• BMB Reports
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• 제56권6호
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• pp.326-334
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• 2023

Antibiotic resistance (AR) is a silent pandemic that kills millions worldwide. Although the development of new therapeutic agents against antibiotic resistance is in urgent demand, this has presented a great challenge, especially for Gram-negative bacteria that have inherent drug-resistance mediated by impermeable outer membranes and multidrug efflux pumps that actively extrude various drugs from the bacteria. For the last two decades, multidrug efflux pumps, including AcrAB-TolC, the most clinically important efflux pump in Gram-negative bacteria, have drawn great attention as strategic targets for re-sensitizing bacteria to the existing antibiotics. This article aims to provide a concise overview of the AcrAB-TolC operational mechanism, reviewing its architecture and substrate specificity, as well as the recent development of AcrAB-TolC inhibitors.

• Journal of Microbiology
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• 제39권1호
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• pp.62-66
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• 2001

Unlike eukaryotic efflux pumps energized by ATPase bacterial efflux pumps are energized by the proton motive force. That is the reason why CCCP, an inhibitor of proton motive forcer is widely used to study the bacterial efflux pump. In many cases, efflux systems have been observed only in quinolone-resistant bacteria. Most of the quinolone-susceptible strains have been found to maintain little efflux pump. However some susceptible bacteria skewed the increased intracellular quinolone concentration only at a low concentration (0.01 or 0.1 mM) but net at a high concentration (1 mM) of CCCP. If bacterial cells were killed at high concentrations of CCCP and lost the integrity of their membranes, the intracellular quinolone would leak out from cells with no efflux system. The efflux pump system in the quinolone-susceptible strains could net be detected at the same concentration used for resistant bacteria. To test this hypothesist the intracellular quinolone concentration in the quinolone-susceptible and -resistant strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus was assayed at various concentrations of CCCP. Since the effect of CCCP is very rapid, the survival of bacteria was observed by assaying the DNA synthesis in 5 min. In the case of E. coli, but not P. aeruginosa or S. aureus, the quinolone susceptible strain was more susceptible to CCCP than the quinolone resistant ones, especially when the incubation with CCCP was extended. Decrease of the intracellular quinolone concentration resulted in a false result-no or weak efflux system in the quinolone susceptible strains. Results suggested that the concentration of CCCP should be optimized in order to detect the efflux system in the quinolone susceptible strains of E. coli.

• 대한의생명과학회지
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• 제27권2호
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• pp.59-68
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• 2021

Multidrug-resistant strain of Acinetobacter baumannii (MDRAB) is an emerging pathogen in health care facilities, preventing MDRAB is a public health concern. We conducted this experiment on a clinical isolate of A. baumannii with two main goals: the role of the efflux pump system in the stress provision of carbapenem and the response to the transcription level of the efflux pump gene. A total of 34 strains of A. baumannii was isolated from the Yangsan Hospital of Pusan National University. First, when we compared and observed the expression of the efflux pump gene and antibacterial resistance to carbapenem, a strong correlation was observed between carbapenem resistance and overexpression of adeB (P=0.0056). Second, a correlation between the efflux pump and concentration gradient and tolerance to carbapenem stress at the AdeABC efflux pump genes transcription level was confirmed. Our results revealed that the expression of the AdeABC efflux pump is an important resistance determinant in obtaining antibiotic resistance of the carbapenem group in A. baumannii.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.658-662
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• 2008

Effects of pH shock on the secretion system of S. coelicolor A3(2) have been investigated at a transcriptional level by using DNA microarrays. Actinorhodin secretion was observed to be highly enhanced when an acidic-pH shock was applied to surface grown cultures of S. coelicolor A3(2). In this culture, a gene of actVA-orf1 encoding a putative efflux pump or transporter protein for actinorhodin was strongly upregulated. A major number of efflux pumps for other metabolites and a major number of secretion proteins for protein secretion were also observed to be upregulated with pH shock. The secretion of actinorhodin was observed to be remarkably enhanced in liquid culture as well.

• Molecules and Cells
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• 제42권12호
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• pp.850-857
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• 2019

The Gram-negative opportunistic pathogen, Pseudomonas aeruginosa, has multiple multidrug efflux pumps. MexT, a LysR-type transcriptional regulator, functions as a transcriptional activator of the MexEF-OprN efflux system. MexT consists of an N-terminal DNA-binding domain and a C-terminal regulatory domain (RD). Little is known regarding MexT ligands and its mechanism of activation. We elucidated the crystal structure of the MexT RD at 2.0 Å resolution. The structure comprised two protomer chains in a dimeric arrangement. MexT possessed an arginine-rich region and a hydrophobic patch lined by a variable loop, both of which are putative ligand-binding sites. The three-dimensional structure of MexT provided clues to the interacting ligand structure. A DNase I footprinting assay of full-length MexT identified two MexT-binding sequence in the mexEF-oprN promoter. Our findings enhance the understanding of the regulation of MexT-dependent activation of efflux pumps.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.430-435
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• 2013

Multidrug resistance, especially multidrug efflux mechanisms that extrude structurally unrelated cytotoxic compounds from the cell by multidrug transporters, is a serious problem and one of the main reasons for the failure of therapeutic treatment of infections by pathogenic microorganisms as well as of cancer cells. Streptococcus mutans is considered one of the primary causative agents of dental caries and periodontal disease, which comprise the most common oral diseases. A fragment of chromosomal DNA from S. mutans KCTC3065 was cloned using Escherichia coli KAM32 as host cells lacking major multidrug efflux pumps. Although E. coli KAM32 cells were very sensitive to many antimicrobial agents, the transformed cells harboring a recombinant plasmid became resistant to several structurally unrelated antimicrobial agents such as tetracycline, kanamycin, rhodamin 6G, ampicillin, acriflavine, ethidium bromide, and tetraphenylphosphonium chloride. This suggested that the cloned DNA fragment carries a gene encoding a multidrug efflux pump. Among 49 of the multidrug-resistant transformants, we report the functional gene cloning and characterization of the function of one multidrug efflux pump, namely MdeA from S. mutans, which was expressed in E. coli KAM32. Judging from the structural and biochemical properties, we concluded that MdeA is the first cloned and characterized multidrug efflux pump using the proton motive force as the energy for efflux drugs.

• 미생물학회지
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• 제49권2호
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• pp.126-130
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• 2013

원유시료에서 분리한 장알균속 세균 245균주의 항생제 내성에 관여하는 다중약물 유출 펌프 유전자 분포도와 항생제 내성 패턴을 연구하였다. 그 결과 245 장알균속 균주의 ampicillin에 대한 내성률은 44.1%, erythromycin에 대한 내성률은 79.2%, tetracycline에 대한 내성률은 76.3%, chloramphenicol에 대한 내성률은 36.3%였으며 vancomycin과 ciprofloxacin에 대해서는 모두 감수성임을 알 수 있었다. 내성 관련 유전자 중 MFS타입의 eme(A)는 82.1%의 장알균에 분포하였으며, ABC 타입의 유전자인 efr(A)는 72.7%, efr(B)는 77.1%, lsa는 71.8%의 장알균에 분포하였다. 특히 이러한 유전자의 기원 세균인 Enterococcus faecalis의 경우 eme(A)는 92.5%, efr(A)는 87.4%, efr(B)는 88.4%, lsa는 88.4%의 분포도를 나타내었다. 한편 동일한 장알균속이지만 종이 다른 장알균에서 eme(A)는 E. faecium 4균주, E. avium 7균주, E. durans 4균주 및 E. raffinosus 2균주에 분포하고 있었다. efr(A)는 E. faecium 2균주와 E. durans 2균주에 분포하였으며, efr(B)는 E. faecium 4균주, E. avium 5균주 및 E. durans 4균주에 분포하였다. 본 연구는 우리나라 원유시료에서 분리한 장알균속의 여러 종의 세균에서 동일한 다중약물 유출 펌프(multidrug efflux pump)의 분포에 대한 첫 번째 보고라 사료되며 E. faecalis 이외의 장알균속에서 이러한 유전자의 분포는 서로 다른 종간의 유전자의 수평적인 이동의 가능성을 시사한다.

• 한국축산시설환경학회지
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• 제9권1호
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• pp.45-56
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• 2003

$TAO^{TM}$ System is an auto-heated thermophilic aerated digestion process using a proprietary microbe called as a Phototropic Bacteria (PTB). High metabolic activity results in heat generation, which enables to produce a pathogen-free and digested liquid fertilizer at short retention times. TAO$^{TM}$ system has been developed to reduce a manure volume and convert into the liquid fertilizer using swine manure since 1992. About 100 units have been installed and operated in Korean swine farms so far. TAO$^{TM}$ system consists of a reactor vessel and ejector-type aeration pumps and foam removers. The swine slurry manure enters into vessel with PTB and is mixed and aerated. The process is operated at detention times from 2 to 4 days and temperature of 55 to $65^{\circ}C$. Foams are occurred and broken down by foam removers to evaporate water contents. Generally, at least 30% of water content is evaporated, 99% of volatile fatty acids caused an odor are removed and pathogen destruction is excellent with fecal coliform, rotavirus and salmonella below detection limits. The effluent from TAO$^{TM}$ system, called as the "TAO EFFLUX", is screened and has superb properties as a fertilizer. Normally N-P-K contents of screened TAO Efflux are 4.7 g/L, 0.375 g/L and 2.8 g/L respectively. The fertilizer effect of TAO EFFLUX compared to chemical fertilizer has been demonstrated and studied with various crops such as rice, potato, cabbage, pumpkin, green pepper, parsley, cucumber and apple. Generally it has better fertilizer effects and excellent soil fertility improvement effects. Moreover, the TAO EFFLUX is concentrated through membrane technology without fouling problems for a cost saving of long distance transportation and a commercialization (crop nutrient commodity) to a gardening market, for example.

• 한국식품위생안전성학회지
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• 제35권2호
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• pp.103-108
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• 2020

최근 최소한으로 가공된 안전한 식품에 대한 소비자의 수요가 기하급수적으로 증가하고 있다. 이러한 이유로 많은 식품가공 업체에서는 식품안전을 강화하고 유통기한을 연장하기 위한 최소한의 가공공정 중 허들기술(hurdle technology)을 적용하고 있다. 한편, 연구에 따르면 식품에 함유된 병원균을 비활성화하기 위한 공정 및 방법들은 식중독세균들의 스트레스 적응 메커니즘을 촉발시켜 심지어 후속 치료로 부터 교차 보호를 준다. 또한, 항생제와 제초제 사용과 같은 일상적인 농장 관행은 항생제 내성을 가진 병원균의 생성을 초래할 수 있다. 이러한 항생제 내성 박테리아는 식품 처리과정과 관련된 스트레스에 내성을 가질 수 있고 가공 식품에서 생존할 수 있는 가능성을 높일 수 있다. 이 리뷰에서는 식품가공과 관련된 스트레스와 항생제 내성의 상관관계에 대해 논의한다. 또한, 항균성 화합물 및 기타 식품 처리 관련 스트레스에 대한 교차 보호 수단으로서 시그마 인자(sigma factors), SOS 반응 경로(SOS response pathways) 및 유출 펌프(efflux pumps)의 사용과 같은 분자유전학적 기작에 대해서도 논의한다.

• 생명과학회지
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• 제26권12호
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• pp.1392-1399
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• 2016

Jaspine B는 석회해면류에서 추출된 sphingosine유도체로 인간 암세포에서의 항암활성이 보고되었다. 그러므로 본 연구는 다양한 인간 암세포주에서 항암활성을 비교하고, 암세포주에서의 Jaspine B의 농도를 측정하여 항암 활성과의 연관성을 확인하고자 하였다. 항암활성은 MTT 방법을 이용하여 측정하였고, $EC_{50}$ 값으로 표현하였다. 암세포주내 Jaspine B의 농도는 LC-MS/MS를 이용하여 분석하였다. 항암활성은 세포주마다 다양하게 나타났는데, 유방암과 흑색종 세포주에서 항암활성이 높게 나타났으며($EC_{50}$ 각각 $2.3{\mu}M$과 $2.6{\mu}M$), 신장암세포주에서는 $EC_{50}$ 값이 $29.4{\mu}M$이었다. 암세포주에서의 $EC_{50}$ 값은 동일한 세포에서의 Jaspine B 농도와 높은 상관성을 나타내었으며(r=0.838), 암세포내 약물농도를 조절하는 것으로 잘 알려진 P-glycoprotein과 breast cancer resistance protein 등의 배출수송계와는 관련이 없음을 확인하였다. 이상의 결과는 세포내 약물농도를 높게 유지하는 것이 항암활성에 매우 중요하며, 세포내 약물농도가 암세포주에 따라 다른 약효를 보이는 원인으로 사료된다.