• Journal of Microbiology
• /
• v.42 no.1
• /
• pp.25-31
• /
• 2004

Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

• Korean Journal of Microbiology
• /
• v.47 no.3
• /
• pp.275-280
• /
• 2011

In order to evaluate the diversity and change of bacterial population during the manufacturing process of traditional soybean paste (doenjang), bacterial communities were analyzed using 16S rRNA gene-based pyrosequencing. In rice straw, the most important inoculum source for fermentation, the bacterial sequences with a relative abundance greater than 1% were assigned to four phyla, Proteobacteria (71%), Actinobacteria (20.6%), Bacteroidetes (4.2%), and Firmicutes (1.3%). Unlike bacterial community composition of rice straw, a different pattern of bacterial population in meju was observed with predominantly high abundance (99.1%) of Firmicutes. Phylum composition in young doenjang was almost same as that of meju. Major genera in young doenjang were Bacillus (81.3%), Clostridium (6.9%) and Enterococcus (6.3%) and the predominant species among bacterial population was B. amyloliquefaciens (63.6%). Abundance of the phylum Firmicutes in mature doenjang was 99.98%, which was even higher value than those in meju and young doenjang. Predominant species in mature doenjang were B. amyloliquefaciens (67.3%), B. atrophaeus (12.7%), B. methylotrophicus (6.5%), B. mojavensis (3.2%), and B. subtilis. (2.5%), which were also identified as major species of the microbial flora in meju. These results suggested that rice straw was a primary source for supplement of Bacillus species in manufacturing the traditional doenjang and that some species of Bacillus strains were mainly involved in the fermentation process of traditional doenjang.

• Korean Journal of Microbiology
• /
• v.49 no.3
• /
• pp.245-252
• /
• 2013

Pseudomonas aeruginosa and Acinetobacter baumannii are significant opportunistic pathogens in hospitals and are resistant to most antibiotics. Multidrug-resistant P. aeruginosa (MDRPA) and A. baumannii (MDRAB) cause severe human nosocomial infections and are more difficult to treat than methicillin-resistant Staphylococcus aureus (MRSA). Bifidobacteria are among of the most beneficial probiotics and have been widely studied for their antimicrobial activities. The present study explored the antimicrobial activity of Bifidobacterium sp. isolated from healthy Koreans against MDRPA and MDRAB. The antimicrobial activity of the isolates against MDRPA and MDRAB, which are resistant to ciprofloxacin, tobramycin, gentamicin, meropenem, and ceftazidime, was determined by modified broth microdilution methods using absorbance. Among all tested bifidobacteria isolates (nine B. adolescentis, three B. longum, and two B. pseudocatenulatum), the culture supernatant of B. pseudocatenulatum SPM1309 showed a strong growth inhibitory effect against MDRPA and MDRAB. No change in the turbidity of the mixture was observed during incubation, and its inhibitory effect occurred through bacteriostastic action. Moreover, the antibacterial activity was observed in the fraction with molecular weights <10 kDa of bifidobacteria culture supernatant, and the active fraction was heat-stable because it maintained its activity when heated at $70^{\circ}C$ for 10 min. The results suggest that this Bifidobacterium strain could have potential applications for alternative therapy in MDRPA and MDRAB infections.

• Korean Journal of Microbiology
• /
• v.49 no.1
• /
• pp.1-7
• /
• 2013

During meiosis, genetic recombinants are formed by homologous recombination accompanying with the programmed double-strand breaks (DSBs) and strand exchanges between homologous chromosomes. The mechanism is generated by recombination intermediates such as single-end invasions (SEIs) and double-Holliday junctions (dHJs), and followed by crossover (CO) or non-crossover (NCO) products. Our study was focused on the analysis of meiotic recombination intermediates (DSBs, SEIs, and dHJs) and final recombination products (CO and NCO). We identified these meiotic recombination intermediates using DNA physical analysis under HIS4LEU2 "hot spot" system in budding yeast, Saccharomyces cerevisiae. For DNA physical analysis, when the hot spot locus is recognized by restriction enzyme from synchronous meiotic cells, the fragmented DNA that are forming recombination intermediates can be detected and quantified through Southern hybridization analysis. Our study suggests that this system can analyze the structural change of recombination intermediates during DSB-SEI transition, double-Holiday junctions and crossover/non-crossover products in meiosis.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.13 no.9
• /
• pp.4018-4024
• /
• 2012

The purpose of this study was to application in the HACCP(Hazard Analysis Critical control) system of fried kimchi soup. The establishment of Critical limit during sterilization processing was measured by sterilization temperature, sterilization time, sensory test, storage test and pH change in storage for 30 days (May 1~30, 2012). Before sterilization, general bacteria, coliform and thermophile bacteria were detected to be $6.00{\times}10^5\;CFU/m{\ell}$, $7.50{\times}10^2\;CFU/m{\ell}$ and $2.75{\times}10^2\;CFU/m{\ell}$, respectively. In contrast, all microbial was not detected after sterilization($90{\pm}5^{\circ}C$, $22{\pm}5$ mins). The sensory test was decided as the most delicious kimchi according to $90{\pm}5^{\circ}C$, $22{\pm}5min$. In conclusion, the sterilization process of fried kimchi soup would be a great alternative to prevention, decreasing and removing of harmful microorganism, such as general bacteria, coliform and thermoduric bacteria etc. Therefore, the critical limit of sterilization temperature and time for quality control and biosafety was established at $90{\pm}5^{\circ}C$, $22{\pm}5$ mins. And it suggested that HACCP plan was necessary for monitoring method, monitoring cycle, problem solving method, education, training and record management during sterilization processing.

• Korean Journal of Environmental Agriculture
• /
• v.19 no.5
• /
• pp.401-418
• /
• 2000

1. Production of the artificial zeolite from coal ash Coal fly ash is mainly composed of several oxides including $SiO_2$ and $Al_2O_3$ derived from inorganic compounds remained after burning. As minor components, $Fe_2O_3$ and oxides of Mg, Ca, P, Ti (trace) are also contained in the ash. These components are presented as glass form resulting from fusion in the process of the combustion of coal. In other word, coal ash may refer to a kind of aluminosilicate glass that is known to easily change to zeolite-like materials by hydrothermal reaction. Lots of hot seawater is disposing near thermal power plants after cooling turbine generator periodically. Using seawater in the hydrothermal reaction caused to produce low price artificial zeolite by reduction of sodium hydroxide consumption, heating energy and water cost. As coal ash were reacted hydrothermally, peaks of quartz and mullite in the ash were weakened and disappeared, and new Na-Pl peaks were appeared strengthily. Si-O-Si bonding of the bituminous coal ash was changed to Si-O-Al (and $Fe^{3+}$) bonding by the reaction. Therefore the produced Na-Pl type zeolite had high CEC of 276.7 $cmol^+{\cdot}kg^{-1}$ and well developed molecular sieve structure with low concentration of heavy metals. 2. Utilization of the artificial zeolite in agro-environment The artificial zeolite(1g) could remove 123.5 mg of zinc, 164.7 mg copper, 184.4 mg cadmium and 350.6 mg lead in the synthetic wastewater. The removability is higher 2.8 times in zinc, 3.3 times in copper, 4.7 times in cadmium and 4.8 times in lead than natural zeolite and charcoal powder. When the heavy metals were treated at the ratio of 150 $kg{\cdot}ha^{-1}$ to the rice plant, various growth inhibition were observed; brownish discoloration and death of leaf sheath, growth inhibition in culm length, number of panicles and grains, grain ripening and rice yield. But these growth inhibition was greatly alleviated by the application of artificial zeolite, therefore, rice yield increased $1.1{\sim}3.2$ times according to the metal kind. In addition, the concentration of heavy metals in the brown rice also lowered by $27{\sim}75%$. Artificial Granular Zeolites (AGZ) was developed for the purification of wastewater. Canon exchange capacity was 126.8 $cmol^+{\cdot}kg^{-1}$. AGZ had Na-Pl peaks mainly with some minor $C_3S$ peaks in X-ray diffractogram. In addition, AGZs had various pore structure that may be adhere the suspended solid and offer microbiological niche to decompose organic pollutants. AGZ could remove ammonium, orthophosphate and heavy metals simultaneously. Mixing ratio of artificial zeolite in AGZs was related positively with removal efficiency of $NH_4\;^+$ and negatively with that of $PO_4\;^{3-}$. Root growth of rice seedling was inhibited severely in the mine wastewater because of strong acidity and high concentration of heavy metals. As AGZ(1 kg) stayed in the wastewater(100L) for 4days, water quality turned into safely for agricultural usage and rice seedlings grew normally.

• Microbiology and Biotechnology Letters
• /
• v.36 no.1
• /
• pp.6-11
• /
• 2008

To check the microbiological safety with respect to increased virulence of surviving pathogens after irradiation, in this study, the transcriptional change of vvp gene encoding metalloprotease, which is one of the typical virulence factors of Vibrio mulnificus, was monitored by real-time PCR during the course of growth cycle after reinoculation of irradiated Vibrio. When V. vulnificus was exposed to a dose of 0.5 and 1 kGy, the lag period before growth resumption of sub-cultures became longer than non-irradiated counterpart as increase of irradiation dose. In the case of non-irradiated culture, the transcription of vvp was significantly activated at 15 h after inoculation, when bacterial growth reached the stationary phase, and the highest level of pretense activity (686 U/mL) was measured at the same time. Interestingly, vvp expression of irradiated Vibrio was turned up earlier than non-irradiated Vibrio during the mid log phase of growth, whereas these rapid induction of vvp expression from irradiated cells didn't result in an increase of metalloprotease production. When Vibrio was irradiated at 0.5 and 1 kGy, the protease activities peaked at 18 h after inoculation and the levels of activities were lower 1.2- and 1.4-fold, respectively, compared to the non-irradiated counterpart. Results from this study indicate that gamma radiation is not likely to activate the virulence ability of surviving Vibrio.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.28 no.5
• /
• pp.569-576
• /
• 1995

Quality changes of canned smoked-oyster in cottonseed oil (SOCO) were investigated to determine an optimal Fo-value ( Fo) for microbiological safety and quality stability during long-term storage. The canned SOCOs were sterilized at $105^{\circ}C,\;110^{\circ}C,\;and\;115^{\circ}C$ with various Fo-values and stored at temperature range of $5^{\circ}C\;-\;50^{\circ}C$. No remarkable quality changes in pin, content of amino nitrogen, arid value and thiobarbituric acid value of the canned SOCOs sterilized at $110^{\circ}C$ with $Fo\geq5.92 min,$ were recognized at all storage temperatures. Same tendency was also recognized in the products sterilized at $105^{\circ}C\;and\;150^{\circ}C$ with $Fo=5,50\pm 0.1 min\;and\;6.50\pm0.1 min,$ while those of the canned SOCOs sterilized at $110^{\circ}C\;with\;Fo<5.92 min$ altered remarkably.

• Korean Journal of Microbiology
• /
• v.51 no.4
• /
• pp.347-354
• /
• 2015

Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

• Korean Journal of Microbiology
• /
• v.53 no.3
• /
• pp.149-155
• /
• 2017

The nitrogen phosphotransferase (PTS) system is a regulatory cascade present in most Proteobacteria, where it controls different functions. The nitrogen PTS is usually composed of $EI^{Ntr}$ (encoded by the ptsP gene), NPr (encoded by the ptsO gene), and $EIIA^{Ntr}$ (encoded by the ptsN gene). While $EIIA^{Ntr}$ plays a role in a variety of cellular processes, such as potassium homeostasis, regulation of ppGpp accumulation, nitrogen and carbon metabolisms, and regulation of ABC transporters, little information is available for a physiological role of NPr. A recent study showed that dephosphorylated NPr affects adaptation to envelope stresses in Escherichia coli. In this study, we provide another phenotype related to NPr. The ptsP mutant showed a filamentation phenotype. The filamentation phenotype of the ptsP mutant was recovered by additional deletion of the ptsO gene, but not by additional deletion of the ptsN gene, suggesting that an increased level of dephosphorylated NPr in the ptsP mutant renders cells the filamentous growth. This idea was confirmed by the fact that cells with increased levels of dephosphorylated NPr shows the filamentation phenotype. Additionally, we showed that cell size of E. coli increases with incremental dephosphorylated NPr concentrations. These results suggested that dephosphorylated NPr induces morphological change of E. coli.