• Title/Summary/Keyword: 약 배양

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Purification and Gene Analysis of Peptidyl Prolyl cia-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus Peptidyl Prolyl cis-trans Isomerase의 정제 및 유전자 분석)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.104-111
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    • 2002
  • The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.

Characterization of Extracellular Cholesterol Oxidase Produced from Soil Microorganism (토양 미생물로부터 생산된 Extracellular Cholesterol Oxidase의 특성)

  • Park, Jeong-Su;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1507-1514
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    • 2008
  • Cholesterol oxidase catalyses the conversion of cholesterol to 4-cholesten-3-one. This enzyme has been used for clinical assay of human serum cholesterol and for reduction of cholesterol level in foods and feeds. In order to search the microorganism which has a high extracellular and stable activity of cholesterol oxidase, soil microorganisms were screened. As a result, the one with the highest extracellular cholesterol oxidase activity was obtained and named as the BEN 115. The BEN 115 strain was identified as one of the Nocardia species based on our taxonomic studies. The cholesterol oxidase from this strain was shown to have two bands of extracellular proteins on SDS-PAGE and Western blot. Their molecular masses were estimated to be about 55 and 57 kDa, respectively. In addition, this cholesterol oxidase was considerably stable at the broad range of pH $3.5{\sim}9.5$ and at the temperature of $25{\sim}55^{\circ}C$. The optimum pH and temperature of this cholesterol oxidase were pH 5.5 and $35^{\circ}C$, respectively. The activity of extracellular cholesterol oxidase could be enhanced 1.6 to 2.0 folds by the addition of nonionic detergent such as Triton X-114, Triton X-100, or Tween-80 into the culturing broth. The substrate specificities against campesterol, sitosterol and stigmasterol were measured to be 50%, 50%, and 27%, respectively, compared to the cholesterol. These results suggest that Nocardia sp. BEN 115 may be useful as a microbial source of cholesterol oxidase production.

Effects of Artemisia Capillaris Thunberg on Apoptosis in HeLa Cells (사철쑥의 HeLa 세포고사 효과)

  • Lee, Hyoung-Ja;Kim, Kee-Hwan;Park, Jong-Kun;Hwang, Eun-Hee
    • Journal of Nutrition and Health
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    • v.41 no.1
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    • pp.22-30
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    • 2008
  • This study analyzes the apoptosis of HeLa cells to see if we can use the Artemisia capillaris Thunberg for the prevention of chronic degenerative diseases. We used the HeLa cells to see what effects the A. capillaris Thunberg had on apoptosis of the cancer cells. We checked the cell activity, cell morphological change, DNA fragmentation, and DNA content after administering 0, 100, 500, 1000, and $2000{\mu}g/ml$ methanol, ethyl acetate, n-butanol extract of the A. capillaris Thunberg. As for the cell viability, the increase of concentration of methanol and ethyl acetate decreased the survival rate of the cell, but the phenomenon was much weakened in n-butanol extract and was not observed in aqueous extract. The higher the density of the methanol, ethyl acetate, n-butanol and aqueous extract was, the lower the survival rate of the HeLa cell was. These extracts obstructed the cell cohesion and caused the blebbing of he cell membrane and fragmentation of the nucleus, both of which are symptoms of apoptosis. Laddering-pattern DNA fragmentation was observed in the groups that were treated with the $1000{\mu}g/ml$ and $2000{\mu}g/ml$ of methanol extract. The DNA content of the cells apoptosis measured by fluorescent-activated cell sorter (FACS) increased as the density of the methanol, ethyl acetate and butanol extract increased. The result of the study shows that A. capillaris Thunberg fosters the apoptosis of HeLa cells, which suggests that the A. capillaris Thunberg has a great potential value as food additives, medicinal supplements for patients with chronic diseases, and preventive measures against cancer.

Cellular Responses of the TNT-degrading Bacterium, Stenotrophomonas sp. OK-5 to Explosive 2,4,6-Trinitrotoluene (TNT) (폭약 2,4,6-Trinitrotoluene에 노출된 분해세균 Stenotrophomonas sp. OK-5의 세포반응)

  • 장효원;송승열;김승일;강형일;오계헌*
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.247-253
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    • 2002
  • The cellular responses of TNT-degrading bacterium, Stenotrophomonas sp. OK-5 to explosive 2,4,6-trini-trotoluene (TNT) as an environmental contaminant were examined. Survival of the strain OK-5 with time in the presence of different concentrations of TNT under sublethal conditions was monitored, and viable counts paralleled the production of the stress shock proteins in this bacterium. Total cellular fatty acids analysis showed that strain OK-5 produced or disappeared several different kinds of lipids when grown on TNT media than when grown on TSA. Under scanning electron microscope, the cells treated with 0.5 mM TNT for 12 hrs showed irregular rod shapes with wrinkled surfaces. Analyses of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that several stress shock proteins including 70 kDa DnaK and 60 kDa GroEL in strain OK-5 were newly synthesized at different TNT concentrations in exponentially growing cultures. 2-D PAGE of soluble protein fractions from the culture of OK-5 exposed to TNT demonstrated that approximately 300 spots were observed on the silver stained gel ranging from pH 3 to pH 10. Among them, 10 spots significantly induced and expressed in response to TNT were selected and analyzed. As the result of internal amino acid sequencing with ESI-Q TOF, two proteins, spot #1 and spot #10 were assigned the DnaK protein XF2340 of Xylella fastidiosa and stress-induced protein of Mesorhizobium loti, respectively.

Isolation and Identification of Halotolerant Bacillus sp. SJ-10 and Characterization of Its Extracellular Protease (세포외 Protease를 생산하는 내염성 Bacillus sp. SJ-10 균주의 분리 동정 및 효소 특성)

  • Kim, Eun-Young;Kim, Dong-Gyun;Kim, Yu-Ri;Choi, Sun-Young;Kong, In-Soo
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.193-199
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    • 2009
  • A bacterium producing the halotolerant extracellular protease was isolated from squid jeotgal, and was identified as Bacillus sp. SJ-10 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The strain grew at $20^{\circ}C\sim55^{\circ}C$, pH 5~8, and 0%~14% NaCl and optimal growth conditions were $35{\pm}5^{\circ}C$, pH 7, and 5% NaCl. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and $C_{16:0}$ DNA G+C content was 50.58 mol% and menaquinone consisted of MK-7 Phylogenic analysis based on the 16S rRNA gene sequence indicated that SJ-10T belongs to the genus Bacillus. About 40 kDa of the salt-tolerant protease was purified by 40% ammonium sulfate saturation and Mono Q column chromatography. The optimal activity of the protease was pH 8 and stable at pH 5~10. The optimum temperature and NaCl concentration were $35{\pm}5^{\circ}C$ and $5{\pm}1%$, respectively.

Dissolved Oxygen at the Bottom Boundary Layer of the Ulleung Basin, East Sea (동해 울릉분지 해저 경계면의 용존산소)

  • Kang, Dong-Jin;Kim, Yun-Bae;Kim, Kyung-Ryul
    • Ocean and Polar Research
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    • v.32 no.4
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    • pp.439-448
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    • 2010
  • General consensus on typical vertical profile of dissolved oxygen in the Ulleung Basin is that dissolved oxygen concentration beyond 300 m decreases with increasing depth. However, the results of our observations in 2005 and 2006 revealed three different dissolved oxygen distribution types in the deep layer of the Ulleung Basin. The first type showed oxygen concentration decreasing with increasing depth (Type-1), the second showed oxygen concentration decreasing very sharply near the bottom boundary layer but constant in the bottom adiabatic layer (Type-2), the final was of the oxygen minimum layer above the bottom boundary layer (Type-3). Type-2 was the most common pattern in the Ulleung Basin. Type-1 was most common close to the Japan Basin, including the Ulleung Interplane Gap, while Type-3 was found around Dok do. Oxygen Consumption Rate (OCR) at surface sediment estimated using the dissolved oxygen distribution at the bottom boundary layer was $0.2{\sim}5.8\;mmol{\cdot}m^{-2}d^{-1}$, which coincided with OCR from direct sediment incubation. This implies that organic matter decomposition at surface sediment may play an important role in dissolved oxygen distribution patterns at the bottom boundary layer of the Ulleung Basin.

Conservation Environment for Mural Tomb in Goa-ri, Goryeong (고령 고아리 벽화고분의 보존환경 연구)

  • Jeong, Seon Hye;Lee, Hyun Ju;Lee, Min Young;Chung, Yong Jae
    • Journal of Conservation Science
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    • v.33 no.3
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    • pp.189-201
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    • 2017
  • This study focused on the Gaya mural tomb located in Goa-ri, Goryeong-gun, North Gyeongsang Province, Republic of Korea. Environmental factors such as ambient temperature, relative humidity and surface temperature, and microbial distribution on mural tombs were monitored for one year to gather data on the conservation environment of the mural tombs. The average internal temperature difference with reference to the ambient outer temperature was observed to be $11.7^{\circ}C$ for the monitoring period and the internal temperature of the tomb was found to change periodically every one or two months in response to the outer temperature. The highest temperature was observed in September and the lowest in March. The relative humidity in the mural tomb remained constant at 100%. Between December and April, condensation occurred on the ceiling of the main room of the tomb, where the murals are located. On the ceiling of the aisle, the condensation occurred throughout the year. The inside and surface wall were isolated from microorganisms, which could grow when a suitable growth environment suitable is established. Based on microbial growth temperature conditions, risk periods of microbial hazards were established, where in the period from August to October was identified as the most dangerous.

Cultural Conditions of Lactobacillus sp. GM7311 for the Production of Bacteriocin (Lactobacillus sp. GM7311에 의한 박테리오신의 생산 조건)

  • LEE Myung Suk;CHANG Dong Suck;KANG Ji-Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.5
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    • pp.834-841
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    • 1997
  • A lactic acid bacteria which showed antimicrobial activity was isolated from dairy products and identified as Lactobacillus sp. according to the morphological, physiological and biochemical properties, which was named Lactobacillus sp. SH 7311. The bacteriocin of Lactobacillus sp. GM 7311 showed a broad range of inhibitory spectrum against some gram positive and negative bacteria. Especially, Proteus mirabilis was highly sensitive to bacteriocin and used as indicator strain for further investigation. The optimal condition for the production of bacteriocin was showed on MRS broth at $37^{\circ}C$ and pM 6.0. Bacteriocin production of this strain cultured under optimal condition was increased late logarithmic phase to early stationary phase. This bacteriocin was fully active at the pH range $2.0\~5.0$, also was stable at $100^{\circ}C$ for 60 min. at pH 5.0, But about $40\%$ of bacteriocin activity was diminished by the treatment of acetone, ethanol, iso-butanol and ethyl ether during 2 hours at $4^{\circ}C$.

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Inherent Optical Properties of Red Tide Algal for Ocean Color Remote Sensing Application (해색원격탐사 활용을 위한 적조생물종 고유 광특성 연구)

  • Ahn, Yu-Hwan;Moon, Jeong-Eon;Seo, Won-Chan;Yoon, Hong-Joo
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.12 no.1
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    • pp.47-54
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    • 2009
  • This research is about the inherent optical properties(IOPs) of algae which is collected from Nam-Hae for basic research of red tide remote sensing technique development. 21 kinds of red tide organisms were cultivated to investigate IOPs of them in the level of laboratory, and specific absorption coefficient of phytoplankton($a^*$) and backscattering coefficient of phytoplankton(${b_b}^*$) are estimated by using spectrophotometer. Absorption spectrums according to species appeared to range from 0.005 to 0.06 ($m^2{\cdot}mg^{-1}$), and the shapes of spectrums were also different. The range of ${b_b}^*$ appeared to be $10^{-2}{\sim}10^{-4}\;m^2{\cdot}mg^{-1}$, which had about 100 times differences between species, and the shape of spectrum have significant difference between species. These results will input as a remote sensing reflectance model input parameter from ocean color.

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Selection of Microalgae for Advanced Treatment of Swine Wastewater and Optimization of Treatment Condition. (축산폐수의 3차 처리를 위한 미세조류의 선별 및 처리조건의 최적화)

  • 김성빈;이석준;김치경;권기석;윤병대;오희목
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.76-82
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    • 1998
  • The feasibility of algae as means of removing nitrogen and phosphorus from secondary treated swine wastewater was studied. Among the tested 7 species of Chlorella vulgaris (UTEX 265), Chlorella sp. GE 21, Botryococcus braunii (UTEX 572), Botryococcus sp. GE 24, Scenedesmus quadricauda, Phormidium sp. GE 2, and Spirulina maxima (UTEX 2342), C. vulgaris was selected for its fast growth and abilities to remove nitrogen and phosphorus and to produce algal biomass from swine wastewater. C. vulgaris grew well at 35$^{\circ}C$, and the optimum initial pH for growth was 8.0. In the effect of light intensity, the growth of C. vulgaris was limited under a light intensity of less than 40 ${\mu}$E/$m^2$/s. The secondary treated swine wastewater contained 58.7 mg/l of total nitrogen and 14.7 mg/l of total phosphorus, and was diluted to 75, 50, and 25% with groundwater to be treated. Nitrogen and phosphorus were removed by C. vulgaris in all diluted swine wastewaters among which the most effective removal was in 75% swine wastewater (swine wastewater:groundwater=3:1). There was a tendency of linear increase in nitrogen and phosphorus removal time with increasing concentration of swine wastewater. Under the optimized culture condition, total nitrogen and total phosphorus were effectively removed to 95.3% and 96.0%, respectively, in 25% swine wastewater after 4-day incubation.

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