Go to the main menu
Skip to content
Go to bottom
REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Journal of Microbiology and Biotechnology
Journal Basic Information
Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
Editor in Chief :
Volume & Issues
Volume 19, Issue 12 - Dec 2009
Volume 19, Issue 11 - Nov 2009
Volume 19, Issue 10 - Oct 2009
Volume 19, Issue 9 - Sep 2009
Volume 19, Issue 8 - Aug 2009
Volume 19, Issue 7 - Jul 2009
Volume 19, Issue 6 - Jun 2009
Volume 19, Issue 5 - May 2009
Volume 19, Issue 4 - Apr 2009
Volume 19, Issue 3 - Mar 2009
Volume 19, Issue 2 - Feb 2009
Volume 19, Issue 1 - Jan 2009
Selecting the target year
Distribution Patterns of the Members of Phylum Acidobacteria in Global Soil Samples
Lee, Sang-Hoon ; Cho, Jae-Chang ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1281~1287
DOI : 10.4014/jmb.0904.04017
The distribution pattern of the phylum Acidobacteria, a previously uncultured bacterial group, was investigated by molecular ecological analyses of global soil samples collected from pristine ecosystems across five continents. Acidobacterial 16S rDNAs were observed in almost all soil samples, and members of acidobacterial primer group A were detected in all samples that harbored the phylum Acidobacteria. Other primer groups, Y, G, and O, showed limited distribution patterns. We further divided the primer groups into acidobacterial subdivisions (class-level). Subdivisional distribution patterns were determined by comparing the observed T-RFs with theoretical T-RFs predicted by in silico digestion of acidobacterial 16S rDNAs. Consistent with the PCR results obtained with subgroup-specific primers, T-RFLP analyses showed that acidobacterial subdivision 1 belonging to primer group A was present in the majority of the soil samples. This study revealed that the phylum Acidobacteria could be globally distributed. At the subdivisional level, acidobacterial subdivision 1 might be the most widely distributed group in this phylum, indicating that members of subdivision 1 might be adapted to various soil environments, and members belonging to other subdivisions might be restricted to certain geographic regions or habitats.
Operon Required for Fruiting Body Development in Myxococcus xanthus
Kim, Do-Hee ; Chung, Jin-Woo ; Hyun, Hye-Sook ; Lee, Cha-Yul ; Lee, Kyoung ; Cho, Kyung-Yun ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1288~1294
DOI : 10.4014/jmb.0903.0112
We have used mutational analysis to identity four genes, MXAN3553, MXAN3554, MXAN3555, and MXAN3556, constituting an operon that is essential for normal fruiting body development in Myxococcus xanthus. Deletion of MXAN3553, which encoded a hypothetical protein, resulted in delayed fruiting body development. MXAN3554 was predicted to encode a metallopeptidase, and its deletion caused fruiting body formation to fail. Inactivation of MXAN3555, which encoded a putative NtrC-type response regulator, resulted in delayed aggregation and a severe reduction in sporulation. Fruiting bodies also failed to develop with the deletion of MXAN3556, another gene encoding a hypothetical protein.
Molecular Cloning and Expression of a Novel Protease-resistant GH-36
-Galactosidase from Rhizopus sp. F78 ACCC 30795
Yanan, Cao ; Wang, Yaru ; Luo, Huiying ; Shi, Pengjun ; Meng, Kun ; Zhou, Zhigang ; Zhang, Zhifang ; Yao, Bin ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1295~1300
DOI : 10.4014/jmb.0904.04003
A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant
-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an
-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at
and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of
soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.
Elucidation of Multifaceted Evolutionary Processes of Microorganisms by Comparative Genome-Based Analysis
Nguyen, Thuy Vu An ; Hong, Soon-Ho ; Lee, Sang-Yup ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1301~1305
DOI : 10.4014/jmb.0905.05017
The evolution of living organisms occurs via a combination of highly complicated processes that involve modification of various features such as appearance, metabolism and sensing systems. To understand the evolution of life, it is necessary to understand how each biological feature has been optimized in response to new environmental conditions and interrelated with other features through evolution. To accomplish this, we constructed contents-based trees for a two-component system (TCS) and metabolic network to determine how the environmental communication mechanism and the intracellular metabolism have evolved, respectively. We then conducted a comparative analysis of the two trees using ARACNE to evaluate the evolutionary and functional relationship between TCS and metabolism. The results showed that such integrated analysis can give new insight into the study of bacterial evolution.
Structural Investigation and Homology Modeling Studies of Native and Truncated Forms of
-Amylases from Sclerotinia sclerotiorum
Ben Abdelmalek, Imen ; Urdaci, Maria Camino ; Ali, Mamdouh Ben ; Denayrolles, Muriel ; Chaignepain, Stephane ; Limam, Ferid ; Bejar, Samir ; Marzouki, Mohamed Nejib ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1306~1318
DOI : 10.4014/jmb.0903.03013
The filamentous ascomycete Sclerotinia sclerotiorum is well known for its ability to produce a large variety of hydrolytic enzymes. Two
-amylases ScAmy54 and ScAmy43 predicted to play an important role in starch degradation were showed to produce specific oligosaccharides essentially maltotriose that have a considerable commercial interest. Primary structure of the two enzymes was established by N-terminal sequencing, MALDI-TOF masse spectrometry and cDNA cloning. The two proteins have the same N-terminal catalytic domain and ScAmy43 derived from ScAmy54 by truncation of 96 amino acids at the carboxyl-terminal region. Data of genomic analysis suggested that the two enzymes originated from the same
-amylase gene and that truncation of ScAmy54 to ScAmy43 occurred probably during S. sclerotiorum cultivation. The structural gene of Scamy54 consisted of 9 exons and 8 introns, containing a single 1,500-bp open reading frame encoding 499 amino acids including a signal peptide of 21 residues. ScAmy54 exhibited high amino acid homology with other liquefying fungal
-amylases essentially in the four conserved regions and in the putative catalytic triad. A 3D structure model of ScAmy54 and ScAmy43 was built using the 3-D structure of 2guy from A. niger as template. ScAmy54 is composed by three domains A, B, and C, including the well-known
barrel motif in domain A, have a typical structure of
-amylase family, whereas ScAmy43 contained only tow domains A and B is the first fungal
-amylase described until now with the smallest catalytic domain.
In Vitro Antioxidant Activity of 5-HMF Isolated from Marine Red Alga Laurencia undulata in Free Radical Mediated Oxidative Systems
Li, Yong-Xin ; Li, Yong ; Qian, Zhong-Ji ; Kim, Moon-Moo ; Kim, Se-Kwon ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1319~1327
DOI : 10.4014/jmb.0901.0004
Marine red algae of genus Laurencia are becoming the most important resources to produce unique natural metabolites with wide bioactivities. However, reports related to Laurencia undulata, an edible species used as folk herb, are rarely found to date. In this research, 5-hydroxymethyl-2-furfural (5-HMF) was isolated and characterized by nuclear magnetic resonance (NMR) from Laurencia undulata as well as other marine algae. The following characteristics of 5-HMF were systematically evaluated: its antioxidant activities, such as typical free-radicals scavenging in vitro by electron spin resonance spectrometry (ESR) and intracellular reactive oxygen species (ROS) scavenging; membrane protein oxidation; oxidative enzyme myeloperoxidase (MPO) inhibition; as well as expressions of antioxidative enzymes glutathione (GSH) and superoxide dismutase (SOD) on the gene level using the polymerase chain reaction (PCR) method. The results demonstrated that 5-HMF could be developed as a novel marine natural antioxidant or potential precursor for practical applications in the food, cosmetic, and pharmaceutical fields.
Sterols Isolated from Nuruk (Rhizopus oryzae KSD-815) Inhibit the Migration of Cancer Cells
Lee, Dae-Young ; Lee, Sang-Jin ; Kwak, Ho-Young ; Jung, La-Koon ; Heo, Ji-Eun ; Hong, Sung-Youl ; Kim, Gye-Won ; Baek, Nam-In ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1328~1332
DOI : 10.4014/jmb.0902.0072
An activity-guided fractionation method was used to isolate anticancer components from Nuruk (Rhizopus oryzae KSD-815:KSD-815). Dried powder of KSD-815 was extracted with 80% methanol and partitioned successively using n-hexane, ethyl acetate, n-butanol, and water. The n-hexane and n-butanol fractions showed a strong antimigratory effect on human cancer cells. Both of these fractions were subjected to separation and purification procedures using silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies to afford four purified compounds. These were identified as ergosterol peroxide (1), stigmast-5-en-
-diol (2), ergosta-7,22-dien-
-tetraol (3), and daucosterol (4), respectively, by spectroscopic methods such as nuclear magnetic resonance spectrometry, mass spectrometry, and infrared spectroscopy, and comparison with those in the literature. Compounds 1-4 were isolated from KSD-815 for the first time. Compounds 1 and 4 inhibited the migration of MDA-MB-231 cells at concentrations lower than
Identification of Carotenoids from Green Alga Haematococcus pluvialis by HPLC and LC-MS (APCI) and Their Antioxidant Properties
Ranga, Rao ; Sarada, A.R. ; Baskaran, V. ; Ravishankar, G.A. ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1333~1341
DOI : 10.4014/jmb.0905.03007
Haematococcus pluvial is, a green alga, accumulates astaxanthin (3,3'-dihydroxy-
'-carotene-4,4'-dione) upto 2-3% on a dry weight basis. In the present study, identification of carotenoids from Haematococcus cyst cell extract by HPLC and LC-MS (APCI) and their antioxidant properties were evaluated in in vitro model systems. The extract exhibited 89% and 78% antioxidant activities in the
-carotene linoleate model and the hydroxyl radical scavenging model, at 9 ppm of total carotenoid, respectively. The extract also showed 80%, 85%, and 79% antioxidant activities against lipid peroxidation in the kidney, brain, and liver of rats. Low-density lipoprotein oxidation induced by
ions was also protected (45%, 64%, and 75%) by the extract in a dose-dependent manner with different carotenoid levels. Thiobarbituric acid reactive substances concentration in the blood, liver, and kidney of rats were also significantly (p<0.005) decreased in H. pluvialis-treated rats. The potent antioxidant activity is attributable to various carotenoids present in the extract.
Production, Purification, and Characterization of Taxol and 10-DABIII from a new Endophytic Fungus Gliocladium sp. Isolated from the Indian Yew Tree, Taxus baccata
Sreekanth, D. ; Syed, A. ; Sarkar, S. ; Sarkar, D. ; Santhakumari, B. ; Ahmad, A. ; Khan, M.I. ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1342~1347
DOI : 10.4014/jmb.0904.04041
We have isolated endophytic fungi from the Indian yew tree, Taxus baccata, and then screened for taxol production. Out of the 40 fungal cultures screened, one fungus Gliocladium sp. was found to produce taxol and 10-DABIII (10-deacetyl baccatin III). These compounds were purified by TLC and HPLC and characterized using UV-spectroscopy, ESI-MS, MS/MS, and proton NMR. One liter of Gliocladium sp. culture yielded
of taxol and
of 10-DABIII. The purified taxol from the fungus showed cytotoxicity towards cancer lines HL-60 (leukemia), A431 (epidermal carcinoma), and MCF-7 (breast cancer).
2'-Hydroxylation of Genistein Enhanced Antioxidant and Antiproliferative Activities in MCF-7 Human Breast Cancer Cells
Choi, Jung-Nam ; Kim, Doc-Kyu ; Choi, Hyung-Kyoon ; Yoo, Kyung-Mi ; Kim, Ji-Young ; Lee, Choong-Hwan ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1348~1354
DOI : 10.4014/jmb.0903.0114
Bioconversion of the isoflavonoid genistein to 2'-hydroxygenistein (2'-HG) was performed using isoflavone 2'-hydroxylase (CYP81E1) heterologously expressed in yeast. A monohydroxylated product was analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and NMR spectrometry and was identified as 2'-HG. An initial bioconversion rate of 6% was increased up to 14% under optimized conditions. After recovery, the biological activity of 2'-HG was evaluated. Bioconverted 2'-HG showed higher antioxidant activity against 1,1-diphenyl-2-picryl hydrazine (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals than did genistein. Furthermore, 2'-HG exhibited greater antiproliferative effects in MCF-7 human breast cancer cells than did genistein. These results suggest that 2'-hydroxylation of genistein enhanced its antioxidant activity and cell cytotoxicity in MCF-7 human breast cancer cells.
-Mediated Apoptotic Cell Death in Mouse Neural Progenitor Cells via Modulation of P38 and MEK Signaling Pathways
Kim, Jeong-Hwan ; Choi, Woo-Bong ; Lee, Jong-Hwan ; Jeon, Sung-Jong ; Choi, Yung-Hyun ; Kim, Byung-Woo ; Chang, Hyo-Ihl ; Nam, Soo-Wan ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1355~1363
DOI : 10.4014/jmb.0906.06003
In the present study, the neuroprotective effects of astaxanthin on
-mediated apoptotic cell death, using cultured mouse neural progenitor cells (mNPCs), were investigated. To cause apoptotic cell death, mNPCs were pretreated with astaxanthin for 8 h and followed by treatment of 0.3 mM
. Pretreatment of mNPCs with astaxanthin significantly inhibited
-mediated apoptosis and induced cell growth in a dose-dependent manner. In Western blot analysis, astaxanthin-pretreated cells showed the activation of p-Akt, p-MEK, p-ERK, and Bcl-2, and the reduction of p-P38, p-SAPK/JNK, Bax, p-GSK3b, cytochrome c, caspase-3, and PARP. Because
triggers caspases activation, this study examined whether astaxanthin can inhibit caspases activation in
-treated mNPCs. After
treatment, caspases activities were prominently increased, but astaxanthin pretreatment significantly inhibited
-mediated caspases activation. Astaxanthin pretreatment also significantly recovered the ATP production ability of
-treated cells. These findings indicate that astaxanthin inhibits
-mediated apoptotic features in mNPCs. Inhibition assays with SB203580 (
, a specific inhibitor of p38) and PD98059 (
, a specific inhibitor of MEK) clearly showed that astaxanthin can inhibit
-mediated apoptotic death via modulation of p38 and MEK signaling pathways.
Antimicrobial Activity of the Cell Organelles, Lysosomes, Isolated from Egg White
Yoon, Ji-Hee ; Park, Jae-Min ; Kim, Ki-Ju ; Kim, Yang-Hoon ; Min, Ji-Ho ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1364~1368
DOI : 10.4014/jmb.0905.05053
Lysosomes, as a cell organelle type, are safe biological control agents that may be possible replacements for chemical antimicrobial agents because they are simply isolated from egg white. In this study, it was found that the lysosomes isolated from egg white exhibited pH-dependent antimicrobial activity, with the optimal activity found at pH 6.0. The efficiency of lysosomes in inhibiting bacterial growth and activity was evaluated over a 12-h treatment period. Seven different microorganisms were used as bacterial strains, and the lysosomes showed a significant antimicrobial effect against all strains. In addition, the antimicrobial activity was maintained for 100 days, and there did not appear to be any resistance of E. coli to the lysosomal activity up to the eighth culture. However, the lysosomes did not affect the viability of mammalian cells, suggesting the biocompatibility of lysosomes. These highly effective lysosomes have a bright future in the application of novel antimicrobial sources as a cell organelle type.
Continuous Production of Succinic Acid Using an External Membrane Cell Recycle System
Kim, Moon-Il ; Kim, Nag-Jong ; Shang, Longan ; Chang, Yong-Keun ; Lee, Sang-Yup ; Chang, Ho-Nam ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1369~1373
DOI : 10.4014/jmb.0903.03034
Succinic acid was produced by continuous fermentation of Actinobacillus succinogenes sp. 130Z in an external membrane cell recycle reactor to improve viable cell concentration and productivity. Using this system, cell concentration increased to 16.4 g/l at the dilution rate
, up to 3 times higher than that of batch culture, and the volumetric productivity of succinic acid increased up to 6.63 g/l/h at the dilution rate
, 5 times higher than that of batch fermentation. However, in the continuous culture using a high dilution rate, operational problems including severe membrane fouling and contamination by lactic acid producer were observed. Another succinic acid producer, Mannheimia succiniciproducens MBEL55E, was also utilized in this system, and the cell concentration and productivity of succinic acid at the dilution rate of
were found to be above 3 and 2.3 times higher, respectively, compared with those obtained at the dilution rate of
. These observations give a deep insight into the process design for a continuous succinic acid production by microorganisms.
Optimization of Capsular Polysaccharide Production by Streptococcus pneumoniae Type 3
Jin, Sheng-De ; Kim, Young-Min ; Kang, Hee-Kyoung ; Jung, Seung-Jin ; Kim, Do-Man ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1374~1378
DOI : 10.4014/jmb.0903.03027
Response surface methodology (RSM) examining the effects of five-level-three-factors and their mutual interactions was utilized to optimize the fermentation conditions to enhance capsular polysaccharide (CPS) production of Streptococcus pneumoniae type 3. Twenty experiments conducted in an 8-l lab-scale fermentor were designed to assess fermentation pH, supplemented glucose concentration, and stirring rate. The predicted highest CPS production by the obtained optimization model equation was 256.14 mg/l at optimal conditions [pH 7.5, stirring rate 180 rpm, and supplemented glucose concentration 1% (w/v)]. The validity of the response model was confirmed by the good agreement between the predicted and experimental results. The maximum amount of CPS obtained was
Kinetic Study of Organic Acid Formations and Growth of Anaerobiospirillum succiniciproducens During Continuous Cultures
Lee, Pyung-Cheon ; Lee, Sang-Yup ; Chang, Ho-Nam ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1379~1384
DOI : 10.4014/jmb.0905.05026
Succinic acid-producing Anaerohiospirillum succinkiproducens was anaerobically grown in glucose-fed continuous cultures using glucose as a carbon source, and the metabolic flexibility of A. succiniciproducens in response to varying glucose concentrations and dilution rates was examined Both succinic acid (SA) and acetic acid (AA) formation was growth-associated, and their growth-rate-related coefficients (
) and nongrowth-rate-related coefficients (
) were slightly influenced by glucose concentrations. A high glucose concentration (38 g/l) and high growth rate (
) did not induce by-product formation.
Statistical Optimization for Improved Production of Cyclosporin A in Solid-State Fermentation
Survase, Shrikant A. ; Annapure, Uday S. ; Singhal, Rekha S. ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1385~1392
DOI : 10.4014/jmb.0901.0035
This work evaluates the effect of different amino acids on production of Cyclosporin (CyA) production in solid-state fermentation that was previously optimized for different fermentation parameters by one factor at-a-time for the maximum production of CyA by Tolypocladium inflatum MTCC557. Based on the Plackett-Burman design, glycerol, ammonium sulfate,
, and inoculum size were selected for further optimization by response surface methodology (RSM). After identifying effective nutrients, RSM was used to develop mathematical model equations, study responses, and establish the optimum concentrations of the key nutrients for higher CyA production. It was observed that supplementation of medium containing (% w/w) glycerol, 1.53; ammonium sulfate, 0.95;
, 0.18; and inoculum size 6.4 ml/5g yielded a maximum of 7,106 mg/kg as compared with 6,480 mg CyA/kg substrate using one factor at-a-time. In the second step, the effect of amino acids on the production of CyA was studied. Addition of
-leucine in combination after 20 h of fermentation resulted in maximum production of 8,166 mg/kg.
Purification and Characterization of Phocaecin PI80: An Anti-Listerial Bacteriocin Produced by Streptococcus phocae PI80 Isolated from the Gut of Peneaus indicus (Indian White Shrimp)
Satish Kumar, Ramraj ; Arul, Venkatesan ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1393~1400
DOI : 10.4014/jmb.0901.0003
A bacteriocin-producing strain PI80 was isolated from the gut of Penaeus indicus (Indian white shrimp) and identified as Streptococcus phocae PI80. The bacteriocin was purified from a culture supernatant to homogeneity as confirmed by Tricine SDS-PAGE. Reverse-phase HPLC analysis revealed a single active fraction eluted at 12.94 min, and MALDI-TOF mass spectrometry analysis showed the molecular mass to be 9.244 kDa. This molecular mass does not correspond to previously described streptococcal bacteriocins. The purified bacteriocin was named phocaecin PI80 from its producer strain, as this is the first report of bacteriocin production by Streptococcus phocae. The bacteriocin exhibited a broad spectrum of activity and inhibited important pathogens: Listeria monocytogenes, Vibrio parahaemolyticus, and V. fischeri. The antibacterial substance was also sensitive to proteolytic enzymes: trypsin, protease, pepsin, and chymotrypsin, yet insensitive to catalase, peroxidase, and diastase, confirming that the inhibition was due to a proteinaceous molecule (i.e., the bacteriocin), and not due to hydrogen peroxide or diacetyl. Phocaecin PI80 moderately tolerated heat treatment (up to
for 10 min) and resisted certain solvents (acetone, ethanol, and butanol). A massive leakage of
ions from E. coli
, L. monocytogenes, and V. parahaemolyticus was induced by phocaecin PI80, as measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICPOES). Therefore, the results of this study show that phocaecin PI80 may be a useful tool for inhibiting L. monocytogenes in seafood products that do not usually undergo adequate heat treatment, whereas the cells of Streptococcus phocae PI80 could be used to control vibriosis in shrimp farming.
Effect of Recombinant Lactobacillus Expressing Canine GM-CSF on Immune Function in Dogs
Chung, Jin-Young ; Sung, Eui-Jae ; Cho, Chun-Gyu ; Seo, Kyoung-Won ; Lee, Jong-Soo ; Bhang, Dong-Ha ; Lee, Hee-Woo ; Hwang, Cheol-Yong ; Lee, Wan-Kyu ; Youn, Hwa-Young ; Kim, Chul-Joong ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1401~1407
DOI : 10.4014/jmb.0904.04010
Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either
colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.
Influence of Growth Conditions on Plasmid DNA Production
Silva, Filomena ; Passarinha, Luis ; Sousa, Fani ; Queiroz, Joao A. ; Domingues, Fernanda C. ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1408~1414
DOI : 10.4014/jmb.0805.0329
The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli
. bearing the pVAXI-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and
), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dcw was achieved, proving that this strategy can improve pDNA yield even at a small scale.
Occurrence of Thermophilic Campylobacter spp. Contamination on Vegetable Farms in Malaysia
Chai, L.C. ; Ghazali, F.M. ; Bakar, F.A. ; Lee, H.Y. ; Suhaimi, L.R.A. ; Talib, S.A. ; Nakaguchi, Y. ; Nishibuchi, M. ; Radu, S. ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1415~1420
DOI : 10.4014/jmb.0901.0002
The aim of the present study was to examine the prevalence of thermophilic Campylobacter spp. (Campylobacter jejuni and Campylobacter coli) in soil, poultry manure, irrigation water, and freshly harvested vegetables from vegetable farms in Malaysia. C. jejuni was detected in 30.4% and 2.7% of the soil samples, 57.1 % and 0% of the manure samples, and 18.8% and 3% of the vegetable samples from farm A and farm B, respectively, when using the MPN-PCR method. Campylobacter spp. was not found in any of the irrigation water samples tested. Therefore, the present results indicate that the aged manure used by farm A was more contaminated than the composted manure used by farm B. Mostly, the leafy and root vegetables were contaminated. C. coli was not detected in any of the samples tested in the current study. Both farms tested in this study were found to be contaminated by campylobacters, thereby posing a potential risk for raw vegetable consumption in Malaysia. The present results also provide baseline data on Campylobacter contamination at the farm level.
Bioprocess of Triphenylmethane Dyes Decolorization by Pleurotus ostreatus BP Under Solid-State Cultivation
Yan, Keliang ; Wang, Hongxun ; Zhang, Xiaoyu ; Yu, Hongbo ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1421~1430
DOI : 10.4014/jmb.0901.0033
With an aim to evaluate dye decolorization by white rot fungus on natural living conditions, reproducing by solid-state fermentation, the process of triphenylmethane dyes decolorization using the white rot fungus P. ostreatus BP, cultivated on rice straw solid-state medium, has been demonstrated. Three typical dyes, including malachite green, bromophenol blue, and crystal violet, were almost completely decolorized by the fungus after 9 days of incubation. During the process of dye decolorization, the activities of enzyme secreted by the fungus, and the contents of soluble components, such as phenolic compounds, protein, and sugar, changed regularly. The fungus could produce ligninolytic, cellulolytic, and hemicellulolytic enzymes and laccase was the most dominant enzyme in solid-state medium. Laccase, laccase isoenzyme, and the laccase mediator could explain the decolorization of malachite green, bromophenol blue, and crystal violet by the fungus in solid-state medium, respectively. It is worth noting that the presence of the water-soluble phenolic compounds could stimulate the growth of fungus, enhance the production of laccase, and accelerate dye decolorization.
Isolation and Characterization of a Plant Growth-Promoting Rhizobacterium, Serratia sp. SY5
Koo, So-Yeon ; Cho, Kyung-Suk ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1431~1438
DOI : 10.4014/jmb.0904.04014
The role of plant growth-promoting rhizobacteria (PGPR) in the phytoremediation of heavy-metal-contaminated soils is important in overcoming its limitations for field application. A plant growth-promoting rhizobacterium, Serratia sp. SY5, was isolated from the rhizoplane of barnyard grass (Echinochloa crus-galli) grown in petroleum and heavy-metal-contaminated soil. This isolate has shown capacities for indole acetic acid production and siderophores synthesis. Compared with a non-inoculated control, the radicular root growth of Zea mays seedlings inoculated with SY5 can be increased by 27- or 15.4-fold in the presence of 15 mg-Cd/l or 15 mg-Cu/l, respectively. The results from hydroponic cultures showed that inoculation of Serratia sp. SY5 had a favorable influence on the initial shoot growth and biomass of Zea mays under noncontaminated conditions. However, under Cd-contaminated conditions, the inoculation of SY5 significantly increased the root biomass of Zea mays. These results indicate that Serratia sp. SY5 can serve as a promising microbial inoculant for increased plant growth in heavy-metal-contaminated soils to improve the phytoremediation efficiency.
Isolation of an Isocarbophos-Degrading Strain of Arthrobacter sp. scl-2 and Identification of the Degradation Pathway
Li, Rong ; Guo, Xinqiang ; Chen, Kai ; Zhu, Jianchun ; Li, Shunpeng ; Jiang, Jiandong ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1439~1446
DOI : 10.4014/jmb.0811.0626
Isocarbophos is a widely used organophosphorus insecticide that has caused environmental pollution in many areas. However, degradation of isocarbophos by pure cultures has not been extensively studied, and the degradation pathway has not been determined. In this paper, a highly effective isocarbophos-degrading strain, scl-2, was isolated from isocarbophos-polluted soil. The strain scl-2 was preliminarily identified as Arthrobacter sp. based on its morphological, physiological, and biochemical properties, as well as 16S rDNA analysis. The strain scl-2 could utilize isocarbophos as its sole source of carbon and phosphorus for growth. One hundred mg/l isocarbophos could be degraded to a non detectable level in 18 h by scl-2 in cell culture, and isofenphos-methyl, profenofos, and phosmet could also be degraded. During the degradation of isocarbophos, the metabolites isopropyl salicylate, salicylate, and gentisate were detected and identified based on MS/MS analysis and their retention times in HPLC. Transformation of gentisate to pyruvate and fumarate via maleylpyruvate and fumarylpyruvate was detected by assaying for the activities of gentisate 1,2-dioxygenase (GDO) and maleylpyruvate isomerase. Therefore, we have identified the degradation pathway of isocarbophos in Arthrobacter sp. scl-2 for the first time. This study highlights an important potential use of the strain scl-2 for the cleanup of environmental contamination by isocarbophos and presents a mechanism of isocarbophos metabolism.
Two Novel Duck Antibacterial Peptides, Avian
-Defensins 9 and 10, with Antimicrobial Activity
Ma, Deying ; Liao, Wenyan ; Wang, Ruiqin ; Han, Zongxi ; Liu, Shengwang ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1447~1455
DOI : 10.4014/jmb.0904.04028
Two novel avian
-defensins (AvBDs) isolated from duck liver were characterized and their homologies with other AvBDs were analyzed. They were shown to be duck AvBD9 and AvBD10. The mRNA expression of the two genes was analyzed in 17 different tissues from 1-28-day-old ducks. AvBD9 was differentially expressed in the tissues, with especially high levels of expression in liver, kidney, crop, and trachea, whereas AvBD10 was only expressed in the liver and kidney of ducks at all the ages investigated. We produced and purified GST-tagged recombinant AvBD9 and AvBDI0 by expressing the two genes in Escherichia coli. Both recombinant proteins exhibited antimicrobial activity against several bacterial strains. The results revealed that both recombinant proteins retained their antimicrobial activities against Staphylococcus aureus under a range of different temperatures (
) and pH values (pH 3-12).
Temperature-Dependency Urease Activity in Vibrio parahaemolyticus is Related to Transcriptional Activator UreR
Park, Kwon-Sam ; Lee, Soo-Jae ; Chung, Yong-Hyun ; Iida, Tetsuya ; Honda, Takeshi ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1456~1463
DOI : 10.4014/jmb.0902.0100
Vibrio parahaemolyticus possessing urease-positive property is relatively rare, but such strains consistently exhibit the TDH-related hemolysin (TRH) gene. In this study, we examined the effects of incubation temperature on urease activity expression, using the TH3996 and AQ4673 strains where the enzyme activity is known to be temperature-dependent and -independent, respectively. In the TH3996 strain,
-galactosidase activity was 4.4-fold lower after
cultivation than after
in a ureR-lacZ fusion strain, but temperature dependency was not found in ureD- or nikA-lacZ fusion strains. However, ureR-, ureD-, and nikA-lacZ fusions of the AQ4673 strain was not influenced by incubation temperature. We compared the promoter sequences of ureR between the above two strains. Intriguingly, we detected mismatches of two nucleotides between the two strains located at positions -66 and -108 upstream of the methionine initiation codon for UreR. Additionally, urease activity was not affected by culture temperature at either
by allelic introduction of the AQ4673 ureR gene into the TH3996 ureR deletion mutant. Taken together, our study demonstrates that the transcriptional factor UreR is involved in the temperature dependency of urease activity, and two nucleotides within the ureR promoter region are of particular importance for the urease activity dependency of V. parahaemolyticus.
Development of TaqMan Probe-Based Real-Time PCR Method for erm(A), erm(B), and erm(C), Rapid Detection of Macrolide-Lincosamide-Streptogramin B Resistance Genes, from Clinical Isolates
Jung, Jae-Hyuk ; Yoon, Eun-Jeong ; Choi, Eung-Chil ; Choi, Sung-Sook ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1464~1469
DOI : 10.4014/jmb.0902.0062
To achieve more accurate and rapid detection of macrolide-lincosamide-streptogramin B resistance genes, erm(A), erm(B), and erm(C), we developed a TaqMan probe-based real-time PCR (Q-PCR) method and compared it with conventional PCR (C-PCR), which is the most widely using erm gene identification method. The detection limit of Q-PCR was 5 fg of genomic DNA or 5-8 CFU of bacterial cells of Staphylococcus aureus. The utilization of Q-PCR might shorten the time to erm detection from 3-4 h to about 50 min. These data indicated that Q-PCR assay appears to be not only highly sensitive and specific, but also the most rapid diagnostic method. Therefore, the appropriate application of the Q-PCR assay will permit rapid and accurate identification of erm genes from clinical and other samples.
Direct Multiplex Reverse Transcription-Nested PCR Detection of Influenza Viruses Without RNA Purification
Song, Man-Ki ; Chang, Jun ; Hong, Yeong-Jin ; Hong, Sung-Hoi ; Kim, Suhng-Wook ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1470~1474
DOI : 10.4014/jmb.0905.05012
This paper describes the development a of direct multiplex reverse transcription-nested polymerase chain reaction (PCR) method, devised for simultaneous detection and typing of influenza viruses. This method combines the direct reverse transcription reaction without RNA purification with the enhancement of sensitivity and specificity of nested PCR. The method successfully detected three major human influenza viruses: influenza virus A subtype 1 (H1N1) and subtype 3 (H3N2), and influenza B virus (B). The minimum number of virus particles (pfu/ml) necessary for detection in spiked saliva samples was 200 (H1N1), 140 (H3N2), and 4.5 (B). The method's sensitivity and simplicity will be convenient for use in clinical laboratories for the detection and subtyping of influenza and possibly other RNA viruses.
Detection of Virulence-Associated Genes in Clinical Isolates of Bacillus anthracis by Multiplex PCR and DNA Probes
Kumar, Sanjay ; Tuteja, Urmil ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1475~1481
DOI : 10.4014/jmb.0902.0101
Anthrax is a zoonotic disease caused by Bacillus anthracis, and well recognized as a potential agent for bioterrorism. B. anthracis can be identified by detecting the virulence factors genes located on two plasmids, pXO1 and pXO2. The aim of the present study was to determine the presence of virulence genes in 27 isolates of B. anthracis isolated from clinical and environmental samples. For this purpose, multiplex PCR and DNA probes were designed to detect protective antigen (pag), edema factor (cya), lethal factor (lef), and capsule (cap) genes. Our results indicated that all the isolates contained all the above virulence genes, suggesting that the isolates were virulent. To the best our knowledge, this is the first study about the determination of virulence marker genes in clinical and environmental isolates of B. anthracis using multiplex PCR and DNA probes in India. We suggest that the above methods can be useful in specific identification of virulent B. anthracis in clinical and environmental samples.
Anti-Proliferative Effect of Polysaccharides from Salicornia herbacea on Induction of G2/M Arrest and Apoptosis in Human Colon Cancer Cells
Ryu, Deok-Seon ; Kim, Seon-Hee ; Lee, Dong-Seok ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1482~1489
DOI : 10.4014/jmb.0902.0063
In this study, we investigated the anti-proliferative effect of polysaccharides from Salicornia herbacea on HT-29 human colon cancer cells. Crude polysaccharides from S. herbacea (CS) were prepared by extraction with hot steam water, and fine polysaccharides from S. herbacea (PS) were obtained through further size exclusion chromatography. The anti-proliferative effect of CS and PS were measured using the MTS assay, apoptosis analysis, cell cycle analysis, and RT-PCR. HT-29 cells were treated with CS or PS at different dosages (0.5, 1, 2, 4 mg
) for 24 or 48 h. CS and PS inhibited proliferation and stimulated apoptosis of cells in a dose-dependent manner. Flow cytometric analysis after Annexin V-FITC and PI staining revealed that treatment with CS or PS increased total apoptotic death of cells to 24.99% or 91.59%, respectively, in comparison with the control (13.51 %). PS increased early apoptotic death substantially - up to 12 times more than the control. Treatment with CS or PS resulted in a concentration-dependent increase of the G2/M cell population of the cell cycle as determined by flow cytometry. G2/M arrest was induced significantly with the highest concentration (4 mg
) of PS. RT-PCR was performed to study the correlation between G2/M arrest and transcription of cell cycle control genes. The anti-proliferative activity of CS and PS was accompanied by inhibition of cyclin B1, and Cdc 2 mRNA. Moreover, both CS and PS induced expression of the p53 tumor suppressor gene and the Cdk inhibitor p21. These results suggest that polysaccharides from S. herbacea have anti-cancer activity in human colon cancer cells.
Neuronal Differentiation of PC12 Cells Cultured on Growth Factor-Loaded Nanoparticles Coated on PLGA Microspheres
Park, Keun-Hong ; Kim, Hye-Min ; Na, Kun ;
Journal of Microbiology and Biotechnology, volume 19, issue 11, 2009, Pages 1490~1495
DOI : 10.4014/jmb.0903.03002
The development of nanotechnology has penetrated the fields of biology and medicine, resulting in remarkable applications for tissue regeneration. In order to apply this technology to tissue engineering, we have developed nano-scaled 3D scaffolds consisting of growth factor-loaded heparin/poly(l-lysine) nanoparticles (NPs) attached to the surface of polymeric micro spheres via polyionic complex methods. Growth factor-loaded NPs were simply produced as polyelectrolyte complexes with diameters of 100-200 nm. They were then coated onto positively charged poly(lactic-co-glycolic acid) (PLGA) pretreated with polyethyleneimine to enable cell adhesion, proliferation, and stimulation of neurite outgrowth. Propidium iodide staining and
-tubulin analysis revealed that neuronal PC12 cells proliferated extensively, expressed significant amounts of b-tubulin, and showed well-structured neurite outgrowth on polymeric microspheres by stimulation with growth factors. These results suggest that cellular adhesion and biological functionality on prepared PLGA microspheres enabled terminal differentiation of neuronal cells.