• Journal of Microbiology
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• v.38 no.4
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• pp.245-249
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• 2000

Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.483-489
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• 2004

Pseudomonas sp. S-47 is a bacterium capable of degrading benzoate as well as 4-chlorobenzoate (4CBA). Benzoate and 4CBA are known to be degraded via a meta-cleavage pathway characterized by a series of enzymes encoded by xyl genes. The meta-cleavage pathway operon in Pseudomonas sp. S-47 encodes a set of enzymes which transform benzoate and 4CBA into TCA cycle intermediates via the meta-cleavage of (4-chloro )catechol to produce pyruvate and acetyl-CoA. In the current study, the meta-pathway gene cluster was cloned from the chromosomal DNA of S-47 strain to obtain pCS1, which included the degradation activities for 4CBA and catechol. The genetic organization of the operon was then examined by cloning the meta-pathway genes into a pBluescript SKII(+) vector. As such, the meta-pathway operon from Pseudomonas sp. S-47 was found to contain 13 genes in the order of xylXYZLTEGFlQKIH. The two regulatory genes, xylS and xylR, that control the expression of the meta-pathway operon, were located adjacently downstream of the meta-pathway operon. The xyl genes from strain S-47 exhibited a high nucleoside sequence homology to those from Pseudomonas putida mt-2, except for the xylJQK genes, which were more homologous to the corresponding three genes from P. stutzeri AN10. One open reading frame was found between the xylH and xylS genes, which may playa role of a transposase. Accordingly, the current results suggest that the xyl gene cluster in Pseudomonas sp. S-47 responsible for the complete degradation of benzoate was recombined with the corresponding genes from P. putida mt-2 and P. stutzeri AN10.

• Journal of Microbiology
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• v.35 no.4
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• pp.295-299
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• 1997

Pseudomonas sp. S-47 is capable of transforming 4-chlorobenzoate to 4-chlorocatechol which is subsequently oxidized bty meta-cleavage dioxygenase to prodyce 5-chloro-2-hydroxymuconic semialdehyde. Catechol 2,3-dioxygenase (C23O) produced by Pseudomonas sp. S-47 was purified and characterized in this study. The C23O enzyme was maximally produced in the late logarithmic growth phase, and the temperature and pH for maximunm enzyme activity were $30{\sim}35^{\circ}C$ and 7.0, respectively. The enzyme was purified and concentrated 5 fold from the crude cell extracts through Q Sepharose chromatography and Sephadex G-100 gel filtration after acetone precipitation. The enzyme was identified as consisting of 35 kDa subunits when analyzed by SDS-PAGE. The C23O produced by Pseudomonas sp. S-47 was similar to Xy1E of Pseudomonas putida with respect to substrate specificity for several catecholic compounds.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.111-114
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• 2000

Streptomyces are widespread in nature and play a very important role in the biosynthesis as well as biodegradation of natural and unnatural aromatic compounds. Both qualitatively and quantitatively through TLC and UV spectrophotometric assays, it was observed that the thermophilic soil bacteria S. setonii (ATCC 39116), which can utilize a benzoate as a sole carbon and energy source in a minimal liquid culture, was not very sensitive to the benzoate concentation and to the culture conditions such as the pH and temperature. The in vitro conversion of a catechol to a cis, cis-muconic acid by a crude S. setonii lysate implies that the aromatic ring cleavage by S. setonii is initiated by a thermostable catechol-1,2-dioxygenase, the key enzyme in the ortho-cleavage pathway of aromatic compound biodegradation. Unlike non-degrading S. lividans, S.setonii was also highly resistant to other similar hazardous aromatic compounds, exhibiting almost no adverse effect on its growth in a complex liquid culture.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.459-463
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• 1997

A bacterium DGUM 2011 has been selected from various samples of industrial wastewater and soil. Based on the morphological and physiological characteristics, the isolate DGUM 2011 was identified as Rhodococcus sp. and named as Rhodococcus sp. DGUM 2011. The optimal temperature and pH for the cell growth of Rhodococcus sp. DGUM 2011 were 37$\circ$C and 7.6, respectively. When phenol was added to the minimal media as a sole source of carbon and energy, the concentrations of maximum and optimum for cell growth was 0.10% and 0.08%, respectively. When 0.05% phenol was given in the minimal media, Rhodococcus sp. DGUM 2011 completely utilize it within 24 hrs. The isolate could utilize benzoic acid, p-hydroxybenzoate, p-cresol, tyrosine and phloroglucinol. The isolate possessed both catechol 1,2-dioxygenase and 2,3-dioxygenase activity. However, the activity of catechol 1,2-dioxygenase was much higher than that of 2,3-dioxygenase, which suggests that the isolate might degrade phenol via both ortho- and meta-cleavage, mainly via ortho-cleavage.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.969-973
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• 2006

Stimulatory effects of three different intermediate metabolites (1-hydroxy-2-naphthoate, salicylate, and catechol) as potential inducers on phenanthrene degradation were investigated using two different bacteria (Pseudomonas putida ATCC 17484 and Burkholderia cepacia PB12). The relative induction capacity was high in the sequence of 1-hydroxy-2-naphthoate, salicylate, and catechol in both strains. The highest of up to 12 times increase of the induction was obtained by the addition of 1-hydroxy-2-naphthoate in the strain PB12, compared with the control where no exogenous inducer was added. The induction capacity of the potential inducers was closely related with the number of oxygenations required per electron equivalents in one mole of the inducer.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.79-84
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• 2001

An aniline-utilizing microorganism identified as a species of Pseudomonas was isolated from soil contaminated highly with aniline and urea-herbicide. This strain was able to utilize aniline as the sole source of carbon and energy, and was shown to harbor a single large plasmid mediating the aniline assimilation. Subsequent plasmid-curing of this bacterium resulted in the abolishment of the aniline utilizing phenotype and the loss of catechol-C2,3O-oxygenase. The reestablishment of the plasmid, denoted pB10, in cured Pseudomonas sp. via filter surface mating, resulted in restoration of the aniline assimilation abilities and enzyme activity.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.245-249
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• 2007

Chloroanilines are widely used in the production of dyes, drugs and herbicides. Chloroanilines, however, are considered potential pollutants due to their toxic and recalcitrant properties to humans and other species. With the increase of necessity of bioremediation, this study was conducted to isolate the chloroanilines-degrading bacteria. A bacterium capable of growth on 3,4-dichloroaniline (DCA) was isolated by the 3,4-DCA-containing enrichment culture. The strain KB35B was identified as Pseudomonas sp. and also able to degrade several chloroanilines. The isolated strain showed high level of catechol 2,3-dioxygenase activity in the presence of 3,4-DCA. The activity of catecho1 2,3-dioxygenase was supposed to be ones of the important factors for 3,4-DCA degradation. The activity toward 4-methykatechol was 60.6% of that of catechol, while the activity toward 3-methylcatechol and 4-chlorocatechol were 27.0 and 13.5%, respectively.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.339-345
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• 2009

We evaluated the activity and abundance of the crude-oil-degrading bacterium Nocardia sp. H17-1 during bioremediation of oil-contaminated soil, using real-time PCR. The total petroleum hydrocarbon(TPH) degradation rate constants(k) of the soils treated with and without H17-1 were $0.103\;d^{-1}$ and $0.028\;d^{-1}$ respectively. The degradation rate constant was 3.6 times higher in the soil with H17-1 than in the soil without H17-1. In order to detect and quantify the Nocardia sp. H17-1 in soil samples, we quantified the genes encoding 16S ribosomal RNA(16S rRNA), alkane monooxygenase(alkB4), and catechol 2,3-dioxygenase(23CAT) with real-time PCR using SYBR green. The amounts of H17-1 16S rRNA and alkB4 detected increased rapidly up to 1,000-folds for the first 10 days, and then continued to increase only slightly or leveled off. However, the abundance of the 23CAT gene detected in H17-1-treated soil, where H17-1 had neither the 23CAT gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity, did not differ significantly from that of the untreated soil($\alpha$=0.05,p>0.22). These results indicated that H17-1 is a potential candidate for the bioaugmentation of alkane-contaminated soil. Overall, we evaluated the abundance and metabolic activity of the bioremediation strain H17-1 using real-time PCR, independent of cultivation.

• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.237-241
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• 1997

Two isolated strains, Comamonas testosteroni CPW301 and Arthrobacter ureafaciens CPR706, were able to use 4-chlorophenol (4-CP) as a sole carbon and energy source. CPW301 was found to degrade 4-CP via a meta-cleavage pathway in which the chloro-substituent was eliminated even when 4-chlorocatechol was cleaved by the catechol 2, 3-dioxygenase. In contrast, CPR706 removed chloride from 4-CP prior to the ring-fission reaction, producing hydroquinone as a transient intermediate during 4-CP degradation. CPR706 exhibited much higher tolerance for 4-CP than CPW301, which was indicated by the maximum degradable concentration and degradation rate.