• Title/Summary/Keyword: Centrifugation temperature

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Kinetic Properties of Extracted Lactate Dehydrogenase and Creatine Kinase from Mouse Embryonic Stem Cell- and Neonatal-derived Cardiomyocytes

  • Zonouzi, Roseata;Ashtiani, Saeid Kazemi;Hosseinkhani, Saman;Baharvand, Hossein
    • BMB Reports
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    • v.39 no.4
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    • pp.426-431
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    • 2006
  • Embryonic stem cells (ESCs), representing a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics, are capable of spontaneous differentiation into cardiomyocytes. The present study sought to define the kinetic characterization of lactate dehydrogenase (LDH) and creatine kinase (CK) of ESC- and neonatal-derived cardiomyocytes. Spontaneously differentiated cardiomyocytes from embryoid bodies (EBs) derived from mouse ESC line (Royan B1) and neonatal cardiomyocytes were dispersed in a buffer solution. Enzymes were extracted by sonication and centrifugation for kinetic evaluation of LDH and CK with spectrophotometric methods. While a comparison between the kinetic properties of the LDH and CK of both groups revealed not only different Michaelis constants and optimum temperatures for LDH but also different Michaelis constants and optimum pH for CK, the pH profile of LDH and optimum temperature of CK were similar. In defining some kinetic properties of cardiac metabolic enzymes of ESC-derived cardiomyocytes, our results are expected to further facilitate the use of ESCs as an experimental model.

Alcohol Dehydrogenase Active on Furfuryl Alcohol from Pseudomonas sp. (Part 1) Purification and Properties of Alcohol Dehydrogenase (Pseudomonas 속균이 생산하는 Alcohol Dehydrogenase에 관한 연구 (제1보) Alcohol Dehydrogenase 정제와 일반적성질)

  • ;Hirosake Okadar
    • Microbiology and Biotechnology Letters
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    • v.8 no.1
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    • pp.27-32
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    • 1980
  • We isolated a strain of Pseudomonas sp. from soil to utilize furfuryl alcohol as a carton source by enrichment culture. Alcohol dehydrogenase from this bacteria was purified 700-fold by Sephadex G-200 and affinity column chromatography to be homogeneous by electrophoresis and analytical centrifugation. This enzyme had a molecular weight of 120,000 and was composed of four subunits consisting of 266 amino acid residues. The optimal pH of the enzyme was pH 8.5 to 9, and the optimal temperature was, 45$^{\circ}C$. This enzyme was stable at 55$^{\circ}C$, but lost 80% of its activity in 10min at 6$0^{\circ}C$.

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Biotransformation of Amides to Acids Using a Co-Cross-Linked Enzyme Aggregate of Rhodococcus erythropolis Amidase

  • Park, Hyun-Joo;Uhm, Ki-Nam;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.325-331
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    • 2010
  • Rhodococcus erythropolis amidase was expressed in Escherichia coli cells. The crude amidase in the cell-free extract was immobilized using the cross-linked enzyme aggregate (CLEA) method. The crude amidase was mixed with bovine serum albumin and then precipitated with ammonium sulfate. The resultant precipitant was subsequently cross-linked with glutaraldehyde. Scanning electron microscopy revealed that this co-CLEA had a ball-like shape with a diameter of approximately $1\;{\mu}m$. This co-CLEA evidenced hydrolytic activity toward a variety of amide substrates. The amidase co-CLEA evidenced an optimum temperature of $60^{\circ}C$ and an optimum pH of 8.0, results that were similar to those of the soluble amidase. The reaction stability of the co-CLEA was increased. That is, it was stable up to $50^{\circ}C$ and in a pH range of 5.0-12.0. Additionally, the co-CLEA could be recovered by centrifugation, and retained 96% activity after 3 repeated cycles. This amidase co-CLEA may prove useful as a substitute for soluble amidase as a biocatalyst in the pharmaceutical and chemical industries.

Meiobenthic Community Structure on the Northeast Coastal Area of Jeju Island, Korea (제주 북동부 연안 조하대에 서식하는 중형저서동물 군집 변동 특성)

  • Kang, Teawook;Kim, Dongsung
    • Ocean and Polar Research
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    • v.42 no.1
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    • pp.1-13
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    • 2020
  • We analyzed the characteristics of meiofaunal communities in the northern part of Jeju Island and the environmental factors affecting the meiofaunal communities. For the analysis of the communities, subtidal sediments containing meiofauna were collected from 6 sites (3 shallow sites and 3 deeper sites) on the northern coast of Jeju Island in April, June, August, and October 2017. The sediment samples were collected by SCUBA using a syringe. After isolating and counting meiofauna from the sediment using a series of sieves and centrifugation, the density of the meiofauna was expressed as individuals/10 ㎠. Density of meiofauna was lower in August compared to April and June, and the density determined in October was higher than August. During April and June, the meiofaunal communities at the shallow sites (i.e., ST 1, 2, and 3) were significantly different from the meiofaunal communities at deeper sites (ST 4, 5, and 6, p < 0.01). However, such spatial variation in the meiofaunal communities was not obvious in August and October. The most dominant meiofaunal taxa were harpacticoids, followed by nematodes and nauplius larvae. These three taxa accounted for 90% of the total meiofauna density. Abundance of harpacticoids and nematodes showed a significantly negative correlation with the water depth. BIO-ENV analysis indicated that the total organic nitrogen (TON) and the water temperature exerted a significant influence over the meiofaunal communities at the sampling sites.

STUDIES ON THE VIRUSES OF RADISH MOSAIC

  • KIM, Woon-Soo
    • Journal of Plant Biology
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    • v.6 no.2
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    • pp.9-21
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    • 1963
  • A mosaic diseased radish collected from the suburb of Seoul, in November, 1961 was used for studing the host range, physical properteis, purificaitiion, insect transmission, and electron microscopy. A Japanese strain of radish mosaic(RPV) was also used with Korean strain of radish mosaic (KRMV) for a comparative study. The two viruses, KRMA and RPV, were identified by the difference in host range, insect transmission and electron microscopy. The KRMA was severely infective on tobacco and Nicotiana glutinosa, while on Gomphrena globsa was immune to the virus. RPV produces necrotic local lesions on Gomphrena globosa but did not infect tobacco and N. glutinosa. Among varieties of radish, Seoul, Akamaroo, Akanagea, Koong-Joong showed more severe symptoms than Simoo, Minong, Paek-soo, which appeared to be fainly resistant. In a number of tests, it was found that the virus KRMA retained its infectivity until to a dilution of 1:2,000, heating at $58^{\circ}$ for 10 minutes, adn aging in vitro for 7 days at room temperature. The RPV was not inactivated until it was diluted to 1:2,000, heated to $56^{\circ}$, and aged for 6 days. The KRMV was readily transmitted by the aphid(Myzus persicae Sulz). The virus RPV was not transmitted by the aphid in a number of tests. Partialy purified viruses using ammonium acetate buffer, salting-out by ammonium sulfate and centrifugation of high and low speed were highly infective. Electron micrographs showed that the KRMV paticles are of spherical particles whereas the RPV particles are rod-shaped.

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STUDIES ON THE VIRUSES OF RADISH MOSAIC

  • Kim, Woon Soo
    • Journal of Plant Biology
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    • v.6 no.2
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    • pp.7-7
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    • 1963
  • A mosaic diseased radish collected from the suburb of Seoul, in November, 1961 was used for studing the host range, physical properteis, purificaitiion, insect transmission, and electron microscopy. A Japanese strain of radish mosaic(RPV) was also used with Korean strain of radish mosaic (KRMV) for a comparative study. The two viruses, KRMA and RPV, were identified by the difference in host range, insect transmission and electron microscopy. The KRMA was severely infective on tobacco and Nicotiana glutinosa, while on Gomphrena globsa was immune to the virus. RPV produces necrotic local lesions on Gomphrena globosa but did not infect tobacco and N. glutinosa. Among varieties of radish, Seoul, Akamaroo, Akanagea, Koong-Joong showed more severe symptoms than Simoo, Minong, Paek-soo, which appeared to be fainly resistant. In a number of tests, it was found that the virus KRMA retained its infectivity until to a dilution of 1:2,000, heating at $58^{\circ}$ for 10 minutes, adn aging in vitro for 7 days at room temperature. The RPV was not inactivated until it was diluted to 1:2,000, heated to $56^{\circ}$, and aged for 6 days. The KRMV was readily transmitted by the aphid(Myzus persicae Sulz). The virus RPV was not transmitted by the aphid in a number of tests. Partialy purified viruses using ammonium acetate buffer, salting-out by ammonium sulfate and centrifugation of high and low speed were highly infective. Electron micrographs showed that the KRMV paticles are of spherical particles whereas the RPV particles are rod-shaped.

Fabrication of Transparent Heat-element using Single- Walled Carbon Nanotubes

  • Jeong, Hyeok;Vanquy, Nguyen;Lee, Han-Min;Kim, Dong-Hyeon;Kim, Do-Jin
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.05a
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    • pp.31.2-31.2
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    • 2009
  • In this research, single walled carbon nano-tube film was manufactured with spray coating method on glass for application as transparent heat element. SWNTs solution to be used for spraying is obtained by dispersion of 0.01 wt% purified SWNTs in dimethylformamide (DMF) solution through ultrasonification and centrifugation. The transmittance and sheet resistance of SWNTs film were determined by the number of spray injection. Manufactured SWNTs film will have sheet resistance range of $200\;\Omega/\square-900\;\Omega/\square$ at transmittance range of 70-90 %. Heat generation characteristic of SWNTs film was measured by applying constant DC voltage of 15V. The result confirmed that SWNTs film with sheet resistance of $200\;\Omega/\square$ reaches surface temperature of $80^{\circ}C$ within several seconds. In addition, PET coating film was coated on top of the SWNTs film by using laminator in order to solve weak adhesive property of the spray coated SWNTs film on the substrate as well as to maintain its electrical and optical properties.

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Studies on the Light-Induced Mitochondrial ATP Synthase in Pleurotus ostreotus (느타리버섯 중의 광감응성 Mitochondrial ATP Synthase 에 관한 연구)

  • Lee, Kap-Duk;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.17 no.4
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    • pp.177-183
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    • 1989
  • Mitochondria in Pleurotus ostreatus was purified by stepped sucrose density gradient centrifugation. The mitochondrial ATP synthase was investigated by various waveof the illumination at dark room for 30 min. The mitochondrial ATP synthase activity was stimulated 2.3 fold by 480 nm illumination compared with the broad wavelength group. The mitochondrial ATP synthase activity according to various times of illumination was stimulated 4.2 fold for 15 min at 480 nm compared with the broad wavelength group. The optimum pH and optimum temperature of the mitochondrial ATP synthase were 7.5 and $56^{\circ}C$, respectively. The activity of this enzyme was stimulated by 0.5 mmol $Fe^{2+}$, 1.0 mmol $Fe^{3+}$ and 5.0 mmol $k^+$ ion, but inhibited by 0.1 mmol $Na^+$ ion.

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Electron Microscopical Property of Transglutaminase Added Milk (트랜스글루타미나제를 첨가한 우유의 전자현미경적 특성)

  • 문정한;홍윤호
    • Food Science of Animal Resources
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    • v.23 no.4
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    • pp.350-355
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    • 2003
  • Raw skim milk and colloidal calcium phosphate-free skim milk were treated with microbial transglutaminase (TGase), ultracentrifuged at varying rates and were observed to contain textural properties using a scanning electron microscope (SEM). Skim milk showed irregular signs of conformation at lower centrifugal rate, and associated regular (10,000 ${\times}$g) and thin with broad holes (20,000 ${\times}$g). The associated texture became thick and irregular (40,000 ${\times}$g), and fine particles were regularly associated (100,000 ${\times}$g). When skim milk was incubated for 1 hr with TGase, casein micelles aggregated and broadened as centrifugation rate increased. When skim milk was incubated for 8 hrs with TGase, casein micelles associated to large widened aggregates, and were associated regularly which then became irregular (100,000 ${\times}$g). When colloidal calcium phosphate-free skim milk incubated for 1 hr with TGase showed no sediment, the milk incubated for 8 hrs with TGase associated together, yielding broadened and regular layers as the centrifugation rate increased. It is assumed that such phenomena could be caused by protein crosslinking reaction with TGase and conformational change of casein molecules, as well as dependencies on reaction time, temperature and ultracentrifugation rate.

Effect of Heat-Treat Methods on the Soluble Calcium Levels in the Commercial Milk Products

  • Yoo, Sung-Ho;Kang, Seung-Bum;Park, Jin-Ho;Lee, Kyung-Sang;Kim, Jin-Man;Yoon, Sung-Sik
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.369-376
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    • 2013
  • Milk is well known to be rich in some nutrients such as protein, calcium, phosphorus, and vitamins. In particular, absorption and bioavailability of calcium receive lots of attention because calcium is very little absorbed until it is changed to the ionized form in the intestine. In this study, concentration of the soluble calcium was determined in the commercial bovine milk products, which were processed by different heat-treatment methods for pasteurization. As for general constituents, lactose, fat, protein, and mineral were almost same in the liquid milk products by different processors. Ultrafiltration of the skimmed milk caused little change in the permeate as for lactose content but both fat and protein decreased. pH values ranges from 6.57-6.62 at room temperature and slightly increase after centrifugation, 10,000 g, 10 min. Rennet-coagulation activity was the lowest in the ultra high temperature (UHT-)milk compared to the low temperature long time (LTLT-) and high temperature short time (HTST-)milk products. Each bovine milk products contains 1056.5-1111.3 mg/kg of Ca. The content of sulfhydryl group was the lowest in raw milk compared to the commercial products tested. For the skimmed milks after ultrafiltration with a membrane (Mw cut-off, 3 Kd), soluble Ca in the raw milk was highest at 450.2 mg/kg, followed by LTLT-milk 336.4-345.1 mg/kg, HTST-milk 305.5-313.3 mg/kg, UHT-milk 370.3-380.2 mg/kg in the decreasing order. After secondary ultrafiltration with a membrane (Mw cut-off, 1 kD), total calcium in raw milk had a highest of 444.2 mg/kg, and those in the market milk products. As follow: UHT-milk, 371.3 to 378.2 mg/kg; LTLT-milk, 333.3 to 342.2 mg/kg; HTST-milk 301.9 to 311.2 mg/kg in a decreasing order.