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Production of eurycomanone from cell suspension culture of Eurycoma longifolia

机译:从长叶Eurycoma的细胞悬浮培养物中生产eurycomanone

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Eurycomanone is found in the Eurycoma longifolia Jack (Simaroubaceae) tree, exhibits significant antimalarial activity, improves spermatogenesis, suppresses expression of lung cancer cell tumour markers and regulates signalling pathways involved in proliferation, cell death and inflammation. Establishment of cell suspension culture of E. longifolia to determine the eurycomanone accumulation during cultures. Callus of E. longifolia was cultured in MS medium supplemented with 0.8% agar, 30/L sucrose, 1.25?mg/L NAA and 1?mg/L KIN for biomass production. Cell suspension culture was established by transferring friable calli to the same medium without agar. Eurycomanone content during cell culture was determined by HPLC with a C18 column, flow rate of 0.8?mL/min, run time of 17.5?min, detector wavelength of 254?nm. The stationary phase was silica gel and the mobile phase was acetonitric:H2O. Roots of 5?year-old trees were used as the control. The cells from 3?g of inoculum increased in biomass with a maximum value of 16?g fresh weight (0.7?g dry weight) at 14th day of culture. The cell growth then decreased from day 14 to day 20. Eurycomanone was produced during culture from the beginning to 20th day, its highest content (1.7?mg/g dry weight) also obtained at 14th day (the control is 2.1?mg/g dry weight). Cell suspension culture of E. longifolia is a suitable procedure to produce eurycomanone. The yield of eurycomanone biosynthesis in 14?days-old cells are relatively high, approximately 0.8 times the control.
机译:Eurycomanone是在长叶Eurycoma Jackif(Simaroubaceae)树中发现的,具有显着的抗疟活性,改善精子生成,抑制肺癌细胞肿瘤标志物的表达并调节涉及增殖,细胞死亡和炎症的信号通路。建立E. longifolia细胞悬浮培养以确定在培养过程中eurycomanone的积累。在添加0.8%琼脂,30 / L蔗糖,1.25?mg / L NAA和1?mg / L KIN的MS培养基中培养长叶大肠杆菌的愈伤组织,以生产生物质。通过将易碎的愈伤组织转移到没有琼脂的相同培养基中来建立细胞悬浮培养物。用C18色谱柱通过HPLC测定细胞培养过程中的杜仲酮含量,流速为0.8?mL / min,运行时间为17.5?min,检测器波长为254?nm。固定相为硅胶,流动相为乙腈:H2O。以5岁树的根为对照。在培养的第14天,来自3克接种物的细胞的生物量增加,最大值为16克鲜重(0.7克干重)。然后从第14天到第20天,细胞生长下降。从培养开始到第20天,在生产过程中产生了杜仲酮,在第14天也获得了其最高含量(1.7?mg / g干重)(对照组为2.1?mg / g净重)。 E. longifolia的细胞悬浮培养是生产eurycomanone的合适方法。在14天大的细胞中,乙草酮的生物合成产量相对较高,约为对照的0.8倍。

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