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首页> 外文期刊>Biotechnology Progress >Heterologous Expression of Vitreoscilla Hemoglobin (VHb) and Cultivation Conditions Affect the Alkaloid Profile of Hyoscyamus muticus Hairy Roots
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Heterologous Expression of Vitreoscilla Hemoglobin (VHb) and Cultivation Conditions Affect the Alkaloid Profile of Hyoscyamus muticus Hairy Roots

机译：玻璃藻血红蛋白（VHb）的异源表达和培养条件影响鼠李丝毛根的生物碱谱

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Fast-growing hairy root cultures of Hyoscyamus muticus induced by Agrobacterium rhizogenes offer a potential production system for tropane alkaloids.Oxygen deficiency has been shown to limit growth and biomass accumulation of hairy roots,whereas little experimental data is available on the effect of oxygen on alkaloid production.We have investigated the effect of Vitreoscilla hemoglobin (VHb) expression and cultivation conditions on the complete alkaloid profile of H.muticus hairy roots in shake flasks and in a laboratory scale bioreactor.We optimized the growth medium composition and studied the effects of sucrose,ammonium,nitrate,and phosphate on growth and alkaloid production.Maximum biomass accumulation was achieved with the highest and maximum hyoscyamine content with the lowest sucrose concentration.The optimum nitrate concentration for growth was higher for the VHb line than the control.Neither VHb expression nor aeration improved the hyoscyamine content significantly,thus suggesting that hyoscyamine biosynthesis is not limited by oxygen availability.Interestingly,the effect of VHb expression on the alkaloid profile was slightly different from that of aeration.VHb expression did not affect the concentrations of cuscohygrine,which was increased by aeration.Therefore,the effect of VHb is probably not related only to its ability to increase the intracellular effective oxygen concentration.
机译：发根农杆菌诱导的鼠尾草速生毛状根培养物为托烷生物碱提供了潜在的生产系统。氧缺乏已显示限制了毛状根的生长和生物量积累，而关于氧对生物碱的影响的实验数据很少。我们研究了玻璃藻血红蛋白（VHb）的表达和培养条件对摇瓶和实验室规模的生物反应器中H.muticus毛状根完整生物碱谱的影响。我们优化了生长培养基的组成并研究了蔗糖的影响，铵，硝酸盐和磷酸盐对生长和生物碱的生产。最大和最大的硫胺素含量，最低的蔗糖浓度实现了最大的生物量积累。VHb品系的最佳生长硝酸盐浓度高于对照。充气也不能显着提高硫胺素的含量，因此建议有趣的是，VHb表达对生物碱谱的影响与通气略有不同。VHb表达并不影响cuscohygrine的浓度，而是通过通气增加其浓度。因此，效果VHb的毒性可能不仅与其增加细胞内有效氧浓度的能力有关。

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