Discovery of Bacterial Unspecific Peroxygenases

Area of Science:

• Biochemistry and Biotechnology
• Enzymology
• Microbial Biochemistry

Background:

• Unspecific peroxygenases (UPOs) are versatile biocatalysts found in eukaryotes, crucial for oxyfunctionalization and synthesis.
• Previously, no UPO-related enzymes were identified in prokaryotic organisms, limiting their known diversity.

Purpose of the Study:

• To discover and characterize novel bacterial heme-thiolate peroxygenases.
• To investigate the structural and functional relationship between bacterial and eukaryotic UPOs.
• To assess the biocatalytic potential of newly identified bacterial UPOs.

Main Methods:

• Structure database mining to identify potential bacterial UPO candidates.
• Proteomic analysis to confirm expression in native bacterial hosts.
• Gene cloning, heterologous expression in E. coli, and biochemical assays.

Main Results:

• Discovery of a diverse family of bacterial UPOs (BUPOs) structurally homologous to fungal UPOs.
• Confirmation of BUPO expression in native and heterologous systems.
• Demonstration of BUPOs catalyzing both peroxidase and peroxygenase activities on various substrates.

Conclusions:

• Bacterial heme-thiolate peroxygenases (BUPOs) represent a previously unknown enzyme group.
• BUPOs share key structural and functional traits with eukaryotic UPOs.
• These findings expand the known repertoire of UPOs and offer new biocatalyst candidates for industrial applications.