Physical and computational analysis of the yeast Kluyveromyces lactis secreted proteome.
Secretion of proteins is the most common approach to protein expression in Kluyveromyces lactis. A proteomic analysis was performed on spent fermentation medium following bioreactor propagation of a wild-type industrial strain to identify proteins naturally secreted by K. lactis cells. Multidimensional separations were conducted and RP online ESI-MS/MS analysis identified 81 secreted proteins. In addition, an in silico analysis predicted 178 K. lactis proteins to be secreted via the general secretory pathway (GSP). These two datasets were compared and approximately 70% of the K. lactis proteins detected in the culture medium possessed a GSP sequence. The detected proteins included those involved with cell wall structure and synthesis, carbohydrate metabolism, and proteolysis, a result that may have significant bearing on heterologous protein expression. Additionally, both the experimental and in silico datasets were compared to similar, previously published datasets for Candida albicans. With the methodology presented here, we provide the deepest penetration into a yeast secretome yet reported.
Swaim
Catherine L
CL
New England Biolabs, Ipswich, MA 01938, USA.
Anton
Brian P
BP
Sharma
Shamik S
SS
Taron
Christopher H
CH
Benner
Jack S
JS
D019149:Bioreactors / Q000382:microbiology; D019295:Computational Biology / Q000379:methods*; D003198:Computer Simulation; D003470:Culture Media / Q000737:chemistry; D005285:Fermentation; D005656:Fungal Proteins / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D007716:Kluyveromyces / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
D003470:Culture Media; D005656:Fungal Proteins; D020543:Proteome