Stephen Bell

Great work from Matthew and Daniel in @chemicalscience.rsc.org

Structural basis for substrate-dependent allostery in oxygen activation by a cytochrome P450 enzyme revealed by analysis at different temperatures Cytochrome P450 (CYP) enzymes are ubiquitous and important monooxygenases whose archetypal reaction is to insert an oxygen atom from dioxygen into unactivated carbon–hydrogen bonds. They require the orchestrated delivery of electrons as well as protons from the solvent. The latter is controlled through an “acid–alc

#allostery in a #P450 enzyme investigated by variable temperature crystallography and md simulations…. pubs.rsc.org/en/content/a...

Latest work for researchers in #CH bond activation using an engineered #P450 #enzyme #biocatalyst. Excellent work from all.

Scalable and Stereoselective C–H Bond Hydroxylation of Steroids Using a Designer Heme-Thiolate Peroxygenase Biocatalyst A challenging yet critical reaction in chemical synthesis is the selective activation of unactivated carbon–hydrogen bonds in complex molecules. Here we demonstrate the biocatalytic regio- and stereoselective hydroxylation of steroids, including progesterone, testosterone, adrenosterone, androstenedione, and dehydroepiandrosterone (DHEA), using a variant of an engineered bacterial cytochrome P450 enzyme, CYP154C35 from Nocardia otitidiscaviarum. We replaced two amino acid residues within the oxygen-binding groove of the I-helix, and this modification of the acid-alcohol pair to a glutamine and a glutamate set of residues had the effect of converting the enzyme into an efficient peroxygenase (CYP154C35QE). This enzyme performed the biocatalytic oxidation of the steroids stereoselectively to form the 16α-hydroxylated metabolites. In the presence of a substrate, CYP154C35QE was stable to relatively high concentrations of H2O2 but could function catalytically using lower concentrations. The enzyme tolerated 20% DMSO and was not denatured upon being preheated for 1 h at 40 °C. The enzyme was capable of high turnover numbers (∼6500), resulting in complete conversion of the substrate, enabling the formation, purification, and characterization of the enantiopure 16α-hydroxylated steroids at up to 120 mg scale.

Scalable and Stereoselective C–H Bond #Hydroxylation of Steroids Using a Designer #Heme-Thiolate #Peroxygenase Biocatalyst | The Journal of Organic Chemistry pubs.acs.org/doi/10.1021/...

Great work (and a lot of work) from lead author Lewis. Thanks to all for putting up with me during my stay. Thanks to the University of Adelaide for letting me go for a productive visit to explore #heme #enzyme #catalysis, #crystals and #structures.

The cytochrome P450 decarboxylase from Staphylococcus aureus can produce a diene from a C18 monounsaturated fatty acid: A spectroscopic, structural and kinetic characterisation Certain members of the bacterial cytochrome P450 152 family (CYP152) are peroxygenases that catalyse the decarboxylation of fatty acids into terminal …

I enjoyed contributing to this article from the @jarworrall.bsky.social lab at the @universityofessex.bsky.social during a visit over their summer. A first foray for me into P450 peroxygenase fatty acid decarboxylases to make alkenes from fatty acids.
www.sciencedirect.com/science/arti...

The cytochrome P450 decarboxylase from Staphylococcus aureus can produce a diene from a C18 monounsaturated fatty acid: A spectroscopic, structural and kinetic characterisation Certain members of the bacterial cytochrome P450 152 family (CYP152) are peroxygenases that catalyse the decarboxylation of fatty acids into terminal …

Our first foray into the P450 world. Thanks to @bellp450enzyme.bsky.social for sharpening our GC-MS skill set @jamesbirrell.bsky.social for the microwaves😃 and Lewis for persevering 🧪

Characterisation of a Self‐Sufficient Cytochrome P450 Enzyme From the Bacterium Thermosporothrix hazakensis and Its Conversion Into a Peroxygenase - Podgorski - 2025 - Microbial Biotechnology - Wiley Online Library enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/...

Upcycling lignin with a controlled burn A recent report by Harlington et al. introduces SyoA, a cytochrome P450 enzyme that efficiently removes a methyl group from the S-subunits of lignin. Lignin is one of the most abundant forms of renewa...

A really engaging TIBS Spotlight article highlighting our recent structures of a cytochrome P450 SyoA. Thanks to Professor Jennifer DuBois for elegantly contextualising the work within the broader field of lignin valorisation. Upcycling lignin with a controlled burn tinyurl.com/2s3w9ej6 🔥🫰

Structural insights into S-lignin O-demethylation via a rare class of heme peroxygenase enzymes - Nature Communications This article demonstrates that native P450 peroxygenases with the in-built machinery for hydrogen peroxide utilisation exist; defines the molecular template for native P450 peroxygenase activity; and ...

Primary @natcomms.nature.com article: "Structural insights into S-lignin O-demethylation via a rare class of heme peroxygenase enzymes" from ‪@bellp450enzyme.bsky.social,‬ ‪@freewhelan.bsky.social‬ and colleagues.

www.nature.com/articles/s41...

Online now - the Spotlight "Upcycling #lignin with a controlled burn" from Jennifer DuBois.

#CYPenzymes #CytochromeP450 #Biocatalysis #Bioreactor

Read it here: authors.elsevier.com/a/1lFS93S6Gf...

Upcycling lignin with a controlled burn A recent report by Harlington et al. introduces SyoA, a cytochrome P450 enzyme that efficiently removes a methyl group from the S-subunits of lignin. Lignin is one of the most abundant forms of renewa...

Upcycling lignin with a controlled burn: Trends in Biochemical Sciences www.cell.com/trends/bioch...

Characterisation of the thermophilic P450 CYP116B305 identified using metagenomics-derived sequence data from an Australian hot spring - Applied Microbiology and Biotechnology Abstract Cytochrome P450 enzymes (P450s) have gained significant attention due to their remarkable ability to oxidise unactivated C-H bonds with high regio- and stereoselectivity. Their industrial uti...

New paper investigating a new #biocatalyst from a #metagenome
Characterisation of the #thermophilic #P450 CYP116B305 identified using metagenomics-derived sequence data from an Australian hot spring. Appl Microbiol Biotechnol 109, 133 (2025). doi.org/10.1007/s002...

α-Synuclein and Dopamine Metabolites DOPAL and DOPAC: A Pathway to New Synthetic Neuromelanin Models Melanin-protein-Fe conjugates serving as new synthetic analogues of brain neuromelanin were prepared by oxidative oligomerization of dopamine (DA) and its metabolites, 3,4-dihydroxyphenyl acetaldehyde...

α-Synuclein and Dopamine Metabolites DOPAL and DOPAC: A Pathway to New Synthetic Neuromelanin Models sciltp.com/journals/bb/...
First article in the new journal Bioinorganics and Biocatalysis
@scilightpress.bsky.social

New paper in chemical science on #p450 #sterol oxidation.
@chemicalscience.rsc.org

doi.org/10.1039/D5SC...

Characterisation of the Self‐Sufficient Cytochrome P450 CYP116B234 From Rhodococcus globerulus and Its Suggested Native Role in 2‐Hydroxyphenylacetic Acid Metabolism A self-sufficient P450 gene (CYP116B234) was identified from Rhodococcus globerulus, a soil-dwelling bacterium isolated from beneath an Eucalypt tree. CYP116B234 consists of heme and reductase domain...

Characterisation of the Self‐Sufficient Cytochrome P450 CYP116B234 From Rhodococcus globerulus and Its Suggested Native Role in 2‐Hydroxyphenylacetic Acid Metabolism doi.org/10.1111/1751...

Comparing and Combining Alternative Strategies for Enhancing Cytochrome P450 Peroxygenase Activity Cytochrome P450 monooxygenase (CYP) enzymes have advantageous properties over chemical catalysts. However, it is often not feasible to use CYPs in larger-scale synthesis as they require additional cof...

Pleased to see this excellent and comprehensive piece of #enzyme #biocatalysis work from finishing PhD student Matthew Podgorski from Uni of Adelaide
Comparing and Combining Alternative Strategies for Enhancing Cytochrome P450 Peroxygenase Activity | ACS Catalysis pubs.acs.org/doi/10.1021/...

Happy to report our enzyme found a home! rdcu.be/eaGXG New kid on the block P450 O-demethylase SyoA-the specificity of a monooxygenase with the simplicity of catalysis driven by peroxide. Retrofit your P450 with this active site chemistry today! #lignin #syringol Out today in Nature Communications.

Congratulations to PhD candidate @alixharlington.bsky.social, the driving force behind this research; and thanks to @bellp450enzyme.bsky.social for an enjoyable and productive collaboration. Thanks also to Keith Shearwin for your unflagging support & Tuhin Das for your work characterising mutants.

Thanks to the #ARC and Ramsay fellowship scheme for supporting this research, and to the #AustralianSynchrotron MX beamline staff for keeping our critical infrastructure accessible and productive!

Latest work from my HDR students at UoA. A nice study on a self-sufficient thermophilic #P450 fusion protein for selective aromatic #CHbond activation. Thanks to James De Voss (#UQ) and Fiona Whelan (UoA) for their help.

Characterisation of the Cytochrome P450 Monooxygenase CYP116B46 from Tepidiphilus thermophilus as a Homogentisic Acid Generating Enzyme and its Conversion to a Peroxygenase - Akter - ChemBioChem - Wiley Online Library chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/...