• Title/Summary/Keyword: 단일 효소 반응

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Biochemical Characteristics of Spodoptera exigua Nuclear Polyhedrosis Virus (파밤나방 핵다각체병 바이러스의 생화학적 특성)

  • 진병래;박범석;재연호;강석권
    • Korean journal of applied entomology
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    • v.30 no.2
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    • pp.144-149
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    • 1991
  • Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

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Purification and Characterization of Polyphenol Oxidase from Flammulina velutipes (팽나무버섯 polyphenol oxidase의 정제 및 특성)

  • Pyo, Han-Jong;Son, Dae-Yeul;Lee, Chan
    • Korean Journal of Food Science and Technology
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    • v.34 no.4
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    • pp.552-558
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    • 2002
  • Polyphenol oxidase from Flammulina velutipes was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Superdex G-200 gel filtration chromatography, Phenyl superose affinity chromatography, Mono-Q anion exchange chromatography and Superdex S-200 gel filtration chromatography on FPLC. After these purification steps specific activity of purified polyphenol oxidase increased to 199.1 units/mg. Polyphenol oxidase from F. velutipes was composed of a single polypeptide with molecular weight of about 40 kDa. Optimum pH and temperature for the enzyme reaction were found to be 6.0 and $25^{\circ}C$, respectively. The activity of the enzyme gradually decreased at acidic pH between 3 and 5, and the enzyme lost its activity at alkaline pH between 8 and 10. This enzyme exhibited high substrate specificity to o-diphenols. Km-values for L-DOPA and caffeic acid were found to be 3.97 mM and 1.78 mM, respectively. 2-mercaptoethanol, L-ascorbic acid, sodium bisulfite, EDTA and $Mg^{2+}$ inhibited the activity of pholyphenol oxidase and $Cu^{2+}$, $Fe^{2+}$, $Zn^{2+}$ and $Ni^{2+}$ increased enzyme activity. The activity of enzyme was well maintained at $-70^{\circ}C$ for over 4 months, and at $-20^{\circ}C$ for 1 months.

Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance. ((β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성)

  • Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Min-Jeong;Lee, Young-Choon;Lee, Jai-Heon;Joo, Woo-Hong;Choi, Yung-Hyun;Lim, Hak-Seob;Kim, Jeong-In;Seo, Kwon-Il;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.845-851
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    • 2008
  • ${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

진공동결건조 후 두부의 수분활성 및 품질변화에 관한 연구

  • 김진성;이준호;하영선
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.163.1-163
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    • 2003
  • 식품의 건조는 저장성을 향상시키고 수송을 간편하게 하나 특별한 경우는 건조과정에서 일어나는 성분변화에 의해 풍미, 색깔, 조직 등이 향성되게 하는 경우도 있다. 그러나 일반적으로 건조에 의해 풍미의 저하, 색깔의 퇴조, 영양성분의 손실 및 조직과 형태의 손상 등 품질을 저하시키므로 가능한 저온에서 단시간에 수분을 제거해야 한다. 건조식품의 저장성은 수분활성도, 제품의 종류, 저장온도등 다양한 요인들에 영향을 받으며, 특히 수분 활성도에 따라 비효소 갈색화 반응, 지방의 산패, 미생물의 발생정도가 달라진다. 따라서 건조식품 저장시 변질을 방지하기 위해서는 식품의 수분함량을 측정할 수 있는 등온흡습곡선을 작성하는 것이 중요하다. 또한 식품의 단분자층 수분함량을 결정하고, 흡습에 필요한 에너지를 구함으로서 건조식품의 저장조건 및 포장조건의 선택시 유용한 자료가 된다. 특히 식품분말의 흡습특성은 분말입자의 크기, 형태, 화학성분 등과 밀접한 관련이 있다. 위의 사실로 미루어 볼 때 본 연구에서는 시판 유통되고 있는 두부를 진공동결 시킴으로써 위생화와 장기저장 가능성을 연구하면서 진공동결 건조 후 시료의 수분활성, 재수화 및 일반성분 변화를 검토하고자 한다.

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Detection of Cucumber Mosaic Virus by RT-PCR Using a Simple and Rapid Crude Sap Extraction Method (간이 조즙액 추출법을 이용한 RT-PCR 방법에 의한 오이 모자이크 바이러스의 검정)

  • 이상용;홍진성;이진상;최장경
    • Korean Journal Plant Pathology
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    • v.12 no.4
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    • pp.432-436
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    • 1996
  • 역전사 중합효소 연쇄반응(RT-PCR)을 이용하여 담배(Nicotiana glutinosa)에 증식시킨 7종의 오이 모자이크 바이러스(CMV)를 검정하였다. RT-PCR을 위한 간단하고 신속한 바이러스 핵산의 조즙액 추출법이 개발되었으며, CMV 외피단백질 유전자 부위를 기초로 하여 제작한 20개의 염기로 구성된 primer를 사용하여 RT-PCR을 실시한 결과, 약 490 염기쌍의 DNA 단편들이 이병식물의 조즙액으로부터 증폭되었다. EcoRI 및 MspI을 이용한 RT-PCR 산물의 분석에 의하여, 공시한 7종의 바이러스는 모두 CMV subgroup I으로 동정되었다. Ouchterlony 한천젤 이중 확산법을 이용한 항혈청 검정에서도 7종의 바이러스 모두 CMV-Y의 항혈청과 단일의 침강선을 형성하였다.

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Identification of Introduced Gene and Its Expression and Gene Stability Assessment for Event Selection of Genetically Modified Plant toward Approval: Cucumber Mosaic Virus Resistant Hot Pepper (상업용 유전자 변형작물 이벤트 선발을 위한 도입유전자 확인, 발현 및 세대간 안정성 평가 : 오이모자이크바이러스 저항성 GM 고추)

  • Kang, Seung-Won;Han, Bal-Kum;Lee, Tae-Ho;Kim, Eun-Ji;Lee, Gung-Pyo
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.192-200
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    • 2012
  • For the development of genetically modified plants, it is important to verify various factors which potentially affect the risk assessment as well as to establish an experimental program to produce scientific and reliable data. However, it is a time and cost consuming process to develop GM plants as well as to prepare scientific and convincible data for government's approval. Therefore, using the transgenic hot pepper tolerant to a new CMV pathotype, we attempted to suggest few methodological procedures, such as probe saturation for southern blot analysis and RT-PCR and ELISA for expression analysis, for identification and stability evaluation of inserted gene in genetically modified plant which are required for submission for approval. Ten partially overlapped probes covering full length of inserted gene were produced. We could identify that the inserted gene was stacked as a single copy as well as no partial element existed. Also, we could identify the stability of the inserted gene stacked in hot pepper using probe saturation. In the expression analysis with RT-PCR and ELISA, we also could provide the stable expression of transcript and proteins in leaves and placenta and pericarp of fruits of the CMV-resistant hot pepper.

Preparation of Trinitrophenyl Cellulose as Substrate for Cellulase Assay (Trinitrophenyl Cellulose의 조제)

  • Maeng, Jeong-Seob;Nam, Yoon-Kyu;Park, Seung-Heui;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.22 no.2
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    • pp.151-155
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    • 1995
  • Two types of modified celluloses which contain trinitrophenyl groups as chromophore were synthesized from carboxymethyl cellulose Whatman CM 70 and CM 32. Diaminoethyl groups were added to the CM 70 and CM 32 to make DAE-CM celluloses and then the DAE-CM groups were substituted by 2,4,6-trinitrophenyl groups to produce TNP-celluloses. Average particle size of the TNP-cellulose from CM 32 was $44.6{\pm}9.6{\mu}m$ in diameter and $127.9{\pm}22.5{\mu}m$ in length, which was much smaller than those from CM 70, however its TNP-moiety per gram determined by using the molar extinction coefficient $1.33{\times}10^4$ of ${\varepsilon}$-TNP-lysine at 345 nm, was 0.68 millimoles, which was 5.6-fold greater than those from CM 70. The absorption spectrum of TNP-oligosaccharides which were the soluble products of TNP-celluloses by a cellulase preparation Onozuka R-10, showed a maximal peak at 344 nm. Increases in the absorbance during hydrolysis were linear with the enzyme concentration, and the differences of slope values between two types of TNP-celluloses that the more semsitive assay could be achieved by using those from CM 32 as substrate at the low range of the enzyme concentration.

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Two-step Acid Hydrolysis Method for Producing Fermentable Sugar from Lignocellulosic Biomass (발효당 생산을 위한 목질계 바이오매스의 2단 산당화)

  • Park, Jang Han;Kim, Jun Seok
    • Korean Chemical Engineering Research
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    • v.54 no.1
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    • pp.1-5
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    • 2016
  • For obtain fermentable sugar, we conducted acid hydrolysis with lignocellulosic biomass without enzyme. The lignocellulosic biomass used pinus rigida and Palm residues (EFB; empty fruit bunches). In the acid hydrolysis, we consider the hydrolysis condition to reduce a denatured sugar. So this study was conducted 2-step acid hydrolysis. First-step hydrolysis used high concentration (72 wt%) sulfuric acid at $80^{\circ}C$. At the condition, we obtained 11.49 wt%, 32 wt% glucose conversion for pinus rigida and EFB. After the step, the liquor was dilute until 9~15 wt% acid concentration and conducted second hydrolysis at $50{\sim}120^{\circ}C$. In the second hydrolysis, we obtained maximum glucose conversion (pinus rigida 86.8 wt% (39 g/L) and EFB 95.3 wt% (32.4 g/L)) at 9 wt% acid concentration and $120^{\circ}C$ for 80 min. All samples through the process are analyzed on the basis of mass balance.

Purification and Characterization of Cyclodextrin Glucanotransferase Excreted from Newly Isolated Alkalophilic Bacillus circulans (Alkalophilic Bacillus circulans가 생산하는 Cyclodextrin Glucanotransferase 의 정제와 효소반응특성)

  • 신현동;이상호;이용현
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.370-378
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    • 1989
  • An Alkalophilic Bacillus circulans that can produce significant amount of cyclodextrin glucanotransferase (CGTase) was newly isolated from soil. The culture filtrate was successively purified by ($NH_4$)$_2$$SO_4$precipitation, DEAE-Sephadex column chromatography, and Sephadex G-100 column chromatography. The enzymatic properties, including molecular weight, optimal pH and temperature, stability, and kinetic parameters, were determined. The cyclodextrin synthesis reaction catalized by the purified CGTase was also studied. The sweet potato and corn starch were found to be the most suitable substrates with 60% conversion to cyclodextrin. The highest conversion was achieved at the CGTase concentration of 900-1,100 units/g of soluble starch. The purified CGTase could also catalize the transglycosylation on stevioside.

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Porduction and Enzymatic Characteristics of ${\alpha}-Galactosidase$ from Aspergillus niger (Aspergillus niger에 의한 ${\alpha}-Galactosidase$의 생산 및 효소적 특성)

  • Chun, Hyang-Sook;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.20 no.1
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    • pp.79-84
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    • 1988
  • ${\alpha}$-Galactosidase from Aspergillus niger as a possible enzyme for removal of flatulence factors in soybean foods was produced the highest in 120 hours in either Czapeck-Dox liquid medium or wheat bran solid medium. The most efficient carbon and nitrogen sources in Czapeck-Dox medium were raffinose and sodium nitrate, respectively, whereas the addition of the sources showed negative effects in wheat bran. pH optima for enzyme activity and stability were 4.0-5.0 and 3.5-6.5, respectively, and optimum temperature for stability was $40-50^{\circ}C$. Upon reaction on p-nitrophenyl-${\alpha}$-D-galactoside, Michaelis constant was 0.42 mM and maximum velocity was 152 ${\mu}moles$ substrate/minute/kg solid medium. Mercuric chloride acted as a strong noncompetitive inhibitor and p-chloromercuribenzoate, even in low concentration, acted as a competitive inhibitor. Crude ${\alpha}$-galactosidase hydrolyzed raffinose and stachyose completely, giving spots of monosaccharides only on thin-layer chromatogram.

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