• Title/Summary/Keyword: anti-aggregation activity

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Characterization of Homocysteine ${\gamma}$-Lyase from Submerged and Solid Cultures of Aspergillus fumigatus ASH (JX006238)

  • El-Sayed, Ashraf S.;Khalaf, Salwa A.;Aziz, Hani A.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.499-510
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    • 2013
  • Among 25 isolates, Aspergillus fumigatus ASH (JX006238) was identified as a potent producer of homocysteine ${\gamma}$-lyase. The nutritional requirements to maximize the enzyme yield were optimized under submerged (SF) and solid-state fermentation (SSF) conditions, resulting in a 5.2- and 2.3-fold increase, respectively, after the last purification step. The enzyme exhibited a single homogenous band of 50 kDa on SDS-PAGE, along with an optimum pH of 7.8 and pH stability range of 6.5 to 7.8. It also showed a pI of 5.0, as detected by pH precipitation with no glycosyl residues. The highest enzyme activity was obtained at $37-40^{\circ}C$, with a $T_m$ value of $70.1^{\circ}C$. The enzyme showed clear catalytic and thermal stability below $40^{\circ}C$, with $T_{1/2}$ values of 18.1, 9.9, 5.9, 3.3, and 1.9 h at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. Additionally, the enzyme $K_r$ values were 0.002, 0.054, 0.097, 0.184, and 0.341 $S^{-1}$ at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. The enzyme displayed a strong affinity to homocysteine, followed by methionine and cysteine when compared with non-S amino acids, confirming its potency against homocysteinuria-related diseases, and as an anti-cardiovascular agent and a specific biosensor for homocysteinuria. The enzyme showed its maximum affinity for homocysteine ($K_m$ 2.46 mM, $K_{cat}\;1.39{\times}10^{-3}\;s^{-1}$), methionine ($K_m$ 4.1 mM, $K_{cat}\;0.97{\times}10^{-3}\;s^{-1}$), and cysteine ($K_m$ 4.9 m M, $K_{cat}\;0.77{\times}10^{-3}\;s^{-1}$). The enzyme was also strongly inhibited by hydroxylamine and DDT, confirming its pyridoxal 5'-phosphate (PLP) identity, yet not inhibited by EDTA. In vivo, using Swiss Albino mice, the enzyme showed no detectable negative effects on platelet aggregation, the RBC number, aspartate aminotransferase, alanine aminotransferase, or creatinine titer when compared with negative controls.

The Effect of Eisenia bicyclis Extracts on Antioxidant Activity and Serum Lipid Level in Ovariectomized Rats (대황 추출물의 in vitro 항산화 활성 및 난소를 절제한 흰쥐의 혈중 지질함량에 미치는 영향)

  • Park, Yong Soo;Kim, Mihyang
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1407-1414
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    • 2012
  • Hormone replacement therapy (HRT) is an effective regimen that has been found to prevent these diseases in postmenopausal women. However, HRT is accompanied by an increased risk of unfavorable outcomes. This study was conducted to evaluate the effects of Eisenia Bicyclis extract on lipids in ovariectomized rats. Fifty 7-week-old female Sprague-Dawley rats were randomly assigned to four groups: sham-operated rats (SHAM), ovariectomized rats (OVX-CON), and ovariectomized rats that were treated with Eisenia bicyclis extracts. The extract-treated diets were fed to the rats for 6 weeks after operation. Antioxidant effects were measured by DPPH free radical scavenging activity. Antioxidant activity of the ethanol extract increased in a dose-dependent manner and was about 55.9% in a concentration of 100 ${\mu}g/ml$. We measured the total cholesterol content, triglyceride content, HDL-cholesterol content, LDL-cholesterol content, atherosclerotic index, cardiac risk factor in serum, and anti-platelet aggregation and blood rheology. The total cholesterol and triglyceride concentration in serum increased for the OVX-control group, but supplementation with the E. bicyclis extract caused these factors to decrease. Notably, the serum LDL-cholesterol concentration in the OVX-EB200 group was significantly lower than the OVX-CON group. In addition, the blood passage times in rats that received the E. bicyclis extract were more rapid than the times in the untreated group (OVX-CON). Microscopic evaluation revealed that whole blood passed more smoothly through the microchannels in rats in the E. bicyclis extract supplement groups. Our results clarified the effects of E. bicyclis extract on serum lipid content in ovariectomized rats, and consequently we expect positive effects from providing E. bicyclis extract to postmenopausal women with cardiovascular disease.

The Comparative Understanding between Red Ginseng and White Ginsengs, Processed Ginsengs (Panax ginseng C. A. Meyer) (홍삼과 백삼의 비교 고찰)

  • Nam, Ki-Yeul
    • Journal of Ginseng Research
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    • v.29 no.1
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    • pp.1-18
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    • 2005
  • Ginseng Radix, the root of Panax ginseng C. A. Meyer has been used in Eastern Asia for 2000 years as a tonic and restorative, promoting health and longevity. Two varieties are commercially available: white ginseng(Ginseng Radix Alba) is produced by air-drying the root, while red ginseng(Ginseng Radix Rubra) is produced by steaming the root followed by drying. These two varieties of different processing have somewhat differences by heat processing between them. During the heat processing for preparing red ginseng, it has been found to exhibit inactivation of catabolic enzymes, thereby preventing deterioration of ginseng quality and the increased antioxidant-like substances which inhibit lipid peroxide formation, and also good gastro-intestinal absorption by gelatinization of starch. Moreover, studies of changes in ginsenosides composition due to different processing of ginseng roots have been undertaken. The results obtained showed that red ginseng differ from white ginseng due to the lack of acidic malonyl-ginsenosides. The heating procedure in red ginseng was proved to degrade the thermally unstable malonyl-ginsenoside into corresponding netural ginsenosides. Also the steaming process of red ginseng causes degradation or transformation of neutral ginsenosides. Ginsenosides $Rh_2,\;Rh_4,\;Rs_3,\;Rs_4\;and\;Rg_5$, found only in red ginseng, have been known to be hydrolyzed products derived from original saponin by heat processing, responsible for inhibitory effects on the growth of cancer cells through the induction of apoptosis. 20(S)-ginsenoside $Rg_3$ was also formed in red ginseng and was shown to exhibit vasorelaxation properties, antimetastatic activities, and anti-platelet aggregation activity. Recently, steamed red ginseng at high temperature was shown to provide enhance the yield of ginsenosides $Rg_3\;and\;Rg_5$ characteristic of red ginseng Additionally, one of non-saponin constituents, panaxytriol, was found to be structually transformed from polyacetylenic alcohol(panaxydol) showing cytotoxicity during the preparation of red ginseng and also maltol, antioxidant maillard product, from maltose and arginyl-fructosyl-glucose, amino acid derivative, from arginine and maltose. In regard to the in vitro and in vivo comparative biological activities, red ginseng was reported to show more potent activities on the antioxidant effect, anticarcinogenic effect and ameliorative effect on blood circulation than those of white ginseng. In oriental medicine, the ability of red ginseng to supplement the vacancy(허) was known to be relatively stronger than that of white ginseng, but very few are known on its comparative clinical studies. Further investigation on the preclinical and clinical experiments are needed to show the differences of indications and efficacies between red and white ginsengs on the basis of oriental medicines.