波浪式WAVE生物反应器悬浮培养枇杷细胞生产熊果酸的研究

采用波浪式WAVE生物反应器放大培养枇杷悬浮细胞,研究细胞生长及熊果酸(UA)的累积情况.摇瓶和反应器中,以细胞的干重和UA的含量为指标进行对比试验;反应器中1 L和5 L工作体积下,以培养液中元素代谢、细胞的干重、UA的含量为指标进行对比试验;生物反应器中,1 L的工作体积下,以接种量、转速、转动角度为因素开展正交试验筛选最佳的组合.反应器中细胞的干重增长及UA含量比摇瓶中高出了105.5%,27.65%;生物反应器中不同的工作体积下,培养液中元素的代谢、细胞的干重及UA的累积情况相似;生物反应器中,接种量为80 g,转速为26 r·min^-1,角度为6°是最佳组合,接种后100 h收获,细胞的干重为19.01 g·L^-1,UA质量分数为27.750 mg·g^-1.结果表明WAVE生物反应器比摇瓶更适合枇杷细胞悬浮培养,并且可实现1 L到5 L的培养放大.

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

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.