PXD062119

PXD062119 is an original dataset announced via ProteomeXchange.

Title	Higher induction temperatures and the native secretion signal peptide promote rye prolamin 75k γ-secalin production in Komagataella phaffii
Description	Background: Gluten proteins from wheat, rye, and barley play a substantial role in human nutrition. At the same time, they can trigger several different immune reactions. This, together with their influence on the quality of grain products and their emerging role as biomaterials, makes them an interesting target for further study. The proteins' propensity for aggregation challenges heterologous eukaryotic production systems. The yeast Komagataella phaffii has demonstrated excellent qualities as a production host for heterologous proteins and was therefore investigated as a platform strain. Results: A gene coding for the rye (Secale cereale) prolamin 75k γ-secalin was cloned and inserted into K. phaffii; protein expression was verified via mass spectrometry and immunoblotting and quantified via ELISA. Different parameters were investigated regarding their effect on target protein production and endoplasmic reticulum (ER) homeostasis, including the induction temperature and co- and post-translational import into the ER. At 28°C, the cells produced 1.69-fold more 75k γ-secalin than at 20°C. The introduction of the MATα-pro-region, in conjunction with either the MATα-pre- or OST1-pre-signal, led to significantly lower 75k γ-secalin accumulation, 0.20- and 0.18-fold, respectively. No mutant showed significant changes in the unfolded protein response compared to a non-producing strain. Conclusions: K. phaffii is a suitable host for prolamin production. The absence of a significant unfolded protein response during 75k γ-secalin expression indicates little challenge of ER-homeostasis by the aggregation-prone protein. It underscores K. phaffii's imminent role in protein production. The significantly decreased protein yield through the common protein secretion leader component MATα-pro demonstrates the need for further investigation into the role of secretion signals in optimizing K. phaffii as a production platform for repetitive, aggregation-prone proteins.
HostingRepository	PRIDE
AnnounceDate	2025-09-09
AnnouncementXML	Submission_2025-09-09_07:07:14.143.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Christina Ludwig
SpeciesList	scientific name: Komagataella phaffii; NCBI TaxID: 460519; scientific name: Secale cereale; NCBI TaxID: 4550;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-21 08:01:58	ID requested
⏵ 1	2025-09-09 07:07:15	announced

10.1186/s12934-025-02809-7;

B, ü, chner K, Ludwig C, Kerpes R, Becker T, -secalin production in Komagataella phaffii. Microb Cell Fact, 24(1):185(2025) [pubmed]

submitter keyword: secretion signal peptides, prolamin,Komagataella phaffii, ERAD, Secale cereale, UPR, gluten

Christina Ludwig
contact affiliation	Bavarian Center for Biomolecular Mass Spectrometry (BayBioMS) Technical University Munich Gregor-Mendel-Straße 4 85354 Freising GERMANY
contact email	tina.ludwig@tum.de
lab head
Christina Ludwig
contact affiliation	TU Munich
contact email	tina.ludwig@tum.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD062119
     1. Label: PRIDE project
     2. Name: Higher induction temperatures and the native secretion signal peptide promote rye prolamin 75k γ-secalin production in Komagataella phaffii