The Pichia pastoris heterologous gene expression system has been utili
zed to produce attractive levels of a variety of intracellular and ext
racellular proteins of interest. Recent advances in our understanding
and application of the system have improved its utility even further.
These advances include: (1) methods for the construction of P. pastori
s strains with multiple copies of AOX1-promoter-driven expression cass
ettes; (2) mixed-feed culture strategies for high foreign protein volu
metric productivity rates; (3) methods to reduce proteolysis of some p
roducts in high cell-density culture media; (4) tested procedures for
purification of secreted products; and (5) detailed information on the
structures of N-linked oligosaccharides on P pastoris secreted protei
ns. In this review, these advances along with basic features of the P.
pastoris system are described and discussed.