• The Korean Journal of Food And Nutrition
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• v.12 no.3
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• pp.240-245
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• 1999

The purpose of this study was to develop the new microbial bioflocculant available in a food and fer-mentation industal. This study was reported the results of the composition for optimum culture medium and elemental characteristic of crude purification bioflocculant following the previous report(I). The maximum production of the flocculant from Bacillus megaterium was observated in the culture medium containing 2% sucrose 0.3% NaNO3 0.01% tryptone 0.01% beef extract 0.05% MgSO4 ·7 H2O 0.005% CaCO3 Addition of the sucrose as carbon sources and inorganic salt such as MgSO4, CaCO3 significantly increased the production of flocculant more than nitrogen sources. In the result of color reaction of the crude purified bioflocculant it was investgated that anthrone was positive and benedict burette and nin-hydrin was negative. These result were indicated that the flocculant produced from Bacillus megaterium was a kind of exopolysaccharide.

• Korean Journal of Microbiology
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• v.40 no.1
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• pp.43-48
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• 2004

Bacillus megaterium F7-1 producing keratinolytic protease was isolated from decayed chicken feather. The optimal culture conditions for the production of keratinolytic protease by B. megaterium F7-1 were investigated. The composition of optimal medium for the keratinolytic protease was 0.2% glucose, 0.8% skim milk, 0.05% NaCl, 0.01 % $(K_2HPO_4$, 0.02%, $(KH_2PO_4$ and 0.01 % $MgCl_2$. Especially, skim milk was found to be the most effective compound in keratinolytic protease production. The optimal temperature and initial pH were 6.5 and $25^{\circ}C$, respectively. The keratinolytic protease production under optimal condition reached a maximum of 269 U/ml after 5 days of cultivation. B. megaterium F7-1 degraded 98% of the feather used in the optimized medium within 6 days.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.253-258
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• 2001

A Bacillus circulans KCTC3004 $\beta$-1,3-glucanase gene contained in a recombinant plasmid pLM460 derived from subcloning the original recombinant plasmid pLM530 was trasferred into a new shuttle vector plasmid pLMS1180 by ligating linearized DNAs of pLM460 and pUB110. B. subtilis RM125 and B. megaterium ATCC14945 transformed with pLMS1180 produced the $\beta$-1,3-glucanase substantially. Most of the enzyme was produced during the exponential growth period. The maxium activities of the $\beta$-1,3-glucanase produced by the Bacillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125 (pLM1180) enzyme showed the activity 14 times higher than that of the gene donor cells, followed by the B. megaterium ATCC14945 (pLMS 1180) enzyme with activity 5 times higher than that of the gene donor cells. While E. coli secreted about 7% of the produced enzyme, B. subtilis excreted the enzyme into the medium wholly and B. megaterium about 97% of the total product. The SDS-PAGE of this enzyme produced in E. coli (pLMS1180), B subtilis (pLMS1180) or B. megaterium (pLMS1180) indicated a molecular weight of 38,000. The enzymes overproduced in three different host cells hydrolyzed laminarin to produce mainly laminaribiose, laminaritriose, and laminarioligosaccharides. The plasmid pLMS1180 was stable in B. megaterium, E. coli, but was unstable in B. subtilis.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• The Korean Journal of Food And Nutrition
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• v.11 no.6
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• pp.622-628
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• 1998

Microorganisms isolated from soil were tested for their flocculating activity in kaolin suspension, Identification of the best producing CH-23 strain showed that the strain belonged to the Bacillus megaterium. The maximum production of the flocculating from Bacillus megaterium CH-23 was observed in the culture medium containing 2% sucrose, 3% NaNo3, 0.1% K2HPO4, 0.5% NaCl, 0.5% MgSO4.7H2O and 0.01% tryptone at initial pH 7.0 and 25~3$0^{\circ}C$. Flocculating activity was improved to 57% when the culture medium contained Mn2+(0.01% MnSo4). In the culture medium containing Mg2+(0.01% MgSO4.7H2O) and Ca2+(0.01% CaCO3), flocculating activity were reached to 48% and 33%, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.3
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• pp.403-409
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• 2001

A novel strain of SFO41 producing a large amount of cholesterol oxidase as an extracellular enzyme isolate from Korean salt fermented foods. The strain was identified as Bacillus megaterium based on morphological, cultural and physiological characteristics. Experiments were carried out to optimized the condition of cholesterol oxidase production using B. megaterium SFO41. B. megaterium SFO41 was shown to give the maximum yield of cholesterol oxidase in the medium containing 2.0% glucose, 0.5% yeast extract. 0.03% $MgSO_4{\cdot}7H_2O,\;0.02%\;K_2HPO_4,\;0.2%\;NH_4NO_3$ and 0.2% cholesterol. The optimum culture conditions, temperature, initial pH and agitation speed were $30^{\circ}C$, 7.0 and 150 rpm, respectively. The enzyme production reached a maximum level at 24 hr of cultivation (2.37 U).

• Microbiology and Biotechnology Letters
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• v.27 no.1
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• pp.80-85
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• 1999

To develop bioflocculant for wastewater treatment, about 60 type culture strains and 450 strains isolated from natural sources were examined for screening their ability to flocculate the swine wastewater. Among them, YWO-5 showed the highest activity for NTU removal efficiency and was identified as Bacillus megaterium according to the cultural, morphological and physiological properties. The maximum production of the flocculant was achieved in culture medium containing 2% glucose, 0.05% soytone, 0.01% $CaCl_2$, 0.05% $KH_2PO_4$, and 0.05% yeast extract with initial pH 6.5 when cultured with rotary shaker controlled at 20$^{\circ}C$ and 150 rpm. With jar fermentor, the maximum production was reached to NTU removal efficiency of 93% after 3 days under the optimal conditions. The bioflocculant produced by Bacillus megaterium YWO-5 was effective on various suspended solids and organic wastewaters.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.315-318
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• 2005

The effects of inorganic salts and feather concentrations on pretense production by Bacillus megaterium F7-1 were investigated. Pretense production was dependent on the presence of phosphates in the medium. Supplementation of medium with calcium ion slightly increased protease production. The highest protease production was obtained at $1.4\%$ feather. The optimal medium contained $2.0\%$ glucose, $0.8\%$ skim milk, $0.06\%\;K_{2}HPO_{4}\%,\;0.04\%\;KH_{2}PO{4},\;0.06\%\;NaCl,\;0.03\%\;MgCl_{2}\cdot6H_{2}O,\;0.002\%\;CaCl_{2}\cdot2H_{2}O,\;and\;1.4\%$ whole feather. By using this optimized medium, increased production of the protease was achieved compared with the cases of using basal medium.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.421-424
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• 1992

Escherichia coli JM83 harboring penicilin G acylase gene of Bacillus megaterium ATCC14945 produced a protein in large amount (>20% of the total protein). The protein was identified as GroEL, one of the E. coli heat shock protein, by N-terminal amino acid sequence analysis. It was found that GroEL was induced by the expressed foreign penicilin G acylase at both 27 and $37^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.235-241
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• 2003

For the biological control of Phytophthora blight of red-pepper caused by Phytophthora capsici, an antibiotic-producing plant growth promoting rhizobacteria (PGPR) Bacillus sp. KL 39 was selected from a local soil of Kyongbuk, Korea. The strain KL 39 was identified as Bacillus megaterium by various cultural, biochemical test and API and Microlog system. B. megaterium KL 39 could produce the highest antifungal antibiotic after 40 h of incubation under the optimal medium which was 0.4% fructose, 0.3% yeast extract, and 5 mM KCl at 30 C with initial pH 8.0. The antifungal antibiotic KL 39 was purified by Diaion HP-20 column, silica gel column, Sephadex LH-20 column, and HPLC. Its RF value was confirmed 0.32 by thin-layer chromatography with Ethanol:Ammonia:Water = 8:1:1. The crude antibiotic KL39 was active against a broad range of plant pathogenic fungi, Rhizoctonia solani, Pyricularia oryzae, Monilinia fructicola, Botrytis cinenea, Alteranria kikuchiana, Fusarium oxysporum and Fusarium solani. The purified antifungal antibiotic KL39 had a powerful biocontrol activity against red-pepper phytophthora blight disease with in vivo pot test as well as the strain B. megaterium KL 39.