• Title/Summary/Keyword: protein oxidation

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Rapid Screening Method of Peroxidase by Colorimetric Assay and Screening of 2, 4-DCP Degradable Strains (발색법에 의한 Peroxidase의 신속한 스크리닝법과 2, 4-DCP 분해균주의 스크리닝)

  • Ryu, Kang;Lee, Eun-Kyu
    • KSBB Journal
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    • v.23 no.6
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    • pp.484-488
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    • 2008
  • Chlorinated phenols are widely used by the chemical industry as intermediate products in synthesis and previously were frequently applied to various industry fields. Peroxidases catalyze the peroxide-dependent oxidation of a range of inorganic and organic compounds. Peroxidase was shown to mineralize a variety of recalcitrant aromatic compounds and to oxidize a number of polycyclic aromatic and phenolic compounds. Among monomeric phenolic and nonphenolic compounds, peroxidase is known to oxidize its compounds. In this study, a colorimetric assay was developed to quantitatively evaluate the peroxidase activity for rapid screening. Color products of different intensity were developed proportionally to the peroxidase activity on agar plate and 96-well plate. This method correlates well with the RP-HPLC result. Using this screening method, 12 colonies of strain was screened which survived at high concentration of 2,4-DCP (1000 ppm) and with peroxidase activity for the $7^{th}$ round screening step on agar plate. These strains were utilized 2,4-DCP as a sole carbon source and produced peroxidase. After the screening test, four of the bacteria have significant better effect of COD removal on dye waste-water. COD removal of these was from 44% to 61%, respectively.

Peroxidase Activity during Leaf Infection of Mulberry (Morus alba L.) with Brown Leaf Spot Fungus Myrothecium roridum

  • Chattopadhyay Soumen;Krishnan Natraj;Maji Manas D.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.12 no.1
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    • pp.21-28
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    • 2006
  • Peroxidase activity was measured in brown leaf spot pathogen (Myrothecium roridum) inoculated potted mulberry (Morus alba) during pre-symptomatic to various symptom development stages and compared with corresponding healthy leaf tissues. The enzyme showed a pH optimum of 7.0 and the activity was linearly increased up to 15 min of incubation. The peroxidase had a broad substrate specificity and the rates of oxidation were in the rank of pyrogallol> guaiacol> ascorbate at pH 7.0. Catechol at 10 mM inhibited 89% of guaiacol-peroxidase and 76% pyrogallol-peroxidase activities, indicated higher non-specific peroxidation in pyrogallol dependent assay system in mulberry than guaiacol. The optimum requirement for the guaiacol dependent assay was 0.2 ml (${\approx}40-60{\mu}g$ equivalent of protein) of crude enzyme source. Excepting the 8th leaf from the apex, the peroxidase activity did not vary appreciably in different leaf positions. In pre-symptomatic phases, an initial (1 to 5 min) rise of peroxidase activity was noticed in inoculated leaves, and then maintained a plateau up to 300 min. In contrary, non-infected tissue showed a slightly increased trend of enzyme level up to 420 min. In infected tissue, a sharp transient increase (3.1 fold) of peroxidase activity appeared between 300 - 420 min post infections. Afterwards, significantly different but steady maintenance of enzyme levels were observed in two treatments. On the other hand, during symptom development, a sharp increase in peroxidase activity was noticed up to 4th grade of lesion appearance (25.1 % to 50% of leaf area infection), and then declined slightly. However, in non-infected but same age healthy leaves, such huge fluctuations of enzyme level did not apparent. A high positive correlation $(R^2=0.92)$ between peroxidase activity and leaf spot development grades was also marked. The result implies that pre-symptomatic burst (between 1 - 5 and 300 - 420 min) and subsequent increased trend of guaiacol peroxidase activity may require for the symptomatic manifestation of Myrothecium leaf spot in mulberry.

Self-Sufficient Catalytic System of Human Cytochrome P450 4A11 and NADPH-P450 Reductase

  • Han, Song-Hee;Eun, Chang-Yong;Han, Jung-Soo;Chun, Young-Jin;Kim, Dong-Hyun;Yun, Chul-Ho;Kim, Dong-Hak
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.156-161
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    • 2009
  • The human cytochrome P450 4A11 is the major monooxygenase to oxidize the fatty acids and arachidonic acid. The production of 20-hydroxyeicosatetraenoic acid by P450 4A11 has been implicated in the regulation of vascular tone and blood pressure. Oxidation reaction by P450 4A11 requires its reduction partners, NADPH-P450 reductase (NPR). We report the functional expression in Escherichia coli of bicistronic constructs consisting of P450 4A11 encoded by the first cistron and the electron donor protein, NPR by the second. Typical P450 expression levels of wild type and several N-terminal modified mutants was observed in culture media and prepared membrane fractions. The expression of functional NPR in the constructed P450 4A11: NPR bicistronic system was clearly verified by reduction of nitroblue tetrazolium. Membrane preparation containing P450 4A11 and NPR efficiently oxidized lauric acid mainly to $\omega$-hydroxylauric acid. Bicistronic coexpression of P450 4A11 and NPR in E. coli cells can be extended toward identification of novel drug metabolites or therapeutic agents involved in P450 4A11 dependent signal pathways.

DC23, a Triazolothione Resorcinol Analogue, Is Extensively Metabolized to Glucuronide Conjugates in Human Liver Microsomes

  • Shon, Jong Cheol;Joo, Jeongmin;Lee, Taeho;Kim, Nam Doo;Liu, Kwang-Hyeon
    • Mass Spectrometry Letters
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    • v.9 no.1
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    • pp.24-29
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    • 2018
  • DC23, a triazolothione resorcinol analogue, is known to inhibit heat shock protein 90 and pyruvate dehydrogenase kinase which are up-regulated in cancer and diabetes, respectively. This study was performed to elucidate the metabolism of DC23 in human liver microsomes (HLMs). HLMs incubated with DC23 in the presence of uridine 5'-diphosphoglucuronic acid (UDPGA) and/or ${\beta}$-nicotinamide adenine dinucleotide phosphate (NADPH) resulted in the formation of four metabolites, M1-M4. M1 was identified as DC23-N-Oxide, on the basis of LC-MS/MS analysis. DC23 was further metabolized to its glucuronide conjugates (M2, M3, and M4). In vitro metabolic stability studies conducted with DC23 in HLMs revealed significant glucuronide conjugation with a $t_{1/2}$ value of 1.3 min. The inhibitory potency of DC23 on five human cytochrome P450s was also investigated in HLMs. In these experiments, DC23 inhibited CYP2C9-mediated tolbutamide hydroxylase activity with an $IC_{50}$ value of $8.7{\mu}M$, which could have implications for drug interactions.

Improving effect of Artemisiae Capillaris Herba extract in reflux esophagitis rats (인진호 추출물의 급성역류성 식도염 유발 흰쥐에 대한 개선 효과)

  • Lee, Joo Young;Seo, Bu Il;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.31 no.6
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    • pp.37-44
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    • 2016
  • Objective : This study aimed to evaluate the protective effect of Artemisiae Capillaris Herba (AC) in reflux esophagitis (RE) rats. Methods : The AC was measured antioxidant activity through in vitro experiments, such as total polyphenol and flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Base on the results, we had conducted in vivo experiments. Rats were divided normal, control, AC treatment 50 mg/kg BW (AC50), and AC treatment 100 mg/kg BW (AC100) groups. AC were orally administered 2 h before the induction of RE. RE was induced by tie the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats. The rats were sacrificed 5 h after the surgery. We analyzed the expression of inflammatory related markers by western blot and observed the production of reactive oxygen species (ROS) and hematoxylin-eosin staining, Results : The $IC_{50}$ of AC for DPPH and ABTS were showed 12.60 and $33.32{\mu}g/m{\ell}$ respectively. In the RE rat, AC decreased inflammatory related markers, such as phosphorylated inhibitor of ${\kappa}B{\alpha}$, nuclear factor-kappa B, cyclooxygenase-2, inducible nitric oxide synthase, and tumor necrosis factor alpha. Also, AC reduced the increased reactive oxygen species in serum. The anti-inflammatory effect of AC appeared to be partially mediated through the inhibition of ROS. Also, AC markedly ameliorated esophageal mucosa damage via the inhibition of protein expression related to inflammation. Conclusions : Therefore, these results suggest that AC would be used as a therapeutic material in protection and/or treatment for reflux esophagitis.

Complete genome sequence of Betaproteobacteria strain GR16-43 isolated form a freshwater pond in South Korea (담수에서 분리한 Betaproteobacteria GR16-43의 유전체 염기서열 분석)

  • Choi, Ahyoung;Baek, Kiwoon;Chung, Eu Jin;Kim, Jee-Hwan;Choi, Gang-Guk
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.320-322
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    • 2017
  • A betaproteobacterium strain GR16-43 was isolated from a surface layer of the Geomnyong Pond in Republic of Korea by a dilution-to-extinction culturing method. We report the whole genome sequence of the strain GR16-43, which contains 4,806,848 bp with a G + C content 67.12%, and to include 4,424 protein-coding genes and 47 transfer RNA genes. The genome was determined to contain the genes encoding carbon monoxide dehydrogenase, nitrate reductase, nitrite reductase, nitric oxide reductase, and the sulfur oxidation (sox) gene cluster, highlighting the potential importance of the bacterial group represented by the strain in the cycling of inorganic elements. These results indicate that strain GR16-43 genome showed several traits indicating adaptation of the bacteria to living in freshwater environments.

Niacinamide Protects Skin Cells from Oxidative Stress Induced by Particulate Matter

  • Zhen, Ao Xuan;Piao, Mei Jing;Kang, Kyoung Ah;Fernando, Pincha Devage Sameera Madushan;Kang, Hee Kyoung;Koh, Young Sang;Yi, Joo Mi;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.27 no.6
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    • pp.562-569
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    • 2019
  • Niacinamide (NIA) is a water-soluble vitamin that is widely used in the treatment of skin diseases. Moreover, NIA displays antioxidant effects and helps repair damaged DNA. Recent studies showed that particulate matter 2.5 ($PM_{2.5}$) induced reactive oxygen species (ROS), causing disruption of DNA, lipids, and protein, mitochondrial depolarization, and apoptosis of skin keratinocytes. Here, we investigated the protective effects of NIA on $PM_{2.5}$-induced oxidative stress in human HaCaT keratinocytes. We found that NIA could inhibit the ROS generation induced by $PM_{2.5}$, as well block the $PM_{2.5}$-induced oxidation of molecules, such as lipids, proteins, and DNA. Furthermore, NIA alleviated $PM_{2.5}$-induced accumulation of cellular $Ca^{2+}$, which caused cell membrane depolarization and apoptosis, and reduced the number of apoptotic cells. Collectively, the findings show that NIA can protect keratinocytes from $PM_{2.5}$-induced oxidative stress and cell damage.

The Effect of Gyejibokryunghwan on Fracture Union in Tibia-fractured Rats (계지복령환(桂枝茯苓丸)이 흰쥐의 골절유합에 미치는 영향)

  • Yoon, Jeong-Won;Kim, Soon-Joong
    • Journal of Korean Medicine Rehabilitation
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    • v.29 no.4
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    • pp.1-14
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    • 2019
  • Objectives The object of this study was to assess the effect of Gyejibokryunghwan (GBH) on anti-oxidant and anti-inflammatory activities in RAW 264.7 cells and on factors associated with fracture union in tibia-fractured rats. Methods The 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity was measured to assess anti-oxidant activity. The production of nitric oxide (NO), interleukin-6 (IL-6), interleukin-$1{\beta}$ ($IL-1{\beta}$) and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to assess anti-inflammatory activity. The production of osteocalcin, calcitonin, carboxy-terminal telepeptides of type II collagen (CTXII), transforming growth factor-${\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2) in serum of tibia-fractured rats were measured to assess the effects of fracture union. X-rays were taken every two weeks from 0 to 4th week to assess fracture union effect. Results DPPH radical scavenging activity of GBH was increased according to concentration of GBH in RAW 264.7 cell. NO, prostaglandin $E_2$ ($PGE_2$), IL-6, $IL-1{\beta}$ and $TNF-{\alpha}$ were significantly decreased, indicating anti-inflammatory effect. Osteocalcin, calcitonin, $TGF-{\beta}$ were significantly increased in the experimental groups. CTXII was significantly decreased in the experimental groups. BMP-2 was not significantly changed in the experimental groups. The X-ray showed that the experimental group has better healing effects on tibia-fractured rats than control group. Conclusions From above result, GBH has an effect on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells. GBH showed significant results in factors related with fracture union and radiologic examination. In conclusion, GBH can help fracture union and it well be expected to be used actively in clinics.

Effects of natural nitrite source from Swiss chard on quality characteristics of cured pork loin

  • Kim, Tae-Kyung;Hwang, Ko-Eun;Song, Dong-Heon;Ham, Youn-Kyung;Kim, Young-Boong;Paik, Hyun-Dong;Choi, Yun-Sang
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1933-1941
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    • 2019
  • Objective: This study was conducted to evaluate quality characteristics of cured pork loin with natural nitrite source from Swiss chard. Methods: Pork loin was cured in brine and the ratio of water and fermented Swiss chard (FSC) solution in the brine was changed by 4:0 (control), 3:1 (T1), 1:1 (T2), 1:3 (T3), 0:4 (T4), and pickled samples with 0.012% sodium nitrite (PC, positive control) and nitrite free brine (NC, negative control) were considered as the control. Results: The pH values of cured pork loins with FSC were decreased with increasing addition level of FSC. Cooking loss was not significantly different among all treatments. T4 had the lowest value in moisture content and lightness value and the highest value in curing efficiency. The redness value of T4 was not significantly different from that of PC in raw. After cooking, however, it was higher than that of PC. The yellowness value of cured pork loin added with FSC was increased with increasing level of FSC. Volatile basic nitrogen content of cured pork loin added with FSC was higher than PC and NC. Thiobarbituric acid reactive substance value of cured pork loin added with FSC was decreased with increasing FSC level. Residual nitrite level and shear force were increased with increasing FSC level. In the sensory evaluation, sensory score for flavor, off-flavor, chewiness, juiciness, and overall acceptability were not significantly different among all treatments. However, sensory score for color was increased when the concentration of FSC added to pork loin was increased. Conclusion: The FSC solution had a positive effect on redness and lipid oxidation. As shown by the results in protein deterioration and sensory, Swiss chard can replace sodium nitrite as natural curing agent.

Large Area Deposition of Biomimetic Polydopamine-Graphene Oxide Hybrids using Langmuir-Schaefer Technique (랭뮤어-쉐퍼 기법 이용 생체모사 폴리도파민-산화그래핀 복합체 대면적 적층 기법 연구)

  • Kim, Tae-Ho;Song, Seok Hyun;Jo, Kyung-Il;Koo, Jaseung
    • Journal of Adhesion and Interface
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    • v.20 no.3
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    • pp.110-115
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    • 2019
  • Graphene oxide has been gathering interests as a way to exfoliate graphene. Since the oxidation group of graphene oxide can hydrogen bond with various functional groups, tremendous efforts have been actively conducted to apply various applications. However, graphene oxide alone cannot substantially possess the mechanical properties required for the practical application. Therefore, in this study, polydopamine, which is a bio-mimetic mussel protein-inspired material, was combined with graphene oxide to form a large-area composite membrane at the liquid-gas interface. In addition, the morphology of the polydopamine-graphene oxide composite thin film was also controlled to obtain a composite membrane having a nano-wrinkle structure. It can be expected to be used in the next generation seawater desalination membranes or carbon composites because it can form mechanically superior and sophisticated nanostructures.