National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
Biotechnol Prog. 2011 Nov-Dec;27(6):1672-9. doi: 10.1002/btpr.681. Epub 2011 Aug 17.
Recently, cichoric acid production from hairy roots of Echinacea purpurea was significantly improved by ultrasound stimulation in an airlift bioreactor. In this article, the possible mechanism on ultrasound-intensified hairy root culture of E. purpurea in the bioreactor was elucidated with the help of computational fluid dynamics (CFD) simulation, membrane permeability detection, dissolved oxygen concentration detection, confocal laser-scanning microscopy (LSM) observation, and phenylalanine ammonium lyase (PAL) activity analysis. The CFD model developed in Part I was used to simulate the hydrodynamics and oxygen mass transfer in hairy root bioreactor culture stimulated by ultrasound. A dynamic mesh model combined with a changing Schmidt number method was used for the simulation of the ultrasound field. Simulation results and experimental data illustrated that ultrasound intensified oxygen mass transfer in the hairy root clump, which subsequently stimulated root growth and cichoric acid biosynthesis. Ultrasound increased the hairy root membrane permeability, and a high root membrane permeability of 0.359 h(-1) was observed at the bottom region in the bioreactor. LSM observation showed that the change in the membrane permeability recovered to normal in the further culture after ultrasound stimulation. PAL activity in the hairy roots was stimulated by ultrasound increase and was correlated well to cichoric acid accumulation in the hairy roots of E. purpurea.
最近,通过气升式生物反应器中的超声刺激,显著提高了西洋蒲公英发根中菊苣酸的产量。在本文中,借助计算流体动力学(CFD)模拟、膜通透性检测、溶解氧浓度检测、共聚焦激光扫描显微镜(LSM)观察和苯丙氨酸解氨酶(PAL)活性分析,阐明了超声增强生物反应器中 E. purpurea 发状根培养的可能机制。在第一部分中开发的 CFD 模型用于模拟超声刺激的发状根生物反应器培养中的流体动力学和氧传质。使用动态网格模型和变化的 Schmidt 数方法来模拟超声场。模拟结果和实验数据表明,超声增强了发状根团块中的氧传质,随后刺激了根的生长和菊苣酸的生物合成。超声增加了发根的膜通透性,在生物反应器的底部区域观察到高达 0.359 h(-1)的高根膜通透性。LSM 观察表明,超声刺激后,膜通透性的变化在进一步培养中恢复正常。超声刺激增加了发状根中的 PAL 活性,与 E. purpurea 发状根中菊苣酸的积累密切相关。