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Industrial PE-2 strain of Saccharomyces cerevisiae: From alcoholic fermentation to the production of recombinant proteins

机译：酿酒酵母工业PE-2菌株：从酒精发酵到重组蛋白的生产

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摘要

Saccharomyces cerevisiae is the most important microorganism used in the ethanol fermentation process. The PE-2 strain of this yeast is widely used to produce alcohol in Brazil due to its high fermentation capacity. The aim of the present study was to develop an expression system for recombinant proteins using the industrial PE-2 strain of S. cerevisiae during the alcoholic fermentation process. The protein chosen as a model for this system was CaneCPI-1, a cysteine peptidase inhibitor. A plasmid containing the CaneCPI-1 gene was constructed and yeast cells were transformed with the pYADE4_CaneCPI-1 construct. To evaluate the effect on fermentation ability, the transformed strain was used in the fermentation process with cell recycling. During the nine-hour fermentative cycles the transformed strain did not have its viability and fermentation ability affected. In the last cycle, when the fermentation lasted longer, the protein was expressed probably at the expense of ethanol once the sugars were exhausted. The recombinant protein was expressed in yeast cells, purified and submitted to assays of activity that demonstrated its functionality. Thus, the industrial PE-2 strain of S. cerevisiae can be used as a viable system for protein expression and to produce alcohol simultaneously. The findings of the present study demonstrate the possibility of producing recombinant proteins with biotechnological applications during the ethanol fermentation process.
机译：酿酒酵母是乙醇发酵过程中最重要的微生物。该酵母的PE-2菌株由于其高发酵能力而被广泛用于在巴西生产酒精。本研究的目的是在酒精发酵过程中使用酿酒酵母的工业PE-2菌株开发重组蛋白的表达系统。选择用作该系统模型的蛋白质是CaneCPI-1（一种半胱氨酸肽酶抑制剂）。构建了包含CaneCPI-1基因的质粒，并用pYADE4_CaneCPI-1构建体转化了酵母细胞。为了评估对发酵能力的影响，将转化的菌株用于细胞回收的发酵过程中。在九小时的发酵周期中，转化的菌株没有影响其活力和发酵能力。在最后一个循环中，当发酵持续更长的时间时，一旦糖耗尽，蛋白质的表达就可能以乙醇为代价。重组蛋白在酵母细胞中表达，纯化后用于证明其功能的活性测定。因此，酿酒酵母的工业PE-2菌株可用作蛋白质表达和同时生产醇的可行系统。本研究的发现证明了在乙醇发酵过程中利用生物技术应用生产重组蛋白的可能性。

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