• 한국응용과학기술학회지
• /
• 제29권3호
• /
• pp.531-542
• /
• 2012

본 논문은 바이오폴리우레탄의 연구동향에 관한 것이다. 바이오폴리우레탄은 원료 중의 식물유 폴리올과 이소시아네이트의 중부가 고분자이다. 피마자유의 주성분은 히드록시기를 갖는 리시놀산의 트리글리세라이드이다. 이외의 히드록시기가 없는 식물유는 이중결합 위치에서 에폭시화 후 고리열림, 히드로포르밀화 후 수소첨가, 가오존분해 후 수소첨가, 티올-엔 반응으로 히드록시기를 부여한다. 폴리올의 반응성 및 마이크로도메인의 모폴로지 조절을 위한 하이퍼브랜치 폴리올, 일차 알코올 폴리올, 다당류 폴리올이 있다. 의료용의 생분해성 폴리락트산 폴리올, 가수분해 방지용 지방산 다이머 폴리올, 이온성 기를 함유한 수 분산 폴리우레탄용 폴리올이 있다. 바이오폴리올을 이용한 바이오폴리우레탄은 경질 및 연질 폼, 코팅제, 접착제, 실런트, 엘라스토머에 쓰인다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권22호
• /
• pp.9847-9851
• /
• 2014

Background: Multiple myeloma (MM) is a haematological cancer characterized by clonal proliferation of plasma cells.The aim of this study was to investigate the activity of serum paraoxonase-1 (PON1) and arylesterase (ARE) in multiple myeloma with and without free light chain excretion(FLCe-MM and NFLCe-MM); as well as to investigate possible alterations in oxidative stress parameters. Materials and Methods: Total thiol (T.thl), oxidative stress index (OSI), total oxidant status (TOS) and total antioxidant status (TAS) were examined in addition to the PON1 and ARE enzyme activities in twenty one FLCe-MM and nineteen NFLCe-MM subjects. Routine parameters like lipid panel, serum total protein, albumin, creatinine, blood urea nitrogen (BUN), uric acid and hemoglobin levels were compared with the oxidative stress markers. Results: Serum total protein, BUN, creatinin, and uric acid levels were significantly higher (p=0.04, p=0.001, p=0.001 and p=0.0022, respectively), while hemoglobin and albumin levels were significantly lower in FLCe-MM patients (p=0.009 and p=0.04,respectively). PON1 and ARE activities were significantly lower in patients with FLCe-MM compared to those with NFLCe-MM (p=0.001 and p=0.008, respectively). Conclusions: Depending on our results of prognostic markers of MM such as age, hemoglobin, albumin, and creatinine we feel confident to presume FLCe-MM as a subgroup with a worse prognosis. A decrease in PON1 and ARE activities may contribute to the prognosis and may be used as a prognostic tool in MM.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권23호
• /
• pp.10151-10156
• /
• 2015

4-Hydroxynonenal (4-HNE) is a stable end product of lipid peroxidation, which has been shown to play an important role in cell signal transduction, while increasing cell growth and differentiation. 4-HNE could inhibit phosphatase and tensin homolog (PTEN) activity in hepatocytes and increased levels have been found in human invasive breast cancer. Here we report that 4-HNE increased the cell growth of breast cancer cells as revealed by colony formation assay. Moreover, vascular endothelial growth factor (VEGF) expression was elevated, while protein levels of hypoxia inducible factor 1 alpha (HIF-$1{\alpha}$) were up-regulated. Sirtuin-3 (SIRT3), a major mitochondria NAD+-dependent deacetylase, is reported to destabilize HIF-$1{\alpha}$. Here, 4-HNE could inhibit the deacetylase activity of SIRT3 by thiol-specific modification. We further demonstrated that the regulation by 4-HNE of levels of HIF-$1{\alpha}$ and VEGF depends on SIRT3. Consistent with this, 4-HNE could not increase the cell growth in SIRT3 knockdown breast cancer cells. Additionally, 4-HNE promoted angiogenesis and invasion of breast cancer cells in a SIRT3-dependent manner. In conclusion, we propose that 4-HNE promotes growth, invasion and angiogenesis of breast cancer cells through the SIRT3-HIF-$1{\alpha}$-VEGF axis.

• 한국미생물·생명공학회지
• /
• 제14권6호
• /
• pp.441-446
• /
• 1986

Pseudomonas synxantha A3가 생산하는 세포외 guanine deaminase의 몇가지 성질을 검토하였다. 본 효소의 안정 pH는 6.5-7.5 부근이었으며, 열안정성을 검토하기 위하여 각 온도에서 10분간 처리하였을 때 4$0^{\circ}C$까지 안정하였고 그 이상의 온도에서는 서서히 실활되었다. 또한 pH 8.0의 0.2M potassium phosphate 완충액에 본 효소를 보관하였을 때 실온에서 30일간 안정하였고 alcohol 및 acetone은 본 효소의 안정화에 효과가 없었다. 효소활성을 위한 최적온도 및 pH는 각각 5$0^{\circ}C$와 pH 7-8 부근이었다. 본 효소는 1mM의 Hg$^{++}$, Ag$^+$, Li$^+$에 의해 각각 50%이상 저해되었으며, 0.1mM의 Ag$^+$에 의해서도50%이상 저해되었다. Li$^+$에 의해 저해된 본 효소에 EDTA를 가하였을 때 효소의 활성회복에 효과가 있었다. 본 효소의 활성은 1mM의 pentachlorophenol에 의해 50%, p-CMB에 의해 40%정도 저해되었다. p-CMB에 의해 저해된 본 효소에 thiol compound를 가하였을 때에는, 사용된 시약 중 glutathione이 본 효소의 활성회복에 효과적인 것으로 나타났다.

• Journal of Microbiology and Biotechnology
• /
• 제27권6호
• /
• pp.1120-1127
• /
• 2017

Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.

• 미생물학회지
• /
• 제31권3호
• /
• pp.230-236
• /
• 1993

Micrococcus luteus 가 생산하는 isocitrate lyase 의 효소단백질을 ammonium sulfate precipitation, DEAE-cellulose, DEAE-sephacel, 1차 Sephadex G-200, 2 차 Sephadex G-200 column chromatography 과정을 통하여 분리 정제하였으며 정제된 효소의 분자량과 효소학적 특성을 조사하였다. 정제된 isocitrate lyase 는 상기 정제 과정을 통하여 10.2%의 회수율과 38.8 의 정제도를 나타내었으며, 전기영동시 단일밴드를 얻을 수 있었고, SDS-PAGE 에 의해 측정된 분자량을 약 60,000이며 1개의 subunit 로 나타났다. 본 효소의 최적 pH 는 7.5이었으며, 최적 온도는 $40^{\circ}C$ 부근이었고 $45^{\circ}C$까지는 안정하였다. 2가 금속염의 첨가시$ Mg^{2+}$ 를 제외한 다른 금속이온들은 효소활성을 저해하는 것으로 나타났으며 $Mg^{2+}$의 농도는 5 mM 에서 최대활성을 나타내었다. 또한 기질인 DL-isocitrate 의 $K_{m}$ 치는 0.95 mM 로 나타났으며, thiol 화합물의 첨가시 cysteine 은 1mM, glutathione과 $\beta$-mercaptoethanol 은 5 mM 의 농도에서 효소활성이 최대에 달하였다.

• International Journal of Oral Biology
• /
• 제33권3호
• /
• pp.117-123
• /
• 2008

Reactive oxygen species (ROS) are toxic agents that may be involved in various neurodegenerative diseases. Recent studies indicate that ROS can act as modulators of neuronal activity, and are critically involved in persistent pain primarily through spinal mechanisms. In the present study, whole cell patch clamp recordings were carried out to investigate the effects of tert-buthyl hydroperoxide (t-BuOOH), an ROS, on neuronal excitability and the mechanisms underlying changes of membrane excitability. In current clamp condition, application of t-BuOOH caused a reversible membrane depolarization and firing activity in substantia gelatinosa (SG) neurons. When slices were pretreated with phenyl-N-tert-buthylnitrone (PBN) and ascorbate, ROS scavengers, t-BuOOH failed to induce membrane depolarization. However, isoascorbate did not prevent t-BuOOH-induced depolarization, suggesting that the site of ROS action is intracellular. The t-BuOOH-induced depolarization was not blocked by pretreatment with dithiothreitol (DTT), a sulfhydryl-reducing agent. The membrane-impermeant thiol oxidant 5,5-dithiobis 2-nitrobenzoic acid (DTNB) failed to induce membrane depolarization, suggesting that the changes of neuronal excitability by t-BuOOH are not caused by the modification of extrathiol group. The t-BuOOH-induced depolarization was suppressed by the phospholipase C (PLC) blocker U-73122 and inositol triphosphate ($IP_3$) receptor antagonist 2-aminoethoxydiphenylbolate (APB), and after depletion of intracellular $Ca^{2+}$ pool by thapsigargin. These data suggest that ROS generated by peripheral nerve injury can induce central sensitization in spinal cord, and t-BuOOH-induced depolarization may be regulated by intracellular $Ca^{2+}$ store mainly via $PLC-IP_3$ pathway.

• Bulletin of the Korean Chemical Society
• /
• 제28권12호
• /
• pp.2303-2309
• /
• 2007

Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.

• International Journal of Oral Biology
• /
• 제37권4호
• /
• pp.181-188
• /
• 2012

As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 ${\mu}M$. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.

• 한국전기전자재료학회:학술대회논문집
• /
• 한국전기전자재료학회 2004년도 추계학술대회 논문집 Vol.17
• /
• pp.452-455
• /
• 2004

In this paper, investigations of the SAMs(self-assembled monolayers) of a thiol-fuctionalized viologen derivatives, $V_8SH$ and $SH_8V_8SH$, where, V is N,N'-dialkylbipyridinium (i.e. a viologen group), have been carried out by elucidate voltammetry date. The redox reactions are highly reversible and can be cycled many times without significant side reaction, which has been known as a nano-gram order mass detector through resonant frequency change self-assembly process of the viologen has been investigated with $QCM({\Delta}F)$. The assembling process of the $V_8SH$ and $SH_8V_8SH$ monolayers can be finished completely in about 1 hour. The measured frequency shift for $V_8SH$ and $SH_8V_8SH$ were about 351 and 172 Hz, respectively. From these values, we calculated that the mass adsorbed $V_8SH$ and $SH_8V_8SH$ were about 375 and 183 ng. We believe that this mass loss is caused by the simultaneous loss of the anions present within the monolayer for charge compensation of the viologen dications and some solvent.