• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.460-467
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• 2013

This study was conducted to reduce the using amount of chemical fungicides for the control of red-pepper Phytophthora blight. Effect of combination application of two microbial fungicides and two chemical fungicides for the control of red-pepper Phytophthora blight was examined in vitro, in greenhouse and under field conditions. Each microbial fungicides and chemical fungicides was two-fold diluted and mixed-soil drenched. In the greenhouse pot assay, the mixed application of B. pumilus QST2808 and a mixture of dimethomorph + ethaboxam (De) among four mixed applications of two microbial fungicides (B. pumilus QST2808, P. polymyxa AC-1) and two chemical fungicides showed the highest control effect against Phytophthora blight. Also, control effect of mixed application of B. pumilus QST2808 and De was similar to that of single application of De (dimethomorph + ethaboxam) or Mo (mancozeb + oxadixyl). In the field test, when the microbial fungicides (B. pumilus QST2808, P. polymyxa AC-1) and the chemical fungicide(De) for the control of Phytophthora blight of red pepper were mixed-soil drenched four times at 7~10 day-intervals, the control values were in the range of 78.8% to 82.0%. On the other hand when each of the two chemical fungicides (De, Mo) were soil drenched four times at 7~10 day-intervals, the control value were 65.7% to 85.8%. Consequently, the mixed application of the microbial fungicides and chemical fungicides could be recommended as a control method for reducing the using amount of chemical fungicides.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.304-309
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• 2007

Soybean is useful source of protein, especially in Asia. But soybean needs heat inactivation or fermentation process before consumption, since it contains the toxic lectin and various protease inhibitors. Therefore, production of soybean boiling-waste liquor (SBWL) as a byproduct is inevitable. In this study, the chemical composition of SBWL and the optimization of culture conditions for Bacillus pumilus JB-1, a selected strain for functional chungkuk-jang fermentation, using SBWL were investigated. The SBWL contains 88% water, 9.5% free sugar, 1.6% crude protein, 0.3% crude fat, 0.1% crude fiber and 2.1% ash, respectively. The contents of total polyphenol, total flavonoids and free amino acid in SBWL were 55%, 76%, and 30% of those of raw soybean, respectively. Culture conditions for B. pumilus JB-1 in SBWL were optimized. The 1/10-diluted, 0.1 % of $(NH_4)_2SO_4$ added SBWL without pH adjustment and carbon source addition was cultured at $37^{\circ}C$ for 48 h with agitation (120 rpm). The 0.5% of inoculation was enough. The large scale fermentation in 5-L jar fermentor showed that the SBWL is a good resource for production of chungkuk-jang starter and functional ingredients.

• Journal of Life Science
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• v.12 no.2
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• pp.188-199
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• 2002

A gene coding for xylanase from thermo-tolerant Bacillus pumilus TX703 was cloned into Escherichia coli DH5 $\alpha$ using pUC19. Among 7,400 transformants, four transformants showed clear zones on the detection agar plates containing oat-spells xylan. One of them which showed highest xylanase activity was selected and its recombinant plasmid, named pXES106, was found to carry 2.24 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynK) was determined, xynK gene was found to consist of 1,227 base-pair open reading frame coding for a polypeptide of 409 amino acids with a deduced molecular weight of 48 kDa. The coding sequence was preceded by a putative ribosome binding site, the transcription initiation signals, and cia-acting catabolite responsive element. The deduced amino acids sequence of xylanase is similar to those of the xylanases from Hordeum vulgare (barley) and Clostridium thermocellum, with 39 and 31% identical residues, respectively. The amino acids sequence of this xylanase was quite different from those of the xylanases from other Bacillus species.

• Journal of Food Hygiene and Safety
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• v.21 no.3
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• pp.159-163
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• 2006

37 Bacillus spp strains(43.0%=37/86), and 18 B. cereus(20.9%=18/86) were isolated on 86 dried marine products. Each isolates were B. cereus 48.6%(18/37), B. mycoides 13.5%(5/37) B. coagulus 5.4%(2/37) B. firmus 5.4%(2/37), B. circulus 2.7%(1/37), B. stearothermophilus 2.7%(1/37), B. pumilus 2.7%(1/37) B. spp. 8.1%(3/37), and Brebacillus brevis 10.8%(4/37) etc.

• Preventive Nutrition and Food Science
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• v.19 no.3
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• pp.213-219
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• 2014

Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography. The partially purified enzyme was stable between pH 5 and pH 9 and temperature of less than $60^{\circ}C$. Fibrinolytic activity was increased by 5 mM $MgCl_2$ and 5 mM $CaCl_2$ but inhibited by 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM sodium dodecyl sulfate (SDS), and 1 mM ethylenediaminetetraacetic acid (EDTA). The partially purified enzyme quickly degraded the ${\alpha}$ and ${\beta}$ chains of fibrinogen but was unable to degrade the ${\gamma}$ chain.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.558-563
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• 2001

A new alkalophilic strain of Bacillus pumilus JB¬05 producing bleach-stable and halo-tolerant alkaline protease was isolated from cement industry effluents in Karnataka, India. The effects of carbon and nitrogen sources on protease production by this alkalophilic strain were observed after a 30-h incubation. A high level of alkaline protease activity was obtained in the presence of starch as the carbon and peptone as the nitrogen sources. The partially purified enzyme showed an optimum temperature and pH activity at $58^{\circ}C$ and 10.5, respectively. The enzyme was completely inhibited by PMSF (95.0%) indicating it as a serine protease. It is bleach-stable as it retained 35% original activity in the presence of 10% (v/v) hydrogen peroxide at $30^{\circ}$C after 2 h and is halo-tolerant as it retained 70% original activity in the presence of 2.5 M sodium chloride at $30^{\circ}C$ after 2 h incubation.

• The Plant Pathology Journal
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• v.15 no.6
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• pp.319-322
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• 1999

In order to obtain bacterial metabolites inducing disease resistance in pepper plant, two hundred bacterial isolates were isolated from the rhizosphere soil of tobacco, cucumber, and pepper plant. Ethyl acetate extract of each bacterial culture was used to screening for induction of resistance against phytophthora blight of pepper plant. Application of ethyl acetate extract of an isolate SH122 culture to pepper plant conferred resistance against phytophthora blight consistently and significantly. According to cellular fatty acid analysis and other characteristics, the SH122 culture were significantly lower than those on control plants treated with ethyl acetate extract of nutrient broth. The B. pumilus SH122 itself of ethyl acetate extract of its culture did not show antifungal activity against phytophthora blight in pepper plants.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.534-538
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• 2001

The xynA gene encoding an alikali-tolerant endo-1,4-${\beta}$-xylanase (XYN) was cloned from the alkalophilic Bacillus pumilus A-30. The nucleotide sequence of a 974-bp DNA fragment containing the xynA was determined. An ORF of 684 nucleotides that encoded a protein of 228 amino aicds was detected. Asparagine-71 of XYN from B. Pumilus A-30 showed to be highly conservative in alkaline xylanases of family G/11, upon comparing the amino acid sequences of 17 family G/11 xylanases. Site-directed mutation of N71D of the xynA gene resulted in a decrease of 12.4% in the specific acitivity and a significant decline in the enzyme activity in the alkaline pH range.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.154-159
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• 2004

Bacillus species were isolated from swine manure to develope the microbial additive suitable for the rapid com-posting. The 3 of 4 isolated strains were identified as Bacillus cereus KD-2, B. pumilus KD-3, and B. licheni-formis KD-4. Bacillus sp. KD-1 was, however, not highly identical with any Bacillus sp. The isolated strains were analyzed their growth rates, enzyme activities, and antibacterial activities. The maximum growth tem-peratures of KD-1, KD-2, KD-3 and KD-4 were $45^{\circ}C$, $50^{\circ}C$, $53^{\circ}C$, and $55^{\circ}C$, respectively. The activities of pro-tease or amylase in mixed culture of 4 strains were similar in the range of $37^{\circ}C$ to $53^{\circ}C$ and activities of lipase in the range of $37^{\circ}C$ to $42^{\circ}C$ were twice higher than those of lipase in the range of $47^{\circ}C$ to $53^{\circ}C$. The antibacterial activity of KD-l, KD-2, or KD-3 against each other was not detected. That of KD-4 against KD-1, KD-2, or KD-3 was, however, detected. The organic compound and C/N ratio of compost fermented by the mixed culture were determined as 61.9% and 22.4%, respectively. The concentration of the ammonia gas was 12.35 mg/l in the liquid compost.

• Journal of Microbiology and Biotechnology
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• v.24 no.4
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• pp.431-439
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• 2014

We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.