engleski

Brettanomyces bruxellensis: Development of genetic transformation system

In biotechnology, budding yeast Brettanomyces (Dekkera) bruxellensis plays several important roles. On the one hand, it spoils wine by synthesizing “barnyard” and “band-aid” aromas, but it also adds coveted Brett character to some specialty beers and red wines. Furthermore, it can drive bioethanol fermentations and shows potential for second-generation bioethanol production. However, this potential awaits largely untapped, in part because we are still unable to tailor its physiology to the needs of the bioprocess. To tackle this challenge, our group developed the first protocol for effi- cient genetic transformation of B. bruxellensis and we are continuing our work by building much needed genetic toolbox. Here we will discuss challenges and pitfalls encountered during the development of the transformation system, starting from the optimization of transformation method to the complex fate of the transforming DNA which integrates randomly in the genome despite extensive homologies at its ends. We will detail our attempts to evade this obstacle by introducing genes which drive efficient gene targeting in S. cerevisiae. Moreover, we will outline the path for implementing state-of-the-art techniques of genetic manipulation, such as RNA interference and CRISPR/Cas9, as well as roadblocks that could arise on the way

yeast Dekkera bruxellensis, transformation

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