• Korean Journal of Microbiology
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• v.39 no.1
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• pp.1-7
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• 2003

The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.224-229
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• 2003

A keratinolytic protease was purified from the culture medium of Pseudomonas sp. KP-364 by use of an assay of the hydrolysis of feather keratin. Membrane ultrafiltration and DEAE-cellulose ion-exchange resin and Sephadex G-150 gel chromatographies were used to purify the enzyme. The specific activity of the purified keratinolytic protease relative to that in the original medium was approximately 72-fold high. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Sephadex G-150 chromatography indicated that the purified keratinolytic protease is monomeric and has a molecular weight of 36 kDa. The optimal pH and temperature of the keratinolytic protease activity were 6.6 and 37 C, respectively, and the keratinolytic protease was relatively stable at pH value from 3.0 to 10.0 at 37 C for 1hour. The keratinolytic protease was inhibited by EDTA and EGTA, indicating that the keratinolytic protease was a kind of metalloprotease that require Li+ for cofactor.

• Journal of Microbiology
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• v.41 no.2
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• pp.95-101
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• 2003

The lipase gene (lipA) and its activator gene (lipB) of Pseudomonas sp. SW-3 were cloned and sequenced. The lipB was found to be present immediately downstream of lipA. The deduced amino acid sequences of lipA and lipB showed a high level of homology to those of other lipases belonging to the family I.1 of bacterial lipases. When lipA was expressed in Escherichia coli using T7 promoter, an active lipase was produced in cells carrying both lipA and lipB, but not in cells harboring only lipA. Recombinant lipase (rPSL) overproduced in an insoluble form was solubilized in the presence of 8 M urea, purified in a urea-denatured form and refolded by removing urea in the presence of the Ca$\^$2+/ ion. rPLS had maximum activity at pH 8.0 and 50$^{\circ}C$, was stable at pHs from 7.0 to 9.0 and below 50$^{\circ}C$, and showed the highest activity toward the p-nitrophenyl ester of palmitate (Cl6).

• Korean Journal of Microbiology
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• v.31 no.2
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• pp.129-134
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• 1993

The pchAB genes of Pseudomonas sp. DJ-12 produce the enzymes of 4-chlorobipheny] (4CB) dioxygenase and dihydrodiol dehydrogenase which act on the first and second steps in degradation of 4CB and biphenyl. The genes were cloned in E coli XLI-Blue. The pcbAB genes of about 2.2 kb in size were contained in the pCUlO1 hybrid plasmid in the cloned cell of CUIOI. The genes were found to have their own promoter and three restriction sites for HindlII. 2,3-dihydroxybiphenyl was detected by the resting cell assay, as the metabolite transformed from biphenyl by the cloned cell of CUIOI. This means that the pcbAB genes are well expressed in E. coli. But dechlorination was unlikely involved in the pchAB gene expression but was believed to occur by functioning on 4CBA produced after ring-cleavage of 4CB.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.6
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• pp.387-393
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• 2001

To use as a fundamental data for the sustainable agriculture, which is nowadays a major trend to keep the productivity and conserve the environment, 487 paddy soil samples were collected from 21 regions of the Gyeongnam Province and analyzed the chemical characteristics and microbial population of the soil. The microbial population densities were bacteria $298{\times}10^5$($4{\sim}3000{\times}10^5$ range), fungi $63{\times}10^3$($2{\sim}441{\times}10^3$ range), actinomycetes $19{\times}10^5$($0.2{\sim}1250{\times}10^5$ range), Bacillus sp. $53{\times}10^4$($4{\sim}890{\times}10^4$ range) and Pseudomonas sp. $198{\times}10^4CFU\;g^{-1}$($4{\sim}1724{\times}10^4CFU\;g^{-1}$ range), respectively. The microbial populations of the soil were in general higher in southern area than in the northern area of the Gyeongnam Province. The average ratio of bacteria/fungi population was 473. As soil clay content increased, the populations of aerobic bacteria, actinomycetes and Pseudomonas sp. were remarkably decreased. The ratio of aerobic bacteria and fungi was 1554 in sandy loam and clay loam 1144, while Bacillus sp./fungi ratio was 11 in clay loam and 10 in loam. On the topographical differences, aerobic bacteria and Bacillus sp./fungi ratio were the higher in coastal plains than any other areas. The microbial population densities from different soil types were generally lower in ill-drained paddy field than those of other paddy field. The content of $P_2O_5$, K, Ca, $NO_3-N$ and EC in soil were positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi, Bacillus sp. and Pseudomonas sp.. The soil organic matter and Mg content were also positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi and Bacillus sp.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.410-414
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• 1992

Numerical identification was carried out for an isolate of Streptomyces strain producing the extracellular p-lactamase inhibitor. Fifty taxonomic unit characters were tested and the data were analyzed numerically using the TAXON program. The isolate was identified to the major cluster 5 of Streptomyces and it was best matched to Streptomyces omiyaensis which is a synonym of Streptomyces exfoliatus. Therefore, it was concluded that the isolate was identified to be a strain (SMF 19) of Streptomyces exjbliatus.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.677-684
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• 2000

Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enhanced by the increase of agitation speed. Two forms of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0, and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both proteases were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were $50^{\circ}C$ and $45^{\circ}C$, respectively. About 56% of the original protease BK7-2 activity remained after being treated at $50^{\circ}C$ for 30 min but protease BK7-1 was rapidly inactivated at above $25^{\circ}C$. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.2
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• pp.204-210
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• 1998

The experiment was conducted to obtain the basic data required to characterize and improve rhizosphere environment of salt-accumulated greenhouse(SAG) soils by comparing the soil properties and the microbial flora of such soils to those of unprotected arable upland(UAU) soils. Soils were sampled from greenhouses and unprotected upland fields around the country. Microbial propulation, biomass C content and soil chemical properties were of interest. Population density of fluorescent Pseudomonas was high in UAU soils, while those of pathogenic Fusarium sp. and fluorescent Pseudomonas were low in SAG soils. With increasing soil organic matter(OM) content, the population densities of Bacillus sp., fluorescent Pseudomonas sp., Enterobacteriaceae, and microbial biomass C content increased. As soil electrical conductivity(EC) increased higher than $5.1dS\;m^{-1}$, the ratios of bacteria to fungi(B/F) and actinomycetes to fungi(A/F) and the population density of fluorescent Pseudomonas decreased remarkably. The soil pH was positively related to the population density of aerobic bacteria, while it was negatively related to that of fungi. The soil OM content was significantly correlated to the population densities of actinomycetes($r=0.226^*$). Bacillus sp.($r=0.334^{**}$), Enterobacteriaceae($r=0.276^*$), and the microbial biomass C content($R=0.439^{**}$). The population density of actinomycetes was also significantly correlated with soil exchangeable Ca($r=0.334^{**}$) and Mg($r=0.352^{**}$).

• Journal of Life Science
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• v.27 no.8
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• pp.930-936
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• 2017

The flocculating properties of bioflocculant Biopol32 produced by Pseudomonas sp. GP32 were investigated for application in industrial wastewater treatment. The major flocculating substance of bioflocculant Biopol32 was identified as polysaccharide. Many anionic flocculants need a counter ion to induce higher flocculating activity. The flocculating activity of bioflocculant Biopol32 was markedly increased by the addition of cationic ions ($Ca^{2+}$, $Al^{3+}$). The flocculating activity of bioflocculant Biopol32 was the most effective when 7.0 mM $CaCl_2{\cdot}2H_2O$ as coflocculant was added. The flocculating activity on the effect of pH and the temperature of the bioflocculant Biopol32 was compared with anionic commercial flocculant (polyacrylamide) and bioflocculant (zooglan from Zoogloea ramigera). In kaolin suspension, the highest flocculating activity was obtained at the bioflocculant Biopol32 concentration of 1.5 mg/l. A high flocculating activity was observed in the pH range of 5.0 to 8.0. The flocculating activity of bioflocculant Biopol32 was sustained up to $60^{\circ}C$, but decreased rapidly at over $70^{\circ}C$. In the batch culture, the charge density of bioflocculant Biopol32 was compared with flocculating activity. The larger the anionic charge density and apparent viscosity of bioflocculant Biopol32, the higher the flocculating activity. Therefore, we confirmed that the flocculating activity and apparent viscosity of bioflocculant Biopol32 was closely related to the charge density of bioflocculant Biopol32.

• KSBB Journal
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• v.10 no.5
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• pp.582-588
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• 1995

A marine microorganism producing biosurfactant was isolated from the oil polluted coast of Chung-Mu in Korea, and was identified as Pseudomonas sp.. It produced the biosurfactanl and its optimum culture conditions for pH and salt concentration were 8.0 and 3.0%, respectively. The productivity of biosurfactant from this strain was affected by the nitrogen source used. For the oil resolvability of the biosurfactant, the residual oil in the culture broth with 2% Kuwait crude oil at each time of 48, 96, and 132hr was investigated by gas chromatography. As result of this experiment, it was verified that the biosurfactant acted on C10-C14, of Kuwait crude oil and so the oil was decomposed. The biosurfactant isolated from the supernatant was purified by adsorption to Amberliter XAD-7 and followed by gel chromatography (Sephadex G-100) and HPLC. The purified biosurfactant showed a high value of emulsifying activity at $40^{\circ}C$ and the emulsifying stability was maintained at the temperature range of $30^{\circ}C$∼$60^{\circ}C$. The purified biosurfactant reduced the interfacial tension of Kuwait crude oil remarkably and showed improved dispersing ability compared to those of commercial surfactants such as Tween 80, Tween 60 and SDS.