• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.921-928
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• 2002

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.368-374
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• 1988

Enzymological proprties of an extracellular cytosine deaminase from Bacellus polymyxa YL 38-3 were investigated. The extracellular enzyme was very stable, and optimum pH and temperature for the enzyme activity were found to be near pH 6.0 in 0.2M potassium phosphate buffer and at $30^{\circ}C$, respectively. 5-Fluorocytosine was converyed to 5-fluorouracil by the enzyme, but 5-methylcytosine was not to thymine by it. The enzyme activity was completely inhibited by some heavy metal ion such as 1mM of $Cd^{2-}$ and $Hg^{2+}$, and by 1mM of p-chloromercuribenzoate, respectively. The enzyme activity was inactivated about 75% by 1mM of o-phenanthroline and monoiodoacetate. But the enzyme activity was stimulated up to 200% by 1mM of 2-mercaptoethanol.

• Journal of Bio-Environment Control
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• v.21 no.2
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• pp.127-133
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• 2012

Inhibition effects on spore germination and mycelia growth for pepper anthracnose fungi (Collectricum acutatum) were investigated in vitro using eco-friendly agricultural materials as well as ecofriendly pesticides. The inhibition effect on mycelia growth of anthracnose fungi is the highest when the anthracnose mycelia were treated with a pesticide (commercial name: Koreayeok) that contains a mixture of Bacillus subtilis and Panibacillus polymyxa, resulting in 100% inhibition of the mycelia growth. Meanwhile, the range of 20~40% inhibition effects on the growth of anthracnose mycelia was observed with other commercial agricultural materials. The significant inhibition effects on spore formation of anthracnose fungus were shown in vitro with two water dispersible pesticides containing sulfur [BTB (100%) and SulfurStar (95.1%)], Koreayeok (95.0%), Borstar (99.0%) containing Bordeaux mixture, and Jihabudea-KM containing Psedomonas spp. (96.1%), respectively. Taken from these in vitro results of inhibiting of the spore germination and mycelia growth together, Koreayeok is the most effective on control of pepper anthracnose disease in vitro. In addition, two water dispersible pesticides containing sulfur (BTB and SulfurStar) and Borstar (99.0%) containing Bordeaux mixture are also significantly applicable to prevent pepper plants from anthracnose disease in vitro. It remains to be determined whether the selected eco-friendly agricultural materials in effective control of anthracnose in vitro can be used to control pepper anthracnose in field.

• Microbiology and Biotechnology Letters
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• v.4 no.4
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• pp.166-178
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• 1976

Taxonomic study of 11 strains of Bacillus coagulans and 14 strains of 13 spccies of Bacillus by deoxyribonucleic acid (DNA)-DNA hybridization were conducted. Among the 11 strains of B. coagulans, 6 were isolated from soil and the rest were the authentic strains obtained from American Type culture collection (ATCC) or the Institute for Fermentation, Osaka (IFO). All strains were examined to confirm as they are expected species of B. coagulans by the methods of Cordon et al. according to Bergey's Manual (8th ed.). The intraspecific DNA homology indexes among the 11 strains of B. coagulans using strain ATCC 7050 as the standard ($^3$H labeled input DNA) showed 76％ or, more, respectively. These findings accorded well with the results of the conventional taxonomic study according to the Bergey's Manual. The interspecific DNA homology indexes between B. coagulant strain ATCC 7050 and the type cultures of B. subtilis (168), B. licheniformis (IFO 12107), B. pumilus (IFO 12110), B. firmus (ATCC 14575), B. lentus (ATCC 10840), B. circulans (ATCC 4513), B. macelans (ATCC 8244), B. polymyxa (ATCC 842), B. sphaericus (ATCC 14577), B. brevis (ATCC 8246, IFO 12334), B. laterosporus (ATCC 64), and B. pantothenticus (ATCC 14576) respectively, showed 2 to 4％, while that of between B. coagulans ATCC 7050 and Escherichia coli K-12 was less than 1 ％.

• Journal of Microbiology
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• v.40 no.2
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• pp.104-108
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• 2002

The hazelnut leaf holer (Anoplus roboris Sufr,, Coleoptera: Curculionidae) is a devastating pest of hazelnut and oak trees. It causes approximately 20-30% economic damage to hazelnut production per year in Turkey. In the present study, in order to fond a more effective and safe biological control agent against A. roboris, we investigated the bacterial flora of the hazelnut leave holer, and tested them for insecticidal effects on it. According to morphological, physiological and biochemical tests bacterial flora were identified as Bacillus circulans (Ar1), Bacillus polymyxa (Ar2), Enterobacter sp. (Ar3) and Bacillus sphaelicus (Ar4). Insecticidal effects of bacterial isolates were performed on adult A. roboris. The highest insecticidal effect determined was 67% by B. sphaericus within eight days. The insecticidal effects of the other isolates (Ar1, Ar2 and Ar3) were determined as 33%, 47% and 47% within the same period, respectively.

• The Korean Journal of Mycology
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• v.20 no.2
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• pp.109-117
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• 1992

Chemotactic responses of five motile saprophytic and one phytopathogenic bacteria e.g. Agrobacterium radiobacter, Bacillus subtilis, B. polymyxa, Pseudomonas aeruginosa, P. fluorescens and Xanthomonas malvacearum towards exudate of Cochliobolus sativus conidia, Fusarium of oxysporum f. sp. ciceri chlamydospores, Macrophomina phaseolina sclerotia and Phytophthora drechsleri f. sp. cajani oospores were determined in vitro at different abiotic conditions. In general, a positive correlation (r=0.76 to 0.89; P=0.05) was observed between concentration of fungal exudates and attraction of bacterial cells. Similarly, a significant (P=0.05; r=+0.82 to 0.95) positive correlation was noticed between chemotactic response and incubation period. The chemotactic response of bacteria was greatly influenced by temperature and pH of the test fungal exudate. The optimum temperature for maximum chemotaxis was $25^{\circ}C$ for A. radiobacter, $30^{\circ}C$ for B. polymyxa, P. aerugionosa, P. fluorescens and X. malvacearum and $35^{\circ}C$ for B. subtilis. Fungal exudates maintained at pH 7 attracted maximum number of bacteria. The response of bacterial cells to exudates at pH 3 and 11 was not significantly (P=0.05) different than that to the buffer (control). Chemotaxis of bacteria was observed towards attractants (fungal propagules and their exudates) when they were kept apart and bridged with the capillaries filled with non-attractant (buffer) or attractant (exudate).

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.102-110
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• 1991

Alkalophilic Bacillus sp. YJ-451, which was isolated from soil at several area in Korea, produced a novel type of bacteriolytic enzyme (cell wall peptidoglycan hydrolase) extracellulary. The cell wall hydrolytic activity was identified as a clear zone on sodium dodecyl sulfate polyacrylamide gel electrophoresis containing 0.2% (w/v) cell wall of Bacillus sp. as substrate. This enzyme was successively purified 66 fold with 3.2% yield in culture broth by ammonium sulfate precipitation, CM-cellulose column chromatography, and gel filtration, followed by hydroxylapatite column chromatography. The molecular weight of the purified enzyme was estimated to be 27,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum pH and temperature for the activity of the enzyme were pH 10.0 and $50^{\circ}C$, respectively. The enzyme was stable between pH 5.0 and 10.0 and up to $40^{\circ}C$. Among the microorganisms used in this experiment the enzyme was active against most of gram negative strains and the genus Bacillus such as B. megaterium, B. licheniformis, B. circulans, B. pumilus, B. macerans, B. polymyxa. The release of dinitrophenylglutamic acid but not reducing group from cell wall peptidoglycan digested by the enzyme suggested that the enzyme is a kind of peptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan.

• The Korean Journal of Mycology
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• v.35 no.2
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• pp.115-120
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• 2007

Powdery mildew caused by Podosphaera fusca is one of the most important diseases in leafy lettuce (Lactuca sativa L.). Since the disease has been a threat to safe cultivation of leafy lettuce, its control methods have to develop to produce good quality of lettuce for farmer and consumer. Occurrence of lettuce powdery mildew is increasing more and more due to continuous cultivation of lettuce all through the year, non-removal of diseased plant parts of lettuce, spray of inadequate fungicides by mistaken acknowledge of lettuce powdery mildew for lettuce downy mildew, etc. The control effect of five fungicides against lettuce powdery mildew was examined in a plastic greenhouse located in Suwon. When fungicides were sprayed three times at 10 days-intervals in the early stage of occurrence of powdery mildew, the incidence of powdery mildew in the plants treated with kresoxim-methy SC, azoxystrobin SC, Ampelomyces quisqualis AQ94013 WP, Paenibacillus polymyxa AC-1 SC and Bacillus subtilis Y 1336 WP was 0.7%, 0.7%, 26.0%, 36.7% and 42.0%, respectively, whereas the incidence of non-treated control was 55.3% on eight days after final application. Phytotoxicity of five fungicides tested was not observed in lettuce seedling plants.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.36-47
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• 2016

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

• Applied Biological Chemistry
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• v.29 no.4
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• pp.422-428
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• 1986

The bacteria were isolated from ordinary Korean soy sauce and soybean Paste. After isolation, we investigated the bacteria which produces the characteristic flavor of ordinary Korean soy sauce and soybean paste, nothing the aroma, presence of amino acids, and free sugars. The results were obtained as follows. The bacteria isolated from ordinary Korean soy sauce and soybean paste were various Bacillus species. The isolated bacteria produced characteristic orders: 'Meju' order, the characteristic saline odor of ordinary Korean soy sauce or ordinary Korean soybean paste order, and enzymes: protease and amylase. The good characteristic saline odor of ordinary Korean soy sauce was produced by Bacillus licheniformis$(SSB_3)$. The good odor of ordinary Korean soybean paste was produced by Bacillus polymyxa$(SSB_4)$, Bacillus species$(SPB_1)$, Bacillus brevis$(SPC_2)$, and Bacillus lichniformis$(SPC_{2-1})$.