• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.62-67
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• 1997

A potential biocontrol bacterium, YB-70 was isolated from a rhizosphere in suppressive soil and identified as a strain of Bacillus subtilis. In several biochemical and in vitro antibiosis tests on Fusarium solani with the culture filterates from B. subtilis YB-70, we found that antifungal mechanism of B. subtilis YB-70 was mediated by antibiotic substances produced from the bacterium. These antifungal substances were appeared to be hear-resistant, micromolecular, and ethy alcohol soluble. Antifungal agents produced by B. subtilis YB-70 showed strong inhibified against root-rotting fungi F. solani in in vivo pot test. An antifungal substance. YBS-1s, was purified from the culture broth of B. subtilis YB-70 by isoelectronic precipitation, silica gel column chromatography and Sephadex LH-20 column chromatography analysis by Fab-MASS, $^{1}$H-NMR, $^{13}$C-NMR, DEPT, and amino acid analyzer revealed that the YBS-1A was a peptide antibiotics of iturin class containing seven amino acids from five different groups, and the other(YBS-1B) was an analogue of iturin group composed of 11 amino acids with larher molecular weight of about 1, 500 dalton, which was lager than that of iturin A.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.26 no.4
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• pp.305-311
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• 1993

Marine organisms were investigated to identify the marine actinomycetes that produced noble bioactive compounds. Microorganism counts range from $2.1{\times}10^3\;to\;1.2{\times}10\;CFU/g$ of marine organisms. Actinomycetes constituted 0.01 to $0.5\%$ of culturable microbial community. We identified the marine actinomycetes that produced novel bioactive compounds. During the course of screening for bioactives from the marine microorganisms, we found that the strain in sponge had antimicrobial activities. From the morphological, cultural and various physiological characteristics, this strain was identified for Actinomycetes No. 101. The optimal compositions of culture medium for Actinomycetes No. 101 were starch 30g/l as carbon source, casamino acid 10g/l as nitrogen source. The optimal pH of medium and fermentation temperature were $6.5{\sim}7.0$ and $30^{\circ}C$, respectively. Fermentation has been conducted in the marine broth at $30^{\circ}C$ for 72 hour. The yield of fermentation got about 3g as dry weight(per liter of broth). The distribution of antimicrobial activity of Actinomycetes No. 101 was screened by paper disc. The extract of cultured cell and broth inhibited the growth of Staphylococcus aureus and Bacillus subtilis, but the inhibition action was week against yeast and mold.

• Microbiology and Biotechnology Letters
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• v.32 no.4
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• pp.312-316
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• 2004

In oder to select the powerful rhizophere-dorminatable biocontrol agent, we had isolated an indigenous antagonistic bacterium which produced antibiotic and siderophore from a disease suppressive local field soil of Gyungsan, Korea. And we could select the Pseudomosp. 4059 which can strongly antagonize against Fusarium oxysporum and Phytophthora capsici by two kinds of antifungal mechanism that can be caused by the antibiotic of Phenazin, a siderophore and a auxin like subThe selected strain was identified as Pseudomonas fluorescens (biotype A) 4059 by biochemical tests, API $\textregistered$ test, MicroLog TM system and 16S rDNA analysis. The selected antagonistic microorganism, Pseudomosp. 4059 had an antifungal mechanism of antifungal antibiotic and sidrophore. And we were confirmed the antagonistic activity of P fluorescens 4059 with in vitro antifungal test against Phytophthora capsici and in vivo by red-pepper.

• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.379-386
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• 1998

Deterioration of food is in general caused by the presence of microorganisms and chemical compounds of food itself. There exists antimicrobial compound in the food, however, addition of food antiseptics, additives, or physico-chemical processing is a common practice. The safety of artificial chemical antiseptics became a serious public concern, therefore, new natural antiseptic compounds are in need to be developed. We have isolated a new natural antifungal protein (KBS-B2) from Astragal seed through ammonium sulfate precipitation and column chromatography using FPLC Mono-S and Superose 12HR. The purified protein inhibited growth of Candida albicans, and spore germination of food spoiling fungi such as Aspergillus ochraceus, Penicillium expensum, P. digitatum and Botrytis cineria. Antifungal effect of the KBS-B2 protein could be directly assayed by bioautography overlaying the fungal spores on the electrophoresed acrylamide gel. The comparison of N-terminal amino acid sequences of the KBS-B2 with known antifungal protein revealed that had 50% homology to thaumatin and zeamatin like proteins.

• The Korean Journal of Mycology
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• v.20 no.3
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• pp.259-268
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• 1992

Fifty-three actinomycetes antagonistic to Phytophthora capsici and Magnaporthe grisea were isolated from rhizosphere soils in six pepper-growing areas and ashore soils. Thirty-two antagonistic actinomycetes, showing inhibition zone larger than 5 mm, were classified into 20 groups according to their colony morphology and color. The antagonistic activity against P. capsici greatly varied, which showed inhibition zone sizes in the ranges from 5.7 to 17.5 mm on V-8 juice agar and from 2.5 to 17 mm on tryptic soy agar. The antagonistic activity of some actinomycetes tested was remarkably different between the two test media. The antagonists showed a relatively broad antifungal spectrum, but their antibacterial activity was negligible, except for Pseudomonas solanacearum. Butanol extracts of culture filtrates from antagonistic actinomycetes inhibited mycelial growth of P. capsici and M. grisea, thereby confirming strongly antibiotic production in culture. Culture filtrates of some antagonistic actinomycetes completely inhibited Phytophthora blight in pepper plants.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.191-202
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• 1998

About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

• Applied Biological Chemistry
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• v.35 no.5
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• pp.395-398
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• 1992

In order to study the influence of a 6'-methyl group in ring C of griseofulvin ${\underline{(1)}}$ on the fungicidal activity, 6'-methyl group was replaced with a larger phenyl group as $({\pm})-6'-phenylgriseofulvin$ ${\underline{(3)}}$, $({\pm})-6'-epiphenylgriseofulvin$ ${\underline{(4)}}$, synthesized by a Diels-Alder cycloaddition. Their biological activities were examined against Botrytis allii (IFO 9430) and B. cinerea (AHU 9573). $({\pm})-6'-Phenylgriseofulvin $ ${\underline{(3)}}$ showed high activity in $25\;{\mu}g/disc$.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.227-232
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• 2008

Biological antagonists of Phytophthora capsici were isolated from soil in Gyeongbuk, Korea. Among the isolated bacteria, a Bacillus sp. was identified from l6S rDNA sequence analysis and named Bacillus sp. AM-651. Bacillus sp. AM-65l strain which can strongly a antifungal activity against Phytophthora capsici. Culture conditions for the maximum production of the antagonistic substance were optimized. The production of antibiotic were high on modified Davis mineral medium pH 7 at $30^{\circ}C$. The medium for highest production of the agonistic substance optimized. It is composed the best activity on glucose, $(NH_4)_2SO_4$ and $K_2HPO_4$ at 0.5%, 0.1%, and 0.7%, respectively. By time course of culture solution selected Bacillus sp. AM-65l, the culture solution after 48hrs had strongly growth inhibition rate against P. capsici. And culture solution of Bacillus sp. AM-651 was stable within a pH range $5{\sim}11$ and temperature range $4{\sim}70^{\circ}C$. Bacillus sp. AM-651 cultured broth shown fungal growth inhibitory activity against B. sorokiniana, B. cinerea, R. solani avove and beyond P. capsici and comparatively showed a high activity against C. gloeosporioides, B. dothidea, B. cinerea and F. graminearum by agar diffusion method.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.194-199
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• 2005

In oder to select an antifungal substance-producing antagonistic bacterium against Fusarium oxysporum casuing fusarium wilt on tomato, strain BK4 was isolated from local soil of Gyeoungbuk and was identified as Bacillus thuringiensis by 16s rDNA analysis, biochemical test, and Mcirolog TM 3.0 System. The antibiotic of B. thuringiensis BK 4 was highly produced at $30^{\circ}C$ in nutrient broth (pH 9.0). The crude antibiotic was even stable at $121^{\circ}C$ and more stable at slight alkalic condition than acid condition. It was also remained $50{\%}$ activity at pH 3.0. B. thuringiensis BK4 showed the inhibition of spore germination and the biocontrol ability against F. oxysporum causing fusarium wilt of tomato in vivo test. According to these results, B. thuringiensis BK4 was enough to use with a microbial agent for biocontrol against fusarium wilt.

• Journal of Applied Biological Chemistry
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• v.52 no.3
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• pp.116-120
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• 2009

We found out the new method of the consortium for the environmental friendly agriculture by 8 kinds of the selected antagonistic rhizobacteria. This research involved composition of mutual complementary consortium by each antagonistic function such as production of antibiotic, siderophore, antifungal cellulase and insoluble phosphate solubilization. The consortium No.11 among composed consortium candidates showed the most pepper growth promoting activity and Phytophthora blight suppression on the in vivo pot test of red-pepper plant. The consortium No. 11 is combination of PGPR Bacillus subtilis AH18 and Bacillus licheniformis K11. B. subtilis AH18 and B. licheniformis K11 both could produce the auxin, antifungal ${\beta}$-glucannase and siderophore. Also, they had mechanism for solubilization of insoluble phosphate. But, B. licheniformis K11 could produce the antibiotic of iturin which was able to inhibit Phytophthora capsici. We confirmed complementary noncompetitive mutualism between B. subtilis AH18 and B. licheniformis K11 of the consortium No.11. The results came out through treatment of two strains co-culture, treatment of individual culture and co-treatment of two individual cultures for the growth and Phytophthora blight suppression of red-pepper. The treatment of two strains co-culture didn't show a synergic effect in comparing sole treatment on the pepper growth promotion and Phytophthora blight suppression. But, when the pots were treated simultaneously with co-treatment of two individual cultures, an synergic effect was seen in the growth promotion of roots, stem, leaves and suppressed Phytophthora blight on red-pepper in vivo pot test.