A5049687470- Microbial Metabolic Engineering and Bioproduction
- Biochemical and biochemical processes
- Enzyme Catalysis and Immobilization
- Biochemical Acid Research Studies
- Monoclonal and Polyclonal Antibodies Research
- Metal-Catalyzed Oxygenation Mechanisms
- Innovative Microfluidic and Catalytic Techniques Innovation
- Glycosylation and Glycoproteins Research
- Catalysis for Biomass Conversion
- Vanadium and Halogenation Chemistry
- Photosynthetic Processes and Mechanisms
- Oxidative Organic Chemistry Reactions
- Plant biochemistry and biosynthesis
- Anesthesia and Pain Management
- Microbial metabolism and enzyme function
- Advanced biosensing and bioanalysis techniques
- Analytical Chemistry and Chromatography
- Microbial Natural Products and Biosynthesis
- Biofuel production and bioconversion
- Neurological diseases and metabolism
- Enzyme-mediated dye degradation
- Click Chemistry and Applications
- CRISPR and Genetic Engineering
- Viral Infectious Diseases and Gene Expression in Insects
- Chemical Synthesis and Analysis
University of Manchester
2017-2025
Czech Academy of Sciences, Institute of Biotechnology
2020-2021
The microbial production of fine chemicals provides a promising biosustainable manufacturing solution that has led to the successful growing catalog natural products and high-value chemicals. However, development at industrial levels been hindered by large resource investments required. Here we present an integrated Design-Build-Test-Learn (DBTL) pipeline for discovery optimization biosynthetic pathways, which is designed be compound agnostic automated throughout. We initially applied...
Therapeutic oligonucleotides have emerged as a powerful drug modality with the potential to treat wide range of diseases; however, rising number therapies poses manufacturing challenge. Existing synthetic methods use stepwise extension sequences immobilized on solid supports and are limited by their scalability sustainability. We report biocatalytic approach efficiently produce in single operation where polymerases endonucleases work synergy amplify complementary embedded within catalytic...
Bio-based production of industrial chemicals using synthetic biology can provide alternative green routes from renewable resources, allowing for cleaner processes. To efficiently produce on-demand through microbial strain engineering, biomanufacturing foundries have developed automated pipelines that are largely compound agnostic in their time to delivery. Here we benchmark the capabilities a pipeline enable rapid prototyping cell factories chemically diverse industrially relevant material...
Vanadium haloperoxidases (VHPOs) are unique enzymes in biology that catalyze a challenging halogen transfer reaction and convert strong aromatic C-H bond into C-X (X = Cl, Br, I) with the use of vanadium cofactor H2O2. The VHPO catalytic cycle starts conversion hydrogen peroxide halide hypohalide on vanadate cofactor, subsequently reacts substrate. However, it is unclear whether released from enzyme or otherwise trapped within structure for halogenation organic substrates. A...
The UbiD family of reversible decarboxylases act on aromatic, heteroaromatic, and unsaturated aliphatic acids utilize a prenylated flavin mononucleotide (prFMN) as cofactor, bound adjacent to conserved Glu-Arg-Glu/Asp ionic network in the enzyme's active site. It is proposed that activation requires oxidative maturation for which two distinct isomers, prFMNketimine prFMNiminium, have been observed. also has suggested only prFMNiminium form relevant catalysis, transient cycloaddition between...
Oxygenase and peroxygenase enzymes generate intermediates at their active sites which bring about the controlled functionalization of inert C-H bonds in substrates, such as enzymatic conversion methane to methanol. To be viable catalysts, however, these must also prevent oxidative damage essential site residues, can occur during both coupled uncoupled turnover. Herein, we use a combination stopped-flow spectroscopy, targeted mutagenesis, TD-DFT calculations, high-energy resolution...
The UbiD family of reversible (de)carboxylases depends on the recently discovered prenylated-FMN (prFMN) cofactor for activity. model enzyme ferulic acid decarboxylase (Fdc1) decarboxylates unsaturated aliphatic acids via a 1,3-cycloaddition process. Protein engineering has extended Fdc1 substrate range to include (hetero)aromatic acids, although catalytic rates remain poor. This raises question how efficient decarboxylation is achieved by other members. Here, we show that Pseudomonas...
Isobutene is a high value gaseous alkene used as fuel additive and chemical building block. As an alternative to fossil derived isobutene, we here develop modified mevalonate pathway for the production of isobutene from glucose in vivo. The final step consists decarboxylation 3-methylcrotonic acid, catalysed by evolved ferulic acid decarboxylase (Fdc) enzyme. Fdc belongs prFMN-dependent UbiD enzyme family that catalyses reversible (hetero)aromatic acids or acrylic with extended conjugation....
Prenylated flavin mononucleotide (prFMN) is a recently discovered modified cofactor containing an additional nonaromatic ring, connected to the N5 and C6 atoms. This underpins reversible decarboxylation catalyzed by members of widespread UbiD enzyme family produced prenyltransferase UbiX. Oxidative maturation UbiX product prFMNH2 corresponding oxidized prFMNiminium required for ferulic acid decarboxylase (Fdc1; UbiD-type enzyme) activity. However, it unclear what role Fdc1 plays in this...
Droplet microfluidics coupled with mass spectrometry is evolving for high throughput screening. A commercial solution that can readily interface to existing instrumentation would be highly desirable and allow the technology become accessible those without experience in droplet microfluidics. Here, we demonstrate a reinjection workflow infuse samples into three common spectrometers, each different electrospray ionisation (ESI) source configurations. The ease of chip-mass (MS) coupling...
The potential of antibody conjugates with high drug loading in anticancer therapy has recently been highlighted by the approval Trastuzumab deruxtecan and Sacituzumab govitecan. These biopharmaceutical approaches have spurred interest bioconjugation strategies defined degrees drug-to-antibody ratio (DAR), particular on native antibodies. Here, a glycoengineering methodology was developed to generate DAR up eight, combining highly selective enzymatic galactosylation oxidation biorthogonal...
Chinese hamster ovary (CHO) cells are the industrial workhorse for manufacturing biopharmaceuticals, including monoclonal antibodies. CHO cell line development requires a more data-driven approach accelerated identification of hyper-productive lines. Traditional methods, which rely on time-consuming hierarchical screening, often fail to elucidate underlying cellular mechanisms driving optimal bioreactor performance. Big data analytics, coupled with advancements in ‘omics’ technologies,...
Summary Particulate corticosteroids have been described to lead greater pain improvement compared with their non‐particulate counterparts when used in epidural injections. It is hypothesised that filtering may significantly impact concentration and long‐term efficacy. We investigated if passing particulate suspensions through different commonly‐used filters affects drug dosage. Two corticosteroid formulations, triamcinolone acetonide methylprednisolone acetate, were mixed at concentrations...
Oxime formation is a convenient one-step method for ligating reducing sugars to surfaces, producing mixture of closed ring α- and β-anomers along with open-chain (E)- (Z)-isomers. Here we show that despite existing as isomers, N-acetylglucosamine (GlcNAc) oximes can still be substrates β(1,4)-galactosyltransferase (β4GalT1). β4GalT1 catalysed the galactosylation GlcNAc by galactose donor (UDP-Gal) both in solution situ on surface liposomes, conversions up 60% ca. 15-20% at liposome surface....
Ambient ionization coupled to mass spectrometry has the advantages of minimal requirements for sample preparation prior analysis which renders it suitable high throughput screening. We present a protocol that permits application this method in routine biotechnology and chemical biology laboratories are using engineered enzymes produce target compounds from substrates. show how DESI-MS can be used directly analyse activity biotransformations crude cell lysate we term DiBT-MS, is 10-1000 times...
The potential of antibody conjugates with high drug loading in anticancer therapy has recently been highlighted by the approval Trastuzumab deruxtecan and Sacituzumab govitecan. These biopharmaceutical approaches have spurred interest bioconjugation strategies defined degrees antibody-to-drug (DAR) ratios, particular on native antibodies. Here we report a glycoengineering methodology to generate DAR up eight, combining highly selective enzymatic galactosylation oxidation biorthogonal tandem...