• Journal of Microbiology and Biotechnology
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• v.29 no.9
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• pp.1375-1382
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• 2019

Phenylalanine hydroxylase from Chromobacterium violaceum (CvPAH) is a monomeric enzyme that converts phenylalanine to tyrosine. It shares high amino acid identity and similar structure with a subunit of human phenylalanine hydroxylase that is a tetramer, resulting in the latent application in medications. In this study, semirational design was applied to CvPAH to improve the catalytic ability based on molecular dynamics simulation analyses. Four N-terminal truncated variants and one single point variant were constructed and characterized. The D267P variant showed a 2.1-fold increased thermal stability compared to the wild type, but lower specific activity was noted compared with the wild type. The specific activity of all truncated variants was a greater than 25% increase compared to the wild type, and these variants showed similar or slightly decreased thermostability with the exception of the $N-{\Delta}9$ variant. Notably, the $N-{\Delta}9$ variant exhibited a 1.2-fold increased specific activity, a 1.3-fold increased thermostability and considerably increased catalytic activity under the neutral environment compared with the wild type. These properties of the $N-{\Delta}9$ variant could advance medical and pharmaceutical applications of CvPAH. Our findings indicate that the N-terminus might modulate substrate binding, and are directives for further modification and functional research of PAH and other enzymes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.4
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• pp.307-313
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• 1992

To study the seasonal variation of heterotrophic bacteria near the surrounding seawater of Porphyra forming area, samples were collected in the intertidal waters of Mokp'o of the Yellow Sea from February to December, 1990. Annual distribution of heterotrophic bacteria ranged from $7.5\times10^2\;to\;1.1\times10^5\;cfu/ml,$ annual distribution of physiological characteristic bacteria ranged from $5.0\times10\;to\;4.34\times10^4\;cfu/ml$ for proteolytic bacteria, from 0 to $1.35\times10^4\;cfu/ml$ for lipolytic bacteria and from 0 to $1.2\times10^4\;cfu/ml$ for amylolytic bacteria. Sixty-five percent of isolates from seawater were rods, and $76.7\%$ of isolates were Gram-negative. Most isolates were mesophiles and showed utilization of various carbon sources such as glucose, maltose, lactose, sucrose and arabinose. Most isolates also showed tolerance to a broad range of salt concentration. Dominant genus in seawater were Flavobacterium spp., in February, Moraxella spp., Acinetobacter spp. in March, Bacillus spp., Chromobacterium spp., Micrococcus spp., Vibrio spp. in July and Chromobacterium spp., Micrococcus spp., Bacillus spp. in November.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.17.2-17
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.65.2-65
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.66.1-66
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• 1999

• Korean Journal Plant Pathology
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• v.11 no.3
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• pp.258-264
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• 1995

Production and some properties of chitinolytic enzymes were investigated by 80% ammonium sulfate precipitates (crude enzymes) from culture supernatant of antagonistic bacteria, Chromobacterium violaceum strain C-61 and strain C-72, Aeromonas hydrophila, Aeromonas caviae, and Serratia marcescens. The maximum production of chitinase was obtained from the 3-day culture at 28$^{\circ}C$ in C. violaceum stains, the 6-day culture in S. marcescens, and the 2-day culture in A. hydrophila and A. caviae. In the optimum culture periods, chitinase activity of C. violaceum strains C-61 was 1.5, 5.5, 12.0 and 11.3 times higher than those of strain C-72, S. marcescens, A. hydrophila and A. caviae, respectively. However, N,N'-diacetylchitobiase activity was 3.2 times higher in S. marcescens than in C. violaceum strain C-61, and that of Aeromonas spp.was very low. On gels containing glycol chitin, chitinase of C. violaceum strains showed four isoforms of 54-, 52-, 50- and 37-kDa, whereas there were four isoforms of 58-, 52-, 48- and 38-kDa in S. arcescens, three isoforms of 70-, 58- and 54-kDa in A. hydrophila and six isoforms of 90-, 79-, 71-, 63-, 58- and 38-kDa in A. caviae. The chitinase of C. violaceum strain C-61 was most active at pH 7.0 and at 5$0^{\circ}C$ and was stable in ranges of pH 5.0~10.0 for 2 hours and of 0~5$0^{\circ}C$ for 30 min.

• KSBB Journal
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• v.13 no.6
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• pp.669-674
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• 1998

This study was carried out particularly focusing on he antitumor effect in combination with 5-fluorocytosine(5-FC), antifungal agent, and extracellular cytosine deaminase from Chromobacterium violaceum YK 391 against U-937, K-562 and SNU-C4 cells. While the addition of 10$\mu\textrm{g}$/100 ${\mu}\ell$ of anticancer agent, 5-fluorouracil(5-FU), to U-937, K-562 and SNU-C4 caused the decrease of proliferation 90%, 75% and 93% respectively, the addition of 20 $\mu\textrm{g}$/100 ${\mu}\ell$ of the extracellular cytosine deaminase and 10 $\mu\textrm{g}$/100 ${\mu}\ell$ of antifungal agent 5-FC caused the decrease of proliferation 80%, 70% and 90%, respectively. These results, therefore, reveal that this enzyme has the similar clinical effect for considering of adjuvant antitumor effect. From the above results, the treatment of 5-FC and the cytosine deaminase was very effective and showed the possibility to remove side effects which easily occur by the treatment of 5-FU only. An extracellular cytosine deaminase.

• Journal of Pharmacopuncture
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• v.25 no.1
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• pp.37-45
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• 2022

Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.110.3-111
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• 2003

The effect of biological control on the severity of hot pepper wilt disease was evaluated in the vinyl house with plants cultivated in the nursery soil containing chitin and chitinolytic microorganisms. The chitinolytic microorganisms, Trichoderma harzianum and Chromobacterium sp. strain C-61, were well survived in the nursery soil containing chitin. The hot pepper damping-off was markedly suppressed in the nursery soil containing chitin and chitinolytic microorganisms. The survival of chitinolytic microorganisms and suppression of damping-off were superior as the amounts of chitin added to the nursery soil increased, but growth of hot pepper was inhibited in the 10％ (w/w) chitin treatment. When the plants cultivated in the nursery soil containing 1％ chitin and chitinolytic microorganisms were transplanted in the vinyl house, the vegetative growth increased and the wilt disease was reduced as comparison with those of control.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.110.2-110
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• 2003

The effect of biological control on the severity of hot pepper wilt disease was evaluated in the vinyl house with plants cultivated in the nursery soil containing chitin and chitinolytic microorganisms. The chitinolytic microorganisms, Trichoderma harzianum and Chromobacterium sp. strain C-61, were well survived in the nursery soil containing chitin. The hot pepper damping-off was markedly suppressed in the nursery soil containing chitin and chitinolytic microorganisms. The survival of chitinolytic microorganisms and suppression of damping-off were superior as the amounts of chitin added to the nursery soil increased, but growth of hot pepper was inhibited in the 10％ (w/w) chitin treatment. When the plants cultivated in the nursery soil containing 1％ chitin and chitinolytic microorganisms were transplanted in the vinyl house, the vegetative growth increased and the wilt disease was reduced as comparison with those of control.