• Molecules and Cells
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• 제26권1호
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• pp.61-66
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• 2008

Cyclic AMP receptor protein (CRP) is allosterically activated by cAMP and functions as a global transcription regulator in enteric bacteria. Structural information on CRP in the absence of cAMP (apo-CRP) is essential to fully understand its allosteric behavior. In this study we demonstrated interdomain interactions in apo-CRP, using a comparative thermodynamic approach to the intact protein and its isolated domains, which were prepared either by limited proteolysis or using recombinant DNA. Thermal denaturation of the intact apo-CRP, monitored by differential scanning calorimetry, revealed an apparently single cooperative transition with a slight asymmetry. Combined with circular dichroism and fluorescence analysis, the thermal denaturation of apo-CRP could be interpreted as a coupled process involving two individual transitions, each attributable to a structural domain. When isolated individually, both of the domains exhibited significantly altered thermal behavior, thus pointing to the existence of non-covalent interdomain interactions in the intact apo-CRP. These observations suggest that the allosteric conformational change of CRP upon binding to cAMP is achieved by perturbing or modifying pre-existing interdomain interactions. They also underline the effectiveness of a comparative approach using calorimetric and structural probes for studying the thermodynamics of a protein.

• 약학회지
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• 제53권4호
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• pp.194-200
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• 2009

Community-acquired antimicrobial resistant bacteria are an emerging problem whole world-wide. Generally, Hands are main mediator of pathogen transmission as compared with other body parts. So, the purpose of this research was to investigate the microbiological quality and antimicrobial susceptibilities of samples which were collected from the door knob and surface in public- and home-facilities, and also hand. Of total 191 samples, 71 samples (37%) were shown to be of high-level total bacterial count (>$10^8\;CFU/cm^2$). And presence of Staphylococcus and Enteric bacilli was observed in 61 samples (32%) and 76 samples (40%), respectively. Antimicrobial susceptibilities of staphylococcal isolates from the samples were tested for six different antimicrobial agents, which are in wide spread clinical use in Korea, as well as four new antimicrobials, daptomycin, linezolid, quinupristin/dalfopristin and tigecycline. Among staphylococcal isolates, antimicrobial resistance were observed in oxacillin (n=6), mupirocin (n=7), vancomycin (n=1), quinupristin/dalfopristin (n=2) and gentamicin (n=5). Fortunately, all the isolates were susceptible to new antimicrobial such as daptomycin, linezolid and tigecycline. Furthermore 4 Enterococcus faecalis were isolated from four hand samples, and all these isolates exhibited multidrug resistance to four different antimicrobial (oxacillin, quinupristin/dalfopristin, mupirocin, gentamicin). Also, 5 Escherichia coli were isolated from surface in home (n=3), door knob in public facilities (n=1) and hand (n=1). Escherichia coli isolated from hand was high-level resistant to tigecycline ($128{\mu}g$/ml) and gentamicin ($64{\mu}g$/ml).

• 약학회지
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• 제52권6호
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• pp.434-440
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• 2008

The purpose of this research was to investigate the microbiological quality and antimicrobial susceptibilities of the ready-to-eat side dishes, which are were collected from the grocery stores, big markets, and department stores in Seoul throughout November, 2007. Of total 124 samples, presence of staphylococci and enteric bacilli was observed in 38 samples (31%) and 53 samples (43%), respectively. And 30 samples (24%) were shown to be of unsatisfactory quality for total bacterial count (>$10^5$). Antimicrobial susceptibilities of the staphylococci isolated from the side dishes were tested for six different antimicrobial agents, which are in widespread clinical use in Korea, as well as four new antimicrobials, daptomycin, linezolid, quinupristin/dalfopristin and tigecycline. As a result, the staphylococcal isolates were found to be resistant to oxacillin (${MIC}_{90}$, >128 ${\mu}g$/ml), teicoplanin (${MIC}_{90}$, >128 ${\mu}g$/ml), mupirocin (${MIC}_{90}$, >128 ${\mu}g$/ml), linezolid (${MIC}_{90}$, 128 ${\mu}g$/ ml) and quinupristin/dalfopristin (${MIC}_{90}$, 32 ${\mu}g$/ml). Especially, some of the staphylococcal isolates exhibited high level and multi-drug resistance. Moreover, these bacteria were also resistant to new antimicrobials, except tigecycline.

• 생명과학회지
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• 제20권5호
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• pp.676-682
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• 2010

본 연구는 임상검체만이 아닌 주변 생활환경에도 광범위 베타 락탐분해효소를 생성하는 균주(extended-spectrum $\beta$-lactamase, ESBL)가 존재하는지를 확인하고 만약 존재할 경우 그 균주를 분리하고 ESBL유형을 알아보기 위하여 실시되었다. 부산 광안리 하수처리 방류수에서 이중 디스크 확산 검사 결과 양성반응을 나타낸 29균주를 선별하였다. 이중 sodium azide에 내성을 가진 피전달 균주인 Escherichia coli J5에 교차접합이 이루어진 15균주를 대상으로 indole, methyl-red, Voges-Proskauer, Simmon's citrate 시험과 decarboxylase-dihydrolase 및 여러 종류의 당 발효 시험 등 생화학 검사를 실시한 결과 Klebsiella pneumoniae(13균주)와 Escherichia coli(2균주)가 동정되었다. 등전점, PCR, 유전자서열 분석을 실시하여 ESBL 유형을 결정하였다. Klebsiella pneumoniae의 ESBL 생성유형은 SHV-12(4균주)와 SHV-12/TEM-1(9균주)의 2종류로 구분되었고, Escherichia coli의 ESBL 생성유형은 TEM-1(2균주)로 판명되었다.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.761-768
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• 2007

Serratia proteamaculans HY-3 isolated from the digestive tract of a spider produces an extracellular protease named arazyme, with an estimated molecular mass of 51.5 kDa. The purified enzyme was characterized as having high activities at wide pH and temperature ranges. We further characterized biochemical features of the enzymatic reactions under various reaction conditions. The protease efficiently hydrolyzed a broad range of protein substrates including albumin, keratin, and collagen. The dependence of enzymatic activities on the presence of metal ions such as calcium and zinc indicated that the enzyme is a metalloprotease, together with the previous observation that the proteolytic activity of the enzyme was not inhibited by aspartate, cysteine, or serine protease inhibitors, but strongly inhibited by 1,10-phenanthroline and EDTA. The araA gene encoding the exoprotease was isolated as a 5.6 kb BamHI fragment after PCR amplification using degenerate primers and subsequent Southern hybridization. The nucleotide sequence revealed that the deduced amino acid sequences shared extensive similarity with those of the serralysin family of metalloproteases from other enteric bacteria. A gene(inh) encoding a putative protease inhibitor was also identified immediately adjacent to the araA structural gene.

• Journal of Microbiology and Biotechnology
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• 제32권11호
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• pp.1396-1405
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• 2022

Cholera remains a major global public health problem, for which oral cholera vaccines (OCVs) being a valuable strategy. Patients, who have recovered from cholera, develop antibody responses against LPS, cholera toxin (CT), toxin-coregulated pilus (TCP) major subunit A (TcpA) and other antigens; thus, these responses are potentially important contributors to immunity against Vibrio cholerae infection. However, assessments of the efficacy of current OCVs, especially inactivated OCVs, have focused primarily on O-antigen-specific antibody responses, suggesting that more sophisticated strategies are required for inactivated OCVs to induce immune responses against TCP, CT, and other antigens. Previously, we have shown that the toxT-139F allele enables V. cholerae strains to produce CT and TCP under simple laboratory culture conditions. Thus, we hypothesized that V. cholerae strains that express TCP via the toxT-139F allele induce TCP-specific antibody responses. As anticipated, V. cholerae strains that expressed TCP through the toxT-139F allele elicited antibody responses against TCP when the inactivated bacteria were delivered via a mouse model. We have further developed TCP-expressing V. cholerae strains that have been used in inactivated OCVs and shown that they effect an antibody response against TcpA in vivo, suggesting that V. cholerae strains with the toxT-139F allele are excellent candidates for cholera vaccines.

• 지질공학
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• 제34권2호
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• pp.207-216
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• 2024

이 연구는 울릉도 나리분지 추산용천수의 유동 경로를 파악하는 것이다. 칼데라 내부 가장자리를 따라 발달하는 단층과 단열구조들의 발달 방향이 추산용천수 유동 경로에 영향을 주는 요소로 파악되었다. 추산용천수는 함양된 지하수가 동서 방향과 남북 방향의 단열대를 따라 추산용출소 방향으로 흐른다. 이와 더불어 북북동 방향으로 경사진 칼데라의 부등함몰의 영향으로 북동 내지 북북동 방향으로 흐르는 것으로 파악되었다. 장내세균과인 Entrobacteriaceae을 이용하여 나리지하수 쪽의 물이 추산용출소 방향으로 유동하는 것으로 파악되었다. 다만, 분석에 사용된 시료수가 매우 적어 성인계곡수로부터 나리분지와 추산용출소까지의 유동경로를 전체적으로 파악할 수 없다는 제한성이 있다.

• 한국가금학회지
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• 제38권3호
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• pp.173-179
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• 2011

본 연구는 복합 생균제를 사료 공급을 통하여 갈색 산란계(Hy-Line Brown)에 산란 초기 21~40주, 산란 중기인 41~65주로 나누어 산란계의 생산성과 산란에 미치는 영향을 조사하였다. 복합 생균제를 생산하고 생균제를 구성하는 균주의 특성에 대하여 내산성(pH 2.5), 내담즙성(Oxgall 0.3%) 및 특정 병원균에 대한 항균력을 시험하였고, 산란 시기별로 분변 내 미생물의 변화를 조사하였다. 생균제는 4종의 복합생균으로 각각의 대두박 기질에 의한 고상식 발효물로서 원료에서 생산까지 9단계 과정을 거쳐 생산하였다. 복합 생균제를 구성하는 Bacillus subtilis, Lactobacillus plantarum, Enterococcus faecium에서 높은 내산성, 내담즙성, 항균력 등이 나타나는 것을 확인하였으며, Saccharomyces cerevisiae는 항균력이 낮은 것으로 나타났다. 산란 생산성은 급여 후 대조구에 비해 높은 산란율을 보였으나, 0.1% 급여구와 0.2% 급여구 사이의 유의차는 없었다(P>0.05). 난중에 있어서 산란 초기 21~40주는 대조구와 유의차를 보였으나, 0.1% 급여구와 0.2% 급여구간에는 유의차가 인정되지 않았다(P>0.05). 그러나 산란 중기에서 0.2% 급여구와 0.1% 급여구 사이에 유의차를 보였으며, 대조구와도 유의성 각각 인정되었다(P<0.05). 폐사율은 대조구와 비교하였을 때 0.2% 급여구에서 폐사율이 격감되는 유의차가 있었고(P<0.05) 산란 시기별로는 산란 초기에 0.2% 급여구가 폐사율 감소를 보였으나, 산란 중기에는 0.1% 급여구와의 유의차가 나타나지 않았다. 사료 섭취량은 산란 전기에는 유의성이 없었으나, 산란 후기의 0.2% 급여구에서 유의적인 증가가 인정되었다(P<0.05). 분변 미생물 조사에서 복합 생균제 급여 후 0.1% 급여구의 전기, 중기 모두 Lactobacillus가 유의적으로 증가되었으나, Coli form은 중기에 0.1%, 0.2% 급여구에서 각각 유의적으로 감소하는 경향을 나타내었다(P<0.05). 결론적으로 산란계 사료 내 복합 생균제의 첨가는 산란율과 난중의 향상과 분변 미생물의 정상적인 변화로 폐사율을 감소시키는 것으로 사료된다.