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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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The Microbiological Society of Korea
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Volume & Issues
Volume 52, Issue 2 - Jun 2016
Volume 52, Issue 1 - Mar 2016
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A unique thioredoxin reductase plays defensive roles against oxidative, nitrosative and nutritional stresses in Schizosaccharomyces pombe
Ji, Dam-Jung ; Lim, Chang-Jin ; Kim, Kyunghoon ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 1~9
DOI : 10.7845/kjm.2016.6004
A unique Schizosaccharomyces pombe
gene encoding thioredoxin reductase (TrxR) was found to be positively regulated by stress-inducing agents through the stress-responsive transcription factor Pap1. In the present study, the protective roles of S. pombe TrxR were evaluated using the TrxR-overexpressing recombinant plasmid pHSM10. In the presence of hydrogen peroxide (
) and superoxide anion-generating menadione (MD), S. pombe TrxR increased cellular growth and the total glutathione (GSH) level, while it reduced levels of intracellular reactive oxygen species (ROS). The nitric oxide (NO) levels of the TrxR-overexpressing cells, in the presence of
and MD, were maintained to be similar to those of the corresponding non-treated cells. Although S. pombe TrxR was able to scavenge NO generated by sodium nitroprusside (SNP), it had no significant modulating effects on cellular growth, ROS levels, or the total GSH level of SNP-exposed yeast cells, compared with the differences in those of the two non-treated cell cultures. TrxR increased the cellular growth and total GSH level, which were diminished by nitrogen starvation. It also scavenged ROS and NO produced during nitrogen starvation. Taken together, the S. pombe TrxR protects against oxidative, nitrosative, and nutritional stresses.
The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Escherichia coli alkaline phosphatase
Han, Seung Hee ; Kim, Jin-Kyoo ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 10~17
DOI : 10.7845/kjm.2016.6008
Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor
related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.
Anti-allergic effect of Lactobacillus rhamnosus IDCC 3201 isolated from breast milk-fed Korean infant
Lee, Seung-Hun ; Kang, Jae-Hoon ; Kang, Dae-Jung ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 18~24
DOI : 10.7845/kjm.2016.6013
We investigated 23 lactic acid bacteria isolated from Korean breast milk-fed infant in order to select strains which show superior anti-allergic effect. The candidates were cultivated and then we obtained dried powders of tyndallized cells and supernatant concentrate separately. Screening was carried out with down-regulation of interleukin (IL) 4 and up-regulation of IFN-
in mouse splenocytes. As a result of the screening, we selected Lactobacillus rhamnosus IDCC 3201 (RH3201) for oral feeding to ovalbumin-sensitized BALB/c mice. Oral administration of RH3201 as dead cell bodies and supernatant concentrate suppressed hyper-production of serum immunoglobulin (Ig) E levels compared to vehicle group. Such anti-allergic effects were achieved by improvement of the balance between cytokines produced from type-1 helper T (Th1) and type-2 helper T (Th2) lymphocytes. Therefore, RH3201 has potential to improve atopic symptoms by immunomodulatory effect.
Development of selective media for Enterococci
Chang, Dong-Ho ; Yoon, Jun-Beom ; Lee, Keun Heon ; Park, Kyeong Ryang ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 25~31
DOI : 10.7845/kjm.2016.5068
An enterococci selective medium was developed to detect the presence of enterococci for use as a fecal contamination indicator. Among several media which have been known to detect enterococci, the following 9 different kinds of media were selected: Enterococci Confirmatory agar, Azide dextrose agar, Bromocresol-purple azide agar, Esculin bile agar, Citrate azide tween carbonate agar, KF Streptococcus agar, BROLACIN agar, Kanamycin esculin azide agar, and Membrane filter Enterococcus selective agar. Various components from the nine media were mixed to develop a more effective enterococcus selective medium. The newly developed medium named as `Enterococcus Mixed medium` was more effective than the previous 9 media. Enterococci strains (Enterococcus avium KACC 10788, Enterococcus faecium KACC 11954, Enterococcus saccharolyticus KACC 10783, Enterococcus durans KACC 10787, Enterococcus faecalis KACC 11304, and Enterococcus hirae KACC 10779) and non-enterococci strains (Escherichia coli KACC 10005, Staphylococcus aureus subsp. aureus KACC 10768, and Bacillus subtilis KACC 10111) were used to test the new medium. As a result, the enterococci strains grew well on the Enterococcus Mixed medium whereas the non-enterococci strains did not grow well on it. Additionally, growth of enterococci with freshwater and seawater samples was observed to be good on the Enterococcus Mixed medium. The result of this study confirmed that the Enterococcus Mixed medium was effective in detecting the target enterococci.
Gibberellin A7 production by Aspergillus tubingensis YH103 and cultural characteristics of endophytic fungi isolated from Tetragonia tetragonoides in Dokdo islands
You, Young-Hyun ; Park, Jong Myong ; Lim, Sung Hwan ; Kang, Sang-Mo ; Park, Jong-Han ; Lee, In-Jung ; Kim, Jong-Guk ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 32~39
DOI : 10.7845/kjm.2016.5071
Coastal plant species Tetragonia tetragonoides (Pall.) Kuntze native to the Dokdo islands was sampled and then 17 endophytic fungi were purely isolated based on morphological differences. The fungal isolates were characterized by their growth properties under NaCl concentration or pH gradient. Culture filtrates of the 17 fungal isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity. As the results, YH103 strain showed the highest plant growth-promoting activity among them. Phylogenetic analysis of the isolates was done by the maximum likelihood method based on partial internal transcribed spacer region (ITS region: contaning ITS1, 5.8S, and ITS2), beta-tubulin (BenA), and calmodulin (CaM) gene sequences. Chromatographic analysis of the strain YH103 culture filtrate showed the existence of gibberellins (
). Finally, the strain YH103 was identified as Aspergillus tubingensis by microscopic observation and molecular analysis and, to our knowledge, this is the first report of GAs producing A. tubingensis.
Isolation of marine algicidal bacteria from surface seawater and sediment samples associated with harmful algal blooms in Korea
Kristyanto, Sylvia ; Kim, Jaisoo ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 40~48
DOI : 10.7845/kjm.2016.5048
This study mainly focused on isolation of marine algicidal bacteria associated with phytoplankton blooms and characterization of algicidal activity against harmful algae. Harmful algal blooms (HABs) found naturally in surface waters have caused many environmental problems worldwide. In this study, forty bacterial strains that have capability of inhibiting harmful algal growth were isolated from Masan Bay, Jinhae Bay, Dol Island, Jangmok Bay, and the Tongyeong Sea, Republic of Korea. The bacteria were screened furthermore for the characteristics on algicidal activities against Cochlodinium polykrikoides, Chattonella marina, Skeletonema costatum, Heterosigma akashiwo, Heterocapsa triquetra, Prorocentrum minimum, and Scrippsiella trochoidea. As a result, the algicidal bacteria that were screened from double over layer agar and microscopic counts tests belonged to genera Pseudomonas, Vibrio, Bacillus, Pseudoalteromonas, Ruegeria, Joostella, Marinomonas, Stakelama, Porphyrobacter, and Albirhodobacter. One of the most important HAB species is Co. polykrikoides and the strongest algicidal activity against the dinoflagellate was 94.00% after 6 h treatment with 10% bacterial culture filtrate. In this study, Marinomonas sp. M Jin 1-8, Stakelama sp. ZB Yeonmyeong 1-11 & 1-13, Porphyrobacter sp. M Yeonmyeong 2-22, and Albirhodobacter sp. 6-R Jin 6-1 were found to be as new genera of bacteria having anti-algal activity. These results suggest that these bacteria might play an important role in controlling phytoplankton blooms.
Succession of bacterial community structure during the early stage of biofilm development in the Antarctic marine environment
Lee, Yung Mi ; Cho, Kyung Hee ; Hwang, Kyuin ; Kim, Eun Hye ; Kim, Mincheol ; Hong, Soon Gyu ; Lee, Hong Kum ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 49~58
DOI : 10.7845/kjm.2016.6005
Compared to planktonic bacterial populations, biofilms have distinct bacterial community structures and play important ecological roles in various aquatic environments. Despite their ecological importance in nature, bacterial community structure and its succession during biofilm development in the Antarctic marine environment have not been elucidated. In this study, the succession of bacterial community, particularly during the early stage of biofilm development, in the Antarctic marine environment was investigated by pyrosequencing of the 16S rRNA gene. Overall bacterial distribution in biofilms differed considerably from surrounding seawater. Relative abundance of Gammaproteobacteria and Bacteroidetes which accounted for 78.9-88.3% of bacterial community changed drastically during biofilm succession. Gammaproteobacteria became more abundant with proceeding succession (75.7% on day 4) and decreased to 46.1% on day 7. The relative abundance of Bacteroidetes showed opposite trend to Gammaproteobacteria, decreasing from the early days to the intermediate days and becoming more abundant in the later days. There were striking differences in the composition of major OTUs (
) among samples during the early stages of biofilm formation. Gammaproteobacterial species increased until day 4, while members of Bacteroidetes, the most dominant group on day 1, decreased until day 4 and then increased again. Interestingly, Pseudoalteromonas prydzensis was predominant, accounting for up to 67.4% of the biofilm bacterial community and indicating its important roles in the biofilm development.
Phylogenetic characteristics of actinobacterial population in bamboo (Sasa borealis) soil
Lee, Hyo-Jin ; Han, Song-Ih ; Whang, Kyung-Sook ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 59~64
DOI : 10.7845/kjm.2016.6006
In this study, a pyrosequencing was performed and analyzed to verify the phylogenetic diversity of actinomycetes in the bamboo (Sasa borealis) soil as a base study to obtain the genetic resources of actinomycetes. It was found that the rhizosphere soil had much various distribution in bacterial communities showing a diversity of 8.15 with 2,868 OTUs, while the litter layer showed a diversity of 7.55 with 2,588 OTUs. The bacterial community in the bamboo soil was composed of 35 phyla and the predominant phyla were Proteobacteria (51-60%), Bacteroidetes (16-20%), Acidobacteria (4-16%) and Actinobacteria (4-14%). In particular, Actinobacteria including Micromonosporaceae and Streptomycetaceae had a diverse distribution of actinomycetes within the six orders, 35 families and 121 genera, and it was characterized that about 83% of actinomycetes within Actinomycetales belonged to the 28 families. Among the dominant actinobacterial populations, Micromonosporaceae, Pseudonocardiaceae and Streptomycetaceae were representative family groups in the bamboo soils.
Phylogenetic diversity and UV resistance analysis of radiation-resistant bacteria isolated from the water in Han River
Lee, Jae-Jin ; Joo, Eun Sun ; Lee, Do Hee ; Jung, Hee-Young ; Kim, Myung Kyum ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 65~73
DOI : 10.7845/kjm.2016.6015
The aim of this study was to investigate the UV-resistance of radiation-resistant bacteria isolated from the water of Han River, South Korea. The water sample was irradiated with 3 kGy gamma radiation prior to isolation. Radiation-resistant bacterial strains were isolated by standard serial dilution method on R2A and 1/10 diluted R2A agar. The resulting purely isolated 60 cultures of bacteria were analysed for UV resistance and used in further studies. Based on the comparative analyses of 16S rRNA gene sequences, the bacterial isolates were divided into 3 phyla (4 genera): the phylum Deinococcus-Thermus (the genus Deinococcus) was 61.7%, Bacteroidetes (Hymenobacter and Spirosoma) was 23.4%, and Firmicutes (Exiguobacterium) was 15%. The results suggested that twenty-nine isolates are candidates new species belonging to Deinococcus, Hymenobacter, and Spirosoma, or other new genera. Nine bacterial strains were selected among the novel candidates and the UV-resistance analysis was conducted. All the candidate bacterial strains showed high UV resistance, similar to that of D. radiodurans R1.
Characteristic study and isolation of Bacillus subtilis SRCM 101269 for application of cow manure
Jeon, SaeBom ; Oh, HyeonHwa ; Uhm, Tai-Boong ; Cho, Jae-Young ; Yang, Hee-Jong ; Jeong, Do-Youn ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 74~83
DOI : 10.7845/kjm.2016.6001
Bacillus subtilis SRCM 101269 having safety and amo gene isolated from Korean traditional fermented food and their investigated characterization to apply the cow manure such as cellulase and xylanase activities, 16S rRNA sequencing, and ability of removal of livestock manure odor. Cow manure application results for the removal of livestock manure odor, the ammonia gas was reduced more than two-folder compared to the control group after 6 days, and reduced to less than 10 ppm after 9 days. In the case of cow manure added fowl droppings and other wood-based mixture components, ammonia gas maintained constant after 3 days of fermentation. However, in the case of sample inoculated B. subtilis SRCM 101269, ammonia gas reduced in course of fermentation time, and concentration of hydrogen sulfide also reduced for 65 ppm. Changes of nitrite concentration according to fermentation time no showed different for cow manure, however nitrite concentration in mixed livestock manure increased when compared to control. And then sulfate concentration in cow manure decreased, and no showed different when compared to the initial fermentation. No apparent change of sulfate concentration in mixed livestock manure detected. Through the previously studies, B. subtilis SRCM 101269 has high potential in industrial application manufacturing the cow manure as removal of livestock manure odor.
Microbiological and chemical properties of sourdough fermented with probiotic lactic acid bacteria
Lim, Eun-Seo ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 84~97
DOI : 10.7845/kjm.2016.6012
Isolates from Korean fermented soybean paste were identified as Enterococcus faecium SBP12, Pediococcus halophilus SBP20, Lactobacillus fermentum SBP33, Leuconostoc mesenteroides SBP37, Pediococcus pentosaceus SBP41, Lactobacillus brevis SBP49, Lactobacillus acidophilus SBP55, and Enterococcus faecalis SBP58 according to conventional morphological and biochemical characteristics, carbohydrate fermentation profiling, and 16S rRNA sequence comparison. Strain SBP20, SBP33, SBP49, and SBP55 showed very resistance to simulated gastric and intestinal juices with final populations exceeding 6 log CFU/ml, whereas cells of SBP12 and SBP58 after exposure to low pH were dramatically decreased within 2 h. Among 4 strains having good tolerance to gastrointestinal conditions, the high adhesive ability to HT-29 cells, antibiotic resistance, and antimicrobial activity against food-borne pathogens Bacillus cereus ATCC 11778 and Staphylococcus aureus ATCC 6538 were observed with SBP49 and SBP55, therefore, these two strains were confirmed as putative probiotic candidates. There was no significant difference between the sourdoughs fermented with SBP49 and SBP55 with respect to the values of pH, total titratable acidity, and viable cell count. During sourdough fermentation, SBP49 strain produced significantly greater amounts of lactic acid than SBP55 strain, which secreted large quantities of hydrogen peroxide. SBP49 and SBP55 strains producing the antimicrobial substances such as lactic acid, hydrogen peroxide, and bacteriocin effectively inhibited B. cereus and S. aureus inoculated in the sourdough.
Physicochemical characteristics and volatile flavor compounds of produced mixture wine with kiwi and permission fruits using wild yeast, Saccharomyces cerevisiae Y28
Lee, Hee Yul ; Seo, Weon Taek ; Jeong, Seong Hoon ; Hwang, Chung Eun ; Ahn, Min Ju ; Lee, Ae Ryeon ; Shin, Ji Hyun ; Lee, Joo Young ; Jo, Hyeon Kook ; Cho, Kye Man ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 98~109
DOI : 10.7845/kjm.2016.5072
The study was aimed to investigate the mixing ratio of kiwi and persimmon juices for the production of good quality wine by Saccharomyces cerevisiae Y28. Firstly, the optimum condition of rapidase treatment for the kiwi and persimmon juices was established, thereafter various mixing ratio (10:0, 9:1, 8:2, 7:3, 6:4, 5:5) of kiwi and persimmon was investigated regarding physiochemical properties and flavor compounds of wine. As the result, the optimum conditions were obtained as 0.3% rapidase for 1 h in kiwi and 0.3% rapidase for 3 h in persimmon. According to higher ration of persimmon, the pH of wines increased from 3.69 to 3.77, while the acidity of wines decreased from 2.07% to 1.51% at 14 days fermentation. The ranges of brix and reducing sugar in wines were decreased which ranges around 9.6 to 8.8 and 6.07 to 6.90 g/L, respectively, after fermentation. Major organic acid in wines were identified as tartaric acid, malic acid, and citric acid. A small amount of free sugar such as sucrose and glucose were detected in wines, but fructose was completely absent. The soluble phenolic contents were decreased that ranges around 1.00 to 1.25 g/L, in contrast, browning degree were increased ranges around 0.212 to 0.412 after fermentation. The major flavor components were identified as ethyl acetate and hydrazine, and 1,1-dimethyl. Importantly, phenylethyl alcohol was detected from the all wines that have a typical rose like flavor. But sensory test results and preference of kiwi-persimmon (7:3) mixing wine was better than the other wines.
Construction of Pseudoalteromonas - Escherichia coli shuttle vector based on a small plasmid from the marine organism Pseudoalteromonas
Kim, Dockyu ; Park, Ha Ju ; Park, Hyun ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 110~115
DOI : 10.7845/kjm.2016.5054
A small plasmid (pDK4) from the Antarctic marine organism Pseudoalteromonas sp. PAMC 21150, was purified, sequenced and analyzed. pDK4 was determined to be 3,480 bp in length with a G+C content of 41.64% and contains three open reading frames encoding a replication initiation protein (RepA), a conjugative mobilization protein (Mob) and a hypothetical protein. PCR-amplified pDK4 was cloned in high-copy pUC19 to yield the fusion vector pDOC153. The chloramphenicol resistance gene was inserted into pDOC153 to give an ampicillin and chloramphenicol-resistant, Pseudoalteromonas - Escherichia coli shuttle vector (7,216 bp; pDOC155). The TonB-dependent receptor (chi22718_IV ) and exochitinase (chi22718_III ) genes from Arctic marine P. issachenkonii PAMC 22718 were cloned into pDOC155 to produce pDOC158 and pDOC165, respectively. Both vector derivatives were transferred into plasmid-free Pseudoalteromonas sp. PAMC 22137 by the triparental mating method. PCR experiments showed that the genes were stably maintained both in Pseudoalteromonas sp. PAMC 22137 and E. coli
cells, indicating the potential use of pDOC155 as a new gene transfer system into marine Pseudoalteromonas spp.
Development of a novel genetic assay for telomere recombination in Saccharomyces cerevisiae
Kim, Min-Kyu ; Bae, Sung-Ho ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 116~119
DOI : 10.7845/kjm.2016.5070
Stable maintenance of telomere is required for cell proliferation and survival. Although telomerase is the primary means for telomere maintenance, recombination is another important pathway to maintain telomeres. In this study, we developed a genetic assay for telomere recombination using the internal
repeats present in subtelomeric regions of yeast. The recombination frequencies were dependent on the presence of the internal
repeats. PCR amplification of the regions near URA3 and CAN1 markers using genomic DNA isolated from
colonies indicated that each isolate had lost the chromosome end including the markers. In addition, the recombination frequencies increased with longer internal
repeats. Our results suggest that the
colony formation is the consequence of recombination between the internal and terminal
Characterization of the bacteriophage P4 sid
derivative overcoming P2sir-associated helper inefficiency through DNA conformational adaptation
Kim, Kyoung-Jin ;
The Korean Journal of Microbiology, volume 52, issue 1, 2016, Pages 120~124
DOI : 10.7845/kjm.2016.6009
A certain size of DNA (28-29 kb long) to be packaged into P2-size head and the mutation in sid gene of bacteriophage P4 are the major factors to overcome "P2 sir-associated helper inefficiency". To clarify whether the presence of sid mutation is essential to overcome "P2 sir-associated helper inefficiency" or not, we tested the P4 derivative, P4 delRI::kmr, which is
and whose genome size supposed to be 28.5 kb long in the case of being packaged into
-sized large head. As P4 delRI::kmr showed the low EOP with P2 sir3 lysogen, P4 delRI::kmr phage stock was prepared in P2 sir3 lysogen host to increase the EOP with P2 sir3 lysogen. Through this process, P4 delRI::kmr had been adapted for P2 sir3 lysogen. With a CsCl buoyant equilibrium density gradient experiment and gel electrophoresis of the isolated DNA, it was evident that the adaptation of P4 delRI::kmr for P2 sir3 lysogen was caused by the conformational change of DNA to be packaged into large head. The burst size determination experiments with P4 delRI::kmr phage stock adapted for P2 sir3 lysogen and normal P4 delRI::kmr phage stock showed that not the sid mutation but the size of DNA to be packaged (28-29 kb long) was essential to overcome "P2 sir-associated helper inefficiency".