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ENGINEERING


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Define the aims and uncertainties

The goal of our project is to engineer yeast strains that are able to survive high radiation environments in space. Moreover, these yeast strains should be able to produce nutrients or other beneficial compounds for humans. To achieve this, yeast must synthesize a sufficient amount of melanin that will absorb space radiation to protect the cells while avoiding the potential cytotoxicity that melanin biosynthesis can cause (Giannopoulos et al., 2019). A precursor on melanin synthesis, L-DOPA, can auto-oxidate, leading to formation of free radicals that are also damaging to the cells. Another cause of L-DOPA toxicity arises from errors in translation. Due to its similarity to tyrosine, L-DOPA can be incorporated into newly synthesized proteins instead of tyrosine. Although the protective effect of melanin has been observed in several species, it is not known how the localization of melanin synthesis (intra-, extracellular, particle-limited) affects its protective functions and the potential toxicity of the synthesis pathway.

Design

There are different approaches to design a melanin production system in yeast cells. In the center of our strategy is an enzyme, tyrosinase, that mediates the synthesis of melanin precursors (L-Dopa, Dopaquinone) from amino acid tyrosine. Aromatic acids are synthesized in S. cerevisiae natively in the shikimate pathway (Fig. 1), but this yeast does not have the capacity to synthesize melanin. Hence, additional enzymes must be introduced (Fig. 2).

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Huj

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Huj

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