• Journal of Life Science
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• v.9 no.5
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• pp.525-530
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• 1999

The isolation and identification of bacteria from freshwater plant root system were performed. Twenty four different strains were identified by microbiological identification system. Gram negative bacteria were isolated three times more than Gram positive ones. The ratio of rod and coccus was 11:1. The similarity above 0.5 was 37.5% from the tested samples in identification. Among these isolated bacteria, three dominant strains Pseudomonas cepacia JH10, Xanthomonas maltophilia JH12, Aeromonas salmonicida JH13 were selected to investigate biochemical properties and optimal growth conditions. These strains reached maximum growth after 24-36 hr of incubation in peptone broth, and their optimal temperature was 28$^{\circ}C$ and that of pH was between 7 and 8.

• Journal of Microbiology
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• v.33 no.3
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• pp.208-214
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• 1995

The effects of various 2, 4-D-degradative plasmids on the axenic growth patterns, the degradation phenotypes, and the competitiveness of different host bacteria were evaluated in liquid cultures; the organisms and plasmids used were Alcaligenes eutrophus JMP134/pJP4, Alcaligenes paradoxus/p2811, Pseudomonas pickettii/p712, pJP4, and p712 or p 2811 exhibited very different restriction fragment profiles in restriction endonuclease digests. These plasmids were transferred to the recipients (P. cepacia and Alcaligenes JMP228) at relatively high frequencies ranging from 8.9 $\times$ 10$^3$ to 1.6 $\times$ 10$^5$ per donar cell. In the axenic liquid cultures the fast-growing strains, such as P. pseudomallei/p745 and P. cepacia/pJP4, exhibited short lag periods, high specific growth rates, and high relative fitness coefficients, while the slow-growing strains, such as P. pickettii/p712 and A. paradoxus/p2811, had long lag periods, low specific growth rates, and low relative fitness coefficients. Depending on the type of plasmid containing the genes for the 2, 4-D pathway, some transconjugants exhibited intermediate grwoth patterns between the fast-growing strains and the slow-growing strains. The plasmid and plasmid-host interactions determined specific growth rate and lag time, respectively, which were shown to be principal determinants of competitiveness among the strains, but relative fitness coefficient derived from the axenic culture was not always predictive for the mixed culture condition.

• The Plant Pathology Journal
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• v.15 no.4
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• pp.217-222
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• 1999

Treatment with antagonistic rhizobactera Burkholderia cepacia strain N9523 or an inducer of resistance DL-$\beta$-amino-n-butyric acid (BABA) effectively inhibited Phytophthora capsici infection on pepper plants in artificially infested pots. Treatment with BABA alone at $1,000\mu\textrm{g}$/ml or together with B. cepacia in combination induced a strong protection from the Phytophthora disease in the greenhouse. In artificially infested field, protection of pepper plants against the Phytophthora epidemic by BABA treatment was maintained at a considerable level. In contrast, soil drench with the antagonist B. cepacia alone, or in combination with BABA did not suppress the Phytophthora epidemic in the field. Mortality of pepper plants caused by P. capsici infection was significantly reduced by treatment with the antagonist Pseudomonas aeruginosa strain 950923-29 and BABA (12-29% plants diseased) relative to the untreated control (41-91% plants diseased) in the naturally infested field. Treatment with the antagonist Ps. aeruginosa strain 950923-29 and BABA also resulted in high levels of protection against Phytophthora blight in pepper plants. In the plastic filmhouse test, the average percentage of plants diseased was significantly low relative to the naturally infested field. Treatment with the antagonist Ps. aeruginosa strain 950923-29 and BABA in combination was most effective in suppressing the Phytophthora disease in field and plastic filmhouse.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.26-32
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• 2000

Catechol 2,3-dioxygenase was purified from recombinant strain E. coli CNU312 carrying the tomB gene which was cloned from toluene-degrading Burkholderia cepacia G4. The purification of this enzyme was performed by acetone precipitation, Sephadex G-75 chromatography, electrophoresis and electro-elution. The molecular weight of native enzyme was about 140.4 kDa and its subunit was estimated to be 35 kDa by SDS-PAGE. It means that this enzyme consists of four identical subunits. This enzyme was specifically active to catechol, and$K_(m)$ value and $V_(max)$value of this enzyme were 372.6 $\mu$M and 39.27 U/mg. This enzyme was weakly active to 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol, but rarely active to 2,3-DHBP. The optimal pH and temperature of the enzyme were pH 8.0 and $40^{\circ}C$. The enzyme was inhibited by $Co^(2+)$, $Mn^(2+)$, $Zn^(2+)$, $Fe^(2+)$, $Fe^(3+)$, and $Cu^(2+)$ ions, and was inactivated by adding the reagents such as N-bromosuccinimide, and $\rho$-diazobenzene sulfonic acid. The activity of catechol 2,3-dioxygenase was not stabilized by 10% concentration of organic solvents such as acetone, ethanol, isopropyl alcohol, ethyl acetate, and acetic acid, and by reducing agents such as 2-mercaptoethanol, dithiothreitol, and ascorbic acid. The enzyme was inactivated by the oxidizing agent $H_(2)$$O_(2)$, and by chelators such as EDTA, and ο-phenanthroline.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.313-319
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• 2004

The distribution and characteristics of acidotolerant heterotrophic and naphthalene-degrading bacteria were investigated in two forest areas, one near Ulsan petrochemical industrial complex (Sunam) and the other in countryside (Daeam). Average values of soil pH at Sunam and Daeam were 3.8 and 4.6, respectively. When het­erotrophic and naphthalene-degrading bacteria were enumerated by most probable number (MPN) procedures at Sunam, the median values of heterotrophs growing at pH 7.0 and pH 4.0 were $5.3{\times}10^7\;and\;3.3{times}10^7$ MPN/g, whereas those of naphthalene-degraders were $5.6{\times}10^4\;and\;4.0{times}10^5$ MPN/g, respectively. While the medians of heterotrophs at Daeam were larger than those at Sunam, the concentrations of naphthalene-degraders were higher at Sunam compared to those at Daeam. From the MPN tubes and enrichment cultures, we obtained 17 isolates of naphthalene-degraders which were identified as Sphingomonas paucimobilis, Brevundimonas vesic­ularis, Burkholderia cepacia, Ralstonia pickettii, Pseudomanas fluorescens, and Chryseomonas luteola. Among them, 6 isolates showed higher naphthalene-degrading activity on minimal media of pH 4 compared to pH 7, whereas the extent of growth was not greater at pH 4 than at pH 7 when they were inoculated on nutrient-rich media. It is plausible that the pH may affect naphthalene-degrading activity of the isolates by changing fatty acid composition of bacterial membrane.

• KSBB Journal
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• v.6 no.4
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• pp.363-368
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• 1991

PVA degrading bacteria were isolated from water system, and identified as Pseudomonas cepacia and Pseudmonas pseudomallei, which were named as Pseudomonas sp. G5Y and Pseudomonas sp. PW. It was found out that those two kinds of bacteria have a symbiotic relationship to degrade PVA. For the mixed culture of these bacteria, the optimal conditions of pH, temperature, nitrogen source, and polymerization degree of PVA were found to be 7.5, $35^{\circ}C$, ammonium sulfate, and 500, respectively. Also, the growth of these bacteria was promoted by trace elements such as vitamin B1, B12, pyridoxine, and p-aminobenzoate, respectively. The specific growth rate of mixed bacteria was inhibited when the concentration of PVA was more than 20g/l. The substrate inhibition kinetics of the mixed culture was $${\mu}=\frac{0.065S}{2.56+S+(S^2/156}hr^{-1}$

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.109-114
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• 2006

Yam has been recognized as healthy food due to its various biological activities, such as anti-obesity, antimicrobial, anticancer and immuno-stimulation activities, and its consumption has been increased during last decades. In this study, to investigate low-temperature, long-term storage of yam and to develop processed yam products, yam-putrefactive psychrotrophic bacteria were isolated from rotted yam and identified based on BBL identification system, fatty acid analysis in cell membrane and 16S rDNA sequence analysis. The putrefaction activity of isolated thirteen bacteria was evaluated using yam-slices (NaOCl-treated, autoclaved yam and without treatment), and YAM-10 and YAM-12 were identified as major psychrotrophic putrefactive bacteria. Both YAM-10 (Pseudomonas cepacia) and YAM-12 (Pseudomonas rhodesiae) bacteria grew well at 4$\sim$12$^{\circ}C$ and showed strong activity of polymer degrading enzymes, especially amylase, carboxy methyl cellulase and xylanase, at 20$^{\circ}C$. But they failed to grow at acidic pH (<5) or alkaline pH (>10). Our results suggested that the control of psychrotrophic Pseudomonas sp. by pH change and inhibition of polymer degrading enzymes, such as amy-lase, are necessary to long-term storage of yam.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.57-63
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• 2003

Using fungal (Fusarium solani f. pisi) and bacterial (Pseudomonas mendocina) cutinases, the initial hydrolysis rate of p-nitrophenyl esters was systematically estimated for a wide range of enzyme and substrate concentrations using a 96-well microplate reader. Both cutinases exhibited a high substrate specificity; i.e. a high hydrolytic activity on p-nitrophenyl butyrate (PNB), yet extremely low activity on p-nitrophenyl palmitate (PNP). When compared to the hydrolysis of PNB and PNP by other hydrolases [lipases and esterases derived from different microbial sources, such as bacteria (Pseudomonas cepacia, Psedomonas furescens, Baciilus stearothermophilus), molds (Aspeillus niger, mucor miehei), and yeasts (Candida rugosa, Candida cylindracea)], the above substrate specificity would seem to be a unique characteristic of cutinases. Secondly, the hydrolytic activity of the cutinases on PNB appeared much faster than that of the other hydrolytic enzymes mentioned above. Furthermore, the current study proved that even when the cutinases were mixed with large amounts of other hydrolases (lipases or esterases), the Initial hydrolysis rate of PNB was determined only by the cutinase concentration for each PNB concentration. This property of cutinase activity would seem to result from a higher accessibility to the substrate PNB, compared with the other hydrolytic enzymes. Accordingly, these distinct properties of cutinases may be very useful in the rapid and easy isolation of various natural cutinases with different microbial sources, each of which may provide a novel industrial application with a specific enzymatic function.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.7
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• pp.1025-1035
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• 2014

This study assessed the effect of immobilized lipase on weak base styrene resin using polyethyleneimine (PEI) with cross-linking. Two procedures were used in this study. The first one, "mono-layer" lipase immobilization, involves washing PEI after adsorption. The second procedure, "multi-layer" lipase immobilization, has no washing before the cross-linking step. Treverlite XS-100200 (weak base styrene resin) was immersed with PEI solution (2.2 mg/mL). Lipase AH (from Burkholderia cepacia) was adsorbed onto the support coated with PEI before cross-linking with glutaraldehyde. Structured lipid was synthesized by immobilized lipase-catalyzed interesterification using canola oil, palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). Total fatty acid contents of triacylglycerol (TAG) in structured lipids were analyzed to investigate activity, properties, and reusability of immobilized lipases. Activities of immobilized lipases on the multi-layer and mono-layer increased at a high concentration (8 mg/mL) of lipase solution used for immobilization. The results show that immobilized lipase with the mono-layer method at pH 8.0 on resin had the highest total saturated fatty acid content (26.17 area%). Activity of immobilized lipase with the multi-layer method at pH 7.5 on support was lower than that of the mono-layer, but total saturated fatty acid content was 16.79 area% higher than that of lipase AH (15.01 area%).

• Applied Biological Chemistry
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• v.39 no.5
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• pp.403-408
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• 1996

Total bacterial community DNA, which was extracted from microcosm soil and field soil after 2,4-D amendments, was analyzed on Southern blots, using the tfdA gene probe derived from plasmid pJP4 and the Spa probe from Sphingomonas paucimobilis. Southern blot analyses with total bacterial DNA extracted from soils Inoculated with Pseudomonas cepacia/pJP4 revealed that DNA probe method could detect the 2,4-D degrading bacteria down to $10^5\;cells/g$ dry soil. In the microcosm experiment, there was a good correlation between 2,4-D degradation and banding patterns in hybridization analyses performed after each 2,4-D treatment using the two probes. When bacterial DNA extracted from microcosm soil was hybridized with the Spa probe, a change in the position of hybrid bands was observed over time in a Southern blot, suggesting that population change or possibly genetic rearrangement in 2,4-D degrading microbial populations occurred in this soil. With the Spa probe, one hybrid DNA band was persistently observed throughout the five 2,4-D additions. When bacterial DNA isolated from the field soil was probed with the tfdA and Spa, strong hybridization signal was observed in the 100 ppm-treated subplot, weak signal In the 10 ppm-treated subplot, and no significant signal in the 1 ppm-treated and control subplots. The data show that DNA probe analyses were capable of detecting and discriminating the indigenous 2,4-D degrading microbial populations in soil amended with 2,4-D under laboratory and field conditions.