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[WAVE生物反应器中枇杷细胞悬浮培养生产熊果酸的研究]

[Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor].

作者信息

Li Hui-hua, Yao De-heng, Xu Jian, Wang Wei, Chang Qiang, Su Ming-hua

出版信息

Zhongguo Zhong Yao Za Zhi. 2015 May;40(9):1693-8.

Abstract

Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.

摘要

通过使用WAVE生物反应器对枇杷悬浮细胞进行放大培养,研究了细胞生长和熊果酸(UA)的积累。以细胞干重(DW)和UA含量为评价指标,在摇瓶和反应器中进行了对比试验。比较了1 L和5 L工作体积生物反应器中的培养基、DW和UA含量。在1 L工作体积的生物反应器中,开展了接种量、转速和旋转角度等主要因素的正交试验以寻找最佳组合。生物反应器中细胞生长的DW和UA含量分别比摇瓶中的高105.5%和27.65%。在生物反应器中,1 L和5 L工作体积的液体培养中元素的动态变化、细胞生长的干重和UA含量相似。接种量80 g、转速26 r·min(-1)、角度6°为最佳组合,在1 L工作体积的生物反应器中培养100 h后,细胞生物量达到19.01 g·L(-1),UA含量达到27.750 mg·g(-1)。WAVE生物反应器比摇瓶更适合枇杷细胞悬浮培养,且培养参数可从1 L放大到5 L。

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