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Harnessing the power of enzymatic oxygen activation

Deliverables

Generation and transfer of six SSB mutant libraries to the RWTH/TUG partners for screening

Using technology of SSB, mutant libraries will be prepared and transfered to the relevant partners.

Libraries of LPMO mutants prepared

Mutants of LPMOs will be prepared and checked for activity.

Cofactor-independent oxygenases with few new functions identified within the tautomerase superfamily

Identification and functional characterisation of new cofactor-independent oxygenases within the tautomerase superfamily, with special focus on hypothetical oxygenases for which the genomic context gives clues about their physiological function.

Flavin-dependent monooxygenases produced and purified

The work will focus on eukaryotic flavin-containing monooxygenases that are essential for the detoxification of xenobiotics. The target enzymes will be produced and purified using protein chromatography.

Identification of amino acid positions that govern selectivity of three heme-dependent monooxygenases

Investigation of three heme-dependent monooxygenases for the conversion of heterocyclic compounds. Through modelling and random mutagenesis positions will be identified and mutated to elucidate on the molecular level structure-function relationships.

Identification of substrates and inhibitors of HPP oxygenase

In-depth mechanistic and structural studies of a cofactor-independent monooxygenase from the tautomerase superfamily that catalyses the oxidative cleavage of HPP.

Libraries of enzyme mutants prepared

Generate knowledge-based focussed enzyme mutant libraries

PQS-cleaving cofactor-independent dioxygenases purified and functionally characterised

Isolation of new cofactor-independent dioxygenases from the alpha/beta hydrolase fold superfamily active toward the Pseudomonas quinolone signal (PQS), exploiting natural genetic diversity; and functional characterisation of PQS-cleaving enzymes, with a focus on substrate specificity and affinity, catalytic efficiency, and applicability to interfere with quorum sensing.

OXYTRAIN vacancies advertised

For identifying ESR candidates, OXYTRAIN vacancies advertised.

Project Website

An OXYTRAIN website will be built and launched

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Publications

Characterization of a thermostable flavin-containing monooxygenase from Nitrincola lacisaponensis (NiFMO)

Author(s): Nikola Lončar, Filippo Fiorentini, Gautier Bailleul, Simone Savino, Elvira Romero, Andrea Mattevi, Marco W. Fraaije
Published in: Applied Microbiology and Biotechnology, Issue 103/4, 2019, Page(s) 1755-1764, ISSN 0175-7598
DOI: 10.1007/s00253-018-09579-w