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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
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Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 46, Issue 4 - Dec 2010
Volume 46, Issue 3 - Sep 2010
Volume 46, Issue 2 - Jun 2010
Volume 46, Issue 1 - Mar 2010
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Metaproteomics in Microbial Ecology
Kim, Jong-Shik ; Woo, Jung-Hee ; Kim, Jun-Tae ; Park, Nyun-Ho ; Kim, Choong-Gon ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 1~8
New technologies are providing unprecedented knowledge into microbial community structure and functions. Even though nucleic acid based approaches provide a lot of information, metaproteomics could provide a high-resolution representation of genotypic and phenotypic traits of distinct microbial communities. Analyzing the metagenome from different microbial ecosystems, metaproteomics has been applied to seawater, human guts, activated sludge, acid mine drainage biofilm, and soil. Although these studies employed different approaches, they elucidated that metaproteomics could provide a link among microbial community structure, function, physiology, interaction, ecology, and evolution. These approaches are reviewed here to help gain insights into the function of microbial community in ecosystems.
Physiological Characteristics and Angiotensin Converting Enzyme Inhibitory Activity of Lactobacillus brevis HLJ59 Isolated from Salted Shrimp
Jeon, Chun-Pyo ; Kim, Yun-Hoi ; Lee, Jung-Bok ; Jo, Min-Sub ; Shin, Kee-Sun ; Choi, Chung-Sig ; Kwon, Gi-Seok ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 9~14
In this study, lactic acid bacteria with high angiotensin converting enzyme inhibitor activity were isolated from Korean fermented food, such as kimchi and salted seafood. The strain HLJ59, isolated from salted shrimp showed the highest angiotensin converting enzyme inhibitor activity in DeMan Rogosa Sharpe broth. Optimum growth temperature of Lactobacillus brevis HLJ59 was at
. Acid treatment at pH 3.0 for 1.5 h decreased cell viability from
CFU/ml. The bile extract concentration of 0.3%, 0.5%, and 1.0% in MRS broth did not inhibit the growth of HLJ59. Isolated strain HLJ59 showed more sensibility to amikacin, gentamycin, neomycin, streptomycin, kanamycin, cefmetazole, cephalothin, ampicillin, ticarcillin, sulbactam+ampicillin, amoxicillin+clavulanic acid (AMC), tetracycline, and sulfamethoxazole+trime thoprim (SXT) as compare to other 7 different antibiotics. However, it showed more resistance to cefoxatin, ceftnaxone, penicillin, ciprofloxacin, nalidixic acid, lincomycin, and chloramphenicol.
The Infectivity of Recombinant Porcine Endogenous Retrovirus (PERV-A/C) Is Modulated by Membrane-Proximal Cytoplasmic Domain of PERV-C Envelope Tail
Kim, Sae-Ro-Mi ; Park, Sang-Min ; Lee, Kyu-Jun ; Lee, Yong-Jin ; Bae, Eun-Hye ; Park, Sung-Han ; Lim, Ji-Hyun ; Jung, Yong-Tae ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 15~20
Xenotransplantation of pig organs is complicated by the existence of polytropic replication-competent porcine endogenous retroviruses (PERV) capable of infecting human cells. Two classes of infectious human-tropic replication-competent PERVs (PERV-A and PERV-B) and one class of ecotropic PERV-C are known. The potential for recombination between ecotropic PERV-C and human-tropic PERVs adds another level of infectious risk. A recombinant PERV-A/C (PERV-A14/220) virus is 500-fold more infectious than PERV-A. Two determinants of this high infectivity was identified; one was isoleucine-to-valine substitution at position 140 in RBD (receptor binding domain), and the other lies within the PRR (proline rich region) of the envelope protein. To examine whether the effects of the cytoplasmic tail of the PERV-C Env on fusogenesity also influences infectivity, we constructed a pseudotype retroviral vectors containing MoMLV core protein and PERV envelopes. Pseudotyping experiments with the PERV envelope glycoproteins indicated that recombinant PERV-A/C virus is 10-fold more infectious than PERV-A by lacZ staining. This result supports the suggestion that viral transduction of PERV-A/C is enhanced by a membrane-proximal cytoplasmic amphiphilic
-helix in PERV-C Env tail.
Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis
Park, Moon-Kyoo ; Jung, Kyoung-Hwa ; Kim, Yeon-Hee ; Rhie, Gi-Eun ; Chai, Young-Gyu ; Yoon, Jang-W. ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 21~26
As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately
clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.
The Development of Treatment System for Removing the Low Concentrated Nitrogen and Phosphorus Using Phototrophic Bacteria and Media
Kim, Sun-Jung ; Lee, Sang-Seob ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 27~32
We used phototrophic bacteria to remove low concentrated organic materials (CODCr), nitrogen and phosphorus. We applied
4.0 mg/L, and
1.0 mg/L (C:N:P=100:10:1) in the batch test, and the removal efficiencies were shown as follow:
79.7%. The aerobic process with mixed phototrophic bacteria, ceramic media, and media KSP01 showed the removal efficiencies of
, each as 72.7% and 79.2%, respectively in the lab-scale reactor. The maximum
removal efficiency reached 92.6% by adjusting pH. There were three conditions used to remove
. The highest removal efficiency was 98.5% with 10.2 L/min of aeration in 1-2 reactors, and the result of applying river-water showed the high removal efficiency of
(82.8%). Therefore, this purification system may be useful to control nitrogen and phosphorus at low concentration in field.
Plant Growth Promotion by Isolated Strain of Bacillus subtilis for Revegetation of Barren Lakeside Area
Kim, Kyung-Mi ; Song, Hong-Gyu ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 33~37
Rhizobacterial strain isolated from barren soil, Bacillus subtilis RFO41 exhibits a high level of phosphate solubilizing activity and produces some phytohormones. Its promoting effect on the growth of Xanthium italicum Moore, a wild plant growing at lakeside barren land and thus a good candidate plant for revegetation of barren lakeside was evaluated in the in situ test for 19 weeks at Lake Paro, Kangwon-do. Strain RFO41 could enhance the dry weight of X. italicum by 67.7%. It also increased the shoot length of X. italicum plant by 21.1% compared to that of uninoculated control. Both growth enhancements had statistical significance. However, the inoculation did not show any effect on the root growth, which might be due to the breakage of tiny root. Denaturing gradient gel electrophoresis analysis showed that the inoculated bacteria were maintained in the soils, and the indigenous bacterial community did not exhibit any significant change. This plant growth promoting capability may be utilized as an environment-friendly and low cost revegetation method, especially for the sensitive areas such as barren lakeside lands.
The Types of Extended-Spectrum
-Lactamases Isolated from Suyeong Sewage Disposal Plant, Busan Environmental Corporation
Kim, Gun-Do ; Lee, Hun-Ku ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 38~45
The study performed to identify the type of ESBL against strains which are producing extendedspectrum
-lactamases and isolated from sewage in Suyeong sewage disposal plant, Busan Environmental Corporation. By the standard activated sludge method, Suyeong sewage disposal plant purify living and lavatory sewage gathering from the northeast Busan and the facility purify total 550,000 tons of living sewage disposal a day. 14 strains were isolated by double disk synergy test and the third generation cepha-antibiotics test. Indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylasedihydrolase and sugar-fermentation tests identified as Klebsiella pneumoniae (n=4) and Escherichia coli (n=10). Plasmid-mediated transmission test against isolated 14 strains proved 11 strains transmitted resistance to recipient E. coli J53 (sodium
). 9 strains of conjugant were expressed ESBL genes transferred from parental strain but 2 conjugants did not expressed. The type of ESBL from each strain was determined by isoelectric focusing points, DNA and amino acids sequencing. The results indicated that the types of ESBL transmitted to recipient E. coli J53 were TEM-1, the parental TEM type and SHV-12 type.
Optimization of Fermentation Conditions for CoQ10 Production Using Selected Bacterial Strains
Jeong, Keun-Il ; Kang, Won-Hwa ; Lee, Jung-Ah ; Shin, Dong-Ha ; Bae, Kyung-Sook ; Park, Ho-Young ; Park, Hee-Moon ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 46~51
Coenzyme Q10 (CoQ10) is an essential lipid-soluble component of membrane-bound electron transport chains. CoQ10 is involved in several aspects of cellular metabolism and is increasingly being used in therapeutic applications for several diseases. Despite the recent accomplishments in metabolic engineering of Escherichia coli for CoQ10 production, the production levels are not yet competitive with those by fermentation or isolation. So we tested several microorganisms obtained from the KCTC of Biological Resource Center to find novel sources of strain-development for CoQ10-production. Then we selected two strains, Paracoccus denitrificans (KCTC 2530) and Asaia siamensis (KCTC 12914), and tested to optimize the CoQ10 production conditions. Among the carbon sources tested, CoQ10 production was the highest when fructose was supplied about 4% concentration. Yeast extract produced the highest CoQ10 production about 2% concentration. The highest CoQ10 production was obtained at pH 6.0 for P. denitrificans and pH 8.0 for A. siamensis. And two strains showed the highest CoQ10 production at
, but the highest DCW was obtained at
. In the fed-batch culture, P. denitrificans yielded
mg and A. siamensis yielded
mg of final CoQ10 production.
Efficacy of Mixture of Lactic Acid Bacteria (LAB) and Bifidobacteria Supplement in the Management of Constipation; Demonstration of Functionality in Animal and Clinical Trials
Kim, Jung-Rae ; Lee, Do-Kyung ; Baek, Eun-Hye ; An, Hyang-Mi ; Yang, Hwan-Jin ; Kim, Mi-Jin ; Choi, Kyung-Soon ; Yun, Mi-Eun ; Jung, Yi-Jung ; Oh, Pok-Ja ; Chung, Myung-Jun ; Ha, Nam-Joo ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 52~62
The aim of this study was to evaluated the efficacy of mixture of Lactic Acid Bacteria (LAB) and bifidobacteria supplement, which are contained with Lactobacillus acidophilus, Bifidobacterium longum SPM1205, and Pediococcus pentosaceus for the management of constipation in animal model and clinical trials. 5 ICR mice and 4 female constipation subjects were orally taken mixture of LAB and bifidobacteria for 2 weeks. We investigated the number of fecal LAB and harmful enzymes activities before and after mixture of LAB and bifidobacteria application. As a result, fecal LAB count was increased and harmful enzymes activities of intestinal microflora were generally decreased after mixture of LAB and bifidobacteria application. Also, 61 female subjects were randomly assigned to receive either mixture of LAB and bifidobacteria or lactose and were taken three times a day for 2 weeks. Then, we analyzed mixture of LAB and bifidobacteria effect through the questionnaires. Daily consumption of this mixture of LAB and bifidobacteria improved the constipation in constipation group (56.3%) compared with lactose application group (26.7%). Furthermore, after mixture of LAB and bifidobacteria treatment, frequency of hard stool decreased from 0.22 to 0.03. These results indicated that mixture of LAB and bifidobacteria application is effective to improve the constipation.
Isolation of Pseudoxanthomonas sp. W12 and WD32 Producing Extracellular Protease
Cho, Woon-Dong ; Lee, Je-Kwan ; Lim, Chae-Sung ; Park, A-Rum ; Oh, Yong-Sik ; Roh, Dong-Hyun ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 63~67
Proteases catalyze hydrolytic cleavage of a peptide bond between amino acids and occupy pivotal positions in application in physiological and commercial fields. During the screening for novel bacteria producing extracellular protease, two bacterial strains, WD12 and WD32, were isolated from rotten trees and they made clear zone on LB plates supplemented with 1% skim milk. The similarities of 16S rRNA gene sequence of either WD12 or WD32 to GenBank database showed the highest to Pseuoxanthomonas mexicana as 97.8 and 99.8%, respectively. Phylogenetic analysis showed that both isolated was located within the cluster comprising P. mexicana and P. japonesis. WD12 and WD32 were catalase- and oxidase-positive, Gram-negative rod strains. In case of WD12, it could assimilate malate, but could not assimilate D-mannose, which were different characteristics from P. mexicana. Both Pseuoxanthomonas sp. WD12 and WD32 optimally produced extracellular protease at
, and maximal activity showed as 656 unit/ml and 267 unit/ml, respectively.
Characterization of Bacillus licheniformis B1
-1,4-Glucanase Overproduced in Escherichia coli
Song, Hye-Jung ; Kim, Hwang-Yeon ; Hwang, Jae-Sung ; Kim, Han-Bok ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 68~72
-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the
-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of
. The enzyme still had 76% maximal activity at
. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.
Screening for Cold-Active Protease-Producing Bacteria from the Culture Collection of Polar Microorganisms and Characterization of Proteolytic Activities
Kim, Doc-Kyu ; Park, Ha-Ju ; Lee, Yung-Mi ; Hong, Soon-Gyu ; Lee, Hong-Kum ; Yim, Joung-Han ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 73~79
The Korea Polar Research Institute (KOPRI) has assembled a culture collection of cold-adapted bacterial strains from both the Arctic and Antarctic. To identify excellent protease-producers among the proteolytic bacterial collection (874 strains), 78 strains were selected in advance according to their relative activities and were subsequently re-examined for their extracellular protease activity on
ZoBell plates supplemented with 1% skim milk at various temperatures. This rapid and direct screening method permitted the selection of a small group of 15 cold-adapted bacterial strains, belonging to either the genus Pseudoalteromonas (13 strains) or Flavobacterium (2 strains), that showed proteolytic activities at temperatures ranging between
. The cold-active proteases from these strains were classified into four categories (serine protease, aspartic protease, cysteine protease, and metalloprotease) according to the extent of enzymatic inhibition by a class-specific protease inhibitor. Since highly active and/or cold-adapted proteases have the potential for industrial or commercial enzyme development, the protease-producing bacteria selected in this work will be studied as a valuable natural source of new proteases. Our results also highlight the relevance of the Antarctic for the isolation of protease-producing bacteria active at low temperatures.
Characterization of Barley
-Amylase Chimeric Enzymes Expressed in Pichia pastoris
Kim, Tae-Jip ; Yuk, Jeong-Bin ; Choi, Seung-Ho ; Jang, Myoung-Uoon ; Svensson, Birte ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 80~85
-amylase isozymes (AMY1 and AMY2) found in barley malt share up to 80% of amino acid sequence identity with each other, but their enzymatic properties differ remarkably. AMY1 shows the highest activity at low concentration of calcium ion, while AMY2 is highly active at high calcium concentration. Meanwhile, BASI (Barley
-Amylase/Subtilisin Inhibitor) protein specifically inhibits only AMY2. In the present study, three separate regions in AMY genes (I, II, and III) were assigned on the basis of restriction enzyme sites and four kinds of chimeric amylases have been obtained by swapping a part of regions with each other. Each chimera gene was successfully over-expressed in Pichia pastoris. From the results of enzymatic characterization, both AMY211 and AMY122 showed the mixed or intermediate type of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY221 chimera could be significantly inhibited by BASI protein. As a result, it can be proposed that some amino acid residues in the region I and II, except region III, of barley
-amylases play very important roles in calcium-dependency and interaction with BASI.
Isolation and Characterization of Keratinolytic Protein Chicken Feather-Degrading Bacteria
Kim, Se-Jong ; Cho, Chun-Hwi ; Whang, Kyung-Sook ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 86~92
Thirty-one chicken feather-degrading bacteria were isolated from wasted feather, compost and wastewater in a chicken farm. These isolates were categorized as Firmicutes (21 strains),
-proteobacteria (4 strains), Actinobacteria (4 strains), and Bacteroidetes (2 strains) by 16S rRNA gene sequence analysis. We examined the feather-degrading isolates for degradation in the 2% of chicken feather meal. The strain Chryseobacterium sp. FBF-7, Stenotrophomonas maltophilia FBS-4, and Lysinibacillus sp. FBW-3 were selected as a keratinolytic protein degrading bacteria which showed the highest feather degradation of 75-90%. The characteristics of amino acids extracted from chicken feather meal by using keratinolytic protein degrading isolates and chemical method with
were analyzed. Total amino acid content of strain Chryseobacterium sp. FBF-7 was 1,661.6
/ml, which was the highest and it was similar with chemical method. And essential amino acid content of total amino acid was thirty-seven percent (619.3
/ml) and 596.9
/ml for keratinolytic protein degrading isolates and chemical method, respectively. The major amino acids were valine, glutamic acid, aspartic acid, glycine, and proline by the strain Chryseobacterium sp. FBF-7 and especially, higher contents of aspartic acid, threonine, serine, cysteine, and tyrosine were detected compared with chemical method.
Dye Removal by Phlebia tremellosa and Lignin Degrading Enzyme Transformants
Kum, Hyun-Woo ; Ryu, Sun-Hwa ; Lee, Sung-Suk ; Choi, Hyoung-T. ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 93~95
White rot fungi which have lignin degrading enzymes show high degrading activity to diverse recalcitrant compounds such as polycyclic aromatic compounds, dyes, explosives and endocrine disrupting chemicals. We have examined decolorizing activity of dyes by Phlebia tremellosa and two transformants which had genetically transformed using laccase or manganese peroxidase (MnP) gene. In case of methyl green, wild type strain showed 50% decolorization while laccase transformant (TF2-1) and MnP transformant (T5) showed more than 90% decolorization on day 3. Remazol brilliant blue R(RBBR) was decolorized up to 85% by two transformants while the wild type showed 67% decolorization on day 3. Transformants TF2-1 and T5 both showed increased laccase and MnP activity respectively during the whole growing phase.
Comparison of Oligosaccharyltransferase Assay Methods Using a Fluorescent Peptide
Kim, Seong-Hun ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 96~103
Oligosaccharyltransferase (OTase) catalyzes the transfer of a lipid-linked oligosaccharide (LLO) to the nascent polypeptide. Most eukaryotes have an OTase composed of a multisubunit protein complex. However, the kinetoplastid Leishmania major and the bacterium Campylobacter jejuni have only a single subunit for OTase activity, Stt3p and PglB, respectively. In this study, a new in vitro assay for OTase was developed by using a fluorescent peptide containing N-glycosylation sequon, Asn-Xaa-Thr/Ser, where Xaa can be any amino acid residue except Pro. L. major Stt3p and C. jejuni PglB as a model OTase enzyme demonstrated the formation of glycopeptides from a fluorescent peptide through OTase activities. For separation and measurement of the glycopeptides produced by the OTases, Tricine-SDS-PAGE, a lectin column and fluorospectrophotometer, and HPLC were applied. Comparison of these assay methods for analyzing a fluorescent glycopeptide showed HPLC analysis is the best method for separation of glycopeptides and nonglycosylated peptides as well as for quantify the peptides than other methods.
Purification and Fluorometric Analysis of Leucine-Responsive Regulatory Protein from Escherichia coli
Lee, Chan-Yong ; Kim, Sung-Chul ; Seo, Cho-Hee ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 104~108
We describe the construction of derivatives of wild type and mutant lrp genes that encode 6XHis-tag Lrps. These derivatives of wild type and mutant Lrp could be useful for in vitro studies including Lrp conformational changes. We show that 6XHis-tag Lrp wild type and 6XHis-tag Lrp R145W bind with similar patterns in vitro to 21 bp duplex DNA containing the consensus sequences of Lrp sites of upstream of the ilvIH operon. In addition, we report here the 6XHis-tag Lrp R145W is useful to investigate the conformational changes of Lrp in solution by using its own intrinsic fluorescence characteristics.
Application of a Modified Sublimation Method to Screen for PAH-Degrading Microorganisms
Kwon, Tae-Hyung ; Kim, Jun-Tae ; Kim, Jong-Shik ;
The Korean Journal of Microbiology, volume 46, issue 1, 2010, Pages 109~111
Recent studies have described various microorganisms that can degrade PAH, however, there are currently limited methods available to screen for PAH-degrading microorganisms. To screen for PAH-degrading microorganisms, a sublimation method (Alley, Jeremy F. and Lewis R. Brown. 2000. Appl. Environ. Microbiol. 66, 439-442) was modified to produce a simple screening system. In our results, there were several bacterial species capable of pyrene degradation including genera, Coryenbacterium, Gordonia, Rhodococcus, and Streptomyces, which have been screened from 350 bacterial isolates of commercial gasoline and oil-spilled sediment by the sublimation method. The main advantage of this method is that it (i) safely deposits an even, thin and visible layer of PAH onto the agar surface without the use of solvents and (ii) the quantity of PAH sublimed onto the agar can be easily controlled. Overall, this sublimation method may be an effective and simple technique to screen for PAH-degrading microorganisms.