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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
Journal Basic Information
Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 37, Issue 4 - Dec 2001
Volume 37, Issue 3 - Sep 2001
Volume 37, Issue 2 - Jun 2001
Volume 37, Issue 1 - Mar 2001
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Identification of Enteric Bacteria from Nephila clavata
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 1~8
Spiders are carnivores that prey upon insects and other small arthropods through digestion of food outside the body. Although spider poison may contain proteolytic enzymes, these are thought to play an insignificant role in actual digestion. The source of active proteolytic enzymes can be either the digestive tract cells of spider, or natural microbial flora in the digestive tract of spider. In this study, digestive tracts from the spider, Nephila clavata, were screened for bacteria that have protease or lipase activity. A total of
CFU was recovered from a spider and more than 90% of them showed protease and lipase activity respectively. Of the microbial isolates, 63.3% showed protease or lipase activity, and 50% of these showed both protease and lipase activity. Some of the isolates were characterized using a battery of chemical, phenotypic and genotypic methods. Eleven Gram negative bacteriaa (Acinetobacter calcoaceticus, A. haemolyticus, Alcaligenes faecalis, Cedecea davisae, C. neteri, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas fluorescens, Serratia marcescens, Stenotrophomonas maltophilia, Suttonella indologenes) and 11 Gram positive bacteria (Bacillus cereus, B. coagulans, B. pasteurii, B. thuringiensis, Cellulomonas flavigena, Corynebacterium martruchotii, Enterococcus durans, E. faecalis, Micrococcus luteus, Staphylococcus hominis, S. sciuri) were identified.
Molecular Taxonomy based on 16S rDNA Analysis and Pathogenicity of Yersinia pseudotuberculosis Isolated from Spring Waters
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 9~14
In order to investigate the pathogenicity and development of differential identification technique in the Yersinia species and other entericbacteria, we isolated 5 strains of Y.pseudotuberculosis from spring water sites in Seoul. The biochemical characteristics of isolated strains revealed that indole, VP(
, phenylalanine, lysine, arginine, ornithine, gas from glucose, lactose, sucrose, sorbitol, oxidase and motility(
) were all negative and urease, glucose, mannitol, salicin, catalase and motility(
) were all positive. To detect the causative agent of pseudotuberculosis(Y.pseudotuberculosis), we carried out a study using a PCR with inv primers complementary to the pathogenic region and found that all strains were positive, this revealed that strains from spring waters were pathogenic. Also 16S rDNA for total 5 strains of Y. pseudotuberculosis were amplified and a stretch of approximately 1,450 nucleotides were sequenced and analyzed. The 16S rDNA nucleotide sequence homologies among Yersinia species ranged 97.5% to 100% and between Y.pseudotuberculosis and other entericbacteria they ranged 93.0% to 95.1%. The Phylogenetic tree generated from the sequence analysis of the 16S rDNA gene showed 3 coherent clusters that could be separated into Y.pseudotuberculsis strains, some Yersinia species strains and other entericbacteria strains.
Characterization of the Bacterial Cell Wall Lytic Enzyme Produced by Aspergillus sp. HCLF-4
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 15~20
In this study, we have isolated bacterial cell wall lytic enzyme in the culture supernatant of Aspergillus sp. HCLF-4. This hydrolase showed cell wall lytic activity against Anabaena cylindrica. The extracellular enzyme was produced by Aspergillus sp. HCLF-4 when it was grown in a PDB media containing 0.05% heat killed Micrococcus luteus cells. The molecular weight of lytic enzyme was about 14.3 kDa. The optimal pH and temperature for the activity of this enzyme were 3.0~4.0 and
, respectively. This hydrolase activity was reduced by
, EDTA, and PMSF, whereas it was increased by
>. The enzyme has N-acetylmuramyl-L-amidase or endopeptidase activity.
Production of Bacillus anthracis Protective Antigen by Improvement of Culture Condition and Purification Methods
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 21~27
Recently many investigators have devoted considerable attention to the production and purification of PA for antigens, and the preparation of new synthetic medium (RM medium) have solved to increase the yields of the PA but, the low sensitivity of the PA to detect B. anthracis infections has remained as a problem to be solved. This study was undertaken to evaluate the yields of the PA from culture filtrates of B. anthracis Sterne
strain in modified RM medium in which 10 g/l of
and 10g/l of glucose were replaced by 8 g/l and 5 g/l, and the first purification step of PA from culture broth was used hydroxyapatite. The PA was purified by hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography and Toyo-pearl gel filtration chromatography. The yield of PA from the modified RM medium, 8.6 mg/l.
Characterization and Isolation of Mutants Involved in Cell Cycle Progression and Regulation in Saccharomyces cerevisiae
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 28~36
These studies were carried out to understand the mechanisms of genes which are related in cell cycle progression at G1/S phase. Mutants involved in cell cycle progression and regulation in Saccharomyces cerevisiae were isolated and characterized. To isolate new mutants, we screened the sensitivity to ciclopirox olamine (CPO) which inhibits the cell cycle traverse at or very near the G1/S phase boundary in HeLa cell and budding yeast. As results, we isolated 30 mutants and named cos(ciclopirox olamine sensitivity: cos27∼cos57) mutants. To determine the phenotype of mutants, we examined the sensitivity to methyl-methane sulfonate (MMS) and hydroxyurea (HU). Several mutants were sensitive to MMS and HU. According to these Phenotypes, cos mutants were grouped into four. Group I mutants are cos27, cos28, cos32, cos33, cos36, cos37, cos40, cos42, cos46, cos50, cos52 and cos53 which show MMS, HU sensitivities and might act at a checkpoint pathway during S phase. Group II mutants are cos43 and cos48 which show MMS sensitivities and might act at a checkpoint pathway during Gl or G2 phase. Group III mutants are cos35, cos47, cos54, cos55 and cos56 which show HU sensitivities and might act at a progress pathway during S phase. Finally, Group IV mutants are cos29, cos30, cos31, cos34, cos38, cos39, cos41, cos44, cos45, cos49, cos51 and cos57 which show only CPO sensitivities. Moreover, we examined the terminal phenotype of mutants under fluorescent microscope and then found one of S phase checkpoint related mutant(cos37). Furthermore, we constructed the heterozygote strain between mutant and wild type haploid strains to study their genetic analysis of cos mutants.
Nucleotide Sequence of 7.2 kb Mitochondrial Linear Plasmid DNA in Pleurotus ostreatus
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 37~41
Two linear plasmid-like DNAs, 10.2 kb and 7.2 kb were found in the mitochondria of P. ostreatus. They have covalently linked 5'-terminal proteins in both ends. Two continuous fragments of 4.7 kb and 2.3 kb from 7.2 kb DNA were cloned and sequenced. Two long open reading frames (ORF1; 2982 bp, 993 a.a and ORF2; 2703 bp, 900 a.a) and one short open reading frame(ORF3; 771 bp, 256 a.a) were found in the 7.2 kb plasmid. The putative ORF1 and ORF2 have conserved motifs of DNA polymerases and RNA polymerases, respectively, while the ORF3 has homologous regions with phosphatase from Plasmodium, and also with adhesine from Mycoplasma.
Isolation and Analysis of the Yeast Mutant Gene, soo1-1, which Confers the Defect in
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 42~48
Allele rescue and sequence analysis of soo1-1 allele in Saccharomyces cerevisiae mutant LP0353 revealed that soo1-1 is identical to the previously reported ret1-1 allele, which has a base substitution of A for
leading to an amino acid substitution of aspartic acid for
in Soolp. However, it was revealed that the addition of osmotic stabilizer, such as 1.2M sorbitol can rescue the temperature sensitive phenotype of the ret1-1 mutant and that the soo1-1/ret1-1 mutation may confer defects in post-translational modification of proteins involved in the yeast cell wall biogenesis. Evidence for a putative role of 5th WD40 domain of the Soo1p/
-COP in the construction and maintenance of cell walls was also presented by complementation test with deletion constructs of the SOOl.
Molecular Diversity of pagA Gene from Baciilus anthracis
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 49~55
Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. The anthrax toxin contains three components, including the protective antigen (PA), which binds to eucaryotic cell surface receptors and mediates the transport of toxins into the cell. In this study, the entire 2,294-nucleotide protective antigen gene (pag) was sequenced from 4 of B. anthracis strains to identify potential variation in the toxin and to further our understanding of B. anthracis evolution in Korea. Sequence alignment of the entire PA gene from 30 strains representative of the four B. anthracis diversity groups revealed mutations. The mutation of B. anthracis BAK are located adjacent to a highly antigenic region crossing the junction between PA domains 3 and 4 shown to be critical to LF binding. The different mutational combinations observed in this study give rise to 11 PA genotypes and 4PA phenotypes. Three-dimensional analysis of all the amino acid changes (Ala to Val) observed in BAK indicated that these changes are not only close sequentially but also very close in three-dimensional space to the antigenic region importan tfor LF binding. Phylogenetic (cladistic) analysis of the pag corresponded with previous strain grouping based on chromosomal variation, suggesting that plasmid evolution in B. anthracis has occurred with little or no horizontal transfer between the different strains.
Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 56~60
Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.
Improved Epifluorescence Microscopy for Observation of Phyllosphere Bacteria on Leaf Surfaces
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 61~65
Epifluorescence microscopy was used to observe epiphytic bacteria directly on plant leaf surfaces as well as indirectly in the leaf liberating solution by staining with fluorochromes of 4',6-diamidino-2-phenylindole (DAPI) and acridine orange(AO). Epiphytic bacteria could not be well observed on the leaf surface by staining with AO due to an intrusive orange or red background fluorescence. However, DAPI gave us clear epifluorescent images of the bacteria on the leaf. On the contrary, epiphytic bacteria in the liberating leaf solution were well observed on filters stained by both types of fluorochrome, although DAPI showed better fluorescent images than AO and not necessarily required a washing step of the filters stained. The optimum conditions of the DAPI stains were 5
g/ml for 5 min both for leaves and for filters of the liberating solution. It was confirmed that a critical step in the epifluorescence microscopy of leaf surfaces was to minimize release of water from the leaf. For this, the stained leaf samples were put on a filter paper, kept in a dry oven at
for 2 min instead of air-drying, and then immediately observed by epifluorescence microscopy. The established technique was applied to enumerate epiphytic bacteria on oak tree leaf surfaces.
Microbiological Elimination of 2,4-Dinitrotoluene and 276-Dinitro-toluene by an TNT-degrading Bacterium in Stirred Tank Reactors
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 66~71
An aerobic microbiological process was tested in 1.5 L stirred tank reactors for the treatment of dinitrotoluenes (DNTs)[e.g., 2.4-dinitrotoluene (2,4-DNT), 2,6-dinitrotoluene (2,6-DNT)] in the test culture of Stenotrophomonas maltophilia, which had previously characterized. Both 2,4-DNT and 2,6-DNT were completely degraded within 10 days and 14 days of incubation, respectively. Addition of the secondary carbon was essential to degrade DNTs, and little degradation was achieved in the absence of the secondary carbons. The effect of additional nitrogens on the degradation of DNTs was evaluated. Complete degradation of DNTs was observed in the absence of any additional nitrogens, whereas DNTs were partially degraded in the growth media with additional nitrogens. Both HPLC and GC-MS were used to detect and verify the residual DNTs and their intermediates. As the results, the HPLC and GC-MS chromatograms demonstrated that the both parent compounds, 2, 4-DNT and 2,6-DNT, and respective intermediates, 2-amino-4-nitrotoluene and 2-amino-6-nitrotoluene, could be successfully identified under the analytical conditions.
Characterization of Bacterial Community in the Ecosystem Amended with Phenol
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 72~79
The effect of phenol on the change of bacterial community in the effluent water from a wastewater treatment plant was analyzed by PCR and terminal restriction fragment length polymorphism (T-RFLP). The fragments of 16S rDNA were amplified by PCR with bacterial primers, where one of the primers was biotinylated at the 5'-end. After digestion with restriction enzymes, HaeIII and AluI, the biotinylated terminal restriction tragments (T-RFs) of the digested products were selectively isolated by using streptavidin paramagnetic particles. The single-stranded DNA of T-RFs was separated by electrophoresis on a polyacrylamide gel and detected by silver staining technique. When 10 standard strains were analyzed by our method, each strain had a unique T-RF which corresponded to the calculated size from the known sequences of RDP database. The T-RFLP fingerprint generated from the effluent water was very complex, and the predominant T-RFs corresponded to members of the genus Acinetobacter, Bacillus and Pseudomonas. In addition, the perturbation of bacterial community was observed when phenol was added to the sample at the final concentration of 250
. The number of T-RFs increased and the major bacterial population could be assigned to the genus Acinetobacter, Comamonas, Cytophaga and Pseudomonas. A intense band assigned to the putative genera of Acinetobacter and Cytophaga was eluted, amplified, and sequenced. The nucleotide sequence of the T-RF showed close relationship with the sequence of Acinetobacter junii.
The Seasonal Variation of Active Bacterial Abundance in Lake Soyang
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 80~84
Vertical and temporal variations of active and total bacterial abundance were monthly estimated in Lake Soyang from April 1999 to January 2000. The number of total and respiring bacteria was determined directly under microscope by AODC and CTC methods, respectively. The number of total and active bacteria varied from
, respectively. The proportions of respiring bacteria to total cell ranged from 3.7 to 44.2% : The proportions was the highest in November 1999 and the lowest in December 2000. The specific activity of
-glucosidase divided by total bacteria was
in August and
in September while the specific activity divided by CTC active bacteria was about
. The specific activity of active bacteria in September was 6.7 times higher than that of August. By these data of active bacteria, the new information of aquatic ecosystem was unveiled.
Microbial Community in Various Conditions of Soil Microcosm
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 85~91
Biological treatment of benzene and toluene contaminated soil was investigated in laboratory microcosm of 16 different types for degrading benzene and toluene by indigenous bacteria. At the experimental conditions of the microcosms fast degrading benzene and toluene, moisture contents were 30% and 60% in a soil gap and content of powdered-activated carbon(PCA) for adhesion of benzene and toluene-degrading bacteria was 1% in total soil mass. At the conclusion of the shifted bacteria community, Case 6 and case 7 were operated until 10 days, and then the total cell number and the number of benzene and toluene degrading bacteria were investigated. The total cell number of Case 6 and Case 7 increased 488 fold and 308 fold of total indigenous cell, respectively. The number of benzene and toluene degrading bacteria increased and maintained the percentages occupied in pre-operating microcosm. Species of benzene and toluene degrading bacteria in microcosm changed from species of Gram negative bacteria to Gram positive bacterial species after soil exposed to benzene and toluene.
Economic Consideration of Poly(3-hydroxybutyrate) Production by Fed-batch Culture of Ralstonia eutropha KHB 8862
The Korean Journal of Microbiology, volume 37, issue 1, 2001, Pages 92~99
High-cell-density cultivation of Ralstonia eutopha KHB 8862 by fed-batch fermentation in a 200 l pilot plant was carried out for the mass production of poly(3-hydroxybutyrate) (PHB). After 80 h of cultivation, the dry cell weight (DCW), PHB concentration, and PHB yield from fructose syrup reached 168 g/l, 74%DCW, and 0.27 (w/w), respectively, resulting in a productivity of 1.6 g of PHB/L/h. Based on these results, the PHB production cost from bacterial fermentation was analyzed and economic evaluation was performed. In the case of new investment being implemented or not, the production cost of PHB was US$ 3.15/kg and US$ 2.41/kg, respectively. PHB productivity and PHB yield on a carbon substrate were both important factors to be optimized. The increase of PHB yield on a carbon sources significantly decreased the PHB production cost but the increase in productivity had a relatively slight effect on the decrease in PHB production cost because the cost of carbon sources (37%) for PHB was larger in proportion to total cost than the depreciation cost (17%). These results suggest that the increased PHB yield from carbon sources and the development of new cheaper substrates would be more effective in decreasing PHB production cost than the increase in productivity. It was demonstrated that PHB is not in competition with consumable plastics such as PET in present market. Therefore, it is essential to lower production cost to be used as a bulk product and desirable to develop new application fields for PHB such as biomedical and cosmeceuticals.