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. Multidimensio...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2008-07, Vol.8 (13), p.2714-2723
Main Authors: Swaim, Catherine L, Anton, Brian P, Sharma, Shamik S, Taron, Christopher H, Benner, Jack S
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Bioreactors - microbiology
Computational Biology - methods
Computer Simulation
Culture Media - chemistry
ESI-MS/MS
Fermentation
Fundamental and applied biological sciences. Psychology
Fungal Proteins - secretion
Gene Expression Regulation, Fungal
Genes, Fungal
Kluyveromyces - genetics
Kluyveromyces - metabolism
Kluyveromyces lactis
Miscellaneous
Models, Biological
Multi-dimensional
Proteins
Proteome - analysis
Proteome - secretion
Proteomics - methods
Secretome
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Summary: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.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200700764