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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 20, Issue 12 - Dec 2010
Volume 20, Issue 11 - Nov 2010
Volume 20, Issue 10 - Oct 2010
Volume 20, Issue 9 - Sep 2010
Volume 20, Issue 8 - Aug 2010
Volume 20, Issue 7 - Jul 2010
Volume 20, Issue 6 - Jun 2010
Volume 20, Issue 5 - May 2010
Volume 20, Issue 4 - Apr 2010
Volume 20, Issue 3 - Mar 2010
Volume 20, Issue 2 - Feb 2010
Volume 20, Issue 1 - Jan 2010
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Comparative Proteomic Analysis of Human Amniotic Fluid Supernatants with Down Syndrome Using Mass Spectrometry
Park, Ji-Sook ; Cha, Dong-Hyun ; Jung, Jin-Woo ; Kim, Young-Hwan ; Lee, Sook-Hwan ; Kim, Young-Jun ; Kim, Kwang-Pyo ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 959~967
DOI : 10.4014/jmb.0912.12035
Down syndrome (DS) is an abnormality of the 21st chromosome that commonly occurs in children born to older women. Thus, amniotic fluid (AF) is usually collected from such women for prenatal diagnosis. This study analyzed human AF supernatants (AFS) using a mass spectrometric (MS) approach to search for candidate biomarkers of a DS pregnancy. The AFS were collected from older pregnant women at weeks 16-18 of their gestation by amniocentesis for cytogenetic analysis. The AFS from the pregnancies carrying DS (n=4) or chromosomally normal (n=6) fetuses, as revealed by the cytogenetic analysis, were then subjected to global protein profiling based on liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Affinity chromatography was also applied prior to the LC-ESI-MS/MS to minimize the masking effect of highly abundant albumin and immunoglobulin and thereby increase the diversity of the identified proteins. As a result, at least 30 new AFS proteins were identified and 44 AFS proteins were found to be differentially expressed between the DS and normal cases, where 6 of the proteins were unique to the DS cases and 11 were unique to the chromosomally normal cases. In addition, in the DS cases, 19 AFS proteins were downregulated and 8 were upregulated to varying degrees. A Western blot analysis confirmed the LC-ESI-MS/MS data, indicating that the combined detection of apolipoprotein A-II (apoA-II) and alpha-fetoprotein (AFP) could be a potential tool for diagnosing DS cases.
Characterization of Biochemical Properties of Feline Foamy Virus Integrase
Lee, Dong-Hyun ; Hyun, U-Sok ; Kim, Ji-Ye ; Shin, Cha-Gyun ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 968~973
DOI : 10.4014/jmb.1003.03006
In order to study its biochemical properties, the integrase (IN) protein of feline foamy virus (FFV) was overexpressed in Escherichia coli, purified by two-step chromatography, (Talon column and heparin column), and characterized in biochemical aspects. For the three enzymatic reactions of the 3'-processing, strand transfer, and disintegration activities, the
ion was essentially required as a cofactor. Interestingly,
ions were found to act as effective cofactors, whereas other transition elements such as
were not. Regarding the substrate specificity, FFV IN has low substrate specificities as it cleaved in a significant level prototype foamy virus (PFV) U5 LTR substrate as well as FFV U5 LTR substrate, whereas PFV IN did not. Finally, the 3'-processing activity was observed in high concentrations of several solvents such as CHAPS, glycerol, Tween 20, and Triton X-100, which are generally used for dissolution of chemicals in inhibitor screening. Therefore, in this first report showing its biochemical properties, FFV IN is proposed to have low specificities on the use of cofactor and substrate for enzymatic reaction as compared with other retroviral INs.
Expression of Yeast Cyclophilin A (Cpr1) Provides Improved Stress Tolerance in Escherichia coli
Kim, Il-Sup ; Shin, Sun-Young ; Kim, Young-Saeng ; Kim, Hyun-Young ; Lee, Dong-Hee ; Park, Kyung-Moc ; Jin, Ingn-Yol ; Yoon, Ho-Sung ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 974~977
DOI : 10.4014/jmb.0911.11005
Cyclophilins contain the conserved activity of cis-trans peptidyl-prolyl isomerase, which is implicated in protein folding, and function as molecular chaperones. When the yeast cyclophilin A gene (cpr1) was subcloned into the prokaryotic expression vector pKM260, it was found that the expression of Cpr1 drastically increased the cell viability of E. coli BL21 when under abiotic stress conditions, as in the presence of cadmium, copper, hydrogen peroxide, heat, and SDS. Therefore, this study illustrates the importance of Cpr1 as a molecular chaperone that can improve the cellular stress responses when E. coli cells are exposed to adverse conditions, while also demonstrating its potential to increase the stability of E. coli strains utilized for the production of recombinant proteins.
Adhesive Properties, Extracellular Protein Production, and Metabolism in the Lactobacillus rhamnosus GG Strain when Grown in the Presence of Mucin
Sanchez, Borja ; Saad, Naima ; Schmitter, Jean-Marie ; Bressollier, Philippe ; Urdaci, Maria C. ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 978~984
DOI : 10.4014/jmb.0911.11007
This paper examines the probiotic bacterium Lactobacillus rhamnosus GG, and how it reacts to the presence of mucin in its extracellular milieu. Parameters studied included cell clustering, adhesion to mucin, extracellular protein production, and formation of final metabolites. L. rhamnosus GG was found to grow efficiently in the presence of glucose, N-acetylglucosamine, or mucin (partially purified or purified) as sole carbon sources. However, it was unable to grow using other mucin constituents, such as fucose or glucuronic acid. Mucin induced noticeable changes in all the parameters studied when compared with growth using glucose, including in the formation of cell clusters, which were easily disorganized with trypsin. Mucin increased adhesion of the bacterium, and modulated the production of extracellular proteins. SDS-PAGE revealed that mucin was not degraded during L. rhamnosus GG growth, suggesting that this bacterium is able to partially use the glucidic moiety of glycoprotein. This study goes some way towards developing an understanding of the metabolic and physiological changes that L. rhamnosus GG undergoes within the human gastrointestinal tract.
Biotransformation of Bioactive (-)-Mellein by a Marine Isolate of Bacterium Stappia sp.
Feng, Zhile ; Nenkep, Viviane N. ; Yun, Keum-Ja ; Zhang, Dahai ; Choi, Hong-Dae ; Kang, Jung-Sook ; Son, Byeng-Wha ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 985~987
DOI : 10.4014/jmb.1002.02012
The biological transformation of the bioactive dihydroisocoumarin, (-)-mellein, isolated from the marinederived fungus Cladosporium sp., was studied. The preparativescale culture of (-)-mellein with a marine isolate of a bacterium Stappia sp. resulted in the isolation of its oxidized metabolite, (3R,4S)-4-hydroxymellein. The stereostructure of the metabolite obtained was assigned on the basis of detailed physicochemical data analyses.
Melanin Synthesis Inhibition and Radical Scavenging Activities of Compounds Isolated from the Aerial Part of Lespedeza cyrtobotrya
Lee, Mi-Yeon ; Kim, Jin-Hee ; Choi, Jung-Nam ; Kim, Ji-Young ; Hwang, Geum-Sook ; Lee, Choong-Hwan ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 988~994
DOI : 10.4014/jmb.0905.05054
The EtOAc fraction of Lespedeza cyrtobotrya showed mushroom tyrosinase inhibitory and radical scavenging activities. Four active compounds were isolated based on Sephadex LH-20 chromatography and HPLC, and the structures were elucidated, on the basis of their LC-MS and NMR spectral data, as 2-(2,4-dihydroxyphenyl)-6-hydroxybenzofuran (1), eriodictyol-7-O-glucopyranoside (2), haginin A (3), and dalbergioidin (4), respectively. Compound (1) showed mushroom tyrosinase inhibitory activity with an
and acted as a competitive inhibitor. Furthermore,
of compound 1 reduced 50% of the melanin content on human melanoma (MNT-1) cells. The radical scavenging activities of compounds 1, 2, 3, and 4 were shown to have
values of 11.0, 24.5, 9.0, and
, respectively, in an ABTS system and
values of 42.7, 36.0, 37.7, and
, respectively, in a DPPH system. The mushroom tyrosinase inhibitory activity of the EtOAc fraction of Lespedeza cyrtobotrya was contributed by compounds 1, 3, and 4, and its radical scavenging activity was contributed by compounds 1-4.
Expression and Characterization of a Novel Deoxyribose 5-Phosphate Aldolase from Paenibacillus sp. EA001
Kim, Yong-Mo ; Choi, Nack-Shick ; Kim, Yong-Ook ; Son, Dong-Ho ; Chang, Young-Hyo ; Song, Jae-Jun ; Kim, Joong-Su ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 995~1000
DOI : 10.4014/jmb.0912.12003
A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding a protein of 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79% identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into an expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified using Ni-NTA affinity chromatography and then characterized. The optimum temperature and pH of the enzyme were
and 6.0, respectively. The specific activity for the substrate deoxyribose 5-phosphate (DR5P) was
value for DR5P was determined to be 145 mM with the
from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).
Purification and Characterization of Chitinase from a New Species Strain, Pseudomonas sp. TKU008
Wang, San-Lang ; Lin, Bo-Shyun ; Liang, Tzu-Wen ; Wang, Chuan-Lu ; Wu, Pei-Chen ; Liu, Je-Ruei ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1001~1005
DOI : 10.4014/jmb.0911.11017
The chitinase-producing strain TKU008 was isolated from soil in Taiwan, and it was identified as a new species of Pseudomonas. The culture condition suitable for production of chitinase was found to be shaking at
for 4 days in 100 ml of medium containing 1% shrimp and crab shell powder, 0.1%
, and 0.05%
(pH 7). The TKU008 chitinase was suppressed by the simultaneously existing protease, which also showed the maximum activity at the fourth day of incubation. The molecular mass of the chitinase was estimated to be 40 kDa by SDS-PAGE. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitinase were pH 7,
, pH 6-7, and <
, respectively. The chitinase was completely inhibited by
. The results of peptide mass mapping showed that 11 tryptic peptides of the chitinase were identical to the chitinase CW from Bacillus cereus (GenBank Accession No. gi 45827175) with a 32% sequence coverage.
Effect of gcl, glcB, and aceA Disruption on Glyoxylate Conversion by Pseudomonas putida JM37
Li, Xuan Zhong ; Klebensberger, Janosch ; Rosche, Bettina ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1006~1010
DOI : 10.4014/jmb.0912.12005
Pseudomonas putida JM37 metabolized glyoxylate at a specific rate of 55 g/g dry biomass/day. In order to investigate their role, three genes encoding enzymes that are potentially involved in the conversion of glyoxylate were disrupted; namely, tartronate semialdehyde synthase (gcl), malate synthase (glcB), and isocitrate lyase (aceA). Strains with transposon insertion in either of these genes were isolated from a 50,000 clone library employing a PCR-guided enrichment strategy. In addition, all three double mutants were constructed via targeted insertion of a knock-out plasmid. Neither mutation of gcl, glcB, and aceA nor any of the respective double mutations influenced glyoxylic acid conversion, indicating that P. putida JM37 may possess other enzymes and pathways for glyoxylate metabolism.
New Dioscin-Glycosidase Hydrolyzing Multi-Glycosides of Dioscin from Absidia Strain
Fu, Yao Yao ; Yu, Hong Shan ; Tang, Si Hui ; Hu, Xiang Chun ; Wang, Yuan Hao ; Liu, Bing ; Yu, Chen Xu ; Jin, Feng Xie ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1011~1017
DOI : 10.4014/jmb.0910.10039
A novel dioscin-glycosidase that specifically hydrolyzes multi-glycosides, such as 3-O-
-D-glucoside, on diosgenin was isolated from the Absidia sp.d38 strain, purified, and characterized. The molecular mass of the new dioscin-glycosidase is about 55 kDa based on SDS-PAGE. The dioscin-glycosidase gradually hydrolyzes either 3-O-
)-Rha or 3-O-
)-Rha from dioscin into 3-O-
-D-Glc-diosgenin, further rapidly hydrolyzes the other
-L-Rha from 3-O-
-D-Glc-diosgenin into the main intermediate products of 3-O-
-D-Glc-diosgenin, and subsequently hydrolyzes these intermediate products into aglycone as the final product. The enzyme also gradually hydrolyzes 3-O-
-D-glucoside from [3-O-
-D-Glc-diosgenin into diosgenin as the final product, exhibiting significant differences from previously reported glycosidases. The optimal temperature and pH for the new dioscin-glycosidase is
and 5.0, respectively. Whereas the activity of the new dioscin-glycosidase was not affected by
ions, it was significantly inhibited by
ions, and slightly affected by
Escherichia coli Arabinose Isomerase and Staphylococcus aureus Tagatose-6-Phosphate Isomerase: Which is a Better Template for Directed Evolution of Non-Natural Substrate Isomerization?
Kim, Hye-Jung ; Uhm, Tae-Guk ; Kim, Seong-Bo ; Kim, Pil ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1018~1021
DOI : 10.4014/jmb.1001.01033
Metallic and non-metallic isomerases can be used to produce commercially important monosaccharides. To determine which category of isomerase is more suitable as a template for directed evolution to improve enzymes for galactose isomerization, L-arabinose isomerase from Escherichia coli (ECAI; E.C. 184.108.40.206) and tagatose-6-phosphate isomerase from Staphylococcus aureus (SATI; E.C. 220.127.116.11) were chosen as models of a metallic and non-metallic isomerase, respectively. Random mutations were introduced into the genes encoding ECAI and SATI at the same rate, resulting in the generation of 515 mutants of each isomerase. The isomerization activity of each of the mutants toward a non-natural substrate (galactose) was then measured. With an average mutation rate of 0.2 mutations/kb, 47.5% of the mutated ECAIs showed an increase in activity compared with wild-type ECAI, and the remaining 52.5% showed a decrease in activity. Among the mutated SATIs, 58.6% showed an increase in activity, whereas 41.4% showed a decrease in activity. Mutant clones showing a significant change in relative activity were sequenced and specific increases in activity were measured. The maximum increase in activity achieved by mutation of ECAI was 130%, and that for SATI was 190%. Based on these results, the characteristics of the different isomerases are discussed in terms of their usefulness for directed evolution of non-natural substrate isomerization.
Improvement of Transformation Efficiency Through In Vitro Methylation and SacII Site Mutation of Plasmid Vector in Bifidobacterium longum MG1
Kim, Jin-Yong ; Wang, Yan ; Park, Myeong-Soo ; Ji, Geun-Eog ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1022~1026
DOI : 10.4014/jmb.1003.03014
The different cleavage patterns of pYBamy59 plasmid isolated from E. coli
and B. longum MG1 by the cell extract of B. longum MG1 suggested that the main reason for its low transformation efficiency was related to the restriction modification (R-M) system. To confirm the correlation between the R-M system and transformation efficiency, in vitro methylation and site-directed mutagenesis were performed in pYBamy59. Sequence analysis of pYBamy59 fragments digested by the cell extract of B. longum MG1 revealed that all fragments were generated by restriction of the sequence recognized by SacII endonuclease. When pYBamy59 from E. coli was methylated in vitro by CpG or GpC methyltransferase, it was protected from SacII digestion. Site-directed mutagenesis, which removed SacII sites from pYBamy59, or in vitro methylation of pYBamy59 showed 8- to 15-fold increases in the transformation efficiency over intact pYBamy59. Modification of the SacII-related R-M system in B. longum MG1 and in vitro methylation in pYBamy 59 can improve the transformation efficiency in this strain. The results showed that the R-M system is a factor to limit introduction of exogenous DNA, and in vitro modification is a convenient method to overcome the barrier of the R-M system for transformation.
Monitoring of Possible Horizontal Gene Transfer from Transgenic Potatoes to Soil Microorganisms in the Potato Fields and the Emergence of Variants in Phytophthora infestans
Kim, Sung-Eun ; Moon, Jae-Sun ; Kim, Jung-Kyu ; Yoo, Ran-Hee ; Choi, Won-Sik ; Lee, Eun-Na ; Lee, Sang-Han ; Kim, Sung-Uk ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1027~1031
DOI : 10.4014/jmb.1002.02028
To examine the possibility of horizontal gene transfer between transgenic potatoes and microorganisms in potato fields, the gene flow from transgenic potatoes containing the nucleoside diphosphate kinase 2 (NDPK2) gene to microorganisms in soils was investigated. The soil samples collected from the potato fields from March to October 2007 were examined by PCR, Southern hybridization, and AFLP fingerprinting. The NDPK2 gene from soil genomic DNAs was not detected by both PCR and Southern hybridization, indicating that gene transfer did not occur in the potato fields. In addition, no discrepancy was found in pathogenicity and noticeable changes for the appearance of variants of Phytophthora infestans in each generation when serial inoculations and the analysis of genomic DNAs by AFLP were conducted. Thus, these data suggest that transgenic potatoes do not give significant impacts on the communities of soil microorganisms and the emergence of variants, although continued research efforts may be necessary to make a decisive conclusion.
Implications of SPION and NBT Nanoparticles upon In Vitro and In Situ Biodegradation of LDPE Film
Kapri, Anil ; Zaidi, M.G.H. ; Goel, Reeta ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1032~1041
DOI : 10.4014/jmb.0912.12026
The comparative influence of two nanoparticles [viz., superparamagnetic iron oxide nanoparticles (SPION) and nanobarium titanate (NBT)] upon the in vitro and in situ low-density polyethylene (LDPE) biodegradation efficiency of a potential polymer-degrading microbial consortium was studied. Supplementation of 0.01% concentration (w/v) of the nanoparticles in minimal broth significantly increased the bacterial growth, along with early onset of the exponential phase. Under in vitro conditions,
-max shifts were quicker with nanoparticles and Fourier transform infrared spectroscopy (FTIR) illustrated significant changes in CH/
vibrations, along with introduction of hydroxyl residues in the polymer backbone. Moreover, simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis (TG-DTG-DTA) reported multiple-step decomposition of LDPE degraded in the presence of nanoparticles. These findings were supported by scanning electron micrographs (SEM), which revealed greater dissolution of the film surface in the presence of nanoparticles. Furthermore, progressive degradation of the film was greatly enhanced when it was incubated under soil conditions for 3 months with the nanoparticles. The study highlights the significance of bacteria-nanoparticle interactions, which can dramatically influence key metabolic processes like biodegradation. The authors also propose the exploration of nanoparticles to influence various other microbial processes for commercial viabilities.
Characterization of Drug-Resistant Salmonella enterica Serotype Typhimurium by Antibiograms, Plasmids, Integrons, Resistance Genes, and PFGE
Benacer, Douadi ; Thong, Kwai Lin ; Watanabe, Haruo ; Puthucheary, Savithri Devi ;
Journal of Microbiology and Biotechnology, volume 20, issue 6, 2010, Pages 1042~1052
DOI : 10.4014/jmb.0910.10028
Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes,
, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.