• Title/Summary/Keyword: sucrose density gradient

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Development of Curdlan Separation Process with Density Gradient Centrfugation (Density Gradient를 이용한 식품소재를 커들란의 분리공정개발)

  • 김봉영;이중헌
    • KSBB Journal
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    • v.16 no.5
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    • pp.523-525
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    • 2001
  • Curdlan is one biopolymer composed of ${\beta}$1,3-glucan and dissolved in a alkali solution but formed salt under neutral or acid condition. It was produced by Agrobactrium species and the separation process is necessary to make pure curdlan from the culture broth. The pH swing separation method was as feasible separation process using solubility changes with the pH difference. however, this method requires a lot of acid and alkali solution also produces a lot of waste. Therefore, an efficient process which could save energy and minimize toxic waste was developed. A density gradient separation process was developed in this research. High density sucrose solution was used as a separation agent. Curdlan was separated from the culture broth when the density of the sucrose solution was 1.15 g/L. Since the curdlan was produced on the surface of cell wall. the pre-treatment of culture broth was necessary. Curdlan recovery yield was increased up to 83% with the homogenization of the culture broth and further increased up to 87% with the treatment of alkai-acid solution.

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Separation of soil Organic Debris using Sucrose-ZnCl2 Density Gradient Centrifugation

  • Jung, Seok-Ho;Chung, Doug-Young;Han, Gwang-Hyun
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.1
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    • pp.30-36
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    • 2012
  • The active fraction of soil organic matter, which includes organic debris and light organic fraction, plays a major role in nutrient cycling. In addition, particulate organic matter is a valuable index of labile soil organic matter and can reflect differences in various soil behaviors. Since soil organic matter bound to soil mineral particles has its density lower than soil minerals, we partitioned soil organic matter into debris ($<1.5g\;cm^{-3}$), light fraction ($1.5-2.0g\;cm^{-3}$), and heavy fraction ($>2.0g\;cm^{-3}$), based on high density $ZnCl_{2-}$ sucrose solutions. Generally, partitioned organic bands were clearly separated, demonstrating that the $ZnCl_{2-}$ sucrose solutions are useful for such a density gradient centrifugation. The available gradient ranges from 1.2 to $2.0g\;cm^{-3}$. Although there was not a statistically meaningful difference in organic debris and organomineral fractions among the examined soils, there was a general trend that a higher content of organic debris resulted in a higher proportion of light organomineral fraction. In addition, high clay content was associated with increased fraction of light organomineals. Partitioning of soil organic carbon revealed that carbon content is reduced in the heavy fraction than in the light fraction, reflecting that the light fraction contains more fresh and abundant carbon than the passive resistant fraction. It was also found that carbon contents in the overall organic matter, debris, light fraction, and heavy fractions may differ considerably in response to different farming practices.

Relatedness Between Different Morphologies of L. interrogans Isolated from a Patient with Epidemic Pulmonary Hemorrhagic Fever (유행성 폐출혈열 환자에서 분리된 병원체에서 관찰된 다양한 형태들간의 연관성)

  • Lee, Tae-Yoon;Kim, Kyung-Won;Kim, Gye-Sung
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.219-224
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    • 1987
  • A strain of Leptospira interrogans(UM-19) was isolated in 1984 from a patient with epidemic pulmonary hemorrhagic fever. The bacteria has been characterized morphologically by a polymorphism showing spiral forms, short and long rods in culture. In order to study the biological relatedness among these forms of the bacteria, the short rods were separated from others by means of continuous sucrose density gradient centrifugation, and cultured at various temperatures such as $10^{\circ}C$, $15^{\circ}C$, $30^{\circ}C$ and $37^{\circ}C$. It was revealed that cultivation or subcultivation of the short rods has resulted in morphological changes observed by darkfield examination, silver stain, and scanning electron microscope showing spiral forms as well as short and long rods.

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Histidylleucine Hydrolyzing Enzyme : A Dipeptidase From Hog Lung (Dipeptidase로서의 Histidylleucine 가수분해 효소)

  • Hyun-Jae Lee;Jung-Keun Park
    • Journal of the Korean Chemical Society
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    • v.19 no.5
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    • pp.375-380
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    • 1975
  • An enzyme capable of hydrolyzing histidylleucine was purified 50 fold from hog lung. The final preparation hydrolyzed $1.6{\mu}moles$ of histidylleucime per minute per mg of protein. The $K_m$ of the enzyme for the enzyme was found to be $2{\times}10^{-4}M$. The enzyme was required a number of free dipeptides for the substrate specificity, and was inhibited by EDTA and 1,10-phenan-throline. The molecular weight of the enzyme was estimated to be 80,000 daltons from sucrose density gradient sedimentation analysis. The corrected $s_{20,w}$ value was 5.3 S.

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Isolation and Identification of Cyanophage from Eutrophic Water (부영양화 수역에서의 Cyanophage 의 분리와 동정)

  • Kim, Min;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.524-527
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    • 1992
  • Synechococcus sp. cyanophage was isolated from Baekwoon reservoir located in KyonggiDo. The cyanophage was purified by employing ultrafiltration. differential centrifugation. and sucrose density gradient centrifugation. Electron microscopic observation indicated that the sizes of its isometric head and contractile tail are 89 nm and] II nm. respectively. which means that the isolated cyanophage is included in the group. Myoviridae. The cyanophage maintained the stability of more than 50 percent from $20^{\circ}C$ to $40^{\circ}C$ and from pH 5 to 8. and had the maximal infectivity at $30^{\circ}C$ and pH 9 implying its ecological significance.

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Localization of Lipoxygenase in Germinating Soybeans (발아중인 대두콩에서의 Lipoxygenase의 국재(局在))

  • Song, Young-Sun
    • Korean Journal of Food Science and Technology
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    • v.19 no.5
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    • pp.441-445
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    • 1987
  • The subcellular distribution of lipoxygenase in germinating soybean seeds (Glycine max[L.] AmSoy) was investigated by using differential centrifugation and sucrose density gradient fractionation. Most of lipoxygenase -1 and -2/3 activities was present in the supernatant fraction after differential centrifugation of homogenates prepared from three-day-old seedlings; only 1.5% of lipoxygenase activity remained in particulate fractions. The results of a sucrose density gradient fractionation (three-day-old) showed that the lipoxygenase activity coincided with acid phosphatase at the densities of 1.19, 1.23, $1.25g/cm^3$, even though most of lipoxygenase and acid phosphatase activities appeared in supernatant fractions. There was no indication that mitochondria contained any lipoxygenase activity, and it does not appear that glyoxysomes and ER contained any lipoxygenase activity either.

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Phycobilisome composition in Chondrus crispus (Gigartinales, Rhodophyta) from a wild type strain and its vegetatively derived green mutant

  • Cornish, M. Lynn;O' Leary, Stephen J.B.;Garbary, David J.
    • ALGAE
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    • v.28 no.1
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    • pp.121-129
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    • 2013
  • Intact phycobilisomes from a wild-type red Chondrus crispus and its vegetatively derived green mutant were isolated by centrifugation through a discontinuous sucrose density gradient. Pigment composition was subsequently characterized by spectrophotometry. Vegetative thalli of the two strains grown together for six months in the laboratory resulted in different pigment profiles. Two pigmented phycobilisome bands appeared in the sucrose gradient of the wild-type alga, a purple coloured one, and a pink one, whereas only a single blue band appeared in the gradient of the green mutant. Spectrophotometric and fluorescence analyses identified the phycobiliprotein composition of the purple band as the typical phycoerythrin-phycocyanin-allophycocyanin complement in the wild-type, but there was no detectable phycoerythrin present in the blue band of the green mutant. Sodium dodecyl sulphate, preparative polyacrylamide gel electrophoresis analysis confirmed the presence of allophycocyanin subunits in all extracts, but firm evidence of an R-phycoerythrin linker polypeptide in the blue band was missing. These results highlight the ability of C. crispus to adapt to a phycoerythrin deficiency by adjusting light harvesting pigment ratios.

Comparison of Several Procedures for the Preparation of Synaptosomal Plasma Membrane Vesicles

  • Yun, Il;Kim, Young-Shin;Yu, Seong-Ho;Chung, In-Kyo;Baik, Seung-Wan;Cho, Goon-Jae;Chung, Yong-Za;Kim, Seok-Hwan;Kang, Jung-Sook;Kim, In-Se
    • Archives of Pharmacal Research
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    • v.13 no.4
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    • pp.325-329
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    • 1990
  • Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex by several well-known procedures, and the best procedures was selected by enzymatic and morphological standards. The SPMV isolated by the modified procedure of Smith and Loh showed the highest purity. The specific activities of Na, K-ATPase, acetylcholinesterase and 5'-nucleotidase were about 5, 6-fold, 2, 5-fold and 3, 3-fold, respectively, enriched in the plasma membrane fraction as compared to those of the crude homogenate. Electron microscpic examination also showed that the membranes were in vesicular form.

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Studies of Density-Fractionated Human Ervthrocvte Membranes (농도분배에 따라 분리한 사람 적혈구 막에 관한 연구)

  • 정종문
    • The Korean Journal of Zoology
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    • v.37 no.4
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    • pp.597-604
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    • 1994
  • Membranes obtained from the normal human RBC population were separated by continuous sucrose density gradient centrifugation and the density-fractionated membranes were then examined for changes in molecular markers. This study focuses on changes of (i) the membrane protein profile, (ii) differences in membrane-associsted enzyme activities, and (iii) the amount of autologous IgG bound. The following observations were made: (i) ratios for band 4. la over the sum of bands (4. la + 4.Ib) ranged from 0.58 to 0.79 for membranes of lowest density; (ii) significant changes in bound glyceraldehyde-3-phosphate dehydrogenase and acetvlcholinesterase activities were found; (iii) the amounts of autolosous IgG's attached to the red blood cells was highest in the membrane fraction of lowest density.

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