Engineered Fungus <i>Thermothelomyces thermophilus</i> Producing Plant Storage Proteins
2022
Larissa Balabanova | Aleksandra Seitkalieva | Yulia Yugay | Tatiana Rusapetova | Lubov Slepchenko | Anna Podvolotskaya | Margarita Yatsunskaya | Elena Vasyutkina | Oksana Son | Liudmila Tekutyeva | Yury Shkryl
An efficient <i>Agrobacterium</i>-mediated genetic transformation based on the plant binary vector pPZP-RCS2 was carried out for the multiple heterologous protein production in filamentous fungus <i>Thermothelomyces</i> <i>thermophilus</i> F-859 (formerly <i>Myceliophthora</i> <i>thermophila </i>F-859). The engineered fungus <i>Th.</i> <i>thermophilus</i> was able to produce plant storage proteins of <i>Zea</i> <i>mays</i> (α-zein Z19) and <i>Amaranthus</i> <i>hypochondriacus </i>(albumin A1) to enrich fungal biomass by valuable nutritional proteins and improved amino acid content. The mRNA levels of <i>z19</i> and <i>a1</i> genes were significantly dependent on their driving promoters: the promoter of tryptophan synthase (P<i>trpC</i>) was more efficient to express <i>a1</i>, while the promoter of translation elongation factor (P<i>tef</i>) provided much higher levels of <i>z19 </i>transcript abundance. In general, the total recombinant proteins and amino acid contents were higher in the P<i>tef</i>-containing clones. This work describes a new strategy to improve mycoprotein nutritive value by overexpression of plant storage proteins.
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