• Korean Journal of Microbiology
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• v.36 no.4
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• pp.259-266
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• 2000

Spheroplast cell fusions were performed with Flavimonas oryzihabitans degrading aniline and Pseudomonas sp. degrading 4-chlorobiphenyl to develope the new fusant degrading aniline and 4-CBP and its characters were investigated. F. oryzihabitans was induced to antibiotic marker ($Cm^r$ by NTG treatment for the fusants selection. The results of spheroplast formation and regeneration frequencies of the strains treated with lysozyme-EDTA were 99% and 5.0~6.6%, respectively. Fusion products were treated with 40% (v/v) PEG 6000 and fusion frequency was $3.16{\times}10^{-4} $. The DNA content of fusant, F22 was approximately 2-fold compared with parents. The fusant was stable, and showed the mixed biochemical characteristics of the parent strains. F22 was similar to parent for cell growth pattern and degrading capacity on 5 mM aniline but cell growth rate of F22 was 1.5-fold higher than that of the parent on 10mM aniline. However 4-CBP degrading ability of F22 was slightly lower than that of parental strain.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.263-270
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• 1992

For the selection of powerful antagonistic bacterium for biological control of soil borne Eminia carotovora subsp. carotovora causing rot of vegetable, excellent strains (S4, S14, 565) were selected from 1,196 strains of bacteria which were isolated from rhizosphere in vegetable root rot-suppresive soil. Strains were identified to be Pseudomonas species with Api 20NE kit. Antagonistic substance was produced in 523 synthetic broth medium at pH 7~8 and $30^{\circ}C$ during 3 days culture. The substance was stable in the pH range of 6 to 9. When the basal medium was supplemented with mannitol and sorbitol as carbon source and calcium chloride as metal salt, the production of the inhibitory substance was increased. The inhibitory acitivity was increased by the addition of fertilizer in soil. The isolated strains were resistant to the agricultural chemical such as benomyl and fosethyl-Al-folpet, and the antibiotics such as penicillin and lincomycin. We had found that Pseudomonas sp. S14 strain had a single plasmid. After treated with acridin orange for curing, we confirmed the existence of antagonistic gene in the chromosomal DNA.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.275-281
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• 2004

Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quin­oline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2­dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, puri­fication fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-meth­ylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.

• Korean Journal of Food Science and Technology
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• v.45 no.4
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• pp.515-520
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• 2013

Bacterial communities derived from cheonggukjang and raw rice straw collected from a Mireuksan farm and a Heungup cheonggukjang in Gangwon province were investigated using both culture-based method and denaturing gradient gel electrophoresis (DGGE) analysis. Pure cultures, which were isolated from raw rice straw and cheonggukjang and cultured on tryptic soy agar plates (53-76 colonies per plate), were identified by analysis of 16S rRNA sequences. The traditional culture-based method and analysis of PCR-amplified 16S rRNA by DGGE revealed that for samples collected from the Mireuksan farm, Pantoea agglomerans and Bacillus subtilis were the predominant species in the raw rice-straw and cheonggukjang, respectively. For samples collected from the Heungup cheonggukjang, Bacillus amyloliquefaciens was the predominant species in both raw rice straw and cheonggukjang. Other microorganisms, including members of Pantoea, Bacillus, Enterococcus, Enterobacter, Pseudomonas, Rhodococcus, and Acinetobacter, were also present in the raw rice-straw and cheonggukjang, as were bacteria that could not be cultured.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.475-480
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• 1989

The extracellular polysaccharide was isolated from the culture of Ps. delafieldii and its rheological property was evaluated. The aqueous solution was extremely viscous and shows pseudoplastic behaviour. The flow behaviour index and apparent viscosity of 1 ％solution were 0.09 and 1169 mPa·s. The solution was stable over pH change but did not have thermal stability. The activation energy of flow of 1 ％ solution was 4.44 kcal/mole. The concentration dependency could be expressed double logarithmically.

• Microbiology and Biotechnology Letters
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• v.16 no.5
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• pp.421-426
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• 1988

Twenty-nine bacterial isolates capable of growing on aniline as n sole source of carbon and nitrogen were obtained. Ten of these isolates were identified. Nine isolates were Identified as Pseudomonas spp. and one was Acinetobacter sp.. Five strains among 29 isolates had one or several plasmids. Four of these five strains utilized aniline through meta pathway and one through ortho pathway. Pseudomonas acidovorans 4A1 which utilized aniline through meta pathway harbored a plasmid of ca. 230 kilobases shown to be involved in aniline metabolism.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.67-73
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• 2002

The cellular responses of soil-borne bacterium, Pseudomonas sp. HK-6 to explosive 2,4,6-trinitrotoluene (TNT) were examined. Two stress shock proteins (SSPs), approximately 70-kDa DnaK and a 60-kDa GroEL were found in HK-6 cells in response to TNT. Analyses of SDS-PAGE and Western blot using anti-DnaK and GroEL revealed that SSPs were induced in HK-6 cells exposed to 0.5 M of TNT far 6-12 hrs. The maximum induction of proteins was achieved at 8-hr incubation point after HK-6 cells'exposure to TNT. Similar SSPs were found to be induced in HK-6 cells by heat shock (shift of temperature, from $30^{\circ}C$ to $42^{\circ}C$) or cold shock (shift of temperature,$30^{\circ}C$ to $4^{\circ}C$).2D-PAGE of soluble protein tractions from the culture of Pseudomonas sp. HX-6 exposed to TNT demonstrated that approximately 450 spots were observed on the silver stained gels ranging from pH 3 to pH 10. Among them, 12 spots significantly induced and expressed in response to TNT were selected and analyzed. Approximately 60-kDa protein, which was assumed highly expressed on the gel, was used for amino acid sequencing. N-terminal microsequencing with in-gel digestion showed that N-terminal sequence of the TNT-induced protein, <$^1XXAKDVKFGDSARKKML^17$, shared extensive similarity with $^1XXAKDVKFGDSARKKML^17$, N-terminal sequence of (P48216) GroEL of Pseudomonas putida.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.72-77
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• 2001

Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.343-353
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• 2018

Our previous research demonstrated the essential role of the xenB gene in stress response to RDX by using Pseudomonas sp. HK-6 xenB knockout. We have extended this work to examine the cellular responses and altered proteomic profiles of the HK-6 xenA knockout mutant under RDX stress. The xenA mutant degraded RDX about 2-fold more slowly and its growth and survival rates were several-fold lower than the wild-type HK-6 strain. SEM revealed more severe morphological damages on the surface of the xenA mutant cells under RDX stress. The wild-type cells expressed proportionally-increased two stress shock proteins, DnaK and GroEL from the initial incubation time point or the relatively low RDX concentrations, but slightly less expressed at prolonged incubation period or higher RDX. However the xenA mutant did not produced DnaK and GroEL as RDX concentrations were gradually increased. The wild-type cells well maintained transcription levels of dnaA and groEL under increased RDX stress while those in the xenA mutant were decreased and eventually disappeared. The altered proteome profiles of xenA mutant cells under RDX stress also observed so that the 27 down-regulated plus the 3 up-regulated expression proteins were detected in 2-DE PAGE. These all results indicated that the intact xenA gene is necessary for maintaining cell integrity under the xenobiotic stress as well as performing an efficient RDX degradation process.

• KSBB Journal
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• v.14 no.2
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• pp.153-159
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• 1999

Fifteen microbes have been isolated from Jangja pond in Kuri-Si, Kyeonggi-Do. Among them, two strains showed excellent COD removal from wastewater, which were named Pseudomonas sp. BLP2052 and Flavobacterium sp. BLP20515, respectively. Optimal pH and temperature for the cell growth were 7.0 and $30^{\circ}C$ for both strains. Pseudomonas sp. BLP2052 and Flavobacterium sp. BLP20515 was applied to the reactor to treat wastewater and 66.0% and 65.7% COD (chemical oxygen demand) removal was achieved, respectively. Comparing these results to the case of applying mixed microbes present in Jangja pond, COD removal rate was 15% less. But when adding the selected microbes to the wastewater containing mixed microbes, COD removal rate increased by 5%. After 84 hour operation, we achieved 85.6% COD removal. When inhibitors were added less than 100 ppm, during the microbial wastewater treatment, Fe, Zn, Al, phenol and Cr influenced microbial activity more deterioratively in order. In the case of over 300 pm, Cr, Fe, Zn, Al and phenol showed severe deteriorative effect in order.