A5050541919- Enzyme Catalysis and Immobilization
- Microbial Metabolic Engineering and Bioproduction
- Surface Chemistry and Catalysis
- Supramolecular Self-Assembly in Materials
- Supramolecular Chemistry and Complexes
- Biofuel production and bioconversion
- Electrochemical sensors and biosensors
- Microbial metabolism and enzyme function
- Nitric Oxide and Endothelin Effects
- Catalysis for Biomass Conversion
- Electrocatalysts for Energy Conversion
- Pancreatic function and diabetes
- Microbial Metabolites in Food Biotechnology
- Nanoporous metals and alloys
- Alcohol Consumption and Health Effects
- Diet, Metabolism, and Disease
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Click Chemistry and Applications
- Advanced Nanomaterials in Catalysis
- biodegradable polymer synthesis and properties
- Oxidative Organic Chemistry Reactions
- Writing and Handwriting Education
- Enzyme Production and Characterization
- Biochemical and Molecular Research
- Education in Diverse Contexts
University of Groningen
2015-2024
University of Pavia
2020
Pollution, accidents, and misinformation have earned the pharmaceutical chemical industry a poor public reputation, despite their undisputable importance to society. Biotechnological advances hold promise enable future of drastically reduced environmental impact rigorously more efficient production routes at same time. This is exemplified in Baeyer–Villiger reaction, which offers simple synthetic route oxidize ketones esters, but application hampered by requirement hazardous dangerous...
NADPH-oxidases (NOXs) purposefully produce reactive-oxygen-species (ROS) and are found in most kingdoms of life. The seven human NOXs each characterized by a specific expression profile fine regulation to spatio-temporally tune ROS concentration cells tissues. One the best known roles for is host protection against pathogens but themselves important second messengers involved tissue regeneration modulation pathways that induce sustain cell proliferation. As such, attractive pharmacological...
The combination of redox enzymes for redox-neutral cascade reactions has received increasing appreciation. An example is the an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO). ADH can use NADP+ to oxidize cyclohexanol form and NADPH. Both products are then used by CHMO produce ε-caprolactone. In this study, these two redox-complementary were fused, create self-sufficient bifunctional enzyme that convert alcohols esters or lactones. Three different genes fused gene...
Enzyme instability is an important limitation for the investigation and application of enzymes. Therefore, methods to rapidly effectively improve enzyme stability are highly appealing. In this study we applied a computational method (FRESCO) guide engineering alcohol dehydrogenase. Of 177 selected mutations, 25 mutations brought about significant increase in apparent melting temperature (ΔTm ≥ +3 °C). By combining 10-fold mutant was generated with Tm 94 °C (+51 relative wild type), almost...
To expand the arsenal of industrially applicable oxidative enzymes, fusions alcohol dehydrogenases with an NADPH-oxidase were designed. Three different (LbADH, TbADH, ADHA) expressed a thermostable fusion partner (PAMO C65D) and purified. The resulting bifunctional biocatalysts retained catalytic properties individual acted essentially like oxidases: transforming alcohols to ketones by using dioxygen as mild oxidant, while merely requiring amount NADP+ . In small-scale reactions, purified...
A collection of fusion biocatalysts has been generated that can be used for self-sufficient oxygenations or ketone reductions. These were created by fusing a Baeyer-Villiger monooxygenase (cyclohexanone from Thermocrispum municipale: TmCHMO) an alcohol dehydrogenase (alcohol Lactobacillus brevis: LbADH) with three different cofactor regeneration enzymes (formate Burkholderia stabilis: BsFDH; glucose Sulfolobus tokodaii: StGDH, and phosphite Pseudomonas stutzeri: PsPTDH). Their tolerance...
With the aim of applying redox-neutral cascade reactions in organic media, fusions a type II flavin-containing monooxygenase (FMO-E) and horse liver alcohol dehydrogenase (HLADH) were designed. The enzyme orientation expression vector found to influence overall fusion activity. resulting bifunctional retained catalytic properties both individual enzymes. lyophilized cell-free extract containing was applied for convergent reaction consisting cyclobutanone butane-1,4-diol different...
Incorporating noble metals in artificial metalloenzymes (ArMs) is challenging due to the lack of suitable soft coordinating ligands among natural amino acids. We present a new class ArMs featuring genetically encoded noble‐metal‐binding site based on non‐canonical thiophenol‐based acid, 4‐mercaptophenylalanine (pSHF), incorporated transcriptional regulator LmrR via stop codon suppression. demonstrate that pSHF an excellent ligand for their low oxidation states. The corresponding gold(I)...
The catalytic repertoire of nature has been expanded over the past decades by introduction artificial metalloenzymes. These are enzymes containing a synthetic metal complex or non-native ion. However, combining noble catalysis and remains challenging due to lack suitable ligands bind these complexes. So far, metalloenzyme design mostly involves in vitro approaches ligand anchoring, like covalent modification cysteine residue via supramolecular assembly. Here, we show facile strategy anchor...
Incorporating noble metals in artificial metalloenzymes (ArMs) is challenging due to the lack of suitable soft coordinating ligands among natural amino acids. We present a new class ArMs featuring genetically encoded noble‐metal‐binding site based on non‐canonical thiophenol‐based acid, 4‐mercaptophenylalanine (pSHF), incorporated transcriptional regulator LmrR via stop codon suppression. demonstrate that pSHF an excellent ligand for their low oxidation states. The corresponding gold(I)...
In this manuscript we describe how, by taking a systems approach, complex energy landscapes of supramolecular assemblies can be navigated using stoichiometry to control, with remarkable selectivity, which assembly gets populated. The perhaps counterintuitive finding is that it not necessarily the that, in one-to-one comparison, most stable wins competition for common building blocks, even though system under thermodynamic control. Instead, an individually less may completely dominate system....
In this manuscript we describe how, by taking a systems approach, complex energy landscapes of supramolecular assemblies can be navigated using stoichiometry to control, with remarkable selectivity, which assembly gets populated. The perhaps counterintuitive finding is that it not necessarily the that, in one-to-one comparison, most stable wins competition for common building blocks, even though system under thermodynamic control. Instead, an individually less may completely dominate system....