• Korean Journal of Microbiology
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• v.32 no.4
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• pp.285-290
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• 1994

About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.212-219
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• 2016

Forty-nine pigments were extracted from the collections of 106 pigment producing bacteria from the plant rhizosphere soil. Antibacterial activity test was performed in the subjects of the extracted pigments with plant pathogenic bacteria including Xanthomonas axonopodis and Xanthomonas campestris, and with plant pathogenic fungi including Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum. The yellow pigment by Chryseobacterium sp. RBR9 and the red pigment by of Methylobacterium sp. RI13 showed the antibacterial activities against Xanthomonas axonopodis and Xanthomonas campestris. The violet pigment by Collimonas sp. DEC-B5 showed the antibacterial activity as well as the antifungal activities against Botrytis cinerea and Fusarium oxysporum. Especially, the violet pigment inhibited the growth of Botrytis cinerea more than 65% at MIC $20{\mu}M$. Upon the HPLC analysis result for the isolation of pigment with antifungal activity, violacein (91.6%) and deoxyviolacein (8.4%) were isolated for the pigment by Collimonas sp. DEC-B5. The production amount of the pigment was increased more than 10 times higher when D-mannitol 1.5% and yeast extract 0.2% were added as the nitrogen source to SCB medium. This study suggests that produced violacein by Collimonas sp. DEC-B5 will be effective to control strawberry gray-mold rot fungi by its preventive activity.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.326-332
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• 2008

Various bacteria were isolated from Korean soil samples based on their capability inhibiting the growth of MRSA strains. Among them, strain YK5 with the highest activity was a Gram positive sporulative bacillus with motility. It did not produce indole and no acid was formed from mannitol by the bacterium. The 16S rRNA sequence of the strain showed $95{\sim}98%$ homology with those of Paenibacillus spp.. The bacterial isolate shared the highest homology with that of P. elgii (98%), but was named as Paenibacillus incheonensis YK5 due to differences in physiological properties. Butanol extract of the P. incheonensis YK5 culture grown in SST medium at $37^{\circ}C$ for 96 hr showed a broad antimicrobial activity against Gram-positive (MRSA and Streptococcus pneumoniae) and negative (Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Escherichia coli, Klebsiella pneumoniae) pathogenic bacteria and fungi (Cryptococcus neoformans and Trichophyton). The antimicrobial activity in the crude extract was stable in a broad range of temperature and pH, $20{\sim}100^{\circ}C$ and $3.0{\sim}6.0$, respectively. Therefore, the antimicrobial activity of P. incheonesis YK5 had potential as a novel antibiotics for pathogens including MRSA.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.5
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• pp.612-617
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• 2009

Fresh vegetable juice is a non-heat treated product and the only step to reduce microbial growth is washing. Therefore, the materials for fresh vegetable juice including Angelica keiskei, Brassica loeracea var. acephala, and Daucus carota L. were treated by ozone after the first washing process and investigated for microbial and chemical changes. The number of the total aerobic bacteria in materials after selection step were $8.2{\times}10^5{\sim}5.0{\times}10^6\;CFU/g$, which was a higher contamination level than the limit of Korea food code ($10^5\;CFU/g$). However, after the 1st washing process and ozone treatment, the total aerobic bacterial number was reduced to $4.7{\times}10^4{\sim}6.7{\times}10^4\;CFU/g$, which showed 2 log microbial reduction. After the 2nd washing step followed by ozone treatment, there was no difference in microbial number. The number of colifroms in the materials of fresh vegetable juice were $8.0{\times}10^3{\sim}3.5{\times}10^3\;CFU/g$ initially but showed $1.5{\times}10^2{\sim}3.0{\times}10^2\;CFU/g$ after the ozone treatment (1 log reduction). On the other hand, there was no changes in the contents of ascorbic acid, flavonoids, polyphenols, minerals (cadmium and lead) during all processes. In addition, no color changes were observed during washing process. Therefore, ozone treatment in the materials of fresh vegetable juice decreased the microbial numbers. Also, chemical characteristics of ozone treated sample were not different when compared with control.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.657-663
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• 2005

Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

• The Korean Journal of Pesticide Science
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• v.19 no.1
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• pp.54-63
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• 2015

Streptomyces spp. isolated from turfgrass rhizosphere and tested for their response to large-patch control fungicides. The tested fungicides were actually used in golf course or turfgrass cultivation to prevent large-patch disease. Tolerance to 3 triazole group of the strains was the highest to the PR fungicide, and following the SR fungicide, whereas the isolated strains were no tolerance to HR fungicide. Tolerances to three kind of Strobilurin group were similar for the all of the tested Streptomyces spp.. Growth and sporulation of the all strain was normal in CB and AP fungicide treatments. However no spore formulated in double concentration. Strains, tolerance to acetanilide fungicides, appeared that KT fungicide tolerance was higher than MK fungicide. The selected strains showed strong tolerance against AT fungicide but have no tolerance to ATR fungicides. In conclusion, the bacterial strains showed tolerance against 1 carbamate, 1 organophosphate and 1 cyanopyrrole group, while have no tolerance against two mixture formulations (1 Quinone + Strobilurin and 1 Imidazole + Triazole).

• Applied Biological Chemistry
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• v.38 no.3
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• pp.191-195
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• 1995

Production of a high viscosity exoploysaccharide, methylan, by Methylobacterium organophilum from methanol was carried out in fed-batch cultures and the rheological properties of methylan fermentation broth were studied. Bacterial biomass showed little influence on viscosity, but the accumulation of methylan caused the increase of viscosity. With proceeding fermention, the viscosity at the same concentration of methylan was significantly increased and methylan solution showed slightly higher pseudoplasticity. The composition changes of methylan were investigated at various fermentation times. Contents of total sugar, reducing sugar and methylan were decreased but contents of acids(pyruvic acid, uronic acid and acetic acid) were increased with the culture time. It was considered that the increased content of acids resulted in the increase of the hyrodynamic domain in the solution due to charge repulsion. Consequently, the solution viscosity increased in propotion to the acids contents of methylan. Cell growth and methylan production were severely decreased by the limitation of dissolved oxygen. However, the cellular activity for methylan production was almost constant regardless of the level of dissolved oxygen. As a result, the high speed of agitation increased the methylan production, the specific production rate of methylan, and the methylan yield of the cell.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.190-195
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• 2000

Tolaasin is a peptide toxin produced by Pseudomonas tolaasii and causes a brown blotch disease forming brown, slightly sunken spots and blotches on the cultivated mushrooms. It is a lipodepsipeptide consisting of 18 amino acids and its molecular mass is 1,985 Da. It forms a pore in plasma membranes, resulting in the disruption of membranes of fungal, bacterial, plant, and animal cells as well as mushroom tissue. In order to measure the toxicity of tolaasin, erythrocytes of blood were used to evaluate the tolaasin-induced hemolysis. Hemolytic activity of tolaasin was measured by observing the absorbance change either at 420 nm, representing the release of hemoglobins from red blood cells(RBCs), or at 600 nm, representing the density of residual cells. The hemolytic activity of culture-extract of P. tolaasii increased at early-stationary phase of growth and was maximal at late stationary phase. The hemolytic activity of tolaasin appeared high in the RBCs of dog and rat. The RBCs of rabbit and hen were less susceptible to tolaasin. The effects of various cations were also measured. $Cd^{2+}$ and $La^{3+}$. as well as $Zn^{2+}$ appeared inhibitory to the tolaasin-induced hemolysis. The effects of various anions on tolaasin-induced hemolysis were measured and carbonate showed the greatest inhibition to the hemolysis. However, phosphate stimulated the tolaasin-induced hemolysis and no effects were observed by chloride and nitrate.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.545-550
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• 2002

A hemolysin producing bacterial strain which belong to Vibrio species was isolated from the Kum River estuary. In the process of identification, the strain did not show characteristics of known Vibrio species; thus, the strain was designated as Vibrio sp, E10 (V. kunsan) tentatively and further identification study was carried out by comparing its bacteriological characteristics. Morphologically Vibrio sp, E10 was comma shaped rod with a polar flagellium. Clear hemolysis zones were observed with the strain against human and sheep blood agar. Hemollytic toxicity was confirmed by strong vascular Permeability and fatal toxicity against mouse was also observed. Therefore the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. E10 were ranged salinity of 0$\~$$4.5\%$, pH of 6.2$\~$9.2, temperature of 14$\~$42$^{\circ}C$, respectively, 16S rDNA partial sequence of Vibrio sp, E10 showed $99\%$ homology with dozens of V. cholerae species including V, cholerae El Tor N16961 and V, snmisnfus ATCC 33653T. This strain belonged to Proteobacteria; gamma subdivision; Vibrionacea: Vibrio. But, among knorn Vibrio species no identical styains were found when using automatic bacteria identification system ($MicroLog^{TM}$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp, E10 showed 18 and 11 different responses as compared to V. mimicus and V, cholerae, respectively.

• Research in Plant Disease
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• v.18 no.2
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• pp.109-116
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• 2012

A bacterial strain antagonistic to some fungal phytopathogens was isolated from the stem of a Persimmon tree in Yeongam, Korea. This bacterium was identified as Bacillus subtilis by 16S rRNA gene sequencing and designated as B. subtilis GDYA-1. In in vivo experiment, the fermentation broth exhibited antifungal activities against Magnaporthe oryzae on rice plants, Phytophthora infestans on tomato plants, and Puccinia recondita on wheat plants. We isolated one antifungal compound and its chemical structure was determined by mass and $^1H$-NMR spectral data. The antifungal substance was identified as benzoic acid. It inhibited mycelial growth of M. oryzae, Rhizoctonia solani, Sclerotinia sclerotiorum, and P. capsici with minimum inhibition concentration (MIC) values, ranging from 62.5 to 125 ${\mu}g/ml$. Moreover, the substance effectively suppressed Phytophthora blight of red pepper caused by P. capsici in a pot experiment. To the author's knowledge, this is the first report on the antifungal activity of benzoic acid against phytopathogenic fungi. Benzoic acid and B. subtilis GDYA-1 may contribute to environmental-friendly protect crops from phytopathogenic fungi.