• Journal of Microbiology
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• v.35 no.4
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• pp.300-308
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• 1997

The catechol 2,3-dioxygenase (C23O) encoded by the Pseudomonas putida xylE gene was over-produced in Escherichia coli and purified to homogeneity. The activity of the C23O required the reduced form of the Fe(II) ion since the enzyme was highly susceptible to inactivation with hydrogen perocide but reactivated with the addition of ferrous sulfate in conjunction with ascorbic acid. The C23O activity was abolished by treatment with the chemical reagents, diethyl-pyrocarbonate (DEPC), tetranitromethane (TNM), and 1-cyclohexy1-3-(2-morpholinoethyl) car-bodiimidemetho-ρ-toluenesulfontate (CMC), which are modifying reagents of histidine, tyrosine and glutamic acid, respectively. These results suggest that histidine, tyrosine and glutamic acid residues may be good active sites for the enzyme activity. These amino acid residues are conserved residues may be good active sites for the enzyme activity. These amino acid residues are conserved residues among several extradion dioxygenases and have the chemical potential to serveas ligands for Fe(II) coordination. Analysis of random point mutants in the C23O gene derived by PCR technique revealed that the mutated positions of two mutants, T179S and S211R, were located near the conserved His165 amd Hos217 residues, respectively. This finding indicates that these two positions, along with the conserved histidine residues, are specially effective regions for the enzyme function.

• Journal of Microbiology
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• v.44 no.6
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• pp.632-640
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• 2006

In this study, the biodegradative activities of monocyclic aromatic compounds were determined from the multi-drug resistant (MDR) Acinetobacter baumannii, which were studied in the form of clinical isolates from a hospital in Korea. These bacteria were capable of biodegrading monocyclic aromatic compounds, such as benzoate and p-hydroxybenzoate. In order to determine which pathways are available for biodegradation in these stains, we conducted proteome analyses of benzoate, and p-hydroxybenzoate-cultured A. baumannii DU202, using 2-DE/MS analysis. As genome DB of A. baumannii was not yet available, MS/MS analysis or de novo sequencing methods were employed in the identification of induced proteins. Benzoate branch enzymes [catechol 1,2-dioxygenase (CatA) and benzoate dioxygenase $\alpha$ subunit (BenA)] of the $\beta$-ketoadipate pathway were identified under benzoate culture condition and p-hydroxybenzoate branch enzymes [protocatechuate 3,4-dioxygenas $\alpha$ subunit (PcaG) and 3-carboxy-cis,cis-muconate cycloisomerase (PcaR)] of the $\beta$-ketoadipate pathway were identified under p-hydroxybenzoate culture condition, respectively, thereby suggesting that strain DU202 utilized the $\beta$-ketoadipate pathway for the biodegradation of monocyclic aromatic compounds. The sequence analysis of two purified dioxygenases (CatA and PcaGH) indicated that CatA is closely associated with the CatA of Acinetobacter radiresistance, but PcaGH is only moderately associated with the PcaGH of Acinetobacter sp. ADPI. Interestingly, the fused form of PcaD and PcaC, carboxymuconolactone decarboxylase (PcaCD), was detected on benzoate-cultured A. baumannii DU202. These results indicate that A. baumannii DU202 exploits a different $\beta$-ketoadipate pathway from other Acinetobacter species.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.358-364
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• 1998

New microbial-control agents were prepared with B. thuringiensis strain NT0423 having unique properties which are different with other B. thuringiensis strains belonging to serotype 7[Kor. J. Appl. Entomol. 32: 426-432.]. Three B. thuringiensis formulations designated as BioBact 10%, 20% and 40%, were made with various combinations of adjuvants. These formulations showed good physical properties in wettability, suspensibility, particle size and adherence. In addition the result of SDS-PAGE analysis indicated that $\delta$-endotoxins remain stably in all formulations. Among the tested formulations, two wettable powder formulations, BioBact 20% and 40%, comprising 20% and 40% of B. thuringiensis technical powder showed the effective control against diamondback moth larvae (Plutella xylostella) in laboratory and field tests. Especially, when compared with commercial B. thuringiensis formulations (A and B commercial formulations) in field evaluation, BioBact 20% and 40% formulations showed equal activity up to 80% lethality and a good persistence effect which remain on leaves at least 7 days.

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.392-396
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• 1991

Mu d1(Ap lac) bacteriophage can be used to search for genes which are members of a common regulatory network without having to know the functions of the genes in advance. Aim was for obtaining the loci in the SOS network as well as temperature inducible loci. For this purpose, recA441 allele was used. This allele encodes a thermosensitive recA gene product; thus, the recA441 allele can be activated upon temperature upshift without by external DNA damage. Approximately 10, 000 colonies were screened, and then searched for the colonies which expressed .betha.-galactosidase higher level at 42.deg.C than at 30.deg.C. The strains identified fell into two dlasses; (i) ones in which the increased expression was $recA^{+}$ $lexA^{+}$ -dependent, that is, din(damage-inducible) genes which were due to the activation of recA441 allele and (ii) ones in which the increased expression was $recA^{+}$ $lexA^{+}$ -independent and only temperature-inducible, tin genes. Rough mapping position was obtained for these genes.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.999-1010
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• 2021

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment. They are highly toxigenic and carcinogenic. Probiotic bacteria isolated from fermented foods were tested to check their ability to degrade and/or detoxify PAHs. Five probiotic bacteria with distinct morphologies were isolated from a mixture of 26 fermented foods co-cultured with benzo(a)pyrene (BaP) containing Bushnell Haas minimal broth. Among them, B. velezensis (PMC10) significantly reduced the abundance of BaP in the broth. PMC10 completely degraded BaP presented at a lower concentration in broth culture. B. velezensis also showed a clear zone of degradation on a BaP-coated Bushnell Haas agar plate. Gene expression profiling showed significant increases of PAH ring-hydroxylating dioxygenases and 4-hydroxybenzoate 3-monooxygenase genes in B. velezensis in response to BaP treatment. In addtion, both live and heat-killed B. velezensis removed BaP and naphthalene (Nap) from phosphate buffer solution. Live B. velezensis did not show any cytotoxicity to macrophage or human dermal fibroblast cells. Live-cell and cell-free supernatant of B. velezensis showed potential anti-inflammatory effects. Cell-free supernatant and extract of B. velezensis also showed free radical scavenging effects. These results highlight the prospective ability of B. velezensis to biodegrade and remove toxic PAHs from the human body and suggest that the biodegradation of BaP might be regulated by ring-hydroxylating dioxygenase-initiated metabolic pathway.

• Weed & Turfgrass Science
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• v.5 no.4
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• pp.187-190
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• 2016

This study was conducted to determine phytotoxicity of domestic rice varieties and characteristic of sensitive gene involved in herbicide reaction of 4-HPPD (4-hydroxy phenylpyruvate dioxygenase) inhibiting herbicides. The five rice varieties were grown for 2 to 3 leaf stage on seedling trays and then transplanted into plastic pot: 4-Japonica type (Sangjubyeo, Sambaekbyeo, Sanduljinmi and Kumyoung) varieties and 1-Indica type variety (IR8). We trialled standard (14 g a.i. $10a^{-1}$) and double fold (28 g a.i. $10a^{-1}$) dose of benzobicyclon treatment at 10 days after transplanting in order to investigate phytoxicity. The Japonica-type Sangjubyeo showed no rice injury but Indica-type IR8 show 4-5 (standard) and 5-6 (double fold) phytoxicity levels. In spite of Japonica-type, Sanduljinmi and kumyoung showed 3-4 and 4-5 levels. Target resistant gene, Hydroxyphenylpyruvate dioxygenase inhibitor sensitive gene No.1 (HIS1) have been isolated from five domestic rice cultivars (Sangjubyeo, Sambaekbyeo, Sanduljinmi, Kumyoung and IR8). Results of the sequence through PCR, all five tested rice cultivars had HIS1 gene regardless to rice cultivars. And the difference between rice varieties from sequence of HIS1 were identified some variation in genes.

• Journal of Environmental Health Sciences
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• v.18 no.2
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• pp.92-101
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• 1992

Halogen substituted-phenol and analog phenol degrading strains were identified as Aeromonas, Moraxella, and Flavobacterium genus. Optimal degrading condition was generally 50~100 $\mu$M substituted-phenol as carbon source, $NH_4NO_3$ as nitrogen source, 30$\circ$C , and initial pH 7.2. $\rho$-Chlorophenol degrading strain of Aeromonas sp. C4 had biodegradability to the various substituted-phenols. Flavobacterium sp. M9 had substrate specificity to methyl substituted-function. Catechol was cleavaged by catechol 1, 2-dioxygenase in Aeromonas sp. C4, Moraxella sp. N7, and Flavobacterium sp. M9.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.275-275
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• 2002

Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as carbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

• BMB Reports
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• v.51 no.2
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• pp.57-58
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• 2018

An accurate diagnostic marker for detecting early-stage hepatocellular carcinoma (eHCC) is clinically important, since early detection of HCC remarkably improves patient survival. From the integrative analysis of the transcriptome and clinicopathologic data of human multi-stage HCC tissues, we were able to identify barrier-to-autointegration factor 1 (BANF1), procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 (PLOD3) and splicing factor 3b subunit 4 (SF3B4) as early HCC biomarkers which could be detected in precancerous lesions of HCC, with superior capabilities to diagnose eHCC compared to the currently popular HCC diagnostic biomarkers: GPC3, GS, and HSP70. We then showed that SF3B4 knockdown caused G1/S cell cycle arrest by recovering $p27^{kip1}$ and simultaneously suppressing cyclins, and CDKs in liver cancer cells. Notably, we demonstrated that aberrant SF3B4 overexpression altered the progress of splicing progress of the tumor suppressor gene, kruppel like factor 4 (KLF4), and resulted in non-functional skipped exon transcripts. This contributes to liver tumorigenesis via transcriptional inactivation of $p27^{kip1}$ and simultaneous activation of Slug genes. Our results suggest that SF3B4 indicates early-stage HCC in precancerous lesions, and also functions as an early-stage driver in the development of liver cancer.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.40 no.5
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• pp.282-287
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• 2007

Chloroanilines are aromatic amines used as intermediate products in the synthesis of herbicides, azo-dyes, and pharmaceuticals. 3,4-dichloroaniline (DCA) is the degradation product of some herbicides (diuron, propanil, and linuron) and of trichlorocarbanilide, a chemical used as an active agent in the cosmetic industry. The compound, however, is considered a potential pollutant due to its toxicity and recalcitrant property to humans and other species. With the increasing necessity for bioremediation, we sought to isolate bacteria that degraded 3,4-DCA. A bacterium capable of growth on 3,4-DCA as the sole carbon source was isolated from seaside sediment using a dilution method with a culture enriched in 3,4-DCA. The isolated strain, YM-7 was identified to be Pseudomonas sp. The isolated strain was also able to degrade other chloroaniline compounds. The isolated strain showed a high level of catechol 2,3-dioxygenase activity on exposure to 3,4-DCA, suggesting that this enzyme is an important factor in 3,4-DCA degradation. The activity toward 4-methylcatechol was 53.1% that of catechol, while the activity toward 3-methylcatechol, 4-chlorocatechol and 4,5-chlorocatechol was 18.1, 33.1, and 6.9%, respectively.