• Applied Chemistry for Engineering
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• v.19 no.5
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• pp.491-496
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• 2008

The microsomal epoxide hydrolase gene (referred to as mMCEH) of Mugil cephalus was cloned by PCR, and then inserted to pColdI and pET-21b(+) vector, respectively. The recombinant E. coli possessing the recombinant plasmids exhibited the enantioperference toward (R)-styrene oxide. When enantioselective kinetic resolutions were conducted with 20 mM racemic styrene oxide, enantiopure (S)-styrene oxide was obtained with high enantiopurity more than 99% enantiomeric excess (ee) and 24.50% yield by using the recombinant E. coli harboring pET-21b(+)/mMCEH.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.47-47
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• 2010

Discharge under the water is very hard and demand considerable high voltage. But specially improved electrode can generate plasma discharge to salty water with relatively low voltage. A round shape ceramic electrode having many pinholes combined with metallic one can generate plasma. 400 volt, 10 kHz and 3 micro second pulse width were applied to repeatedly running synthetic seawater with 10 L/m velocity, containing cultivated E. coli and Bacillus. As a result, 18, 94, 99.97, 100, 100 % disinfection rates to E. coli and 17.1, 17.1, 82.9, 99.4, 99.9 % disinfection rates to Bacillus subtilis were achieved to 1, 2, 3, 4, 5 times repetitive treatment respectively. In the plasma condition, the ions and electrons are separated and new kinds of components are re-synthesized by the intensive movement of the components. Especially chlorine ions are separated and recombined to residual free chlorine like HOCl, $OCl^-$. The residual free chlorine concentrations of discharged water were 0.25, 0.88, 1.39, 1.59, 1.66 mg $Cl_2$/L after 5 times treatment respectively. Another unconfirmed radical and oxidants for example, OH, $H_2O_2$, and $O_3$ can have an effect on microorganism of course.

• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.377-383
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• 2012

Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.

• Environmental Engineering Research
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• v.12 no.2
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• pp.64-71
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• 2007

The necessity of disinfecting playground soil is an important issue, because pathogenic protozoa, bacteria, and parasite eggs remain viable for several months and can infect children. UV irradiation has been used to decontaminate water but its effectiveness on soil is unclear. We determined the efficacy of UV radiation for inactivation of an indicator bacteria, E. coli (strain ATCC 8739), on playground soil. While 99% inactivation of E. coli in the soil was readily achieved by UV radiation within 55 min at $0.4\;mW\;cm^{-2}$, complete inactivation was not achieved, even after prolonged treatment at $4\;mW\;cm^{-2}$. This was attributed to the irregular surface of the soil. A small number of E. coli escaped the UV radiation because they were situated in indentations or under small particles on the soil surface. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) confirmed that the surface characteristics of the soil is the major limiting factor in the inactivation of E. coli by UV radiation. Thus UV treatment may not be adequate for disinfecting some soils and should be carefully evaluated before being used on playground soils.

• BMB Reports
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• v.32 no.4
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• pp.363-369
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• 1999

The orf8(chlD) gene cloned from Streptomyces antibioticus T$\"{u}$99 was overexpressed using an E. coli system to confirm its biological function. Induction of the E. coli strain transformed with recombinant plasmid pRFJ 1031 containing orf8 resulted in the production of a 43,000 dalton protein. Glucose-1-phosphate thymidylyltransferase activity of the cell extract obtained from the transformed strain was 4-5 times higher than that of the control strain. The expressed protein was purified 18-fold from E. coli cell lysate using three chromatographic steps with a 17% overall recovery to near homogeneity. The N-terminal amino acid sequence of the purified protein agrees with the nucleotide sequence predicted from the orf8 gene. The SDS-PAGE estimated subunit mass of 43,000 dalton agrees well with that calculated from the amino acid composition deduced from the nucleotide sequence of the orf8 gene (43,000 Da). Also, the native enzyme has a monomeric structure with a molecular mass of 43,000 dalton. The purified protein showed glucose-1-phosphate thymidylyltransferase activity catalyzing a reversible bimolecular group transfer reaction, and was highly specific for dTTP and ${\alpha}$-D-glucose 1-phosphate as substrates in the forward reaction, and for dTDP-D-glucose and pyrophosphate in the reverse reaction.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.69-77
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• 1986

The purpose of this study was the examination for presence of K99 antigen (K99), enterotoxigenicity, 0-groups, colicin and antibiotic susceptibility among E. coil isolated from calves and cows. A total of 49(18.7%) among 262 strains, isolated from 30(26.5%) out of 113 calves and cows, possesed K99, and thirty three of 49 $K99^+$ strains produced ST. Of the strains of diarrheal calf origin which less than 15 days old, a high correlation was observed between enterotoxigenic ability and K99: 92.3% of the $K99^+$ strains produced heat stable enterotoxin(ST). In O group typing of 33 $ST^+$ strains, they belonged to O20(48.4%), O8(9.1%), O9(6.1%), O139(6.1%), O149(6.1%), O101(3.0%), O115(3.0%), except six which were untypable or autoagglutinable. Of 262 E. coil isolates, 30 strains(13.3%) produced colicin and K99 were detected in 6 strains. One hundred eighty eight strains(71.8%) of 262 E. coil isolates were resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, alone or in combination thereof. One hundred and fourteen(60.6%) out of 188 drug resistance strains carried R factor($R^+$) which were transferable to the recipients by conjugation. Sixty five $R^+$ strains(57.0%) carried thermo-sensitive R plasmid.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.759-768
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• 1994

E coli strains isolated from pigs were investigated with respect to antimicrobial resistances and prevalence of R-plasmids. Also determined were properties of R-plasmids by plasmid conjugation, curing and southern hybridization using gene probes. All of 400 E coli strains were resistant to CL and SU, and 0.3% to 96.8% of the strains were resistant to most antimicrobials such as TC, PG, AM, SM, CP, GM, EM, NM, etc, while all strains were sensitive to AK. All strains were also multiply resistant to three to twelve antimicrobials. The resistances to PG, SM, TC, AM, CP, SU and ST were transferable and supposed to be mediated by R-plasmids which were opportunistic for transposition into chromosome. Plasmids bigger in size than chromosomal DNA were considered as R-plasmids and most plasmids in small size (<4Kb) proved as cryptic plasmids or nonconjugative R-plasmids. In a strain(No 99), AM resistant property was determined from both chromosomal DNA and R-plasmid DNA which is bigger in size than chromosome.

• Journal of Life Science
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• v.21 no.12
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• pp.1710-1715
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• 2011

The bactericidal effect of nano plasma ion (NPi) which was generated by NPi was analyzed using different kinds of microorganisms, exposure times, chamber sizes, ion amounts and distance. As the result of Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Staphylococcus aureus and Bacillus subtilis were shown different in decrement. Gram-negative bacteria E. coli showed the highest percentage (96.57%) and Gram-positive bacteria B. subtilis which produced spore has the lowest percentage (57.41%). From the exposure time of NPi most of the microorganisms were extinct at an early stage. According to the size of the chamber we compared the loss of E.coli and the experiment result shown, analyzed NPi using 5 chambers $0.005m^3$ to $30m^3$ for 2 hr, that when volume of the chamber increased, saturation ion and bactericidal effect was decreased. In addition, an NPi generator installed in the $1m^3$ chamber investigated the decrement of E. coli. Saturation ion concentration increased with decrement. Finally, E. coli showed a similar reduction according to the distance from NPi generator.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.177-184
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• 2009

Three hundred lactic acid bacteria isolated from human feces were studied their probiotic characters to develop potential probiotics. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, thermal stability, antimicrobial, anticancer cell, and antiviral activity. Strain Miny-148 was selected as a potential probiotic bacterium which showed resistance to low pH, bile salts and thermal stability. On the basis of fatty acid profiles and 16S rDNA sequences analysis, the strain was identified as Lactobacillus pentosus (similarity 99.9%). The strain, L. pentosus Miny-148, showed broad antimicrobial spectrum against E. coli O157:H7, Shigella flexneri, Bacillus anthracis, Staphylococcus aureus, E. coli, Vibrio cholerae, V. vulnificus, Salmonella typhimurium, and Methicillin-resistant S. aureus (MRSA). Cell-free culture supernatant of the strain also inhibited against the growth of HT-29 colon cancer cell and transmissible gastroenterits virus.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.206-212
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• 1999

The gene orf7(oxi III) was expressed using an E. coli system in anticipation that it would encode dTDP-glucose 4,6-dehydratase which is involved in the biosynthesis of the olivose moiety of chlorothricin produced from Streptomyces antibioticus Tu99. The solubility of the expressed protein increased up to 20% under optimal induction conditions. The expressed protein was purified from the E. coli BL 21(DE3) cell lysate by a 28.5-fold purification in two chromatography steps with a 38% recovery to near homogeneity. The molecular weight and N-terminal amino acid sequence of the purified protein correlated with the predicted mass and sequence deduced from the orf7 gene. The purified protein was a homodimer with a subunit relative molecular weight of 38,000 Dalton. The expressed protein was found to exhibit dTDP-glucose 4,6-dehydratase activity and be highly specific for dTDP-glucose as a substrate. The values of K'm and V'max for dTDP-glucose were 28 $\mu$M and 295 nmol $min^{-1} (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of this enzyme.$NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.