Computational enzyme engineering for biomass breakdown: unlocking Sustainable lignocellulosic resource valorization

Abstract

Enzyme engineering is at the forefront of transformative initiatives led by the Research Hub for Biotechnology in the Built Environment, with the aim of revolutionising construction through the development of living buildings and waste valorisation. This research focuses on utilising lignocellulosic biomass, with a particular emphasis on optimising lignin degradation through enzymatic processes. Central to this work are metalloenzymes and the application of computational tools. A key focus is on Cytochrome P450 and Dye Peroxidases, with structural analyses, molecular dynamics simulations, and QM/MM methods providing deep insights into their catalytic mechanisms. Notable findings include the identification of single-site mutations in GcoAP450, such as F169A, which acts as a regulatory point for water flow, thereby enhancing catalytic efficiency. Additionally, the T296S mutation in GcoAP450 expands the enzyme’s substrate range, particularly improving lignin depolymerisation and monomer functionalisation. Structural insights into AgcAP450 and GcoAP450 highlight their potential in designing versatile P450 enzymes and guide the development of potential variants for enhanced aryl-O-demethylation, subject to further experimental validation. Studies on C4HP450 elucidate the mechanisms of aromatic hydroxylation, while research on CYP153AM.aq reveals the critical role of mutated residues in substrate specificity and catalytic efficiency. The preliminary success of the DYP-cloning project points to promising avenues for optimisation, especially in enhancing enzymatic activity and kinetics for effective lignin degradation.
In conclusion, these studies mark a significant advancement in developing more sustainable methods for addressing lignin degradation and improving biomass conversion for biofuel production. By aligning with the 2030 Sustainable Development Goals, this research supports the broader agenda of environmental sustainability, showcasing a commitment to advancing green technologies and fostering innovative solutions for a more sustainable future.

Date of Award	24 Oct 2024
Original language	English
Awarding Institution	Northumbria University
Supervisor	Warispreet Singh (Supervisor), Gary Black (Supervisor), Paul James (Supervisor) & Shafeer Kalathil (Supervisor)