• Title/Summary/Keyword: Immobilization method

Search Result 315, Processing Time 0.025 seconds

Production, Immobilization, and Characterization of Croceibacter atlanticus Lipase Isolated from the Antarctic Ross Sea (남극 로스해에서 분리한 Croceibacter atlanticus균 유래 리파아제의 생산, 고정화, 효소특성 연구)

  • Park, Chae Gyeong;Kim, Hyung Kwoun
    • Microbiology and Biotechnology Letters
    • /
    • v.46 no.3
    • /
    • pp.234-243
    • /
    • 2018
  • The Antarctic Ocean contains numerous microorganisms that produce novel biocatalysts that can have applications in various industries. We screened various psychrophilic bacterial strains isolated from the Ross Sea and found that a Croceibacter atlanticus strain (Stock No. 40-F12) showed high lipolytic activity on a tributyrin plate. We isolated the corresponding lipase gene (lipCA) by shotgun cloning and expressed the LipCA enzyme in Escherichia coli cells. Homology modeling of LipCA was carried out using the Spain Arreo lake metagenome alpha/beta hydrolase as a template. According to the model, LipCA has an ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Glymotif, and lid sequence, indicating that LipCA is a typical lipase enzyme. Active LipCA enzyme was purified fromthe cell-free extract by ammonium sulfate precipitation and gel filtration chromatography. We determined its enzymatic properties including optimum temperature and pH, stability, substrate specificity, and organic solvent stability. LipCA was immobilized by the cross-linked enzyme aggregate (CLEA) method and its enzymatic properties were compared to those of free LipCA. After cross-linking, temperature, pH, and organic solvent stability increased considerably, whereas substrate specificities did not changed. The LipCA CLEA was recovered by centrifugation and showed approximately 40% activity after 4th recovery. This is the first report of the expression, characterization, and immobilization of a C. atlanticus lipase, and this lipase could have potential industrial application.

Development of Protein Chip for Diagnosis of Chlamydophia Pneumoniae (단백질 칩을 이용한 클라미디아 폐렴의 진단)

  • Kim, Woo Jin;Lee, Hui Young;Lee, Seung-Joon;Jung, Se-Hui;Yuk, Jong Seol;Ha, Kwon-Soo;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
    • /
    • v.60 no.4
    • /
    • pp.412-418
    • /
    • 2006
  • Background; The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor. Methods; For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor. Results; The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens. Conclusion; The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field.

Effect of Yangsimtang and Yangsimtang + Siyup on the Regional Brain Catecholamines contents of Immobilization stessed Rats (양심탕(養心湯) 및 양심탕가시엽(養心湯加枾葉)이 구속(拘束)Stress 흰쥐의 뇌부위별(腦部位別) Catecholamines함량에 미치는 영향(影響))

  • Song Pil-Jung;Jeong Dae-Kyoo
    • Journal of Oriental Neuropsychiatry
    • /
    • v.8 no.1
    • /
    • pp.49-68
    • /
    • 1997
  • This study aimed to evaluate the anti-stress effect of Yangsimtang and Yangsimtang+Siyup on the rats in immobilization stress.The experimental animals were immobilized in the stress box(5${\times}$5${\times}$20cm) for 12 hours in a day suring 3 days, and administered 1g/100g of Yangsimtang and Yangsimtang+Siyup and Siyup extract for 12 days before stress. The norepinephrine, epinephrine, dopamine and serotonin contents were measured by HPLC method in various part of rat brain.The following results were observed.1. In frontal cortex the norepinephrine content of control group was 561.${\pm}$24.46 ng/g brain tissue, that of saple 1 group was 430.8$\pm$41.2 ng/g brain tissue, and that of sample 2 group was 417.2$\pm$38.5 ng/g brain tissue. The differences was statistically significant.2. In corpus striatum, the norepinephrine content of control group was 561.3$\pm$27.3 ng/g brain tissue, and that of sample 1 group was 422.1$\pm$21.2 ng/g brain tissue, the dopamine content of control group was 1205.1$\pm$75.9 ng/g brain tissue, that of sample 2 group was 685.6$\pm$41.5 ng/g brain tissue. The differences was statistically significant.3. In hypothalamus, the norepinephrine content of control group was 1165.1$\pm$162.6 ng/g brain tissue, that of sample 2 group was 947.2$\pm$35.7 ng/g brain tissue. The differences was statistically significant.4. In hippocampus, the norepinephrine content of control group was 931.6$\pm$82.2 ng/g brain tissue, that of sample 1 group was 652.1$\pm$47.5 ng/g brain tissue, and that of sample 2 group was 627.4$\pm$31.2 ng/g brain tissue, the dopamine contrnt of control group was 315.4$\pm$28.4 ng/g brain tissue, that of sample 2 group was 208.5$\pm$23.7 ng/g brain tissue. The differences were statistically significant.Base on the above results, it may be concluded that Yangsimtang and Yangsimtang+Siyup are effective to reduce stress.

  • PDF

Response of Photobacterium phosphoreum to Heavy Metal (Photobacterium phosphoreum의 중금속에 대한 반응성)

  • 정계훈;김현숙;이은수;정성제;이정건;김은기;전억한
    • KSBB Journal
    • /
    • v.14 no.3
    • /
    • pp.342-350
    • /
    • 1999
  • Photobacterium phosphoreum was used in order to study response to heavy metal including $HgCl_2$, $CdCl_2$, $MnSO_4$ and $ZnSO_4$ in view of developing monitoring system for toxic substances. The concentrations of heavy metal causing 50% reduction($EC_{50}$) in bioluminescence intensity were determined with both free and immobilized P. phosphoreum. The bioluminescence responses were examined at various concentrations of heavy metal after 10, 20 and 30 min of exposure. The linear correlation between Gamma values and concentrations of heavy metal was obtained and $EC_{50}$ was calculated from the linear correlation. The significant inhibitory concentrations for bioluminescence emission were found to be 0.05mg/L for $HgCl_2$, 25mg/L for $CdCl_2$, 50mg/L for $MnSO_4$ and 12.5mg/L for $ZnSO_4$, respectively. The free cell and disc type were shown to be more sensitive to heavy metal than cells mixed with Na-alginate or immobilized on Sr-alginate. However, the linear regression curves were derived from the Sr-alginate immobilized cells indicating the immobilization method is a useful tool for monitoring of heavy metal under more stable condition of bioluminescence.

  • PDF

Hydrolysis of Urea by Immobilized Urease Membrane (우레아제(Urease) 고정막에 의한 요소(Urea)의 가수분해)

  • Kim, Byoung-Sik;Kim, Min;Heo, Kwang-Beom;Hong, Joo-Hee;Na, Won-Jae;Kim, Jae-Hun
    • Applied Chemistry for Engineering
    • /
    • v.18 no.1
    • /
    • pp.10-16
    • /
    • 2007
  • In this study, we examined the preparation and hydrolysis property of immobilized urease membrane to decompose harmful urea in the body and remove ammonia which was produced by its decomposition. Urease immobilized membrane was prepared by introducing anion-exchange group DEA into porous hollow-fiber membrane by radiation graft polymerization method, and immobilization of urease. When urease was immobilized at membrane introduced with anion-exchange group, the more increasing grafting rate, the more increasing immobilization amount. The result originates from the fact that a greater amount of protein was immobilized by forming a multilayer on the longer grafted chain. Meanwhile, the addition of the cross-linker was possible not only to suppress separation phenomenon produced during a washing process of immobilized urease membrane but also to enable the recycling of membrane. Urease Immobilized membrane with no separation phenomenon was prepared by cross-linking reaction for 5 h, and the hydrolysis rate of prepared urease immobilized membrane was over 98% and 50%, respectively, in 1 mol and 4 mol urea solutions.

Polymerization of Ethylene over $Cp_2ZrCl_2$ Catalyst Supported on Montmorillonite (Montmorillonite에 담지된 $Cp_2ZrCl_2$ 촉매를 이용한 에틸렌 중합특성 연구)

  • Ahn, Sung-Hyun;Lee, Sung-Ho;Choi, Moo-Seok;Im, Jun-Seop;Sheikh, Rizwan;Cho, Deug-Hee;Park, Yeung-Ho
    • Applied Chemistry for Engineering
    • /
    • v.24 no.1
    • /
    • pp.55-61
    • /
    • 2013
  • Heterogeneous metallocene catalysts supported on montmorillonite (MMT), [$Cp_2ZrCl_2$/MMT, $Cp_2ZrCl_2$/MAO/MMT, and $Cp_2ZrCl_2$ + MAO/MMT], were prepared with three different methods of immobilization and tested for ethylene polymerization. The heterogeneous catalysts immobilized on organo clay (30B-MMT) showed the higher metal loading and polymerization activity than those immobilized on natural clay $Na^+-MMT$. These results suggest that the hydroxyl groups of organo clay interlayers react with the MAO and catalyst through the chemical bond. The metallocene catalyst supported directly on MMT showed lower activity for ethylene polymerization compared to the homogeneous systems, while MMT/MAO/$Cp_2ZrCl_2$, catalysts treated with MAO before impregnation, showed a higher activity. The polymers obtained from MMT-supported catalysts have higher melting point, molecular weight and molecular weight distributions than those of homogeneous catalysts. The polymer particles with increasing significant size. Ethylene polymerization over 30B-MMT/MAO/$Cp_2ZrCl_2$ catalyst was also performed varying the process variables to optimize the process conditions.

Synthesis of Cyclododecatriene from 1,3-Butadiene by Trimerization over Amine-Titanium Complex Catalyst (아민-티타늄착체 촉매상에서 1,3-부타디엔의 삼량화반응에 의한 싸이클로도데카트리엔의 합성)

  • Park, Da Min;Kim, Gye-Ryeong;Lee, Ju Hyun;Cho, Deuk Hee;Kim, Geon-Joong
    • Korean Chemical Engineering Research
    • /
    • v.51 no.3
    • /
    • pp.394-402
    • /
    • 2013
  • The new complex catalysts were synthesized by the reaction of titanium compounds (titanium chloride or titanium butoxide) and diamines in this work, and they showed very high catalytic activities for the cyclododecatriene (CDT) synthesis from 1,3-butadiene through trimerization. CDT synthetic reaction was performed in an autoclave reactor, and the effects of reaction temperature, type of catalyst, catalyst amount added into the system, the mole ratio of Al/Ti and immobilization method were investigated on the yield of product CDT. The titanium complex catalyst combined to diamine with 1:1 ratio showed high selectivity to CDT more than 90%. The ratio of TTT-CDT/TTC-CDT isomers in the product revealed as different values, depending on the type of diamine combined to titanium and Ti/diamine ratios. Those homogeneous complexes could be used as a heterogenized catalyst after anchoring on the supports, and the immobilized titanium catalyst retained the catalytic activities for several times in the recycled reactions without leaching. The carbon support containing titanium has exhibited superior activity to the silica support. Especially, when the titanium complex was anchored on the support which was fabricated by the hydrolysis of tripropylaminosilane itself, the resulting titanium catalyst showed the highest BD conversion and CDT selectivity.

The Critical Pigment Volume Concentration Concept for Paper Coatings: I. Model Coating Systems Using Plastic Pigments and Latex Binders for Paper Coating Applications

  • Lee, Do-Ik
    • Journal of Korea Technical Association of The Pulp and Paper Industry
    • /
    • v.34 no.5
    • /
    • pp.1-17
    • /
    • 2002
  • The immobilization and consolidation of the model coatings based on the plastic pigment and latex binder of known particle sizes were theoretically Studied in terms of the dense random packing of binary spheres and varying extent of latex film shrinkage. The porosity of the model coatings was calculated based on three proposed latex shrinkage models: Maximum, Minimum, and Linearly Decreasing Latex Shrinkage. The increasing extent of latex shrinkage was calculated up to the critical pigment volume concentration(CPVC) as a function of plastic pigment volume fractions, and the maximum latex shrinkage was estimated from the CPVC. Also, the number of pores and the average equivalent spherical pore diameters were calculated based on those proposed models. The opacity and gloss of the model coatings on polyester films were measured and their porosity was also determined by a simple coat weight-thickness method. As expected, various coating structure-property-composition relationships, such as opacity, gloss, porosity, etc., were shown to exhibit sharp transitions near the CPVC. The CPVC values determined by the opacity, gloss, and porosity vs. PVC relationships, respectively, agreed very well with each other. Especially, the CPVC's determined by the opacity and porosity vs. PVC curves were identical. The comparison between the theoretically calculated and experimental porosity values showed that the intermediate value between the maximum and minimum latex shrinkage would best fit the experimental porosity data. The effect of plastic pigment particle size on the optical properties and porosity of model coatings was also studied and it was observed that the coating opacity and porosity increased with increasing plastic pigment particle size, but the gloss decreased. The ink gloss of the uncalendered model coatings applied onto commercial sheet offset coated papers was shown to be affected by both the coating gloss and porosity: the higher the coating gloss, the higher the ink gloss, but the higher the coating porosity, the lower the ink gloss. Their printability was also studied in terms of the number of passes-to-fail and the rate of ink setting as a function of both plastic pigment volume fractions and plastic pigment particle sizes. A minimum crack-free temperature(MCR) of latex-bound coatings was proposed to better predict the behaviors of latexes as coating binders. The wet state of model coating dispersions, the surfaces of consolidated model coatings, and their internal structure were examined by both electron and atomic force microscopy, and their micrographs were found to be consistent with our immobilization and consolidation models.

Immobilization Effect and Abdominal Pressure of Newly-Developed Lumbosacral Spinal Orthosis during Task Performance (새롭게 개발한 요천추 보조기의 과제 수행 중 척추의 고정효과와 복부압력)

  • Jeon, Kyung Soo;Yang, Hee Seung;Jang, Soo Woong;Shin, Hee Dong;Lee, Yun kyung;Lee, Young;Lee, Seul Bin Na;Ahn, Dong Young;Sim, Woo Sob;Cho, Min;Cho, Kyu Jik;Park, Dong Beom;Park, Kwan Soo
    • Clinical Pain
    • /
    • v.19 no.2
    • /
    • pp.70-79
    • /
    • 2020
  • Objective: We evaluated the efficacy of a newly-developed spinal orthoses (V-LSO) by comparing the stabilizing effect, abdominal pressure, and comfort of 3 different semirigid LSOs (classic LSO, V-LSO, and Cybertech®) during various body movements. Method: Thirty healthy volunteers (23~47 years, 24 males, 6 females) were selected. A dual inclinometer measured the range of motion (ROM) while the participants performed flexion/extension and lateral flexion of the lumbar spine with 3 LSOs. The LSO's pressure on the abdominal surface was measured using 9 pressure sensors while lying, sitting, standing, flexion/extension, lateral flexion, axial rotation, and lifting a box. Comfort and subjective immobilization were analyzed by a questionnaire. Results: V-LSO had a statistically significant effect on flexion over Cybertech®. No significant differences were noted during extension and lateral flexion between the 3 LSOs. The abdominal pressure showed no significant differences while supine. While sitting, standing, and lifting a box, the mean abdominal pressure for V-LSO were significantly higher than those for Cybertech®. During lumbar flexion, the mean abdominal pressures for classic LSO and V-LSO were significantly higher than that of Cybertech®. For extension, lateral flexion and axial rotation, the abdominal pressure for V-LSO was significantly higher than those of classic LSO and Cybertech®. In the subjective analysis, V-LSO and Cybertech® scored best for comfort. Conclusion: The V-LSO and Cybertech® were more comfortable than the classic LSO, and hence, may have improved compliance with decreased discomfort. V-LSO may be superior to the other LSOs in restricting lumbar movement and increasing intraabdominal pressure.

Application of an Interferometric Biosensor Chip to Biomonitoring an Endocrine Disruptor

  • Kim, Byung-Woo;Lim, Sung-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.9 no.2
    • /
    • pp.118-126
    • /
    • 2004
  • Recombinant E.coli ACV 1003 (recA::lacZ) releasing ${\beta}$-galactosidase by a SOS regulon system, when exposed to DNA-damaging compounds, have been used to effectively monitor endocrine disruptors. Low enzyme activity of less than 10 units/mL, corresponding to a $\mu\textrm{g}$/L(ppb) range of an endocrine disruptor (tributyl tin, bisphenol A. etc.), can be rapidly determined, not by a conventional time-consuming and tedious enzyme assay, but by an alternative interferometric biosensor. Heavily boron-doped porous silicon for application as an interferometer, was fabricated by etching to form a Fabry-Perot fringe pattern, which caused a change in the refractive index of the medium including ${\beta}$-galactosidase. In order to enhance the immobilization of the porous silicon surface, a calyx crown derivative (ProLinker A) was applied, instead of a conventional biomolecular affinity method using biotin. This resulted in a denser linked formation. The change in the effective optical thickness versus ${\beta}$-galactosidase activity, showed a linear increase up to a concentration of 150 unit ${\beta}$-galactosidase/mL, unlike the sigmoidal increase pattern observed with the biotin.