• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.157-167
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• 2000

A thermostable chitosanase gene from the isolated strain, Bacillus sp. CK4, was cloned, and its complete DNA sequence was determined. The thermostable chitosanase gene was composed of an 822-bp open reading frame which encodes a protein of 242 amino acids and a signal peptide corresponding to a 30 kDa enzyme in size. The deduced amino acid sequence of the chitosanase from Bacillus sp. CK4 exhibits 76.6%, 15.3%, and 14.2% similarities to those from Bacillus subtilis, Bacillus ehemensis, and Bacillus circulans, respectively. C-terminal homology analysis shows that Bacillus sp. CK4 belongs to the Cluster III group with Bacillus subtilis. The size of the gene was similar to that of a mesophile, Bacillus subtilis showing a higher preference for codons ending in G or C. The functional importance of a conserved region in a novel chitosanase from Bacillus sp. CK4 was investigated. Each of the three carboxylic amino acid residues were changed to E50D/Q, E62D/Q, and D66N/E by site-directed mutagenesis. The D66N/E mutants enzymes had remarkably decreased kinetic parameters such as $V_{max}$ and k$\sub$cat/, indicating that the Asp-66 residue was essential for catalysis. The thermostable chitosanase contains three cysteine residues at position 49, 72, and 211. Titration of the Cys residues with DTNB showed that none of them were involved in disulfide bond. The C49S and C72S mutant enzymes were as stable to thermal inactivation and denaturating agents as the wild-type enzyme. However the half-life of the C211S mutant enzyme was less than 60 min at 80$^{\circ}C$, while that of the wild type enzyme was about 90 min. Moreover, the residual activity of C211S was substantially decreased by 8 M urea, and fully lost catalytic activity by 40% ethanol. These results show that the substitution of Cys with Ser at position 211 seems to affect the conformational stability of the chitosanase.

• Korean Journal of Organic Agriculture
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• v.14 no.4
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• pp.399-410
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• 2006

This study was carried out to identity the effects of antifungal bacteria isolated from the soil grown kiwi fruit plants on the growth inhibition of Botrytis cinerea causing gray mold in kiwi fruit plants in the southern districts of Jeonnam. Two hundred and fifty antagonistic microorganisms were isolated and examined into the antifungal activity against Botrytis cinerea. We screened and isolated four bacterial strains which strongly inhibited Botrytis cinerea from the soil grown kiwi fruit plants. And the best antifungal bacterial strain which called CHO 163 was finally selected. Antagonistic microorganism CHO 163 was identified to be the genus Bacillus sp. based on the morphological and biochemical characterization. Bacillus sp. CHO 163 showed 86.9% of antifungal activity against Botrytis cinerea. By the bacterialization of culture broth and heated filtrates of culture broth, Bacillus sp. CHO 163 showed almost all of antagonistic activity against Botrytis cinerea. And we also confirmed that in vitro the treatment of Bacillus sp. CHO 163 cultured by SD+B+P broth efficiently controled the growth of Botrytis cinerea causing gray mold in kiwi fruit plants.

• Journal of the Korean Society of Food Science and Nutrition
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• v.17 no.1
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• pp.1-5
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• 1988

Kochujang (Red pepper paste) is one of a traditional fermented seasoning foods having peculia. flavor. In present paper, the effects of microorganisms on the formation of volatile components of Kochujang during fermentation were observed. The volatiles of Kochujang were extracted by a steam-distillation under the reduced pressure and determined by gas chromatography and gas chromatography-mass spectrometry (CC-MS), respectively. During fermentation of Kochujang, Saccharomyces sp. and three strains of Bacillus sp. were isolated as main microorganisms participating in the formation of volatile components. According to the analytical results, tetrahydro geroniol, furfuryl alcohol, methoxy acecoptenone and myrtanal in Kochujang inoculated with Saccharomyces spl, 2-methyl-propan-2-ol, furfuryl alcohol an furfuryl-n-butyrate in Kochujang inoculated with Bacillus sp. B-1, 2-methyl-propan-2-ol, furfuryl alcohol and furfuryl-n-butyrate in Kochujang inoculated with Bacillus sp. B-2, and 2-methyl-propan-2-ol, furfuryl alcohol and n-propylbenzene in Kochujang inculated with Bacillus sp. B-3, were identified as major components, respectively.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.178-182
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• 1989

A gene coding for a xylanase of thermophilic alkalophilic Bacillus sp. K-17 was cloned in Escherichia coli C600 with pBR322. Plasmid pAXl13 was isolated from a transformant producing xylanase, and the xylanase gene was located in a 4.3 Kb HindIII fragment. Biotinylated pAXl13 hybridized to a 4.3 Kb HindIII fragment from chromosomal DNA of thermophilic alkalophilic Bacillus sp. K-17. The xylanase activity was observed in the extracellular curture fluid of E. coli carrying pAXl13. The pAXl13-encoded xylanase had the same enzymatic properties as those of xylanase I produced by thermophilic alkalophilic Bacillus sp. K-17.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.475-478
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• 2003

Bacillus subtilis K-20 chunggukjang is widely used in making soy sauces and bean pastes which are Korean traditional fermented foods. Bacillus subtilis K-20 chunggukjang was cultured, and fermented at $40^{\circ}C$ and 90% humidity for 96 hr after homogenizing with garlic, garlic and onion, and garlic, onion, and ginger. As a result, a product with pizza flavor and taste was obtained from Bacillus subtilis K-20. This product could be used as a functional food to promote immunity.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.603-612
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• 2004

This study was carried out to investigate the replacing fish meal or soybean meal effect of with feather meal or Bacillus sp. inoculated feather meal in broiler diets on the performances and nutrient utilization. One hundred and eighty broilers were randomly allotted to six dietary treatments(1) control, basal diet; (2) BFMl00, Fish meal replacing at 100% level with bacillus sp. inoculated feather meal; (3) BSM20, Soybean meal replacing at 20% level with bacillus sp. inoculated feather meal; (4) BFMl00+BSM20, Fish meal and soybean meal replacing at 100% level and 20% level with bacillus sp. inoculated feather meal, respectively; (5) GFMl00, Fish meal replacing at 100% level with general feather meal; and (6) GSM20, Soybean meal replacing at 20% level with general feather meal) in a 5 week feeding trial. In overall period, body weight gain of control was the highest(1,623g) and those of BFM 100, BFM 100+ BSM 20 and GFM 100 were 1,572g, 1,564g, and 1,078g, respectively. And that of GFM 100(1,078.3g) was the lowest(p<0.05) among treatments. Digestibility of dry matter for BFM 100+BSM 20(81.46%) was higher(p<0.05) than that for other treatments, and that of crude protein was the lowest (p<0.05). Digestibilities of organic matter were significantly(p<0.05) improved when Bacillus sp. inoculated feather meal was replaced at 100% level with fish meal in the basal diet. Methane and hydrogen sulfide gases from the feces were significantly(p<0.05) decreased in chicks fed the control, BFM 100 and GFM 100 diets, when observed after 3 weeks of feeding trials. Feed costs of the control and BFM 100 were 417 and 384 won, respectively but that of BSM 20 was 558 won. Therefore, replacement of fish meal with Bacillus sp. inoculated feather meal in the diets for chicks could be useful for economic production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.5
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• pp.757-763
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• 2010

The qualities of Deodeok Doenjang including physicochemical characteristics and sensory properties were investigated over the following range of Deodeok levels: 5, 10, 15 and 20% (all w/w). The strains used in the manufacturing of Doenjang were Bacillus sp. B-3 and Aspergillus sp. M-9 with the highest enzyme activities like amylase and protease. In case of Deodeok Doenjangs prepared with Bacillus sp. B-3, the amino-type nitrogen of Doenjang containing 10% (w/w) Deodeok was produced more than others during fermentation for 40 days. Amino-type nitrogen produced in Deodeok Doenjang prepared with Aspergillus sp. M-9 was more than one of Deodeok Doenjangs with Bacillus sp. B-3 and a commercial Doenjang. When Deodeok content exceeded 15% (w/w), higher content resulted in lesser amino-type nitrogen production. The results showed that Deodeok had influenced growth of Bacillus sp. and Aspergillus sp. Sensory evaluation showed that Deodeok Doenjangs containing 10% (w/w) Deodeok and 1% (w/w) Bacillus sp. B-3 and containing 20% (w/w) Deodeok and 1% (w/w) Aspergillus sp. M-9 were superior to the other Doenjangs tested.

• Journal of Life Science
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• v.27 no.6
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• pp.688-693
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• 2017

Antifungal bacterium against Colletotrichum coccodes causing black dot disease of potatoes and anthracnose of tomatoes was isolated from sewage sludge. The isolate showed a 99% sequence homology of partial 16S rRNA of Bacillus methylotrophicus CBMB205 and Bacillus amyloliquefaciens subsp. plantarum FZB42. The isolate was identified as Bacillus sp. SW29-2, using the neighbor-joining phylogenetic tree, BlastN sequence analysis, and morphological and cultural characteristics. Bacillus sp. SW29-2 is an aerobic, Gram-positive, endospore-forming bacterium, of which the morphological and physiological characteristics were the same as those of type strain B. lichniformis CBMB205, except for the cell growth of over 4% NaCl. The cell growth of the temperature and the initial pH of the medium was shown at $18-47^{\circ}C$ (opt. ca. $38^{\circ}C$) and 3-9 (opt. ca. 6.0), respectively. The inhibition size (diameter) of Bacillus sp. SW29-2 against four strains of C. coccodes ranged from 23 to 29 mm. Also, the isolate showed antifungal activity against penicillium rot-causing Penicillium expansum in apples. Thus far, any report on the antifungal activity of Baciilus spp. against C. coccodes has not been found. These results suggest that the Bacillus sp. SW29-2 isolate could be used as a possible biocontrol agent against C. coccodes, and further applied to other plant pathogenic fungi.

• Journal of Life Science
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• v.22 no.5
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• pp.605-612
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• 2012

Spore-forming bacteria are being used as probiotic supplements for human and animal use, due to their low pH stability and ability to survive the gastric barrier. In this study, the BCNU 9028 strain was screened from meju, a Korean fermented soybean food starter. Biochemical and physiological characteristics, as well as 16S rDNA sequence analyses, indicate that this strain belongs to the genus $Bacillus$. $Bacillus$ sp. BCNU 9028 showed a 92% survivability at pH 2.5 and could also withstand 0.3% ox bile. Furthermore, it was postulated that $Bacillus$ sp. BCNU 9028 could prevent biofilm formation and adherence of food-borne pathogens such as $Listeria$ $monocytogenes$, $S.$ $aureus$ and $E.$ $coli$ on the basis of its autoaggregation and coaggregation capacity with food-borne pathogens. It was shown that BCNU 9028 has good abilities to adhere to the intestinal tract from its hydrophobic character (63.3%). The $Bacillus$ sp. BCNU 9028 strain especially elicited antibacterial activity against both Gram-positive and -negative pathogens. These findings suggested that the $Bacillus$ sp. BCNU 9028 strain could be used as a potential probiotic.

• The Korean Journal of Food And Nutrition
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• v.26 no.2
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• pp.273-279
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• 2013

The microorganisms producing high protease activity and acid producing ability were isolated from Chunggukjang and kimchi, which were identified as Bacillus subtilis and Lactobacillus planetarum by morphological, biochemical and nutrient requirement. The attempt was made to produce soybean milk yoghurt by using the isolated microorganisms. The mixed culture of Bacillus subtilis and Lactobacillus plantarum exhibited the lowest pH value of 4.23 and highest titratable acidity of 0.88% compared to those of single cultures at $37^{\circ}C$ for 32 hrs, and their total viable count was $4.09{\times}10^8$ $cfu/m{\ell}$. The ${\alpha}$-amylase activity was the highest in culture of Bacillus subtilis after incubation for 24 hrs, while protease activity was most produced in mixed culture of Bacillus subtilis and Lactobacillus plantarum. The amounts of reducing sugars were steadily decreased as soy milk fermentation progressed.