Proteomics guided study of Saccharomyces cerevisiae for the development
of consolidated bioprocessing chasses
Abstract
New approaches to increase productivity and reduce costs may improve the
use of lignocellulosic biomass in biorefineries, benefiting the biobased
economy and reducing political and environmental instabilities from
fossil sources. The deletion of MNN10 and PMT5 genes,
related to N- and O-mannosylation processes, in Saccharomyces
cerevisiae BY4742 was performed by CRISPR/Cas9 to evaluate the effects
on the enzymatic activity of a heterologous laccase. Next, the mutant
with the higher laccase activity, mnn10Δ, in concert with
expression of lignocellulolytic enzymes ( GH74 endoglucanase,
GH3 β-glucosidase) integrated in the Ty1 locus was used to
create a strain for a consolidated bioprocess. Proteomic analysis showed
that several proteins related to the synthesis of the cell wall, vesicle
formation, protein trafficking, and glycosylation had changes in
expression levels after deletion of the MNN10. This mutant strain
also showed increased extracellular activities of recombinant CAZYmes,
which could be a result of changes in N- and O-glycosylation or
extracellular secretion. Active heterologous enzymes can be expressed
from genes integrated into the Ty1 locus (transposon) of such
industrial and laboratory S. cerevisiae chassis and provides a
promising approach for the development of consolidated bioprocessing.