• KSBB Journal
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• 제27권4호
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• pp.257-262
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• 2012

The gene encoding Thermus thermophilus HJ6 laccase (Tt-laccase) was cloned, sequenced, and comprised of 1,389 nucleotides encoding a protein (462 amino acids) with a predicted molecular mass of 51,049 Da. The deduced amino acid sequence of Tt-laccase showed 99.7% and 44.3% identities to the Thermus thermophilus HB27 laccase and Synechococcus sp. RS9917 laccase, respectively. Tt-laccase gene was expressed as a fusion protein with six histidine residues in E. coli Rosetta-gami (DE3) cells, and the recombinant protein was purified to homogeneity. UV-Vis spectrum analysis revealed that the enzyme has copper atoms, a type I Cu(II) and a type III binuclear Cu(II). The optimum pH for the oxidation of guaiacol was 5.0 and the optimum temperature was $90^{\circ}C$ The half-life of heat inactivation was about 120 min at $90^{\circ}C$ The enzyme reaction was inhibited by sodium azide, L-cystein, EDTA, dithiothreitol, tropolone, and kojic acid. The enzyme oxidized various known laccase substrates, its lowest $K_m$ value being for 4-hydroxyindole, highest $k_{cat}$ value for syringaldazine, and highest $k_{cat}/K_m$ for guaiacol.

• 한국동물위생학회지
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• 제45권2호
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• pp.117-124
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• 2022

In the event of an outbreak of a livestock epidemic, it has been considered that the existing burial-centered carcass disposal method should be improved ecofriendly for prevention of leachate and odors from burial basically in regard of pathogen inactivation. Therefore, the aim of this study is whether it was possible to treat the carcass of cattle and chickens using the chemical carcass treatment method. It was conducted to establish detailed treatment standards for the chemical treatment method of cattle and chicken carcasses based on the results of the proof of the absence of infectious diseases in cattle chickens. After inoculating cattle carcass with 10 pathogens (foot and mouth disease virus, bovine viral diarrhea virus, Mycobacterium bovis, Mycobacterium avium subsp. Paratuberculosis, Brucella abortus, Bacillus anthracis, Clostridium chauvoei, Clostridium perfringens, Escherichia coli, and Salmonella Typhimurium) and chicken carcasses with low pathogenic avian influenza virus, Clostridium perfringens type C, E. coli and Salmonella Typhimurium, these were treated at 90℃ for 5 hours in a potassium hydroxide liquid solution corresponding to 15% of the body weight. This method liquefies all cadaveric components and inactivates all inoculated pathogens by PCR and culture. Based on these results, it was possible to prove that chemical treatment of cattle and chicken carcasses is effective in killing pathogens and is a safe method without the risk of disease transmission. The chemical treatment method of livestock carcasses can be suggested as an alternative to the current domestic burial-centered livestock carcass treatment method, preventing environmental pollution, and contributing to public health.

• 한국식품저장유통학회지
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• 제14권5호
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• pp.481-486
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• 2007

본 연구는 감마선 조사와 실크 펩타이드 병용처리가 세균의 방사선 감수성 및 햄버거 패티의 저장 안전성에 미치는 영향을 평가하기 위해 실시되었다. 멸균된 햄버거 패티에 접종된 4종의 세균에 대한 감마선 $D_{10}$ 값은 E. coli 0.25 kGy, L. ivanovii 0.50 kGy, S. typhimurium 0.55 kGy, C. sporogenes 1.35 kGy 이였다. 실크 펩타이드 5% 첨가 햄버거 패티에 접종된 S. typhimurium의 $D_{10}$ 값은 대조구의 0.558 kGy에서 0.53 kGy로 6% 낮아지는 것으로 나타났다. 한편, $30^{\circ}C$ 가속 저장조건에서 햄버거 패티의 미생물 생육도를 측정한 결과 7 kGy이상의 감마선 조사에 의해 저장기간 동안 미생물이 검출되지 않았으며, 실크 펩타이드 7% 첨가시 대조구에 비해 약 1 log cycle의 미생물 생육 감소를 확인하였다. 따라서 실크 펩타이드 첨가는 방사선 조사와 같이 다른 식품 살균기술과 함께 사용한다면 식품의 초기오염 미생물을 제어하여 저장 안정성을 확보하는데 도움이 되는 것으로 판단되었다.

• Restorative Dentistry and Endodontics
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• 제27권5호
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• pp.463-472
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• 2002

Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-$\alpha$ from immune cells. Although rnonocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-$\alpha$. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)$_2$ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-$\alpha$, and it was compared with Escherichia coli LPS. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 $\mu\textrm{g}$/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)$_2$ at 37$^{\circ}C$ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4$\times$10$^6$ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10$\mu\textrm{g}$/ml) for 24 hours at 37$^{\circ}C$ in 5% $CO_2$ incubator. The supernatants of cells were collected and the levels of IL-1$\alpha$, IL=1$\beta$ and TNF-$\alpha$ were measured by enzyme-linked immunosorbent assay. The results were as follows ; 1. The levels of IL-1$\alpha$, IL-1$\beta$, TNF-$\alpha$ from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05) 3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with p. endodontalis LPS were significantly lower than those released from PMN stimulated with E coli LPS (p<0.05).