• Journal of Life Science
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• v.16 no.3 s.76
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• pp.420-426
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• 2006

Bacillus sp. KMU-991 was isolated from Oslo city soils at Norway and shown a strong antifungal activity on red-pepper anthracnose, Colletotrichum gloeosporioides. Bacillus sp. KMU-991 produced a maximum level of antifungal substrate under aerobic incubation at $30^{\circ}C$, 180 rpm for 48 hours in TSB medium(initial pH 7.0) containing 1.0% mannitol and 1.0% ammonium chloride. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against C. gloeosporioides KACC 40804. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Fusarium oxysporum f. sp. radicus-lycopersici KACC 40537, Rhizoctonia solani AG-4 KACC 40142, Botrytis cinerea KACC 40573, Colletotrichum orbiculare KACC 40808, and Phytophthora cambivora KACC 40160 by agar diffusion method.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.85-91
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• 2001

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

• Journal of Life Science
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• v.6 no.2
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• pp.79-86
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• 1996

A bacterium producing pullulanase was from soil, and was identified Bacillus cereus and named as Bacillus cereus JK36. The optimal culture conditions for the efficident production of pullulanase from B. cereus JK36 was obtained by cultivating with the medium composed of 1% pullulan, 1% teast extract, 1% bactopeptone, 0.1% NaH$_{2}$PO$_{4}$, 2H$_{2}$O, 0.02% MgSO$_{4}$\ulcorner7H$_{2}$O at 40$\circ$C, initial pH 6.5 for 70 hours. Using the culture supernatant as crude enzyme, the optimal pH and temperature of the pullulanase of this strain were 6.5 and 50$\circ$C. In effect of pH and temperature on the stability of the enzyme, the enzyme was stable in the range of pH6.0$\sim$9.5 and up to 40$\circ$C, respectively. The hydrolysis product on pullulan was mainly maltotriose.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.484-489
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• 1997

The characteristics of growth and esterase activity of bacterial strains isolated from acid hydrolysed soybean protein were examined. All the isolated strains having decomposition activity of p-hydroxybenzoic acid butyl ester and esterase producing activity were identified as Bacillus sp. by morphological and biochemical methods. The specific growth rates, esterase activities and p-hydroxybenzoic acid butyl ester decomposition activities of isolated strains were $0.844{\sim}1.213\;h^{-1}$, $21{\sim}222\;mU/ml$ and $5.4{\sim}8.1\;mU/ml$, respectively. In the fermentation of Bacillus sp. KB8 strain which had the highest esterase producing activity, growth, extracellular excretion and intracellular synthesis of esterase were inhibited by adding NaCl in the culture broth. Esterase producing activity gradually increased after late exponential growth phase, until maximum value of 420 mU/ml reached after 64 hours culture period. Esterase of Bacillus sp. KB8 strain was stable up to $50^{\circ}C$ for 30 minutes, but was inactivated by heating for 30 minutes at $70^{\circ}C$. The enzyme activity exponentially decreased during the incubation time at the temperatures of $60^{\circ}C$ and $65^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.406-411
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• 2008

This study was conducted to examine the quality enhancement of fermented soybean pastes (cheonggukjang) using microorganisms with high enzyme activities and proper experimental design. The microorganisms for soybean paste fermentation were selected from a specific area of Gyeonggi and were idenlified by 16S rDNA sequence analysis. To prepare the cheonggukjang, an optimum mixing ratio of selected microorganisms was determined using contour plots and numerical optimization methods. A total of 39 microorganisms were isolated from the soybean paste, consisting primarily of Bacillus subtilis and Bacillus licheniformis, and no mold was found. Three microorganisms showing high enzyme activities were selected and used to formulate an optimum mixing ratio for cheonggukjang preparation. Based on levels of amino-nitrogen, ammonium-nitrogen, antioxidant activity values, and sensory preference results, the optimum mixing ratio of 50% of Bacillus sp. SC-1 and 50% SC-3 was suggested for the manufacture of high quality of cheonggukjang.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.327-333
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• 2014

Two genes encoding the mannanase of Bacillus sp. YB-1401 and B. amyloliquefaciens YB-1402, which had been isolated at acidic pH as mannanase producers, were each cloned into Escherichia coli, and sequenced. Both mannanase genes consisted of 1,080 nucleotides, encoding polypeptides of 360 amino acid residues. The deduced amino acid sequences of the two mannanase genes differed by four amino acid residues different, and were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. Comparison of two mannanases produced from recombinant E. coli indicated that His-tagged mannanase of YB-1402 (HtMAN1402) was more stable than that of YB-1401 at acidic pH and high temperature. In particular, HtMAN1402 retained more than 50% of its activity at pH 3.0 after 4 h of pre-incubation, suggesting the enzyme is a valuable candidate for use as a feed additive. In addition, thermostability of the two mannanases was found to be enhanced by $Ca^{2+}$ ions.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.8 no.3
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• pp.627-631
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• 2007

The factors affecting on sludge sedimentation are reported as F/M ratio, ingredient, composition of influent substrate, dissolved oxygen concentration, temperature, pH, filamentous bacteria and SRT. Aeration tank applying Bacillus sp. has an important role for maintaining the dominant microorganism species to make steady progress for spore growth affecting sedimentation. This research aims to investigate the affecting factor for the sedimentation in B3 system and RABC system with aeration tank applying tapered aeration. Extracellular polymeric substances(EPS), protein and carbohydrate can be produced for the extreme condition, that is down to 0.2 mg/L of dissolved oxygen in the aeration tank. This research found out the relation between the sedimentation and the EPS production, especially the ratio of protein/carbohydrate. The spore of Bacillus sp. was formed at the low DO then microorganisms produced EPS. The results showed that the production of EPS was 109.95 mgEPS/gSS at 1.6 mg/L of DO, however it was 131.77 mgEPS/gSS at 0.5 mg/L of DO. The sedimentation was affected by protein content in EPS and the ratio of protein and carbohydrate. The settleability of sludge was not affected by the ratio of protein/carbohydrate in B3 process, meanwhile settleability was affected by the ratio of it in RABC process, respectively.

• Journal of Life Science
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• v.17 no.11
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• pp.1601-1604
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• 2007

The gene for ${\beta}-agarase$ of an Agarivorans sp. JA-1 was expressed in Bacillus subtilis DB104, 168 and ISW1214 strains for mass-production. Among 3 host strains, B. subtilis ISW1214 secreted the highest amount of recombinant ${\beta}-agarase$ with a specific activity of 201 U/mg and 360 mg of protein into culture broth. This was approximately 130-fold higher than the production in E. coli as an expression host. Recombinant enzyme produced neoagarooligosaccharides such as neoagarohexaose, neoagarotetraose, and neoagarobiose from agar. Produced neoagarooligosaccharides showed antibacterial activities against gram-negative E. coli and gram-positive B. subtilis at a concentration of 1.5%. These data suggest that neoagarooligosaccharides could be an useful preservative for food industry.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.39-48
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• 2017

In order to evaluate the diversity of microbial population of Korean traditional Deonjang and Gochujang produced in Jeju, Jeonnam, and Jeonbuk province area, microbial communities were analyzed using next generation sequencing. In this result, the dominant bacteria of Deonjang in three area were Bacillus amyloliquefaciens, Tetragenococcus halophilus, and Bacillus was major dominant bacteria in Jeonnam (43.16%) and Jeonbuk (64.54%) area. But in Jeju area, Bacillus was 0.22%, which was significantly different from the other two. Equally, the dominant fungi of Deonjang in 3 area were Candida versatilis. Common fungus in Jeonnam and Jeonbuk area was Candida sp., respectively, 64.22% and 33.68% and Micor sp. was a common fungus in Jeonnam (15.66%) and Jeonbuk area (36.73%). But in Jeju area, Candida sp. and Zygosaccharomyces rouxii were dominant than mold. Bacillus subtilis, Bacillus licheniformis, and B. amyloliquenfaciens were the preminant bacteria in the traditional Gochujang in three regions. But there were no common dominant fungi in the 3 regions. Aspergillus sp. and Rhizopus sp. prevailed in Jeju and Jeonnam region, and Zygosaccharomycess rouxii predominanted in Jeonbuk area. These results suggested that the difference in the samples collected for the study were classified into similar groups according to the characteristics of each sample rather than regional characteristics.

• Journal of Microbiology and Biotechnology
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• v.24 no.8
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• pp.1073-1080
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• 2014

A total of 10 cellulase-producing bacteria were isolated from soil samples irrigated with paper and pulp mill effluents. The sequencing of 16S rRNA gene revealed that all isolates belonged to different species of genus Bacillus. Among the different isolates, B. subtilis IARI-SP-1 exhibited a high degree of ${\beta}$-1,4-endoglucanase (2.5 IU/ml), ${\beta}$-1,4-exoglucanase (0.8 IU/ml), and ${\beta}$-glucosidase (0.084 IU/ml) activity, followed by B. amyloliquefaciens IARI-SP-2. CMC was found to be the best carbon source for production of endo/exoglucanase and ${\beta}$-glucosidase. The ${\beta}$-1,4-endoglucanase gene was amplified from all isolates and their deduced amino acid sequences belonged to glycosyl hydrolase family 5. Among the domains of different isolates, the catalytic domains exhibited the highest homology of 93.7%, whereas the regions of signal, leader, linker, and carbohydrate-binding domain indicated low homology (73-74%). These variations in sequence homology are significant and could contribute to the structure and function of the enzyme.