PXD059631

PXD059631 is an original dataset announced via ProteomeXchange.

Title	A cyanobacterial screening platform for Rubisco mutant variants
Description	Rubisco is the main entry point of inorganic carbon into the biosphere and a central player in the global carbon system. Its relatively low catalytic constant as well as its tendency to also accept O2 as a substrate have made it a common target of enzyme engineering. We have developed an enzyme engineering and screening platform for Rubisco using the model cyanobacterium Synechocystis sp. PCC 6803. Starting with the Form II Rubisco from Gallionella, we first show that the enzyme can replace the native Form I Rubisco in Synechocystis and that growth rates become sensitive to CO2 and O2 levels. We address the challenge of designing a zero-shot input library, without prior experimental knowledge, by coupling the phylogenetically-guided model EVmutation with “in silico evolution”. Starting with this targeted mutagenesis library, we used competitive growth coupled to deep sequencing to compare the properties of Rubisco protein variants under different cultivation conditions. We identified an amino acid exchange which increased the thermostability of Gallionella Rubisco and conveyed resilience to detrimental exchanges. The establishment of this platform is a first step towards high-throughput screening of Rubisco variants in Synechocystis and creating optimized enzyme variants to accelerate the Calvin-Benson-Bassham cycle in cyanobacteria and possibly chloroplasts. DIA proteomics was used to compare relative quantities of the native and heterologous RuBisCO, as well as the proteome at large at different gas conditions and with or without the heterologous enzyme.
HostingRepository	PRIDE
AnnounceDate	2025-08-04
AnnouncementXML	Submission_2025-08-03_16:12:17.522.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Emil Sporre
SpeciesList	scientific name: Synechocystis sp. (strain PCC 6803 / Kazusa); NCBI TaxID: 1111708;
ModificationList	acetylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-01-10 06:30:04	ID requested
⏵ 1	2025-08-03 16:12:18	announced

10.1021/acssynbio.5c00065;

Hoffmann UA, Schuppe AZ, Knave A, Sporre E, Brismar H, Englund E, Syr, é, n PO, Hudson EP, A Cyanobacterial Screening Platform for Rubisco Mutant Variants. ACS Synth Biol, 14(7):2619-2633(2025) [pubmed]

submitter keyword: prosit, DIA, carbon fixation,RuBisCO, Cyanobacteria, EncyclopeDIA, Synechocystis PCC6803

Paul Hudson
contact affiliation	Head of the Division of Systems Biology, Department of Protein Science, at the Royal Institute of Technology (KTH), Stockholm, Sweden
contact email	paul.hudson@scilifelab.se
lab head
Emil Sporre
contact affiliation	Royal Institute of Technology (KTH)
contact email	emil.sporre@scilifelab.se
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/08/PXD059631

PRIDE project URI

• PRIDE
  1. PXD059631
     1. Label: PRIDE project
     2. Name: A cyanobacterial screening platform for Rubisco mutant variants