• Title/Summary/Keyword: 유지 정제

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Preparation of an Immobilized Enzyme for Enhancing Thermostability of the Crude Proteinase from Fish Intestine (어류 내장 유래 단백질 분해효소로부터 열안정성 개선을 위한 고정화 효소의 제조)

  • 전유진;박표잠;변희국;송병권;김원석;김세권
    • Journal of Life Science
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    • v.8 no.6
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    • pp.627-637
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    • 1998
  • In order to utilize tuna pyloric caeca among fish intestines wasted when treated raw fish in fish processing manufactory, a crude enzyme with high proteolytic activity was extracted and its optimum condition were investigated. An immobilized enzymes also were prepared by adsorption method to enhance thermostability of the crude proteinase. The yield of the crude proteinase was approximately 2.7% on dry basis. The proteolytic activity for casein was 0.54 U/mg protein, for BTEE 1.10 U/mg protein, and for BAEE 2.69 U/mg protein. It was almost similar to that of the commercial trypsin purified. Optimum hydrolysis activity of the crude proteinase was about 80%, as the degree of hydrolysis for casein, at pH 10.0 and 45$^{\circ}C$ for 12 hrs. Also, when the crude proteinase was immobilized on DEAE-Cellulose and chitin, the residual activities remained after 7 days of pre-incubation time were maintained about 90% or more and their thermostabilities were enhanced by about 50%, compared with the native enzyme.

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Purification and Characterization of Carotenoprotein from the Muscle of Ascidian, Halocynthia roretzi (우렁쉥이 육으로부터 Carotenoprotein의 분리.정제 및 특성)

  • Kang, Ok-Ju;Byun, Hee-Guk;Jeon, You-Jin;Kim, Jong-Bae;Kim, Se-Kwon
    • Applied Biological Chemistry
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    • v.41 no.1
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    • pp.31-41
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    • 1998
  • A carotenoprotein from the muscle of Ascidian (Halocynthia roretzi) was purified by ion exchange chromatography and gel filtration chromatography, and analyzed molecular weight, stability of pH and heat, effect of denaturing agents, amino acids and fatty acids composition. The purified carotenoprotein had absorption maxima of 463 nm and 439 nm. The carotenoprotein had an approxmimate molecular weight of 64.4 kDa in polyacrylamide gel electrophoresis. The amino acid compositions of carotenoprotein were mainly Gly (15.39%), Asn (11.31%), Gln (10.62%) and Ser (13.35%). The major fatty acids composition of carotenoprotein were $C_{16:1(n-7)}\;(15.4%)$, $C_{22:1(n-9)}\;(14.5%)$ and $C_{20:1(n-11)}\;(11.4%)$. The monounsaturated fatty acids (45.2%) contained abundant content compared to other saturated (38.1%) and polyunsaturated (11.7%) fatty acids.

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Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Condition Optimization for Overexpression of the Aklavinone 11-Hydroxylase Gene from Streptomyces peucetius subsp. caesius ATCC 27952 in Escherichia coli. (Streptomyces peucetius subsp. caesius ATCC 27952 유래 Aklavinone 11-Hydroxylase 유전자의 대장균에서의 대량발현과 최적화)

  • 민우근;홍영수;최용경;이정준;홍순광
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.15-22
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    • 1998
  • The dnrF gene, responsible for conversion of aklavinone to $\varepsilon$-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin gene cluster of Streptomyces peucetius subsp. caesius ATCC 27952, close to drrAB, one of the anthracycline resistance genes. To characterize the enzymatic properties of the aklavinone 11-hydroxylase, the dnrF gene was overexpressed in Escherchia coli. The pET-22(+) plasmid which has the T7 promoter under the control of lacUV5 gene was used for the overexpression of the dnrF gene, and the recombinant plasmid pET213 that contains the dnrF gene linked to the T7 promoter of pET-22b(+) was introduced into the E. coli BL2l. When the expression of the dnrF gene was induced by IPTG at the final concentration of 1 mM, the induced protein could be detected in SDS-PAGE only in insoluble precipitate. The insoluble protein was electroeluted from the gel and used for the preparation of antiserum in mice. Various culture conditions were tested to maximize the expression of the aklavinone 11-hydroxylase in soluble form. The enzymatic activity was checked by the bioconversion experiment, and the protein was confirmed by the SDS-PAGE and the Western blot analysis. From the analysis of the data, it was concluded that the culture induced with IPTG at the final concentration of 0.02 mM at 37$^{\circ}C$ yielded the best productivity of active form of enzyme.

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Immuno-stimulating Activities of Mannose-rich Polysaccharides Isolated from Korean Black Raspberry Wine (복분자주에서 분리한 Mannan 다당의 면역증진 활성)

  • Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.343-349
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    • 2012
  • Polysaccharides isolated from Korean black raspberry wine were examined for their chemical properties and immuno-modulating activities. The molecular mass of RB-1b-I, the main polysaccharide in black raspberry wine, was estimated as 180 kDa and it contained a significant proportion of mannose (76.8%) and 4 different minor component sugars such as galactose (15.8%), arabinose (3.8%), glucose (2.6%) and rhamnose (1.2%). This indicated that RB-1b-I was mainly present as a mannan, which had originated from the cell walls of fermenting yeasts. On the other hand, RB-1b-I induced high levels of macrophage activation as well as mitogenicity regarding murine splenocytes in vitro. The intravenous administration of RB-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. RB-1b-I also showed potent anti-complementary activity in a dose-dependent manner via both alternative and classical pathways. Results indicated that Korean black raspberry wine contains peculiar polysaccharides which provide beneficial immuno-stimulating activities for human health.

Reason and Prevention of Color Reversion of Corn Oil in Summer (옥수수기름의 하절기 변색현상 원인규명 및 방지대책)

  • Koo Bon-Soon
    • Food Science and Preservation
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    • v.11 no.4
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    • pp.485-490
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    • 2004
  • Crude corn oil (CCO) was obtained through the expression-extraction process from corn germ. The CCOs of final process at $90^{\circ}C,\;70^{\circ}C\;and\;50^{\circ}C$ were stored in outdoor storage tanks. From the samples, refined CCO (RCCO) were prepared with $0\%,\;10\%\;and\;20\%$ excessive of phosphoric acid, caustic solution and acidic clay were used in degumming-alkali refining-bleaching process. RCCOs were stored at room temperature in dark places. The color change was not effected by the amount of phosphoric acid, caustic solution and acidic clay, but temperature of process affects the color change. Finally, the prevention for color reversion of RCCO could be obtained by lowering the temperature of final process and optimal temperature of RCCO in summer was found about less than $50^{\circ}C$.

The Oxidation Stability and Flavor Acceptability of Oil from Roasted Soybean (볶은 대두로부터 착유한 대두유의 산화 안정성 및 향기 기호도)

  • Im, Moo-Hyeog;Choi, Jong-Dong;Choi, Kwang-Soo
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.425-430
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    • 1995
  • The effects of oxidation stability and flavor acceptability of soybean oil by roasting and refining treatment were investigated in this study. The peroxide values of crude oil from unroasted soybean(COUS) were more increased than those of crude oil from unroasted soybean(CORS) during storage under fluorescence light at $45^{\circ}C$. Induction time by the Rancimat Method was 44.9 and above 88.7 hours on COUS and CORS respectively. The order of oxidation stability of oil was crude>degummed>alkali-refining>deodorized>bleached oil in terms of peroxide value, while that of oxidation stability of oil was crude>degummed>deodorized>alkali-refining>bleached oil in regard to induction time by the Rancimat Method. The correlation coefficients between induction time and two charateristics(absorbance of browning reaction products and phosphorous contents) were highly significant, while that between induction time and tocopherol contents was not high during refining stages. The scores for the sensory evaluation of flavor on sesame oil and CORS was 9.269 and 8.269 respectively, but it was not significant between two oils.

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Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus (Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명)

  • Cha, Hyun A;Chung, Dawn;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.393-401
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    • 2014
  • A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

Purification and Characterization of an Invertase Produced with Saccharomyces cerevisiae JS59 Isolated from Home-made Wine (포도주에서 분리한 Saccharomyces cerevisiae JS59가 생성하는 Invertase의 정제 및 특성)

  • Yoo, Ji-Soo;Paik, Hyun-Dong;Kim, Soo-Young;Lee, Si-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1321-1327
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    • 2011
  • The microorganism producing an invertase (E.C. 3.2.1.26) was isolated from wine and tentatively identified as Saccharomyces cerevisiae by cellular fatty acid analysis. The invertase was purified to homogeneity by ammonium sulfate precipitant, dialysis, ion-exchange chromatography on DEAE-Sephadex A-50, and gel chromatography on Sephadex G-200 from the culture supernatant of Saccharomyces cerevisiae JS59. The specific activity and the purification fold of the purified invertase were 7620.9 unit/mg protein and 13.9, respectively. The molecular weight of the purified invertase was estimated to be 38.5 kDa by SDS-PAGE. The optimum pH and temperature for the invertase activity were pH 5 and $55^{\circ}C$, respectively. The invertase activity was relatively stable at pH 4~6 and temperature $55^{\circ}C$. The activity of invertase was inhibited by $Ag^{2+}$ and $Hg^{2+}$, but on the contrary, activated by $Co^{2+}$ and $Mn^{2+}$. Michaelis constant ($K_m$) for invertase reaction in sucrose solution was 11.5 mM. TLC analysis of the products produced in sucrose solution during invertase reaction showed the progressive presence of glucose and fructose in accordance with sucrose hydrolysis.

Study of UV Degradation of Lacquer and Natural Adhesives Using Lacquer Mixed with Animal Glue (옻과 옻에 아교를 배합한 천연접착제의 자외선에 의한 노화 특성 연구)

  • Ahn, Sunah;Kim, Eun Kyung;Jang, Sungyoon
    • Journal of Conservation Science
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    • v.32 no.4
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    • pp.501-510
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    • 2016
  • In this study, we investigated the degradation of adhesives when exposed to ultraviolet light irradiation using samples of lacquer (L), treated lacquer (TL), lacquer mixed with glue (LG), and urushiol mixed with glue (UG). Four types of film specimens were collected under the ultraviolet exposure time, and gloss test, tensile shear strength test, scanning electron microscope analysis, and infrared spectroscopic analysis were conducted for the specimens. LG and UG showed lowering rate of gloss is somewhat later than L. Also, it was observed that with increasing exposure time to ultraviolet irradiation, the surface of L began to show spherical pits and cracks when the polysaccharide layers started to be exposed, whereas the surfaces of LG and UG remained smooth. The Infrared spectra of L and TL showed that the intensity of the overall peak decreased with increasing ultraviolet irradiation time. There was no change in the peak intensity of LG, but for UG, the peaks at $3013cm^{-1}$, $1593cm^{-1}$ and so on disappeared and the overall intensity declined. The tensile shear strength of LG and UG was maintained or increased as compared to the initial test, whereas the tensile shear strength of L decreased sharply after 600 h. LG and UG exhibited fewer changes as a result of high temperature and humidity conditions, and they retained their strength under UV exposure. These results indicate that LG and UG are more durable than L when subjected to environmental change.