• Title/Summary/Keyword: enzymatic activity assay

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Effects of Browning Reaction Products on DNA Damage (효소적 갈변 생성물의 DNA 손상에 대한 효과)

  • Lee, Ji-Eun;Kim, An-Keun
    • Korean Journal of Pharmacognosy
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    • v.31 no.2
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    • pp.240-244
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    • 2000
  • Antimutagenicity profiles of the enzymatic browning reaction products(EBRP) were investigated. The rec-assay with Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$ was carried out using their spores. The biological activities were evaluated for seven different enzymatic browning reaction products, which resulted from the reactions of seven polyphenols with polyphenol oxidase isolated from Ginkgo biloba leaves. In the spore $rec^-$ assay, most of the polyphenolic compounds tested were positive, whereas their enzymatic browning reaction products were tested negative. The mutagenicity of enzymic browning mixtures of the polyphenols and the enzymes obtained from Ginkgo biloba leaves showed negative results in the mutagenicity test using Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$. In the case where polyphenol oxidase inhibitors were added in the enzymatic reaction mixtures with polyphenols, the polyphenols showed mutagenic effect in the spore $rec^-$ assay. This suggests that the activity of polyphenol oxidase is decreased.

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Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols (재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi;Hong, Eun-Hee;Omura, Hirohisa
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.212-219
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    • 1987
  • The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

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The Inhibitory Effect of Polyphenol Oxidase on Polyphenol-Induced DNA Damage of Bacillus subtilis (고초균에서 폴리페놀로 유도된 DNA 손상에 대한 폴리페놀산화효소의 억제효과)

  • Kim An Keun;Kim Yoo Kyung;Kang Young-Sook
    • YAKHAK HOEJI
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    • v.49 no.4
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    • pp.330-334
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    • 2005
  • Antimutagenic activity of the enzymatic browning reaction products (EBRPs) was investigated by using the spore rec-assay with Bacillus subtilis strains H17 $(rec^+)\;and\;M45 (rec^-)$. The EBRPs tested were prepared from the reactions of five different kinds of polyphenols with polyphenol oxidase isolated from the leaves Perilla frutescens. In the spore rec-assay, most of the polyphenolic compounds tested showed positive, whereas only their tested compound showed negative respectively. In addition of polyphenol oxidase inhibitors such as cysteine, glutathione and ascorbic acid to the reaction mixtures consisted with the polyphenol oxidase and polyphenols, the mutagenic effects were increased in the spore recassay. These results show that the activity of polyphenol oxidase may play an important role in the reduction of mutagenicity of polyphenols.

In vitro Anti-obesity Effect of 4-hydroxybenzyl Alcohol from Cudrania tricuspidata

  • Choi, Jun-Hui;Park, Se-Eun;Kim, Myung-Kon;Lee, Hyo-Jeong;Seo, Kyoungsun;Kim, Seung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.81-81
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    • 2018
  • The present study was investigated on in vitro anti-obesity effect of 4-hydroxybenzyl alcohol from Cudrania tricuspidata. We isolated various compounds from Cudrania tricuspidata. Among these compounds, anti-obesity effects of 4-hydroxybenzyl alcohol was examined by lipase activity assay, cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase type IV (PDE4) activity assay, and citrate synthase activity assay. 4-hydroxybenzyl alcohol and Cudrania tricuspidata extracts inhibited the enzymatic activities of lipase, PDE4, and citrate synthase. Lipase is known to mediate the hydrolysis of triacylglycerol in adipose tissue and cholesterol esters in other tissue or cells. Also, PDE4 hydrolyses cAMP, a crucial secondary messenger for in metabolic pathways including glucose and lipid metabolism, lipolysis, and thermogenic function. 4-hydroxybenzyl alcohol and Cudrania tricuspidata extracts induced the inhibitory effect against each enzymatic activity on several specific substrates as observed by detection at 405 or 412 nm. These findings might be attributable to the inhibition of adipogenesis, and partial prevention of obesity. In conclusion, these results show that 4-hydroxybenzyl alcohol and Cudrania tricuspidata may be a critical candidate as a natural anti-obesity source.

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Interactions between Hyaluronic Acid, Lysozyme, Peroxidase, and Glucose Oxidase in Enzymatic Activities at Low pH

  • Kim, Bum-Soo;Kim, Yoon-Young;Chang, Ji-Youn;Kho, Hong-Seop
    • Journal of Oral Medicine and Pain
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    • v.39 no.4
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    • pp.127-132
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    • 2014
  • Purpose: Many substances in saliva or oral health care products interact with each other. The aim of this study was to investigate interactions between hyaluronic acid (HA), lysozyme, peroxidase, and glucose oxidase (GO) in enzymatic activities at low pH levels. Methods: HA (0.5 mg/mL), hen egg-white lysozyme (HEWL, $30{\mu}g/mL$), bovine lactoperoxidase (bLPO, $25{\mu}g/mL$), and GO ($50{\mu}g/mL$) were used. The influences of HA, bLPO, and GO on HEWL activity were determined by measuring the turbidity of a Micrococcus lysodeikticus suspension. The influences of HA and HEWL on bLPO activity were determined by the NbsSCN assay, measuring the rate of oxidation of 5-thio-2-nitrobenzoic acid (Nbs) to 5,5'-dithiobis(2-nitrobenzoic acid) $(Nbs)_2$. The influences of HA and HEWL on GO activity were determined by measuring oxidized o-dianisidine production. All experiments were performed at pH 4, 5, and 6. Results: HA and GO did not affect the enzymatic activity of HEWL at pH 4, 5, and 6. bLPO enhanced the enzymatic activity of HEWL at pH 5 (p<0.05) and pH 6 (p<0.05) significantly. The enzymatic activity of bLPO was not affected by HA and HEWL at pH 4, 5, and 6. HA and HEWL did not affect the enzymatic activity of the GO at pH 4, 5, and 6. Conclusions: Peroxidase enhances lysozyme activity at low pH, otherwise there were no significant interactions in enzymatic activities between HA, lysozyme, peroxidase, and GO at low pH levels.

Desmutagenicity of the Enzymatic Browning Reaction Products Which Obtained from Prunus salicina (yellow) Enzyme and Polyphenol Compounds (재래종 황색자두효소 갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.71-76
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    • 1987
  • The mutagenicity and desmutagenicity on enzymatic browning reaction products which obtained from prunes salicina (yellow) enzyme and polyphenol compounds were carried out. In the rec-assay on Bacillus subtilis strains H17 and M45, the enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihydroxytoluene and catechol of $10^{-2}M$ did not showed mutagenicity. In the effects of various metal ions on the rec-assay, the enzymatic browning reaction products of pyrogallol showed mutagenic activity by $Fe^{3+},\;Mn^{2+},\;Zn^{2+},\;Ni^{2+}$ and $Al^{3+}$. In the enzymatic browning reaction products of hydroxyhydroquinone, $Cu^{2+},\;Mn^{2+}$ and $Pb^{2+}$ were effected in mutagenic action and the enzymatic browning reaction products of catechol was effected in mutagenic action by $Mn^{2+}$. In the DNA-breaking action of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihyroxytoluene and catechol did not show, DNA-breaking action. In the effects of various metal ions on the DNA-breaking action of enzymatic browning reaction products, $Cu^{2+}$ showed DNA-breaking action. In the mutagenicity test on Sal. typhimurium strains TA98 and TA 100 with S-9 mix, 4 kinds of browned substances did sot shove muragenicity, all the browned substances showed strong desmutagenic activity in the presence of benzo $({\alpha})-pyrene$ with S-9 mix.

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Localization of Germin Genes and Their Products in Developing Wheat Coleoptiles

  • Caliskan, Mahmut;Ozcan, Birgul;Turan, Cemal;Cuming, Andrew C.
    • BMB Reports
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    • v.37 no.3
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    • pp.339-342
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    • 2004
  • Germination is a process which characterized with nescient synthesis of genes. Among the genes synthesized during the germination of wheat embryos, germin genes, proteins and their enzymatic activity were defined. Germin is a water soluble homopentameric glycoprotein which is remarkable resistant to degradation by a broad range of proteases including pepsin. Germin proteins found to have strong oxalate oxidase activity which produces hydrogen peroxide by degrading oxalic acid. The current study, aimed to localize the germin genes, proteins and enzymatic activities in developing coleoptiles which is a rapidly growing protective tissue of leaf primordium and shoot apex. Non-radioactively abeled germin riboprobes were employed to localize germin mRNAs in situ. FITC (Fluorescein isothiocyanate) and alkaline phosphatase linked anti-germin antibodies were used to localize germin proteins under the fluorescence and light microscopy and finally germin enzymatic activity was localized by using appropriate enzyme assay. The results revealed that in coleoptiles germin genes, proteins and their enzymatic activity were predominantly associated with the cells of epidermis and vascular bundle sheath cells.

Biological Activity and Inhibition of Non-Enzymatic Glycation by Methanolic Extract of Rosa davurica Pall. Roots

  • Hu, Weicheng;Han, Woong;Jiang, Yunyao;Wang, Myeong-Hyeon;Lee, Young-Mee
    • Preventive Nutrition and Food Science
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    • v.16 no.3
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    • pp.242-247
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    • 2011
  • The methanolic extract of Rosa davurica Pall. roots exhibited strong antioxidant activity in a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay and was found to be a dose-dependent inhibitor of non-enzymatic formation of advanced glycation end products (AGEs), which are relevant to diabetes complications. HPLC-diode array detector (DAD) analysis of the R. davurica Pall. root extract led to the identification of four compounds: hydrocaffeic acid, catechin, epicatechin, and ellagic acid. Catechin was present in the largest amount and exhibited high antiglycation activity. A CYP3A4 assay was used to investigate potential interactions between drugs and the extract, and results suggest that the R. davurica Pall. root extract had moderate potential for interfering with drug metabolism. The R. davurica Pall. extract did not display anti-inflammatory activity on the level of that for tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in a lipopolysaccharide (LPS)-stimulated macrophage assay; however, the extract did exhibit low to moderate immunostimulatory activity in a pro-inflammatory macrophage assay. Therefore, we conclude that R. davurica Pall. root is a promising anti-AGE agent with low to moderate risks of associated inflammation or drug interaction.

Modification of the characteristics of silkworm powder by treatment with alkaline protease

  • Kim, Sung-Kuk;Jo, You-Young;Lee, Kwang-Gill;Kim, Hyun-Bok;Kim, Yong Soon;Ju, Wan-Taek;Jung, Da-Eun;Kweon, HaeYong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.1
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    • pp.30-33
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    • 2015
  • Enzymatic modification of proteins is often used to increase the biological activity of materials. Silkworm powder has been investigated as a functional food resource, but no study has been performed on its modification by commercial food enzyme. Therefore, this study aimed to determine the feasibility of such modification of silkworm powder by alkaline protease. The activity of the enzyme was confirmed using an azocasein assay. Subsequently the silkworm powder was hydrolyzed by enzymatic treatment. UV visible spectrometry showed that the supernatant of silkworm powder subjected to enzymatic treatment had a stronger absorption band than the untreated powder. SDS-PAGE electrophoresis showed that the molecular weight of silkworm powder decreased on enzymatic treatment. Thus the results indicate that commercial enzymes might be used to modify the characteristics of silkworm powder.

Antioxidant Activities and Whitening Effect from Lindera obtusiloba BL. Extract (생강나무 추출물의 항산화 활성과 미백효과)

  • Bang, Chae-Young;Won, Eun-Kyung;Park, Kuen-Woo;Lee, Gwang-Won;Choung, Se-Young
    • YAKHAK HOEJI
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    • v.52 no.5
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    • pp.355-360
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    • 2008
  • In this study we investigated antioxidant activity of against several free radicals and skin whitening effect of 70% ethanol extract (leaf extracts and branch/stem mixed) of Lindera obtusiloba BL. Antioxidant activity was assessed by DPPH, superoxide radical and hydroxyl radical assays. The Lindera obtusiloba BL. extract had antioxidant activity dose dependently with an ${IC}_{50}$ value of 243.14 and 181.10 ${\mu}g$/ml for DPPH, 165.77 and >1500 ${\mu}g$/ml for non-enzymatic system of superoxide radical assay, 35.47 and >100 ${\mu}g$/ml for enzymatic system of superoxide radical assay, 1.21 mg/ml for hydroxyl radical assay. In addition we tested tyrosinase inhibition activity and melanin contents on B16 melanoma F10. B16 melanoma cell was treated by such sample as 1, 5, 10 and 50 ${\mu}g$/ml for 72 hr and tyrosinase inhibition was tested. Melanogenesis was inhibited to 22% at the dose of 50 ${\mu}g$/ml and tyrosinase was inhibited to 45.2% at the same dose. In conclusion Lindera obtusiloba BL had potent antioxidant activity and inhibitory activity of tyrosinase and melanin formation. It could be developed as the health functional food and functional cosmetic resources.