Offline optimization of trehalose accumulation in yeast Saccharomyces cerevisiae fed-batch culture

Trehalose accumulation in Saccharomyces cerevisiae cells: experimental data and structured modeling

An industrial strain of Saccharomyces cerevisiae (DGI 342) was cultivated in fed-batch cultivations at a specific growth rate of 0.2 h(-1). The yeast was then exposed to carbon or nitrogen starvation for up to 8 h, to study the effect of starvation on fermentative capacity and content of protein, trehalose and glycogen. Nitrogen starvation triggered the accumulation of trehalose and glycogen. After 8 h of starvation, the content of trehalose and glycogen was increased 4-fold and 2-fold, respectively. Carbon starvation resulted in a partial conversion of glycogen into trehalose. The trehalose content increased from 45 to 64 mg (g dry-weight)(-1), whereas the glycogen content in the same period was reduced from 55 to 5 mg (g dry-weight)(-1). Glycogen was consumed faster than trehalose during storage of the starved yeast for 1 month. Nitrogen starvation resulted in a decrease in the protein content of the yeast cells, and the fermentative capacity per gram dry-weight decreased by 40%. The protein content in the carbon-starved yeast increased as a result of starvation due to the fact that the content of glycogen was reduced. The fermentative capacity per gram dry-weight was, however, unaltered.

Yeast strains were isolated from dried sweet potatoes (hoshi-imo), a traditional preserved food in Japan. Dough fermentation ability, freeze tolerance, and growth rates in molasses, which are important characteristics of commercial baker's yeast, were compared between these yeast strains and a commercial yeast derivative that had typical characteristics of commercial strains. Classification tests including pulse-field gel electrophoresis and fermentation/assimilation ability of sugars showed that almost the stains isolated belonged to Saccharomyces cerevisiae. One strain, ONY1, accumulated intracellular trehalose at a higher level than commercial strain T128. Correlated with intracellular trehalose contents, the fermentation ability of high-sugar dough containing ONY1 was higher. ONY1 also showed higher freeze tolerance in both low-sugar and high-sugar doughs. The growth rate of ONY1 was significantly higher under batch and fed-batch cultivation conditions using either molasses or synthetic medium than that of strain T128. These results suggest that ONY1 has potential commercial use as baker's yeast for frozen dough and high-sugar dough.

The physiology of a commercial strain of bakers' yeast was studied in terms of the cell composition under different growth conditions and of its response to stress. The study comprised fed-batch experiments since this is the system used in bakers' yeast industry. The classical fed-batch fermentation procedure was modified in that the yeast cells were continuously grown to a steady-state at a dilution rate of 0.1/h in order to achieve more or less the same initial starting point in terms of cell composition. This steady-state culture was then switched to fed-batch concomitantly with exposure to stress. The highest amount of trehalose accumulation was achieved when nutrient depletion and heat stress were applied concomitantly. The highest amount of trehalose, 12%, was attained in cells stressed by both nitrogen depletion and heat stress. The protein content remained constant, although with some oscillations, at a value of 30% throughout this dual stress experiment.