Magnaporthe oryzae as an expression host for the production of the
unspecific peroxygenase AaeUPO from the basidiomycete Agrocybe aegerita
Abstract
The filamentous fungus Magnaporthe oryzae has the potential to be
developed as an alternative platform organism for the heterologous
production of industrially important enzymes. M. oryzae is easy to
handle, fast-growing and unlike yeast, posttranslational modifications
like N-glycosylations are similar to the human organism. Here, we
established M. oryzae as a host for the expression of the unspecific
peroxygenase from the basidiomycete Agrocybe aegerita (AaeUPO). UPOs are
attractive biocatalysts for selective oxyfunctionalization of
non-activated carbon-hydrogen bonds. To improve and simplify the
isolation of AaeUPO in M. oryzae, we fused a Magnaporthe signal peptide
for protein secretion and set it under control of the strong
EF1-promotor. The success of the heterologous production of full-length
AaeUPO in M. oryzae and the secretion of the functional enzyme was
confirmed by a peroxygenase-specific enzyme assay. These results offer
the possibility to establish the filamentous ascomycete M. oryzae as a
broad applicable alternative expression system. This is in particular
valid for proteins that cannot or not in sufficient yields produced in
established systems.