PXD034630

PXD034630 is an original dataset announced via ProteomeXchange.

Title	Designed high-redox potential laccases exhibit high functional diversity
Description	White-rot fungi secrete an impressive repertoire of high-redox potential laccases (HRPLs) and peroxidases for efficient oxidation and utilization of lignin. Laccases are attractive enzymes for green-chemistry applications due to their broad substrate range and low environmental impact. Since expression of functional recombinant HRPLs is challenging, iterative directed evolution protocols have been applied to improve their expression, activity and stability. We implement a rational, stabilize-and-diversify strategy to two HRPLs that we could not functionally express: first, we use the PROSS stability-design algorithm to allow functional expression in yeast. Second, we use the stabilized enzymes as starting points for FuncLib active-site design to improve their activity and substrate diversity. Four of the FuncLib designed HRPLs and their PROSS progenitor exhibit substantial diversity in reactivity profiles against high-redox potential substrates, including lignin monomers. Combinations of 3-4 subtle mutations that change the polarity, solvation and sterics of the substrate-oxidation site explain the differences in reactivity profiles. These stable and versatile HRPLs are a step towards the generation of an effective lignin-degrading consortium of enzymes that can be secreted from yeast. More broadly, the stabilize-and-diversify strategy can be applied to other challenging enzyme families to expand the utility of natural enzymes.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:57:07.427.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD034630
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Meital Kupervaser
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	maXis

Revision	Datetime	Status	ChangeLog Entry
0	2022-06-17 08:07:47	ID requested
1	2023-03-16 12:16:21	announced
⏵ 2	2023-11-14 08:57:08	announced	2023-11-14: Updated project metadata.

10.6019/PXD034630;

submitter keyword: enzyme design, protein stability, heterologous expression, PROSS, enzyme promiscuity, FuncLib, lignin degradation,Laccase

Sarel J. Fleishman
contact affiliation	Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot 7600001, Israel
contact email	sarel@weizmann.ac.il
lab head
Meital Kupervaser
contact affiliation	Weizmann Institute of Science
contact email	meital.kupervaser@weizmann.ac.il
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD034630
     1. Label: PRIDE project
     2. Name: Designed high-redox potential laccases exhibit high functional diversity