絮凝颗粒粒度分布对自絮凝酵母SPSC01乙醇耐受能力的影响

Influence of Floc Size Distribution on the Ethanol Tolerance of a Self-flocculating Yeast Strain SPSC01

摘要:

利用激光聚焦反射式颗粒测量系统, 通过调节不同的搅拌速率, 得到了分批补料培养条件下粒度分布不同的四个絮凝酵母SPSC01颗粒群体, 进而对絮凝颗粒群体分布对乙醇耐受性进行了系统研究。经过6 h、20%乙醇的冲击, 颗粒粒度为100、200、300和400 mm的自絮凝酵母SPSC01的存活率分别为3.5%、26.7%、48.8%和37.6%。这表明不同粒度分布的絮凝颗粒群体乙醇耐受性具有明显差别, 在一定粒度范围内乙醇耐受性达到最高, 乙醇耐受性最高的酵母群体的乙醇得率系数85.5%, 比乙醇耐性最低的颗粒群体提高了7.2%。粒度为100、200和300 mm的自絮凝酵母颗粒群体总麦角固醇、游离麦角固醇及海藻糖含量与粒度大小成正相关, 但在粒度为400 mm的絮凝颗粒群体中总麦角固醇、游离麦角固醇及海藻糖含量呈下降趋势, 与其乙醇耐性低于300 mm絮凝颗粒的结果相一致。对细胞膜透性的研究表明, 颗粒粒度为300 mm的絮凝酵母颗粒细胞膜通透性(P′)最低, 分别仅为颗粒粒度为100 mm和200 mm颗粒群体的43%和52%, 表明粒度分布不同的絮凝颗粒群体乙醇耐性的差别与细胞膜透性密切相关。

关键词:自絮凝酵母颗粒, 粒度分布, 在线检测, 乙醇耐性

Abstract:

Ethanol tolerance of self-flocculating yeast SPSC01 was studied in a 3-L bioreactor under fed-batch culture. Yeast floc populations with the average sizes around 100, 200, 300, and 400 μm were obtained by adjusting the mechanical stirring rates of the fermentation system. When subjected to 20% (V/V) ethanol shock for 6 h at 30°C, the remained cell viability was 3.5%、26.7%、48.8% and 37.6% for the aforementioned four floc populations, respectively. The highest ethanol yield 85.5% was achieved for the 300 μm flocs, 7.2% higher than that of the 100 μm flocs. The amounts of trehalose and ergosterol (including free ergosterol and total ergosterol) were positively correlated with the average size distributions from 100 to 300 μm. However, in the 400 μm flocs, the content of trehalose and ergosterol decreased, which coincided with its reduced ethanol tolerance compared to that of the 300 μm flocs. Furthermore, when subjected to 15% (V/V) ethanol shock at 30°C, the equilibrium nucleotide concentration and plasma membrane permeability coefficient(P′) of the 300 μm flocs accounted for only 43% and 52% respectively of those of the 100 μm and 200 μm populations. The effect of floc size distribution on the ethanol tolerance of the self-flocculating yeast strain SPSC01 was closely related to plasma membrane permeability. An optimal floc size distribution with the highest ethanol tolerance and ethanol production level could be obtained by controlling mechanical stirring speed of the bioreactor, which provides basis for the process optimization of fuel ethanol production using this self-flocculating strain.

Key words:self-flocculating yeast, floc size distribution, on-line monitoring, ethanol tolerance