• KSBB Journal
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• v.20 no.6
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• pp.431-437
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• 2005

Molecular methods were employed to investigate microorganisms in a thermophilic continuous stirred tank reactor(CSTR) used for continuous $H_2$ production. The reactor was inoculated with heat-treated anaerobic sludge and fed with a glucose-based medium. Denaturing gradient gel electrophoresis showed dynamic changes of bacterial populations in the reactor during 43 days of operation. Gas composition was constant from approximately 14 days but population shift still occurred. Populations affiliated with Fervidobactrium gondwanens and Thermoanaerobacterium thermosaccharolyticum were dominant on 21 and 41 days, respectively. Keeping pH of the medium at 5.0 could suppress methanogenic activity that was detected during initial operation period. $CH_4$ and mcrA detected in the samples obtained from the reactor or inoculum suggested the heat treatment condition employed in this study is not enough to remove methanogens in the inoculum. PCR using primer sets specific to 4 main orders of methanogens suggested that major $H_2$-consuming methanogens in the CSTR belong to the order Methanobacteriales.

• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.292-299
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• 2013

In this study, we analyzed the changes in fungal populations of red-pepper fields employing eco-friendly farming methods, such as microbial agents and crop rotation, by using polymerase chain reactions coupled with denaturing gradient gel electrophoresis (PCR-DGGE). Primer specific for fungi were used to determine the contribution of domains to the microbial community. Analysis of planted and non-planted soil samples applying PCR-DGGE technology offered evaluation of long-term patterns in fungal species richness. To evaluate the stability of DGGE patterns from different soils, comparison of planted and non-planted soil samples were compared using PCR-DGGE. The number of DNA fragments obtained from all planted soil samples by DGGE separation was far greater (14 to 15 bands) than that of the non-planted soil samples (3 to 4 bands). In addition, 14 bands were observed from crop continuation soil treated with agrochemicals and 18 bands from crop rotation soil treated with microbial agents. The PCR-DGGE analysis suggests that the use of crop rotation and microbial agents benefits the fungal community more than crop continuation using agrochemicals. These results indicate that crop rotation with microbial agents was better able to support beneficial organisms, enable more effective biological control and maintain a healthier balance of nutrients, organic matter and microorganisms.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1717-1725
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• 2013

Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.4
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• pp.294-300
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• 2013

In this study, major parameter of partial nitritation was investigated for the stable operation. In order to establish partial nitritation system, prevailing parameters such as temperature, BA (bicarbonate alkalinity) and pH were evaluated. As a result, it is inferred that appropriate bicarbonate alkalinity ratio (mg $NaHCO_3{\cdot}L^{-1}/mg$ Inf. $NH_4{^+}-N{\cdot}L^{-1}$) drives stable 50% partial nitritation at $32^{\circ}C$ and ambient temperature, respectively. Alkalinity ratio was proposed as new strategy for 50% partial nitritation without pH control in both temperature regimes. Because of the results, it was added amound of BA required only for 50% nitritation to inhibit nitratation. The effluent $NO_2{^-}-N/NH_4{^+}-N$ ratio reached almost 100% when initial bicarbonate alkalinity ratios (mg $NaHCO_3{\cdot}L^{-1}/mg$ Inf. $NH_4{^+}-N{\cdot}L^{-1}$) were 6.8 (R1) and 6.7 (R2), respectively. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) results demonstrated that AOB was the dominant nitrifying bacteria and NOB was negligible after adopting process control.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.801-808
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• 2002

Proteolytic activities of some commercial milk clotting enzymes(rennet, trypsin, pepsin, papain W-40, neutrase 1.5 and protease S) in bovine skim milk containing 0.02% $CaCl_2$ were determined by measuring DH(Degree of Hydrolysis), NPN(Non Protein Nitrogen) and by comparing patterns of SDS-PAGE(Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis). The DH of microbial enzymes(neutrase 1.5 and protease S) and trypsin in bovine skim milk were higher than those of pepsin and papain W-40. The amounts of NPN in the milk treated with trypsin and the other animal enzymes(rennet and pepsin) showed the highest and lowest degrees of proteolysis, respectively. SDS-PAGE showed that trypsin and protease S hydrolyzed $\alpha$-lactalbumin and papain W-40 hydrolyzed $\beta$-lactoglobulin slightly, while neutrase 1.5 hydrolyzed both $\alpha$-lactalbumin and $\beta$-lactoglobulin after treating for 90 min. Trypsin and protease S easily hydrolyzed ${\alpha}_s$-casein and $\beta$-casein, which were not hydrolyzed by rennet. Papain W-40 hydrolyzed $\kappa$-casein more than rennet as shown in SDS-PAGE. Based on the results of the experiments, the DH and NPN of trypsin, neutrase 1.5 and protease S were shown to be higher than those of the other enzymes. The SDS-PAGE patterns of papain W-40 and neutrase 1.5 were similar with that of rennet.

• Journal of Veterinary Clinics
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• v.21 no.1
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• pp.20-22
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• 2004

Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.

• The Journal of Internal Korean Medicine
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• v.29 no.1
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• pp.130-148
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• 2008

Objective : This study was designed to investigate the effect of the water extract of Gamisamgibopae-tang(GMSGBPT), an oriental herbal formulation, on the growth of NCI-H460 and A549 human non-small-cell lung cancer cell lines. Methods : Cytotoxicity and cell morphology were evaluated by MTT assay and inverted microscope, respectively. Apoptosis was detected using agarose gel electrophoresis and flow cytometer. The expression levels of mRNAs and proteins of target genes were determined by RT-PCR and western blot analyses, respectively Result and Conclusion : We found that exposure of A549 cells to GMSGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, but GMSGBPTdid not affect the growth of NCI-H460 cells. The anti-proliferative effect of GMSGBPT treatment in A549 cells was associated with morphological changes, formation of apoptotic bodies and DNA fragmentation, and flow cytometry analysis confirmed that GMSGBPT treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptotic cell death by GMSGBPT were connected with a up-regulation of cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) mRNA and protein in a tumor suppressor p53-independent fashion. However GMSGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1), inducible nitric oxide synthase (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 orNCI-H460 cells. Taken together, these findings partially provide novel insights into the possible molecular mechanism of the anti-cancer activity of GMSGBPT.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.155-162
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• 1988

Surface membrane proteins of virulent RH strain and tissue cyst-forming Fukaya strain of Toxoplasma gondii were analysed by SDS-polyacrylamide gel electrophoresis after LPO-catalyzed surface iodination and lectin blotting, then identified the zoite-specific antigens. Prior to the analyses, purification of RH tachyzoites from mouse peritoneal exudate and of Fukaya bradyzoites from mouse brain tissues were performed by centrifugation - on the discontinuous Percoll density-gradient. Ta- chysoites were obtained at the interface of 50U and 60% Percoll solution and brain cysts were harvested at the interfaces of 40-50% and 50-60%, then bradyzoites were obtained by treating the cysts with hypertonic solution. The LPO-catalyzed iodination detected 15 KDa and 14 KDa proteins o( brady- zoites and 30 KDa protein of tachysoites as major bands with several other minor bands. But Con A blotting revealed some bands of 200 K∼50 KDa glycoproteins of bradyzoites and 52 KDa band as major and minor bands of 33 K∼20 KDa of tachyzoites. Phytohemagglutinin did not detect any band in the two forms. EITB with anti- Fukaya antibody and anti-RH antibody revealed cross-reactivities between the two forms. Despite the cross-reactivity, anti-Fukaya antibody reacted with 15 KDa band of bradyzoites specifically and, anti-RH antibody with 52 KDa, 30 KDa, and 25 KDa bands of tachyzoites, respectively. It was identified that 15 KDa protein in bradyzoite, which was not a glycoprotein, was a major membrane protein with sufficient antigenicity, and in the case of tacky- zoite, 52 KDa surface glycoprotein (gp52) with specific antigenicity might be added to the major surface protein, p30.

• Korean Journal of Veterinary Research
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• v.29 no.1
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• pp.13-20
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• 1989

The proteins of the bovine erythrocyte membrane were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow sedimentation rate of bovine erythrocytes were investigated by treating the erythrocytes with trypsin. The erythrocyte sedimentation rates of bovine erythrocytes from Holstein and Korean native cattle were very slow compared with the human one (1/7 as slow as the human one) as reported previously. However, when human and Holstein erythrocytes were treated with trypsin (0.2 and 0.5 mg/ml) for 1 hour at ${37^{\circ}C}$, their sedimentation rates were markedly accelerated while the sedimentation rate of Korean native cattle's erythrocytes were not affected. Although the general protein profiles of the bovine erythrocyte membranes were almost similar to that of human, bovine erythrocyte membranes showed one additional protein band, called band Q in this study, which migrated electrophoretically to the mid-position between band 2 and band 3 in human erythrocyte membranes. Treatment of Holstein and human erythrocytes with trypsin caused a decrease or disapperance of the band Q from the erythrocyte membrane. Although the band Q in Korean native cattle's erythroyte membrane was decreased by trypsin treatment of the erythrocytes, the magnitude of the decrement was not so pronounced as in the case of human and Holstein erythrocytes. The glycoprotein profiles of the bovine erythrocyte membranes revealed by periodic acid-Schiff stain showed a marked difference from that of human. The PAS-1 (glycophorin) and PAS-2 (sialoglycoprotein) present in human erythrocyte membrane were almost absent from the bovine erythrocyte membranes. Instead, the bovine erythrocyte membranes showed a strong PAS-positive band near the origin of the electrophorograms, which is named as PAS-B in this study. The PAS-B band was disappered completely by the trypsin treatment of Holstein erythrocytes whereas the PAS-B band in Korean native cattle's erythrocyte membrane still remained after the trypsin treatment. The trypsin treatment of Korean native cattle's erythrocytes, however, led to the appearance of small molecular weight peptides, indicating that the high molecular weight glycoproteins were degraded by trypsin as in human and Holstein ones. These results suggest that the slow sedimentation rate of bovine erythrocytes is due in part to the presence of band Q protein fraction and PAS-B glycoprotein in the bovine erythrocytes.

• Journal of Life Science
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• v.15 no.2 s.69
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• pp.161-168
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• 2005

In this study, we did the molecular typing of 39 environmental Legionella pneumophila serogroup 1 isolates collected from 2001-2003 in Busan using the pulsed-filed gel electrophoresis (PFGE). PFGE of SfiI fragments were divided into 10 pulsotypes $(A\~J)$, corresponding to $<65\%$ similarity and a subtype within each pulsotype was characterized by $>84\%$ similarity. The major cluster was pulsotype E $(46.2\%)$, which included 18 isolates and was divided into 4 subtypes $(E1\~E4)$. PFGE of NotI fragments were divided into 8 pulsotypes $(a\~h)$, corresponding to $<60\%$ similarity and a subtype within each pulsotype was characterized by $100\%$ similarity. The major cluster was pulsotype f $(38.5\%)$, which included 15 isolates. The ATCC type strain L. pneumophila serogroup 1 was identified as a different molecular pulsotype compare to the Busan isolates. It is possible that L. pneumophila serogroup 1 isolated in Busan with specific DNA pattern is comparable with those isolation in other cities in Korea.