• The Korean Journal of Food And Nutrition
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• v.16 no.4
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• pp.437-443
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• 2003

Chitooligosaccharides were prepared by enzymatic hydrolyzing of crab shell chitosan. Low molecular weight chitooligosaccharides(LMW-chitooligosaccharides), 64.3% of which was composed of trimer, tetramer, and pentamer, was obtained by hydrolyzing chitosan with the chitosanase originated Bacillus pumilus BN-262. High molecular weight chitooligosaccharides(HMW-chitooligosaccharides), 49.3% of which was composed of chitooligosaccharides over heptamer, was obtained by hydrolyzing chitosan with the cellulase originated Trichoderma viride. Acute oral toxicity of chitooligosaccharides were tested in mice. Chitooligosaccharides did not have any toxic effect in mice and oral LD$\_$50/ value of chitooligosaccharides was over 5.0g/kg in mice.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2002.05a
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• pp.165-165
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• 2002

• Microbiology and Biotechnology Letters
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• v.10 no.4
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• pp.245-252
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• 1982

The enzymes lytic to red yeast cell wall, which were produced by Penicillium lilacinum ATCC 36010 and Bacillus pumilus No 41, hydrolyzed an extracellular mannan from Rhodotonla glutinis IFO 0695. mannan was arranged with $\beta$-1,3 and $\beta$-1,4 linkages alternatively. Using this mannan, substrate specificities of these enzymes were investigated. The one from Penicillium lilacinum was an unique mannanase which hydrolyzed $\beta$-1,3 mannoside bond and the other from B. pumilus was a new type of mannanase which cleaved $\beta$-1,4 mannoside bond with requirement of the existence of $\beta$-1,3 linkage on the reducing side. Both enzymes released two kinds of oligosaccharide from mannan, respectively. However, the enzyme from Pen lilacinum produced tetrasaccharide and disaccharide and one of them, tetrasaccharide, was hydrolyzed to disaccharide further. The one from B. pumilus released tetrasaccharide and hexasaccharide from mannan finally.

• Journal of the Korea Organic Resources Recycling Association
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• v.14 no.4
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• pp.141-150
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• 2006

To develop the microbial agents for the environmentally-friendly treatment and recycling of food waste, useful microorganisms, which showed higher degradation activities to various organic compounds and possessed flocculating activities, were isolated from the earthworm. One of the isolated strains, named JS-01, was further selected due to its higher flocculating activity against 0.5% Kaolin clay. JS-01 was identified as Bacillus pumilus sp. by 16s rDNA analysis. The optimal temperature and pH for the growth of JS-01 to express the flocculating activity was found to be at $37^{\circ}C$ in pH 7.0 of EPS broth medium. JS-01 also expressed good degrading activity against cellulose, which is one of the representative organic materials in food waste. We propose that JS-01 will be a good candidate for the efficient treatment of food waste and leachate due to the property to degrade cellulose and flocculating activity.

• 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.

• Korean Journal of Food Science and Technology
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• v.33 no.1
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• pp.84-88
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• 2001

About 100 bacterial strains producing proteolytic enzymes were isolated from Korean traditional soy paste Doenjang. Among them, strain SYG3 producing the highest level of angiotensin converting enzyme (ACE) inhibitor into the culture medium was selected and identified as Bacillus pumilus according to the Bergey's mannual of systematic bacteriology. Soybean powder as a nitrogen source and glucose as a carbon source supported high level of ACE inhibitor production. The presence of 3% NaCl also enhanced the production of ACE inhibitor in the medium. The optimum initial pH of the medium and culture temperature for the production of ACE inhibitor were 7.0 and $32^{\circ}C$, respectively. The maximal level of ACE inhibitory effect was obtained after 36 hours of cultivation under the optimized conditions, which was about 98% of inhibition ratio.

• 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.23 no.11
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• pp.1519-1528
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• 2013

Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.

• 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.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.37 no.3
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• pp.9-16
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• 2005

This study was carried out to investigate the characteristics of extracellular cellulase and xylanase from 4 selected different species, such as enzyme activity and stability by pH, temperature and metal ions, for application into enzymatic deinking system. The optimal temperature and pH for enzyme activity of Bacillus pumilus I, B. subtilis I, B. pumilus II and B. subtilis II were mainly $40{\sim}60^{\circ}C$ and pH $6.0{\sim}7.0$, respectively. Certain metal ions, calcium and cobalt, elevated enzyme activity, even though there were different results of enzyme activities based on various metal ions in 4 different species. With these results we suggest that enzymatic deinking system should be proceed at $50^{\circ}C$ with neutral pH condition.