van Kleeff B H, Kuenen J G, Heijnen J J
Department of Microbiology and Enzymology, Delft University of Technology, The Netherlands.
Biotechnol Prog. 1996 Jul-Aug;12(4):510-8. doi: 10.1021/bp960033b.
Heat measurements have been successfully as an analytical tool for the study of the dynamics of energy metabolism of Saccharomyces cerevisiae and Candida utilis grown in continuous culture under fluctuating substrate supply. A low average dilution rate (D = 0.05 h-1) was maintained either by adding the medium as continuously (dropwise) as possible or (blockwise) by adding the medium at high speed during a short period (D = 0.5 h-1 for 40 s) and not at all during the following period (D = 0.00 h-1 for 360 s). The resulting biological activity was monitored on-line with conventional (O2 and CO2) off-gas analyses, DOT measurements, and heat flux measurements. In C. utilis cultures, the biomass-specific maximum oxygen consumption rate (qO2,max), the biomass yield (Ys,x), and the dynamic responses to a glucose pulse and to a change in feeding regime were not significantly affected by different preceding feeding regimes. In contrast, S. cerevisiae grown in continuous culture with blockwise feed showed a 50% increase in qO2,max and a 25% drop in Ys,x compared to the culture grown with dropwise feed. The dynamic response to a glucose pulse (0.6 g L-1) was slower for the continuous (dropwise) than for the blockwise grown S. cerevisiae. With a second testing method for the dynamic response of the yeasts, the feeding regime was changed. The blockwise fed S. cerevisiae proved to be better "trained" to cope with sudden changes in glucose supply and, therefore, was more "shockproof" toward a change in feeding regime. This clearly points to major differences in the intracellular metabolic flux control between the yeasts. These findings are of relevance for industrial baker's yeast production, where reactor mixing times of one to several minutes are not uncommon. The observed, heat production, together with the dissolved oxygen concentration, appeared to give the fastest response to actual changes in the culture. It is suggested that heat measurements can be a very useful tool to monitor and control the growth of S. cerevisiae in laboratory and industrial fermenter operations.
热量测量已成功用作分析工具,用于研究在波动底物供应条件下连续培养的酿酒酵母和产朊假丝酵母的能量代谢动力学。通过尽可能连续(逐滴)添加培养基,或(逐块)在短时间内高速添加培养基(40秒内D = 0.5 h-1),随后一段时间完全不添加(360秒内D = 0.00 h-1),维持较低的平均稀释率(D = 0.05 h-1)。通过常规的(O2和CO2)尾气分析、溶解氧测量和热通量测量对产生的生物活性进行在线监测。在产朊假丝酵母培养物中,不同的先前进料方式对生物质特异性最大耗氧率(qO2,max)、生物质产量(Ys,x)以及对葡萄糖脉冲和进料方式变化的动态响应没有显著影响。相比之下,与逐滴进料培养的酿酒酵母相比,逐块进料连续培养的酿酒酵母qO2,max增加了50%,Ys,x下降了25%。连续(逐滴)培养的酿酒酵母对葡萄糖脉冲(0.6 g L-1)的动态响应比对逐块培养的酿酒酵母慢。采用第二种酵母动态响应测试方法时,改变了进料方式。结果表明,逐块进料的酿酒酵母在应对葡萄糖供应的突然变化方面“训练”得更好,因此,对进料方式的变化更具“抗冲击性”。这清楚地表明了酵母之间细胞内代谢通量控制的主要差异。这些发现与工业面包酵母生产相关,在工业面包酵母生产中,一到几分钟的反应器混合时间并不罕见。观察到的热量产生以及溶解氧浓度似乎对培养物中的实际变化给出了最快的响应。建议热量测量可以成为监测和控制实验室和工业发酵罐操作中酿酒酵母生长的非常有用的工具。