• KSBB Journal
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• v.21 no.1 s.96
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• pp.42-48
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• 2006

Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

• Food Science and Biotechnology
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• v.15 no.2
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• pp.214-219
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• 2006

A bacterial strain, initially identified as B1-3, was isolated from cheonggukjang, a traditional Korean dish made from fermented soybeans. Using the Biolog system and 16S rRNA sequence analysis, we identified B1-3 as Bacillus mojavensis. We manufactured a quickly fermented soybean (QFS) food product using the B. mojavensis, and guided by their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging ability. We isolated substances with antioxidative activity from it. Using mass spectrometry (MS) and nuclear magnetic resonance (NMR) techniques, we isolated 4 compounds from the ethyl acetate (EtOAc)-soluble neutral fraction of methyl alcohol (MeOH) extracts of the QFS food product (genistein, daidzein, 3R,4R-3-methyl-3,4-dihydroxy-2-pentanone, and 3S,4R-3-methyl-3,4-dihydroxy-2-pentanone) and 3 compounds from its acidic fraction (4-hydroxyphenylacetic acid, genistin, and daidzein). Two compounds from the neutral fraction (3R,4R-3-methyl-3,4-dihydroxy-2-pentanone and 3S,4R-3-methyl-3,4-dihydroxy-2-pentanone) were not detected in nonfermented soybeans (NFS) or in the filtrate of the LB broth used to culture B. mojavensis. However, they were detected in the filtrate of the same broth when it contained 2% glucose. These results suggest that these 2 compounds were derived from glucose (or other saccharides) in the soybean during fermentation. One compound that was found in the acidic fraction (4-hydroxyphenylacetic acid) was readily detected in NFS, but not in the culture broth. This suggests that 4-hydroxyphenylacetic acid was derived from NFS. We concluded that the antioxidative activity of cheonggukjang is a result of the interactions between soybean components and the microorganisms used in the fermentation of cheonggukjang.

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.32-32
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• 2023

Cadmium and salt exposure to crops is considered vulnerable for production as well as consumption. To address these challenges, the current study aimed to mitigate the toxicity induced by salt and cadmium in soybean plants through the application of bacterial strain Bacillus safensis KJW143 isolated from the rhizosphere of oriental melon..The bioassay analysis revealed that KJW143 is a highly salt-tolerant and cadmium-resistant (Cd) strain with an innate ability to produce melatonin, gibberellin (GA3), Indole-3-Acetic Acid (IAA), and organic acids (i.e., acetic, succinic, lactic, and propionic acids). Soybean plants at 20 days old were treated with KJW143 in a different form (pellet, broth, and together) and their effect on plant performance was investigated. Inoculation with KJW143enhanced plant biomass and growth attributes in soybean plants compared to the control (non-treated). In particular, we observed that only pellet-treated showed 65%, 27.5%, and 28.7% increase in growth (shoot fresh weight) compared to broth, broth with pellet, and control. In addition, bacterial strain KJW143 treatment (only pellet) modulated the physiochemical apparatus of soybean plants by increasing glucose (390%), arabinose (166%), citric acid (22.98%) and reducing hydrogen peroxide (29.7%), catalase (32.1%), salicylic acid (25.6%) compared to plants with combined stressed plants (cd and salinity). These findings suggest that bacterial strain KJW143 could be usedas a biofertilizer to minimize the probable risk of heavy metal and salinity stress on crops.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.420-426
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• 2006

Bacillus sp. KMU-991 was isolated from Oslo city soils at Norway and shown a strong antifungal activity on red-pepper anthracnose, Colletotrichum gloeosporioides. Bacillus sp. KMU-991 produced a maximum level of antifungal substrate under aerobic incubation at $30^{\circ}C$, 180 rpm for 48 hours in TSB medium(initial pH 7.0) containing 1.0% mannitol and 1.0% ammonium chloride. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against C. gloeosporioides KACC 40804. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Fusarium oxysporum f. sp. radicus-lycopersici KACC 40537, Rhizoctonia solani AG-4 KACC 40142, Botrytis cinerea KACC 40573, Colletotrichum orbiculare KACC 40808, and Phytophthora cambivora KACC 40160 by agar diffusion method.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.451-456
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• 2003

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

• Applied Biological Chemistry
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• v.31 no.4
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• pp.356-360
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• 1988

The alkaline protease producing bacteria isolated from soil and identified as Bacillus subtilis. The optimum medium for alkaline protease production from the microorganism was as follows; soluble starch, 1.5% ; proteose peptone, 0.5% ; $K_2HPO_4$, 0.1% ; $MgSO_4{\cdot}7H_2O$, 0.02% and sodium carbonate, 1.0%. The optimum temperature for alkaline protease production was $35^{\circ}C$, and the initial pH of medium was pH 10.5. The alkaline protease activity was about 2,300 U per ml of culture broth by Casein-Folin Method. A 9.2 fold purification of alkaline protease was obtained from culture broth. The recovery was 14% and purified enzyme was identified as single band, and its molecular weight was about 19,000. The optimum temperature for enzyme reaction was $70^{\circ}C$, and optimum pH was 12. The activity of purified enzyme was inhibited by metal ion ($Fe^{++}$), and Phenylmethylsulfonyl Fluoride, a serine protease inhibitor.

• KSBB Journal
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• v.15 no.1
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• pp.35-41
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• 2000

We tried to prepare encapsulated whole cell cyclodextrin glucanotransferase(CGTase) in order to produce glycosyl-xylitol using xylitol as glucosyl acceptor. The organic nitrogen source was more effective for the production of CGTase from Bacillus macerans IFO 3490 than the inorganic one. Most of the CGTase which had been produced during cultivation was excreted to the growth medium. B. macerans cells inocculated in the capsule failed to grow to the high cell density. Adsorbents such as activated charcoal, Sephadex and Amberite resins could not adsorb efficiently the CGTase from the broth solution. We obtained successfully the encapsulated whole cell CGTase by immobilizing the concentrated broth solution in the calcium alginate capsules. The encapsulated whole cell CGTase carried out the transglycosylation reaction which converts xylitol into glucosyl-xylitol using dextrin as glucosyl donor.

• Journal of Naturopathy
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• v.7 no.2
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• pp.63-69
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• 2018

Purpose: The purpose of this study was to investigate the effects of Bacillus fermentation broth (ENM) on the human NK cell activity, and bone matrix density, matrix content and area of hamsters fed ENM. Methods: NK cell activity was tested, and bone mineral density were measured by x-ray. Results: NK cell activity was significantly higher in the control group (644.71 pg/ml) and in the test group (1796.37 pg/ml) (p<.004). Test groups were significantly increased by 1110.37 pg/ml in the pre-test and 1796.37 pg/ml in the post-test(p<.001). Behavioral observations after feeding ENM to hamsters showed normal behavior with no difference between control and test groups. The hamster body weight of the control was 106 g at the initial and final 27 days. In the test group, the initial weight was 96.6 g and the final 27 days was 114 g. No morphological changes were observed in the X-ray photographs of the hamster hind legs. The bone matrix density was 0.059 g/cm³ in control and 0.062 g/cm³ in the test, which was increased by 0.003 g/cm³. The bone matrix content was 0.175 g in the control and 0.196 g in the test. The bone area was 2.95 cm² in the control and 3.19 cm² in the test, which was increased by 0.19 cm². In the autopsy, neither the control nor the test group showed any remarkable abnormality, and each organ was normal. Conclusions: It is thought that ingestion of ENM is useful for immunity enhancement.

• Journal of Food Hygiene and Safety
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• v.13 no.1
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• pp.6-13
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• 1998

This study was carried out to investigate the antioxidative activity on oxidaton of human low density lipoprotein from marine microorganisms. Bacillus sp. RH-5 producing antioxidative activity have been isolated and identified from coast sea in Pusan. Bacillus sp. RH-5 produced at highest level of antioxidative activity in the medium of 1.0% glucose, 0.25% polypepton, 0.25% yeast extract, 0.01% $FeSO_4{\cdot}7H_2O$ and 50% sea water. The optimal medium pH, cultural temperature and shaking time for the highest production as the antioxidant were 7.0, $30^{\circ}C$ and 48 hr, respectively. The culture broth inhibited the copper catalyzed oxidation of human low density lipoprotein (LDL) at the concentration of 500 and $1,000\;\mu\textrm{g}/ml$ ethylacetate extracts in the presence of $5\;\mu\textrm{M}\;CuSo_4$. The electrophoretic mobility of the LDL oxidized in the presense of $5\;\mu\textrm{M}\;CuSo_4$ was higher than that of native LDL.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.20-28
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• 1999

To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.