van Hoek P, de Hulster E, van Dijken J P, Pronk J T
Kluyver Laboratory of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands.
Biotechnol Bioeng. 2000 Jun 5;68(5):517-23.
High-cell-density fed-batch processes for bakers' yeast production will involve a low-average-specific growth rate due to the limited oxygen-transfer capacity of industrial bioreactors. The relationship between specific growth rate and fermentative capacity was investigated in aerobic, sucrose-limited fed-batch cultures of an industrial bakers' yeast strain. Using a defined mineral medium, biomass concentrations of 130 g dry weight/L were reproducibly attained. After an initial exponential-feed phase (mu = 0.18 h(-1)), oxygen-transfer limitation necessitated a gradual decrease of the specific growth rate to ca. 0.01 h(-1). Throughout fed-batch cultivation, sugar metabolism was fully respiratory, with a biomass yield of 0.5 g biomass/g sucrose(-1). Fermentative capacity (assayed off-line as ethanol production rate under anaerobic conditions with excess glucose) showed a strong positive correlation with specific growth rate. The fermentative capacity observed at the end of the process (mu = 0.01 h(-1)) was only half that observed during the exponential-feed phase (mu = 0.18 h(-1)). During fed-batch cultivation, activities of glycolytic enzymes, pyruvate decarboxylase and alcohol dehydrogenase in cell extracts did not exhibit marked changes. This suggests that changes of fermentative capacity during fed-batch cultivation were not primarily caused by regulation of the synthesis of glycolytic enzymes.
由于工业生物反应器的氧传递能力有限,用于面包酵母生产的高细胞密度补料分批培养过程将具有较低的平均比生长速率。在一株工业面包酵母菌株的好氧、蔗糖限制补料分批培养中,研究了比生长速率与发酵能力之间的关系。使用确定的矿物培养基,可重复获得130 g干重/L的生物量浓度。在初始指数补料阶段(μ = 0.18 h⁻¹)之后,氧传递限制使得比生长速率逐渐降低至约0.01 h⁻¹。在整个补料分批培养过程中,糖代谢完全是呼吸性的,生物量产率为0.5 g生物量/g蔗糖⁻¹。发酵能力(在厌氧条件下用过量葡萄糖离线测定乙醇生产率)与比生长速率呈强正相关。在过程结束时(μ = 0.01 h⁻¹)观察到的发酵能力仅为指数补料阶段(μ = 0.18 h⁻¹)观察到的一半。在补料分批培养期间,细胞提取物中糖酵解酶、丙酮酸脱羧酶和乙醇脱氢酶的活性没有明显变化。这表明补料分批培养期间发酵能力的变化并非主要由糖酵解酶合成的调节引起。