• Microbiology and Biotechnology Letters
• /
• v.16 no.5
• /
• pp.427-431
• /
• 1988

The simultaneons mineralization of aniline and other secondary carbon sources by Pseudomonas testosteroni 6Fl were evaluated by the lag time and the enzyme induction level. The lag time for aniline mineralization by P. testosteroni 6Fl was 7 hours, whereas the lag time for aniline and readily utilizable secondary substrates were 1-3 hours. This stimulated degradation resulted from the simultaneous use of secondary substrates and aniline, the increased rate of enzyme induction, and the in-creased rate of the cell growth. The enzyme induction level of P. testosteron 6F1 were varied according to the kinds of secondary substrate.

• Korean Journal of Microbiology
• /
• v.21 no.4
• /
• pp.207-212
• /
• 1983

Isolated Gram-negative bacteria, being capable of degrading toxic, recalcitrant, and pigment-producing furfural, were tentatively identified as Pseudomonas testosteroni, Pseudomonas maltophilia, Klebsiella Pneumoniae, and Pseudomonas fluorescens. They exhibited synergistic effects between P. testosteroni and the others in the degradation of colourproducing furfural. Synergistic effects and possible sequence of its degradation were attempted by manometric technique. P. testosteroni could degrade furfural to decolourize it and produce ninhydrin-reaction postive substance (NPS) which could be utilized by P. maltophilia and K. pneumoniae and the latter two bacteria could ,degrade furfural to 2-furoic acid as an oxidized form. Finally 2-furoic acid was further oxidized by P. fluorescens. Once NPS and 2-furoic acid were produced, the degradation efficiency was enhanced by competing four bacteria against furfural and 2-furoic acid.

• Korean Journal of Microbiology
• /
• v.25 no.1
• /
• pp.67-72
• /
• 1987

Cellular fatty acid composition of eight strains, indluding six strains of Comamonas terrigena, and two type strains of Pseudomonas acidovorans, and P. testosteroni was determined by gas-liquid chromatography. Almost the same composition was found in all the strains tested, and hexadecanoic acid, hexadecenoic acid, and octadecenoic acid were accounted more than 70% of total fatty acid. However, P. testosteroni differed from C. terrigena and P. acidovorans by the presence of comparatively large amonuts of 2-hydroxy-hexadecanoic acid, and C. terrigena contained three to eight times as much tetradecanoic acid in P. acidovorans and P. testosteroni. According to the similarity values calculated on the basis of fatty acid composition, C. terrigena strains were divided into three groups differentiated in the requirement of growth factors, and C. terrigena, P. acidovorans, and P. testosteroni strains occupied separate position each other in the dendrogram.

• Applied Biological Chemistry
• /
• v.32 no.3
• /
• pp.315-320
• /
• 1989

Bacteria utilizing naphthalene as a sole carbon source for growth were isolated and identified and code named as Acinetobacter calcoaceticus R-88, Pseudomonas testosteroni R-87 and Pseudomonas putida R-89. Among these isolates, A. calcoaceticus R-88 found most effective in utilizing naphthalene. The optimal pH, temperature and concentration of naphthalene was 7.0, $30^{\circ}C$ and 10mM, respectively. The strain degraded naphthalene to salicylic acid as an intermediate. And the strain showed to be resistant to ampicillin, tetracyclin, chloramphenicol and kanamycin. A. calcoaceticus R-88 harbored plasmid DNA which was believed to be involved in naphthalene degradation.

• Proceedings of the Korean Biophysical Society Conference
• /
• 1999.06a
• /
• pp.36-36
• /
• 1999

Bacterial Δ-3-ketosteroid isomerase (KSI) catalyzes the conversion of Δ-to Δ-3-ketosteroids via enolate formation, which is also found in the synthesis of all steroid hormones in mammals. In Pseudomonas testosteroni, KSI Asp38 (pKa ~ 4.7) was identified as the general base which abstracts the steroid C4b-H (pKa ~ 12.7) to form the dienolate intermediate.(omitted)

• Microbiology and Biotechnology Letters
• /
• v.31 no.2
• /
• pp.177-181
• /
• 2003

Eleven bacterial strains which are able to utilize terephthalic acid as a carbon and an energy source for growth were isolated from the soil of 7 water quality evaluation points in Kyonggi area of Korea. Phthalic acid isomer degrading activity of the isolates from the 4 contaminated points was higher than those from the 3 clean points. Among 11 isolates, 4 isolates which have high terephthalic acid degrading activity and degrade two phthalic acid isomers were identified by partal 16S rDNA sequence determination. One of them was identified as Pseudomonas putida, and the others as Comamonas testosteroni. Thus a large number of phthalic acid isomer degrading bacteria in domestic soil were inferred as C. testosteroni. On the basis of these results, the PCR detection of C. testosteroni in soil was applied to monitor soil contamination by phthalic acid isomers. The DNA of C. test-osteroni extracted from 4 g soil was directly detected by PCR with C. testosteroni specific primer pair. The amount of PCR products was different according to sampling sites and more PCR products were obtained from contaminated sites than those from clean sites (Gulpo-chun＞Anyang-chun＞Hwangguji-chun>Shin-chun＞Huk-chun＞Pukhan-river>Kapyeong-chun). This result was coincided with that of the viable cell counts for terephthalic acid degrading bacteria.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.12
• /
• pp.1995-1999
• /
• 2006

Pseudomonas sp. K82 cultured in p-hydroxybenzoate induces protocatechuate 4,5-dioxygenase (PCD 4,5) for p-hydroxybenzoate degradation. In this study, a 6.0-kbp EcoR1 fragment containing p-hydroxybenzoate degradation genes was cloned from the genome of Pseudomonas sp. K82. Sequence analysis identified four genes, namely, pcaD, pcaA, pcaB, and pcaC genes known to be involved in p-hydroxybenzoate degradation. Two putative 4-hydroxyphenylpyruvate dioxygenases and one putative oxidoreductase were closely located by the p-hydroxybenzoate degradation genes. The gene arrangement and sequences of these p-hydroxybenzoate degradation genes were similar to those of Comamonas testosteroni and Pseudomonas ochraceae. PcaAB (PCD4,5) was overexpressed in the expression vector pGEX-4T-3, purified using a GST column, and confirmed to have protocatechuate 4,5-dioxygenase activity. The N-terminal amino acid sequences of overexpressed PCD4,5 were identical with those of purified PCD4,5 from Pseudomonas sp. K82.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.5
• /
• pp.700-705
• /
• 2003

Pseudomonas sp. S-47 is capable of degrading benzoate and 4-chlorobenzoate as well as catechol and 4-chlorocatechol via the meta-cleavage pathway. The three enzymes of 2-oxopenta-4-enoate hydratase (OEH), acetaldehyde dehydrogenase (acylating) (ADA), and 2-oxo-4-hydroxypentonate aldolase (HOA) encoded by xylJQK genes are responsible for the three steps after the meta-cleavage of catechol. The nucleotide sequence of the xylJQK genes located in the chromosomal DNA was cloned and analyzed. GC content of xylJ, xylQ, and xylK was 65% and consisted of 786, 924, and 1,041 nucleotides, respectively. The deduced amino acid sequences of xylJ, xylQ, and xylK genes from Pseudomonas sp. S-47 showed 93%, 99%, and 99% identity, compared with those of nahT, nahH, and nahI in Pseudomonas stutzeri An10. However, there were only about 53% to 85% identity with xylJQK of Pseudomonas putida mt-2, dmpEFG of P. putida CF600, aphEFG of Comamonas testosteroni TA441, and ipbEGF of P. putida RE204. On the other hand, the xylLTEGF genes located upstream of xylJQK in the strain S-47 showed high homology with those of TOL plasmid from Pseudomonas putida mt-2. These findings suggested that the xylLTEGFIJQK of Pseudomonas sp. S-47 responsible for complete degradation of benzoate and then catechol via the meta-pathway were phylogenetically recombinated from the genes of Pseudomonas putida mt-2 and Pseudomonas stutzeri An10.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.2
• /
• pp.302-311
• /
• 2004

Comamonas testosteroni (formerly Pseudomonas sp.) NCIMB 10643 can grow on biphenyl and alkylbenzenes $(C_2-C_7)$ via 3-substituted catechols. Thus, to identify the genes encoding the degradation, transposon-mutagenesis was carried out using pAG408, a promoter-probe mini-transposon with a green fluorescent protein (GFP), as a reporter. A mutant, NT-1, which was unable to grow on alkylbenzenes and biphenyl, accumulated catechols and exhibited an enhanced expression of GFP upon exposure to these substrates, indicating that the gfp had been inserted in a gene encoding a broad substrate range catechol 2,3-dioxygenase. The genes (2,826 bp) flanking the gfp cloned from an SphI-digested fragment contained three complete open reading frames that were designated bphCDorfl. The deduced amino acid sequences of bphCDorfl were identical to 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC), 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase (BphD), and OrfI, respectively, that are all involved in the degradation of biphenyl/4-chlorobiphenyl (bph) by Ralstonia oxalatica A5. The deduced amino acid sequence of the orfl revealed a similarity to those of outer membrane proteins belonging to the OmpW family. The introduction of the bphCDorfl genes enabled the NT-l mutant to grow on aromatic hydrocarbons. In addition, PCR analysis indicated that the DNA sequence and gene organization of the bph operon were closely related to those in the bph operon from Tn4371 identified in strain A5. Furthermore, strain A5 was also able to grow on a similar set of alkylbenzenes as strain NCIMB 10643, demonstrating that, among the identified aromatic hydrocarbon degradation pathways, the bph degradation pathway related to Tn4371 was the most versatile in catabolizing a variety of aromatic hydrocarbons of mono- and bicyclic benzenes.

• Journal of Microbiology
• /
• v.42 no.2
• /
• pp.152-155
• /
• 2004

Pseudomonas sp. K82 has been reported to be an aniline-assimilating soil bacterium. However, this strain can use not only aniline as a sole carbon and energy source, but can also utilize benzoate, p-hydroxybenzoate, and aniline analogues. The strain accomplishes this metabolic diversity by using dif-ferent aerobic pathways. Pseudomonas sp. K82, when cultured in p-hydroxybenzoate, showed extradiol cleavage activity of protocatechuate. In accordance with those findings, our study attempted the puri-fication of protocatechuate 4,5-dioxygenase (PCD 4,5). However the purified PCD 4,5 was found to be very unstable during purification. After Q-sepharose chromatography was performed, the crude enzyme activity was augmented by a factor of approximately 4.7. From the Q-sepharose fraction which exhibited PCD 4,5 activity, two subunits of PCD4,5 (${\alpha}$ subunit and ${\beta}$ subunit) were identified using the N-terminal amino acid sequences of 15 amino acid residues. These subunits were found to have more than 90％ sequence homology with PmdA and PmdB of Comamonas testosteroni. The molecular weight of the native enzyme was estimated to be approximately 54 kDa, suggesting that PCD4,5 exists as a het-erodimer (${\alpha}$$_1$${\beta}$$_1$). PCD 4,5 exhibits stringent substrate specificity for protocatechuate and its optimal activity occurs at pH 9 and 15 $^{\circ}C$. PCR amplification of these two subunits of PCD4,5 revealed that the ${\alpha}$ subunit and ${\beta}$ subunit occurred in tandem. Our results suggest that Pseudomonas sp. K82 induced PCD 4,5 for the purpose of p-hydroxybenzoate degradation.