• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.433-436
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• 1990

The TOL plasmid, pWWO, conjugally transferred from Pseudomonas putida mt-2 was dissociated into TOL* and TOL $\Delta$A in P. putidu PpGl carrying CAM plasmid. The TOL* was integrated into the CAM plasmid, and the resulting plasmid was designated as CAM::TOL*. The introduction of NAH plasmid, belonging to Inc P9 incompatibility group, into P. putida CSTBA carrying CAM::TOLt plasmid and TOL A plasmid did not affect m-toluate catabolism, but resulted in expelling the TOL $\Delta$ plasmid.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.51-55
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• 1989

The conjugally transferred TOL plasmid or NAH plasmid was stably maintained and expressed in n-alkane assimilating Pseudomonas putida KCTC 2405. However, these plasmids were not able to coexist in this strain because of incompatibility. The incompatibility of TOL and NAH plasmid was bypassed using CAM::TOL* plasmid, which was constructed by the transposition of only tol gene without incompatibility system in TOL plasmid into CAM plasmid. p. putida 3SK capable of growing on m-toluate, naphthalene, camphor, and n-alkane(C8-C24) was constructed by the conjugal transfer of NAH plasmid into n-alkane assimilating p. putida SK carrying CAM:: TOL* plasmid. CAM::TOL＊ plasmid in p. putida 3SK was stable on the selective media but unstable on the nonselective media.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.334-341
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• 1989

Toluate degradative plasmid from Pseudomonas cepacia SUB37 was determined to be molecular weight as $79.3\times 10^6$

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.408-415
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• 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.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.321-326
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• 1989

Eighty two bacterial strains have been isolated from five different soil and sewage samples by selective enrichment culture on m-toluate minimal medium. Two of these were identified as Pseudomonas capacia, one as P. putida, one as Yersinia intermedia, and one as Flavobaeterium odoratum. P. cepacia SUB37 appeared to carry plasmid superficially similar to TOL plasmid previously described in p. putida mt-2 and other two plasmids from Flavobacterium odorutum and Y. intermedia larger than that of p. putida mt-2. p. cepacia SUB37 was sensitive to streptomycin but resistant to rifampicin. P. cepacia SUB37 carrying plasmid metabolizes the hydrocarbons to benzoate and toluates via the corresponding alcohols and aldehydes. By the curing experiment, it appears that P. cepacia SUB37 carries TOL plasmid encoding for the enzymes responsible for the catabolism of toluene and xylene via benzoate and the toluates and then by meta pathway in the process of degradation of aromatic hydrocarbons. p. cepacia SUB37 degraded m-toluate rapidly to be very low level when it was fully grown.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.236-242
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• 1990

To develop the new strains of microorganisms having the degradative ability for various aromatic hydrocarbons, the hybrid plasmid pKG2 having the 2,4-Dichlorophenoxyacetic acid(2,4-D) degradative genes, the hybrid plasmid pKG3 containg the naphthalene degradative genes and TOL plasmid were introduced into Pseudomonas putida KUD 12 and P. putida KUP 10 by transformation or conjugation which originally have the degradative ability of the synthetic surfactants and phthalate esters, respectively. From P. putida KUD12, the new strains of P. putida KUD101(pKG2), KUD102(pKG3), KUD103(TOL), and KUD202(pKG3, TOL) were obtained, and KUD106(pKG2), KUD107(pKG3), KUD108(TOL) were originated from the P.putida KUP10. The degradative abilities in P. putida KUD101, KUD102 and KUD107 were similar with those of the original strains. The P. putida KUD103, KUD106 and KUD202 had a little lower and P. putida KUD108 had a better degradative abilitie than those of the original ones. In the case of mixed cultures, the mixed culture of KUD107 and KUD108 had a better degradative abilities than those of the other mixed cultures.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.270-273
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• 1994

Pseudomonas putida 3SK, which was constructed by the conjugal transfet of the $CAM::TOL^{*}$ plasmid of Pseudomonas putida CSnA and the NAH plasmid of Pseudomonas putida KCTC 2403 into n-alkane assimilating Pseudomonas putida KCTC 2405, showed a broad degradation spectrum and floc-forming ability. This strain degraded m-toluic acid, naphthalene, camphor and decane simultaneously. $Hg^{2+}$ at the concentration of 1 ppm in the minimal medium could not inhibit the growth of this strain. The degradation of m-toluic acid by Pseudomonas putida 3SK was not repressed by the easily utilizable compounds, such as glucose and succinate. But, the addition of formalin inhibited the growth of Pseudomonas putida 3SK. After the cultivation of this strain on the artificial wastewater containing m-toluic acid, naphthalene, camphor and decane for 24 hr, the initial COD value (1500) of the artificial wastewater was declined to 300.

• Korean Journal of Microbiology
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• v.29 no.3
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• pp.155-159
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• 1991

Four strains of Pseudomonas putida (NAH), Pseudomonas sp.(TOL), Achromobacter xylosoxidans, and Alcaligenes sp. were compared with their degradative capability of aromatic compounds. All of the bacterial strains were utilized catechol as a sole carbon source for growth, but signigicantly different in degradative properties for 5 other aromatic compounds. Catechol 2, 3-dioxygenase gene from P. putida (NAH) has been cloned and expressed in E. coli. The DNA clone designated pCNU101 contains NAH-derived 6 Kb insert and its physical map was characterized. A subclone (pCNU106) for the catechol dioxygenase gene in pCNU101 contained 2.0kb-DNA insery fragmented by HpaI and ClaI.

• Biomedical Science Letters
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• v.10 no.2
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• pp.153-161
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• 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.

• Journal of Soil and Groundwater Environment
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• v.11 no.5
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• pp.43-50
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• 2006

Bioreporter bacteria, such as recombinant bioluminescent bacteria, have been used for the detection of specific compounds in complex environmental media. In this study, optimum conditions for the preparation and application of deep-freezed and Iyophilized recombinant bioluminescent strain KG1206 were investigated for the future application on contaminated environmental sites. Genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, contains TOL plasmid and the plasmid inserted $P_{m}$, promoter on the upper part of lux gone in vector pUCD615, and m-toluate and benzoate are considered direct inducers for bioluminescence. Optimum conditions determined for the preparation and application of the deep-freezed and lyophilized strain were followings: cryoprotective agent (24% sucrose), lyophilization time (12 hrs), strain concentration ($OD_{600}=0.6$), reconstitution for freezed strain (quick reconstitution at $35^{\circ}C$), reconstitution for lyophilized strain ($3{\sim}6$ hrs exposure on LB medium), carrying conditions (keep at $20^{\circ}C$ after reconstitution). These results demonstrate the feasibility of deep-freezed or lyophilized state of genetically engineered bioluminescent strain for environmental usage.