• Journal of Microbiology and Biotechnology
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• v.30 no.5
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• pp.668-680
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• 2020

Bacillus velezensis is an important plant growth-promoting rhizobacterium with immense potential in agriculture development. In the present study, Bacillus velezensis Lle-9 was isolated from the bulbs of Lilium leucanthum. The isolated strain showed antifungal activities against plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea and Fusarium fujikuroi. The highest percentage of growth inhibition i.e., 68.56±2.35% was observed against Fusarium oxysporum followed by 63.12 ± 2.83%, 61.67 ± 3.39% and 55.82 ± 2.76% against Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several were identified as antimicrobial agents such as diketopiperazines, cyclo-peptides, linear peptides, latrunculin A, 5α-hydroxy-6-ketocholesterol, (R)-S-lactoylglutathione, triamterene, rubiadin, moxifloxacin, 9-hydroxy-5Z,7E,11Z,14Z-eicosatetraenoic acid, D-erythro-C18-Sphingosine, citrinin, and 2-arachidonoyllysophosphatidylcholine. The presence of these antimicrobial compounds in the bacterial culture might have contributed to the antifungal activities of the isolated B. velezensis Lle-9. The strain showed plant growth-promoting traits such as production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation and phosphate solubilization. IAA production was accelerated with application of exogenous tryptophan concentrations in the medium. Further, the lily plants upon inoculation with Lle-9 exhibited improved vegetative growth, more flowering shoots and longer roots than control plants under greenhouse condition. The isolated B. velezensis strain Lle-9 possessed broad-spectrum antifungal activities and multiple plant growth-promoting traits and thus may play an important role in promoting sustainable agriculture. This strain could be developed and applied in field experiments in order to promote plant growth and control disease pathogens.

• Applied Biological Chemistry
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• v.46 no.3
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• pp.165-170
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• 2003

The antioxidative activity of antioxidative substances produced from several bacterial strains isolated from fermented foods were tested by $DPPH\;({\alpha},{\alpha}'-diphenyl-{\beta}-picrylhydrazyl)$ free radical scavenging activity. One of the strains showing the highest antioxidative activity was identified as Bacillus sp. based on the morphological, biochemical, physiological characteristics, and 16S rRNA sequence, and named FF-7. The most optimal medium condition for the production of antioxidative substance from Bacillus sp. FF-7 was 2% galactose as carbon source and l% tryptone as nitrogen source. The antioxidative substance produced from FF-7 in these cultural medium was also tested by in vitro experimental models, the peruxidation of linoleic acid and the peroxidation of rat tissues microsomes by using thiobarbituric acid (TBA) for assay of free malondialdehyde production. The antioxidative activity against lipid peroxidation of rat tissues microsomes was shown in the following order; brain 97.50% > heart 79.95% > kidney 77.84% > spleen 77.47% > testis 69.96% > liver 62.45%. The antioxidative substance produced from FF-7 on linoleic acid peroxidation by IBA method was effectively inhibited during four days, and 0.05% BHT (butylated hydroxytoluene) used comparative control was also effectively inhibited. Results showed that the highest antioxidative activity by DPPH method of antioxidative substance produced from Bacillus sp. FF-7 was obtained by supplementing 2% galactose as carton source and l% tryptone as nitrogen source in cultured medium, this substance effectively inhibited the formation of TBARS in brain microsome in vit개 system and in linoleic acid peroxidation.

• Journal of Mushroom
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• v.12 no.1
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• pp.35-40
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• 2014

Several bacteria are known as the causal agents of diseases of the cultivated button mushroom(Agaricus bisporus) and oyster mushroom(Pleurotus ostreatus). Pseudomonas tolaasii is the causal agent of brown blotch disease of commercial mushrooms. Pseudomonas sp. HC1 is a potent biological control agent to control brown blotch disease caused by Pseudomonas tolaasii. This can markedly reduce the level of extracellular toxins (i.e., tolaasins) produced by Pseudomonas tolaasii, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the Pseudomonas sp. HC1, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 5.0 and $20^{\circ}C$, respectively. The optimal culture medium for the growth of tolaasin inhibitor bacterium was determined as follows: 0.9% dextrin, 1.5% yest extract, 0.5% $(NH_4)_2HPO_4$, 4mM $FeCl_3$, and 3.0% cysteine.

• Journal of Life Science
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• v.27 no.2
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• pp.211-216
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• 2017

This study aims to reduce the production cost of mushrooms and therefore boost the income of mushroom producers. The addition of a 10% concentration of herb medicine refuse was adequate for promoting mycelium growth in Flammulina velutipes. A moisture content of 60% in the culture medium was adequate for promoting hyphal growth. The optimum temperature and humidity were $16^{\circ}C$ and 70%-85%, respectively. The average yield of Flammulina velutipes fruting body per 1,100 ml bottle was 275-282 g, which is similar to that of the control. The outbreak of diseases such as mildew and bacterial blotch in mushroom was never observed in the bottle treated with herb medicine refuse. Fewer deformed and second-class quality mushrooms were observed than in the control group. Deviation reduced, and mushroom growth was observed to be significantly uniform. The winter mushroom had a white fruiting body with a hemispherical shape. The diameter of the mushroom cap was 9.2-9.3 mm, stipe length was 12.7-12.8 cm, and thickness was 3.3-3.4 mm; these were almost the same as those in the control group. The partial hardness of winter mushrooms in both the experimental and the control groups showed that the stipe ($54.6-57.3g/cm^2$) is a little harder than the mushroom cap ($46.8-47.6g/cm^2$). The calories and nutrients per 100 g of mushrooms in the control and samples were similar to each other.

• KSBB Journal
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• v.17 no.6
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• pp.571-576
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• 2002

Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

• Journal of Life Science
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• v.17 no.7 s.87
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• pp.970-975
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• 2007

For the screening of ${\gamma}-aminobutyric$ acid (CABA)-producing bacteria, 86 bacterial strains which produce GABA were isolated from Kimchi and Salted fisk .Among these, three strains designated AML15, AML45-1, AML72 with relatively high GABA productivity were selecled by thin layer chromatography (TLC). To elucidate the relationship between isolated strains and the genus Lactobacillus, their 16S rDNA sequence were examined. The result of their DNA sequences showed 99% similarity with Lactobacillus brevis ATCC 367. On the basis of the these results, isolated strains were identified as Lactobacillus brevis and designated L. brevis AML15. In order to determine the optimum conditions for GABA production, the isolated strains were cultivated in pyridoxal phosphate (PLP) and monosodium glutami. acid (MSG). Results showed that L. brevis AML15 had the highest CABA productivity with 10,424 $nM/{\mu}l$ concentration in MRS broth containing 5% (w/v) MSG and 10 ${\mu}M$ PLP at pH 5.0. The results imply that L. brevis AML15 has the potential to be developed as a strain for GABA hyper-production.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.157-161
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• 2010

Photosynthetic purple nonsulfur bacterial strains were isolated from the sediments collected from rice paddy fields and sludges of wastewater treatment plant, and their plant growth promoting capabilities were examined. Most well known phytohormones, auxin such as indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) and 5'-aminolevulinic acid (ALA) were detected by HPLC in the culture broth of these isolates. Among the isolated bacteria, Rhodopseudomonas faecalis D15 showed the highest production rate of 769.8 ${\mu}g$/mg protein of IAA, 1323 ${\mu}g$/mg protein of IBA and 7.4 mM/mg protein of ALA in the modified Biebl and Pfennig's medium. R. faecalis C9 showed the highest production rate of 20.82 ${\mu}g$/mg protein of gibberellin. In consequence, the root length and dry weight of the germinated tomato seedling treated with R. faecalis isolates were longer and heavier than those of uninoculated control after 15 days of incubation in the soil. Especially, the dry weight of germinated tomato seedling increased by 119.4% in C9-treated samples after 15 days. These purple nonsulfur bacteria may be utilized as environment-friendly biofertilizer in the agriculture.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.460-465
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• 1996

The production of bacterial protease and its characteristics were investigated with Bacillus subtilis globigii CCKS-118 which was isolated from Korean traditional soy sauce. The optimum culture condition of the strain for the production of alkaline protease was as follow : 2% soluble starch, 0.2% yeast extract, 0.1% $(NH_4)_2SO_4$, 0.2% $MgSO_4$, pH 7.5, $35^{\circ}C$ and 20h rs. The optimum pH and temperature for the enzyme action of alkaline protease producing Bacillus subtilis globigii CCKS-118 were pH 9.0 and $50^{\circ}C$, respectively. The enzyme was relatively stable at $pH\;6.0{\sim}9.0$ and at temperature below $40^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{2+}$ whereas $Cu^{2+}$ gave rather activating effects on the enzyme activity. The enzyme was inhibited by phenylmethane-sulfonyl fluoride indicating serine pretense metal ion group are required for the enzyme activity. Km value was $1.242{\times}10^{-4}M$, $V_{max}$ value was $25.99\;{\mu}g/min$. This enzyme hydrolyzed casein more rapidly than the hemoglobin.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.657-663
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• 2005

Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

• Journal of Society of Preventive Korean Medicine
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• v.16 no.3
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• pp.139-153
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• 2012

Objectives : NK cells are spontaneously cytotoxic lymphocytes. These are not only important parts in the first line of defence against bacterial and viral infections of outside, but they may also play a critical role in chronic viral diseases. NK cells kill their targets spontaneously, without the need for prior sensitization and class I MHC restriction by the regulation of cytolytic functions and secretion of a variety of cytokines, such as interleukin-12(IL-12), MCP-1, IL-6, TNF-${\alpha}$, IFN-${\gamma}$. In addition, macrophage and NK cells cooperate through the production of cell mediates. These cooperation and modulation are one of major factors to prevent for evading immune surveillance of cancer. Hence, it could be assumed that if any candidate to enhance activities of macrophage and NK cell, it is considered as a potentially useful agents against cancer. Methods : In our study, to investigate effect of fermented soybean extracts by Lactic acid bacteria (SFE, soybean fermented extracts) work on intestinal immune cell to maintain general immune modulating and anti-cancer activity. We analyzed NK cytotoxicity assay and gene expressions of cytokine related with macrophage and NK cell activity. Results : In vitro experiment, SFE was verified as safety material for cell toxicicty to tumor cell strain without any toxicity of tumor growth inhibition and various cell strain. Effects of macrophage activity stimulating directly by SFE measured induced cytokine. The studies showed that IL-12 production by stimulation of SFE depended on concentration from 0.16mg/mL to 0.63mg/mL with non toxicity to cell, and it was the best activity at 0.63mg/mL. Besides, the effective concentration of SFE producing TNF-${\alpha}$ is similar to IL-12, but it was the best activity at 1.25mg/mL. The level of MCP-1, IL-6 and IFN-${\gamma}$ depended on concentration from 0.16mg/mL to 10mg/mL, IFN-${\gamma}$ showed the best activity at the effective concentration of 0.63mg/mL. With the result of NK cell activity measurement, the spleen cell of mouse injected SFE had 1.5 times higher killing effect than non injected cell. Conclusions : The result of this studies is that Soybean fermetated extracts(SFE) has possibility to immune aided material for the function not only inhibition of microbial infection to macrophage but also activity of adaption immune and cellular immune system.