• 제목/요약/키워드: Immobilization technique

검색결과 90건 처리시간 0.048초

Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

  • Misbah, Asmae;Koraichi, Saad Ibnsouda;Jouti, Mohamed Ali Tahri
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.491-505
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    • 2020
  • A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

The Covalent Immobilization of Biomolecules to Polymer Surface by Deep-UV Lithography Using N-Hydroxysuccinimidyl Azidobenzoate

  • 김현정;;;;민병구
    • 대한의용생체공학회:학술대회논문집
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    • 대한의용생체공학회 1997년도 춘계학술대회
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    • pp.47-50
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    • 1997
  • We synthesized N-Hydroxysuccinimidyl (NHS) azidobenzoate as a cross-linking reagent for immobilization of peptide onto the solid surface. Thin polystyrene(PS) films spin-coated with a NHS azidobenzoate solution were exposed with ultraviolet light at 245nm$(3.3mW/cm^2)$ for 5 min. The NHS active ester groups became covalently attached to the polymer via photogenerated, highly reactive nitrene intermediates derived from NHS azidobenzoate. Using this technique, it is demonstrated that well-defined surface regions can be functionalized with a minimum observable feature size of 1mm for UV exposure. Through reaction of this functionalized PS surface with primary amine-containing biomolecules, biological molecule had been immobilized on the polymer surface.

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Picomolar Scale Determination of Carbohydrates Covalently Immobilized on Activated Beads Using Hydroxyl Functionality

  • Yu, Jae-Hoon;Chun, Sung-Min;Park, Ho-Koon;Park, Yong-Keun;Jeong, Sun-Joo
    • BMB Reports
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    • 제32권1호
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    • pp.98-102
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    • 1999
  • Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

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전기 화학 응용을 위한 폴리옥소메탈레이트와 나노물질의 나노하이브리드화 (Nanohybridization of Polyoxometalate and Nanomaterials for Electrochemical Application)

  • 양민호;최봉길
    • 공업화학
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    • 제29권4호
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    • pp.363-368
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    • 2018
  • Polyoxometalates (POMs)는 뛰어난 특성과 전기 화학 응용 분야에 대한 많은 잠재력을 가지고 있다. POM은 매우 잘 녹는 성질 때문에 전기화학 소자에서 POM의 잠재력을 최대한 활용하기 위해서는 다양한 기능성 재료에 POM을 고정화하는 과정이 필수이다. 본 논문에서는 우리는 최근 개발된 고정화 방법인 나노 카본 및 전도성 고분자와 같은 전도성 나노 물질에 POM을 도입하는 기술들에 대해서 논하고자 한다. Langmuir-Blodgett 기술, 층별 자기 조립 및 전기화학 in-situ 중합을 사용하여 전도성 고분자 매트릭스 및 POM을 나노 카본으로 도입할 수 있는 다양한 고정화 전략을 소개한다. 또한 우리는 POM의 응용 분야인 물 산화용 전극 촉매, 리튬 이온 배터리, 슈퍼커패시터 및 전기화학적 바이오 센서 등의 다양한 전기 화학 응용 분야를 다룬다.

소수성 상호작용을 이용한 고집적 DNA칩 마이크로어레이의 개발 (Development of High-Intergrated DNA Chip Microarrays by Using Hydrophobic Interaction)

  • 김도균;최용성;권영수
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 하계학술대회 논문집
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    • pp.757-760
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    • 2001
  • We have used the random fluidic self-assembly (RFSA) technique based on the chip pattern of hydrophobic self-assembly layers to assemble microfabricated particles onto the chip pattern. Immobilization of DNA, fabrication of the particles and the chip pattern, arrangement of the particles on the chip pattern, and recognition of each using DNA fluorescence measurement were carried out. Establishing the walls, the arrangement stability of the particles was improved. Each DNA is able to distinguish by using the lithography process on the particles. Advantages of this method are process simplicity, wide applicability and stability. It is thought that this method can be applicable as a new fabrication technology to develop a minute integration type biosensor microarray.

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Quantitative Evaluation of Setup Error for Whole Body Stereotactic Radiosurgery by Image Registration Technique

  • Kim, Young-Seok;Yi, Byong-Yong;Kim, Jong-Hoon;Ahn, Seung-Do;Lee, Sang-wook;Im, Ki-Chun;Park, Eun-Kyung
    • 한국의학물리학회:학술대회논문집
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    • 한국의학물리학회 2002년도 Proceedings
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    • pp.103-105
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    • 2002
  • Whole body stereotactic radiosurgery (WBSRS) technique is believed to be useful for the metastatic lesions as well as relatively small primary tumors in the trunk. Unlike stereotactic radiosurgery to intracranial lesion, inherent limitation on immobilization of whole body makes it difficult to achieve the reliable setup reproducibility. For this reason, it is essential to develop an objective and quantitative method of evaluating setup error for WBSRS. An evaluation technique using image registration has been developed for this purpose. Point pair image registrations with WBSRS frame coordinates were performed between two sets of CT images acquired before each treatment. Positional displacements could be determined by means of volumetric planning target volume (PTV) comparison between the reference and the registered image sets. Twenty eight sets of CT images from 19 WBSRS patients treated in Asan Medical Center have been analyzed by this method for determination of setup random error of each treatment. It is objective and clinically useful to analyze setup error quantitatively by image registration technique with WBSRS frame coordinates.

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고정화된 Streptomyces lincolnensis의 반복 회분식 배양에 의한 린코마이신 생산 (The Production of Lincomycin by Repeated Batch Cultures of Immobilized Streptomyces lincolnensis)

  • 김창준;전계택;장용근;김성배
    • KSBB Journal
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    • 제21권5호
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    • pp.384-388
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    • 2006
  • 본 연구에서는 고정화 방선균을 이용한 이차대사산물생산 공정 개발에 있어서 고려해야 할 중요한 사항인, 고정화세포의 반복사용 가능성을 조사하였다. 또한 축축한 담체에 균사체 형태의 생산균주의 고정화가능성에 대하여 알아보았다. 셀라이트 담체에 고정화된 고생산성 Streptomyces lincolnensis 돌연변이주를 이용한 반복회분식 배양에서, 회분수가 증가함에 따라 고정화세포의 린코마이신 생산성이 증가하였고 9번째 회분에서는 1007 $({\pm}256)$ mg/L의 린코마이신이 얻어졌다. 10번의 반복회분동안 고정화세포에 의해 얻어진 린코마이신 총양을 기준으로 계산된 고정화세포 배양의 생산성은 현탁회분식 배양에 비해 1.4배 높았다. 축축한 담체와 배지가 포함된 플라스크에 균사체 형태의 생산균주를 접종하고 배양 후 생성된 고정화세포 농도와 린코마이신 양은 건조한 담체에 포자상태의 생산균주를 고정화한 후 배양하여 얻어진 것에 비하여 다소 높음이 관찰되었다. 이는 균사체 형태의 생산균주가 축축한 담체에 쉽게 고정화되는 것을 의미한다. 실제 생산규모를 고려할 때, 본 연구팀에서 개발한 방선균 고정화방법은 담체 멸균 및 세포고정화 단계에서 추가적인 설비가 필요 없으며, 이는 고정화단계에서 숙련된 기술과 추가의 설비를 요하는 다른 방법들에 비하여 실용적이며 경쟁력 있는 생물 공정이라고 사료된다.

양어장수내의 암모니아성 질소제거를 위한 질화세균군의 고정화 (Immobilization of Nitrifier Consortium for the Removal of Ammonium Ion in the Recirculating Aquaculture System)

  • 김성구;서재관;이종석;공인수;서근학
    • 한국수산과학회지
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    • 제30권5호
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    • pp.816-822
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    • 1997
  • 최근 biotechnology 발전과 같이하여 미생물 고정화법이 급속히 진보하였다. 도처에 있는 biological midia로부터 products를 추출하기 위해 미생물 고정화법은 많이 적용되고 있지만 아직 하수처리에 적용은 되고 있지 않다. 그러므로 본 연구에서는 순환 양식 시스템에 고정화기술의 적용 가능성을 타진하였다. fludized bed reactor에서 bead에 고정화된 질화세균군을 사용하기 위해서는 고정화 bead의 내구성이 높아야함으로 각 고정화법에 따른 강도는 $Ba^{++}-alginate$ bead가 1450g의 breaking force로 가장 높은 내구력을 지니고 있음으로 가장 적합한 bead라고 할 수 있었다. 또한 20 mg/L ammonium ion을 제거 하는데 $Ba^{++}-alginate$ bead는 다른 두 bead보다 훨씬 논은 활성을 나타내었으며, 아울러 고정화 질화세균군이 비고정화 질화세균군보다 높은 질화 효율을 보여 주었다.

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치위생과 학생의 치근단 촬영법 인식에 관한 조사 연구 (A survey study on recognition of periapical radiography in dental hygiene students)

  • 박일순;정정옥;이경희
    • 한국치위생학회지
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    • 제12권5호
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    • pp.987-997
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    • 2012
  • Objectives : This study was carried out in order to obtain basic data for students' efficient acquirement and instruction of radiography technology in the future by surveying dental hygiene students' recognition of periapical radiography. Methods : This study carried out a questionnaire survey targeting dental hygiene students from December 2009 to December 2010, and obtained the following results. Results : 1. As a result of examining recognition on periapical radiography, the bisecting angle technique was indicated to be averagely $3.84{\pm}0.566$ points. The paralleling technique was indicated to be $2.66{\pm}0.701$ points. 2. As a result of examining about problems given the bisecting angle technique, what had been most difficult given the bisecting angle technique was indicated to be the highest in cone positioning with 34.2%. The most difficulty given deciding on the X-ray vertical-angel irradiation direction was indicated to be the highest with 66.9% in adjusting the cone direction on the virtual bisector. 3. As a result of examining about problems given the paralleling technique, what had been most difficult in the process of the paralleling technique was indicated to be the highest with 56.7% in fixing the film immobilization device inside the mouth. Conclusions : Examining the above results, it is considered that there is a need of understanding morphological and anatomical structure inside the mouth in order to reduce the mistake rate given the periapical radiography, and that it is important to increase skill level by repetitively shooting several times with having enough time.

공유결합으로 고정화된 urokinase 칼럼의 스케일업과 solid-phase refolding에 의한 반복 사용

  • 서창우;안상점;이은규
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.85-88
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    • 2001
  • hGH 와 GST 절편으로 구성된 융합 단백질의 고정화 UK 칼럼에 의한 절단 반응 후 용출액의 pH를 3.5 로 낮춤으로써 이물질들을 침전시키고 이를 expanded bed chromatography 칼럼에 통과시킨 결과, 이물질들의 제거와 hGH 단량체의 흡착분리가 동시에 이루어겼다 . 흡착된 단량체는 NaCl 에 의해 용출되었으며 이 단계의 수율은 거의 100% 이었다 , 따라서 칼럼에 의한 절단 반응과 산 침전에 의한 이물질 침잔 반응 EBA 에 의한 이물질 제거 및 단량체 회수 반응을 연속적으로 진행할 수 있는 기초를 제시하였다 . 또한 고정화된 UK 는 guanidine HCl(6M)을 이용하여 unfolding 시키고 이를 세척하여 refolding 시킨 결과 20 회의 반복적인 처리 후에도 초기 활성의 약 80% 수준을 유지하였다. 이는 UK 가 공유결합된 상태에서 solid-phase refolding 이 가능하다는 증거이며, 고정화 효소 칼럼의 수영을 크게 향상시켜 경제성을 확보하는 방안으로 이용될 것으로 기대된다.

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