• 제목/요약/키워드: enzymatic activity assay

검색결과 145건 처리시간 0.028초

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

  • 이지은;김안근
    • 생약학회지
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    • 제31권2호
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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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재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용 (Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols)

  • 함승시;홍은희;대촌호구
    • 한국식품과학회지
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    • 제19권3호
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    • pp.212-219
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    • 1987
  • 재래종 적색 자두 (Prunus salicina)에서 효소를 추출하여 4종류의 polyphenol화합물과 반응시켜 얻어진 갈변반응 생성물에 대하여 Bacillus subtilis H17과 M45를 이용한 rec-assay와 Salmonella typhimurium TA98과 TA100 두 균주를 이용한 Ames test, 그리고 calfcthymus DNA를 이용하는 DNA절단시험을 이용하여 돌연변이원성과 돌연변이 억제작용을 조사하였다. 포자 rec-assay 에서는 pyrogallol, hydroxyhydroquinone, 3,4-dihydroxytoluene, chlorogenic acid 의 갈변반응 생성물은 모두 DNA손상능력이 없었으며 8가지 금속이온 중 ${Zn}^{2+}$${Ni}^{2+}$의 첨가로 고초균 DNA손상에 약한 영향을 나타내었다. DNA절단시험 결과 4종류 갈변반응 생성물 모두 DNA절단작용이 없었으며 금속이온의 영향에 있어서는 pyrogallol 갈변반응 생성물이 ${Cu}^{2+}$의 영향을 받아 ${Cu}^{2+}$의 농도가 증가함에 따라 강한 절단작용을 나타내었으며 3,4-dihydroxytoluene 과 hydroxyhydroquinone갈변반응 생성물은 금속이온의 영향을 전혀 받지 않았다. 또한 chlorogenic acid갈변반응 생성물은 DNA 절단을 억제하는 효과를 나타내었다. Ames test에서는 4가지 갈변반응 생성물 모두 변이원성은 없었으며 benzo$[{\alpha}]$pyrene을 사용한 변이원성 억제작용 실험결과 benzo$[{\alpha}]$pyrene의 활성을 강하게 억제하는 것으로 나타났다.

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

  • 김안근;김유경;강영숙
    • 약학회지
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    • 제49권4호
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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
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 춘계학술발표회
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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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    • 제39권4호
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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.

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

  • 함승시
    • Applied Biological Chemistry
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    • 제30권1호
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    • pp.71-76
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    • 1987
  • 재래종 황색계자두 효소와 4종류의 polyphenol 화합물과 반응시켜 얻어진 효소적 갈변반응 생성물에 대하여 변이원성 및 변이원 억제작용에 관한 실험결과 B. subtilis H17과 M45 두 균주를 이용한 rec-assay에서 4종류의 갈변반응 생성물 모두 변이원성은 없었다. rec-assay에서 금속이온의 영향을 실험한 결과 pyrogallol의 갈변반응 생성물의 경우 $Fe^{3+},\;Mn^{2+},\;Zn^{2+},\;Ni^{2+}\;Al^{2+}$에서 약한 영향을 받았으며 hydroxyhydroquinone의 갈변반응 생성물에서는 $Cu^{2+},\;Pb^{2+}$, 그리고 catechol의 갈변반응 생성물에서는 $Mn^{2+}$ 공존하에서 약한 영향을 받았다. DNA절단실험 결과 갈변반응 생성물 자체에 의해서는 4종류의 시료 모두 DNA절단능력은 없었다. 금속이온의 영향에서는 $Fe^{2+}$의 경우 그 자체에 의해 절단되었으며 $Cu^{2+}$은 DNA 절단을 촉진하는 것으로 나타났다. S-9 mix를 첨가한 돌연변이 유발실험에서는 4종류의 갈변반응 생성물 모두 농도증가에도 변이원성은 없었으며 변이원 물질인 $benzo\;{\alpha}\;pyrene$에 대한 돌연변이원 억제반응에서는 4종류의 갈변반응 생성물 모두 농도증가에 따라 대조구에 비해 강한 억제작용을 나타냈다. 따라서 재래종 황색계 자두로부터 추출한 polyphenol oxidase와 4종류의 polyphenol 화합물과 반응시켜 얻어진 효소적 갈변반응 생성물의 들연변이원성 실험결과 갈변반응 생성물자체에 의한 돌연변이원성은 없었으며 금속이온의 영향도 받지 않는 것으로 나타났으나 강력한 발암물질인 $benzo\;{\alpha}\;pyrene$을 사용한 돌연변이 억제실험에서는 4종류의 갈변반응 생성물 모두 강한 돌연변이 억제작용이 있음을 알 수 있었다.

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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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    • 제37권3호
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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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    • 제16권3호
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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.

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

  • 방채영;원은경;박권우;이광원;정세영
    • 약학회지
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    • 제52권5호
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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.

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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    • 제31권1호
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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.