Transposons are fundamental genetic elements that have profoundly shaped the architecture of eukaryotic genomes. Yeasts and filamentous fungi have emerged as important chassis organisms for bioingredient production in synthetic biology and metabolic engineering. In this review, we summarise the current understanding and future opportunities in the development of transposon-based tools for genome engineering in these fungal systems. Fungal inverted terminal repeat (ITR) DNA transposons, as well as long terminal repeat (LTR) and non-LTR retrotransposons, can accelerate genomic mutagenesis, facilitating the screening of superior genotypes and phenotypes. CRISPR-associated transposons (CASTs) hold considerable potential for site-specific integration of large transgenes, bypassing the limitations imposed by low homologous recombination (HR) efficiency in non-Saccharomyces hosts. Overall, transposon-based tools represent a valuable and underexplored avenue to accelerate genome engineering and strain development in yeasts and filamentous fungi.
Bingyin Peng, Masahiro Tominaga, Chengqiang Wang et al.· Yeast· 1 citation
Methylotrophic yeasts are outstanding platforms for the production of recombinant proteins through precision fermentation. Growing demand for protein-based food ingredients requires economically viable secretion efficiency and space-time yields, as well as correct post-translational modifications to ensure proper food functionality. Here, we review recent advances and future potential in engineering methylotrophic yeasts for food protein production, with particular focus on Komagataella phaffii. We first summarise the production of three categories of protein food ingredients: recombinant dairy proteins, proteins for plant-based and cultivated meat applications, and sweet-tasting proteins. Addressing the dual goals of improving both yield and quality, we then review synthetic biology and metabolic engineering strategies in methylotrophic yeasts, with emphasis on genome engineering and gene expression tools, protein secretory pathway engineering, and glycoengineering. Further advances in synthetic biology, integrated with industrial process development, will be critical to unlocking future precision fermentation systems based on methylotrophic yeasts.
Bingyin Peng, Masahiro Tominaga, Jun Ishii et al.· Metabolic Engineering· 0 citations