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Genome, secretome and glucose transport highlight unique features of the protein production host Pichia pastoris
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Genome, secretome and glucose transport highlight unique features of the protein production host Pichia pastoris
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Journal Article
Genome, secretome and glucose transport highlight unique features of the protein production host Pichia pastoris
2009
Overview
Background
Pichia pastoris
is widely used as a production platform for heterologous proteins and model organism for organelle proliferation. Without a published genome sequence available, strain and process development relied mainly on analogies to other, well studied yeasts like
Saccharomyces cerevisiae
.
Results
To investigate specific features of growth and protein secretion, we have sequenced the 9.4 Mb genome of the type strain DSMZ 70382 and analyzed the secretome and the sugar transporters. The computationally predicted secretome consists of 88 ORFs. When grown on glucose, only 20 proteins were actually secreted at detectable levels. These data highlight one major feature of
P. pastoris
, namely the low contamination of heterologous proteins with host cell protein, when applying glucose based expression systems. Putative sugar transporters were identified and compared to those of related yeast species. The genome comprises 2 homologs to
S. cerevisiae
low affinity transporters and 2 to high affinity transporters of other Crabtree negative yeasts. Contrary to other yeasts,
P. pastoris
possesses 4 H
+
/glycerol transporters.
Conclusion
This work highlights significant advantages of using the
P. pastoris
system with glucose based expression and fermentation strategies. As only few proteins and no proteases are actually secreted on glucose, it becomes evident that cell lysis is the relevant cause of proteolytic degradation of secreted proteins. The endowment with hexose transporters, dominantly of the high affinity type, limits glucose uptake rates and thus overflow metabolism as observed in
S. cerevisiae
. The presence of 4 genes for glycerol transporters explains the high specific growth rates on this substrate and underlines the suitability of a glycerol/glucose based fermentation strategy. Furthermore, we present an open access web based genome browser
http://www.pichiagenome.org
.
Publisher
BioMed Central,BioMed Central Ltd,BMC
