Bai F W, Ge X M, Anderson W A, Moo-Young M
Department of Bioscience and Bioengineering, Dalian University of Technology, Dalian 116023, China.
Biotechnol Bioeng. 2009 Jan 1;102(1):113-21. doi: 10.1002/bit.22043.
A bioreactor system composed of a stirred tank and three tubular bioreactors in series was established, and continuous ethanol fermentation was carried out using a general Saccharomyces cerevisiae strain and a very high gravity medium containing 280 g L(-1) glucose, supplemented with 5 g L(-1) yeast extract and 3 g L(-1) peptone. Sustainable oscillations of glucose, ethanol, and biomass were observed when the tank was operated at the dilution rate of 0.027 h(-1), which significantly affected ethanol fermentation performance of the system. After the tubular bioreactors were packed with 1/2'' Intalox ceramic saddles, the oscillations were attenuated and quasi-steady states were achieved. Residence time distributions were studied for the packed bioreactors by the step input response technique using xylose as a tracer, which was added into the medium at a concentration of 20 g L(-1), indicating that the backmixing alleviation assumed for the packed tubular bioreactors could not be established, and its contribution to the oscillation attenuation could not be verified. Furthermore, the role of the packing's yeast cell immobilization in the oscillation attenuation was investigated by packing the tubular bioreactors with packings with significant difference in yeast cell immobilization effects, and the experimental results revealed that only the Intalox ceramic saddles and wood chips with moderate yeast cell immobilization effects could attenuate the oscillations, and correspondingly, improved the ethanol fermentation performance of the system, while the porous polyurethane particles with good yeast cell immobilization effect could not. And the viability analysis for the immobilized yeast cells illustrated that the extremely lower yeast cell viability within the tubular bioreactors packed with the porous polyurethane particles could be the reason for their inefficiency, while the yeast cells loosely immobilized onto the surfaces of the Intalox ceramic saddles and wood chips could be renewed during the fermentation, guaranteeing their viability and making them more efficient in attenuating the oscillations. The packing Raschig rings without yeast cell immobilization effect did not affect the oscillatory behavior of the tubular bioreactors, further supporting the role of the yeast cell immobilization in the oscillation attenuation.
建立了一个由搅拌罐和三个串联的管式生物反应器组成的生物反应器系统,并使用普通酿酒酵母菌株和含有280 g L⁻¹葡萄糖、添加5 g L⁻¹酵母提取物和3 g L⁻¹蛋白胨的高浓度培养基进行连续乙醇发酵。当搅拌罐以0.027 h⁻¹的稀释率运行时,观察到葡萄糖、乙醇和生物量的持续振荡,这显著影响了系统的乙醇发酵性能。在管式生物反应器中填充1/2英寸的英塔洛克斯陶瓷鞍形填料后,振荡减弱并达到准稳态。通过使用木糖作为示踪剂的阶跃输入响应技术研究了填充生物反应器的停留时间分布,木糖以20 g L⁻¹的浓度添加到培养基中,结果表明,对于填充管式生物反应器所假定的返混减轻并未实现,其对振荡衰减的贡献也无法得到验证。此外,通过用酵母细胞固定化效果有显著差异的填料填充管式生物反应器,研究了填料的酵母细胞固定化在振荡衰减中的作用,实验结果表明,只有酵母细胞固定化效果适中的英塔洛克斯陶瓷鞍形填料和木屑能够减弱振荡,并相应地提高系统的乙醇发酵性能,而酵母细胞固定化效果良好的多孔聚氨酯颗粒则不能。对固定化酵母细胞的活力分析表明,填充有多孔聚氨酯颗粒的管式生物反应器内酵母细胞活力极低可能是其效率低下的原因,而松散固定在英塔洛克斯陶瓷鞍形填料和木屑表面的酵母细胞在发酵过程中可以更新,保证了它们的活力,使其在减弱振荡方面更有效。没有酵母细胞固定化效果的拉西环填料不影响管式生物反应器的振荡行为,进一步支持了酵母细胞固定化在振荡衰减中的作用。