iiinova — where your digital self keeps evolving

Bart Brouwer @brouwerb.bsky.social · 30/04/2026

This work was performed at the @stratinghinst.bsky.social at the university of Groningen @rug.nl.

Bart Brouwer @brouwerb.bsky.social · 30/04/2026

Finally, I would like to thank Andy and Henriette for their help with crystallization experiments and teaching me the finer art of protein crystallization. And of course, thanks to Gerard for the continued support and the many great years I had as a PhD in the @roelfesgroup.bsky.social

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Bart Brouwer @brouwerb.bsky.social · 30/04/2026

Interestingly, structural analysis showed that the new scaffolds lack the characteristic open pore of LmrR, demonstrating that this feature is not a prerequisite per se for the design of PadR-based artificial enzymes and thereby expanding the scope of available protein scaffolds for their design.

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Bart Brouwer @brouwerb.bsky.social · 30/04/2026

We identified and produced six previously uncharacterized proteins from the same protein family as LmrR (the PadR family), of which two could be used as scaffolds to construct artificial Friedel-Crafts alkylases employing genetically incorporated noncanonical amino acids as catalytic residues.

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Bart Brouwer @brouwerb.bsky.social · 30/04/2026

In this work, we performed a bioinformatics study to search for homologs of the versatile LmrR protein scaffold, which harbors an open pore that has proven to be an excellent starting point for the creation of artificial enzymes.

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Bart Brouwer @brouwerb.bsky.social · 30/04/2026

Proud to share that another one of my main PhD projects has now been published! The manuscript entitled "Exploring PadR Proteins for Artificial Enzyme Design" is now available online in #ChemBioChem. Keep readin below or check out the full paper here: doi.org/10.1002/cbic... Thread ⬇️

doi.org

Exploring PadR Proteins for Artificial Enzyme Design

Inspired by the Lactococcal multidrug resistance regulator (LmrR) protein scaffold, new phenolic acid decarboxylase transcriptional regulator (PadR) proteins were identified and evaluated as scaffold...

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Reposted by Bart Brouwer

Max Fürst @maxfus.bsky.social · 20/05/2025

🚨 preprint 2️⃣ this month: our (purely experimental🧪) venture into #ChemBio We prouldy present: ADD-tagging of proteins (or "ADDing") —a super convenient enzymatic technique to install click chemistry handles on proteins. Led by superstar @wahyuwidodo.bsky.social www.biorxiv.org/content/10.1... A 🧵👇🏽

Overview of ADD-tagging including ADP-ribosyl cyclase (ADPRC)-catalysed dinucleotide substrate generation and two step chemoenzymatic labelling of target proteins with the flavin transferase ApbE, followed by click chemistry-based functional group attachment.

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Reposted by Bart Brouwer

Roelfes Group @roelfesgroup.bsky.social · 12/05/2025

📢📢📢 PhD Opportunity! Join our research group at RUG! We're seeking a motivated PhD student for a project on biocatalytic cascade processes combining natural and designer enzymes, supported by machine learning. Click below to learn more — and feel free to share! sites.google.com/rug.nl/roelf...

sites.google.com

Roelfes group - Open Positions

PhD position on Hybrid biocatalytic cascades aided by Machine learning (1.0 FTE)

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

Last but not least, the simple and straightforward genetic incorporation of aY facilitated the application of the evolved Friedel-Crafts alkylase in whole-cells!

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

X-ray crystal structures of the parent and evolved mutant (2.2 Å and 1.2 Å!) showed a significant change in the rotameric state of the aY catalytic residue, and a narrowing of the active site cavity.

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

Directed evolution gave rise to a quadruple mutant that showed increased activity and excellent enantioselectivity (up to 95% ee).

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

Through genetic incorporation of aY into LmrR, we create an artificial Friedel-Crafts alkylase that is enantiocomplementary to a previous design featuring p-aminophenylalanine as catalytic residue.

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

We demonstrate the first example of using noncanonical 3-aminotyrosine (aY) as a catalytic residue for iminium activation in a designer enzyme

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Bart Brouwer @brouwerb.bsky.social · 23/04/2025

Very proud to share that one of my main PhD projects has been published in @chemicalscience.rsc.org, and that it has been selected as a #ChemSciPicks of the week paper! Keep reading below for more details, or check out the full paper for free online: doi.org/10.1039/D5SC... Thread ⬇️

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Reposted by Bart Brouwer

Franco Della Felice @francdefel.bsky.social · 12/02/2025

Grateful to be part of this awesome project lead by @brouwerb.bsky.social. Take a look to our preprint about unlocking iminium ion catalysis when using amino-tyrosine as a genetically encoded amino acid! 🤓 #MSCA @roelfesgroup.bsky.social

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

Thank you Franco, Andy and Gerard for the great collaboration! Looking forward to seeing it published soon, keep your eyes peeled! @roelfesgroup.bsky.social @stratinghinst.bsky.social

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

Last but not least, the simple and straightforward genetic incorporation of aY facilitated the application of the evolved Friedel-Crafts alkylase in whole-cells!

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

Directed evolution gave rise to a quadruple mutant that showed increased activity and excellent enantioselectivity (up to 95% ee).

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

Through genetic incorporation of aY into LmrR, we create an artificial Friedel-Crafts alkylase that is enantiocomplementary to a previous design featuring p-aminophenylalanine as catalytic residue. doi.org/10.26434/che...

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Bart Brouwer @brouwerb.bsky.social · 11/02/2025

Check out our latest preprint, in which we demonstrate the first example of using noncanonical 3-aminotyrosine (aY) as a catalytic residue for iminium activation in a designer enzyme! Thread ⬇️

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