• Title/Summary/Keyword: 단일 효소 반응

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Enzymatic Synthesis of New Oligosaccharides Using Glucansucrases. (Glucansucrases를 이용한 새로운 올리고당의 합성)

  • ;;;;;John F. Robyt
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.179-186
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    • 1998
  • Dextransucrase hyper-producing Leuconostoc mesenteroides B-512FMCM and dextransucrase constitutive mutants B-742CB and B-1355C catalyzed the transfer of glucose from sucrose to other carbohydrates which were present or were added to the reaction digests. When the acceptor was a maltose, gentiobiose, lactose or raffinose, there was produced a series of oligosaccharide acceptor products or single product based on the kinds of enzymes and reaction conditions. To obtain the quantitative information about the yield and the distribution of acceptor products and dextran two experimental parameters were studied: a) the ratio of acceptor to sucrose and b) the amount of enzyme at constant carbohydrate concentration (100 mM). As the amount of enzyme increased, the synthesis of acceptor products (of maltose or gentiobiose) increased, and the formation of dextran decreased. As the ratio of acceptor to sucrose increased, the amount of dextran and the number of acceptor-products decreased and the amount of acceptor-products increased. When maltose or gentiobiose was an acceptor, the glucose from sucrose was transferred to the C-6 hydroxyl group of the nonreducing-end glucose residue of accepters to give a homologous series of isomaltosyl dextrins. In case of lactose or raffinose, there was produced only one acceptor product from B-512FMCM dextransucrase reaction. In the lactose acceptor reaction, the glucose from sucrose was transferred to the C-2 hydroxyl of the reducing end glucose residue of lactose. To get a series of oligosaccharides from lactose or raffinose acceptor reaction we used B-742CB dextransucrase or B-1355C alternansucrase with 500 mM sucrose in reaction digest.

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One-Stage Polymerase Chain Reaction for the Comprehensive Detection of Type D Retrovirus Provial DNA (Type D Retrovirus 감염의 포괄적 검색을 위한 One-Stage 중합효소 연쇄반응법의 개발)

  • Jeong, Yong-Seok
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.19-27
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    • 1997
  • To develop the polymerase chain reaction (PCR) for the detection of type D simian retrovirus (SRV) infection, an oligonucleotide primer pair was designed to hybridize to the sequences within env gene of SRV subtype 1 (SRV-1). The 3' proximal env sequences annealing to the primers had been rather conserved among three different subtypes of SRV, SRV-1, SRV-2, and SRV-3 (Mason-Pfizer Monkey Virus: MPMV). The PCR using the primer pair targeting an env region successfully detected and amplified all three subtypes of SRV with excellent specificity after single round of reaction. The tests with peripheral blood mononuclear cells infected either with simian immunodeficiency virus or simian T-Iymphotropic virus type 1, major immunosuppressive viral agents together with SRV in simian, verified the specificity of the PCR by excluding any cross reactivity. Semiquantitative titration PCR, amplifying serially diluted plasmid DNA of each subtype, was performed to evaluate sensitivity limits of the reaction. Based on molecular weight of each cloned SRV genome, the PCR should be able to detect one SRV-infected cell per more than $5-7{\times}10^4$ uninfected cells after simple ethidium bromide staining of resulting products. The PCR must be very efficient screening system with its quickness, certainty, and sensitivity for SRV-infected animals used in human AIDS research model. Second round amplification of the reaction products from the first PCR, or Southern hybridization by radiolabeled probes shall render to compete its efficacy to ELISA which has been the most sensitive technique to screen SRV infection but with frequent ambiguity problem.

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Development of Biomolecular Device Using Biomolecular Film Part 1: Optical Biosensor to Detect the Ethanol Using Langmuir-Blodgett Film of Eilzyme Molecules (생체분자막을 이용한 생물분자소자의 개발 제1부 :효소분자 LB막을 이용한 에탄올 측정용 광학 바이오센서)

  • 최정우;배주연지용이원홍
    • KSBB Journal
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    • v.10 no.1
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    • pp.105-112
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    • 1995
  • The fiber-optic biosensor using enzyme-immobilized Langmuir-Blodgett film is developed fort the measurement of ethanol. The enzyme, alcohol dehydrogenase, is immobilized at the molecular level on the arachidic acid monolayer using Langmuir-Blodgett film technique. Based on the ordered multisubstrate mechanism, the immobilized enzyme kinetics is investigated. The optical sensing system is proposed, and sensor signal is proportional to ethanol concentration and is related wish the number of enzyme layers. As the number of deposited LB film layer increases up to 20 1ayers, the high ethanol concentration of 45mM can be measured without the saturation of signal. Surface pressure-area isotherm is measured for the three-different charged-lipids. Arachidic acid is the most suitable for the adsorption of alcohol dehydrogenase based on electrostatic force.

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Molecular Action of Prostaglandin to Mediate Insect Immunity and Its Application to Develop Novel Insect Control Techniques (곤충 면역반응을 중개하는 프로스타글란딘의 분자적 기작과 해충방제 응용)

  • Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.173-195
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    • 2022
  • Like vertebrates, insects synthesize various eicosanoids after the committed catalytic step of phospholipase A2 (PLA2). However, the subsequent biosynthetic steps exhibit some deviation from those of vertebrates. Due to little composition of arachidonic acid in insect phospholipids, PLA2 releases linoleic acid, which is another polyunsaturated fatty acid and relatively rich in insect phospholipids, to synthesize arachidonic acid via chain extension and desaturation. Resulting arachidonic acid is then oxygenated into a prostaglandin (PG), PGH2, by a specific peroxidase called peroxynectin, but not by cyclooxygenase. PGH2 is then isomerized to various PGs such as PGA2, PGD2, PGE2, PGI2, and a thromboxane (TXB2). All four epoxyeicosatrienoic acids such as 5,6-EET, 8,9-EET, 11,12-EET, and 14,15-EET are also synthesized from arachidonic acid by oxygenation of vertebrate types of monooxygenases. However, the other type of eicosanoids called leukotrienes are found in insect tissues but their synthetic pathway is unclear. Eicosanoids mediate various insect physiological processes such as metabolism, excretion, immunity, and reproduction. Thus, identification of novel compounds interrupting eicosanoid biosynthesis would be a novel approach to develop insecticides. This review focuses on PGs and their immune mediation.

Progression of Atherosclerosis After Angioplasty (혈관 성형술후 동맥경화의 재발생)

  • Kwon, Hyuck-Moon;Lee, Byoung-Kwon;Suh, Sang-Ho;Roh, Hyung-Woon;Kim, Gye-Young;Lee, Na-Young
    • 순환기질환의공학회:학술대회논문집
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    • 2005.12a
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    • pp.25-26
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    • 2005
  • 동맥경화의 재발생 위치는 속도와 전단응력 등과 같은 혈류역학의 인자들의 영향을 많이 받는 혈관형태를 가진 영역이다. 이러한 결과는 관상동맥에 동맥경화의 발생빈도를 조사한 결과와 일치하고 있으며, 즉 좌전 하행지, 회선지, 및 우관동맥 등의 동맥경화성 병변 발생빈도에서 좌전하행지가 가장 많은 빈도를 나타낸다. 따라서 동맥경화의 발생 및 재형성은 혈관의 동맥경화성 위험지역의 형태적 특징, 즉, 분지부의 위치, 길이, 각도의 변화 등에 따라 달라질 수 있음을 시사한다. 동일한 관상동맥이더라도 동맥경화의 발생이 용이한 형태가 있는데, 혈관의 형태학적 특성에 따란 혈류역학적 특성이 달라지고 동맥경화가 발생할 수 있는 가능성이나 진행과정이 차이가 날 수 있음을 말한다. 특히 임계치를 넘는 고전단응력은 혈관내피세포를 파괴하거나 손상을 주며, 반대로 임계치 미만의 저전단응력은 혈류의 정체시간을 길게 하여 양쪽 모두 동맥경화성 생물학적 반응을 유발 할 수 있며, 고전단응력과 저전단응력의 빈번한 맥동성 변화작용으로 혈관이 손상될 수 있는 한계범위를 넘어서게 될 때 내피세포의 방어체계를 파괴시키거나 혈관성형술후의 신내포세포 형성과정에서 생물학적 활성반응을 촉진하게 되는 환경을 제공하게 되어 동맥경화를 촉진한다고 할 수 있다. 즉 임계치 이상의 고전단응력이 나타나는 형태와 입구경계조건이 발생되면 내피세포 손상에 따른 혈전 현상의 발생가능성이 높아지며, 임계치 미만의 저전단응력이 발생되면 동맥경화성 죽상반 재형성에 영향을 미치게 한다. 결론적으로 동맥경화의 재발생의 기전은 변형된 혈관의 형태학적인 차이와 위치에 따라 서로 다른 혈류역학적 유발할 수 있는 물리적 환경을 제공하는 데에서 출발한다고 할 수 있다.$8.0{\sim}8.3$으로 알카리 쪽으로 이동하였다. 파일롯트 규모로 본 고정화 효소 충전탑(내경 30cm, 높이 85cm)에 의한 이성화당의 생산을 시도하였던바, 고정화 효소(350 IXIU/ml-R) 1리터가 30일동안에 약 293리터의 이성화당을 생산할 수 있는 것으로 나타났다.l plane에서 선수군(選手群)이 $62.7{\pm}7.36^{\circ}$로서 비선수군(非選手群)과 별(別) 차이(差異)가 없었고, horizontalplane에서는 선수군(選手群)이 $-23.5{\pm}7.2^{\circ}$로서 비선수군(非選手群)의 $-38.8{\pm}8.2^{\circ}$에 비(比)해 유의(有意)하게 높았으며 운동후(運動後) 양군(兩群) 모두 유의(有意)하게 높았다. QRS vector 길이에서 Frontal plane에서 선수군(選手群)이 $13.86{\pm}1.44\;mm$로서 비선수군(非選手群)의 $9.62{\pm}0.97\;mm$에 비(比)해 유의(有意)하게 높았으며 운동후(運動後)에도 유의(有意)하게 높았다. Horizontal plane에서도 선수군(選手群)이 $19.82{\pm}2.10\;mm$로서 비선수군(非選手群)의 $16.90{\pm}1.39\;mm$에 비(比)해 유의(有意)하게 높았고 운동후(運動後)에도 선수군(選手群)이 유의(有意)하게 높았다. 이상(以上)을 종합(綜合)해 보면 선수군(選手群)의 R파고(波高)가 비선수군(非選手群)에 비(比)해 운동후(運動後) 계속(繼續) 유의(有意)하게 높았고, $Rv_5$

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Enzymatic Characteristics of ${\alpha}-Galactosidase$ for the Removal of Flatulence Factor in Soybean (대두(大豆)의 Flatulence Factor 제거(除去)를 위한 ${\alpha}-Galactosidase$ 효소제의 특성)

  • Jung, Sang-Soo;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.18 no.6
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    • pp.450-457
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    • 1986
  • For the removal of raffinose and stachyose related to flatulence in soybean, ${\alpha}-Galactosidase$ activity of six commercial enzyme preparations was compared and their enzymatic characteristics were investigated. Among the tested enzymes, one product from Aspergillus niger was shown to be the most potent in ${\alpha}-Galactosidase$ activity. The enzyme characteristics of the selected preparation were shown to be pH 4.0-4.5 for optimum activity, pH 4-5 for optimum stability and $45^{\circ}C$ for optimum activity. Upon reaction on a synthetic substrate, $p-nitrophenyl-{\alpha}-D-galactoside$, Michaelis constant was 2.08 mM and maximum velocity was 435 micromoles of substrate/minute/g enzyme preparation. The enzyme was proved to be essential for SH group for its activity and capable of hydrolyzing raffinose, sucrose and $p-nitrophenyl-{\alpha}-D-galactoside$ almost completely. Thin-layer chromatographic analysis exhibited that the enzyme treatments of raffinose and stachyose were resulted to produce only monosaccharides in 2 hours of hydrolysis. It was, therefore, assumed that the flatulence factor in soybean foods can be easily removed by the use of enzymes showing ${\alpha}-Galactosidase$ activity.

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유리반도체

  • 박창엽
    • 전기의세계
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    • v.24 no.4
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    • pp.6-10
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    • 1975
  • 반도체와 그의 응용소자는 지난 20여년간 눈부식 발전을 이룩하였다. 이는 주로 단결정의 제작기술 진보에 의한 것으로 본다. 그러나 최근 단결정과는 전연 다른 유리질반도체가 국제회의에서도 그 우수성을 의논하기에 이르렀다. 유리질 반도체가 주목을 끌게 된겻은 1968년 Ovshinsky가 "무질서 구조에 있어서 가역적 스위칭현상"이라는 논문이 발표되고 유리질 반도체를 사용한 Ovonic 스위칭 소자의 출현에 기인된다. 유리질 반도체가 전기스위칭 작용, 기억작용을 나타낸다고 하는 Ovshinsky의 발표는 전자제치로서의 응용에 대해 찬반되는 의견이 있었지만 물성적 연구의 교량적인 역할을했다고 할 수 있다. 이런 반도체에 속하는 재료는 호칭도 여러가지로 유리질반도체, 비정질반도체 무정형반도체등으로 불리어진다. 단결정체가 각 격자간에 장거리질서를 갖는 반면 유리질 반도체는 무질서한 구조로 각 격자간에 단거리 질서를 갖는 것이 단결정과는 본질적으로 다른 점이라 본다. 유리 반도체의 종류는 첫째, 원소성 유리반도체로서 Ge, Si, Se, Te 들과 같이 단일원소로 된 겻과, 둘째 IV, V, VI족 원소로 된 공유결합 합금인 As$_{2}$Se$_{3}$-As$_{2}$Te$_{3}$ 계 Ge Si As Te계등의 칼코게나이드 유리등으로 금지대는 어느 것이나 2eV이하이다. 셋째 이론결합인 SiO $Al_{2}$O$_{3}$ Ta$_{2}$O$_{3}$Si$_{3}$N$_{4}$등의 산화물 및 질화물로 대표되는 분자성 비정질 물질로서 금지대는 2eV보다 큰 세종류로 크게 분류할 수 있다. 분류할 수 있다. 한다. 단 개개의 문제에 관한 구체적인 해석 또는 검토에 관하여는 다음 기회에 미루기로하고, 우선 여기서는 당면문제로서 대처하지 않으면 안될 자동주파수제어문제및 계통의 경제운용문제만에 한정하여, 이것을 우리나라의 현상과 관련시켜 개설하고, 이들의 자동화에 관한 기본적인 문제를 간단히 적어 보겠다. 가능하다. 제작완료된 ASIC은 기능시험을 완료했으며 실제 line-of-sight(LOS) 시스템 구현에 적용중이다. 시대를 살아 갈 회원들이다. '컨텐츠의 시대'가 개막되는 것이며, 신세기통신과 SK텔레콤은 선의의 경쟁 과 협력을 통해 이동인터넷 서비스의 컨텐츠를 개발해 나가게 될 것이다. 3배가 높았다. 효소 활성에 필수적인 물의 양에 따른 DIAION WA30의 라세미화 효율에 관하여 실험한 결과, 물의 양이 증가할수록 그 효율은 감소하였다. DIAION WA30을 라세미화 촉매로 사용하여 아이소옥탄 내에서 라세믹 나프록센 2,2,2-트리플로로에틸 씨오에스터의 효소적 DKR 반응을 수행해 보았다. 그 결과 DIAION WA30을 사용하지 않은 경우에 비해 반응 전환율과 생성물의 광학 순도는 급격히 향상되었다. 전통적 광학분할 반응의 최대 50%라는 전환율의 제한이 본 연구에서 찾은 DIAION WA30을 첨가함으로써 성공적으로 극복되었다. 또한 고체 염기촉매인 DIAION WA30의 사용은 라세미화 촉매의 회수 및 재사용이 가능하게 해준다.해준다.다. TN5 세포주를 0.2 L 규모 (1 L spinner flask)oJl에서 세포간의 응집현상 없이 부유배양에 적응,배양시킨 후 세포성장 시기에 따른 발현을 조사한 결과 1 MOI의 감염조건 하에서는 $0.6\times10^6$cell/mL의 early exponential시기의 세포밀도에서 72시간 배양하였을 대 최대 발현양을

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Purification and Properties of Cyclodextrin glycosyltransferase from Bacillus stearothermophilus KY-126 (Bacillus stearothermophilus KY-126가 생산하는 Cyclodextrin glycosyltransferase의 정제 및 특성)

  • Kang, Sang-Mo;Yoo, Si-Hyung
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.375-381
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    • 1994
  • A bacterial strain No. KY-126, which produced extracellular cyclodextrin glycosyltransferase(CGTase), was isolated from soil and identified as Bacillus stearothermophilus KY-126. The enzyme was purified by the treatments of ammonium sulfate precipitation, DEAF-Sephadex, Sephadex G-100 column chromatography. The optimal pH and temperature for the enzyme activity were pH 5.5 and $65^{\circ}C$, respectively. And the enzyme was stable at pH values from 6.0 to 11.0 at $55^{\circ}C$ for 30 min and stable up to $60^{\circ}C$ for 30 min.. The enzyme was inhibited by $HgCl_{2}$. The molecular weight of the enzyme was estimated to be 67,000 by using SDS-PAGE. The maximum conversion from starch to cyclodextrin (CD) by CGTase was 43% and obtained at 6 hr reaction and the ratio of ${\alpha}-,\;{\beta}-,\;{\gamma}-$, CD production at this time was 2.9 : 2.1 : 1.0.

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Improvement on the Quality and Functionality of Red Tanner Crab Cooking Drip Using Commercial Enzymes (효소분해의 의한 붉은 대게 자숙액의 품질 및 기능성 개선)

  • Kang, Kyung-Tae;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1022-1030
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    • 2007
  • For the improvement on the quality and functionality of red tanner crab cooking drip, the preparation of hydrolysates from red crab cooking drip using commercial enzymes (Alcalase, Flavourzyme, Neutrase and Protamex) was attempted and its taste, nutritional and functional characteristics were also investigated. According to the results of heavy metal contents and proximate composition, red tanner crab cooking drip (RTCCD) could be used as a food resource. From the results of the trichloroacetic acid soluble index (TSI), angiotensin I converting enzyme (ACE) inhibiting activity and antioxidative activity, RTCCD hydrolysates incubated with Alcalase for 2 hrs was superior to the other one-step hydrolysates. There were no differences in the ACE inhibiting activity and antioxidative activity between one-step hydrolysates, which was incubated with Alcalase for 2 hrs, and two-step hydrolysates sequentially incubated with Alcalase and other enzymes. Alcalase-treated hydrolysates was similar in proximate composition and Hunter color value, while high in free amino acid content compared with crab cooking drip. Total amino acid content of Alcalase-treated hydrolysates was 11.9 g/100 mL and the major amino acids were glutamic acid (10.2%), proline (10.1%) and glycine (10.7%).

The Esterification of Acetyltyrosine by $\alpha$-Chymotrypsin in EtOH/Water Mixture (에탄올 내에서 $\alpha$-Chymotrypsin에 의한 Acetyltyrosine의 에스테르화 반응)

  • 전유진;김세권
    • KSBB Journal
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    • v.9 no.3
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    • pp.312-318
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    • 1994
  • The esterification of Ac-Tyr-OH was carried out in one-phase system containing ethanol by ${\alpha}$-chymotrypsin. The results of the esterification reaction are as follows. Chitin-${\alpha}$-chymotrypsin complex was found to be an effective catalyst for the esterlfication of Ac-Tyr-OH in ethanol organic solvent. The optimal conditions for the esterification were chitn/${\alpha}$-chymotrypsin ratio, 20(w/w); reaction temp., $35^{\circ}C$; reaction pH, 8.0; reaction time, 24 hrs. Also, addition of chitin in water/water-miscible organic solvent was effective for the stability of the enzyme. The esterification yield, Km and Vmax under optimal conditions were 93%, 3.093mM and 1.088mM/mg/hr, respectively.

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