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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
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Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 41, Issue 4 - Dec 2005
Volume 41, Issue 3 - Sep 2005
Volume 41, Issue 2 - Jun 2005
Volume 41, Issue 1 - Mar 2005
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Identification of the spk Gene Encoding Sphingosine Kinase in Sphingomonas chungbukensis DJ77 and Its Expression in Escherichia coli
Lee Su-Ri ; Um Hyun-Ju ; Kim Young-Chang ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 93~98
The sphingosine kinase gene, which is 969-nucleotide long, was identified during the whole genome sequencing of Sphingomonas chungbukensis DJ77. The amino acid sequence showed the identity of
with that of Zymomonas mobilis subsp. mobilis ZM4. C2, C3, and C5 domains of eukaryotic sphingosine kinase were found in sphingosine kinase from Sphingomonas chungbukensis DI77. One of these three conserved sites, GGDG, was predicted as a ATP-binding site, and the functions of the others were unknown currently. The phylogenetic tree constructed by ClustalX indicated that the sphingosine kinase of S. chungbukensis DJ77 was near the phylogenetic group COG1597, and did not belong to the group of diacylglycerol kinase of the same strain. The recombinant sphingosine kinase was expressed in Escherichia coli, but it was made in form of inclusion body.
Isolation and characterization of sigH from Corynebacterium glutamicum
Kim Tae-Hyun ; Kim Hyung-Joon ; Park Joon-Sung ; Kim Younhee ; Lee Heung-Shick ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 99~104
Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of
activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type
factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.
Construction and Analysis of a DNA Microarray for the Screening of Biosynthetic Genes of Secondary-Metabolites formation in Streptomyces
Nam Soo Jung ; Kang Dae-Kyung ; Rhee Ki Hyeong ; Kim Jong-Hee ; Kang Sang Sun ; Chang Yong Keun ; Hong Soon-Kwang ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 105~111
Streptomyces produces many kinds of secondary-metabolites including antibiotics. Screening of a new compound and elucidation of a biosynthetic pathway for the secondary metabolites are very important fields of biology, however, there is a main problem that most of the identified compounds are already researched compounds. To solve these problems, a microarray system that is based on the data related to the biosynthetic genes for secondary-metabolites was designed. For the main contents of DNA microarray, the important genes for the bio-synthesis of aminoglycosides, polyenes group, enediyne group, alpha-glucosidase inhibitors, glycopeptide group, and orthosomycin group were chosen. A DNA microarray with 69 genes that were involved in the bio-synthesis for the antibiotics mentioned above was prepared. The usability of the DNA microarray was confirmed with the chromosomal DNA and total RNA extracted from S. coelicolor whose genomic sequence had already been reported.
Salts Requirement of Moderately Halophilic Bacterium, Kordia algicida gen. nov., sp. nov.
Sohn Jae Hak ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 112~116
Moderately halophilic bacterium, Kordia algicida gen. nov., sp. novo was isolated from seawater of Masan Bay, Korea, during algal blooming caused by Skeletonema costatum. This bacterium was grown on the ZoBell 2216e medium supplied aged seawater, but not grown the medium supplied
NaCl. This bacterium showed absolute requirements for mono and divalent cations such as
, since no growth was observed in the medium, which is not supplemented with one of
ions. In kinetic studies for three kinds of cation, Km values of
were determined to 0.202 M, 0.089 mM, and 0.189 mM, respectively. Also,
was 0.442 h, 0.411 hand 0.316 h. The bacterial cells were quickly lysed in the condition limited by the cations. This result should be suggested that Kordia algicida originated from marine.
Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri
Moon Young-Gun ; Heo Moon-Soo ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 117~124
Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.
Nucleotide Sequence and Secondary Structure of 16S rRNA from Sphingomonas chungbukensis DJ77
Lee Kwan-Young ; Kwon Hae-Ryong ; Lee Won-Ho ; Kim Young-Chang ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 125~128
A 16S ribosomal RNA gene from S. chungbukensis DJ77 has been sequenced. This sequence had a length of 1,502 bp and was extended for 29 bp at 5' and for 37 bp at 3' from the partial sequence (1,435 bp) registered in 2000 year. Besides, 1 bp was newly added near to the 3' end. We made the secondary structure of the 16S rRNA based on E. coli model and found four specific regions. We found constant and variable regions in genus Sphingomonas as the result of multiple alignment of 16S rRNA gene sequences from Sphingomonas spp. and S. chungbukensis DJ77. We found a stem loop structure in S. chungbukensis DJ77, which was only discovered in C. jejuni to date. It showed the structural agreement despite the difference of the sequences from the both organisms. Finally, S. chungbukensis DJ77 belonged to cluster II (Sphingobium) group, after the classification using phylogenetic analysis and nucleotide signature analysis.
Isolation of Pigment Overproducing Mutant from Monascus purpureus and Optimization of Pigment Production
Park Chi Duck ; Jung Hyuck Jun ; Yu Tae Shick ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 130~134
Isolation of a pigment overproducing mutant, P-57, by ultraviolet irradiation of Monascus purpureus KCCM 60016 and investigation of the optimal conditions for pigment production of the mutant were carried out. P-57 mutant produced pigment on solid state culture. Unpolished rice was the best cereal source for pigment production among eight kinds of tested cereal sources for the solid culture of the mutant. The optimal culture condition for pigment production were obtained from the cultivated at
humidity for 30 days. The P-57 mutant strain showed the best pigment productivity of 160.0 unit at red pigment, 193.6 unit at orange pigment, and 141.6 unit at yellow pigment on solid state culture under optimal condition.
Antioxidant Activity of Monascus Pigment of Monascus purpureus P-57 Mutant
Park Chi Duck ; Jung Hyuck Jun ; Lee Hang Woo ; Kim Hyun Soo ; Yu Tae Shick ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 135~139
Antioxidant activity of monascus pigment of Monascus purpureus P-57 mutant was studied. Methanol extract from monascus pigment was separated into five organic solvent fractions; hexane, chloroform, ethyl acetate, butanol and water fractions. Hexane fraction showed the highest free radical scavenging effect on 1,l-diphenyl-2-picryl hydrazyl(DPPH), and the strongest inhibitory effect against xanthine oxidase, followed by chloroform fraction. But butanol and water fraction did not show inhibitory effect against the enzyme. Lineweaver-Burk plot showed that hexane fraction inhibited xanthine oxidase by non-competitive mode.
Transformation using Conjugal Transfer and attB Site Properties of Streptomyces natalensis ATCC27448
Lee Kang-Mu ; Choi Sun-Uk ; Park Hae-Ryong ; Hwang Yong-Il ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 140~145
Streptomyces natalensis ATCC27448 produces natamycin, a commercially important macrolide antifungal antibiotic. For molecular genetic study of S. natalensis, we have developed a system for introducing DNA into S. natalensis via conjugal transfer from Escherichia coli. An effective transformation procedure for S. natalensis was established based on transconjugation from E, coli ET12567/pUZ8002 using a
-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 10 mM
of E.coli donor cells without heat treatment of spores. In addition, southern blot analysis of exconjugants and the sequence of plasmids containing DNA flanking the insertion sites from the chromosome revealed that S. natalensis contains a single
attB site and at least a secondary or pseudo attB site. Similar to the case of various Streptomyces species, a single
attB site of S. natalensis is present within an ORF encoding a pirin-homolog, but a pseudo-attB site is present within a distinct site (GenBank accession no.
) and also its sequence deviates from the consensus sequences of attB sequence.
Expression of Aspergillus awamori Glucoamylase Gene in an Industrial Strain of Saccharomyces cerevisiae
Ghang Dong-Myeong ; Lee Su-A ; Chun Young-Hyun ; Chin Jong-Eon ; Lee Hwanghee Blaise ; Bai Suk ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 146~151
To construct an amylolytic industrial strain of Saccharomyces cerevisiae, the glucoamylase cDNA gene (GAl) from Aspergillus awamori was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) and integrated into the chromosomes of industrial S. cerevisiae. An integrative cassette lacking bacterial ampicillin resistance gene but containing the GA1 gene,
sequences of Ty1 retrotransposon as target sites for homologous recombination and S. cerevisiae aureobasidin A resistance gene (AUR1-C) as the selection marker was constructed to obtain a strain eligible for commercial use. Industrial S. cerevisiae transformed with this 15-integrative cassette efficiently secreted glucoamylase into the medium and grew on starch as the sole carbon source. The transformants were mitotically stable for 100 generations in nonselective medium.
Anti-Helicobacter pylori Activity of Pediococcus acidilactici GMB7330 Isolated from Infant Feces
Kang Ji-Hee ; Lee Myung-Suk ;
The Korean Journal of Microbiology, volume 41, issue 2, 2005, Pages 152~156
In the present study, lactic acid bacterium that has antibacterial activity against Helicobacter pylori was isolated from feces of newborn baby. The selection was based on the ability to inhibit the growth of H. pylori and to withstand harsh environmental conditions such as acidic pH and high bile concentration. By biochemical test and 16S rDNA sequencing, selected strain was turned out to be an Pediococcus acidilactici, therefore designated to P. acidilactici GMB7330. In order to investigate the inhibitory effects of P. acidilactici GMB7330 on the growth of H. pylori, we have tested in vitro studies such as cell viability and urease test. These results showed that antibacterial activity of P. acidilactici GMB7330 significantly decreased the viable cell count and urease activity of H. pylori. Antibacterial activity of P. acidilactici GMB7330 against H. pylori remained after pH adjustment to neutral, and the concentration of lactate produced from P. acidilactici GMB7330 was not enough to inhibit H. pylori. On the basis of the analysis by transmission electron microscope, it demonstrated that addition of P. acidilactici GMB7330 destroyed the cell structure of H. pylori. These results strongly suggested that P. acidilactici GMB7330 produce antibacterial substances to be able to inhibit the growth of H. pylori other than lactic acid.