• 제목/요약/키워드: ABTS assay

검색결과 543건 처리시간 0.026초

Purification and Properties of Laccase of the White-rot Basidiomycete Coriolus hirsutus

  • Lee, Yeo-Jin;Shin, Kwang-Soo
    • Journal of Microbiology
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    • 제37권3호
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    • pp.148-153
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    • 1999
  • Laccase produced by Coriolus hirsutus was purified to electrophoretic homogeneity by acetone precipitation, Sephacryl S-2000 HR chromatography, DEAE Sepharose CL-6B chromatography, and Mono Q HR 5/5 chromatography. The purification of laccase was 46.6-fold with an overall yield of 23.7%. Laccase from this fungus was a monomeric glycoprotein with 16% carbohydrate content, and has an isoelectric point of 4.2, and molecular mass of 78 kDa, respectively. The N-terminal amino acid sequence of the enzyme showed significant homology to hoste of laccases from Coriolus versicolor, Pycnoporus cinnabarius, and an unidentified basidiomycete, PM1. The highest rate of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) oxidation by laccase was reached at 45$^{\circ}C$, and te pH optima of the enzyme varied depending on the substrate in the range of 2.0 to 4.5. The enzyme was stable at 60$^{\circ}C$ for 5 h and lost 80% activity at 80$^{\circ}C$ in 30 min. The enzyme oxidized a variety of usual laccase substrates including lignin-related phenol, and had the highest affinity toward ABTS. Under standard assay conditions, the apparent Km value of the enzyme toward ABTS was 8.1 ${\mu}$M. The enzyme was completely inhibited by L-cysteine and sodium azide, but not by potassium cyanide, SDS, ad thiourea.

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Anti-Oxidative and Anti-Neuroinflammatory Effect of Ethanol Extracts from Walnuts's (Juglans regia L.) Shell

  • Kang, Hyun
    • 대한의생명과학회지
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    • 제24권4호
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    • pp.365-371
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    • 2018
  • In this study, antioxidant and anti-neuroinflammatory of ethanol extracts from walnuts's (Juglans regia L.) shell were investigated in vitro. Radical-scavenging activities of the walnuts's shell ethanol extracts (WSE) were examined by using ABTS radicals and ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radicals assay. In the ABTS and DPPH radical scavenging activity, $RC_{50}$ of WSE were measured as 15.74 and $40.13{\mu}g/mL$, respectively. Also, to evaluate the anti-neuroinflammatory effects of WSE in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. The production of proinflammatory cytokines NO were examined by LPS in BV-2 cell. BV-2 cells activated with LPS were treated with various doses (10, 25, 50, $100{\mu}g/mL$) of WSE. Supernatants were analyzed for the production of NO using Griess reagent. WSE up to $10{\mu}g/mL$ still required to inhibit NO induced by LPS. These results showed that walnuts's (Juglans regia L.) shell can be used as an easily accessible source of natural anti-neuroinflammatory and natural antioxidants.

조리 과정 중 시금치의 항산화 활성 및 항균 활성의 변화 (Cooking Process for Spinach and Their Effects on Antioxidant and Antimicrobial Activities)

  • 박초희;김경희;태미화;김나영;육홍선
    • 한국식품영양학회지
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    • 제27권2호
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    • pp.147-155
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    • 2014
  • 시금치의 조리 과정(non-blanched, blanched, seasoned) 중 총 폴리페놀, 플라보노이드 함량 및 항산화 활성과 항균 활성 변화를 측정하였다. 80% 에탄올에 추출한 시금치의 수율은 데치기 전 1.64%, 데친 후 1.49%, 무친 후 6.01%로 무친 후 > 데치기 전 > 데친 후 순으로 높은 수율을 보였다. 총 폴리페놀 함량은 무친 후가 $124.31{\pm}1.37mg$ GAE/100 g FW로 가장 높았고, 데치기 전 $51.24{\pm}0.27mg$ GAE/100 g FW, 데친 후 $42.48{\pm}0.53mg$ GAE/100 g FW로 데친 후가 가장 낮은 총 폴리페놀 함량을 보였다. 총 플라보노이드 함량에서도 무친 후가 $15.60{\pm}0.20mg$ CHE/100 g FW로 가장 높은 함량을 나타냈다. 무친 후의 시금치는 4가지의 항산화 실험(DPPH assay, ABTS assay, FRAP assay, Reducing power)에서도 데치기 전이나 데친 후의 시금치보다 우수한 활성을 보였다. 항균 활성 측정 결과, 데치기 전 시금치는 S. enterica와 P. aeruginosa 두 개의 균에 대해서 5 mg/disc와 10 mg/disc의 두 농도에서 항균 활성을 갖고 있는 것으로 나타났으며, 데친 후 시금치는 항균 활성을 나타내지 않았고, 무친 후 시금치는 P. aeruginosa에 대해서만 10 mg/disc의 농도에서 8.15 mm의 clear zone을 형성하였다. 따라서 항균 활성 측면에서는 시금치를 생으로 섭취하는 것이 이로우나, 시금치를 생으로 과다 섭취할 경우, 체내에 결석이 생길 수 있으므로 끓는 물에 시금치를 데쳐 어느 정도의 수산을 제거하는 것이 바람직하다고 사료되며, 시금치를 무치는 과정에서 갖은 양념을 첨가함으로써 데치기 전보다 항산화 활성이 증가하므로, 가능한 무친 후의 시금치를 섭취하는 것이 건강적인 측면에서 이로울 것으로 사료된다.

채취시기별 보검선인장 줄기의 항산화, 항당뇨 및 항알츠하이머 활성평가 (Antioxidant and Anti-Diabetic, Anti-Alzheimer Activities of Stem from Opuntia ficus-indica var. saboten Cultivated in Jeju at Harvest Time)

  • 정윤숙;황병순;조수묵;황경아;황인국
    • 한국식품영양학회지
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    • 제30권6호
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    • pp.1332-1340
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    • 2017
  • In this study, we assessed antioxidant, anti-diabetic, and anti-Alzheimer activities of Opuntia ficus-indica var. saboten (OFI) at harvest time. OFIs were cultivated December 2015~November 2016 in Jeju island. The 70% ethanol extracts of OFI were used to investigate total polyphenol and flavonoid contents, antioxidant(DPPH and ABTS radical scavenging assay), anti-diabetic(yeast ${\alpha}$-glucosidase and rat ${\alpha}$-glucosidase inhibition assay), and anti-Alzheimer(Acetylcholinesterase and butyrylcholinesterase inhibition assay) activities. Total polyphenol and flavonoid contents of OFIs were $17.40{\sim}23.11{\mu}g$ garlic acid/mg Ex and 2.17~6.22 ug (+)-catechine/mg Ex, respectively. DPPH and ABTS radical scavenging activity of OFIs were 131.98~184.90 mg ascorbic acid(AA) eq/100 g and 63.60~101.83 mg AA eq/100 g, respectively. In the anti-diabetic and anti-Alzheimer activities, 70% ethanol extracts of OFI exhibited moderate inhibition activity, compared to control (acarbose and beberine). Total polyphenol and flavonoid contents, antioxidant, anti-diabetic, and anti-Alzheimer activities were no significant differences by season, respectively. Therefore, information on comparative biological evaluations of OFI may be a beneficial in exploring functional food and drug development.

접시꽃 캘러스 추출물의 천연화장품 원료로서의 효능 평가 (Evaluation of Alcea rosea L. Callus Extract as a Natural Cosmetic Ingredient)

  • 이기복;염아름;김동원;박창민;정민석;이기용;정철승
    • 대한화장품학회지
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    • 제44권3호
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    • pp.295-302
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    • 2018
  • 본 연구에서는 천연 화장품 소재 개발을 위하여 접시꽃 캘러스 생리활성 효능에 대해서 평가하였다. 접시꽃캘러스 추출물의 항산화 효능은 DPPH, ABTS, FRAP assay를 통해서 확인하였다. 그 결과 접시꽃캘러스 추출물은 농도 의존적으로 강력한 항산화 능력을 확인되었다. 더불어, 접시꽃 캘러스 추출물은 10 mg/mL 의 농도에서 세포 내 ROS를 효과적으로 감소시키는 것으로 확인되었다. 접시꽃 캘러스 추출물의 미백효능을 평가하기 위해 tyrosinase 저해 효과를 확인한 결과, 접시꽃 캘러스 추출물은 10 mg/mL 의 농도에서 tyrosinase 활성을 51% 감소시키는 것으로 확인되었다. 이상의 결과를 바탕으로, 접시꽃 캘러스 추출물은 항산화 및 미백효능을 지닌 화장품 기능성 소재로서 적용 가치가 높은 천연소재로 사료된다.

쥐오줌풀의 항산화 및 산화적 DNA 손상 억제 활성 (Antioxidative Activities and Inhibition Effects on Oxidative DNA Damage of Valeriana fauriei)

  • 박재호;장태원;이승현
    • 한국약용작물학회지
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    • 제24권6호
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    • pp.464-470
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    • 2016
  • Background: Valeriana fauriei (Valerianaceae) has been used to as a traditional medicine to treat a variety of symptoms, including headache, insomnia, hypertension, and menstrual irregularity. However, the present study investigates the species' antioxidant activity and its inhibition of oxidative DNA damage, which have yet to be studied. Methods and Results: The antioxidant activity was assessed using radical scavenging assays with 1,1-diphenyl-2-picryl hydrazyl (DPPH) and, 2, 2'-azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) and a reducing power assay. The total phenol content was also analyzed, and phenolic compounds were detected using HPLC/UV, whereas the inhibitory effect of Valeriana fauriei on oxidative DNA damage was measured using ${\phi}-174$ RF I plasmid DNA cleavage assay. The DPPH and ABTS radical scavenging activity were $75.17{\pm}3.55%$ and $95.83{\pm}0.63%$, repectively, and the reducing power was $93.14{\pm}1.74$ at $200{\mu}g/m{\ell}$. The total phenol content was $10.24{\pm}0.04mg/g$, whereas chlorogenic acid, catechin, caffeic acid and epicatechin were identified using HPLC/UV, and the ${\phi}-174$ RF I plasmid DNA cleavage assay indicated that V. fauriei provided protection against oxidative damage. Conclusions: The results of the present study suggest that V. fauriei has powerful antioxidant activity that can provide protective effects against the oxidative DNA damage caused by free radicals. The species, therefore, provides a valuable resource for the development of natural pharmaceutical to treat aging, cancer, and degenerative diseases.

사매 에탄올 추출물의 항산화 및 항염증 효과 (Antioxidative and Anti-inflammatory Effect of Ethanol Extract from Duchesnea chrysantha)

  • 이덕재;전인화;김현수;조일영;장선일
    • 동의생리병리학회지
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    • 제26권1호
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    • pp.59-66
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    • 2012
  • Oxidative stress has been implicated in cutaneous damage in various inflammatory skin diseases, including atopic dermatitis. The present study was undertaken to investigate the antioxidative and anti-inflammatory activities of the extract of Duchesnea chrysantha (DCE). DEC was prepared by extracting with 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Total flavonoid and polyphenol contents of DCE were included $24.73{\pm}0.45$ and $178.77{\pm}2.65$, respectively. DCE significantly increased electron donating ability (DPPH), nitrite scavenging (NO) and ABTS reducing activity in dose dependant. We investigated the anti-inflammatory effects of DCE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. DCE significantly suppressed NO and prdstaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with DCE in a dose dependent manner. These results suggest that DEC may has value as natural product with its high quality functional components, antioxidative and anti-inflammatory activities.

Antioxidant Activity of Glycyrrhiza cultivar Extracts

  • Kim, Minhee;Kang, Myunghoon;Kim, Wonnam
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 추계학술대회
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    • pp.88-88
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    • 2019
  • Antioxidants are involved in the defense mechanism against the attack of free radicals. This study was carried out to determine the antioxidant activity of new variety of Glycyrrhiza cultivar radix, Wongam and Sinwongam. Dissolved freeze dried Wongam and Sinwongam extracts were filtered by $0.2{\mu}m$ filter and serially diluted at the concentrations of $10{\mu}g/mL$, $50{\mu}g/mL$, $100{\mu}g/mL$, $500{\mu}g/mL$, and $1000{\mu}g/mL$. The antioxidant potential was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, ABTS (2,2-azino-bis (3-rthylbenzthiazoline-6-sulfonic acid) diammonium salt) radical cation decolorization assay, nitrite radical scavenging assay, and ferric reducing antioxidant power (FRAP) assay. DPPH radical scavenging activities (i.e. the highest value $50.9{\pm}0.8%$ by Wongam and $82.6{\pm}1.1%$ by Sinwongam), ABTS radical scavenging activities (i.e. the highest value $88.1{\pm}1.8%$ by Wongam and $98.6{\pm}0.1%$ by Sinwongam), and nitrite radical scavenging activities (i.e. the highest value $87.3{\pm}1.6%$ by Wongam and $89.8{\pm}0.8%$ by Sinwongam) increased in a dose-dependent manner. In addition, ferric reducing power activities also increased in a dose-dependent manner. The FRAP value of Wongam and Sinwongam extracts were $0.72{\pm}0.03$ and $0.99{\pm}0.06$ compared to ascorbic acid, as a positive control, was $1.32{\pm}0.02$. These results suggested that Wongam and Sinwongam have beneficial effects as a potent antioxidant.

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Reticulone, a Novel Free Radical Scavenger Produced by Aspergillus sp.

  • Ryoo, In-Ja;Xu, Guang-Hua;Kim, Young-Hee;Choo, Soo-Jin;Ahn, Jong-Seog;Bae, Ki-Hwan;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1573-1575
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    • 2009
  • Bioassay-guided fractionation of the culture broth of Aspergillus sp. FN070449 (KCTC 26428) using a DPPH (2,2-diphenyl-1-picrylhydrazyl) assay led to the isolation of two compounds: reticulone (1) and reticulol (2). Their chemical structures were elucidated on the basis of UV, IR, NMR, and MS spectroscopic analyses. Compound 1 exhibited more potent free radical scavenging activity on $ABTS^{\cdot+}$ (2,2'-azino-bis [3-ethylbenzthiazoline-6-sulphonic acid]) and DPPH radicals than did butylated hydroxyanisole (BHA) and caffeic acid.

Antioxidant Activity-of Curcuma Longa L., Novel Foodstuff

  • Choi, Hae-Yeon
    • Molecular & Cellular Toxicology
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    • 제5권3호
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    • pp.237-242
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    • 2009
  • The potential antioxidant activities of different fractions from methanolic extract of Curcuma longa L. were assayed in vitro. All of the fractions exception of n-hexane and $H_2O$ showed a strong antioxidant activity, especially the ethylacetate (EtOAc) fraction, which showed the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity ($IC_{50}=9.86\;{\mu}g/mL$). The results of 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay showed concentration dependency, the EtOAc fraction demonstrating a better result than the other fractions at the same concentration in this studies. Additionally, when the total phenolic contents of fractions were measured, the EtOAc fraction contained the highest level. Meanwhile, correlation analysis indicated a high correlation between the antiradical activity and the total phenolic contents, suggesting that fractions obtained from the methanolic extract of Curcuma longa L. have wide potential for use as sources of antioxidant material.