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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 24, Issue 5 - May 2014
Volume 24, Issue 4 - Apr 2014
Volume 24, Issue 3 - Mar 2014
Volume 24, Issue 2 - Feb 2014
Volume 24, Issue 1 - Jan 2014
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Advances in Rapid Detection Methods for Foodborne Pathogens
Zhao, Xihong ; Lin, Chii-Wann ; Wang, Jun ; Oh, Deog Hwan ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 297~312
DOI : 10.4014/jmb.1310.10013
Food safety is increasingly becoming an important public health issue, as foodborne diseases present a widespread and growing public health problem in both developed and developing countries. The rapid and precise monitoring and detection of foodborne pathogens are some of the most effective ways to control and prevent human foodborne infections. Traditional microbiological detection and identification methods for foodborne pathogens are well known to be time consuming and laborious as they are increasingly being perceived as insufficient to meet the demands of rapid food testing. Recently, various kinds of rapid detection, identification, and monitoring methods have been developed for foodborne pathogens, including nucleic-acid-based methods, immunological methods, and biosensor-based methods, etc. This article reviews the principles, characteristics, and applications of recent rapid detection methods for foodborne pathogens.
454 Pyrosequencing Analysis of Bacterial Diversity Revealed by a Comparative Study of Soils from Mining Subsidence and Reclamation Areas
Li, Yuanyuan ; Chen, Longqian ; Wen, Hongyu ; Zhou, Tianjian ; Zhang, Ting ; Gao, Xiali ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 313~323
DOI : 10.4014/jmb.1309.09001
Significant alteration in the microbial community can occur across reclamation areas suffering subsidence from mining. A reclamation site undergoing fertilization practices and an adjacent coal-excavated subsidence site (sites A and B, respectively) were examined to characterize the bacterial diversity using 454 high-throughput 16S rDNA sequencing. The dominant taxonomic groups in both the sites were Proteobacteria, Acidobacteria, Bacteroidetes, Betaproteobacteria, Actinobacteria, Gammaproteobacteria, Alphaproteobacteria, Deltaproteobacteria, Chloroflexi, and Firmicutes. However, the bacterial communities' abundance, diversity, and composition differed significantly between the sites. Site A presented higher bacterial diversity and more complex community structures than site B. The majority of sequences related to Proteobacteria, Gemmatimonadetes, Chloroflexi, Nitrospirae, Firmicutes, Betaproteobacteria, Deltaproteobacteria, and Anaerolineae were from site A; whereas those related to Actinobacteria, Planctomycetes, Bacteroidetes, Verrucomicrobia, Gammaproteobacteria, Nitriliruptoria, Alphaproteobacteria, and Phycisphaerae originated from site B. The distribution of some bacterial groups and subgroups in the two sites correlated with soil properties and vegetation due to reclamation practice. Site A exhibited enriched bacterial community, soil organic matter (SOM), and total nitrogen (TN), suggesting the presence of relatively diverse microorganisms. SOM and TN were important factors shaping the underlying microbial communities. Furthermore, the specific plant functional group (legumes) was also an important factor influencing soil microbial community composition. Thus, the effectiveness of 454 pyrosequencing in analyzing soil bacterial diversity was validated and an association between land ecological system restoration, mostly mediated by microbial communities, and an improvement in soil properties in coal-mining reclamation areas was suggested.
Diversity and Saline Resistance of Endophytic Fungi Associated with Pinus thunbergii in Coastal Shelterbelts of Korea
Min, Young Ju ; Park, Myung Soo ; Fong, Jonathan J. ; Quan, Ying ; Jung, Sungcheol ; Lim, Young Woon ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 324~333
DOI : 10.4014/jmb.1310.10041
The Black Pine, Pinus thunbergii, is widely distributed along the eastern coast of Korea and its importance as a shelterbelt was highlighted after tsunamis in Indonesia and Japan. The root endophytic diversity of P. thunbergii was investigated in three coastal regions; Goseong, Uljin, and Busan. Fungi were isolated from the root tips, and growth rates of pure cultures were measured and compared between PDA with and without 3% NaCl to determine their saline resistance. A total of 259 isolates were divided into 136 morphotypes, of which internal transcribed spacer region sequences identified 58 species. Representatives of each major fungi phylum were present: 44 Ascomycota, 8 Zygomycota, and 6 Basidiomycota. Eighteen species exhibited saline resistance, many of which were Penicillium and Trichoderma species. Shoreline habitats harbored higher saline-tolerant endophytic diversity compared with inland sites. This investigation indicates that endophytes of P. thunbergii living closer to the coast may have higher resistance to salinity and potentially have specific relationships with P. thunbergii.
Aspergillus cumulatus sp. nov., from Rice Straw and Air for Meju Fermentation
Kim, Dae-Ho ; Kim, Seon-Hwa ; Kwon, Soon-Wo ; Lee, Jong-Kyu ; Hong, Seung-Beom ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 334~336
DOI : 10.4014/jmb.1312.12006
A new species named Aspergillus cumulatus sp. nov. is described in Aspergillus section Aspergillus (Eurotium state). The type strain (KACC
) of this species was isolated from rice straw used in meju fermentations in Korea, and other strains were isolated from the air in a meju fermentation room. The species is characterized by growth at a wide range of water activities and the formation of aerial hyphae on malt extract 60% sucrose agar (ME60S) that resemble a cumulus cloud. Furthermore, A. cumulatus produces yellow ascomata containing small lenticular ascospores (5.1-5.7
) with a wide furrow, low equatorial crests, and tuberculate convex surface. The species is phylogenetically distinct from the other reported Aspergillus section Aspergillus species based on multilocus sequence typing using rDNA-ITS,
-tubulin, calmodulin, and RNA polymerase II genes.
A New Signal Sequence for Recombinant Protein Secretion in Pichia pastoris
Govindappa, Nagaraj ; Hanumanthappa, Manjunatha ; Venkatarangaiah, Krishna ; Periyasamy, Sankar ; Sreenivas, Suma ; Soni, Rajeev ; Sastry, Kedarnath ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 337~345
DOI : 10.4014/jmb.1308.08085
Pichia pastoris is one of the most widely used expression systems for the secretory expression of recombinant proteins. The secretory expression in P. pastoris usually makes use of the prepro
sequence from Saccharomyces cerevisiae, which has a dibasic amino acid cleavage site at the end of the signal sequence. This is efficiently processed by Kex2 protease, resulting in the secretion of high levels of proteins to the medium. However, the proteins that are having the internal accessible dibasic amino acids such as KR and RR in the coding region cannot be expressed using this signal sequence, as the protein will be fragmented. We have identified a new signal sequence of 18 amino acids from a P. pastoris protein that can secrete proteins to the medium efficiently. The PMT1-gene-inactivated P. pastoris strain secretes a ~30 kDa protein into the extracellular medium. We have identified this protein by determining its N-terminal amino acid sequence. The protein secreted has four DDDK concatameric internal repeats. This protein was not secreted in the wild-type P. pastoris under normal culture conditions. We show that the 18-amino-acid signal peptide at the N-terminal of this protein is useful for secretion of heterologous proteins in Pichia.
Glioblastoma-Specific Anticancer Activity of Pheophorbide a from the Edible Red Seaweed Grateloupia elliptica
Cho, MyoungLae ; Park, Gab-Man ; Kim, Su-Nam ; Amna, Touseef ; Lee, Seokjoon ; Shin, Woon-Seob ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 346~353
DOI : 10.4014/jmb.1308.08090
The chlorophyll-related compound pheophorbide a (Pa) was successively purified from an edible red seaweed, Grateloupia elliptica, using silica, octadecyl silica column chromatography and reversed phase-high-performance liquid chromatography, as well as the cell cycle inhibitory and apoptotic effects of Pa being investigated in U87MG glioblastoma cells. The Pa exhibited strong anticancer effects in the absence of direct photo-irradiation against various cancer cell lines, including U87MG, SK-OV-3, and HeLa cells. Among the cancer cells, the strongest anticancer activity of Pa exhibited on U87MG cells with
values of 2.8
. In addition, Pa specifically had cytostatic activity on glioblastoma cells rather than human umbilical vein endothelial cells. Analysis of the cell cycle distribution showed that Pa induced G0/G1 arrest of U87 MG cells. In addition, arrested cells induced late apoptosis and DNA degradation under dark condition. These results suggest that Pa isolated from G. elliptica is a potential glioblastoma-specific anticancer agent without side effects on normal cells.
Lycorine: A Potential Broad-Spectrum Agent Against Crop Pathogenic Fungi
Shen, Jin-Wen ; Ruan, Yuan ; Ren, Wei ; Ma, Bing-Ji ; Wang, Xiao-Long ; Zheng, Chun-Feng ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 354~358
DOI : 10.4014/jmb.1310.10063
A screening test showed that lycorine exhibited significant antifungal activity against 24 pathogenic crop fungi at concentrations of 500
, respectively. Fusarium graminearum was selected for antifungal mechanism studies by observing its mycelial morphology and investigating the variations in its conductivity. In addition, the substance absorption and metabolism of F. graminearum were explored. The mechanism was revealed as being one by which lycorine destroyed the cellular membrane and further influenced substance absorption and cell metabolism.
Enzymatic Production of 15-Hydroxyeicosatetraenoic Acid from Arachidonic Acid by Using Soybean Lipoxygenase
Kim, Baek-Joong ; Shin, Kyung-Chul ; Oh, Deok-Kun ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 359~362
DOI : 10.4014/jmb.1310.10054
15-Hydroxyeicosatetraenoic acid (HETE), as a mammalian biologically active metabolite, has anticarcinogenic effect. The conditions of producing 15-HETE from arachidonic acid by using soybean lipoxygenase were optimal at pH 8.5 and
with 9 g/l arachidonic acid, 54.4 U/ml soybean lipoxygenase, and 4% methanol. Under these optimized conditions, the enzyme produced 9.5 g/l 15-HETE after 25 min, with a molar conversion yield of 99% and a productivity of
. To the best of our knowledge, this is the first biotechnological production of 15-HETE.
Use of In Vivo-Induced Antigen Technology to Identify In Vivo-Expressed Genes of Campylobacter jejuni During Human Infection
Hu, Yuanqing ; Huang, Jinlin ; Li, Qiuchun ; Shang, Yuwei ; Ren, Fangzhe ; Jiao, Yang ; Liu, Zhicheng ; Pan, Zhiming ; Jiao, Xin-An ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 363~370
DOI : 10.4014/jmb.1311.11019
Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.
Protective Effects of Probiotic Lactobacillus rhamnosus IMC501 in Mice Treated with PhIP
Dominici, Luca ; Villarini, Milena ; Trotta, Francesca ; Federici, Ermanno ; Cenci, Giovanni ; Moretti, Massimo ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 371~378
DOI : 10.4014/jmb.1309.09072
The aim of the present study was to investigate the antigenotoxic properties of the probiotic Lactobacillus rhamnosus IMC501; DNA damage was induced by one representative food mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Mice were treated orally with suspensions of lactobacilli for 10 days before administration of food mutagen. During the treatment, the abundance of lactobacilli in feces, as assessed by qPCR analysis, increased, whereas
-glucuronidase and N-acetyl-
-glucosaminidase activities decreased. The extent of DNA damage was measured in colon and liver cells by comet assay. In colonocytes, diet supplementation with IMC501 resulted in a significant inhibition of DNA damage induced by PhIP. The results obtained in this in vitro study suggest that Lactobacillus rhamnosus IMC501 used as a dietary supplement can provide a useful integration of antimutagen food components of the normal diet, which are generally lower than the protective level.
Comparison of the Organophosphorus Hydrolase Surface Display Using InaVN and Lpp-OmpA Systems in Escherichia coli
Karami, Ali ; Latifi, Ali Mohamad ; Khodi, Samaneh ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 379~385
DOI : 10.4014/jmb.1309.09066
The purpose of this study was to compare the ability of an engineered Escherichia coli to degrade chlorpyrifos (Cp) using an organophosphorus hydrolase enzyme, encoded in both Flavobacterium sp. ATCC 27551 or Pseudomonas diminuta, by employing the Lpp-OmpA chimera and the N-terminal domain of the ice nucleation protein as anchoring motifs. Tracing of the expression location of the recombinant protein using SDS-PAGE showed the presentation of OPH by both anchors on the outer membrane. This is the first report on the presentation of OPH on the cell surface by Lpp-OmpA under the control of the T7 promoter. The results showed cell growth in the presence of Cp as the sole source of energy, without growth inhibition, and with higher whole-cell activity for both cells harboring plasmids pENVO and pELMO, at approximately 10,342.85 and 10,857.14 U/mg, respectively. Noticeably, the protein displayed by pELMO was lower than the protein displayed by pENVO. It can be concluded that Lpp-OmpA can display less protein, but more functional OPH protein. These results highlight the high potential, of both engineered bacteria, for use in the bioremediation of pesticide-contaminated sources in the environment.
Characterization of Veterinary Hospital-Associated Isolates of Enterococcus Species in Korea
Chung, Yeon Soo ; Kwon, Ka Hee ; Shin, Sook ; Kim, Jae Hong ; Park, Yong Ho ; Yoon, Jang Won ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 386~393
DOI : 10.4014/jmb.1310.10088
Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n=465) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.
Effect of Lipopolysaccharide (LPS) on Mouse Model of Steroid-Induced Avascular Necrosis in the Femoral Head (ANFH)
Ryoo, Soyoon ; Lee, Sukha ; Jo, Seunghyun ; Lee, Siyoung ; Kwak, Areum ; Kim, Eunsom ; Lee, Jongho ; Hong, Jaewoo ; Jhun, Hyunjhung ; Lee, Youngmin ; Sobti, Anshul Shyam ; Kim, Soohyun ; Oh, Kwang-Jun ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 394~400
DOI : 10.4014/jmb.1311.11057
Avascular necrosis of the femoral head (ANFH) is commonly observed in patients treated with excessive glucocorticoid (GC). Single administration of lipopolysaccharide (LPS) has shown to induce immune stimulatory factors. However, the effect of repeated administration of LPS on GC-induced ANFH has not been studied. Thus, the purpose of this study was (i) to examine the cytokine profile induced by repeated LPS administrations and (ii) to test the effect of repeated LPS treatments on GC-induced ANFH. A mouse necrosis model of ANFH was designed by chronic GC administration with co-treatment of LPS. Mice body weights in the LPS/prednisolone (PDN) co-treated group were lower than that of the untreated control group, but spleen weights were greater than the control group. The levels of IL-6,
, and IL-33 in the liver and spleen of the LPS/PDN group were lower than the untreated control group, whereas
level in the femoral head of the LPS/PDN group increased. Collectively, the effect of repeated LPS on the pathogenesis of GC-induced ANFH was associated with the
level in the femoral head, but the pathogenesis did not correspond to cytokine levels in immune tissues.
Interaction Between the Quorum Sensing and Stringent Response Regulation Systems in the Enterohemorrhagic Escherichia coli O157:H7 EDL933 Strain
Oh, Kyung-Hwan ; Cho, Seung-Hak ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 401~407
DOI : 10.4014/jmb.1310.10091
Quorum sensing and the stringent response are well-known regulation systems for the expression of virulence genes in enterohemorrhagic Escherichia coli (EHEC). However, how these two systems interact is not well known. E. coli strains with mutations in two regulation systems,
(ECM201), and the
complement strain to ECM201 (ECM202) were created from EHEC O157:H7 EDL933 to investigate how the regulatory systems interact. The phenotypic changes of the mutant strains were characterized and compared with the wild type. The mutant strains exhibited no obvious growth defects, although acid resistance and cellular cytotoxicity were decreased significantly in all the mutant strains. Phenotypic characterization revealed that mutations in the stringent response system (ECM201 and ECM202) influenced the metabolic (defective utilization of arabinose and L-sorbose) and enzymatic activities (decreased trypsin activity, and increased
-glucosidase activity). In contrast, the quorum sensing system mutant (ECM101) did not display these phenotypes. The motility of the quorum sensing system mutant (ECM101) was unchanged, but mutation in the stringent response system influenced the motility. Our results suggest that quorum sensing interacts with the stringent response regulation system.
Construction of a Large Synthetic Human Fab Antibody Library on Yeast Cell Surface by Optimized Yeast Mating
Baek, Du-San ; Kim, Yong-Sung ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 408~420
DOI : 10.4014/jmb.1401.01002
Yeast surface-displayed antibody libraries provide an efficient and quantitative screening resource for given antigens, but suffer from typically modest library sizes owing to low yeast transformation efficiency. Yeast mating is an attractive method for overcoming the limit of yeast transformation to construct a large, combinatorial antibody library, but the optimal conditions have not been reported. Here, we report a large synthetic human Fab (antigen binding fragment) yeast surface-displayed library generated by stepwise optimization of yeast mating conditions. We first constructed HC (heavy chain) and LC (light chain) libraries, where all of the six CDRs (complementarity-determining regions) of the variable domains were diversified mimicking the human germline antibody repertoires by degenerate codons, onto single frameworks of VH3-23 and
-16 germline sequences, in two haploid cells of opposite mating types. Yeast mating conditions were optimized in the order of cell density, media pH, and cell growth phase, yielding a mating efficiency of ~58% between the two haploid cells carrying HC and LC libraries. We constructed two combinatorial Fab libraries with CDR-H3 of 9 or 11 residues in length with colony diversities of more than
by one round of yeast mating between the two haploid HC and LC libraries, with modest diversity sizes of
. The synthetic human Fab yeast-displayed libraries exhibited relative amino acid compositions in each position of the six CDRs that were very similar to those of the designed repertoires, suggesting that they are a promising source for human Fab antibody screening.
Characterization of CTX-M-Type Extended-Spectrum Beta-Lactamase-Producing Diarrheagenic Escherichia coli Isolates in the Republic of Korea During 2008-2011
Kim, Jin Seok ; Kim, Junyoung ; Kim, Soo-Jin ; Jeon, Se-Eun ; Oh, Kyung Hwan ; Cho, Seung-Hak ; Kang, Yeon-Ho ; Han, Soon Young ; Chung, Gyung Tae ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 421~426
DOI : 10.4014/jmb.1401.01023
To characterize the extended-spectrum beta-lactamases (ESBLs) in diarrheagenic Escherichia coli from Korea in 2008-2011, we screened seven enterotoxigenic E. coli (ETEC) and one enteroaggregative E. coli (EAEC) that produce ESBLs from a nationwide survey. All eight isolates produced CTX-M-type ESBLs, including CTX-M-12 (n = 4), CTX-M-14 (n = 2), and CTX-M-15 (n = 2). PCR-based replicon typing indicated that the
genes of four ETEC isolates were carried on a conjugative IncF plasmid, whereas the
of one EAEC was located on an IncK plasmid. This is the first report of the occurrence of
genes in clinical isolates of EAEC in Korea. The ESBL-producing isolates were shown to be different based on pulsed-field gel electrophoresis and multilocus sequence typing, whereas the four isolates with CTX-M-12 were clonally related. These observations raise an alarm for the spread of plasmid-mediated resistance to ESBL among diarrheagenic E. coli.
Development of a Novel Immunochromatographic Assay for Rapid Detection of VanA Ligase-Producing Vancomycin-Resistant Enterococci
Ji, Gil Yong ; Song, Hyung Geun ; Son, Bo Ra ; Hong, Seung Bok ; Kim, Jong Wan ; Shin, Kyeong Seob ;
Journal of Microbiology and Biotechnology, volume 24, issue 3, 2014, Pages 427~430
DOI : 10.4014/jmb.1307.07035
We developed a novel immunochromatographic assay (ICA) (EZ-Step VanA rapid kit; Dinona, Korea) for the detection of VanA ligase from vancomycin-resistant enterococci (VRE). Of eight monoclonal antibodies screened by ELISAs, the VanA ligase ICA constructed with 1H9 plus 3G11 showed the greatest reactivity. The detection limit of the kit was
CFU per test. Of 127 vancomycin-resistant microorganisms, 100 vanA VRE were positive in the VanA ligase ICA, and 27 non-vanA vancomycin-resistant isolates were negative. These results were consistent with those of the PCR analyses. Thus, our ICA is a reliable and easy-to-use immunological assay for detecting VanA-producing VRE in clinical laboratories.