• 한국미생물·생명공학회지
• /
• 제17권1호
• /
• pp.51-55
• /
• 1989

TOL 플라스미드와 NAH 플라스미드는 n-알칸을 자화하는 P. putida KCTC 2405에 접합에 의해 각각의 이동은 가능하나 두 플라스미드는 불화합성에 기인하여 본 균주내에 공존할 수 없었다. TOL plasmid에서 불화합성 체계는 남겨두고 tol 유전자만 이 CAM plasmid내로 transposition 되어 형성된 CAM::TOL* 플라스미드는 NAH 플라스미드와 P. putida KCTC 2405에서 공존할 수 있어 m-toluate, naphthalene, camphor 및 n-alkane(C8-C24)를 분해할 수 있는 P. putida 3SK 균주를 육종하였다. CAM::TOL* 플라스미드는 선택성 배지에서 안정하였으나 비선택성 배지에서는 불안정하였다.

• 미생물학회지
• /
• 제29권6호
• /
• pp.408-415
• /
• 1991

TOL plsmid size of Flavobacterium odoratum SUB53 was estimated as 83 Md and the optimum concentration of m-toluate degradation by TOL plasmid was 5 mM. $H_{2}$ production by Rhodopseudomonas sphaeroides KCTC1425 was largely dependent on nitrogenase activity and showed the highest at 30 mM malate with 7 mM glutamate as nitrogen source. Nitrogenase activities were inhibited by 0.3 mM $NH_{4}^{+}$ions, to be appeared the decrease of $H_{2}$ production. Conjugation of TOL plasmids from F. odoratum SUB53 and Pseudomonas putida mt-2 to R. sphaeroides showed the optimum at the exponential stage of recipient cells in presence of helper plasmid pRK2013. According to the investigation of catechol-1,2-oxygenase (C-1, 2-O) and catechol-2,3-oxygenase (C-2,3-O) activities of R. sphaeroides C1 (TOL SUB53) and C2 (TOL mt-2), the gene for C-2,3-O is located on TOL plasmid and gene for C-1, 2-O on the chromosome of R. sphaeroides. m-Toluate was biodegraded by TOL plasmid in R. sphaeroides C1 and C2, presumably to be produced $H_{2}$ gas from the secondary metabolites of m-toluate.e.

• BMB Reports
• /
• 제56권6호
• /
• pp.326-334
• /
• 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 and Biotechnology
• /
• 제18권5호
• /
• pp.845-851
• /
• 2008

TolC is an outer membrane porin protein and an essential component of drug efflux and type-I secretion systems in Gram-negative bacteria. TolC comprises a periplasmic $\alpha$-helical barrel domain and a membrane-embedded $\beta$-barrel domain. TdeA, a functional and structural homolog of TolC, is required for toxin and drug export in the pathogenic oral bacterium Actinobacillus actinomycetemcomitans. Here, we report the expression of the periplasmic domain of TdeA as a soluble protein by substitution of the membrane-embedded domain with short linkers, which enabled us to purify the protein in the absence of detergent. We confirmed the structural integrity of the TdeA periplasmic domain by size-exclusion chromatography, circular dichroism spectroscopy, and electron microscopy, which together showed that the periplasmic domain of the TolC protein family fold correctly on its own. We further demonstrated that the periplasmic domain of TdeA interacts with peptidoglycans of the bacterial cell wall, which supports the idea that completely folded TolC family proteins traverse the peptidoglycan layer to interact with inner membrane transporters.

• 한국미생물학회:학술대회논문집
• /
• 한국미생물학회 2002년도 추계학술대회
• /
• pp.59-62
• /
• 2002

The role of a TolC homologue in the virulence of Vibrio vulnificus, a marine bacterium causing serious wound infection and fulminant septicemia in persons with underlying conditions, has been studied. TolC, an outer membrane protein, has been implicated in a variety of bacterial functions including export of diverse molecules ranging from large proteins to antibiotics. A homologue of the tolC gene of V. cholerae, which has been shown to be required for bile resistance, cytotoxicity and colonization of this organism, was identified in the partially determined genome sequence of V. vulnificus. To determine the role of TolC in the virulence of V. vulnificus, a TolC-deficient (TD) mutant was isolated by in vivo allelic exchange. Compared with the parent strain, the TD mutant was more sensitive to bile, and much less virulent in mice challenged subcutaneously. This mutant was noncytotoxic to the HEp-2 cells, but its metalloprotease and cytolysin activities in the culture supernatant were comparable to the parent strain. In addition, the resistance of the TD mutant to human serum bactericidal activity as well as its growth in either human or murine blood was not affected. Collectively, our data suggest that TolC may be involved in colonization and/or spread of V. vulnificus to the blood stream, probably by secreting a cytotoxin other than the cytolysin.

• Molecules and Cells
• /
• 제38권2호
• /
• pp.180-186
• /
• 2015

Escherichia coli AcrAB-TolC is a multidrug efflux pump that expels a wide range of toxic substrates. The dynamic nature of the binding or low affinity between the components has impeded elucidation of how the three components assemble in the functional state. Here, we created fusion proteins composed of AcrB, a transmembrane linker, and two copies of AcrA. The fusion protein exhibited acridine pumping activity, suggesting that the protein reflects the functional structure in vivo. To discern the assembling mode with TolC, the AcrBA fusion protein was incubated with TolC or a chimeric protein containing the TolC aperture tip region. Three-dimensional structures of the complex proteins were determined through transmission electron microscopy. The overall structure exemplifies the adaptor bridging model, wherein the funnel-like AcrA hexamer forms an intermeshing cogwheel interaction with the ${\alpha}$-barrel tip region of TolC, and a direct interaction between AcrB and TolC is not allowed. These observations provide a structural blueprint for understanding multidrug resistance in pathogenic Gram-negative bacteria.

• 대한의생명과학회지
• /
• 제10권2호
• /
• pp.153-161
• /
• 2004

Antibiotic resistance is often associated with the production of inner membrane proteins (for example, AcrAB/TolC efflux pump) that are capable to extrude antibiotics, detergents, dyes and organic solvents. In order to evaluate the unknown MarB function of Escherichia coli, especially focused on the function of OmpF porin, several mutants were construted by T4GT7 transduction. MarA plays a major roles in mar (multiple antibiotic resistance) phenotype with AcrAB/TolC efflux pump in E. coli K-12. Futhermore, MarA decreases OmpF porin expression via micF antisense RNA. Expression of acrAB is increased in strains containing mutation in marR, and in those carrying multicopy plasmid expressing marA. MarB protein of E. coli K-12 showed its activity at OmpF porin & TolC protein as target molecule. Some paper reported MarB positively regulates OmpF function. MarA shows mar phenotype, and MarB along with MarA show decreased MIC through OmpF function. By this experiment, MarB could decrease MIC through the OmpF porin & TolC protein as target.

• 한국수산과학회지
• /
• 제51권4호
• /
• pp.444-449
• /
• 2018

Two specimens of Scomberoides tol (99.0 mm and 124.5 mm in total length), belonging to the family Carangidae, order Perciformes were collected from Korean waters using a gape net with wings and a hand net between 2014 and 2017. These specimens are characterized by having the origin of the soft-rayed portion of the dorsal fin just above the origin of the soft-rayed portion of the anal fin, dorsal spines not connected by fin membranes and posterior end of the maxilla and upper jaw not extending beyond the posterior margin of the eye. A comparison of mitochondrial DNA cytochrome c oxidase subunit I sequences indicated that these specimens matched Scomberoides tol (K2P distance, d = 0.002), but differed from other Scomberoides species (6.9-9.1%). This is the first reliable report of Scomberoides tol from Korea.

• 대한의생명과학회지
• /
• 제5권1호
• /
• pp.27-40
• /
• 1999

본 연구는 MarA와 함께 MarB가 AcrAB efflux pump를 목표로 하는지 여부, 그리고 항생제의 균체외 배출기능에 대하여 어떤 조절 작용이 있는지를 항생제 내성검사를 통하여 살펴보았다. 본 연구 결과는 MarB가 MarA와 함께 AcrAB/TolC efflux pump의 항생제 배출기 능을 억제적으로 조절한다는 것을 간접적으로 보여 주었으며, 한편 이미 알려진 대로 MarA는 acrRAB operon의 발현을 전사 수준에 서 positive regulation하므로, MarB는 AcrAB efflux pump의 기능을 억제적으로 조절한다는 것을 암시하고 있다. 그리고 MarA와 함께 MarB단백질의 작용 목표는 AcrAB efflux pump임을 간접적으로 보여주고 있다. 또한 MarB는 MarA와 함께 대장균의 다른 efflux system(들)의 항생제 배출기능에 대해서도 억제적으로 조절할 가능성이 높은 것으로 나타났다.

• Bulletin of the Korean Chemical Society
• /
• 제11권6호
• /
• pp.531-534
• /
• 1990

The reaction of $Os_3(CO)_{10}(NCMe)_2$ with a tungsten alkylidyne $Cp(CO)_2W{\equiv}CTol(Cp={\eta}^5-C_5H_5,\;Tol=p-C_6H_4Me)$ produces the compound, $CpWOs_3(CO)_{11}({\mu}_3-CTol)$(1). The structure of compound 1 can be viewed as one in which the metalla-alkyne, $Cp(CO)_2W{\equiv}CTol$, is ${\mu}_3-{\eta}^2-{\bot}$ (perpendicular) bound to an $Os_3(CO)_9$ fragment. Variable-temperature $^{13}C$ NMR spectra of 1 show no evidence for the analogous rotation of the metalla-alkyne molecule as shown alkyne rotation in $Os_3(CO)_9(C_2Tol_2)$ compound. This lack of the metalla-alkyne fluxionality supports the apparent saturated nature of compound 1.