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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
The Korean Journal of Microbiology
Journal Basic Information
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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Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1
Lim Hyun Sook ; Goo Jae Whan ; Kim Lee Hyun ; Kim Si Wouk ; Cho Eun Hee ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 1~7
Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.
A spsB Gene Putatively Encoding Glucosyl-Isopreny Phosphate-Transferase in Sphingomonas chungbukensis DJ77
Lee Soo-Youn ; Choi Jung-Do ; Shin Malshick ; Kim Young-Chang ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 8~12
Some genes, which are involved in the biosynthesis of polysaccharides, could be found by the genome project of Sphingomonas chungbukensis DJ77. In this study, we identified the complete nucleotide sequence of a gene, encoding the glucosyl-isoprenyl phosphate-transferase, which catalyzes the first step in the biochemical pathway for the synthesis of the sphingan type polysaccharide. This gene, named spsB, is initiated by the ATG codon and terminated by the TGA, and its open reading frame consists of 1392 bp, encoding 463 amino acids. The predicted amino acid sequence of this enzyme indicates
similarity to SpsB of Sphingomonas spp S88, also produces sphingan, and
to GelB of Sphingomonas paucimobilis ATCC 31461.
Analysis of N- Terminal Amino Acid Sequence of Catechol 2,3-dioxygenase from Aniline Degrading Delftia sp. JK-2
Hwang Seon-Young ; Kahng Hyung-Yeel ; Oh Kye-Heon ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 13~17
The aim of this work was to investigate the N-terminal amino acid sequence of catechol 2,3-dioxygenase isolated from Delftia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O from strain JK-2 was
, and exhibited high sequence similarity with that of C2,3O from Pseudomonas sp., Comamonas sp. JS765, Comamonas test-osteroni, or Burkholderia sp. RP007. Approximately 950-bp C2,3O was obtained through PCR using the primers derived from N-terminal amino acid sequence. Analysis of the DNA sequence revealed that the deduced 296 amino acid sequences were determined, and it showed
identity with C2,3O from Pseudomonas sp. AW-2 and
similarity with Comamonas sp. JS765.
Physiological Characterization of Lactobacillus sp. JK-8 Isolated from Shrimp Aquaculture Pond
Chun Jae-Woo ; Ma Chae-Woo ; Oh Kye-Heon ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 18~23
The purpose of this work was to investigate the physiological characteristics of Lactobacillus sp. JK-8 isolated from a shrimp aquaculture pond. The strain JK-8 was grown on MRS media, and morphological and physiological characteristics of the strain were examined. The bacterium was identified as a strain of the genus Lactobacillus on the basis of BIOLOG test. Strain JK-8 produced both lactic acid and acetic acid, which were responsible for the pH decrease during growth. Concentrations of lactic acid and acetic acid increased to 192.8 mM and 43.6 mM, respectively, and the initial pH 7.0 of the cultures decreased to 3.8 at the end of incubaction. The bacteriocidal effect against eight target bacteria was examined with 5-fold concentrated culture supernatants. All bacteria tested in this work were completely killed within 3 hrs after treatment with the culture supernatant. The bacteriocidal effects were clearly observed, only when the pH of the culture supernatants were not adjusted. HPLC was used to reslove lactic acid and acetic acid in the culture solution, and GC-MS was used to verify the metabolites.
Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR
Woo Yong-Ku ; Kim Shin ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 24~37
This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains
tested and followed by coa-gene
, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains
were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.
Microbial Hygienic Status of Poultry Meats and Eggs Collected at the Public Markets in Seoul and Kyung-gi Regions in 1996
Woo Yong-Ku ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 38~46
To determine the actual hygienic status of domestic chicken meats sold in public markets (conventional markets and department stores), microbial contamination levels (Total cells, Coliforms and Staphylococcal cells) and zoonotic pathogens (Salmonella species, Campylobacter species, Listeria species, and Staphylococcus aureus) isolation tests were conducted. Chicken meats and eggs tested were collected from the conventional markets (Si-Jang) and department-stores located in Seoul and Kyung-gi regions in 1996. In total cells and coliforms contamination tests, chicken meats sold in department stores were much lesser contamination status than those of Si-Jang, but staphylococcal cells level was much more higher than that of conventional markets. Salmonella isolation frequency was investigated as
, but Campylobacter jejuni and Listeria monocytogenes isolation frequency were appeared both
. In case of eggs sold in public markets, one of S. gallinarum strain
was isolated only on the egg-shell part among the four-hundred and fourty-six. In comparison with foreign imported chicken meats, there were no big differences in microbial contamination status. On the other hand, both Salmonella and L. monocytogenes were isolated only in the chicken wings from Korea and China, but not from U.S.A. This data suggest that more hygienic control system in order to produce the safe and hygienic chicken meats and eggs is need in our country as soon as possible.
Identification of Bacterial Strains Adhered to Human Scalp Hair and Antimicrobial Susceptibility
Lee Moon Sook ; Han Hyo Shim ; Jung Jae Sung ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 47~52
This study was carried out to identify bacterial strains adhered to human scalp hair and to investigate the antibiotic susceptibility of them. A total of 39 isolates were obtained from patients in intensive care units and healthy persons. The most common species isolated was Staphylococcus epidermidis (19 isolates), followed by S. aureus (14 isolates), S. waneri (5 isolates), and S. pasteuri (1 isolate). The susceptibility of isolates to amikacin, ampicillin, bacitracin, carbenicillin, cefazolin, cefoperazone, chloramphenicol, erythromycin, gentamicin, methicillin, nalidixic acid, neomycin, oxacillin, penicillin, streptomycin, tetracycline and vancomycin was determined by the disk diffusion method. All of the antibiotic resistant isolates were obtained from patient scalp hair. To examine the effect of conventional shampoo and detergent SDS on removing of bacteria from hair, we treated hair with culture solution of S. aureus. The bacteria attached to hair were not removed even by repeated washing with detergents. These results suggested that hair could be a source of bacterial contamination in hospital.
Analysis of Influenza Virus Isolates in Seoul during 2003-2004 Season
Hwang Young-Ok ; Lee Jae-In ; Seo Byung-tae ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 53~59
Influenza is an important public health problem which occurs almost every winter in temperate climates and is often associated with increased rates of hospitalization and death. In 1999, our influenza surveillance was initiated with 4 voluntary sentinel physicians and the Public Health Center. During the 2003-2004 influenza season, 124 influenza viruses were isolated from 401 clinical specimens, which were collected from patients with Influenza-like illness(ILI) in Seoul. The case definition of ILI is a case with fever more than
and systemic symptoms; cough, or sore throat. ILI was the highest at the 20-49 age
and the rate of virus isolation was the highest at the 7-19 age
. Among 124 influenza viruses, isolates 83 were identified as A/H3N2 type and others were subtyped as influenza B viruses in 2003-2004 season. Influenza viruses were collected
Gangnam-Gu and Seocho-Gu etc. and the isolate rate of virus had the area difference; Yongsan-Gu
. Out of 401 individuals, 160 was vaccinated
and the vaccination rate was the highest at the 20-49 age
. These findings may contribute to the recommondation of the influenza vaccine formulation and the development of influenza control measure.
Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis
Woo Yong-Ku ; Kim Shin ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 60~66
To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.
Enhanced PHB Accumulation in Photosystem- and Respiration-defective Mutants of a Cyanobacterium Synechocystis sp. PCC 6803
Kim Soo-Youn ; Choi Gang Guk ; Park Youn Il ; Park Young Mok ; Yang Young Ki ; Rhee Young Ha ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 67~73
Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to
of dry cell weight (DCW) comparable to
of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of
ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.
Antioxidant Activity and Characterization of Exiguobacterium sp. SC2-1 Isolated from Sea Water
Kim Man-Chul ; Park Guen- Tae ; Son Hong-Joo ; Choi WooBong ; Heo Moon-Soo ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 74~80
For the reseach of the natural marine antioxidant, an antioxidant-producing bacterium was isolated from seawater in Jeju costal area. The isolated strain SC2-1 was Gram-positive, catalase positive, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and NaCl required for growth. The radical scavenging activity of the culture supernatant was detemined by DPPH method. This bacterium was identified based on morphological, biochemical characteristics and 16S rDNA sequence analysis, and then was named Exiguobacterium sp. SC2-1. The optimum conditions of culture for production antioxidnat were
, pH 6-8 and
NaCl. The stain showed the highest activity and growth cultured in medium which added
maltose. Hydroxyl radical activity of the supernatant of Exiguobacterium sp. SC2-1 was
. The SOD activity of the culture supernatant was estimated about
Characterization of Endoglucanase (F-I-III) Purified from Trichoderma sp. C-4
Sul Ok Ju ; Chung Dae Kyun ; Han In Seob ; Jeong Choon Soo ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 81~86
One of the endoglucanases, F-I-III, was purified from the culture filtrate of T. sp. C-4 through procedures including chromatography on Sephacryl S-200, DEAE-Sepharose A-50, and Chromatofocusing on Mono-P (FPLC). The molecular weight of the enzyme was determined to be about 56,000 Da by SDS-PAGE, and pI of 4.9 by analytical isoelectric focusing. F-I-III showed the highest enzyme activity at
, and the pH optimum of the enzyme was 5.0. There was no loss of activity when the enzyme was incubated at
for 24 hours. The specific activity of the enzyme F-I-III toward the CMC was 315.4 U/mg. The Km value for
of F-I-III was 2.69 mM. N-terminal sequence of F-I-III was analyzed to be QPGTSTPEVHPKKLTTYK. It showed
of homology to that of EGI from T. reesei. The presence of some metal ions (1 mM) had only a little effect on CMCase activity. The treatment of the reducing agents resulted in the increase of endoglucanase activity.
Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production
Hwang Ji-Hwan ; Lee Chang-Kwon ; Lee Kang-Mu ; Jo Byoung-Kee ; Park Hae-Ryong ; Hwang Yong-Il ;
The Korean Journal of Microbiology, volume 41, issue 1, 2005, Pages 87~92
Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.