• Title/Summary/Keyword: Oxidase

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Determination of Biogenic Amines using an Amperometric Biosensor with a Carbon Nanotube Electrode and Enzyme Reactor (Carbon Nanotube 전극과 효소반응기로 구성된 Amperometric Biosensor를 이용한 Biogenic Amines 검출)

  • Kim, Jong-Won;Jeon, Yeon-Hee;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.5
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    • pp.735-742
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    • 2010
  • Biogenic amines are synthesized by microbial decarboxylation for the putrefaction or fermentation of foods containing protein. Although biogenic amines such as histamine, tyramine, and putrescine are required for many physiological functions in humans and animals, consumption of high amounts of biogenic amines can cause toxicological effects, including serious gastrointestinal, cutaneous, hemodynamic, and neurological symptoms. In this study, a novel amperometric biosensor wasdeveloped to detect biogenic amines. The biosensor consisted of a working electrode, a reference electrode, a counter electrode, an enzyme reactor with immobilized diamine oxidase, an injector, a peristaltic pump and a potentiostat. A working electrode was fabricated with a glassy carbon electrode (GCE) by coating functionalized multi-walled carbon nanotubes (MWCNT-$NH_2$) and by electrodepositing Prussian blue (PB) to enhance electrical conductivity. A sensor system with PB/MWCNT-$NH_2$/GCE showed linearity in the range of $0.5 {\mu}M{\sim}100 {\mu}M$ hydrogen peroxide with a detection limit of $0.5 {\mu}M$. The responses for tyramine, 2-phenylethylamine, and tryptamine were 95%, 75%, and 70% compared to that of histamine, respectively. These results imply that the biosensor system can be applied to the quantitative measurement of biogenic amines.

Ginsenoside Rg1 suppresses early stage of adipocyte development via activation of C/EBP homologous protein-10 in 3T3-L1 and attenuates fat accumulation in high fat diet-induced obese zebrafish

  • Koh, Eun-Jeong;Kim, Kui-Jin;Choi, Jia;Jeon, Hui Jeon;Seo, Min-Jung;Lee, Boo-Yong
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.23-30
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    • 2017
  • Background: Ginsenoside Rg1 is a class of steroid glycoside and triterpene saponin in Panax ginseng. Many studies suggest that Rg1 suppresses adipocyte differentiation in 3T3-L1. However, the detail molecular mechanism of Rg1 on adipogenesis in 3T3-L1 is still not fully understood. Methods: 3T3-L1 preadipocyte was used to evaluate the effect of Rg1 on adipocyte development in the differentiation in a stage-dependent manner in vitro. Oil Red O staining and Nile red staining were conducted to measure intracellular lipid accumulation and superoxide production, respectively. We analyzed the protein expression using Western blot in vitro. The zebrafish model was used to investigate whether Rg1 suppresses the early stage of fat accumulation in vivo. Results: Rg1 decreased lipid accumulation in early-stage differentiation of 3T3-L1 compared with intermediate and later stages of adipocyte differentiation. Rg1 dramatically increased CAAT/enhancer binding protein (C/EBP) homologous protein-10 (CHOP10) and subsequently reduced the $C/EBP{\beta}$ transcriptional activity that prohibited the initiation of adipogenic marker expression as well as triglyceride synthase. Rg1 decreased the expression of extracellular signal-regulated kinase 1/2 and glycogen synthase kinase $3{\beta}$, which are also essential for stimulating the expression of $CEBP{\beta}$. Rg1 also reduced reactive oxygen species production because of the downregulated protein level of nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase 4 (NOX4). While Rg1 increased the endogenous antioxidant enzymes, it also dramatically decreased the accumulation of lipid and triglyceride in high fat diet-induced obese zebrafish. Conclusion: We demonstrated that Rg1 suppresses early-stage differentiation via the activation of CHOP10 and attenuates fat accumulation in vivo. These results indicate that Rg1 might have the potential to reduce body fat accumulation in the early stage of obesity.

Effect of hot water and ethanol extracts from Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production in adipogenesis of 3T3-L1 cells (3T3-L1 세포분화 중 지방축적 및 ROS 생성에 대한 연화 열수 및 에탄올 추출물의 효과)

  • Oh, Ji-Won;Lee, Jin-Ha;Lee, Ok-Hwan;Kim, Kye-Hoon;Kim, Hey-Ran;Lee, Hyo-Ku
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.744-750
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    • 2015
  • The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

Isolation and Identification of a Marine Bacterium, Pseudomonas sp. BK1 Producing Extracellular Enzymes Capable of Decomposing Multiple Complex Polysaccharides (복합 다당류 분해 효소들을 생산하는 해양미생물 Pseudomonas sp. BK1의 분리 및 특성)

  • Kim, Beom-Kyu;Jeon, Beong-Sam;Cha, Jae-Young;Park, Jeong-Won;Kim, Sam-Woong;Kim, Ji-Yoon;Park, Yong-Lark;Cho, Young-Su;Song, Jae-Young
    • Journal of Life Science
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    • v.13 no.6
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    • pp.871-878
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    • 2003
  • A marine bacterium (strain BKl) that produces extracellular enzymes capable of decomposing complex polysac-charides, such as agar, chitin, carboxymethylcellulose, xylan and mannan, was isolated from the marine red alga Porphyra dentata. Strain BKl was gram-negative, aerobic, catalase- and oxidase-positive, polarly flagellated bacilli that produce gelatinase and urease, but not decarboxylases. The G+C content of the DNA was 51.6 mol%. The major isoprenoid quinone component was identified as an ubiquinone-8, and the major cellular fatty acids were C16:0, C16:1 w6c and C18:1 w7c. Comparative 16S rRNA sequence analysis placed strain BK1 with members of the genus Pseudomonas. On the basis of phenotypic and genotypic data, the strain BK1 was shown to be a member of the subgroup of Pseudomonas, and named as Pseudomonas sp. BK1.

Effects of Hanbag Mushroom(Grifola frondosa) on Oxidative Stress in Diabetic Rats (당뇨유발 흰쥐에 있어서 산화적 스트레스에 대한 함박잎새버섯의 효과)

  • Lee, Soon-Yi;Lee, Chang-Yun;Park, Yeong-Chul;Kim, Jong-Bong
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1571-1575
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    • 2007
  • This research was carried out to investigate the effects of Hambag mushroom on the oxidative stress in diabetic rats, Sprague-Dawley. The diabetic rats induced by streptozotocin were fed with hambag mushroom-powder(G. frondosa) for 6 weeks. For the level of oxidative stress in liver and pancreas tissues, it was studied by measuring LPO (lipid oxide) level as an indicator of lipid peroxidation, XOD(xanthine oxidase) as one of important sources for free radicals and the levels of GSH and GST as anti-oxidant systems. Also, as an indicator of liver damaged by oxidative stress, the activities of serum ALT and AST were measured. It was observed that the levels of ALT, AST, LPO and XOD were higher by about two times in both tissues from diabetic rats than in those from control rats. This indicates that the oxidative stress induced by diabetes caused the tissues damages. However, these levels were decreased in the tissues from rats with hambag mushroom-powder. Futhermore, the activity of GST were higher in both tissues from diabetic rats fed with hambag mushroom-powder than in those from diabetic rats. Thus, it is considered that the hambag mushroom-powder decreases the level of oxidative stress by increasing activity of anti-oxidant system such as GSH and GST. It is suggested that the hambag mushroom-powder can be useful for preventing the tissues damaged by diabetes-induced oxidative stress.

Exercise and Reactive Oxygen Species (운동과 활성산소)

  • Kim, Hye Jin;Lee, Won Jun
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1078-1085
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    • 2017
  • Free radicals have long been considered damaging to various tissues. An excessive amount of reactive oxygen species (ROS) is known to have detrimental effects on the body and to be linked to numerous pathological conditions, such as cardiovascular disease, cancer, diabetes, and skeletal muscle atrophy. On the other hand, recent findings suggest that ROS is important for maintenance and development of cellular activity. Cells respond to increased oxidative stress by adaptive changes in the expression of a variety of proteins involved in the maintenance of cellular integrity. ROS is also essential for skeletal muscle function and metabolism. It is well known that physical exercise has many health benefits. Paradoxically, physical exercise also stimulates the production of ROS, which result in oxidative stress. Based on evidence amassed in the past decade, exercise itself may be considered an antioxidant because training increases the expression of antioxidant enzymes. In this review, we discuss the processes underlying the generation of ROS and its role in exercise-induced adaptation based on recent evidence. Furthermore, we discuss the possible role of NADPH oxidase in exercise-induced activation of insulin signaling and its effect on longevity.

Black Sesame Ethanolic Extract Promotes Melanin Synthesis (Melanin 합성을 촉진하는 흑임자 에탄올추출물의 효능)

  • Jeon, Sojeong;Kim, Moon-Moo
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1452-1461
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    • 2017
  • Melanin production by melanocytes in human hair follicles decreases with time and leads to the graying process, which is a phenotype of human aging and an index of aging. The reduction in melanin production is the result of decreased tyrosinase activity in hair follicles and an accumulation of active oxygen species, such as hydrogen peroxide. This study investigated antioxidant effects and melanin-promoting effects in B16F1 cells treated with black sesame ethanolic nonpolar-soluble extract (SBEEO) and black sesame ethanolic polar-soluble extract (SBEEP). In antioxidation experiments, both SBEEP and SBEEO did not eliminate 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical, but SBEEO at $64{\mu}g/ml$ showed low reducing power. SBEEP exerted cytotoxic effects at concentrations greater than $8{\mu}g/ml$, whereas SBEEO showed cytotoxic effects at concentrations greater than $4{\mu}g/ml$. SBEEP and SBEEO induced melanin synthesis, tyrosinase activity, and DOPA oxidase activity in vitro. In live cells, melanin synthesis was greater in the SBEEP treatment group as compared with that in the SBEEO treatment group. SBEEP stimulated melanin synthesis by modulating the expression of tyrosinase-related protein-2 (TRP-2), which is an important enzyme in melanin synthesis. These results imply that SBEEP obtained from black sesame ethanolic extract may have the potential to improve melanin synthesis.

Effects of the Spatholobi calulis extract on Antioxidation and Inhibition of Matrix Metalloproteinase in Human Skin Fibroblasts (계혈등 추출물의 항산화와 사람 피부 섬유아세포에서의 Matrix Metalloproteinase-1 발현저해 효과)

  • Sim Gwan Sub;Kim Jin Hwa;Lee Dong Hwan;Park Sung Min;Pyo Hyeong Bae;Zhang Yong He;Lee Bum Chun
    • KSBB Journal
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    • v.20 no.1 s.90
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    • pp.40-45
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    • 2005
  • The production of matrix metalloproteinases (MMPs) by the UV irradiated skin fibroblast and the degradation of exracellular matrix (ECM) by these enzymes is known as one of the main reasons of photoaging. In this study, to investigate the relationship between aging and Spatholobi caulis extract, we examined the effects of antioxidant, in vitro MMP inhibition and expression of UVA-induced MMP-1 in human dermal fibroblasts. Spathoiobi caulis extract was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $45.81{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $3.11{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Spatholobi caulis extract inhibited the activities of MMP-1 in a does-dependent manner and the IC50 value calculated from semi-log plots was $31.96{\mu}g/ml$. Also, UVA induced MMP expression was reduced $74.66\%$ by treatment with Spatholobi caulis extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore Spatholobi caulis extract was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that Spatholobi caulis extract may act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

Bacillus ginsengihumi sp. nov., a Novel Species Isolated from Soil of a Ginseng Field in Pocheon Province, South Korea

  • Ten Leonid N.;Im Wan-Taek;Baek Sang-Hoon;Lee, Jung-Sook;Oh, Hee-Mock;Lee, Sung-Taik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1554-1560
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    • 2006
  • A Gram-positive, aerobic or facultative anaerobic, non motile, endospore-forming bacterial strain, designated Gsoil $114^T$, was isolated from a soil sample of a ginseng field in Pocheon Province (South Korea), and was characterized taxonomically by using a polyphasic approach. It grew well on nutrient agar medium and utilized a limited number of organic substrates as sole carbon sources, including D-xylose and some other carbohydrates, but did not utilize L-amino acids and organic acids. The isolate was positive for oxidase test but negative for catalase, and negative for degradation of macromolecules such as starch, cellulose, xylan, casein, chitin, and DNA. The G+C content of the genomic DNA was 41.8 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were $anteiso-C_{15:0}$ (32.1%), $iso-C_{15:0}$ (30.5%), and $anteiso-C_{17:0}$ (30.2%). Comparative 16S rRNA gene sequence analysis showed that strain Gsoil $114^T$ fell within the radiation of the cluster comprising Bacillus species and joined Bacillus shackletonii LMG $18435^T$ with a bootstrap value of 95%. The highest 16S rRNA gene sequence similarities were found with Bacillus shackletonii LMG $18435^T$ (97.6%), Bacillus acidicola DSM $14745^T$ (96.9%), Bacillus sporothermodurans DSM $10599^T$ (96.5%), and Bacillus oleronius DSM $9356^T$ (96.5%). The phylogenetic distance from any other validly described species within the genus Bacillus was less than 96%. DNA-DNA hybridization experiments showed that the DNA-similarities between strain Gsoil $114^T$ and closest phylogenetic neighbors were less than 39%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $114^T$ (=KCTC $13944^T$=DSMZ $18134^T$) was classified in the genus Bacillus as the type strain of a novel species, for which the name Bacillus ginsengihumi sp. nov. is proposed.

Extracellular Polysaccharide Produced by a New Methylotrophic Isolate (새로운 메탄올 자화세균이 생산하는 세포외 다당류)

  • Lee, Ho J.;Kim, Si W.;Kim, Young M.
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.212-218
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    • 1998
  • An obligately methylotrophic bacterium which produces extracellular polysaccharide (EPS) was isolated through methanol-enrichment culture technique. The isolate was aerobic, nonmotile, and gram negative rod and exibited catalase, but no oxidase, activity. Plasmid, carotenoid, and poly-${\beta}$-hydroxybutyric acid were not found. The guanine plus cytosine content of DNA was 52-56%. The isolate was found to grow only on methanol and monomethylamine. Growth was optimal ($t_d=2.4h$) at $35^{\circ}C$ and pH 6.5 in a mineral medium containing 0.5% (v/v) methanol, 25 mM phosphate, and 0.212% ammonium sulfate. Methanol was assimilated through the ribulose monophosphate pathway. Maximun amount of EPS was produced in cells growing at the mid-stationary growth phase at $30^{\circ}C$ in a mineral medium (PH 6.5) containing 1.0% (v/v) methanol in the CIN ratio of 54.7. Thin-layer chromatographic and high performance liquid chromatographic analysis revealed that the EPS was composed of glucose and galactose. EPS which was not treated with ethanol (Pbe) exhibited stable viscosity under various concentrations of salts and temperatures hut showed high viscosity at low pH. EPS precipitated with ethanol (Pae) was found to be more stable in viscosity than the Pbe at various salt concentrations, temperatures, and pH. The Pae also exhibited higher viscosity than the Pbe and xanthan gum. Scanning electron microscopy revealed that the lyophilized Pbe and Pae have a multi-layered structure and a structure of thick fibers, respectively.

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