• 제목/요약/키워드: StyB

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Production, Purification, and Characterization of Soluble NADH-Flavin Oxidoreductase(StyB) from Pseudomonas putida SN1

  • Yeo, Yun-Ji;Shin, Seung-Hee;Lee, Sun-Gu;Park, Sung-Hoon;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
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    • 제19권4호
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    • pp.362-367
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    • 2009
  • In recombinant strains, many proteins and enzymes are expressed as inactive and insoluble inclusion bodies. For soluble expression of an active form of StyB, an NADH-flavin oxidoreductase, several recombinant Escherichia coli strains were developed and tested. Among them, strain BL21(DE3)pLysS effectively produced an active and soluble form of StyB as about 9% of the total protein content, when cultivated at $20^{\circ}C$ with 0.5 mM IPTG. The solubly expressed StyB has the highest oxidoreductase activity at pH 6.5-7.5 and $37^{\circ}C$. Substrate dependence profiles of the StyB-catalyzed reaction showed that the maximum specific activity($V_m$) and half saturation constant($K_m$) were $1,867{\pm}148\;U/mg$ protein and $51.6{\pm}11{\mu}M$ for NADH, and $1,274{\pm}34\;U/mg$ protein and $8.2{\pm}1.2{\mu}M$ for FAD, respectively. This indicates that solubly produced StyB has 6- to 9-fold higher oxidoreductase activities than the in vitro refolded StyB from inclusion bodies.

Characterization of Styrene Catabolic Genes of Pseudomonas putida SN1 and Construction of a Recombinant Escherichia coli Containing Styrene Monooxygenase Gene for the Production of (S)-Styrene Oxide

  • Park Mi-So;Bae Jong-Won;Han Ju-Hee;Lee Eun-Yeol;Lee Sun-Gu;Park Sung-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제16권7호
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    • pp.1032-1040
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    • 2006
  • Some Pseudomonas species can grow on styrene as a sole carbon and energy source. From the new isolate Pseudomonas putida SN1, the genes for styrene catabolism were cloned and sequenced. They were composed of four structural genes for styrene monooxygenase (styA and styB), styrene oxide isomerase (styC), and phenylacetaldehyde dehydrogenase (styD), along with two genes for the regulatory system (styS and styR). All the genes showed high DNA sequence (91% to 99%) and amino acid sequence (94% to 100%) similarities with the corresponding genes of the previously reported styrene-degrading Pseudomonas strains. A recombinant Escherichia coli to contain the styrene monooxygenase from the SN1 was constructed under the control of the T7 promoter for the production of enantiopure (S)-styrene oxide, which is an important chiral building block in organic synthesis. The recombinant E. coli could convert styrene into an enantiopure (S)-styrene oxide (ee >99%) when induced by IPTG The maximum activity was observed as 140 U/g cell, when induced with 1 mM IPTG at $15^{\circ}C$.

담배가루이 Biotype B와 Q의 약제감수성과 효소활성 비교 (Comparison of insecticide susceptibility and enzyme activities of biotype B and Q of Bemisia tabaci)

  • 김은희;성재욱;양정오;안희근;윤창만;서미자;김길하
    • 농약과학회지
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    • 제11권4호
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    • pp.320-330
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    • 2007
  • 충북 진천군 이월면 장미재배지와 경남 밀양시 고추밭에서 채집한 담배가루이에 대해서 16S DNA 염기서열을 분석하였다. 각각의 PCR 산물을 EcoT14 I (Sty I)과 Stu I을 처리한 결과 경남 밀양시 고추밭에서 채집한 담배가루이는 Sty I 에서 555pp와 311bp 두 개의 단편이 만들어져 biotype Q임을 확인하였고, 충북진천군 장미재배지에서 채집한 담배가루이는 Stu I에서 560bp와 3060p 두 개의 단편이 만들어서 biotype B임을 확인하였다. 두 biotype에 대한 12종 살충제로 발육단계별 약제감수성, 침투이행성 및 잔효성을 비교하였고, esterase, acetylcholinesterase (AChE), glutathione S-transferase 등의 효소활성에 미치는 저해정도를 검토하였다. 두 biotype간에 약제감수성 차이가 있었고, biotype B가 Q보다 더 감수성이었다. 엽면침투이행성과 근부침투이행성 및 잔효성에서도 biotype B가 Q보다 더 감수성이었다. 유기인계인 fenitrothion과 카바메이트계인 fenothiocarb의 저해제에 대해서 esterase, acetylcholinesterase (AChE), glutathione S-transferase등의 효소활성에 미치는 영향을 검토한 결과, biotype Q가 B보다 활성이 높게 나타났다. 이상의 결과를 종합하여 볼 때 담배가루이 biotype Q가 B보다 약제에 대해서 저항성임을 알 수 있었다.

플라즈마중합 (MMA-Sty-TMT) 박막의 레지스트 특성조사 (A study on the resist characteristics of plasma polymerized thin film of (MMA-Sty-TMT))

  • 박종관;박상현;박복기;정해덕;한상옥;이덕출
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 1994년도 하계학술대회 논문집 C
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    • pp.1268-1270
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    • 1994
  • Fine lithographic technology in a submicron design regime is necessary for the fabrication of VLSI circuits. In such lithography, fine pattern delineation is performed by electron beam, ion beam and X-ray lithography instead of photolithography. Therefore, the new resist materials and development method have been required. So, we are investigating another positive E-beam resists which have high sensitivity and dry etching resistance, Plasma co-polymerized resist was prepared using an interelectrode gas-flow-type reacter. Methymethacrylate, tetramethyltin and styrene were chosen as the monomer to be used. The delineated pattern in the resist was developed with gas-flow-type reactor using an argon and 02 as etching gas. We studied about the effects of discharge power and mixing rate of the co-polymerized thin :film. The molecular structure of thin film was investigated by ESCA and IR, and then was discussed in relation to its quality as a resist.

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DNA 다형(多型)에 있어서 진도견(珍島犬)과 잡종견(雜種犬)과의 비교(比較) (Polymorphism of mitochondrial DNA in Jindo dogs and Japanese mongrels dogs)

  • 한방근;김주헌;강주원;이케모토 시게노리
    • 대한수의학회지
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    • 제33권1호
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    • pp.43-51
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    • 1993
  • Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

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Cell Viability in $G_0$-like Stationary Phase of Schizosaccharomyces pombe: Roles of Psp1/Sds23 and Ufd2

  • Jang, Young-Joo;Ji, Jae-Hoon;Chung, Kyung-Sook;Kim, Dong-Uk;Hoe, kwang-Lae;Won, Mi-Sun;Yoo, Hyang-Sook
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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    • pp.110-113
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    • 2005
  • Under the condition of nutritional deprivation, actively growing cells prepare to enter $G_0$-like stationary phase. Protein modification by phosphorylation/dephosphorylation or ubiqutination contributes to transfer cells from active cell cycle to dormant stage. We show here that Psp1/Sds23, which functions in association with the 20S cyclosome/APC (1) and is essential for cell cycle progression in Schizosaccharomyces pombe (2), is phosphorylated by stress-activated MAP kinase Sty1 and protein kinase A, as well as Cdc2/cyclinB, upon entry into stationary phase. Three serines at the positions 18,333 and 391 are phosphorylated and overexpression of Psp1 mutated on these sites causes cell death in stationary phase. These modifications are required for the binding of Spufd2, a S.pombe homolog of multiubiquitin chain assembly factor E4 in ubiquitin fusion degradation pathway. Deletion of Spufd2 gene led to increase cell viability in stationary phase, indicating that S. pombe Ufd2 functions to inhibit cell growth at this stage to maintain cell viability. Moreover, Psp1 enhances the multiubiquitination function of Ufd2, suggesting that Psp1 phosphorylated by sty1 and PKA kinases is associated with the Ufd2-dependent protein degradation pathway, which is linked to stress tolerance, to maintain cell viability in the $G_0$-like stationary phase.

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관광호텔 이용객의 음식선택요인에 관한 연구 -한식, 중식, 일식을 중심으로- (A Study on Food Choice Factors of Tourist Hotel Customers′)

  • 유택용
    • 한국조리학회지
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    • 제6권2호
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    • pp.97-122
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    • 2000
  • It is currently that there are increasing especially of F&B's revenue well as room's. Thus, Hotel have set foods adapt to customers' need. This paper is targeting on figuring out for food choice factors of tourist hotel customers. Results on this are 1ike follows; Firstly, major factors on the food choice are different by its style respectively. \circled1 Menu's diversity and nutrition degrees have highly influence with Korean sty1e. \circled2 Freshness, Food's degree of temperature and appearance have highly influence with Japanese style. \circled3 Nutritious balance and its appearance have highly influence Chinese style. Secondly, rest factors on the food choice are shown that service factor is generally important in these three style.

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Enterovirus에 대한 분자생물학적 검증법 및 Genotypes 방법의 개발 (The Development of Molecular Detection Method and Differentiation of Genotypes of Enterovirus)

  • 김은순;남정현;김기순;윤재득;김유겸
    • 대한바이러스학회지
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    • 제27권2호
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    • pp.169-176
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    • 1997
  • In this study, the feasibility of identification and genotypic differentiation of enteroviruses was investigated by using nested reverse transcription-polymerase chain reaction (nested RT-PCR), single-stranded conformation polymorphism (SSCP), and restriction fragment length polymorphism (RFLP) techniques. Two hundred seventy-four clinical samples were assayed by both nested RT-PCR and tube culture method using MRC-5 and MK cells; 58 (86.6%) out of 67 enterovirus culture-positive samples contained enteroviral RNA. In addition, 114 (55.1%) of 207 samples from patients with suspected enteroviral CNS disease with negative viral cultures were positive by the nested RT-PCR. The nested RT-PCR products were genotyped by the SSCP method and the results were compared with serotypes. We could differentiate 6 subtypes, 3 of which are similar to coxsackievirus B3, B5, echovirus 11, plus 3 other subtypes. RFLP cleaved with Sty I, Bgl I, and Xmn I yielded characteristic patterns for each laboratory strains. This study demonstrates the usefulness of the RT-PCR for the rapid diagnosis of enterovirus infection and the potentials of the SSCP method for differentiation of enterovirus strains.

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간호과장의 지도성 유형과 병원 조직풍토와의 관계 (A Study on the Relationships Between the chief Nurse′s Leadership style and the organizational climate of Hospitals)

  • 김영매;한상임
    • 대한간호학회지
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    • 제5권1호
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    • pp.79-86
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    • 1975
  • It had been thought the role of Nursing administrator was still not started along the right line in Korea because of the Lacking of full understanding and recognition of the real meaning of administrator's role, and because there are several Rinds of problems and difficulties in actual role. The present study is an attempt to clarify some existing relationships between the chief Nurse's Leadership style and the organizational climate of Hospitals. The problems of the study are specifically started as follows ; 1) What influence does the individual behavior have on the formation of the organizational climate of Hospital? 2) From what do the difference of climate arise? 3) How are the individual be heavier and organizational climate of Hospital measured ? In order to see the relationships or the interactions between the two factors, the chief nurse's Leadership style and organizational climate of hospital , the researcher has classified. He former into; 1) effective Leadership style. 2) initiating structure leader ship style. 3) Consideration structure Leadership style, and 4) ineffective Leadership style, and the latter in to. 1) open climate and 2) closed climate The Chief Nurses Leadership style has been. Classified into consideration-human relations approaches and initiating structure approaches according to L. B. D. Q by Hapin. Organizational climate of hospital has been classified into open-closed continuum according to O. C. H Q. by Hairpin and Croft. The results obtained are as follows : 1) The chief nurse's Leadership style is closer to Initiating structure Leadership style than Consideration structure lure Leadership style. 2) The organizational climate of hospital is closer to open climate than closed climate. 3) The chief nurse's Leadership sty]e and the organizational climate of hospital to not show any significant relations.

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