse fields produced excess nutrients in soil and led to simplification on bacterial populations.
Isolation of a Nisin-Producing Lactococcus lactis Strain from Kimchi and Characterization of its nisZ Gene
Lee, Kwang-Hee ; Moon, Gi-Seong ; An, Jong-Yun ; Lee, Hyong-Joo ; Chang, Hae-Choon ; Chung, Dae-Kyun ; Lee, Jong-Hoon ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 389~397
Bacteriocin-producing lactic acid bacteria were isolated from kimchi. One isolate producing the most efficient bacteriocin was identified and named Lactococcus lactis B2, based on the biochemical properties and 16S rDNA sequences. The B2 bacteriocin inhibited many different Gram positive bacteria including Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Streptococcus, and Staphylococcus, but did not inhibit Gram-negative bacteria. The bacteriocin was maximally produced at temperatures between
and at the initial pH of 7.0. Ninety
of the activity remained after 10 min of heat treatment at
, after 1 h exposure to organic solvents. The bacteriocin was purified from culture supernatant by ammonium sulfate precipitation, CM Sepharose column chromatography, ultrafiltration, and finally, by reverse-phase HPLC. A 1.58-kb fragment was amplified from B2 chromosome by using a primer set designed from the published nisA sequence. Sequencing result showed that the fragment contained the whole nisZ and 5' portion of nisB, whose gene product was involved in postmodification of nisin. The upstream sequence, however, was completely different from those of reported nisin genes.
Cytoskeleton Reorganization and Cytokine Production of Macrophages by Bifidobacterial Cells and Cell-Free Extracts
Lee, Myung-Ja ; Zang, Zhen-Ling ; Choi, Eui-Yul ; Shin, Hyun-Kyung ; Ji, Geun-Eog ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 398~405
Bifidobacteria have been previously shown to stimulate the immune functions and cytokine production in macrophages and T-lymphocytes. Accordingly, the RAW 264.7 murine macrophage cell line was used to assess the effects of Bifidobacterium on the proliferation and cytoskeleton reorganization of the cells. Cytokine production after exposure to Bifidobacterium was also monitored in both whole cells and cell-free extracts. When RAW 264.7 cells were cultured for 24 h in the presence of heat-killed Bifidobacterium bifidum BGN4, the proliferation of macrophages was slowed down in a dose-dependent manner and cell differentiation was observed by staining with the actin-specific fluorescent dye, rhodamin-conjugated phalloidin. Although EL-4 cells, a T-cell line, stimulated RAW 264.7 cells to produce TNF-
and IL-6, the stimulatory activity of B. bifidum BGN4 decreased as the EL-4 cell number increased. When disrupted and fractionated BGN4 was used, the whole cell fraction was more effective than the other fractions for the TNF-
production. In contrast, the cell-free extract exhibited the highest IL-6 production level among the fractions, which was evident even at a
concentration. The current results demonstrate that Bifidobacterium induced differentiation of the macrophages from the fast proliferative stage and that the cytokine production was differentially induced by the whole cells and cell-free extracts. The in vitro approaches employed herein are expected to be useful in further characterization of the effects of bifidobacteria with regards to gastrointestinal and systemic immunity.
A New Spin Filter for High Density Culture and Ethanol Production by Saccharomyces cerevisiae
Moon, Hyun-Soo ; Lim, Dong-Joon ; Song, Gu-Young ; Kang, Hyun-Ah ; Kim, Seung-Wook ; Kim, Ik-Hwan ; Hong, Suk-In ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 406~410
A new spin filter consisting of
(nominal pore size) depth fitters rolled on a stainless steel grid was developed, using Saccharomyces cerevisiae as a model suspension cell to evaluate the spin filter performance. In a 1.8-1 fermentor with a rotation speed of 300 rpm and perfusion rate of 4 ml/min, a cell concentration of 49 g/l and ethanol concentration of 45 g/l from 100 g/l glucose could be obtained in a perfusion culture. The major mechanisms for cell separation used by the large-pore spin filter appeared to be centrifugal force and pivotal movement of the cells in the spin filter.
Display of Bacillus macerans Cyclodextrin Glucanotransferase on Cell Surface of Saccharomyces cerevisiae
Kim, Kyu-Yong ; Kim, Myoun-Dong ; Han, Nam-Soo ; Seo, Jin-Ho ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 411~416
Bacillus macerans cyclodextrin glucanotransferase (CGTase) was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane-anchored protein, Aga1p. The surface display of CGTase was confirmed by immunofluorescence microscopy and its enzymatic ability to form
-cyclodextrin from starch. The maximum surface-display of CGTase was obtained by growing recombinant S. cerevisiae at
and pH 6.0. S. cerevisiae cells displaying CGTase on their surface consumed glucose and maltose, inhibitory byproducts of the CGTase reaction, to enhance the purity of produced cyclodextrins. Accordingly, the experimental results described herein suggest a possibility of using the recombinant S.cerevisiae anchored with bacterial CGTase on the cell surface as a whole-cell biocatalyst for the production of cyclodextrin.
Detection of Aromatic Pollutants by Bacterial Biosensors Bearing Gene Fusions Constructed with the dnaK Promoter of Pseudomonas sp. DJ-12
Park, Sang-Ho ; Lee, Dong-Hun ; Oh, Kye-Heon ; Lee, Kyoung ; Kim, Chi-Kyung ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 417~422
Gene fusions were constructed by the transcriptional fusion of the dnaK promoter of pseudomonas sp. DJ-12 or E. coli to the lux or luc marker gene. The dnaKp-DJ::luxCDABE bioluminescent fusion in the biosensor using the Pseudomonas sp. DJ-12 dnaK promoter exhibited about 5-fold more extensive response to ethanol than that of dnaKp-EC::luxCDABE. The bioluminescent response of the dnaK-DJ::luc fusion to ethanol was much weaker than those of the other fusions. The biosensor harboring the dnaKp-DJ::luCDABE fusion was examined for its bioluminescence production based on exposure to aromatic compounds, such as biphenyl, 4-chlorobiphenyl (4CB), 4-hydroxybenzoate (4HBA), and catechol. In particular, the bioluminescence produced by the dnaKp-DJ::luxCDABE fusion was most sensitive to 1 mM biphenyl and 4CB when exposed for 80 min, and the responses were also very strong to other aromatics. Therefore, the biosensor bearing the dnaKp-DJ::luxCDABE fusion would appear to be the most useful for the detection of aromatics and other pollutants.
Interactive Effects of UV-B and Pesticides on Photosynthesis and Nitrogen Fixation of Anabaena doliolum
Chandrai, Lar ; Vandana, Pandey ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 423~430
The effects of UV-B and commercial grade pesticides (butachlor and carbofuran), individually and in combination, were studied on a variety of physiological processes of rice field cyanobacterium Anabaena doliolum. Butachlor was found to be
more toxic than carbofuran and
than UV-B on the growth, photosynthesis, lipid peroxidation, membrane permeability, and nitrogenase activity of the test cyanobacterium. Of the three photosynthesis inhibitors, the butachlor-induced inhibition of whole chain was approximately 3 and
higher than carbofuran and UV-B, respectively. Although the interaction of the stress factors caused a significant inhibition (P<0.01), it was still less than the additive effect on the parameters investigated, except for PSI.
Evaluation of Fertilizer Additions to Stimulate Oil Biodegradation in Sand Seashore Mesocosms
CHOI, SUNG-CHAN ; KAE KYOUNG KWON ; JAE HAK SOHN ; SANG-JIN KIM ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 431~436
Effects of fertilizer additions for oil degradation were examined in sand seashore mesocosms. Within 37 days, up to
removal was achieved by the addition of slow-release type fertilizer (SRF) with the initial degradation rate of 423.3 mg oil
. The removal was mostly of biological origin based on the changes of
/phytane ratios from 2.60 to 0.81 and from 3.55 to 1.29, respectively. The addition of oleophilic fertilizer (Inipol EAP22) was less effective and resulted in the removal of
of the added oil (
, v/v) with a lower initial degradation rate. Petroleum-degrading bacteria had achieved a value of
at Day 3 and this peak exactly coincided with the initial degradation in the SRF-treated mesocosm. In this mesocosm, surface tension values were decreased drastically during Days 3 and 8, suggesting that microbially-produced surface-active agents actively enhanced the oil degradation rate and cell proliferation. Although the Inipol-treated mesocosm appeared to show significantly enhanced oil degradation compared to that of the untreated control mesocosm, Inipol was found to be less effective than SRF in enhancing a true oil-degrader when compared under similar experimental conditions.
Identification of Stenotrophomonas maltophilia LK-24 and its Degradability of Crystal Violet
Kim, Jeong-Dong ; Yoon, Jung-Hoon ; Park, Yong-Ha ; Fusako Kawai ; Kim, Hyun-Tae ; Lee, Dae-Weon ; Kang, Kook-Hee ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 437~443
A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a
homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below
, yet the growth of the cells was totally suppressed at a dye concentration of
. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and
-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the
-dimethylaminophenol was not easily identifiable, as it was further metabolized.
High Cell Density Cultivation of Bifidobacterium longum Using a Calcium Carbonate-Alginate Beads System
Yu, Won-Kyu ; Kim, Ji-Youn ; Lee, Ki-Yong ; Heo, Tae-Ryeon ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 444~448
-alginate beads system was developed for high cell density cultivation of Bifidobacterium longum and the cost-effective media were also screened. In batch process with
, beads, two strains of B. longum showed both the highest viable cells and optical density in TPY medium, resulting in maximum optical density and viable cell counts of 12.40,
cfu/ml for B. longum ATCC 15707 and 13.71,
cfu/ml for B. longum HLC 3742. Released size distribution, according to
-alginate bead size preparation, was smaller than others. These results were also examined by observing their morphology. The skim milk-based medium was most adequate to cultivate B. longum as the cheapest medium, and
skim milk supplemented with
yeast extract was a suitable medium, supporting the growth to
cfu/ml for ATCC 15707 and
cfu/ml for HLC 3742. During the long-term storage at
, B. longum cultivated with
beads had the highest stability. Consequently,
-alginate beads buffer was found to be useful not only to cultivate B. longum but also to preserve cultures.
New Response Surface Approach to Optimize Medium Composition for Production of Bacteriocin by Lactobacillus acidophilus ATCC 4356
RHEEM, SUNGSUE ; SEJONG OH ; KYOUNG SIK HAN ; JEE YOUNG IMM ; SAEHUN KIM ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 449~456
The objective of this study was to optimize medium composition of initial pH, tryptone, glucose, yeast extract, and mineral mixture for production of bacteriocin by Lactobacillus acidophilus ATCC 4356, using response surface methodology. A response surface approach including new statistical and plotting methods was employed for design and analysis of the experiment. An interiorly augmented central composite design was used as an experimental design. A normal-distribution log-link generalized linear model based on a subset fourth-order polynomial (
=0.94, Mean Error Deviance=0.0065) was used as an analysis model. This model was statistically superior to the full second-order polynomial-based generalized linear model (
=0.80, Mean Error Deviance=0.0140). Nonlinear programming determined the optimum composition of the medium as initial pH 6.35, typtone
, yeast extract
, and mineral mixture
. A validation experiment confirmed that the optimized medium was comparable to the MRS medium in bacteriocin production, having the advantage of economy and practicality.
Enhancement of Tissue Type Plasminogen Activator (tPA) Production from Recombinant CHO Cells by Low Electromagnetic Fields
Lee, Seo-Ho ; Lee, Hyun-Soo ; Lee, Mi-Kyoung ; Lee, Jin-Ha ; Kim, Jong-Dai ; Park, Young-Shik ; Lee, Shin-Young ; Lee, Hyeon-Yong ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 457~462
Low Electromagnetic Field (EMF) intensity in the range of
(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At
viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to
viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain
viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between
in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at
EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of
was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than
could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.
Expression of Rotavirus Capsid Proteins VP6 and VP7 in Mammalian Cells Using Semliki Forest Virus-Based Expression System
Choi, Eun-Ah ; Kim, Eun ; Oh, Yoon-I ; Shin, Kwang-Soon ; Kim, Hyun-Soo ; Kim, Chul-Joong ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 463~469
Rotaviruses are the world-wide leading causative agents of severe dehydrating gastroenteritis in young children and animals. The outer capsid glycoprotein VP7 and inner capsid glycoprotein VP6 of rotaviruses are highly antigenic and immunogenic. An SFV-based expression system has recently emerged as a useful tool for heterologous protein production in mammalian cells, exhibiting a much more efficient performance compared to other gene expression systems. Accordingly, the current study adopted an SFV-based expression system to express the VP7 of a group A human rotavirus from a Korean isolate, and the VP6 of a group B bovine rotavirus from a Korean isolate, in mammalian cells. The genes of the VP6 and VP7 were inserted into the SFV expression vector pSFV-1. The RNA was transcribed in vitro from pSFV-VP6 and pSFV-VP7 using SP6 polymerase. Each RNA was then electroporated into BHK-21 cells along with pSFV-helper RNA containing the structural protein gene without the packaging signal. The expression of VP6 and VP7 in the cytoplasm was then detected by immunocytochemistry. The recombinant virus was harvested by ultracentrifugation and examined under electron microscopy. After infecting BHK-21 cells with the defective viruses, the expressed proteins were separated by SDS-PAGE and analyzed by a Western blot. The results indicate that an SFV-based expression system fur the VP6 and VP7 of rotaviruses is an efficient tool for developing a diagnostic kit and/or preventive vaccine.
Isolation and Biological Properties of Novel Cell Cycle Inhibitor, HY558, Isolated from Penicillium minioluteum F558
Lee, Chul-Hoon ; Lim, Hae-Young ; Kim, Min-Kyoung ; Cho, Youl-Hee ; Oh, Deok-Kun ; Kim, Chang-Jin ; Lim, Yoon-Gho ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 470~475
In the course of screening for a novel cell cycle inhibitor, a potent Cdk 1 inhibitor, HY558, was found from the culture broth of Penicillium minioluteum F558 isolated from a soil sample. The molecular ion of HY558 was identified at m/z 329 (MH+) with a molecular formula of
. HY558 exhibited selective antiproliferative effects on various human cancer cell lines. Its
values were estimated to be 0.29 mM on HepG2, 0.30 mM on HeLa, 0.30 mM on HL6O, 0.33 mM on HT-29, and 0.25 mM on AGS cells. Interestingly, Hy558 demonstrated no antiproliferative effect with normal lymphocytes used as the control, and a low level of inhibition on the proliferation of A549 cancer cells. A flow cytometric analysis of HepG2 cells revealed an appreciable arrest of cells at the G1 and G2/M phases of the cell cycle following treatment with Hy558. furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with 0.46 mM of HY558.
Properties of Electron Carriers in the Process of Methanol Oxidation in a New Restricted Facultative Marine Methylotrophic Bacterium, Methylophaga sp. MP
Koh, Moon-Joo ; Kim, Chun-Sung ; Kim, Yun-A ; Choi, Hack-Sun ; Cho, Eun-Hee ; Kim, Eung-Bin ; Kim, Young-Min ; Kim, Si-Wouk ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 476~482
Methanol dehydrogenase (MDH) and c-type cytochromes from marine methanol-oxidizing bacterium, Methylophaga sp. MP, were purified and characterized. The native MDH had a molecular mass of 148 kDa and its isoelectric point was 5.5. Two c-type cytochromes,
, were found, and their isoelectric points were 3.4 and 8.0, respectively. The purified MDH had higher thermal stability than that of the other soil methylotrophic bacteria. The electron flow rate from MDH to cytochrome
was higher than that from MDH to cytochrome
, indicating that the physiological primary electron acceptor for MDH is cytochrome
. The electron transfer from MDH to phenazine ethosulfate (PES, artificial electron acceptor) in the two dye (PES/DCPIP)-linked assay system was not inhibited by NaCl, whereas the electron flow from MDH to cytochrome
in the cytochrome/DCPIP-linked assay system was suppressed significantly by NaCl. Metal chelating agents such as EDTA showed the same effects on the MDH activity.
12 and G
13 Subunits Modulate
-Induced Histamine Release in Human Umbilical Cord Blood-Derived Mast Cells
Ro, Jai-Youl ; Kim, Ji-Young ; Ha, Ji-Hee ; Lee, Chang-Ho ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 483~489
The role of
in modulating the IgE receptor-mediated histamine secretion in the streptolysin-o-permeabilized human cultured mast cell was investigated. The expression of
proteins were regulated during human cultured mast cell differentiation, and a significant correlation was observed between the levels of expression of
proteins and IgE receptor-mediated histamine secretion capability in human cultured mast cells. Antibodies against
effectively inhibited the IgE receptor-induced histamine release, and the concentration of anti-
antibody used to inhibit histamine secretion was shown to also inhibit the IgE receptor-mediated elevation of intracellular
. Therefore, the results suggest that
play roles in modulating IgE receptor-activated
influx, thereby regulating histamine release in cultured human mast cells. This is the first report to show that
are involved in the regulation of
mediated exocytosis in human cultured mast cells.
Medium Optimization for Phytase Production by Recombinant Escherichia coli Using Statistical Experimental Design
Choi, Won-Chan ; Oh, Byng-Chul ; Kim, Hyung-Kwoun ; Lee, Eun-Sook ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 490~496
The production of E. coli WC7 phytase from a recombinant E. coli strain was optimized using a statistical experimental design approach. Two-level complete factorial designs with seven variables were used for the media optimization. In the first optimization step, the influence of disodium succinate, yeast extract,
, NaCl, and trace elements on phytase production was evaluated. As a result, disodium succinate, yeast extract,
, NaCl, and the trace elements were found to have a positive influence on the phytase production, while
had a negative influence. In the second step, the concentrations of disodium succinate and yeast extract were further optimized using central composite designs. The maximum phytase activity obtained was 234 U/ml using 15.9 g/1 disodium succinate, 20 g/1 yeast extract, 5 g/1
, 4 g/1 NaCl, and 1.5 m1/1 trace elements, which was about a 14-fold increase in comparison with that obtained using the basal medium.
Diversity of Leuconostocs on Garlic Surface, an Extreme Environment
KIM, MYUNG HEE ; SUN TAEK SHIM ; YOUN SOON KIM ; KYU HANG KYUNG ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 497~502
Thirty-nine strains of Leuconostocs found to be tolerant to
or more garlic were selected for further identification, by comparing their whole-cell protein pattern, 16S rRNA gene (first 530 bases) sequence, cellular fatty acid composition, and carbon source metabolism. Two isolates were Identified as Leuconostoc mesenteroides and 32 others as Leuconostoc citreum. Five other strains belonging to a cluster could not be allocated to the existing species. 16S rRNA gene sequence and cellular fatty acid composition of the unidentified bacteria exhibited close similarity with Leuconostoc argentinum. The unidentified isolates were not allocated to L. argentinum, because they formed polysaccharide from sucrose, while L. argentinum strains do not. Leuconostocs tolerant to high concentration of garlic were found predominantly on garlic surface, an extreme environment which is unfit for most of other microorganisms.
Antitumor Activity of Chitosan Oligosaccharides Produced in Ultrafiltration Membrane Reactor System
Jeon, You-Jin ; Kim, Se-Kwon ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 503~507
Chitosan oligosaccharides (COSs) were prepared and fractionated into three groups of COS [a high molecular weight COS (HMWCOS), medium molecular weight COS (LMWCOS), and low molecular weight COS (LMWCOS)] according to their molecular weight, using an ultrafiltration membrane enzymatic bioreactor designed earlier . Antitumor activity of these COSs was then examined against Sarcoma 180 solid (S180) or Uterine cervix carcinoma No. 14 (Ul4) tumor cell-bearing mice. Among these COSs, MMWCOS with molecular weight range from 1.5 to 5.5 kDa effectively inhibited the growth of both tumor cells in the mice. In addition, the administration of MMWCOS resulted in increased thymus weight among lymphoid organs. The mice treated with MMWCOS showed improved survival rate and larger number of survivors after 40 days of feeding. The most effective of MMWCOS far antitumor activity in the S180- or U14-bearing mice was 20 mg/kg/day or more.
Identification and Phylogeny of the Human Endogenous Retrovirus HERV-W LTR Family in Human Brain cDNA Library and Xq21.3 Region
KIM, HEUI-SOO ; TIMOTHY J. CRO ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 508~513
Human endogenous retroviral long terminal repeats (LTRs) have been found to be coexpressed with sequences of genes located nearby. It has been suggested that the LTR elements have contributed to the structural change or genetic variation of human genome connected to various diseases. The HERV-W family has been identified in the cerebrospinal fluids and brains of individuals with schizophrenia. Using a cDNA library derived from a human brain, the HERV-W LTR elements were examined and five new LTR elements were identified. These elements were examined using a YAC clone panel from the Xq21.3 region linked to psychosis that was replicated on the Y chromosome after the separation of the chimpanzee and human lineages. Fourteen elements of the HERV-W LTR were identified in that region. Those LTR elements showed a high degree of sequence similarity (
) with previously reported HERV-W LTR. A phylogenetic tree obtained from the neighbor-joining method revealed that new HERV-W LTR elements were closely related to the AXt000960, AF072504, and AF072506 from the GenBank database. The data indicates that several copy numbers of the HERV-W LTR elements exist on the Xq21.3 region and are also expressed in the human brain. These LTR elements need to be further investigated as potential leads to neuropsychiatric diseases.
Detection of Oleic Acid Biodegradation by Fungi
Han, Dong-Wook ; Suh, Hwal ; Lee, Dong-Hee ; Park, Bong-Joo ; Kosuke Takatori ; Park, Jong-Chul ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 514~517
To investigate oleic acid biodegradation, 47 fungal strains were tested with modified Czapek Dox broth media containing oleic acid, and their biodegradative activities were assayed by measuring the release of
labeled oleic acid. After 72 h of cultivation, Aspergillus flavus, Aspergillus ochraceus, and Alternaria species metabolized approximately
of the supplied oleic acid. The relationship between the fungal degradation of oleic acid and the fungal growth was also examined using 7 strains of Aspergillus niger. A. niger. YMC 0100 and YMC 0322 degraded about
of the oleic acid after 72 h, while their germination ratios were more than
Cometabolic Production of Poly(3-Hydroxyalkanoates) Containing Carbon-Carbon Double and Triple Bonds by Pseudomonas oleovorans
Kim, Do-Young ; Kim, Young-Baek ; Rhee, Young-Ha ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 518~521
Poly(3-hydroxyalkanoate) copolyesters containing both carbon-carbon double and carbon-carbon triple bonds were produced by Pseudomonas oleovorans grown in mixtures of 10-undecynoic acid (10-UND(
)) and 10-undecenoic acid (10-UND(=)). The PHA content in the dry cells was usually 40 wt%. The bioconversion yield of (
) to PHA by P. oleovorans was remarkably enhanced from 1% to over 24% as the fraction of 10-UND(=) in the carbon substrate mixtures increased from 0 to 50%. These values were higher than those obtained when P. oleovorans was grown in the same molar mixtures of (
) and nonanoic acid (NA), indicating that 10-UND(=) was more efficient than NA as a cosubstrate in inducing cometabolic PHA production.
Rapid Separation of Cellular Cyclosophoraoses Produced by Rhizobium Species
Seo, Dong-Hyuk ; Lee, Sang-Hoo ; Park, Hey-Lin ; Kwon, Tae-Jong ; Jung, Seun-Ho ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 522~525
A very rapid and efficient separation technique for cellular rhizobial cyclosophoraoses was developed based on fractional precipitation and partition chromatography. Cyclosophoraoses are known to function in the osmotic regulation and root nodule formation of legumes during the nitrogen fixation process. Cyclosophoraoses are produced as unbranched cyclic (1longrightarrow12)-
-D-glucans in Agrobacterium or Rhizobium species. Recent research has shown that cyclosophoraoses can form inclusion complexation with various unstable or insoluble guest chemicals, thereby implying great potential for industrial application. Typical separation of pure cellular cyclosophoraoses has been so far carried out by several time-consuming steps, including size exclusion, anion exchange, and desalting liquid chromatographies, with a relatively poor recovery. However, the proposed method demonstrated that the successive application of fractional ethanol precipitation and one step of silica gel-based flash column chromatography was enough to simultaneously purify neutral or anionic forms of cyclosophoraoses. This novel technique is very rapid and provides a high recovery.
Hemolytic Activity of Culture Supernatant of Xenorhabdus nematophilus, a Symbiotic Bacterium of Entomopathogenic Nematodes
Ryu, Keun-Garp ; Bae, Jun-Sung ; Kwack, Kyu-Bum ; Kwon, O-Yul ; Park, Sun-Ho ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 526~529
Lysis of erythrocytes isolated from human, rabbit, and mouse blood samples was investigated with the culture supernatant of Xenorhabdus nematophilus in a primary form. Prior to use, the culture supernatant of the bacteria was concentrated and the concentrate was dialyzed against Tris-HCl buffer (10 mM, pH 8.1) by ultrafiltration using PM-5 membrane with a molecular weight cut-off of 5,000. At
, the supernatant exhibited no lytic activity towards three types of erythrocytes. However, at
, the supernatant showed selective lytic activity towards rabbit erythrocytes within 90 min. yet did not lyze human or mouse erythrocytes. Microscopic examination clearly revealed that most of the rabbit erythrocytes had been fumed into ghost forms.
Expression of Sortase, a Transpeptidase for Cell Wall Sorting Reaction, from Staphylococcus aureus ATCC 6538p in Escherichia coli
LEE, KI-YOUNG ; DONG-SUN SHIN ; JUNG-MIN YOON ; HEONJOONG KANG ; KI-BONG OH ;
Journal of Microbiology and Biotechnology, volume 12, issue 3, 2002, Pages 530~533
This paper describes the development of an enzymatic assay system for the identification of specific inhibitors of sortase, a transpeptidase that cleaves surface proteins of Cram-positive bacteria, from Staphylococcus aureus ATCC 6538p for antibacterial drug discovery. The coding region of the enzyme was amplified with the exception of the N-terminal membrane anchor sequence, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The enzyme activity was determined by quantifying increased fluorescence intensity upon cleavage of synthetic Dabcyl-QALPETGEE-Edans peptide. The results suggest that the developed in vitro assay system call be used in the search for sortase inhibitors In a short period of time.