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四种前体对胀果甘草细胞悬浮培养生产甘草黄酮的调控效果评价

The Effect of Precursor Feeding on Flavonoids Biosynthesis in Cell Suspension Cultures of Glycyrrhiza inflata Bat.

  • 摘要: 在胀果甘草细胞悬浮体系中加入苯丙氨酸、酪氨酸、肉桂酸和乙酸钠来研究前体对甘草细胞生产甘草黄酮的影响。结果显示,4种前体在合适的浓度下对细胞的生长没有明显的抑制作用,而且均能促进细胞内甘草黄酮的生物合成,但高浓度的肉桂酸对细胞的生长有一定的抑制作用。苯丙氨酸的最佳添加浓度为20 mg/L,酪氨酸、肉桂酸、乙酸钠的最佳添加浓度都是5 mg/L。此时,均可使培养体系的甘草黄酮产量高达100 mg/L以上,其中酪氨酸的添加使得产量高达对照的1.43倍。苯丙氨酸、肉桂酸和乙酸钠3种前体的添加时间均以第10天为宜,酪氨酸添加时间以第5天为最佳。而且,在添加苯丙氨酸和乙酸钠后的第3天收获细胞,此时细胞的生物量和甘草黄酮产量最大。此外,苯丙氨酸、乙酸钠的添加可以增加黄酮合成的关键酶之一——苯丙氨酸裂解酶的活性。酪氨酸对苯丙氨酸裂解酶影响不大,而肉桂酸的添加却导致其活性显著降低。

     

    Abstract: The effects of four precursors on flavonoids biosynthesis in cell suspension cultures of Glycyrrhiza inflata Bat.were studied.The results showed that all of these precursors with the appropriate concentrations not only affected the cell growth,but also promoted the flavonoids biosynthesis.Moreover,the flavonoids productions were all beyond 100 mg/L when the best addition concentrations of phenylalanine,tyrosine,cinnamic acid and NaOAc were 20,5,5 and 5 mg/L,respectively.The optimum time for tyrosine addition was the 5th day during the cell culture,while it was the 10th day for the other three kinds of precursors.Among the four precursors,tyrosine was the most effective in which the maximum flavonoids production reached 113.0 mg/L,1.43 times as that of the control.Additionally,the addition of phenylalanine and NaOAc enhanced the activity of phenylalanine ammonialyase,one of the key enzymes of the flavonoids biosynthesis,while the addition of cinnamic acid deduced it.

     

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