• Food Science of Animal Resources
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• v.29 no.3
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• pp.310-316
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• 2009

In this study, the influence of pressure assisted mild thermal inactivation (PAMTI) on E. coli ATCC 10536 was examined at 200 MPa and temperature range of $20-50^{\circ}C$. Inactivation rate significantly increased (p<0.05) as temperature and time increased at 200 MPa. The maximum inactivation (7.91 log reduction) was obtained at $50^{\circ}C$ for 30 min under 200 MPa, which meant the complete inactivation of E. coli ATCC 10536. Inactivation kinetics were evaluated with the first order inactivation rate (k), activation energy ($E_a$), thermal death time (TDT), and z value. Kinetic parameters were significantly (p<0.05) influenced by variation temperature of PAMTI. In this study, the synergistic effect of pressure and temperature were found in the inactivation of E. coli ATCC 10536 through PAMTI.

• Korean Journal of Agricultural Science
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• v.10 no.2
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• pp.200-205
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• 1983

Transfer experiment of drug resistance and citrate utilizing ability were performed to show the relationship between drug resistance and citrate utilizing ability of 76 Citrate-Utilizing variants of Escherichia coli(Cit $^+E$. coli) isolated from cattle. The results obtained were summarized as follows; 1. Of seventy-six Cit $^+E$. coli, 36(47.4%) strains transfered their drug resistance to the E. coli ML 1410 by mating. 2. Tetracycline(TC) resistance determinants and citrate utilizing ability were transmitted in the recipient strains selected for TC, but it can be seen the segregation of streptomycin (SM) resistance determinants and citrate utilizing character in 9 recipient strains selected for SM. 3. Of seventeen TC resistance Cit $^+E$. coli, 10 strains transfered citrate utilizing character, alone. 4. The transmission of the ability to utilize citrate on Simmons citrate agar at $37^{\circ}C$, was demonstrated 52(68.4%) out of the 76 Cit $^+E$. coli.

• KSBB Journal
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• v.17 no.6
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• pp.571-576
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• 2002

Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

• KSBB Journal
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• v.23 no.5
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• pp.369-374
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• 2008

The effects of reaction temperature and the addition of various detergents on the enantioselective hyrolysis activity of the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula glutinis were investigated for the production of enantiopure styrene oxide. The recombinant E. coli harboring the EH gene from R. glutinis exhibited the enantiopreference toward (R)-styrene oxide with the maximum hydrolytic activity of $165.04{\mu}mol/min/mg$ of dry cell weight (dcw). The addition of 0.5% (w/v) Tween 20 at $10^{\circ}C$ increased the initial hydrolysis rate and enantioselectivity by 1.45-fold and 2.0-fold, respectively. Enantiopure (S)-styrene oxide was prepared with 99% ee enantiopurity and 46.0% yield (theoretical yield=50%) from 20 mM racemic styrene oxide.

• Korean Journal of Human Ecology
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• v.4 no.1
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• pp.79-92
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• 2001

E. coli transformants EC3, EC4. and EC6. harboring citron oil degrading pathway genes, were co-cultured in M9 media with citron oil as a sole carbon source at 28$^{\circ}C$. Each co-culture(EC3+EC4, EC3+EC6, EC4+EC6 and EC3+EC4+EC6) showed three to four times higher cell growth than each transformant single culture. Microbial conversion products from the co-cultures were determined by GC-MS. Linalool. 4-terpineol and ${\alpha}$-terpineol were the major common products from co-cultures. Various minor products also were detected and important in flavor characteristics of cultures.

• Korean Journal of Veterinary Service
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• v.45 no.2
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• pp.101-110
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• 2022

This study was performed to investigate infectious gastrointestinal diseases in 115 Korean cats (83 indoors and 32 outdoors) with digestive signs such as diarrhea, anorexia or abdominal distention. Detection of infectious pathogens was analyzed using real-time PCR. As a result, 85 of 115 Korean cats were detected with feline corona virus (FCoV), feline parvo virus, Group A rotavirus, Clostridium perfringens (C. perfringens), Campylobacter coli (C. coli), Campylobacter jejuni, enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli, Salmonella spp., Tritrichomonas foetus, Cyclospora cayetanensis, and Giardia lamblia. The most frequently detected pathogen was C. perfringens (52 cats, 61.2%), followed by FCoV (43 cats, 50.6%) and C. coli (16 cats, 18.8%). Also, single infection was the most common (43 cats), followed by double infection in 31 cats, triple infection in 7 cats, and quadruple infection in 4 cats. There was no significant relationship between pathogen detection and age, gender, living environment, weather, and diarrhea. However, there was a significant difference between the age group under 1 year and the age group 1~7 (P value<0.05). In this study, cats with suspected gastrointestinal infection were randomly evaluated, and other factors that could affect pathogen detection were insufficiently considered. For this reason, additional epidemiological investigations with a larger number of cats and sufficient consideration of the causes that may affect the results are needed. Nevertheless, it is thought that this study can also provide valuable information on gastrointestinal pathogens in Korean cats.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.228-234
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• 1994

Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.461-465
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• 2006

We investigated the protein productivity of the promoters for genes showing prolonged induction upon cold shock in Escherichia coli. Six low temperature inducible genes (frdA, glpB, hypB, katG, nupG, ompT) were selected based on the previously reported cDNA microarray based global transcription profiling of Escherichia coli Kl2 in response to cold shock. Their promoter regions were isolated from the genomic DNA of E. coli JM109 and expression levels induced by the promoters were examined by using green fluorescence protein (GFP) as a reporter at $15^{\circ}C$ and $37^{\circ}C$. Among the six promoters, the promoter for nupG showed the highest and prolonged expression at both temperatures and the cold inducibility of nupG promoter was not observed.

• KSBB Journal
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• v.29 no.6
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• pp.443-447
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• 2014

Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.116-124
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• 2019

Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with $GT95^{syn}$ selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3-OH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing $p2GT95^{syn}K1$, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20-OH and C3-OH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes.