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Directed evolution and semi-rational engineering of plant cell wall-degrading enzymes

Final Report Summary - DEGRADEPLANT (Directed evolution and semi-rational engineering of plant cell wall-degrading enzymes)

The aim of this project is to investigate the role of insertions and deletions (indels) in the acquisition of new enzyme functions. More specifically, our aim is to perform laboratory-evolution experiments incorporating indels to investigate whether backbone modifications can be selected for functional effects and how these changes constrain further evolutionary optimisation. Given the lack of readily facile techniques to generate libraries of indel variants, a major aspect of this project was focused on developing new methodologies and tools for random indel mutagenesis. Besides the use of the initial models of choice (i.e. glycoside hydrolases), we also expanded this project towards the use of other (promiscuous) enzymes, including a phosphotriesterase from P. diminuta (PTE) and an arylsulfatase from P. aeruginosa (PAS).

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