• Title/Summary/Keyword: soluble stability

Search Result 413, Processing Time 0.027 seconds

Purification of Caudal-Related Homeodomain Transcription Factor and Its Binding Characterization

  • Jeong, Mi-Suk;Hwang, Eun-Young;Kim, Hyun-Tae;Yoo, Mi-Ae;Jang, Se-Bok
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.12
    • /
    • pp.1557-1564
    • /
    • 2009
  • Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

Gene Cloning, Expression, and Characterization of a New Carboxylesterase from Serratia sp. SES-01: Comparison with Escherichia coli BioHe Enzyme

  • Kwon, Min-A;Kim, Hyun-Suk;Oh, Joon-Young;Song, Bong-Keun;Song, Jae-Kwang
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.2
    • /
    • pp.147-154
    • /
    • 2009
  • The carboxylesterase-encoding gene(bioHs) of a newly isolated strain, Serratia sp. SES-01, was cloned from the genomic DNA library by detecting formation of transparent halo around the colony on LB-tributyrin agar plates. The amino acid sequence of BioHs was highly similar to the members of the BioH enzyme family involved in the biotin biosynthetic pathway; it showed the highest similarity(91%) with that of Serratia proteamaculans. To compare BioHs with other BioH enzymes, the relatively well-known bioHe gene of E. coli was cloned with PCR. After we achieved high-level expression of soluble BioHs and BioHe through the exploration of different culture conditions, the purified BioHs and BioHe enzymes were characterized in terms of specificity, activity, and stability. BioHe was generally more robust to a change in temperature and pH and an addition of organic solvents than BioHs. The two enzymes exhibited a strong preference for carboxylesterase rather than for thioesterase and were optimal at relatively low temperatures($20-40^{\circ}C$) and alkaline pHs(7.5-9.0). The results in this study strongly suggested that both the BioHs and BioHe enzymes would be potential candidates for use as a carboxylesterase in many industrial applications.

Effect of $Li_2O$ and $Na_2O$ addition on the microstructure and humidity sensitivity of $V_ 2O_5$-doped $TiO_2$ ($V_2O_5$를 도판트한 $TiO_2$의 미세구조와 감습에 미치는 알카리 옥사이드$ M_2O(M=Li,Na)$의 영향)

  • 신용덕
    • Electrical & Electronic Materials
    • /
    • v.9 no.6
    • /
    • pp.605-615
    • /
    • 1996
  • In this paper, the effect of alkaline oxide addition such as Li$_{2}$O and Na$_{2}$O on the microstructure and humidity sensitivity of V$_{2}$O$_{5}$(2 mol%)-doped TiO$_{2}$(98 mol%) was investigated as a function of amount (0, 1, 2, 5, 10 mol%) of Li$_{2}$O and Na$_{2}$O additives. The pores in the alkaline free sample were distributed mostly in the range between 0.16 and 1.0.mu.m in diameter and its porosity was 23.29%. Li$_{2}$O caused grain overgrowth and reduced the porosity with a narrow distribution of the pore size, leading to poor humidity sensitivity. Na$_{2}$O helped to enlarge the distribution of the pore size through the formation of small soluble phases. The pore sizes of the sample containing Na$_{2}$O 2mol% were distributed mostly in the range between 1.0 and 2.5.mu.m in diameter and its porosity and intrusion volume of mercury were 31.13 % and 0.1155 mL/g respectively, which consequently improved the humidity sensing characteristics such as the sensitivity and temperature-stability. Especially, the addition of 2mol% of Na$_{2}$O improved the humidity-sensing characteristics such as sensitivity and linearity in the whole range between 30 and 90 %RH (Percentage Relative Humidity)y)

  • PDF

Anti-diabetic Effects of CCCA, CMESS, and Cordycepin from Cordyceps militaris and the Immune Responses in Streptozotocin-induced Diabetic Mice

  • Yun, Yun-Ha;Han, Shin-Ha;Lee, Seung-Jeong;Ko, Sung-Kwon;Lee, Chong-Kil;Ha, Nam-Joo;Kim, Kyung-Jae
    • Natural Product Sciences
    • /
    • v.9 no.4
    • /
    • pp.291-298
    • /
    • 2003
  • Anti-diabetic effect of various fractions of Cordyceps militaris (CM), CCCA (crude cordycepin containing adenosine), CMESS (ethanol soluble supernatant), and cordycepin were evaluated in streptozotocin (STZ) induced diabetic mice, CMESS showed potent inhibitory activity of 34.7% in starch-loaded mice (2 g/kg) while acarbose as a positive standard exhibited 37.8% of inhibition rate. After 3 days administration (50 mg/kg), cordycepin (0.2 mg/kg), and acarbose (10 mg/kg) dramatically reduced blood glucose level (inhibition ratio: 46.9%, 48.4% and 37.5% respectively). CCCA that has high contents of cordycepin (0.656 mg/4 mg) did not influence on reducing blood glucose level. The proliferation of splenocytes and peritoneal macrophages derived from STZ-induced diabetic mice administered samples were evaluated out by addition of mitogens to see the stability of the usage of these herbal medicines. Proliferation of T-lymphocyte was significantly decreased; while NO production was increased more than two fold to STZ control in the cordycepin-administered group. Changes of serum enzyme levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were also evaluated. Cordycepin administered group was appeared to acarbose. We conclude that CMESS and cordycepin may be useful tools in the control of blood glucose level in diabetes and promising new drug as an anti-hyperglycemic agent without defects of immune responses and other side effects.

Spectrophotometric Study of Copper Complex of N-Benzylisonitrosoacetylacetone Imine (N-Benzylisonitrosoacetylacetone Imine의 구리착물에 대한 분광광도법적 연구)

  • Byung Kyo Lee;Dae Sub O;Heung Rak Lee
    • Journal of the Korean Chemical Society
    • /
    • v.30 no.2
    • /
    • pp.201-206
    • /
    • 1986
  • A new analytical reagent N-benzylisonitrosoacetylacetone imine (H-IAA-N-Bz) has been synthesized and identified its structure by IR, NMR and mass spectra. H-IAA-N-Bz forms a copper chloroform-soluble complex in a basic aqueous solution (pH = 7.0∼10.0). The other optimum conditions for the spectrophotometric study of the copper complex have been determined at 420nm. Beer's law is obeyed below the concentration of 64$\mu$g of copper per 10ml of chloroform. The composition of the copper complex has been found to be $Cu(IAA-N-Bz)_2$ and the over-all stability constant is calculated to be $8.55 {\times} 10^6$. The molar absorption coefficient, $\varepsilon$ of the $Cu-(IAA-N-Bz)_2 $complex is 3500l/$cm{\cdot}mol$.

  • PDF

Hydrophobic and Hydrophilic PDMS Sponges Prepared Through Physicochemical Treatments (물리화학적 처리에 따른 PDMS 특성 조절)

  • Nam, Kyungmok;Park, Sungmin;Kim, Jonghun;Yoon, Sang-Hee
    • Transactions of the Korean Society of Mechanical Engineers A
    • /
    • v.40 no.8
    • /
    • pp.737-742
    • /
    • 2016
  • An elastomer (especially PDMS) sponge is considered to be a promising selective absorber in cleaning up oil spills. The performance of a PDMS sponge in capturing and separating oil from (sea) water depends on several parameters such as surface roughness, physicochemical treatments, and hydrostatic stability. Here, we first present a method of fabricating the PDMS sponges having numerous micro-sized pores that act as absorption and storage spaces for the target material, and then we report an experimental effort undertaken to control the surface physicochemistry (i.e., hydrophobicity or hydrophilicity) of the PDMS sponges by adjusting the size of the pores and the concentration of the surfactant (i.e., silwet L-77). From the experimental results, we develop an in-depth understanding of the mechanism for controlling the surface physicochemistry of PDMS using water-soluble micro-sized particles and a surfactant. The surface energy and absorbing behavior of the PDMS sponges are also extensively discussed.

Immobilization of Cyclodextrin Glucanotrasferase on Amberline IRA-900 for Biosynthesis of Transglycosylated Xylitol

  • Kim, Pan-Soo;Shin, Hyun-Dong;Park, Joong-Kon;Lee, Young-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.3
    • /
    • pp.174-180
    • /
    • 2000
  • Cyclodextrin glucanotransferase (CGTasa) from Thermoanaerobacter sp. was adsorbed on the ion exchange resin Amberlite IRA-900. The optimum conditions for the immobilization of the CGTase were pH6.0 and 600 U CGTase/g resin, and the maximum yield of immobilization was around 63% on the basis of amount ratio of the adsorbed enzyme to intial amount in the solution. Immobilixation of CGTase shifted the optimum temperature for the enzyme to peoduce transglycosylated xylitol from 7$0^{\circ}C$ to 9$0^{\circ}C$ and improved the thermal stability of immobilized CGTase, especially after the addition of soluble starch and calcium ions. Transglycosylated xylitol was continuoncly produced using immobilized CGTase in the column type packed bed reactor, and the operating conditions for maximum yield were 10%(w/v) dextrin (13 of the dextrose equivalent) as the glycosyl donor, 10%(w/v) dextrin (13 of the dextrose equivalent) as the glycosyl donor, 10%(w/v) xylitor as the glycosyl acceptor, 20mL/h of medium fiow rate, and 6$0^{\circ}C$. The maximum yield of transglycosylated xylitol and productivity were 25% and 7.82 g.L-1.h-1, respectively. The half-life of the immobilized CGTase in a column type packed bed reactor was longer than 30 days.

  • PDF

Petrological Characteristics and Deterioration Aspect of the Goryeong Yangjeondong Petroglyph (고령 양전동암각화의 암석학적 특징과 훼손양상 분석)

  • Choi, Gi-Ju;Lee, Sang-Hun
    • Journal of Conservation Science
    • /
    • v.25 no.2
    • /
    • pp.131-145
    • /
    • 2009
  • Goryeong Yangjeondong Petroglyph on the joint face of the bedrock (6m in width, 3m in height) composed of the green sandstone has circles and shield pattern sculptures. The rock is mainly composed of quartz, feldspar, chlorite, illite and calcite with the clay minerals. These clay minerals occur as pore lining, replacement of component and pore fillings. Investigated the weathering aspects including the connection between soluble salt and rock weathering and the stability of the bedrock with petroglyphs. The Major deterioration aspect are grain peel-off, surface exfoliation, cracks and damage. And the bedrock where the joints and cracks advance is interpreted with the fact that has the possibility of the falling rock. For conservation, adhesive need to cover crack and damage and consolidation is necessary in the surface which becomes weak.

  • PDF

Glycosylation Enhances the Physicochemical Properties of Caffeic Acid Phenethyl Ester

  • Moon, Keum-Ok;Park, Soyoon;Joo, Myungsoo;Ha, Ki-Tae;Baek, Nam-In;Park, Cheon-Seok;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
    • /
    • v.27 no.11
    • /
    • pp.1916-1924
    • /
    • 2017
  • In this study, we synthesized a glycosylated derivative of caffeic acid phenethyl ester (CAPE) using the amylosucrase from Deinococcus geothermalis with sucrose as a substrate and examined its solubility, chemical stability, and anti-inflammatory activity. Nuclear magnetic resonance spectroscopy showed that the resulting glycosylated CAPE (G-CAPE) was the new compound caffeic acid phenethyl ester-4-O-${\alpha}-{\small{D}}$-glucopyranoside. G-CAPE was 770 times more soluble than CAPE and highly stable in Dulbecco's modified Eagle's medium and buffered solutions, as estimated by its half-life. The glycosylation of CAPE did not significantly affect its anti-inflammatory activity, which was assessed by examining lipopolysaccharide-induced nitric oxide production and using a nuclear factor erythroid 2-related factor 2 reporter assay. Furthermore, a cellular uptake experiment using high-performance liquid chromatography analysis of the cell-free extracts of RAW 264.7 cells demonstrated that G-CAPE was gradually converted to CAPE within the cells. These results demonstrate that the glycosylation of CAPE increases its bioavailability by helping to protect this vital molecule from chemical or enzymatic oxidation, indicating that G-CAPE is a promising candidate for prodrug therapy.

Preparation of Organic-inorganic Hybrid PES Membranes using Fe(II) Clathrochelate (Fe(II) clathrochelate을 이용한 유.무기 PES 복합막의 제조)

  • Jung, Bo Ram;Son, Yeji;Lee, Yong Taek;Kim, Nowon
    • Membrane Journal
    • /
    • v.23 no.1
    • /
    • pp.80-91
    • /
    • 2013
  • Metal-templated condensation of cyclohexanedione dioxime and phenylboronic acid in the presence of Fe(II) sulfate heptahydrate proceeds cleanly in methanol to furnish the Fe(II) clathrochelate. An organic/inorganic hybrid membranes composed of Fe(II) clathrochelate and polyethersulfone was prepared by using phase inversion method. For membrane preparation, the Fe(II) clathrochelate was highly soluble (3~5 g/L) in DMF, NMP, and DMAc, which meets the requirements for the solubility of metal complexes in polar aprotic solvent used in membrane preparation. It was stable even in the presence of strong acids, such as trifluorosactic acid (pKa = 0.3). It was characterized by UV-vis spectroscopy, and their stability in solution phase studied in the presence of (i) strong acids or (ii) competing chelates. Organic/inorganic hybrid membranes were prepared with polyethersulfone, polyvinylpyrrolidone, p-toluenesulfonic acid, Fe(II) clathrochelate and DMF by using nonsolvent induced phase inversion method. The addition of Fe(II) clathrochelate leads increase of surface pore density, mean pore size and flux. We can obtain highly asymmetric membranes by addition of Fe(II) clathrochelate.