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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 16, Issue 12 - Dec 2006
Volume 16, Issue 11 - Nov 2006
Volume 16, Issue 10 - Oct 2006
Volume 16, Issue 9 - Sep 2006
Volume 16, Issue 8 - Aug 2006
Volume 16, Issue 7 - Jul 2006
Volume 16, Issue 6 - Jun 2006
Volume 16, Issue 5 - May 2006
Volume 16, Issue 4 - Apr 2006
Volume 16, Issue 3 - Mar 2006
Volume 16, Issue 2 - Feb 2006
Volume 16, Issue 1 - Jan 2006
Selecting the target year
A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line
Demir, Ismail ; Demirbag, Zihni ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1485~1490
In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose (
) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as
plaque forming units (PFU)/ml and
, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.
Intratumoral Administration of Rhenium-188-Labeled Pullulan Acetate Nanoparticles (PAN) in Mice Bearing CT-26 Cancer Cells for Suppression of Tumor Growth
Song, Ho-Chun ; Na, Kun ; Park, Keun-Hong ; Shin, Chan-Ho ; Bom, Hee-Seung ; Kang, Dong-Min ; Kim, Sung-Won ; Lee, Eun-Seong ; Lee, Don-Haeng ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1491~1498
The feasibility of pullulan acetate nanoparticles (PAN) with ionic strength (IS) sensitivity as a radioisotope carrier to inhibit tumor growth is demonstrated. PAN was radiolabeled with rhenium 188 (Re-188) without any chelating agents. The labeling efficiency of Re-188 into PAN (Re-188PAN) was
as determined by TLC. The tumor volumes of mice treated with 0.45 mCi of Re-188-PAN were measured and compared with that of free Re-188 after 5 days of intratumoral injection. For the histological evaluation of apoptotic nuclei of tumor cells, hematoxylin and eosin (H&E), and terminal deoxynucleotidyl transferase biotinylated deoxyuridine triphosphate nick end labeling (TUNEL) staining were performed. The mean tumor volume of the Re-188-PAN-treated group was decreased by 36% after 5 days, whereas that the free Re-188-treated group was decreased by only 15% (P<0.05). The mean number of TUNEL-positive cells in Re-188-PAN-treated tumors at
cells/section was significantly greater than the control (
cells/section, P=0.03). The numbers of leukocyte and lymphocyte were decreased in both free Re-188- and Re-188-PAN-treated mice. These results indicated that the intratumoral injection of Re-188-PAN effectively inhibits the tumor growth by prolonging Re-188 retention time in tumor site induced by the IS sensitivity.
Two-Dimensional Reference Map of Schizosaccharomyces pombe Proteins (Update)
Kim, Sun-Kyung ; Won, Mi-Sun ; Sun, Nam-Kyu ; Jang, Jae-Won ; Lee, Seung-Hee ; Shin, Hee-Young ; Song, Kyung-Bin ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1499~1512
Based on the first 2D reference map of the fission yeast Schizosaccharomyces pombe protein reported previously, we expanded and updated the map using narrower pI ranges. In this paper, 240 protein spots were identified on our reference map. In the pI 4-7 range, 144 spots corresponding to 86 different proteins were identified. In the pI 6-9 range, 43 spots corresponding to 35 different proteins were identified. Fifty-three new spots corresponding to 39 different proteins were further identified in the pI 5-6 range.
Isolation of Lactobacillus plantarum from Kimchi and Its Inhibitory Activity on the Adherence and Growth of Helicobacter pylori
Lee, Hak-Mee ; Lee, Yeon-Hee ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1513~1517
One single lactic acid producing bacterium, isolated from kimchi, inhibited the growth and adherence of Helicobacter pylori to the human gastric epithelial cell line MKN-45. This isolate was identified as Lactobacillus plantarum and termed L. plantarum strain PL9011. The adherence of H pylori, in the presence of live or nonviable L. plantarum strain PL9011 (10-fold CFU), decreased to 14-20%. The spent culture supernatant of L. plantarum strain PL9011 resulted in the eradication of H pylori. This activity remained stable following neutralization and heat treatment, but not following pepsin treatment, thereby suggesting small peptides as the inhibitory factor. L. plantarum strain PL9011 did not produce any harmful metabolites or enzymes. The results obtained in this study suggest that the L. plantarum strain PL9011 may be a potential novel probiotic for the stomach.
Dynamics Behavior of Phage-Host System Related to Microlunatus phosphovorus in Activated Sludge with Host Inoculation
Lee, Sang-Hyon ; Otawa, Kenichi ; Onuki, Motoharu ; Satoh, Hiroyasu ; Mino, Takashi ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1518~1522
In the present study, it was observed how the phage-host system that is naturally reproduced in activated sludge is affected by the host inoculation. The system of Microlunatus phosphovorus and its phages was selected as the phage-host system native to an activated sludge system operated for 19 days under sequencing anaerobic-aerobic conditions with glutamate as the main carbon source. The phage-host system related to M. phosphovorus was monitored by plaque assay for the phages and by fluorescent in situ hybridization (FISH) for the bacterial host. In addition, the whole phage structure was also monitored by pulsed-field gel electrophoresis (PFGE). During the first 9 days, the phage-host system was more or less steady at approx. 9% (FISH/ DAPI) for M. phosphovorus and approx. 10,000 PFU/ml for its lytic phages. Microlunatus phosphovorus JCM9379 was inoculated into the activated sludge on day 10. Right after the inoculation, M. phosphovorus was approx. 24% (FISH/DAPI) whereas its lytic phages dropped down to approx. 500 PFU/ ml. After the host inoculation (within 9 days), however, the phage-host system eventually reverted to its original level in each population. On the other hand, the whole phage structure was not significantly changed by M. phosphovorus inoculation but stable throughout the process operation. Only the minor change that four phage groups gradually became abundant after the host inoculation was observed.
Optimization of Quartz Crystal Microbalance-Precipitation Sensor Measuring Acetylcholinesterase Activity
Kim, Nam-Soo ; Park, In-Seon ; Kim, Dong-Kyung ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1523~1528
The optimization of a batch-type quartz crystal microbalance (QCM)-precipitation sensor measuring acetylcholinesterase (AChE) activity was conducted. To covalently bind AChE onto the gold electrode of a QCM surface, glutaraldehyde cross-linking to a cystamine self-assembled monolayer was tried at different cystamine concentrations. At the optimum conditions of the QCM-precipitation sensor, 0.1 M potassium phosphate buffer (pH 8.0), containing 0.01% Tween 80, was used as the reaction buffer, with the enzyme amount of 5 units for immobilization and the substrate concentration of 50 mg/ml. The current biosensor might find a future applicability to the sum parameter detection on organophosphorus and carbamate pesticides.
Immunological Characterization of Full and Truncated Recombinant Clones of ompH(D:4) Obtained from Pasteurella multocida (D:4) in Korea
Kim, Young-Hwan ; Cheong, Ki-Young ; Shin, Woo-Seok ; Hong, Sung-Youl ; Woo, Hee-Jong ; Kwon, Moo-Sik ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1529~1536
We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea . The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their
were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some
of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at
of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose (
) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.
Polyphasic Assignment of a Highly Proteolytic Bacterium Isolated from a Spider to Serratia proteamaculans
Kwak, Jang-Yul ; Lee, Dong-Hun ; Park, Youn-Dong ; Kim, Seung-Bum ; Maeng, Jin-Soo ; Oh, Hyun-Woo ; Park, Ho-Yong ; Bae, Kyung-Sook ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1537~1543
A bacterial strain named HY-3 that produces a highly active extracellular protease was isolated from the digestive tract of a spider, Nephila clavata. The bacterium was a Gram-negative, oxidase-negative, catalase-positive, nonhalophilic, nitrate-reducing, facultative anaerobe. Transmission and scanning electron microscopies demonstrated that the isolate was non-spare-forming, straight, rod-shaped, and motile by peritrichous flagella. The G+C content of the DNA was 57.0 mol%. The isoprenoid quinone type was ubiquinone with 8 isoprene units (Q-8). The morphological and biochemical characteristics including the predominant fatty acid and phospholipids profiles placed the isolate HY-3 in the family Enterobacteriaceae. Further biochemical characterization and phylogenetic studies including determination of an almost complete 16S ribosomal DNA sequence suggested that the bacterium was closely related to the genus Serratia. DNA-DNA hybridization analysis revealed that this extracellular protease-producing strain belongs to Serratia proteamaculans, which is also known far its association with insects.
Identification of Anti-Angiogenic and Anti-Cell Adhesion Materials from Halophilic Enterobacteria of the Trachurus japonicus
Lim, Jong-Kwon ; Seo, Hyo-Jin ; Kim, Eun-Ok ; Meydani, Mohsen ; Kim, Jong-Deog ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1544~1553
The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of
) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited cell adhesion that non-activated U937 monocytic cell adhesion to HUVECs activated with
by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with
to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to
-stimulated HUVECs was completely suppressed by 121% at a concentration of
. PLM-f74 blocked signal pathways from VEGFR2, PI3K,
-catenin, and VE-cadherin to NF-kB, based on western bolt analysis. It also inhibited IL-l-stimulated HUVEC expression of the adhesion molecules, ICAM-l by 40%, VCAM-l by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities, and non-toxicity may be expected from these results.
Bacillus ginsengihumi sp. nov., a Novel Species Isolated from Soil of a Ginseng Field in Pocheon Province, South Korea
Ten Leonid N. ; Im Wan-Taek ; Baek Sang-Hoon ; Lee, Jung-Sook ; Oh, Hee-Mock ; Lee, Sung-Taik ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1554~1560
A Gram-positive, aerobic or facultative anaerobic, non motile, endospore-forming bacterial strain, designated Gsoil
, was isolated from a soil sample of a ginseng field in Pocheon Province (South Korea), and was characterized taxonomically by using a polyphasic approach. It grew well on nutrient agar medium and utilized a limited number of organic substrates as sole carbon sources, including D-xylose and some other carbohydrates, but did not utilize L-amino acids and organic acids. The isolate was positive for oxidase test but negative for catalase, and negative for degradation of macromolecules such as starch, cellulose, xylan, casein, chitin, and DNA. The G+C content of the genomic DNA was 41.8 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were
(30.2%). Comparative 16S rRNA gene sequence analysis showed that strain Gsoil
fell within the radiation of the cluster comprising Bacillus species and joined Bacillus shackletonii LMG
with a bootstrap value of 95%. The highest 16S rRNA gene sequence similarities were found with Bacillus shackletonii LMG
(97.6%), Bacillus acidicola DSM
(96.9%), Bacillus sporothermodurans DSM
(96.5%), and Bacillus oleronius DSM
(96.5%). The phylogenetic distance from any other validly described species within the genus Bacillus was less than 96%. DNA-DNA hybridization experiments showed that the DNA-similarities between strain Gsoil
and closest phylogenetic neighbors were less than 39%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil
) was classified in the genus Bacillus as the type strain of a novel species, for which the name Bacillus ginsengihumi sp. nov. is proposed.
Improvement of PCR Amplification Bias for Community Structure Analysis of Soil Bacteria by Denaturing Gradient Gel Electrophoresis
Ahn, Jae-Hyung ; Kim, Min-Cheol ; Shin, Hye-Chul ; Choi, Min-Kyeong ; Yoon, Sang-Seek ; Kim, Tae-Sung ; Song, Hong-Gyu ; Lee, Geon-Hyoung ; Ka, Jong-Ok ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1561~1569
Denaturing gradient gel electrophoresis (DGGE) is one of the most frequently used methods for analysis of soil microbial community structure. Unbiased PCR amplification of target DNA templates is crucial for efficient detection of multiple microbial populations mixed in soil. In this study, DGGE profiles were compared using different pairs of primers targeting different hypervariable regions of thirteen representative soil bacteria and clones. The primer set (1070f-1392r) for the E. coli numbering 1,071-1,391 region could not resolve all the 16S rDNA fragments of the representative bacteria and clones, and moreover, yielded spurious bands in DGGE profiles. For the E. coli numbering 353-514 region, various forward primers were designed to investigate the efficiency of PCR amplification. A degenerate forward primer (F357IW) often yielded multiple bands for a certain single 16S rDNA fragment in DGGE analysis, whereas nondegenerate primers (338f, F338T2, F338I2) differentially amplified each of the fragments in the mixture according to the position and the number of primer-template mismatches. A forward primer (F352T) designed to have one internal mismatch commonly with all the thirteen 16S rDNA fragments efficiently produced and separated all the target DNA bands with similar intensities in the DGGE profiles. This primer set F352T-519r consistently yielded the best DGGE banding profiles when tested with various soil samples. Touchdown PCR intensified the uneven amplification, and lowering the annealing temperature had no significant effect on the DGGE profiles. These results showed that PCR amplification bias could be much improved by properly designing primers for use in fingerprinting soil bacterial communities with the DGGE technique.
Genetic Characterization of Encephalomyocarditis Virus Isolated from Aborted Swine Fetus in Korea
Song, Min-Suk ; Joo, Young-Ho ; Lee, Eun-Ho ; Shin, Jin-Young ; Kim, Chul-Jung ; Shin, Kwang-Soon ; Sung, Moon-Hee ; Choi, Young-Ki ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1570~1576
An encephalomyocarditis virus (EMCV-CBNU) was isolated from an aborted swine fetus in October 2005. To investigate the genetic origin and virulence of the EMCV-CBNU strain, we determined the complete sequence of the virus and tested its virulence in mice. Genetic characterization revealed that the RNA genome was composed of 7,713 nucleotides with a single open reading frame (2,292 amino acids), coding 12 proteins. The EMCV-CBNU had the shortest poly(C) tract, consisting of 10 C's (
), compared with all the other EMCV strains reported in GenBank. Amino acid and phylogenetic analyses showed that EMCV-CBNU had the highest genetic identity with strain 2887A (99.7%), which was originally isolated from a fetus in a pig breeding farm that had a history of reproductive failure. Because rodents are the natural host of EMCV, we investigated the virulence of EMCV-CBNU in mice. Surprisingly, all mice inoculated with more than
of EMCV-CBNU showed symptoms of hind limb paralysis and eventually died during 3 and 8 days postinoculation (DPI). Furthermore, when we inoculated the virus into pregnant mice, all dams and their fetuses died in 6 DPI. This is the first report on a full genomic analysis of swine EMCV in Korea, which exhibits high virulence in mice.
Regeneration of a Cartilage Tissue by In Vitro Culture of Chondrocytes on PLGA Microspheres
Son, Jeong-Hwa ; Park, So-Ra ; Kim, Hyeon-Joo ; Min, Byoung-Hyun ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1577~1582
Cartilage tissue engineering has emerged as an alternative approach for reconstruction or repair of injured cartilage tissues. In this study, rabbit chondrocytes were cultured in a three-dimensional environment to fabricate a new cartilaginous tissue with the application of tissue engineering strategies based on biodegradable PLGA microspheres. Chondrocytes were seeded on PLGA microspheres and cultured on a rocking platform for 5 weeks. The PLGA microspheres provided more surface area to adhere chondrocytes compared with PLGA sponge scaffolds. The novel system facilitated uniform distribution of the cells on the whole of the PLGA microspheres, thus forming a new cartilaginous construct at 4 weeks of culture. The histological and immunohistochemical analyses verified that the number of chondrocytes and the amount of extracellular matrix components such as proteoglycans and type II collagen were significantly greater on the PLGA microspheres constructs as compared with those on the PLGA sponge scaffolds. Therefore, PLGA microspheres enhanced the function of chondrocytes compared with PLGA sponge scaffolds, and thus might be useful for formation of cartilage tissue in vitro.
High-Level Expression of T4 Endonuclease V in Insect Cells as Biologically Active Form
Kang, Chang-Soo ; Son, Seung-Yeol ; Bang, In-Seok ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1583~1590
T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.
Color Alteration and Acaricidal Activity of Juglone Isolated from Caesalpinia sappan Heartwoods Against Dermatophagoides spp.
Lee, Chi-Hoon ; Lee, Hoi-Seon ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1591~1596
Acaricidal effects of materials derived from Caesalpinia sappan heartwoods against Dermatophagoides farinae and D. pteronyssinus were assessed and compared with those evidenced by commercial benzyl benzoate and DEET. The observed responses varied according to dosage and mite species. The
values of the methanol extracts derived from C. sappan heartwoods were 6.13 and
against D. farinae and D. pteronyssinus, respectively. Furthermore, the ethyl acetate fraction derived from the methanol extract was approximately 8.71 more toxic than DEET against D. farinae, and 4.73 times more toxic against D. pteronyssinus. The biologically active constituent from the ethyl acetate fraction of C. sappan heartwood extract was purified via silica gel chromatography and high-performance liquid chromatography. The structure of the acaricidal component was analyzed by
, and DEPT-NMR spectroscopy, and identified as juglone (5-hydroxy-l,4-naphthoquinone). Based on the
values of juglone and its derivatives, the most toxic compound against D. farinae was juglone (
), followed by benzyl benzoate (
) and 2methyl-l,4-naphthoquinone (
). These results indicate that the acaricidal activity of C. sappan heartwoods is likely to be the result of the effects of juglone. Additionally, juglone treatment was shown to effect a change in the color of the cuticles of house dust mites, from colorless-transparent to dark brownish-black. Accordingly, as a naturally occurring acaricidal agent, C. sappan heartwood-derived juglone should prove to be quite 'useful as a potential control agent, lead compound, and house dust mite indicator.
Rahnella aquatilis Strain AY2000 Produces an Anti-Yeast Substance
Ryu, Eun-Ju ; Kim, Han-Woo ; Kim, Byung-Woo ; Kwon, Hyun-Ju ; Kim, Kwang-Hyeon ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1597~1604
To screen for an anti-yeast substance (AYS), many bacteria were isolated from soil and a strain AY2000 was selected. The strain AY2000 was identified as Rahnella aquatilis by morphology, biochemical properties, and 16S r-RNA nucleotide sequence analyses. The strain AY2000 showed anti-yeast activity against Candida albicans and Saccharomyces cerevisiae, whereas R. aquatilis ATCC33071 as a type strain did not show the activity against the yeasts under the same condition. The growth of yeast cell was significantly inhibited by AYS produced by the strain AY2000, as shown by optical density and MTT assay. The minimum inhibitory concentration (MIC) of the AYS against S. cerevisiae and C. albicans at
, respectively. The MIC of AYS against hyphae of C. albicans at
. Scanning electron microscopic analysis revealed that yeast cells treated with AYS had an irregular form with a wrinkled and rough surface.
Synergistic Effect of Interleukin-18 on the Expression of Lipopolysaccharide-Induced IP-10 (CXCL-10) mRNA in Mouse Peritoneal Macrophages
Kim, Hyo-Young ; Kim, Jae-Ryong ; Kim, Hee-Sun ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1605~1612
Interleukin (IL)-18, a member of the family of IL-l cytokine, is one of the principal inducers of
in T lymphocytes and natural killer cells. The objective of the present study was to evaluate the effect of IL-18 on the expression of chemokine IP-10 (CXCL-10) mRNA in mouse peritoneal macrophages. IL-18 had very weak direct effect or synergistic effect with IL-12 on the expression of IP-10 mRNA in C57BL/6 mouse peritoneal macrophages. However, IL-18 pretreatment was found to playa cooperative role in the expression of lipopolysaccharide (LPS)-induced IP-10 mRNA. For the expression of LPS-induced IP-10 mRNA, the synergistic effect was detected after 16 h of IL-18 pretreatment prior to LPS stimulation. The expression level of CD14 in cells stimulated with LPS was not changed by IL-18 pretreatment, and the level of
production during IL-18 pretreatment plus LPS stimulation was barely discernible (
). Namely, the synergistic effect of IL-18 pretreatment was not related to a change of LPS receptor, CD14 expression, and the production of
by the interaction between IL-18 and LPS. The synergistic effect of IL-18 pretreatment on the expression of LPS-induced IP-10 was related to not NF-kB but AP-1 activation, and associated with the extracellular signal-regulated kinase (ERK) pathway, one of the mitogen-activated protein kinase signaling pathways. These results provide useful information that may elucidate the mechanisms underlying the effect of IL-18 on the expression of IP-10 mRNA.
Antialgal Effect of a Novel Polysaccharolytic Sinorhizobium kostiense AFK-13 on Anabaena flos-aquae Causing Water Bloom
Kim, Jeong-Dong ; Lee, Choul-Gyun ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1613~1621
Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of
-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.
Plant Growth Substances Produced by Methylobacterium spp. and Their Effect on Tomato (Lycopersicon esculentum L.) and Red Pepper (Capsicum annuum L.) Growth
Ryu, Jeong-Hyun ; Madhaiyan, Munusamy ; Poonguzhali, Selvaraj ; Yim, Woo-Jong ; Indiragandhi, Pandiyan ; Kim, Kyoung-A ; Anandham, Rangasamy ; Yun, Jong-Chul ; Kim, Kye-Hoon ; Sa, Tongmin ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1622~1628
Bacteria from the Methylobacterium genus, called pink-pigmented facultative methylotrophic bacteria (PPFMs), are common inhabitants of plants, potentially dominating the phyllosphere population, and are also encountered in the rhizosphere, seeds, and other parts of plants, being versatile in nature. The consistent success of the Methylobacterium plant association relies on methylotrophy, the ability to utilize the one-carbon compound methanol emitted by plants. However, the efficiency of Methylobacterium in plant growth promotion could be better exploited and thus has attracted increasing interest in recent years. Accordingly, the present study investigated the inoculation effects of Methylobacterium sp. strains CBMB20 and CBMB 110 on seed imbibition to tomato and red pepper on the growth and accumulation of phytohormone levels under gnotobiotic conditions. Seeds treated with the Methylobacterium strains showed a significant increase in root length when compared with either the uninoculated control or Methylobacterium extorquens
knockout mutanttreated seeds. Extracts of the plant samples were used for indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), and dihydrozeatin riboside (DHZR) assays by immunoanalysis. The treatment with Methylobacterium sp. CBMB20 or CBMB 110 produced significant increases in the accumulation of IAA and the cytokinins t-ZR and DHZR in the red pepper extracts, whereas no IAA was detected in the tomato extracts, although the cytokinin concentrations were significantly increased. Therefore, this study proved that the versatility of Methylobacterium as a plant-growth promoting bacteria could be better exploited.
Differential Transformation of Ginsenosides from Panax ginseng by Lactic Acid Bacteria
Chi, Hyun ; Lee, Bo-Hyun ; You, Hyun-Ju ; Park, Myung-Soo ; Ji, Geun-Eog ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1629~1633
Ginsenosides have been regarded as the principal components responsible for the pharmacological and biological activities of ginseng. The transformation of ginsenosides with live lactic acid bacteria transformed ginsenosides Rb2 and Rc into Rd, but the reactions were slow. When the crude enzymes obtained from several lactic acid bacteria were used for transformation, those from Bifidobacterium sp. Int57 exhibited the most potent transforming activity of ginsenosides to compound K. In comparison, a relatively higher level of Rh2 was produced by the enzymes from Lactobacillus delbrueckii and Leuconostoc mesenteroides. These results suggest that it is feasible to develop a specific bioconversion process to obtain specific ginsenosides using the appropriate combination of ginsenoside substrates and specific microbial enzymes.
Inhibition of the Replication of Hepatitis C Virus Replicon with Nuclease-Resistant RNA Aptamers
Shin, Kyung-Sook ; Lim, Jong-Hoon ; Kim, Jung-Hye ; Myung, Hee-Joon ; Lee, Seong-Wook ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1634~1639
Hepatitis C virus (HCV)-encoded nonstructural protein 5B (NS5B) possesses RNA-dependent RNA polymerase activity, which is considered essential for viral proliferation. Thus, HCV NS5B is a good therapeutic target protein for the development of anti-HCV agents. In this study, we isolated two different kinds of nuclease-resistant RNA aptamers with 2'-fluoro pyrimidines against the HCV NS5B from a combinatorial RNA library with 40 nucleotide random sequences, using SELEX technology. The isolated RNA aptamers were observed to specifically and avidly bind the HCV NS5B with an apparent
of 5 nM and 18 nM, respectively, in contrast with the original RNA library that hardly bound the target protein. Moreover, these aptamers could partially inhibit RNA synthesis of the HCV subgenomic replicon when transfected into Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic agents of HCV infection but also as a powerful tool for the study of the HCV RNA-dependent RNA polymerase mechanism.
Characterization of the Microbial Diversity in a Korean Solar Saltern by 16S rRNA Gene Analysis
Park, Soo-Je ; Kang, Cheol-Hee ; Rhee, Sung-Keun ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1640~1645
We studied the diversity of the halophilic archaea and bacteria in crystallizer ponds of a Korean solar saltern by analyzing 16S rRNA gene libraries. Although diverse halophilic archaeal lineages were detected, the majority (56%) were affiliated with the uncultured and cultured Halorubrum group. Halophilic archaea that have been frequently observed in solar saltern environments previously, such as Halogeometricum, Halococcus, Haloarcula, and Haloferax, were not detected in our samples. The majority of clones (53%) belonged to the Cytophaga-Flavobacterium-Bacteroides and
groups, with 47% of the clones being affiliated with
. We also identified new
-related bacteria that have not been observed in hypersaline environments previously. Our data show that the diversity of the halophilic archaea and bacteria in our Korean saltern differs from that of solar salterns found in other geographic locations. We also showed by quantitative real-time PCR analysis that bacteria can form a significant component of the microbial community in solar salterns.
An In Vitro Assay to Screen for Translation Inhibitors
Song, Chin-Hee ; Paik, Hyoung-Rok ; Seong, Chi-Nam ; Choi, Sang-Ki ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1646~1649
Protein synthesis is the ultimate outcome of gene expression which, in turn, is regulated by several translation factors. We attempted to identify substances that can inhibit the translation process in vitro when the outcome protein is luciferase. To this end, we developed a sensitive cell-free protein synthesis assay using luciferase as the reporter. The synthesis of luciferase increased proportionately as mRNA was added to a
reaction medium in concentrations raging from 5 ng to 500 ng. The maximum amount of luciferase was synthesized when the media were incubated at
for 40 min. The concentration of each compound that inhibited luciferase production by 50% (
) was calculated. Hygromycin, puromycin, and cycloheximide yielded an
of 0.008, 0.8, and
, respectively. A filtrate of Streptomyces spp. isolates inhibited protein synthesis up to S-fold when added to the in vitro translation assay mixture.
Genome Snapshot of Paenibacillus polymyxa ATCC
Jeong, Hae-Young ; Kim, Ji-Hyun ; Park, Yon-Kyoung ; Kim, Seong-Bin ; Kim, Chang-Hoon ; Park, Seung-Hwan ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1650~1655
Bacteria belonging to the genus Paenibacillus are facultatively anaerobic endospore formers and are attracting growing ecological and agricultural interest, yet their genome information is very limited. The present study surveyed the genomic features of P. polymyxa ATCC
using pulse-field gel electrophoresis of restriction fragments and sample genome sequencing of 1,747 reads (approximately 17.5% coverage of the genome). Putative functions were assigned to more than 60% of the sequences. Functional classification of the sequences showed a similar pattern to that of B. subtilis. Sequence analysis suggests nitrogen fixation and antibiotic production by P. polymyxa ATCC
, which may explain its plant growth-promoting effects.
A Comparison of the Anti-inflammatory Activity of Surfactin A, B, C, and D from Bacillus subtilis
Kim, Sung-Dae ; Cho, Jae-Youl ; Park, Hwa-Jin ; Lim, Chang-Ryul ; Lim, Jong-Hwan ; Yun, Hyo-In ; Park, Seung-Chun ; Kim, Sang-Keun ; Rhee, Man-Hee ;
Journal of Microbiology and Biotechnology, volume 16, issue 10, 2006, Pages 1656~1659
Natural surfactins are a mixture of isoforms that differ slightly in their physiological properties. In previous research, we obtained surfactin A, B, C, and D from the Bacillus subtilis complex BC1212. We found that surfactin C inhibited nitric oxide (NO)-production and suppressed the expression of pro-inflammatory cytokine mRNA, which was stimulated by
of lipopolysaccharide (LPS) in murine RAW264.7 cells. In order to compare the anti-inflammatory effects of surf actin isoforms, we examined the inhibition of LPS-induced NO production and the pro-inflammatory cytokine expression level. Surfactin C inhibited the LPS-induced NO production in murine macrophage RAW264.7 cells the most. In addition, surf actin C was superior to other surfactin's subtypes regarding inhibiting the expression of inducible nitric oxide synthase (iNOS) and monocyte chemoattractant protein 1 (MCP-1). Finally, the anti-inflammatory activity of surf actin C is the most potent, compared with surfactin A, B, and D.