• Biomolecules & Therapeutics
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• v.12 no.3
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• pp.179-184
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• 2004

The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause for urinary tract infections. In our previous study, the Adhesin/CTXA2B was expressed as soluble recombinant chimaeric protein derived from the uropathogenic Escherichia coli adhesin genetically coupled to cholera toxin A2B (CTXA2B) subunit in Escherichia coli. Since it is very important to optimize IPTG concentration and culture temperature to maximize cell growth and productivity, These optimal culture factors were determined to increase the productivity of the expressed Adhesin/CTXA2B chimaeric protein in Escherichia coli TB1 carrying pMALfimH/ctxa2b. Our data demonstrate that optimal concentration of IPTG for increased production of chimaeric protein was 0.5 mM. Additionally, culture time was 10 hours and temperature, 37${\circ}C$.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.343-349
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• 2009

Inhibitory effect of Escherichia coli O157 adhered to Caco-2 cells by the cells of Lactobacillus plantarum K11 and the cell-free culture supernatant (CFCS) and bacteriocin prepared from this strain was investigated. As the cell counts of viable L. plantarum K11 previously adhered to Caco-2 were increased, the rate of adhesion and adherent cell counts of E. coli O157 was lower. However, because the heated L. plantarum K11 rarely have the adhesion ability to Caco-2, the adhesion rate and adherent cell counts of E. coli O157 were high. In addition, the inhibitory effects of E. coli O157 adhesion by the CFCS and bacteriocin of L. plantarum K11 were dose-dependent manner. Therefore, the inhibition of adhesion of E. coli O157 to Caco-2 may result from the antimicrobial substances such as lactic acid and bacteriocin. Moreover the inhibitory activity of adhesion by the heated bacteriocin for 30 min at 100oC was similar to activity of non-treated bacteriocin, but the activity was disappeared by treatment with protease.

• Food Science of Animal Resources
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• v.31 no.2
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• pp.158-165
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• 2011

A sensitive reverse transcriptase-PCR (RT-PCR) method to detect viable Escherichia coli O157:H7 in milk was established. The primer sets were designed based on the nucleotide sequences of the rfbE (per) and wbdN genes in the O157 antigen gene cluster of E. coli O157:H7. RT-PCR using five different primer sets yielded DNA with sizes of 655, 518, 450, and 149-bp, respectively. All five of the E. coli O157:H7 strains were detected by RT-PCR, but 11 other bacterial species were not. The sensitivity of RT-PCR was improved by adding yeast tRNA as a carrier to the crude RNA extract. The RT-PCR amplifying the 149-bp DNA fragment was the most sensitive for detecting E. coli O157:H7 and the most refractory to the bactericidal treatments. Heat treatment at $65^{\circ}C$ for 30 min was the least inhibitory of all bactericidal treatments. Treatment with RNase A strongly inhibited the RT-PCR of heated milk but not unheated milk. This study described RT-PCR methods that are specific and sensitive with a detection limit of 10 E. coli O157:H7 cells, and showed that pre-treating milk samples with RNase A improved the specificity to detect viable bacteria by RT-PCR.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.461-464
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• 2021

Escherichia coli normally colonizes the lower intestine of animals and humans, but some serotypes are foodborne pathogens. The Escherichia coli K_EC180 was isolated from swine feces that were collected from a weaner pig. In this genome announcement, E. coli K_EC180 was sequenced using PacBio RS II and Illumina NextSeq 500 platforms. The complete chromosome of E. coli K_EC180 is composed of one circular chromosome (5,017,281 bp) with 50.4% of guanine + cytosine (G + C) content, 4,935 of coding sequence (CDS), 88 of tRNA, and 22 of rRNA genes. The complete genome of E. coli K_EC180 contains the toxin genes such as shiga-like toxins (stxA and stxB).

• Korean Journal of Food Science and Technology
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• v.45 no.4
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• pp.526-529
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• 2013

This study was conducted to evaluate the disinfection effects of Staphylococcus aureus, and Escherichia coli treated with 461-nm LED and pH 5 at $15^{\circ}C$ for 10 h. S. aureus strains were decreased by about 4 log CFU/mL after 461-nm LED irradiation treatment alone for 10 h. E. coli strains were inactivated by irradiation. However, when microorganisms were subjected to a combined treatment of 461-nm LED and pH 5, both strains were inactivated by irradiation for 7 h. The highest D-value was 5.05 h for S. aureus ATCC 27664 and the lowest D-value was 1.39 h for E.coli O157: H7 ATCC 35150 (p<0.05) with 461-nm LED irradiation. For the combined treatment (461-nm LED and pH 5), the highest D-value was 1.58 h for S. aureus ATCC 19095, whereas the lowest D-value was 0.83 h for S. aureus ATCC 27664 (p<0.05). These data showed that bactericidal effects of a combination of pH 5 with 461-nm LED irradiation were enhanced compared to 461-nm LED irradiation alone.

• Journal of Veterinary Clinics
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• v.23 no.1
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• pp.41-49
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• 2006

In this study, antimicrobial activity of oriental herbal medicine extract (OHME) was tested for some organisms and the preventive effects of OHME for the colonization of Campylobacter jejuni on epithelium of small intestine were examined in Korean native broiler chickens fed a forage added 1.0% OHME. The isolated Campylobacter spp were biotyped, serotyped and the susceptiblility of isolates to antimicrobial agent were examined. The growth of Staphylococcus aureus was inhibited in 0.25% OHME. C. jejuni and C. coli were inhibited in 0.1% OHME, and Salmonella spp, Lactobacillus acidophilus and Escheichia coli 0157 were inhibited in 2.0% OHME. For the application of forage added 1.0% OHME in broiler chicken farm, the frequency of Campylobacter spp from feces, liver and spleen sample of chickens were examined during 2 weeks interval. The frequence of Campylobacter spp in feces from chickens fed assorted forage (control group) was increased from 25% in first week to 75% in seventh week. But the frequence of Campylobacter spp in feces sample from chickens 134 forage added OHME was slightly reduced from 25% in first week to 15% in seventh week. The frequence of Campylobacter spp in liver, and spleen was 13.7% and 10% respectively after seventh week in control group, but the Campylobacter spp was not isolated after fifth week in live and spleen from chickens fed forage added OHME. Isolated 56 strains of thermophilic Campyiobacter from Korean native chickens was classified as C. jejuni (76.7%), C. coli (214%) and C. laridis (1.6%). The majority of 43 isolates of C. jejuni was classified on biotype I (60.4%), II (30.2%). Most of 12 isolates of C.coli were biotype I (83.3%). Isolated 31 strains C. jejuni of showed 11 different serotype, and serotype 36 (18.6%), 17 (13.9%)were most frequent. Isolated 10 strains of C. coli showed 5 different serotypes and serotype 31 (33.3%) and 21 (25%) were relatively common. Isolated Campylobacter spp were highly susceptible to nalidixic acid, amikacin, gentamicin, colistin and chloramphenucol.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.68-72
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• 2010

The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.135-141
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• 2004

Human granulocyte colony-stimulating factor (hG-CSF) is a hematopoiesis agent that principally affects the differentiation of neutrophils in the bone marrow. At present, recombinant hG-CSF is used successfully in the treatment of chemotheraphy-induced neutropenia and its indication has been expanded to bone marrow transplantation and aplastic anemia. In this study, we have constructed rhG-CSF secretion plasmid pYRC1 in which OmpA signal sequence/hG-CSF gene was expressed under the control of the T7 promoter. rhG-CSF produced in E. coli BL21 (pYRC1) grown at $37{\circ}C$ was found in aggregates. However, 15% of the periplasmic protein was soluble rhG-CSF when the E. coli BL21 (pYRC1) was cultured at $25^{\circ}C$ for 7 h in the modified MBL medium containing 10 g/$\ell$ glucose with 10 $\mu$M IPTG induction. The production of soluble rhG-CSF in E. coli BL21 (pYRC1) using fed batch culture was also studied. In the fed batch culture system, the final yield of rhG-CSF produced from E. coli BL21 (pYRC1) was increased from 4.4 mg/$\ell$to 24 mg/$\ell$by controlling the specific growth rate from $0.43 h^{-1}$ to $0.14 h^{-1}$, and optimizing the time of induction.

• Journal of Microbiology
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• v.38 no.2
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• pp.80-87
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• 2000

The nucleotide sequence of the luxC gene coding for lux-specific fatty acyl-CoA reductase and the upstream DNA (325bp)of the structural gene from bioluminescent bacterium, Photobacterium phosphoreum, has been deternubed. An open reading frame extending for more than 20 codons in 325 bp DNA upstream of luxC was not present in both directions. The lux gene can be translated into a polypeptide of 54 kDa and the amino acid sequences of lux specific reductases of P. phosphoreum shares 80, 65, 58, and 62% identity with those of the Photobacterium leiognathi, Vibrio fischeri, Vibrio harveyi, and Xehnorhabdus luminescenens reductases, respectively. Analyses of codon usage, showing that a high frequency (2.3%) of the isoleucine codon, AUA, in the luxC gene compared to that found in Escherichia coli genes (0.2%) and its absence in the luxA and B genes, suggested that the AUA codon may play a modulator role in the expression of lux gene in E. coli. The structural genes (luxC, D, A, B, E) of the P. phosphoreum coding for luciferase (${\alpha}$,${\beta}$) and fatty acid reductase (r, s, t) polypeptides can be expressed exclusively in E. coli under the T7 phage RNA polymerase/promoter system and identificationof the [35S]methionine labelled polypeptide products. The degree of expression of lux genes in analyses of codon usage. High expression of the luxC gene could only be accomplished in a mutant E. coli 43R. Even in crude extracts, the acylated acyl-CoA reductase intermediate as well as acyl-CoA reductrase activities could be readily detected.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.5
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• pp.798-802
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• 2009

This study was performed to assess the presence of contaminated microorganisms of Escherichia coli, Clostridium perfringens, and Bacillus cereus in the 112 commercial Saengshik products. E. coli was not detected in all the samples, but C. perfringens was detected in 11 products (9.8%). The number of the bacteria was less than 100 CFU/g, which was satisfactory to KFDA microbiological requirement. B. cereus was detected less than $10^2{\sim}10^3$ CFU/g in 7 products and $10^3{\sim}10^4$CFU/g in 13 products out of 25 products. Those detected bacteria from tryptose sulphite cycloserine agar and mannitol egg yolk polymyxin agar showed the typical characteristics of Gram positive and contained lecithinase, which can decompose egg-yolks layers in the biochemical test. Therefore, much more attention must be applied to satisfy the B. cereus requirement for Saengshik products sold in the market.