PXD067323

PXD067323 is an original dataset announced via ProteomeXchange.

Title	Characterizing Peptidase Activity of Barley Endoprotease B in Complex Technical Matrices of the Beer Brewing Process
Description	Barley cysteine endoprotease B (HvEPB) is an important enzyme in plant biology and food production. Pilot-scale fed-batch fermentation with Komagataella phaffii in minimal FM22 medium enabled controlled recombinant HvEPB (r-HvEPB) secretion. Peptidomics across four technological brewing matrices - raw barley, malt, wort and beer - revealed hydrophobic P2 preference (V, L, Y) and heterogeneous P1 selection (T, Q, G), with certain residues disfavoured (S, A, G at P2; P, I, L at P1). Cleavage motifs shifted depending on the underlying matrix, primarily reflecting differences in substrates properties and availability. Immunogenic gluten peptides, particularly from C- and B-hordeins, were efficiently cleaved. Enzyme-linked immunosorbent assay confirmed 85-89% gluten reduction across all four matrices under controlled in vitro incubation. r-HvEPB hydrolysed brewing-relevant hordeins, α/β-amylases, while LTP I and serpin Z4 were less cleaved. Weighted motif analysis and inhibition trials underscored HvEPB's pivotal role as a broad-specificity endoprotease in malt-based systems, with r-HvEPB showing significantly higher gluten hydrolysis than endogenous malt proteases.
HostingRepository	PRIDE
AnnounceDate	2026-07-28
AnnouncementXML	Submission_2026-07-28_05:48:31.254.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Christina Ludwig
SpeciesList	scientific name: Komagataella phaffii; NCBI TaxID: NEWT:460519; scientific name: Hordeum vulgare subsp. vulgare; NCBI TaxID: NEWT:112509;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-08-13 14:29:15	ID requested
⏵ 1	2026-07-28 05:48:32	announced

10.1016/j.foodchem.2026.149314;

Kerpes R, Ludwig C, Dionisio G, Brinch-Pedersen H, Becker T, Characterising peptidase activity of barley endoprotease B in complex technical matrices of the brewing process. Food Chem, 516():149314(2026) [pubmed]

submitter keyword: beer,Endoprotease B, EPB, proteolysis, peptidases, gluten

Christina Ludwig
contact affiliation	Bavarian Center for Biomolecular Mass Spectrometry (BayBioMS) Technical University of Munich (TUM) Gregor-Mendel-Stasse 4 85354 Freising
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. PXD067323
     1. Label: PRIDE project
     2. Name: Characterizing Peptidase Activity of Barley Endoprotease B in Complex Technical Matrices of the Beer Brewing Process