The present invention relates to eukaryotic host cells having modified
oligosaccharides which may be modified further by heterologous expression
of a set of glycosyltransferases, sugar transporters and mannosidases to
become host-strains for the production of mammalian, e.g., human
therapeutic glycoproteins. The invention provides nucleic acid molecules
and combinatorial libraries which can be used to successfully target and
express mammalian enzymatic activities such as those involved in
glycosylation to intracellular compartments in a eukaryotic host cell.
The process provides an engineered host cell which can be used to express
and target any desirable gene(s) involved in glycosylation. Host cells
with modified oligosaccharides are created or selected. N-glycans made in
the engineered host cells have a Man.sub.5GlcNAc.sub.2 core structure
which may then be modified further by heterologous expression of one or
more enzymes, e.g., glycosyltransferases, sugar transporters and
mannosidases, to yield human-like glycoproteins. For the production of
therapeutic proteins, this method may be adapted to engineer cell lines
in which any desired glycosylation structure may be obtained.