• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.82-85
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• 1997

The nar promoter as an inducible promoter was characterized for the process development for the gene expression and the protein production under anaerobic condition. The LB medium was selected as a main culture medium showing the enzyme activity of 18,000 units/min/g cell in the flask cultivation. The optimum concentration of nitrate was 1%. Under anaerobic conditions, the gene expression was fully induced in the presence of nitrate.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 1998.10a
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• pp.41-57
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• 1998

Five hundreds of bifidobacteria were isolated from an healthy Korean and the inhibitory effects of these isolated bacteria on harmful enzymes of human intestinal microflora were examined by cocultivation of the isolated bifidobacteria with E. coli HGU-3 or total human intestinal microflora. In comparison with the results of E. coli or intestinal microflora cultivation, Bifidobacterium breave K-110, B. breve K-111 and B. infantis K-525 effectively inhibited harmful enzymes (${\beta}-glucuronidase$ and tryptophanase) of E. coli and lowered the pH of the culture media. Also they inhibited the harmful enzymes (${\beta}-glucosidase$, ${\beta}-glucuronidase$, tryptophanase and urease) and ammonia production of intestinal microflora, and lowered pH of the culture media by increasing the number of bifidobateria on intestinal microflora. The inhibitory effect of bifidobacteria on Growth of Helicobacter pylori and Rotavirus infection were exammed. Bifidobacterium K-110 and K-111 inhibited effectively them. When these isolated bifidobacteria were administered to mice, the activities of fecal harmful enzymes were inhibited and the AC and ACF formation were suppressed. Among tested bifidobacteria, B. breve K-110 had high inhibitory effect of fecal harmful enzymes and ACF formation.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1327-1332
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• 2004

The hemA gene encoding 5-aminolevulinic acid synthase (ALAS) was cloned from Rhodobacter capsulatus, and its nucleotide sequence was determined. DNA sequencing data revealed one open reading frame coding for a protein with 401 amino acids that displayed high similarity to the amino acid sequences of other known ALASs. The hemA gene was then cloned and expressed under the control of constitutive promotor in E. coli. The recombinant E. coli strain was able to accumulate 5-aminolevulinic acid to 21 mM in the liquid medium supplemented with 45 mM glycine and 120 mM succinate. In addition, a marked effect of the pH of the culture medium on ALA production was observed, and the optimum pH for culture medium was determined to be 5.8-6.3.

• Korean Journal of Microbiology
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• v.11 no.2
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• pp.101-106
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• 1973

The microwave of 2450 MHz, generated by a household cooking oven, was evaluated for its applicability to melt various rehydrated media and to remove dissolved oxygen from tubed media for anaerobic culture. The effect on the sterilization of E. coli in selective media was also evaluated. The following results were obtained. 10 The microwave oven was useful in saving time for melting media and in eliminating heat and combustion gas from the laboratory, which were inevitable by-products in the conventional flame method. 2) Dissolved oxygen could be removed without boiling over by exposing the tubes of anaerobic culture medium after putting them in a wire basket in a beaker with water. 30 The count of E. coli during the melting of MacConkey and EMB agar were similar to those treated with open flame. The microwave treatment was not considered a possible mean to replace autoclaving even in these selective media.

• Archives of Pharmacal Research
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• v.21 no.1
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• pp.54-61
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• 1998

Five hundreds of bifidobacteria were isolated from a healthy Korean and the inhibitory effects of these isloated bacteria on harmful enzymes of human intestinal microflora were examined by cocultivation of the isolated bifidobacteria with E. coli or total human intestinal microflora. In comparison with the results of E. coli or intestinal microflora cultivation, Bifidobacterium breve K-110, B. breve K-111 and B. infantis K-525 effectively inhibited harmful enzymes ($\beta$-glucuronidase and tryptophanase) of E. coli and lowered the pH of the culture media. Also they inhibited the harmful enzymes ($\beta$-glucosidase, $\beta$-glucuronidase, tryptophanase and urease) and ammonia production of intestinal microflora, and lowered pH of the culture media by increasing lactic acid bacteria of intestinal microflora. When these isolated bifidobacteria were administered on mice, fecal harmful enzymes were also inhibited. Among tested bifidobacteria, B. breve K-110 had the highest inhibitory effect of fecal harmful enzymes.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1136-1140
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• 2008

The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'-phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.148-156
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• 2021

Single-chain antibodies against epidermal growth factor receptor variant III (EGFRvIII) are potentially promising agents for developing antibody-based cancer treatment strategies. We described in our previous study the successful expression of an anti-EGFRvIII scFv antibody in Escherichia coli. However, we could also observe the formation of insoluble aggregates in the periplasmic space, limiting the production yield of the active product. In the present study, we investigated the mechanisms by which growth conditions could affect the expression of the soluble anti-EGFRvIII scFv antibody in small-scale E. coli NiCo21(DE3) cultures, attempting to maximize production. The secreted scFv molecules were purified using Ni-NTA magnetic beads and protein characterization was performed using SDS-PAGE and western blot analyses. We used the ImageJ software for protein quantification and determined the antigen-binding activity of the scFv antibody against the EGFRvIII protein. Our results showed that the highest percentage of soluble scFv expression could be achieved under culture conditions that combined low IPTG concentration (0.1 mM), low growth temperature (18℃), and large culture dish surface area. We found moderate-yield soluble scFv production in the culture medium after lactose-mediated induction, which was also beneficial for downstream protein processing. These findings were confirmed by conducting western blot analysis, indicating that the soluble, approximately 30-kDa scFv molecule was localized in the periplasm and the extracellular space. Moreover, the antigen-binding assay confirmed the scFv affinity against the EGFRvIII antigen. In conclusion, our study reveals that low-speed protein expression is preferable to obtain more soluble anti-EGFRvIII scFv protein in an E. coli expression system.

• Journal of Dairy Science and Biotechnology
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• v.41 no.3
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• pp.126-137
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• 2023

In this study, we isolated lactic acid bacteria from Allium wakegi and examined the usability of culture supernatants obtained from these lactic acid bacteria. The antibacterial activity of the culture supernatant obtained from the isolated lactic acid bacteria against the pathogens Escherichia and Salmonella spp. was measured. The obtained lactic acid bacteria culture medium showed significant antibacterial activity against pathogenic bacteria in a dose-dependent manner. The effects of pH and heat denaturation on the observed anti-pathogenic bacterial activity was also investigated. Adjusting the culture supernatant to pH 7 resulted in loss of all antibacterial activity against pathogenic bacteria, suggesting that the antibacterial activity of the obtained culture supernatant against pathogenic bacteria is influenced by organic acids. Assessment of the heat stability of the anti-pathogenic bacterial activity revealed that heat treatment did not diminish activity. The obtained lactic acid bacteria culture medium is thus stable against heat.

• Journal of Food Hygiene and Safety
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• v.28 no.3
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• pp.234-240
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• 2013

This test was performed to evaluate the bactericidal efficacy of Fumagari OPP$^{(R)}$, fumigation disinfectant, containing 20% ortho-phenylphenol against Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium). In preliminary tests, both E. coli and S. typhimurium working culture suspension number (N value) was $4.0{\times}10^8$ CFU/mL. And all of the colony numbers on the carriers exposed the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of bacterial test suspentions by pour plate method), 0.5N2 (the number of bacterial test suspentions by filter membrane method) and 0.5N1, respectively. In addition, the mean number of bacteria recovered on the control-carriers (T value) was $3.4{\times}10^6$ CFU/mL. In the bactericidal effect of the fumigant, the reduction number of S. typhimurium and E. coli (d value) was 5.26 and 5.64 logCFU/mL, respectively. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. With the results of this study, Fumagari OPP$^{(R)}$ has an effective bactericidal activity, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with pathogenic bacteria.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.752-755
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• 2003

The antimicrobial activity of fresh garlic juice against Escherichia coli O157:H7 was investigated. When E. coli O157:H7 was cultured for 18 hr in the trypticase soy broth containing 1%, 3%, and 5% garlic juice, viable cell number of E. coli O157:H7 was reduced to $2.3{\times}10^2\;CFU/mL$ at 5% from $7{\times}10^8\;CFU/mL$ at the non-treated culture, respectively. The inhibitory effects of the ground beef treated with 3%, 6%, and 10% garlic juice against E. coli O157:H7 was significantly enhanced with approximate 2 log-reduction compared to that of ground beef without garlic. There was no significant difference in the inhibition of E. coli O157:H7 among the groups with different amounts of garlic juice (p<0.05). These results suggest that garlic juice may function well as a natural preservative in food system.