PXD061558

PXD061558 is an original dataset announced via ProteomeXchange.

Title	Discovery and Characterization of a Novel Carbohydrate-Active Enzyme from the Anaerobic Fungus Piromyces finnis
Description	Dwelling in the gut of herbivores, anaerobic gut fungi (AGF) have evolved several strategies to efficiently degrade unpretreated biomass. Their genomes encode a diverse array of carbohydrate-active enzymes (CAZymes), yet exceedingly few of these enzymes have been validated. Here, we developed a predictive bioinformatic pipeline to both annotate novel putative CAZymes and validate their activity through heterologous expression. 173 fungal proteins from Piromyces finnis associated with biomass degradation were synthesized and expressed in E. coli, and 9.8% were soluble with expression levels exceeding 5% of total proteome. Among the 17 heterologously expressed proteins, a combination of sequence and structure-based homology annotation identified many multifunctional proteins having catalytic domains, such as glycoside hydrolase (GH) and carbohydrate esterase (CE), fused with repetitive dockerins. Screening across three cellulose and hemicellulose substates experimentally verified six predicted functions. One promising enzyme, celsome_012, exhibited robust and optimized activity against beechwood xylan at 37°C and pH 6.4, producing five-times more products than other recombinant proteins screened here. Overall, this study represents the first large-scale screening campaign of putative AGF CAZymes, highlighting proteins amenable to E. coli overexpression, integrating advanced sequence and structural annotation, and identifying a robust, novel fungal xylanase for detailed biochemical characterization.
HostingRepository	PRIDE
AnnounceDate	2025-07-31
AnnouncementXML	Submission_2025-07-30_21:57:21.075.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Christopher Petzold
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-06 19:25:22	ID requested
⏵ 1	2025-07-30 21:57:21	announced

Dataset with its publication pending

submitter keyword: CAZymes, JBEI, Proteomics, high-throughput screen,Heterologous expression

Christopher J. Petzold
contact affiliation	Staff Scientist Biological Systems & Engineering Division Lawrence Berkeley National Laboratory Berkeley CA 94720
contact email	cjpetzold@lbl.gov
lab head
Christopher Petzold
contact affiliation	Lawrence Berkeley National Laboratory
contact email	cjpetzold@lbl.gov
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD061558
     1. Label: PRIDE project
     2. Name: Discovery and Characterization of a Novel Carbohydrate-Active Enzyme from the Anaerobic Fungus Piromyces finnis