Transcriptional Response of Pichia pastoris in Fed-Batch Cultivations for Lipase Production
Author Information
Author(s): David Resina, Mónika Bollók, Narendar Khatri, Francisco Valero, Peter Neubauer, Pau Ferrer
Primary Institution: Departament d'Enginyeria Química, Universitat Autònoma de Barcelona
Hypothesis
Does the overexpression of Rhizopus oryzae lipase trigger the unfolded protein response (UPR) in Pichia pastoris?
Conclusion
The study found that ROL overexpression triggers the UPR in Pichia pastoris, which can create a bottleneck in the production process.
Supporting Evidence
- ROL overexpression leads to the up-regulation of UPR-related genes like KAR2 and PDI.
- Transcriptional levels of the analyzed genes were generally higher at higher growth rates.
- The bead-based sandwich hybridization assay proved reliable for quantifying specific mRNA species.
Takeaway
When yeast cells make a lot of a special protein, it can stress them out, but scientists found a way to measure how stressed they get.
Methodology
The study used a magnetic bead-based sandwich hybridization system to analyze transcriptional levels of specific genes during different cultivation conditions.
Limitations
Further improvement of cell lysis protocols and probe design for some specific genes is needed.
Digital Object Identifier (DOI)
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