• 제목/요약/키워드: enzymatic profile

검색결과 31건 처리시간 0.023초

An RNA Mapping Strategy to Identify Ribozyme-Accessible Sites on the Catalytic Subunit of Mouse Telomerase

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • 제5권1호
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    • pp.32-35
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    • 2007
  • Telomerase reverse transcriptase (TERT) is an enzymatic ribonucleoprotein that prolongs the replicative life span of cells by maintaining protective structures at the ends of eukaryotic chromosomes. Telomerase activity is highly up-regulated in 85-90% of human cancers, and is predominately regulated by hTERT expression. In contrast, most normal somatic tissues in humans express low or undetectable levels of telomerase activity. This expression profile identifies TERT as a potential anticancer target. By using an RNA mapping strategy based on a trans-splicing ribozyme library, we identified the regions of mouse TERT (mTERT) RNA that were accessible to ribozymes. We found that particularly accessible sites were present downstream of the AUG start codon. This mTERTspecific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors.

섬유소 가수분해반응에 관한 연구(II) - 효소흡착과 가수분해반응에 관여하는 여러인자의 영향 - (Kinetic Studies on Enzymatic Hydrolysis of Cellulose(II) - Evaluation of Several Factors for Enzyme Adsorption and Initial Hydrolysis -)

  • 이용훈;김철
    • KSBB Journal
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    • 제6권2호
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    • pp.167-174
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    • 1991
  • 섬유소 물질을 유용한 자원으로 전환시키기 위하여 섬유소 분해요소를 상요하여 가수분해 반응을 시킬 때 섬유소의 구조적 특성, 섬유소-효소계의 상관성, 가수분해 저해효과, 섬유소 분해효소의 불활성화 등에 의하여 섬유소 가수분해 반응 속도에 상당한 영향을 주는 것으로 알려져 있다. 가수분해 반응이 진행함에 따라 높은 결정성을 가진 섬유소인 Avicel PH 101의 결정화도는 서서히 증가하였으며 무정형이 많은 Solka Floc BW 40은 반응초기에 급격한 증가를 보여주었고 실험적으로 결정성 부분과 무결정성 부분이 동시에 분해됨을 알 수 있었으며, 반응 초기에 급격한 효소의 흡착이 일어나며 반응이 진행함에 따라 서서히 탈착됨을 볼 수 있었다. 반응 생성물인 글루코오스와 셀로바이오스는 초기 가수분해 반응속도에 상당한 저해효과를 주며 또한 생성물의 농도가 증가할수록 효소 흡착량이 감소함을 알 수 있었다. 섬유소 분해요소의 불활성도는 반응시간에 따라 지수 함수적으로 변화함을 실험적으로 알 수 있었다. 셀로바이오스의 초기 가수분해 반응속도는 상대적으로 높은 ${\beta}-glucosidase$의 활성도에 의해 비교적 높은 셀로바이오스의 농도(5-10mg/ ml)에서도 기질의 저해효과가 적음을 볼 수 있었다.

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잎담배 성분중 갈색고분자 물질의 분리정제 및 열분해에 관한 연구 (The Isolation and Pyrolysis of the Brown Pigmented Macromolecule from the Cured Leaf Tobacco)

  • 채쾌;박지창
    • 한국연초학회지
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    • 제2권1호
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    • pp.1-7
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    • 1980
  • 건조잎담배로부터 단백질로 간주되는 고분자 물질의 gel filtration column chromatography(Sephadex G-75), 투석 그리고 흡착 chromatography의 일종인 brushite column chromatography분리를 시도한 결과, 짙은 갈색의 고분자 물질을 얻었다. Sephadex column에 의한 분리 profile를 보면 분자량이 상이한 두종류의 갈색고분자 물질이 존재함을 확인하였고 brushite column의 분리 profile에 의하면 분자의 전자구조가 다른 두 종류의 고분자가 있음이 나타났다. Burley와 Hicks의 경우 단백질 분해효소에 의한 분해효과를 측정해본 결과, 가장 큰 분해효과를 보인 효소는 chymotrypsin으로 Burley에서 16-30%수준까지 분해현상을 나타내 주었으며 Hicks인 경우 38-57%까지 감소현상을 보여 주었다. Pepsin처리효과는 chymotrypsin처리구와 비슷한 수준을 보였으나 trypsin경우는 매우 낮은 분해현상을 나타냈다. 단백질 분해효소로 처리된 시료의 sephadex column분리 profile을 살펴보면, 분자량이 보다 큰 fraction의 경우 peak가 거의 완전하게 사라짐을 관찰할 수 있으나, 작은 분자량Peak는 약간의 감소현상만을 보여 주었다. 투석후의 갈색 고분자물질을 $300^{\circ}C$에서 연소시킨 경우, 연소전에 관찰할 수 없었던 강한 형광성 물질이 생성되었는데 TLC에 의한 분리후, 이 형광성 물질끈 흡수 스펙트라를 측정해 본 결과, 최대흡수 파장이 265. 275nm(benzene용매)로 나타났으며 스펙트럼 끈 모양이 polynuclear aromatic hydrocarbon(PAH) 계열 화합물의 것과 유사함을 보여 주었다.

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Effects of dietary Spirulina on antioxidant status, lipid profile, immune response and performance characteristics of broiler chickens reared under high ambient temperature

  • Mirzaie, Sara;Zirak-Khattab, Fahim;Hosseini, Seyed Abdollah;Donyaei-Darian, Hamid
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권4호
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    • pp.556-563
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    • 2018
  • Objective: Spirulina has been recognized formerly as a filamentous spiral-shaped blue- green algae but more recently as a genus of photosynthetic bacteria (Arthrospira). This microorganism is considered as a rich source of essential nutrients for human and animals. The present study was conducted to determine potential application of Spirulina for heat- exposed broilers. Methods: Two hundred and fifty Cobb 500 chicks with male to female in equal ratio with average initial weight of 615.6 g at 17 days of age were divided into 5 treatments with 5 replicates of 10 chicks. Treatment groups were as follows: positive and negative controls with 0% Spirulina supplement and three Spirulina receiving groups with 5 g/kg (0.5%), 10 g/kg (1%), and 20 g/kg (2%) supplementation. Spirulina receiving groups as well as positive control were exposed to high ambient temperature at $36^{\circ}C$ for 6 h/d from 38 to 44 days of age. Biochemical variables were measured in serum samples at 35, 38, 42, and 45 days of broiler chickens age. Results: The results showed that supplementation of the diet with Spirulina decreased concentration of stress hormone and some serum lipid parameters while enhanced humoral immunity response and elevated antioxidant status whereas it didn't meaningfully affect performance characteristics. Nevertheless, feed conversion ratio was improved numerically but not statistically in broilers fed with 1% Spirulina under high ambient temperature. Conclusion: Overall, the present study suggests that alleviation of adverse impacts due to high ambient temperature at biochemical level including impaired enzymatic antioxidant system, elevated stress hormone and lipid profile can be approached in broiler chickens through supplementation of the diet with Spirulina platensis.

Acetone-Butanol-Ethanol (ABE) Production in Fermentation of Enzymatically Hydrolyzed Cassava Flour by Clostridium beijerinckii BA101 and Solvent Separation

  • Lepiz-Aguilar, Leonardo;Rodriguez-Rodriguez, Carlos E.;Arias, Maria Laura;Lutz, Giselle
    • Journal of Microbiology and Biotechnology
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    • 제23권8호
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    • pp.1092-1098
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    • 2013
  • Cassava constitutes an abundant substrate in tropical regions. The production of butanol in ABE fermentation by Clostridium beijerinckii BA101 using cassava flour (CF) was scaled-up to bioreactor level (5 L). Optimized fermentation conditions were applied; that is, $40^{\circ}C$, 60 g/l CF, and enzymatic pretreatment of the substrate. The batch fermentation profile presented an acidogenic phase for the first 24 h and a solventogenic phase afterwards. An average of 37.01 g/l ABE was produced after 83 h, with a productivity of 0.446 g/l/h. Butanol production was 25.71 g/l with a productivity of 0.310 g/l/h, high or similar to analogous batch processes described for other substrates. Solvent separation by different combinations of fractioned and azeotropic distillation and liquid-liquid separation were assessed to evaluate energetic and economic costs in downstream processing. Results suggest that the use of cassava as a substrate in ABE fermentation could be a cost-effective way of producing butanol in tropical regions.

Properties of Penicillin Amidohydrolase Immobilized on Nylon Fiber

  • B. L. Seng;Iw-Han Cho;J. S. Rhee;Dewey D. Y. Ryu
    • Bulletin of the Korean Chemical Society
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    • 제1권1호
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    • pp.10-17
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    • 1980
  • Penicillin amidohydrolase was partially purified from the fermented broth of Bacillus megaterium, and was immobilized on nylon fiber. The surface area of nylon fiber was increased by roughening it with fine sand and activated by acid treatment. The free amino groups on the nylon fiber exposed by such treatment were then utilized to immobilize the penicillin amidase. Enzymatic properties of penicillin amidohydrolase immobilized on the nylon fiber by covalent bonding and cross linking with glutaraldehyde were studied and compared with those of soluble enzyme. The optimal pH and temperature profile of immobilized enzyme showed only slightly broader peaks, and the values of kinetic constants, $K_m$, $K_{ia}$, and $K_{ip}$, of the immobilized enzyme are only slightly greater than those of the soluble enzyme. These results suggest that the mass transfer effect on the reaction rate for the penicillin amidase immobilized on nylon fiber is not so significant as the enzyme immobilized on some other support material like bentonite. The experimental results of batch reaction agreed well with the results of computer simulation for both the immobilized and soluble enzyme systems, confirming the validity of the rate equation derived which was based on the combined double inhibition by two reaction products.

Gastroprotective effect of cirsilineol against hydrochloric acid/ethanol-induced gastric ulcer in rats

  • Gong, Guojin;Zhao, Rigetu;Zhu, Yuhui;Yu, Jinfeng;Wei, Bin;Xu, Yan;Cui, Zhaoxun;Liang, Guoying
    • The Korean Journal of Physiology and Pharmacology
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    • 제25권5호
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    • pp.403-411
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    • 2021
  • This study was designed to evaluate the gastroprotective activity of cirsilineol in hydrochloric acid (HCl)/ethanol-induced gastric ulcer model. Cirsilineol was administered at the doses of 20 and 40 mg/kg in HCl/ethanol-induced rats. The gastroprotective ability was verified by determining the ulcer score, total acidity, hemoglobin, inflammatory cytokines, lipid peroxides, and enzymatic antioxidants superoxide dismutase (SOD) and catalase (CAT) in gastric tissue and serum biochemical analysis. The results showed a favorable increase in the hemoglobin level, antioxidant enzymes (SOD and CAT), restored electrochemical balance (carbon dioxide & anion gap) while a noticeable decrease in ulcer index, total acidity, lipid peroxides, inflammatory cytokines (interleukin-1 beta [IL-1β], IL-6, and tumor necrosis factor alpha) in rats treated with the cirsilineol. The serum biochemical analysis on liver markers (alkaline phosphatases, alanine aminotransferase, and aspartate aminotransferase), kidney markers (urea, creatinine, albumin, globulin, total protein), and lipid profile (triglyceride, high-density lipoprotein, total cholesterol) were attenuated by cirsilineol treatment in rats. Histopathology showed enhanced gastric protection and preserved the integrity of gastric mucosa upon cirsilineol administration. These results ultimately suggest that cirsilineol has gastroprotective effects that prevent the development of gastric ulcer.

쑥을 이용한 과산화수소 정량 바이오센서의 전기화학적 성질 (Electrochemical properties of the mugwort-embedded biosensor for the determination of hydrogen peroxide)

  • 이범규;박성우;윤길중
    • 분석과학
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    • 제19권1호
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    • pp.58-64
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    • 2006
  • 탄소반죽에 쑥조직을 혼입시켜 과산화수소 정량 바이오센서를 제작하고 그것의 전기화학적 성질을 조사하였다. 다른 생체조직을 이용하여 제작한 센서와 비교할 때, 생촉매 안정성이 뛰어났으며, 보다 큰 감응신호를 보여주었다. 조직의 함량 변화에 따른 신호의 변화는 넓은 범위에 걸쳐 직선성을 보여 주었고, Lineweaver-Burk 도시의 직선성은 전극 감응이 효소 촉매작용에 의하여 조절되고 있음을 보였다. 또 감응전류의 pH 의존성 변화는 센서가 두 종류의 동위효소를 포함하고 있음을 암시하였다.

Cocoa: a functional food that decreases insulin resistance and oxidative damage in young adults with class II obesity

  • Jose Arnold Gonzalez-Garrido;Jose Ruben Garcia-Sanchez;Carlos Javier Lopez-Victorio;Adelma Escobar-Ramirez;Ivonne Maria Olivares-Corichi
    • Nutrition Research and Practice
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    • 제17권2호
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    • pp.228-240
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    • 2023
  • BACKGROUND/OBJECTIVES: Cocoa consumption is associated with health benefits due to its high content of polyphenols. However, the effects of short-term cocoa consumption remain unclear. We aimed to determine the effects generated by cocoa consumption (for 7 days) in young adults in normoweight and class II obesity. SUBJECTS/METHODS: Before-and-after study was carried out in normoweight (NW) (n = 15) and class II obesity (CIIO) (n = 15) young adults. The NW and CIIO participants consumed 25 and 39 g of cocoa, respectively, per day for 7 days. The effect of cocoa consumption was evaluated on the lipid profile, insulin resistance (IR), and inflammation. Oxidative damage was also examined by assessing the biomarkers of oxidative damage in plasma. In addition, recombinant human insulin was incubated with blood obtained from the participants, and the molecular damage to the hormone was analyzed. RESULTS: Cocoa consumption resulted in decreased low-density lipoprotein-cholesterol in both groups (P = 0.04), while the total cholesterol, high-density lipoprotein cholesterol, and triglycerides were maintained at the recommended levels. Initially, IR was detected in the CIIO group (homeostasis model assessment [HOMA] = 4.78 ± 0.4), which is associated with molecular damage to insulin. Interestingly, intervention with cocoa resulted in improved IR (HOMA = 3.14 ± 0.31) (P = 0.0018) as well as molecular damage to insulin. Finally, cocoa consumption significant decreased the arginase activity (P = 0.0249) in the CIIO group; this is a critical enzymatic activity in the inflammatory process associated with obesity. CONCLUSIONS: The short-term consumption of cocoa improves the lipid profile, exerts anti-inflammatory effects, and protects against oxidative damage. Results of this study indicate that cocoa consumption can potentially improve IR and restore a healthy redox status.

Chemoenzymatic Synthesis of Dual-responsive Amphiphilic Block Copolymers and Drug Release Studies

  • Chen, Peng;Li, Ya-Peng;Wang, Shu-Wei;Meng, Xin-Lei;Zhu, Ming;Wang, Jing-Yuan
    • Bulletin of the Korean Chemical Society
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    • 제34권6호
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    • pp.1800-1808
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    • 2013
  • Dual-responsive amphiphilic block copolymers were synthesized by combining enzymatic ring-opening polymerization (eROP) of ${\varepsilon}$-caprolactone (CL) and ATRP of N,N-dimethylamino-2-ethyl methacrylate (DMAEMA). The obtained block copolymers were characterized by gel permeation chromatography (GPC), $^1H$ NMR and FTIR-IR. The critical micelle concentration (CMC) of copolymer was determined by fluorescence spectra, it can be found that with hydrophilic block (PDMAEMA) increasing, CMC value of the polymer sample increased accordingly, and the CMC value was 0.012 mg/mL, 0.025 mg/mL and 0.037 mg/mL for $PCL_{50}$-b-$PDMAEMA_{68}$, $PCL_{50}$-b-$PDMAEMA_{89}$, $PCL_{50}$-b-$PDMAEMA_{112}$, $PCL_{50}$-b-$PDMAEMA_{89}$ was chosen as drug carrier to study in vitro release profile of anti-cancer drug (taxol). The temperature and pH dependence of the values of hydrodynamic diameter (Dh) of micelles, and self-assembly of the resulting block copolymers in water were evaluated by dynamic light scattering (DLS). The result showed that with the temperature increasing and pH decreasing, the Dh decreased. Drug-loaded nanoparticles were fabricated using paclitaxel as model. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) had been explored to study the morphology of the hollow micelles and the nanoparticles, revealing well-dispersed spheres with the average diameters both around 80 nm. In vitro release kinetics of paclitaxel from the nanoparticles was also investigated in different conditions (pH and temperature, etc.), revealing that the drug release was triggered by temperature changes upon the lower critical solution temperature (LCST) at pH 7.4, and at $37^{\circ}C$ by an increase of pH.