• Title/Summary/Keyword: Enzyme assays

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In vitro antioxidant property and α-glucosidase and pancreatic lipase inhibiting activities of Jeju camellia mistletoe (Korthalsella japonica (Thunb.) Engl.) extracts (제주 동백나무 겨우살이(Korthalsella japonica (Thunb.) Engl.)의 항산화 및 α-glucosidase와 pancreatic lipase 저해 활성)

  • Park, Eun Mi;Kim, Min Young
    • Journal of Applied Biological Chemistry
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    • v.60 no.3
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    • pp.241-244
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    • 2017
  • The antioxidant activity of various solvent extracts from Jeju camellia mistletoe (Korthalsella japonica (Thunb.) Engl.) was investigated using various in vitro assays as the 1,1-diphenyl-2-picrylhydrazyl radical scavenging, ferrous ion chelating and reducing power assays. Methanol and ethanol extracts showed the most potent antioxidant activity in all assays tested followed by water extract. The inhibitory effect of the Jeju camellia mistletoe extracts on pancreatic lipase and $\acute{a}$-glucosidase was also evaluated and the results showed that methanol and ethanol extracts markedly reduced both enzyme activities. Therefore, the methanol and ethanol extracts of Jeju camellia mistletoe is definitely worthy of further investigation for these beneficial effects on nutraceutical medicine.

Biochemical Study of Recombinant PcrA from Staphylococcus aureus for the Development of Screening Assays

  • Dubaele, Sandy;Martin, Christophe;Bohn, Jacqueline;Chene, Patrick
    • BMB Reports
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    • v.40 no.1
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    • pp.7-14
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    • 2007
  • Helicases are ubiquitous enzymes, which utilize the energy liberated during nucleotide triphosphate hydrolysis to separate double-stranded nucleic acids into single strands. These enzymes are very attractive targets for the development of new antibacterial compounds. The PcrA DNA helicase from Staphylococcus aureus is a good candidate for drug discovery. This enzyme is unique in the genome of S. aureus and essential for this bacterium. Furthermore, it has recently been published that it is possible to identify inhibitors of DNA helicases such as PcrA. In this report, we study the properties of recombinant PcrA from S. aureus purified from Escherichia coli to develop ATPase and helicase assays to screen for inhibitors.

Alpha-Amylase Immobilization on Epoxy Containing Thiol-Ene Photocurable Materials

  • Cakmakci, Emrah;Danis, Ozkan;Demir, Serap;Mulazim, Yusuf;Kahraman, Memet Vezir
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.205-210
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    • 2013
  • Thiol-ene polymerization is a versatile tool for several applications. Here we report the preparation of epoxide groups containing thiol-ene photocurable polymeric support and the covalent immobilization of ${\alpha}$-amylase onto these polymeric materials. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) coupled with SEM was used to explore the chemical composition. The polymeric support and the immobilization of the enzyme were characterized by FTIR analysis. SEM-EDS and FTIR results showed that the enzyme was successfully covalently attached to the polymeric support. The immobilization efficiency and enzyme activity of ${\alpha}$-amylase were examined at various pH (5.0-8.0) and temperature ($30-80^{\circ}C$) values. The storage stability and reusability of immobilized ${\alpha}$-amylase were investigated. The immobilization yield was $276{\pm}1.6$ mg per gram of polymeric support. Enzyme assays demonstrated that the immobilized enzyme exhibited better thermostability than the free one. The storage stability and reusability were improved by the immobilization on this enzyme support. Free enzyme lost its activity completely within 15 days. On the other hand, the immobilized enzyme retained 86.7% of its activity after 30 days. These results confirm that ${\alpha}$-amylase was successfully immobilized and gained a more stable character compared with the free one.

Enzyme Profiles of Alga-Lytic Bacterial Strain AK-13 Related with Elimination of Cyanobacterium Anabaena cylindrica

  • Kim, Jeong-Dong;Han, Myung-Soo
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.184-191
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    • 2004
  • To investigate bacteria with algalytic activities against Anabaena cylindrica when water blooming occurs and to study enzyme profiles associated with alga-lytic activity, various bacterial strains were isolated from surface waters and sediments in eutrophic lakes or reservoirs in Korea. Among 178 isolates, only nine isolates exhibited lytic abilities against A cylindrica on the agar plates, and then the isolate AK-13 was selected as the strongest in lysing the cyanobacterium A. cytindrica. The strain AK-13 was characterized and identified as Sinorhizobium sp. based on fatty acid methyl ether profiles and 16S rDNA sequence. According to the results of the enzyme assays, in the strain An-13 of Sinorhizobium sp., alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase was produced, namely CMCase, laminarinase and protease were highly active. None of glycosidase was produced. Therefore, enzyme systems of Sinorhizobium sp. AK-13 were very complex to degrade cell walls of A. cylindrica. The peptidoglycans of A. cylindrica mat be hydrolyzed and metabolized to a range of easily utilizable monosaccharides or other low molecular weight organic substances by Sinorhizobium sp. AK-13.

Water-extract of Helianthus annuus Seed Exhibits Potent Anti-asthma Activity In Vitro and In Vivo (해바라기씨 추출물의 천식 완화 효과)

  • Heo Jin-Chul;Park Ja-Young;Woo Sang-Uk;Chung Shin-Kyo;Jeong Kyu-Shick;Lee Jin-Man;Ma Jin-Yeol;Lee Sang-Han
    • Food Science and Preservation
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    • v.13 no.4
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    • pp.495-500
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    • 2006
  • In order to investigate whether antioxidant biomaterials inhibits IL-4 and/or IL-13 expression in vitro and in vivo, we carried out antioxidant assays by enzyme or cell-based assays with Helianthus annuus extract. Antioxidant assays include DPPH, FRAP, hydroxyl radical assays. Helianthus annuus extract exhibited SOD scavenging activity, and had different patterns by each solvent extracted reaction. DW extract inhibited oxidative stress by $H_2O_2$ that induced apoptosis. We measured $CD4^+$ cell and IL-/13 cytokine expression in a classical mouse animal model. The result show that Helianthus annuus extract showed strung inhibition of immune response in the lung. These result suggest that Helianthus annuus extract can reduce inflammation induced by n mouse asthma model.

Protective effects of mealworm (Tenebrio molitor) extract on N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced cellular toxicity in SH-SY5Y neuroblastoma cells (SH-SY5Y 인간 신경모세포종 세포에서 MPTP 유발 세포 독성에 대한 거저리(Tenebrio molitor) 추출물의 보호효과)

  • In Ho Jo;Yoo Ji Kim;Seon Tae Kim
    • Journal of The Korean Society of Clinical Toxicology
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    • v.21 no.2
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    • pp.81-91
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    • 2023
  • Purpose: Edible insect extracts have been used as an alternative source for medicinal supplements due to their significant antioxidative and anti-inflammatory activity. Recent studies have reported that anti-microbial peptides from insects have neuroprotective effects on dopamine toxins. The purpose of this study was to investigate the protective functions of mealworm (Tenebrio molitor) extract (MWE) on N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced cellular toxicity in SH-SY5Y neuroblastoma cells. Methods: Cellular toxicity induced by the MPTP toxin and the impact of MWE on cell survival were analyzed using MTT assays. DAPI staining was performed to observe apoptotic phenomena caused by MPTP. Changes in caspase-3 activity and protein expression were observed using enzyme activity assays and western blot assays, respectively. Results: MWE exerted significant antioxidant activity, which was measured by both DPPH and ABTS radical assays, with a dose-dependent relationship. Furthermore, MWE resulted in cellular proliferation in SHSY5Y cells in a dose-dependent manner. Furthermore, MWE pretreatment significantly inhibited MPTP-induced cytotoxicity, with a dose-dependent relationship. The morphological characteristics of apoptosis and increased reactive oxygen species induced by MPTP were also significantly reduced by MWE pretreatment. Conclusion: MWE treatment significantly attenuated MPTP-induced changes in the levels of proteins associated with apoptosis, such as caspase-3 and PARP. These findings suggest that MWE exerts neuroprotective effects on human neuroblastoma SH-SY5Y cells subject to MPTP-induced dopaminergic neurodegeneration.

Synthesis of New Diselenide Compounds as Anti-Inflammatory Agents

  • Shen, Liulan;Shin, Kyung-Min;Lee, Kyung-Tae;Jeong, Jin-Hyun
    • Archives of Pharmacal Research
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    • v.27 no.8
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    • pp.816-819
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    • 2004
  • Many diselenide compounds are used as antioxidants, enzyme inhibitors and cytokine induc-ers. Three new diselenide compounds, bis-(2-hydroxyphenyl) diselenide, bis-(3-hydroxyphe-nyl) diselenide and bis-(4-hydroxyphenyl) diselenide were designed and synthesized as anti-inflammatory agent. All of them were found to have strong in vitro activity in anti-inflammatory assays.

Assay development and HTS on microfluidic Lab-on-a-chip

  • Yang, Eun-Gyeong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2002.07a
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    • pp.73-78
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    • 2002
  • Microfluidic lab-on-a-chip (LOC) systems have enabled a new generation ofassay technologies in chemical and biomedical sciences. Caliper's microfluidic LOC systems contain a network of microscopic channels through which fluids and chemical are moved in order to perform experiments. The main advantages of these continuous-flow devices are integration and automation of multiple steps in complex analytical procedures to improve the reproducibility of the results, and eliminated the manual labor, time and pipetting errors involved in analyses. The present talk is devoted to give a brief introduction of microfluidic basics and to present in applying continuous-flow microchips to drug screening with model enzyme assays.

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Design of Smart Polymer Constructs for Use in Microfluidic Diagnostic Assays

  • Hoffman Allan S.
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.42-43
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    • 2006
  • We have prepared random, block and graft copolymers with single or dual sensitivities to various stimuli. We have conjugated these polymers to proteins at random lysine sites or at specific sites designed into the protein by genetic engineering. We are also grafting the smart polymers to the surfaces of nanobeads. We are applying these smart conjugates and smart nanobeads in microfluidic devices for various applications, including diagnostics, affinity separations and enzyme bioprocesses. In this talk I will update our work with these interesting hybrid systems.

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