• Journal of Life Science
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• v.22 no.7
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• pp.912-919
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• 2012

A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.

• Food Science and Biotechnology
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• v.14 no.5
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• pp.704-708
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• 2005

Poly-y-glutamate (${\gamma}$-PGA) has great potential as a biodegradable polymer in a broad range of industrial fields such as food, cosmetics, medicine and water treatment. In order to isolate ${\gamma}$-PGA producers that are suitable for specific industrial applications, 653 Bacillus-like strains were isolated from 439 varieties of three Korean fermented foods, Cheongkukjang, Doenjang, and Kochujang, which were collected from different regions across Korea. A very high level of ${\gamma}$-PGA production was demonstrated in 4.7%, 1.8%, and 3.0% of the Bacillus-like strains isolated from Cheongkukjang, Doenjang, and Kochujang samples, respectively, which produced a viscous substance to such extent that it overflowed to the lid of the plate on the glutamate-dependent ${\gamma}$-PGA production plates. On glutamate-independent ${\gamma}$-PGA production plates, 5.1%, 5.9%, and 6.1% of Bacillus-like strains isolated from Cheongkukjang, Doenjang, and Kochujang samples, respectively, showed high production. The maximum ${\gamma}$-PGA production yields were 32.5 g/L and 5 g/L, depending on the purification methods in the glutamate-dependent media, with the higher yield resulting from a simple precipitation of ${\gamma}$-PGA by either methanol or ethanol and dialysis. The viscous substance produced by each strain showed different morphological characteristics, suggesting that isolated ${\gamma}$-PGA producers could produce various types of ${\gamma}$-PGA.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1078-1085
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• 2018

A salt-tolerant cellulase secreted by a marine Bacillus sp. SR22 strain with wide resistance to temperature and pH was purified and characterized. Its approximate mass was 37 kDa. The endoglucanase, named as Bc22Cel, was purified by ammonium sulfate precipitation, gel filtration chromatography, and extraction from the gel after non-reducing sodium dodecyl sufate-polyacrylamide gel electrophoresis. The optimal pH value and temperature of Bc22Cel were 6.5 and $60^{\circ}C$, respectively. The purified Bc22Cel showed a considerable halophilic property, being able to maintain more than 70% of residual activity even when pre-incubated with 1.5 M NaCl for 1 h. Kinetic analysis of the purified enzyme showed the $K_m$ and $V_{max}$ to be 0.704 mg/ml and $29.85{\mu}mol{\cdot}ml^{-1}{\cdot}min^{-1}$, respectively. Taken together, the present data indicate Bc22Cel as a potential and useful candidate for industrial applications, such as the bioconversion of sugarcane bagasse to its derivatives.

• The Korean Journal of Pesticide Science
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• v.19 no.1
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• pp.71-79
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• 2015

In order to investigating the effects of antifungal activity of intestinal bacteria obtained from insect, it was identified these bacteria isolated from the gut. In this result, total 49 isolates of intestinal bacteria were identified from 10 kinds of insect species. It was that 4 isolates including Cedecea sp. from Nesidiocoris tenuis, 3 isolates including Enterobacter sp. from Odontotaenius disjunctus, 4 isolates including Acinetobacter sp. from Reticulitermes speratus, 4 isolates including Clavibacter sp. from Riptortus clavatus, 11 isolates including Bacillus sp. from Lema decempunctata, 3 isolates including Enterococcus sp. from Henosepilachna vigintioctopunctata 2 isolates including Staphylococccus sp. from Harmonia axyridis, 5 isolates including Enterobacter asburiae from Popillia mutans, 7 isolates including Aeromonas sp. from Hydrophilus acuminatus, and 7 isolates including Brucella sp. from Anomala octiescostata. In order to investigating antifungal activity against plant-pathogenic fungi, Altanaria solani, Colletotrichum gloeosporioides, Botrytis cinerea, Fusarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Selerotinia sclerotiorum were dual cultured with each 49 gut enterobacteriaceae. As these results showed that many isolates have the antifungal activities including 26 isolates against A. solani, 6 isolates against B. cinerea, 13 isolates against C. gloeosporioides, 11 isolates against F. oxysporum, 17 isolates P. capsici, 2 isolates against R. solani and 2 isolates against S. sclerotiorum. Pseudomonas aeruginosa was showed strong antifungal activity against all of tested plant pathogens. It might be taken a potential for application against plant-pathogenic fungi with useful control agent.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.618-624
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• 1998

The culture conditions for Bacillus sp. DP-152 in the flask were investigated for the production of polysaccharide locculant, DP-152. The optimum pH and temperature for the locculant production were 8.0 and $30^{\circ}C$, respectively. The avorable substrates for flocculant production were soluble tarch and ammonium nitrate. The medium composition was optimized as follows: 30 g soluble starch, 0.75 g $NH_4NO_3,\; 2.0g\; K_2\;HPO_4,\; 0.1\; g KH_2PO_4,\; 0.2g\; MgSO_4.\; 7H_2O,\; and\; 0.2g\; MnSO_4~5H_2O$ in 11 of distilled water. Under this optimized condition, flocculating activity has been improved 4-fold compared with that of the basal medium. In the culture flask, the highest flocculating activity was obtained after 70 h of cultivation and the amount of bioflocculant DP-152 yielded was 12.4 g/$\ell$. The solution of bioflocculant DP-152 showed non-Newtonian characteristics. Bioflocculant DP-152 exhibited apparently higher viscosity at all concentrations compared to that of zooglan (from Zoogloea ramigera), and it was stable over a wide range of temperatures and pHs.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.578-584
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• 1990

A microorganism capable of producing high level of extracelluar cyclomaltodextrin glucanotransferase(EC 2.4.1.19; CGTase) was isolated ’rom soil. The isolated strain No. 239 was identified as Bacillusstearothermophilus. The maximal CGTase production (about 7.0 unitslml) was observed in medium containing2% soluble starch, 0.5% defatted soybean meal, 0.1% NaH_2PO_4.2H_2O$ and 0.015% $ CaC_l2 $ with initial pH 7.0. The strain was cultured at $55^{\circ}C$ for 48 hr with reciprocal shaking. At 0.83% substrated concentration potato starch was the optimum substrate with 50.1% conversion to cyciodextrin (CD)after the reaction at $65^{\circ}C$ for 24 hr (CGTase 10 unitlg starch). Using soluble starch as substrate (5% substrate concentration, CGTase 10 unitlg starch), the maximum conversion of 40% was obtained at11 hr reaction, and the ratio of $\alpha-, \beta-$ and $\gamma$-CD production at this time were 1.0:1.3:0.4, respectively., respectively.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.432-439
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• 2021

Microbial community plays important roles in the culture environment of mud crab Scylla serrata. One of the environmental management efforts for the cultivation of S.serrata is by stabilizing microorganisms involved in nitrogen cycle process. The availability of dissolved inorganic nitrogen in its culture environment under a recirculating system closely relates to the nitrogen cycle, which involves both anaerobic and aerobic bacterial activities. Anaerobically, there are two major nitrogen compound degradation processes, i.e., denitrification and dissimilatory nitrate reduction to ammonium (DNRA). This study aimed to identify denitrifying and DNRA bacteria isolated from the recirculating cultivation of S. serrata. The water samples were collected from anaerobic filters called close filter system, which is anaerobically conditioned with the addition of varying physical filter materials in the recirculating mud crab cultures. The results showed that three denitrifying bacterial isolates and seven DNRA bacterial isolates were successfully identified. The phylogenetic analysis based on 16S rRNA gene of the denitrifying bacteria revealed that HIB_7a had the closest similarity to Stenotrophomonas daejeonensis strain MJ03. Meanwhile, DNRA bacterial isolate of HIB_92 showed a 100% similarity to Bacillus sonorensis strain N3, Bacillus vallismortis strain VITS-17, Bacillus tequlensis strain TY5, Geobacillus sp. strain DB24, Bacillus subtilis strain A1, and Bacillus mojavensis strain SSRAI21. This study provides basic information denitrifying and DNRA bacterial isolates identity which might have the potential to be applied as probiotics in aquaculture systems in order to maintain optimal environmental conditions.

• Applied Biological Chemistry
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• v.34 no.4
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• pp.307-311
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• 1991

A moderate halophile, ES 10 which produces a high level of alkaline protease was isolated from the salted anchovies and indentified as a strain of Halomonas sp. The optimum growth of the Halomonas sp. was revealed in the presence of 2 M NaCl and its growth rate in the Temporary Synthetic Medium was increased by adding DL-alanine, but inhibited by adding L-proline. The concentration of $Na^+$, $K^+$ and $Mg^{2+}$ in the cell mass of the Halomonas sp. ES 10 was 5-, 25- and 35-fold higher by dry weight basis, respectively than those of B. subtilis or E. coli. Norberg and Hofsten medium with 1 M NaCl was selected as the best medium for producing high level of alkaline protease. The optimum temperature for the growth and protease production was equally $20^{\circ}C$.

• Journal of Sericultural and Entomological Science
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• v.24 no.1
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• pp.32-38
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• 1982

The study has been carried out to investigate a possibility to control the pine needle gall midge, Thecodiplosis japonensis Uchida et Inoue, by microbial pathogens as one of the microbial control measures. The samples were collected at Kocheon-Ri in the suburbs of Suweon city where were heavily damaged by this pest. Microorganisms were isolated from the both of diseased larvae and baiting method of soil microbes. In addition to, several species of the silkworm mucardine diseases were isolated for their infectivity of these fungi to the larvae of pine needle gall midge. Six species of fungi, Aureobasidium pullulans, Ascochyta sp, Verticillium psalliotae, Streptomyces sp., and two species of Aspergillus were isolated from the galls and soils, five species of muscardine diseases, Isaria farinosa, Spicaria pracina, Oospora destructor, Aspergillus flavus and A. oryzae were also identified from the silkworm corpse collected in the silkworm rearing farmers. Total of eleven species of fungi identified from the both of the larval of pine needle gall midge and silkworm larvae were tested for their pathogenecity to the larvae of pine needle gall midge. All of eleven species of fungi identified showed a considerable infectivity to the larvae. Twenty nine different kinds of bacteria were isolated from the both of diseased larvae and infested soils through the direct planting for the larvae and streaking for the corpse. The infectivity test is in progress. However two kinds of bacteria were recognized to be pathogenic to the larvae tested.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.106-107
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• 2003

The effect of biological control agent Bacillus sp. (BAC03-3-1, BAC03-3-2, BAC02-4) on pre- and postemergence Sclerotinia rot of perilla (Perilla frutescens var. japonica) caused by Sclerotinia sclerotiorum was determined from greenhouse field trials. The ability of this antagonist to reduce germination of sclerotia of S. sclerotiorum was also evaluated. In the greenhouse, suspension of BAC03-3-1 application as root drench of perilla, which provided as little as 10$\^$7/ cells/ $m\ell$ per gram of soil, significantly increased plant stand in pathogen-infested soil over that in the untreated control. All three isolates reduced the germination of sclerotia of S. sclerotiorum in loamy sand soils in the greenhouse. In loamy sand amended with rice bran the sclerotial germination was inversely correlated (r = -0.79) with perilla stand in the greenhouse. However, a higher rate of bacterial suspension with rice bran(Ig dwt./100g soil) than that applied with bacterial suspensions only was necessary to achieve a comparable reduction in sclerotial germination. In field study, all three isolates added to soil to provide 10$\^$7/ cells/$m\ell$ per gram significantly prevented Sclerotinia rot (73-85％) after 35 days of growth. The isolate BAC02-4, BAC03-3-1 and BAC03-3-2 gave final stands of 65 to 75, 60 to 70, and 55 to 60％, respectively. The addition of rice bran(1 ％) to loamy sand in the field resulted in a 10-fold increase in propagule numbers of the three isolates within 10 days of application.