• Title/Summary/Keyword: Non-Isolation

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Isolation and identification of soycurd forming lactic acid bacteria which produce GABA from kimchi (김치로부터 GABA를 생산하는 커드 형성 젖산균의 분리 및 동정)

  • Kim, Eun-Ah;Mann, So-Yon;Kim, Su-In;Lee, Ga-Young;Hwang, Dae-Youn;Son, Hong-Joo;Lee, Chung-Yeol;Kim, Dong-Seob
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.705-711
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    • 2013
  • Gamma amino butyric acid (GABA), known as a non-protein amino acid and major inhibitory neurotransmitter in the brain, has several functional properties such as neurotransmission, induction of hypotension, tranquilizer, and diuretic effects. The purpose of this study was to isolate and identify lactic acid bacteria, producing high GABA in fermented soy curd. Thirty-two strains of tofu-forming lactic acid bacteria were isolated from kimchi which a traditional Korean food fermented with many kind of microorganism. Among 32 strains, four strains (strain No. 10, 104, 214, 249) formed firm soycurd. In order to select lactic acid bacteria having high GABA producing potential, the isolated strains were cultured in the soymilk and fermented for 48 hr at $37^{\circ}C$. A strain No. 383, which showed highest GABA contents in fermented soycurd, was identified as L. sakei by 16S rDNA sequencing and API analysis, and named as L. sakei 383. L. sakei 383 showed optimal growth up to 24 hr at $35^{\circ}C$ in MRS broth. The optimal time and temperature for GABA production were 18 hr and $35^{\circ}C$ in soymilk. In the optimal condition time and temperature, GABA content of fermented soycurd by L. sakei 383 was 8.65 mg/100 g.

Isolation of Microorganisms and Development of Microbial Augmentation for Treatment of Paper Mill Wastewater (제지폐수 처리용 미생물의 분리 및 복합 미생물제제의 개발)

  • Kang, Dae-Ook;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.554-560
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    • 2011
  • This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.

Isolation and Characterization of a Diesel-Degrading Bacterium, Gordonia sp. SD8 (디젤 분해 세균 Gordonia sp. SD8 분리 및 특성)

  • Hong, Sun-Hwa;Kim, Ji-Young;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.335-339
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    • 2010
  • A diesel-degrading bacterium, Gordonia sp. SD8, was isolated from soil contaminated with petroleum, and its diesel degradation was characterized in a soil as well as a liquid culture system. SD8 could grow in the mineral salt medium supplemented with diesel as a sole carbon and energy source. The maximum specific growth rate ($0.67{\pm}0.05\;d^{-1}$) and diesel degradation rate ($1,727{\pm}145$ mg-TPH $L^{-1}\;d^{-1}$) of SD8 showed at 20,000 mg-TPH $L^{-1}$ and $30^{\circ}C$, and then this bacterium could degrade high strength of diesel of 40,000 mg-TPH $L^{-1}$. The residual diesel concentration in the inoculated soil with SD8 was 3,724 mg-TPH kg-dry $soil^{-1}$ after 17 days, whereas the diesel concentration in the non-inoculated soil was $8,150{\pm}755$ mg-TPH kg-dry $soil^{-1}$. These results indicate that Gordonia sp. SD8 can serve as a promising microbial resource for the bioremediaion of contaminated soil with petroleum hydrocarbons including diesel.

Isolation, Identification, and Evaluation of Biocontrol Potentials of Rhizosphere Antagonists to Rhizoctonia solani (원예작물(園藝作物) 모잘록병(Rhizoctonia solani $K\"{u}hn$)의 발생(發生)에 관여하는 근권길항균(根圈拮抗菌)의 분리(分離), 동정(同定) 및 생물적(生物的) 방제(防除) 검토(檢討))

  • Kim, Hee-Kyu;Roh, Myung-Ju
    • Korean journal of applied entomology
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    • v.26 no.2 s.71
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    • pp.89-97
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    • 1987
  • Antagonistic microorganisms from rhizosphere soil were isolated, identified, and applied successfully as the biocontrol agents of damping-off caused by Rhizoctonia spp. Rhizosphere antagonists isolated from rhizosphere soil were identified as Trichoderma viride, T. harzianum, T. hamatum, T. polysporum, Gliocladium sp., Pseudomonas fluorescence, P. stutzeri, P. cepacia, Enterobacter sp., Serratia sp. and Erwinia herbicola. Of these, the most promising ones in vitro were T. virdie, T. harzianum, Gliocladium sp., Serratia sp., P. stutzeri, and P. cepacia. These above six antagonists were efficient in reducing disease incidence to $40{\sim}70%$ when the reselected rhizosphere antagonists preparations were applied to the soil at $10^6$ propagules per gram. Among six antagonists, T. viride was the most promising biocontrol agents against R. solani isolates in soil. The suppressive effect was more evident in steam-sterilized soil than in non-sterilized field soil.

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Antioxidative and Cytotoxicity Activities against Human Colon Cancer Cells Exhibited by Edible Crude Saponins from Soybean Cake (대두박 식용사포닌의 항산화 및 대장암세포 성장 억제효과)

  • Park, Kyung-Uk;Kim, Jae-Yong;Seo, Kwon-Il
    • Food Science and Preservation
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    • v.16 no.5
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    • pp.754-758
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    • 2009
  • To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.

Isolation of a Bacillus licheniformis DK42 Producing Cellulase and Xylanase, and Properties of the Enzymes (Cellulase 및 Xylanase를 분비하는 Bacillus licheniformis DK42의 분리 및 효소 특성)

  • Kim, Min Jeong;Lim, Soo Jin;Kang, Dae-Kyung
    • Journal of Animal Science and Technology
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    • v.50 no.3
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    • pp.429-436
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    • 2008
  • A bacterium producing cold-active cellulase and xylanase was isolated from pig feces. The isolate, DK42 strain, was found to be the Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus licheniformis DK42 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. The characterization of crude cellulase and xylanase from B. licheniformis DK42 was investigated. Cellulase exhibited an optimum temperature and pH at 45℃ and 6.0, whereas xylanase exhibited an optimum temperature and pH at 55℃ and 6.0. Especially cellulase maintained approx. 50% of its maximum activity even at 10℃, indicating that it is cold-active. Both cellulase and xylanase were stable after 2hr at 35℃, whereas they lost their activities after 30min at 65℃.

Purification and Properties of ${\beta}-Mannanases$ from Germinated Guar Bean (${\beta}-Mannanase$ 군(群)의 정제(精製) 및 그들의 성질(性質)에 관(關)한 연구(硏究))

  • Lee, Su-Rae
    • Applied Biological Chemistry
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    • v.7
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    • pp.1-13
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    • 1966
  • 1) Three ${\beta}-1$, 4-mannanases were isolated from germinated guar bean through extraction, ammonium sulfate fractionation, column chromatography on cellulose derivatives and gel filltration on Sephadex G-100. They were designated as ${\beta}-1$, 4-mannanase A,B and C, respectively, in the order of isolation. 2) These enzymes were different in several aspects such as pH optimum, effect of metal ions, adsorbability on cellulose derivatives, molecular weight, Michaelis constant toward reduced ivory nut mannan A, mode of action and extent of hydrolysis of the mannan. 3) ${\beta}-1$, 4-Mannanases A and C were proposed to be two different endo-enzymes of random-splitting type producing a series of oligosaccharides from ${\beta}-1$, 4-mannans. ${\beta}-1$, 4-Mannanase B was suggested to be possibly an exe-type enzyme catalyzing a stepwise splitting from the non-reducing end of ${\beta}-1$, 4-mannans to produce mannose. 4) Guaran was subjected to hydrolysis by the purified enzymes and the consequence was discussed in connection with structural requirements of the enzymes toward substituted ${\beta}-1$, 4-mannans and their role in germinating guar seeds.

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A Study on the Seismic Performance Design of Waterproofing Materials Applied Single-side Walls on Underground Structures (지하 구조물 외벽에 적용되는 방수재료의 구조체 거동 및 진동 대응 성능 설계 제안 연구)

  • Kim, Soo-Yeon;Kim, Meong-Ji;Oh, Sang-Keun
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.24 no.1
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    • pp.43-50
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    • 2020
  • In this study, the possibility of seismic performance design was proposed and the quality verification test method was reviewed as part of the design of waterproof performance in underground walls under accelerated environment conditions for waterproofing materials, which are barrier and finishing material that can prevent ingress or overflowing water from inside and outside of a building by attaching all of the construction materials used in construction structures. Considering the current state of earthquake-resistant design of construction materials in Korea and abroad, seismic product groups are rare and mostly dependent on construction methods because there are no regulations on materials, although there are still regulations on earthquake-resistant design in the building process under the current law. Overseas, it was possible to confirm that various building materials that gave seismic performance to non-structural materials, such as Japan, Canada, and Germany, are being developed. If it is possible to have a complementary response to earthquakes in the advanced external waterproofing materials, it can be expected to be applicable as leak prevention and prevention technology along with the seismic designed structure.

Isolation and purification of a cecropin-like antimicrobial peptide from the japanese oak silkworm, Antheraea yamamai (천잠 세크로핀 항균펩타이드 분리 및 정제)

  • Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho;Park, Seung-Won;Kim, Sung-Wan;Hwang, Jae-Sam;Kang, Seok-Woo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.145-149
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    • 2012
  • Cecropin is a well-studied antimicrobial peptide that play important role as key factor in insect humoral immunity. In this study, cecropin-like antimicrobial peptide was isolated and purified from the larval haemolymph of immune-challenged japanese oak silkworm, Antheraea yamamai. To isolate antimicrobial peptide, we separated and compared acidic extracted hemolymph protein bends between control and immune-challenged larvae using SDS-PAGE analysis. In the immune hemolymph extract, but not of non-immune hemolymph, we detected differential expressed peptide band with molecular mass 4,223.01 Da. To understand this peptide better, we successfully purified this peptide using cation exchange chromatography and gel permeation chromatography. Its N-terminal amino acid sequence obtained by Edman degradation evidenced a significant degree of identity with other lepidopteran cecropins. The purified A. yamamai cecropin-like peptide showed a broad spectrum of activity against fungi, Gram-negative and Gram-positive bacteria.

Isolation and Identification of Pheophytin, a Photosensitizer from Nostoc commune that Induces Apoptosis in Leukemia and Cancer Cells (Nostoc commune으로부터 백혈병세포와 간암세포에 대한 apoptosis 유도 광과민성물질 pheophytin a의 분리 및 구조동정)

  • Park, Jae-Eun;Lee, Jun-Young;Lee, Min-Woo;Jang, Eun-Jin;Hong, Chang-Oh;Kim, Keun Ki
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1321-1331
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    • 2018
  • The aim of this study was to separate the photosensitizer that induces apoptosis of U937 and SK-HEP-1 cells from Nostoc commune. Dried N. commune was extracted with $CH_2Cl_2/MeOH$ (1:1) to separate the photosensitizer using various chromatographic techniques. The isolated compound was identified as pheophytin a ($C_{55}H_{74}N_4O_5$) with a molecular weight of 870. Its photodynamic activities were assessed under different irradiation conditions (light and non-light) at the same concentration range of $1.15-23.0{\mu}M$. The apoptosis inducing activity in U937 or SK-HEP-1 cells appeared only in the light. The mechanisms underlying the pheophytin a-mediated photodynamic inhibition of cancer cells were further investigated by examining cell morphology changes, cytotoxicity, caspase-3/7 activity, fluorescence staining, flow cytometry analysis, and DNA fragmentation in these two cell lines. The positive control and the light irradiation group showed typical apoptotic responses, including morphological changes, cytotoxicity, caspase activity, nucleus shrinkage owing to chromatin condensation, DNA laddering, and the presence of apoptotic bodies. Cytotoxicity markedly increased in a dose-dependent manner after a 12 hr exposure. Caspase-3/7 activity was higher in U937 cells than in SK-HEP-1 cells. Apoptosis induction therefore appeared to be both concentration- and light-dependent. In conclusion, pheophytin a, isolated from the blue green alga N. commune, had a photodynamic apoptosis-inducing effect on U937 and SK-HEP-1 cells. The findings reported here can be used as basic data for the development of next-generation photosensitizers from N. commune.