PRODIS: Yeast surface, made different.

Each yeast cell builds its cell wall, an outer envelope that protects and shapes it. With the cell wall being the principal boundary between the cell and its environment, the cell shapes it meticulously, through intricate molecular mechanisms which are today only partially understood.

The yeast cell wall is made of an inner skeleton and an outer layer. While the inner skeleton is composed of ß-1,3- and ß-1,6-glucans, the outer layer is crafted from mannoproteins and their mannan chains. Some mannoproteins bind to glucan non-covalently, while others do so covalently, either by binding to ß-1,6-glucan through GPI anchor or to ß-1,3-glucan through characteristic Pir sequences. Such covalent binding can be used to immobilise novel, heterologous proteins to the surface of yeast cells, thus creating new biologically active materials. However, such newly-created cell surfaces sustain only low enzymatic activity.

To alleviate this shortcoming, we will modify the various molecular processes involved in the construction of the yeast cell wall, e.g. protein glycosylation and secretion, endocytosis, GPI anchor and native cell-wall protein binding. Moreover, we will expose on the yeast cell surface two biotechnologically noteworthy proteins, methionyl adenosine transferase (MAT) and sucrose phosphorylase (SPase). These two enzymes catalyse critical reactions in the production of S-adenosylmethionine and glucosylglycerol, two molecules today synthesised inside yeast cells, in the bioprocesses limited by the transport of required substrates into the cells. Thus, relocating the enzymatic reaction to the yeast surface would help the efficiency of the bioprocess.

Project leader: Prof Renata Teparić // Full Professor, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Associates:

Prof Vladimir Mrša // Full Professor, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Prof Damir Iveković // Full Professor, Laboratory for General and Inorganic Chemistry and Electroanalysis, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Prof Anita Slavica // Full Professor, Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Ksenija Lopandić, PhD // Senior Scientist, Institute of Microbiology and Microbial Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria

Branko Kozulić, PhD // CEO and R&D director of Gentius Ltd, Zadar, Croatia

Assoc Prof Igor Stuparević // Associate Professor, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Asst Prof Bojan Žunar // Assistant Professor, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Antonija Grbavac, PhD // Postdoctoral Researcher, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Mateja Lozančić, PhD // Postdoctoral Researcher, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Ana Novačić, PhD // Postdoctoral Researcher, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

mag. ing. bioproc. Egon Rešetar // Scientific Assistant, Laboratory for General and Inorganic Chemistry and Electroanalysis, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

mag. ing. biotechn. Tea Martinić Cezar // PhD Student, Laboratory for Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia

Eppendorf BioSpectrometer^® Basic

New Brunswick™ Innova^® 40

Martinić Cezar, T., Lozančić, M., Novačić, A., Matičević, A., Matijević, D., Vallée, B., Mrša, V., Teparić, R. & Žunar, B. (2023). Streamlining N-terminally anchored yeast surface display via structural insights into S. cerevisiae Pir proteins. Microbial Cell Factories, 22(1): 174. https://doi.org/10.1186/s12934-023-02183-2.

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