• Korean Journal of Food Science and Technology
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• v.40 no.1
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• pp.76-81
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• 2008

Currently, natural food colorants and preservatives are being used for their general health benefits. Monascus koji, the product of certain fungi that grow on rice grains, has been added to many foods for coloring and preservation. In this study, the antimicrobial activities of Monascus koji ethanol extracts were investigated. Six Monascus strains (M. araneosus KFRI 00371, M. kaoliang ATCC 46597, M. pilosus IFO 4520, M. purpureus IFO 4482, M. ruber IFO 32318 and M. sp. ATCC 16437) were selected based on their relative intensity of red pigment. Two Monascus extracts, M. kaoliang ATCC 46597 and M. purpureus IFO 4482, displayed antimicrobial activities against Bacillus subtilis, B. cereus, Micrococcus luteus, Staphylococcus aureus and Salmonella typhimurium in concentration-dependent manners. The two extracts showed their strongest antimicrobial activity against S. typhimurium, a cause of food poisoning. Therefore, these results suggest that Monascus koji could be used as a natural food colorant and preservative.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.673-679
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• 2004

A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

• Food Science and Preservation
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• v.21 no.3
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• pp.442-450
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• 2014

In this study, we isolated a cellulase-producing bacterium isolated from traditional Korean fermented soybean paste and investigated the effect of culture conditions on the production of cellulase. This bacterium, which was identified as Bacillus subtilis 4-1 through 16S rRNA gene sequence analysis, showed the highest cellulase activity when the cells were grown at $45^{\circ}C$ for 24 hours in the CMC medium supplemented with 1.0% of soluble starch and 0.1% yeast extract. The initial optimum pH of the medium was observed in the range of 5.0~9.0. The optimal pH and temperature for the production of cellulase from B. subtilis 4-1 were pH 9.0 and $60^{\circ}C$ respectively. In addition, the enzyme showed significant activity in the temperature range of $20{\sim}90^{\circ}C$, which indicates that B. subtilis 4-1 cellulase is an alkaline-resistance and thermo-stable enzyme. This enzyme showed higher activity with CMC as the substrate for endo-type cellulase than avicel or pNPG as the exo-type substrates for exo-type cellulase and ${\beta}$-glucosidase. These results suggest that the cellulase produced from B. subtilis 4-1 is a complex enzyme rather than a mono-enzyme.

• Journal of Environmental Science International
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• v.15 no.10
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• pp.983-988
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• 2006

The textile remains have been affected largely by environmental factors including microorganisms because they were composed of organic compounds to be easy to damage. So, we selected 8 strains of the 131 isolated strains from museum environments and textile remains by high pretense activity, and identified them for measuring the antibacterial activity of Gingko biloba extracts. They were identified Genus Arthrobacter spp. 3 strains (Arthrobacter nicotiannae A12, Arthrobacter sp B12, Arthrobacter oxidans B13), Cenus Bacillus spp. 2 strains (Bacillus licheniformis D9, Bacillus cereus D33), Genus Pseudomonas spp. 2 strains (Pseudomonas putida A24, Pseufomonas fluorescene C21) and a Genus Staphylococcus sp. 1 strain (Staphylococcus pasteuri D3) as closest strains through the blast search of NCBI. Though antibacterial activity of the extracts of Gingko biloba leaves as MIC was lower than that of other pharmaceutical antibiotics. However the extracts was crude extracts, the extracts might have good antibacterial against most of the isolates from museum. Especially, the antifungal activity of Gingko biloba is known previously, the extracts of Gingko biloba leaves has possibility of usage as a good natural material for conservation of remains.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.9
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• pp.1237-1242
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• 2009

This study was conducted to isolate strain for the preparation of fermented antler (Cervus cornu parvum) and evaluate its physiological activities. The growth degrees of twenty-one samples from Bacillus sp., Lactobacillius sp. and mushroom strain on antler extract agar were evaluated in this study, and Bacillus subtilis KH-15, SCB-3, Cordyceps militaris, Phellinus linteus, Inonotus obliquus 26136, and Inonotus obliquus 26147 were selected. The fermented antler extract by C. militaris had relatively higher contents of total sugar (1619.3 ${\mu}g$/mL), uronic acid (302.0 ${\mu}g$/mL), sulfated-glycosaminoglycan (S-GAGs) (119.9 ${\mu}g$/mL) and sialic acid (21.6 ${\mu}g$/mL) than any other extracts. The anti-complementary activities of all fermented antler extracts were higher than non-fermented antler extract, and among these samples, fermented antler extract by C. militaris showed the highest anti-complementary activity (inhibition of 50% total complement hemolysis, $ITCH_{50}$; 50.1% at 1,000 ${\mu}g$/mL). The ability of fermented antler extract by B. subtilis KH-15 to scavenge 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical ($IC_{50}$; 4.97 mg/mL) was significantly the highest (p<0.05), whereas the extract from I. obliquus exerted significantly (p<0.05) high 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity ($IC_{50}$; 16.98 mg/mL) among all samples. The results of this study suggest that physiological effects including immuno-modulating and antioxidant activities of the antler may be increased through fermentation process.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.549-554
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• 2006

To standardize a manufacturing method and improve the quality of traditional kochujang, eight-types of meju with different shapes (brick, grain) were prepared using Aspergillus oryzae (A.o) or Aspergillus sojae (A.s) alone or in combination with Bacillus subtilis (B.s). The physicochemical characteristics and enzyme activities of the various meju were compared during fermentation for 12 days at $28^{\circ}C$. The moisture content of both the brick- and grain-shaped meju were gradually decreased from an initial content of 50.47 to 54.89% to a content of 12.91 to 16.25% on day 12 of fermentation. The neutral protease activities of the brick-shaped meju ranged from $1.19{\pm}0.12$ to $1.25{\pm}0.28\;unit/mL$, and were similar for all treatments. The ${\alpha}$-amylase activities in A.s+B.s treatment of brick-shaped and grain-shaped meju were the highest, $11{\pm}0.6$ and $9{\pm}0.7\;unit/mL$, respectively. The ${\beta}$-amylase activities ranged from $1.53{\pm}0.01$ to $1.56{\pm}0.02\;unit/mL$, and were similar for all treatments. The amino type nitrogen content of A.o+B.s brick-shaped meju was the highest, $0.39{\pm}0.03%$. We confirmed that the brick-shaped meju prepared with A. oryzae and B. subtilis could be used to prepare traditional kochujang to improve the quality of the product.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.35-42
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• 2003

A 1,3 kb cellulase gene encoding novel bifunctional cellulase-chitosanase activity was cloned from biopolymer-producing alkali-tolerant B. lichenformis NBL420 in E. coli. A recombinant cellulase-chitosanase, named CelA, was expressed and purified to homogeneity. The activity staining and the enzymatic characterization of the purified CeIA revealed bifunctional activities on carboxymethyl cellulose (CMC) and glycol-chitosan. The similar characteristics of the enzymatic activities at the optimum pH, optimum temperature, and thermostability Indicated that CelA used a common catalytic domain with relaxed substrate specificity. A comparison of the deduced amino acids in the N-terminal region revealed that the mature CelA had a high homology with the previously identified bifunctional cellulase-chitosanase of Myxobacter sp. AL- 1.

• Journal of Life Science
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• v.12 no.6
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• pp.700-707
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• 2002

To study the characteristics of organic and nutrient removal by Bacillus species at high COD concentration of influent, three lab-scale batch reactors(R1, R2, R3), each of which has different substrate composition, were operated. More than 95% of $NH_4^+$-N and $COD_{cr}$, concentrations were removed under an aerobic condition, and their removal efficiencies were found to be 22.6 and 90.5%(R1), 23.9 and 65.8%(R2), 30.2 and 86.4%(R3), respectively. The removal efficiency of $NH_4^+$-N was high when an enough amount of $NO_3^{-}$-N was supplied, and that of $COD_{cr}$. was low when a high concentration of initial $NO_2^{-}$-N was added. The amount of carbon utilized in denitrification was a little. In all reactors,$NO_3^{-}$-N was removed under an anoxic condition, but in the R3 reactor, 10% of $NO_3^{-}$-N could be removed even undo, an aerobic condition. The removal efficiencies of TN and TP were 41.8 and 49.5%(R1), 40.1 and 35.8%(R2), 47.0 and 57.6%(R3), respectively. Alkalinities destructed under an aerobic condition for each reactor were 4.96, 5.41 and 3.93 mg/L (as $CaCO_3$) per each gram of $NH_4^+$-N oxidized, respectively, while 3.06, 3.17 and 2.60 mg/L (as $CaCO_3$) of alkalinities were produced for each gram of ,$NO_3^{-}$-N reduced to $N_2$. The SOUR were found to be 38.5, 52.7 and 42.0 mg $O_2$/g MLSS/hr, which indicated that Bacillus sp. had a higher cell activity than activated sludge. The OLR and sludge production were estimated to be 0.69 and 0.28(Rl), 0.77 and 0.20(R2), 0.61 kg COD/$m^3$/day and 0.25 kg MLSS/kg COD(R3), respectively. From the N-balance, the highest percentage(40.9%) of nitrogen lost to $N_2$ was obtained in the R3 reactor. From all the results, the possibility of aerobic denitrification Bacillus sp. has been shown and the B3 process seemed to have two advantages: a little amount of carbon was required in denitrification and not much amount of alkalinity was destructed under an aerobic condition.

• Korean Journal of Food Science and Technology
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• v.44 no.5
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• pp.600-606
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• 2012

A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.