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Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Transcriptome Analyses for the Anti-Adipogenic Mechanism of an Herbal Composition (생약복합물의 지방세포형성억제 기전규명을 위한 전사체 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1054-1065
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    • 2010
  • SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.

Therapeutic Effect of Toothpaste Containing Hydroxyapatite and Tribasic Calcium Phosphate on Dentinal Hypersensitivity (치아 과민증에 Hydroxyapatite와 Tricalcium phosphate을 함유한 치약의 치료효과)

  • Choi, Yea Hun;Park, Hyean Cheal;Lee, Sang Mong;Son, Hong Joo;Choi, Eun Bi;Ha, Jun Young;Lee, Jun Young;Kim, Keun Ki
    • Journal of Life Science
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    • v.24 no.6
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    • pp.642-647
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    • 2014
  • Although it is not a pathological symptom, Dentinal Hypersensitivity (DH) describes pain felt by patients whose tooth roots are exposed outside of the gums and are therefore sensitive to external stimuli. DH is caused by tooth brushing or gum diseases and treatment to reduce the sensitivity can include use of materials having stimulation activity for DH or a resin material applied periodontally. This study examined the hypersensitivity treatment effects of a four-week treatment with a toothpaste containing hydroxyapatite and tricalcium phosphate (Hap-TCP toothpaste). The Hap-TCP toothpaste was made by mixing a commercially available fluorine-containing toothpaste with 10% (W/W) hydroxyapatite and 19% (W/W) tricalcium phosphate (both 99% purity based on XRD analysis). The tooth hypersensitivity treatment effect was surveyed by scoring VRS values, and showed no significant initial difference compared with the control. However, after 1 week of use, the pain reduction value was 8% in the treatment group compared to the control group. This value increased to 30% and 60% after 2 and 4 weeks, respectively. Hypersensitivity to cold stimulation, which was used as a VAS value, showed no initial significant differences compared with the control, but was significantly decreased after 1, 2, and 4 weeks in the experimental group, with more than a 3-fold difference after 4 weeks. These findings confirmed that remineralization can alleviate DH as hydroxyapatite fills dentinal tubules and calcium, phosphorus, and tricalcium phosphate ion equilibrium is established.

Impact of Elevated Temperature and CO2 on Growth and Fruit Quality of Pepper (Capsicum annuum L.) (온도 및 CO2 상승이 고추의 생육 및 과실품질에 미치는 영향)

  • Song, Eun Young;Moon, Kyung Hwan;Son, In Chang;Wi, Seung Hwan;Kim, Chun Hwan;Lim, Chan Kyu;Oh, Soonja
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.18 no.4
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    • pp.179-187
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    • 2016
  • This study was conducted to determine the impact of elevated temperature and $CO_2$ concentration based on climate change scenario on growth and fruit quality of pepper (Capsicum annuum L. cv. Muhanjilju) with SPAR (Soil Plant Atmosphere Research) chamber. The intraday temperatures of climate normal years fixed by $20.8^{\circ}C$ during the growing season (May 1~October 30) of climatic normal years (1971~2000) in Andong region. There were treated with 4 groups such like a control group (ambient temperature and 400ppm $CO_2$), an elevated $CO_2$ group (ambient temperature and 800ppm $CO_2$), an elevated temperature group (ambient temperature+$6^{\circ}C$ and 400ppm $CO_2$) and an elevated temperature/$CO_2$ group (ambient temperature+$6^{\circ}C$ and 800ppm $CO_2$). Compared with the control, plant height, branch number and leaf number increased under the elevated temperature and elevated temperature/$CO_2$ group. However, leaf area and chlorophyll content showed a tendency of decreasing in the elevated temperature group and elevated temperature/$CO_2$ group. The number of flower and bud were decreased in the elevated temperature and elevated temperature/$CO_2$ group (mean temperature at $26.8^{\circ}C$) during the growth period. The total number and the weight of fruits were decreased in the elevated temperature group and elevated temperature/$CO_2$ group more than the control group. While the weight, length and diameter of fruit decreased more than those of control as the temperature and $CO_2$ concentration increased gradually. This result suggests that the fruit yield could be decreased under the elevated temperature/$CO_2$ ($6^{\circ}C$ higher than atmospheric temperature/2-fold higher than atmospheric $CO_2$ concentration), whereas the percentage of ripen fruits after 100 days of planting was increased, and showed earlier harvest time than the control.

Photosynthetic and Growth Responses of Chinese Cabbage to Rising Atmospheric CO2 (대기 중 CO2 농도의 상승에 대한 배추의 광합성과 생장 반응)

  • Oh, Soonja;Son, In-Chang;Wi, Seung Hwan;Song, Eun Young;Koh, Seok Chan
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.18 no.4
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    • pp.357-365
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    • 2016
  • The effects of elevated atmospheric $CO_2$ on photosynthesis and growth of Chinese cabbage (Brassica campestris subsp. napus var. pekinensis) were investigated to predict productivity in highland cropping in an environment where $CO_2$ levels are increasing. Vegetative growth, based on fresh weight of the aerial part, and leaf characteristics (number, area, length, and width) of Chinese cabbage grown for 5 weeks, increased significantly under elevated $CO_2$ ($800{\mu}mol{\cdot}mol^{-1}$) compared to ambient $CO_2$ ($400{\mu}mol{\cdot}mol^{-1}$). The photosynthetic rate (A), stomatal conductance ($g_s$), and water use efficiency (WUE) increased, although the transpiration rate (E) decreased, under elevated atmospheric $CO_2$. The photosynthetic light-response parameters, the maximum photosynthetic rate ($A_{max}$) and apparent quantum yield (${\varphi}$), were higher at elevated $CO_2$ than at ambient $CO_2$, while the light compensation point ($Q_{comp}$) was lower at elevated $CO_2$. In particular, the maximum photosynthetic rate ($A_{max}$) was higher at elevated $CO_2$ by 2.2-fold than at ambient $CO_2$. However, the photosynthetic $CO_2$-response parameters such as light respiration rate ($R_p$), maximum Rubisco carboxylation efficiency ($V_{cmax}$), and $CO_2$ compensation point (CCP) were less responsive to elevated $CO_2$ relative to the light-response parameters. The photochemical efficiency parameters ($F_v/F_m$, $F_v/F_o$) of PSII were not significantly affected by elevated $CO_2$, suggesting that elevated atmospheric $CO_2$ will not reduce the photosynthetic efficiency of Chinese cabbage in highland cropping. The optimal temperature for photosynthesis shifted significantly by about $2^{\circ}C$ under elevated $CO_2$. Above the optimal temperature, the photosynthetic rate (A) decreased and the dark respiration rate ($R_d$) increased as the temperature increased. These findings indicate that future increases in $CO_2$ will favor the growth of Chinese cabbage on highland cropping, and its productivity will increase due to the increase in photosynthetic affinity for light rather than $CO_2$.

Antioxidative Activity of Hot Water Extracts from Medicinal Plants (한약재 열수추출물의 항산화 활성)

  • Ju, Jong-Chan;Shin, Jung-Hye;Lee, Soo-Jung;Cho, Hee-Sook;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.7-14
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    • 2006
  • The antioxidative activity and the related parameters of hot water extracts obtained from 16 medicinal plants were tested. The extraction yield was the highest in Salvia miltiorrhiza Bunge $(36.49\%)$ pH range was $4.00\~5.92$ in all samples. Absorbance at 280 nm was examined to determine aromatic compounds content. The absorbance of $250{\mu}g/mL$ sample was the highest in Prunella vulgaris Linne var. lilacina Nakai (2.872) and below 0.5 in Cirsium maackii, Crataegus pinnatifida Bunge and Zizyphus jujuba Miller. Also, absorbance at 420 nm was high in order of Prunella vulgaris Linne vu. lilacina Nakai (1.312), Zea mays Linne (0.917) and Inula japonica Thunberg (0.725) Total phenolic compounds contents was the highest in Prunella vulgaris Linne var. lilacina Nakai $(5.07{\pm}0.05\;mg/100g)$ and flavonoids contents was 2-fold higher in Salvia miltiorrhiza Bunge $(4.82{\pm}1.16\;mg/100g)$ than the other samples. Electron donating abilities of Zizyphus jujuba Miller, Cornus officinalis Siebold et Zuccarini and Salvia miltiorrhiza Bunge were over $90\%$ at $1000{\mu}g/mL$. Reducing power had similar tendency to electron donating ability while reducing power was significantly lower in samples compared to BHT. Two samples (Prunella vulgaris Linne val. lilacina Nakai and Inula japonica Thunberg) were found to have more than $50\%$ nitrite scavenging effect at $500{\mu}g/mL$ while 8 kinds samples (Zizyphus jujuba Miller, Cornus officinalis Siebold 81 Zuccarini, Chrysanthemum indicum Linne, Prunella vulgaris Linne var. lilacina Nakai, Inula japonica Thunberg, Acanthopanax sessiliflorum Seeman, Salvia miltiorrhiza Bunge and Curcuma longa Linne) showed more than $50\%$ nitrite scavenging effect at $1000{\mu}g/mL$. Prunella vulgaris Linne var. lilacina Nakai showed significantly stronger nitrite scavenging effect than other samples, and its activity was $59.62{\pm}1.573\%$ and $80.58{\pm}0.300\%$ at concentrations of 250 and $1000{\mu}g/mL$, respectively.

Antibacterial Activities of Trace Elements in Combination with Food Additives (미량원소 강화 식품소재의 항균효과)

  • Kim, Bo-Mi;Mok, Jong-Soo;Oh, Eun-Gyoung;Son, Kwang-Tae;Shim, Kil-Bo;Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.35-41
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    • 2006
  • Antibacterial activities of the trace elements in combination with the food additives were measured against 6 kinds of food-borne microorganisms such as Escherichia coli, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis and Pseudomonas fluorescens. The difference of antibacterial activity was not shown among the kinds of food additives, such as dextrin, gelatin and collagen. Zn and Ge in combination with food additives had strong antibacterial effect. Especially, $1\%$ zinc acetate in combination with $1\%$ gelatin was more effective against P. fluorescens and S. aureus than against Bacillus sp., E. coli and V. parahaemolyticus. Minimum inhibitory concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on S. aureus and P. fluorescens, and 1.0 mg/mL on E. coli, V. parahaemolyticus, B. cereus and B. subtilis. Minimum bactericidal concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on P. fluorescens and 1.0 mg/mL on E. coli, V. parahaemolyticus, S. aureus, B. cereus and B. subtilis. The zinc acetate in combination with gelatin showed stronger inhibitory effect in acidic range below pH 6.0, and remained active even after heat treatment at $121^{\circ}C$ for 15 min. In comparison with control, the viable cell counts of fish pastes, which were coated with the solution containing both $1\%$ zinc acetate and $3\%$ gelatin, were decreased by more than 100-fold until the storage of 7 days at $10^{\circ}C$. The results indicate that the combined use of zinc acetate and some food additives could prolong the shelf life of fish pastes by 8 days or more at $10^{\circ}C$.

A Study on the Antioxidative, Anti-inflammatory and Anti-thrombogenic Effects of Zanthoxylum piperitum DC. Extract (초피나무 추출물의 항산화, 항염증 및 항혈전 효능에 관한 연구)

  • Jang, Mi-Jin;Rhee, Soon-Jae;Cho, Sung-Hee;Woo, Mi-Hee;Choi, Jeong-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.21-27
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    • 2006
  • Effects of root, stem and leaf extracts of Zanthoxylum piperitum on the inhibition of lipid peroxidation in the hepatic microsome of rat, DPPH radical scavenging activity, soybean lipoxygenase activity and activated partial thromboplastin times (APTT) were examined in vitro. The highest inhibition of hepatic microsomal lipid peroxidation was observed by ethyl acetate fraction of the root and stem extracts. The high inhibition of lipid peroxidation was observed in the leaf, the root and the stem in order. The DPPH radical scavenging activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the steam extract. It was similar to the inhibition of hepatic microsomal lipid peroxidation. The DPPH radical scavenging activity was the highest in 0.50 mg/mL of ethyl acetate fraction, and it was 4.4-fold higher than that of a-tocopherol, as an antioxidant standard. The DPPH radical scavenging activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The soybean lipoxygenase activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the stem extracts. The soybean lipoxygenase activity was the highest in 0.50 mg/mL of ethyl acetate fraction. The soybean lipoxygenase activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The leaf extract showed the highest antithrombogenic effect followed by the stem and then the root extract. The activated partial thromboplastin times were dependent on the extract concentration in the range of $0.10\~2.00$ mg/mL.

Effects of Dietary Vitamin E and Selenium on Hematopoiesis and Antioxidative Detoxification Mechanism in Lead Poisoned Rats (식이 Vitamin E와 Selenium이 납중독된 흰쥐에 있어서 조혈작용과 항산화적 해독기구에 미치는 영향)

  • 이순재;박규영;김관유
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.651-657
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    • 1993
  • The protective effects of dietary vatamin E and selenium on peroxidative damage and hematopoietic inhibition by lead poisoning were investigated in rats. Male Sprague-Dawley rats weighing 150$\pm$5g were divided into six groups according to dietary vitamin E and / or selenium levels, i.e. control(vitamin E, 40mg/kg diet), 0E(without vitamin E, Se), 40E(vitamin E, 40mg/kg diet ; without Se), 200E(vitamin E, 200mg/kg diet ; without Se), 200ES(vitamin E, 200mg/kg diet ; Se, 0.5ppm) and 0Es(without vitamin E ; Se, 0.5ppm) groups. All experimental groups were fed ad libitum 2000ppm lead in diet except control for 4 weeks. Hemoglobin contents and hematocrit values of lead groups were lower than control group except 200ES group and were the lowest in 0E group. Aminolevulinic acid dehydratase(ALAD) activities of blood and liver were sequentially reduced in 200ES, 200E, 0ES, 40E and 0E groups, compared to control, were as urinary aminolevulinic acid (ALA) excretions were increased in the groups which represented low ALAD activity. Heapatic superoxide dismutase(SOD) activities was lower in 0E, and higher in 40E, 200E and 200ES groups, compared with control. Glutathione peroxidase(GPX) activities of liver were reduced in 0E and 40E groups, but those of 0ES, 200E and 200ES groups were significantly increased. Especially GPX activities in 200ES and 200ES groups were not different from control group. The reduced glutathione contents in liver were lowest in 0E and 40E groups, compared with control, whereas levels of the oxidized form were opposite phenomena of that. Liver lipid peroxide values of 0E, 0ES, 40E and 200E groups were 6.4, 2.9, 2.1 and 1.3 fold higher than control, respectively, but 200ES groups was not different from control.

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Enhanced production of monacolin-K through supplement of monacolin-K precursors into production medium and cloning of SAM synthetase gene (metK) (Precursor제공 및 생합성 관련 유전자의 cloning을 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.23 no.6
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    • pp.519-524
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    • 2008
  • Monacolin-K is a strong anti-hypercholesterolemic agent produced by Monascus sp. via polyketide pathway. High-yielding mutants of monacolin-K were developed through rational screening strategies adopted based on understanding of monacolin-K biosynthetic pathway. Through the experiments for investigating various amino acids as putative precursors for the monacolin-K biosynthesis, it was found that production level of monacolin-K was remarkably increased when optimum amount of cysteine was supplemented into the production medium. We suggested that these phenomena might be related to the special roles of SAM (S-adenosyl methionine), a putative methyl group donor in the biosynthetic pathway of monacolin-K, demonstrating close interrelationship between SAM-synthesizing primary metabolism and monacolin-K synthesizing secondary metabolism. Namely, increase in the intracellular amount of SAM derived from the putative precursor, cysteine which was extracellularly supplemented into the production medium might contribute to the significant enhancement in the monacolin-K biosynthetic capability of the highly mutated producers. On the basis of these assumptions derived from the above fermentation results, we decided to construct efficient expression vectors harboring SAM synthetase gene (metK) cloned from A. nidulans, with the hope that increased intracellular level of SAM could lead to further enhancement in the monacolin-K production through overcoming a rate-limiting step associated with monacolin-K biosynthesis. Hence, in order to overcome the plausible rate-limiting step associated with monacolin-K biosynthesis by increasing intracellular level of SAM, we transformed the producer mutants with an efficient expression vector harboring gpdA promoter of the producer microorganism, and metK gene. Notably, from the resulting various transformants, we were able to screen a very high-yielding transformant which showed approximately 3.3 fold higher monacolin-K productivity than the parallel nontransformed mutants in shake flask cultures performed under the identical fermentation conditions.