Overview

Synthetic Biology Ph.D. Student.

Project title: Synthetic biology for green chemistry: Building in vivo enzymatic cascades using Carboxylic acid reductases (CARs).

Qualifications

The University of Exeter.
Doctor of Philosophy - Ph.D. 
Biological Sciences.
Dates attended or expected graduation: 2017 – 2021.

MPhil/Ph.D. in Synthetic biology for green chemistry: building in vivo enzymatic cascades to perform example biotransformations using carboxylic acid reductases (CARs). 

BBSRC funded studentship in partnership with GlaxoSmithKline.

The University of Portsmouth.
Masters of Research (MRes). 
Biological Sciences. 
Grade: Distinction.
Dates attended: 2016 – 2017.

Accreditation in the following modules:
Level 7 FHEQ:
Research Preparation Unit.
Research Unit.

Laboratory Project Entitled: Expression of the Dengue serotype 2 & Zika Envelope Proteins and purification of potential inhibitor compounds.

The University of Portsmouth.
Bachelor of Science (B.Sc.). 
Biochemistry.
Grade: Upper second class with honors.
Dates attended: 2013 – 2016.

Accreditation in the following modules:

Level 4 FHEQ: Experimental biology, Biodiversity & Evolution, Introduction to Cell Biology & Biochemistry, Microbiology & Molecular Biology, Perspectives in Biochemistry and, Graduate Skills Level 1.

Level 5 FHEQ: Microbiology, Cell Biology, Biochemistry, Genetic Engineering, Development: How Form & Function Changes and, Macromolecules.

Level 6 FHEQ: Gene Organization and Expression, Genomics in Molecular Medicine, Biomolecular Science, Genes & Development.

Honors Project Entitled 'The Co-Crystallization of Human Serum Amyloid P Component (SAP) with Hairpin and Self-complementary Oligonucleotides.

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Research

Research interests

- Analytical Biochemistry.

- Computational Biology.

- Drug Discovery.

- Enzymology. 

- Protein Engineering.

- Structural Determination of Biomolecules.

- Synthetic Biology.

- Tropical Diseases & their Pathology.

Research projects

Synthetic biology for green chemistry: Building in vivo enzymatic cascades using Carboxylic acid reductases (CARs).

Overview: Enzyme cascades are highly attractive biocatalytic approaches for chemical transformations. These approaches envisage using a series of enzymes to perform multiple coordinated reaction steps in a single process. This has many advantages, including removing the need for intermediate purification and often allowing side products to be recycled. Enzyme cascades have been demonstrated in vitro and in vivo. Using cascades in vivo is highly attractive as there is no need to add expensive cofactors. A particular advantage would be to perform enzyme cascades in thermophiles. These can be more easily integrated into mixed chemical-biocatalytic pipelines, and the high temperatures generally eliminate competitor microbes. A particularly attractive approach would be to implement in vivo cascades utilizing carboxylic acid reductases (CARs). These enzymes reduce carboxylic acids to aldehydes, which is chemically challenging to achieve without reducing to the alcohol, and which produces large quantities of chemical waste. CARs also act as an attractive bridge reaction, as their substrates are produced by many other steps, and their products are substrates for many following reactions. A GSK/BBSRC CASE project recently concluded that has demonstrated clearly the potential of this research. Publications describing an esterase (Sayer et al. (2016) Sci. Rep., 6, 25542) and characterizing a range of CARs (Finnigan et al. (2017) ChemCatChem, 9, 1005-17) have resulted from this, with a third describing modeling currently in revision. This project aims to further develop the proof-of-principle work performed in the first project, and exploit it for practical use by GSK.
This project aims to develop in vivo thermophilic cascades using CARs as examples for industrial use.

The project will aim to develop these by:
1. Demonstrating that CAR-based enzyme cascades are effective in E. coli. 
2. Using an enzyme cascade to convert a precursor compound to an industrially relevant alcohol using an enzyme cascade in vitro and in vivo.
3. Extending the range of reactions that can be coupled to exploit the CAR enzymes.
Pilot studies have shown that these steps are all feasible. Specifically, a CAR-based enzyme cascade has been thoroughly characterized biochemically in vitro. A robust model of this cascade has been developed.

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