posted on 2021-03-02, 18:36authored byZofia
D. Jarczynska, Jakob K. H. Rendsvig, Nichlas Pagels, Veronica R. Viana, Christina S. Nødvig, Ferdinand H. Kirchner, Tomas Strucko, Michael L. Nielsen, Uffe H. Mortensen
Recent sequencing of numerous fungal
species revealed large repertoires
of putative biotechnologically relevant genes and secondary metabolite
gene clusters. However, often the commercial potential of these species
is impeded by difficulties to predict host physiological and metabolic
compatibility with a given product, and lack of adequate genetic tools.
Consequently, most heterologous production is performed in standard
hosts where genetic tools and experience are in place. However, these
species may not be suitable for all products. To increase chances
of successful heterologous production, we have created a flexible
platform, DIVERSIFY, for multispecies heterologous gene expression.
This reduces the workload to construction of a single gene expression
cassette, used to transform all DIVERSIFY strains in order to identify
the optimal cell factory host. As proof of principle of the DIVERSIFY
concept, we present the first version of our platform, DIVERSIFY 1.0,
which we have successfully used for the production of three proteins
and a metabolite in four different Aspergilli species, and for the
identification of the best producer for each of the products. Moreover,
we show that DIVERSIFY 1.0 is compatible with marker-free gene targeting
induced by the CRISPR nucleases Cas9 and MAD7.