• Korean Journal of Microbiology
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• v.50 no.3
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• pp.185-190
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• 2014

The aim of this study was to investigate the prevalence of quinolone resistant E. coli from raw bulk milk and to characterize the resistance determinants. In this study, the gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced from quinolone resistant E. coli isolates. Also, the presence of plasmid-mediated quinolone resistance (PMQR) and the expression of efflux pump genes based on quantitative real-time PCR (qRT-PCR) were investigated. Of the 487 coliform bacteria, 9 strains showed nalidixic acid resistance, and 6 of the 9 nalidixic acid resistant isolates were also ciprofloxacin resistant. These 9 strains had a single mutation at codon 83 (S83L) in gyrA, 2 of them had double mutations at codon 83 and 87 (S83L and D87N) in gyrA and 3 of the 9 isolates had single mutations at codon 80 (S80I) in parC. None of the 9 isolates harbored PMQR determinants. Compared with wild-type E. coli ATCC 25922, an over-expression of the acrB gene (2.15-5.74 fold), encoding the pump component of the AcrAB-TolC efflux pump was observed in 4 of 6 ciprofloxacin resistant isolates. This study identified the quinolone resistance mechanism of E. coli isolated from raw milk samples in Gyeonggi-do.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.15-22
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• 1998

The dnrF gene, responsible for conversion of aklavinone to $\varepsilon$-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin gene cluster of Streptomyces peucetius subsp. caesius ATCC 27952, close to drrAB, one of the anthracycline resistance genes. To characterize the enzymatic properties of the aklavinone 11-hydroxylase, the dnrF gene was overexpressed in Escherchia coli. The pET-22(+) plasmid which has the T7 promoter under the control of lacUV5 gene was used for the overexpression of the dnrF gene, and the recombinant plasmid pET213 that contains the dnrF gene linked to the T7 promoter of pET-22b(+) was introduced into the E. coli BL2l. When the expression of the dnrF gene was induced by IPTG at the final concentration of 1 mM, the induced protein could be detected in SDS-PAGE only in insoluble precipitate. The insoluble protein was electroeluted from the gel and used for the preparation of antiserum in mice. Various culture conditions were tested to maximize the expression of the aklavinone 11-hydroxylase in soluble form. The enzymatic activity was checked by the bioconversion experiment, and the protein was confirmed by the SDS-PAGE and the Western blot analysis. From the analysis of the data, it was concluded that the culture induced with IPTG at the final concentration of 0.02 mM at 37$^{\circ}C$ yielded the best productivity of active form of enzyme.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.806-813
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• 2001

This study was carried out to investigate the effects of various kinds of commercial salts including Chunil, Hanju, Guwoon and Bamboo salts on the growth of microorganisms involved in kimchi fermentation. Among various microorganisms related to the kimchi fermentation, the growth of Leuconostoc mesenteroides, Lactobacillus plantarum, Pichia membranaefaciens and E. coli was examined. Based on the conditions of kimchi fermentation, 3% and 5% concentration of each salt were studied. Also, the temperatures at 1$0^{\circ}C$, 18$^{\circ}C$ and 37$^{\circ}C$ of the cultural condition were examined. The growth of Leuconostoc mesenteroides was inhibited depending on the reduction of cultural temperature and increase of concentration of salts. There was no considerable difference on the growth of Leu. mesenteroides in the different the kind of salts. However, the growth of Lactobacillus plantarum was strongly inhibited by the 5% concentration of Bamboo salt during incubation at 18$^{\circ}C$. When Lactobacillus plantarum was cultured at 1$0^{\circ}C$, its growth was remarkably decreased regardless of the kind and concentration of salts. In the case of Pichia membranaefaciens, Bamboo salt strongly inhibited its growth at all cultural temperatures. Apparent inhibitory effect on the growth of E. coli was observed by the Bamboo salt treatment during the incubation at 18$^{\circ}C$. At the cultural temperature of 1$0^{\circ}C$, similar results obtained.

• Journal of Animal Science and Technology
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• v.59 no.2
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• pp.4.1-4.5
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• 2017

Background: Biofilms were the third-dimensional structure in the solid surface of bacteria. Bacterial biofilms were difficult to control by host defenses and antibiotic therapies. Escherichia coli (E. coli) and Salmonella were popular pathogenic bacteria that live in human and animal intestines. Essential oils are aromatic oily liquids from plant materials and well known for their antibacterial activities. Method: This study was conducted to determine effect of essential oil on anti-biological biofilm formation of E. coli and Salmonella strains in in vitro experiment. Two kinds of bacterial strains were separated from 0.2 g pig feces. Bacterial strains were distributed in 24 plates per treatment and each plates as a replication. The sample was coated with a Bacterial biofilm formation was. Result: Photographic result, Escherichia coli (E. coli) and Salmonella bacteria colony surface were thick smooth surface in control. However, colony surface in blended and single essential oil treatment has shown crack surface layer compared with colony surfaces in control. Conclusion: In conclusion, this study could confirm that essential oils have some interesting effect on anti-biofilm formation of E. coli and Salmonella strains from pig feces.

• Korean Journal of Veterinary Service
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• v.16 no.1
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• pp.41-50
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• 1993

This study was undertaken the bioserotype and drug resistance of Salmonella and Escherichia coli isolated from feces for the prevention and treatment of salmonellosis and colibacillosis in zoological animals. The results obtained from the research were as follows 1. Salmonella were isolated 19, or 4.7% from 408 samples and E. coli were isolated 12, or 40.0% from 30 diarrheal samples. 2. The biotypes in 19 Salmonella were Subspecies 1. 3. The serogroups of Salmonella isolated were 47.4% in B group, 31.6% in C, 5.3% in D and 15.8% in other, and serotype of E. coli was 100% in 0127a. 4. The antibiotic resistance of Salmonella and E. coli isolated were 13, or 68.4% and 7, or 58.3% strains, respectively 5. The multiple resistant patterns of antibiotics in Salmonella were 2drugs- and 3 drugs-resistance 30.8%, respectively, and those in E. coli were mono drug-, 2 drugs- and 7 drugs-resistance 28.6%, respectively. 6. The transferred rate of resistance to recipients (E. coli ML 1410 NA$^{r}$ ) in Salmonella was 38.5%, but that in E. coli was 71.4%.

• Korean journal of applied entomology
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• v.35 no.4
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• pp.321-325
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• 1996

To isolate a novel gene for antibacterial peptide, an inducible clone(BmInc8) was selected by differential screening strategy from Bombyx mori cDNA library prepared from lavae injected with Escherichia coli. This clone contained a cDNA insert of 564 nucleotides and encoded 59 amino acids with an apparent molecular mass of 6.3 kDa. The cDNA sequence of BmInc8 had 61.2% identity compared to that of the bactericidin from Manduca sexta and also the deduced amino acids sequences from this insert had 65% identity compared to that of the cecropin D peptide Hyalophora cecropia. The transient expression assay of this insert using prokaryotic expression vector system revealed that the expressed peptide displayed the antibacterial activity. The cDNA sequence was deposited in GenBank under the accession number U30289.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.219-225
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• 2017

An aqueous chlorine dioxide ($ClO_2$) treatment combined with highly activated calcium oxide (CaO) and mild heat was tested for inactivating naturally existing bacteria and Escherichia coli O157:H7 inoculated on fresh-cut kale. Kale samples were treated with different concentrations of $ClO_2$ (10, 30, and 50 ppm), CaO (0.01%, 0.05%, 0.1%, and 0.2%), and mild heat ($25^{\circ}C$, $45^{\circ}C$, $55^{\circ}C$, and $65^{\circ}C$) as well with combinations of 30 or 50 ppm $ClO_2$ and 0.2% CaO at $55^{\circ}C$ for 3 min. An increasing concentration of $ClO_2$ and CaO significantly reduced the microbial population compared with the control. In addition, mild heating at $55^{\circ}C$ elicited greater microbial reduction than the other temperatures. A combined treatment of 50 ppm $ClO_2$ and 0.2% CaO at $55^{\circ}C$ reduced the population of naturally existing bacteria on kale by 3.10 log colony forming units (CFU)/g, and the counts of E. coli O157:H7 were below the detection limit (1 log CFU/g). In addition, no significant differences in the Hunter color values were evident in any treatment during storage. Therefore, a combined treatment of $ClO_2$ and active CaO at $55^{\circ}C$ can be an effective sanitizing method to improve the microbiological safety of fresh-cut kale without affecting its quality.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.111-117
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• 1995

In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.520-524
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• 2004

A protein that exhibited a high similarity to a major serine transporter of Escherichia coli, SdaC, was found in Haemophilus injluenzae Rd. Also, $Na^+$-stimulated serine transport activity was detected in the cells. The sdaC of H. injluenzae was cloned and the properties of the transporter were investigated. The activity of serine transport was stimulated by $Na^+$. Uptake of $Na^+$ elicited by L-serine influx into cells was also observed, which supports the idea that L-serine is transported by a mechanism of $Na^+$serine symport. No uptake of $H^+$ elicited by L-serine influx was detected. This result was not consistent with that obtained with the homologous protein, SdaC of E. coli, which uses $H^+$as a coupling cation. The serine transport via the SdaC of H. influenzae was not inhibited by other amino acids such as threonine or D-serine like the SdaC of E. coli. Thus, the SdaC of H. influenzae is a $Na^+$-dependent L-serine specific symporter and an unusual natural mutant. The $K_m$ and the $V_{max}$, value for the serine transport in the SdaC of H. influenzae were $7.6\mu$M and 22.9 nmol/min/mg protein, respectively.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1453-1458
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• 2006

PKR-like endoplasmic reticulum (ER) kinase (PERK) is a type I transmembrane ER-resident protein containing a cytoplasmic catalytic domain with a Ser/Thr kinase activity, which is most closely related to the eukaryotic translation initiation factor-$2{\alpha}$ ($eIF2{\alpha}$) kinase PKR involved in the antiviral defense pathway by interferon. We cloned and expressed the PERK C-terminal kinase domain (cPERK) in Escherichia coli. Like PERK activation in cells under ER stress, wild-type cPERK underwent autophosphorylation when overexpressed in E. coli, whereas the cPERK(K621M) with a methionine substitution for the lysine at amino acid 621 lost the autophosphorylation activity. The activated form cPERK which was purified to near homogeneity, formed an oligomer and was able to trans-phosphorylate specifically its cellular substrate $eIF2{\alpha}$. Two-dimensional phosphoamino acids analysis revealed that phosphorylation of cPERK occurs at the Ser and Thr residues. The functionally active recombinant cPERK, and its inactive mutant should be useful for the analysis of biochemical functions of PERK and for the determination of their three-dimensional structures.