• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.603-613
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• 2019

This study was carried out to examine the antagonistic effect against phytopathogenic fungi of isolated strains from soil samples collected from Busan, Changwon, and Jeju Island: Botrytis cinerea, Colletotrichum acutatum, Corynespora cassiicola, Fusarium sp., Rhizoctonia solani, Phytophthora capsici, and Sclerotinia sclerotiorum. According to results of our studies, isolated strains showed an antagonistic effect against phytopathogenic fungi. Such an antagonistic effect against phytopathogenic fungi is seen due to the production of siderophores, antibiotic substances, and extracellular amylase, cellulase, protease, and xylanase enzyme activities. Extracellular enzymes produced by isolated strains were significant, given that they inhibited the growth of phytopathogenic fungi by causing bacteriolysis of the cell wall of plant pathogenic fungi. This is essential to break down the cell wall of plant pathogenic fungi and thus help plant growth by converting macromolecules, which cannot be used by the plant for growth, into small molecules. In addition, they are putative candidates as biological agents to promote plant growth and inhibit growth of phytopathogenic fungi through nitrogen fixation, indole-3-acetic acid production, siderophore production, and extracellular enzyme activity. Therefore, this study suggests the possibility of using Bacillus subtilis ANGa5, Bacillus aerius ANGa25, and Bacillus methylotrophicus ANGa27 as new biological agents, and it is considered that further studies are necessary to prove their effect as novel biological agents by standardization of formulation and optimization of selected effective microorganisms, determination of their preservation period, and crop cultivation tests.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.97-103
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• 2011

Two Gram positive bacterial strains, designated strain GC-1 and GC-4, were isolated from coastal seawater near Jeju Island in the Republic of Korea. The two strains were identified as members of the genus Bacillus, based on 16S rRNA gene sequencing and data for physiological characteristics analyses. A subtle difference in physiological and genotypical characteristics has led us to designate the strains GC-1 and GC-4. The strain GC-1 showed a 99.91% similarity in 16S rRNA gene sequencing with B. tequiliensis and B. subtilis subsp. inaquosorum and the strain GC-4 showed a 100% similarity in 16S rRNA gene sequencing with those of B. altitudinis, B. stratosphericus, and B. aerophilus. However, both strains exhibited different physiological and genotypical characteristics in many aspects from those of their phylogenetically closest neighbors listed above, which implies that genus Bacillus has diversified into various species during its evolutionary process.

• Journal of Life Science
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• v.14 no.5
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• pp.859-862
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• 2004

This report investigates antifungal activity and effects of growth promotion by biosurfactant produced from Bacillus sp. LP03 and TBM40-3 against fungus causing plants disease (Glay Mold-Botrytis cinerea). Antifugal activity against B. cinerea infeeted to leek (Allium tuberosum Rottler) exhibited better than antifungal agent farming drug (smilex, Dong bang agro., Seoul, Korea.) through the field test. After infected by plant's disease, the leaves growth and number are maintained under presenting biosurfactant produced strains. Especially, one of the strains, named Bacillus sp. LP03 showed strong antifungal activity on field studies.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.537-543
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• 2005

Subtilisin (EC 3.4.21.14) is the major extracellular alkaline serine protease of Bacillus species. Previously, we found that subtilisins did not migrate in the electrophoretic field in the Laemmili buffer system due to their high pI values (over 8.8); however, it formed a 'binding mode' at the top of the separating gel [5]. Utilizing this characteristic, four subtilisins from Bacillus sp. strains (e.g., B. subtilis 168, B. subtilis KCTC 1021, B. amyloliquefaciens KCTC 3002, and Bacillus sp. DJ-1 and DJ-4) were easily and quickly identified by an over-running electrophoretic technique with a miniscale culture supernatant (less than 20 ml) without any column chromatographic steps. Two subtilisins (DJ-l and a recombinant version) from Bacillus sp. DJ-l were characterized, and the enzymatic properties were determined by SDS-fibrin zymography and densitometric analysis. Based on this observation, the recombinant pro-subtilisin DJ-l showed the same 'binding mode,' similar to native subtilisin DJ-l. On the other hand, mature subtilisin DJ -1 without pro-peptide showed no enzymatic activity.

• Journal of Microbiology and Biotechnology
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• v.29 no.6
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• pp.887-896
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• 2019

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based pathogen identification relies on the ribosomal protein spectra provided in the proprietary database. Although these mass spectra can discern various pathogens at species level, the spectra-based method still has limitations in identifying closely-related microbial species. In this study, to overcome the limits of the current MALDI-TOF MS identification method using ribosomal protein spectra, we applied MALDI-TOF MS of low-mass profiling to the identification of two genetically related Bacillus species, the food-borne pathogen Bacillus cereus, and the insect pathogen Bacillus thuringiensis. The mass spectra of small molecules from 17 type strains of two bacilli were compared to the morphological, biochemical, and genetic identification methods of pathogens. The specific mass peaks in the low-mass range (m/z 500-3,000) successfully identified various closely-related strains belonging to these two reference species. The intensity profiles of the MALDI-TOF mass spectra clearly revealed the differences between the two genetically-related species at strain level. We suggest that small molecules with low molecular weight, 714.2 and 906.5 m/z can be potential mass biomarkers used for reliable identification of B. cereus and B. thuringiensis.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.40-44
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• 2003

Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. In order to develop a DNA marker specific for Bacillus anthracis and to discriminate this species from Bacillus cereus group, we applied the randomly amplified polymorphic DNA (RAPD)-PCR technique to a collection of 29 strains of the genus Bacillus, including 22 species of the B. cereus group. A 709-bp RAPD marker (KHT5) specific for B. anthracis was obtained from B. anthracis BAK. The PCR product of internal primer set from the KHT5 fragment distinguished B. anthracis from the other species of the B. cereus group.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.17-21
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• 1993

Eight strains of Bacillus thuringiensis were isolated from soil in Korea and characterized. The isolates were named HL-24, HL-25, HL-33, HL-34, HL-35, HL-38, HL-39, HL-40. Strains HL-24 and HL-25 produced irregular parasporal crystals, HL-33 and HL-35 produced bipyramidal crystals, and others were round form in their cells. The biochemical characteristics of the eight isolates were only minor different in specific characteristics to the known serotypes of Bacillus thuringiensis. The HL-25, HL-33 and HL-34 strains showed resistances to cephalothin, colistin and penicillin G, and HL-39 and HL-40 strains were resistant to penicilin G. The strains of HL-24, HL-25, HL-33 and HL-34 were toxic to Bombyx mori lavae and HL-24, HL-25, HL-38, HL-39 and HL-40 strains killed Culex pipiens 3rd instar larvae. The HL-24 and H25 strains showed lethal activity against two kinds of the larvae, however lethality against mosquito larvae was low.

• Journal of Life Science
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• v.23 no.2
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• pp.259-266
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• 2013

To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.1-5
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• 1995

Bacterial strains showing the fibrinolytic activity were screened from Chungkook-jang and Natto (Japanese traditional soy food). The strains isolated from Natto revealed a high level of fibrinolytic activity, wherease nearly half of the isolates from Chungkook-jang did not show the relevant activity. However, one strain isolated from Chungkook-jang showed the highest fibrinolytic activity (1.84 plasmin unit), and subsequently identified as Bacillus species. The fibrinolytic strain was designated as Bacillus sp. CK 11-4.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.279-284
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• 2007

Two bacterial strains isolated from soil (Bacillus subtilis strains: PS2 and RFO41) were evaluated to determine their promoting effect on the growth of tomato seedling under axonic and pot conditions. The production of phytohormone, such as indole-3-acetic acid, indole-3-butyric acid, gibberellin and zeatin by these two strains was investigated as possible mechanisms for plant growth stimulation. Both PS2 and RFO41 were shown to produce various phytohormones, and. the production of phytohormones was stimulated by the addition of peptone-rich brain heart broth medium. In addition, these bacteria exhibited high levels of phosphatase activity, which ranged from 2.18 to $2.7\;{\mu}\;{\rho}-nitrophenol/ml/hr$. PS2 and RFO41 were applied to the pot test for growth of tomato seed with phosphate. Root and shoot lengths of germinated tomato after 15 days were 45.5% and 36.5% longer than that of control in RFO41 treated samples, respectively. Baciller sp. PS2 and RFO41 may have a potential for biofertilizer in the agriculture.