About
32
Publications
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Introduction
As a synthetic biologist I am a biochemist with an engineering mindset. Particularly I am interested in the integration of computational tools and techniques into the design build test cycle of engineering biology. I am keen to further explore the use of computer aided design for novel pathways or organisms, including bioinformatics, automation, DoE and mathematical modelling.
Current institution
Additional affiliations
October 2016 - March 2019
September 2012 - present
Position
- PhD Research Student
Description
- The use of thermophiles as cell factories offers attractive opportunities for whole cell biocatalysis at elevated temperatures. Thermus thermophilus is investigated as a host organism for an enzymatic cascade reaction reducing an ester to an alcohol.
July 2010 - August 2012
BioServUK
Position
- Process Scientist
Description
- I was been responsible for many of the bulk production contracts that the company has received, including both the upstream and downstream processes to an ISO9001 standard.
Education
September 2007 - June 2010
Publications
Publications (32)
Nucleosides functionalized at the 2′-position play a crucial role in therapeutics, serving as both small-molecule drugs and modifications in therapeutic oligonucleotides. However, the synthesis of these molecules often presents substantial synthetic challenges. Here we present an approach to the synthesis of 2′-functionalized nucleosides based on e...
We report the development of an engineered aminotransferase for the synthesis of a key chiral intermediate of the anti-HIV drug Lenacapavir. Due to the sterically demanding nature of the ketone substrate, a substrate walking approach was adopted during directed evolution to unlock desired aminotransferase activity starting from a parent template (T...
Nucleosides functionalized at the 2′-position play a crucial role in therapeutics, serving as both small molecule drugs and modifications in therapeutic oligonucleotides. However, the synthesis of these molecules often presents significant synthetic challenges. In this study, we present an approach to the synthesis of 2′-functionalized nucleosides...
Selective acylation of the N-terminus over side-chains in peptides and proteins is a highly desirable but challenging reaction in chemical biology. Here we report a biomimetic approach using enzymatic in situ activation of carboxylic acids with ATP to generate reactive acyl-adenosine phosphates, which display high selectivity for the N-termini of p...
Biocatalysis is important in the discovery, development, and manufacture of pharmaceuticals. However, the identification of enzymes for target transformations of interest requires major screening efforts. Here, we report a structure-based computational workflow to prioritize protein sequences by a score based on predicted activities on substrates,...
Despite the increasing use of biocatalysis for organic synthesis, there are currently no databases that adequately capture synthetic biotransformations. The lack of a biocatalysis database prevents accelerating biocatalyst characterization efforts from being leveraged to quickly identify candidate enzymes for reactions or cascades, slowing their de...
Amide bonds occur in over half of target compounds in medicinal chemistry patents, making amide bond formation the most commonly performed reaction in the pharmaceutical industry. The growing demand for...
Biocatalysis is emerging as an attractive option for manufacturing pharmaceuticals. However, the identification of enzymes for target transformations of interest requires major screening efforts. Here we report a structure-based computational workflow to prioritize protein sequences by a score based on predicted activities on substrates, thereby re...
Biocatalysis is emerging as an attractive option for manufacturing pharmaceuticals. However, the identification of enzymes for target transformations of interest requires major screening efforts. Here we report a structure-based computational workflow to prioritize protein sequences by a score based on predicted activities on substrates, thereby re...
N‐alkanoyl‐N‐methylglucamides (MEGAs) are non‐toxic surfactants widely used as commercial ingredients, but more sustainable syntheses towards these compounds are highly desirable. Here, we present a biocatalytic route towards MEGAs and analogues using a truncated carboxylic acid reductase construct tailored for amide bond formation (CARmm‐A). CARmm...
N‐alkanoyl‐N‐methylglucamides (MEGAs) are non‐toxic surfactants widely used as commercial ingredients, but more sustainable syntheses towards these compounds are highly desirable. Here, we present a biocatalytic route towards MEGAs and analogues using a truncated carboxylic acid reductase construct tailored for amide bond formation (CARmm‐A). CARmm...
Biocatalysis is increasingly the method choice for reactions which would be challenging or environmentally unfriendly to perform via more traditional chemical methods. Concurrently, digital technologies such as computer-aided synthesis planning (CASP) are increasingly deployed to accelerate research and development. Recently, CASP tools for biocata...
Osmolytes protect microbial cells against temperature, osmolarity and other stresses. The osmolyte cyclic 2,3-diphosphoglycerate, originally isolated from the thermophilic archaeon Methanothermus fervidus, naturally protects cellular proteins under extreme conditions. The biosynthetic pathway for cyclic 2,3-diphosphoglycerate has been introduced in...
Fatty amines represent an important class of commodity chemicals which have broad applicability in different industries. The synthesis of fatty amines starts from renewable sources such as vegetable oils or animal fats, but the process has multiple drawbacks that compromise the overall effectiveness and efficiency of the synthesis. Herein, we repor...
Biocatalysis has become an important aspect of modern organic synthesis, both in academia and across the chemical and pharmaceutical industries. Its success has been largely due to a rapid expansion of the range of chemical reactions accessible, made possible by advanced tools for enzyme discovery coupled with high-throughput laboratory evolution t...
Retrosynthetic analysis for the design of synthetic routes towards target molecules is well-established in organic chemistry, and has been extended to include biocatalysis in recent years. The increasing number of transformations known to be catalysed by enzymes, whilst ultimately rendering biocatalytic retrosynthesis more powerful, necessitates th...
As the enzyme toolbox for biocatalysis has expanded, so has the potential for the construction of powerful enzymatic cascades for efficient and selective synthesis of target molecules. Additionally, recent advances in computer-aided synthesis planning are revolutionizing synthesis design in both synthetic biology and organic chemistry. However, the...
Enzyme-CBM fusion constructs immobilised on wood-derived cellulose scaffolds: a sustainable approach for continuous flow biocatalysis.
Monoacylated diamines are common motifs present in pharmaceuticals, agrochemicals and natural products. However, the synthesis of these compounds often requires selective protection/deprotection steps leading to waste and poor atom economy. Here we describe the truncation of carboxylic acid reductase (CAR) yielding an adenylation domain capable of...
Protein-based adhesives could have several advantages over petroleum-derived alternatives, including substantially lower toxicity, smaller environmental footprint and renewable sourcing. Here, we report that non-covalently crosslinked bovine serum albumin (BSA) and recombinant spider silk proteins have high adhesive strength on glass (8.53 and 6.28...
Spider silk spidroins consist of long repetitive protein strands, flanked by globular terminal domains. The globular domains are often omitted in recombinant spidroins, but are thought to be essential for the spiders’ natural spinning process. Mimicking this spinning process could be an essential step towards producing strong synthetic spider silk....
As the enzyme toolbox for biocatalysis has expanded, so has the potential for the construction of powerful enzymatic cascades for efficient and selective synthesis of target molecules. Additionally, recent advances in computer-aided synthesis planning (CASP) are revolutionizing synthesis design in both synthetic biology and organic chemistry. Howev...
Enzymes are increasingly combined into multi-enzyme systems for cost and productivity benefits. Further advantages can be gained through the use of immobilized enzymes, allowing continuous biotransformations in flow. However, the optimization of such multi-enzyme systems is challenging, particularly where immobilized enzymes are used. Here, we meet...
Spider silk spidroins consist of long repetitive protein strands, flanked by globular terminal domains. The globular domains are often omitted in recombinant spidroins, but are thought to be essential for the spiders natural spinning process. Mimicking this spinning process could be an essential step towards producing strong synthetic spider silk....
Carboxylic acid reductases (CARs) are biocatalysts of industrial importance. Their properties, especially their poor stability, render them sub-optimal for use in a bioindustrial pipeline. Here, we employed ancestral sequence reconstruction (ASR) - a burgeoning engineering tool that can identify stabilizing but enzymatically neutral mutations throu...
Multi‐step enzyme reactions offer considerable cost and productivity benefits. Process models offer a route to understanding the complexity of these reactions, and allow for their optimization. Despite the increasing prevalence of multi‐step biotransformations, there are few examples of process models for enzyme reactions. From a toolbox of charact...
Synthetic biology has a huge potential to produce the next generation of advanced materials by accessing previously unreachable (bio)chemical space. In this prospective review, we take a snapshot of current activity in this rapidly developing area, focusing on prominent examples for high-performance applications such as those required for protectiv...
Multi-step enzyme reactions offer considerable cost and productivity benefits. Process models offer a route to understanding the complexity of these reactions, and allow for their optimization. Despite the increasing prevalence of multi-step biotransformations, there are few examples of process models for enzyme reactions. From a toolbox of charact...
Carboxylic acid reductase enzymes (CARs) meet the demand in synthetic chemistry for a green and regio-specific route to aldehydes from their respective carboxylic acids. However, relatively few of these enzymes have been characterized. A sequence alignment with members of the ANL superfamily of enzymes shed light on CAR functional dynamics. Using a...
A new carboxyl esterase, AF-Est2, from the hyperthermophilic archaeon Archaeoglobus fulgidus has been cloned, over-expressed in Escherichia coli and biochemically and structurally characterized. The enzyme has high activity towards short- to medium-chain p-nitrophenyl carboxylic esters with optimal activity towards the valerate ester. The AF-Est2 h...
Questions
Questions (2)
Hi,
I was wondering if anyone has experience with the ADPglo assay from promega?
Do I need a standard curve with every plate? My feeling is probably yes if I want to be accurate, but it'll obviously use a lot more reagent.
My enzyme is generating ADP from AMP so there is no ATP contamination which makes things easier.
Thanks for any advice.
Will
I'm interested in tungsten containing aldehyde (ferredoxin) oxidoreductases from the aerobic, thermophilic bacteria Thermus thermophilus. This enzyme has been isolated from a number of thermophilic anaerobic archea in the past and in every case the enzyme was shown to be oxygen labile, with purification carried out under anaerobic conditions. I'm just curious to know whether all tungsten containing proteins are oxygen-sensitive?
